Using detergent to enhance detection sensitivity of African trypanosomes in human CSF and blood by Loop-Mediated Isothermal Amplification (LAMP)

<p><b>Background:</b> The loop-mediated isothermal amplification (LAMP) assay, with its advantages of simplicity, rapidity and cost effectiveness, has evolved as one of the most sensitive and specific methods for the detection of a broad range of pathogenic microorganisms including African trypanosomes. While many LAMP-based assays are sufficiently sensitive to detect DNA well below the amount present in a single parasite, the detection limit of the assay is restricted by the number of parasites present in the volume of sample assayed; i.e. 1 per µL or 103 per mL. We hypothesized that clinical sensitivities that mimic analytical limits based on parasite DNA could be approached or even obtained by simply adding detergent to the samples prior to LAMP assay.</p> <p><b>Methodology/Principal Findings:</b> For proof of principle we used two different LAMP assays capable of detecting 0.1 fg genomic DNA (0.001 parasite). The assay was tested on dilution series of intact bloodstream form Trypanosoma brucei rhodesiense in human cerebrospinal fluid (CSF) or blood with or without the addition of the detergent Triton X-100 and 60 min incubation at ambient temperature. With human CSF and in the absence of detergent, the LAMP detection limit for live intact parasites using 1 µL of CSF as the source of template was at best 103 parasites/mL. Remarkably, detergent enhanced LAMP assay reaches sensitivity about 100 to 1000-fold lower; i.e. 10 to 1 parasite/mL. Similar detergent-mediated increases in LAMP assay analytical sensitivity were also found using DNA extracted from filter paper cards containing blood pretreated with detergent before card spotting or blood samples spotted on detergent pretreated cards.</p> <p><b>Conclusions/Significance:</b> This simple procedure for the enhanced detection of live African trypanosomes in biological fluids by LAMP paves the way for the adaptation of LAMP for the economical and sensitive diagnosis of other protozoan parasites and microorganisms that cause diseases that plague the developing world.</p&gt

Serine residue 115 of MAPK-activated protein kinase MK5 is crucial for its PKA-regulated nuclear export and biological function

The mitogen-activated protein kinase-activated protein kinase-5 (MK5) resides predominantly in the nucleus of resting cells, but p38MAPK, extracellular signal-regulated kinases-3 and -4 (ERK3 and ERK4), and protein kinase A (PKA) induce nucleocytoplasmic redistribution of MK5. The mechanism by which PKA causes nuclear export remains unsolved. In the study reported here we demonstrated that Ser-115 is an in vitro PKA phosphoacceptor site, and that PKA, but not p38MAPK, ERK3 or ERK4, is unable to redistribute MK5 S115A to the cytoplasm. However, the phosphomimicking MK5 S115D mutant resides in the cytoplasm in untreated cells. While p38MAPK, ERK3 and ERK4 fail to trigger nuclear export of the kinase dead T182A and K51E MK5 mutants, S115D/T182A and K51E/S115D mutants were able to enter the cytoplasm of resting cells. Finally, we demonstrated that mutations in Ser-115 affect the biological properties of MK5. Taken together, our results suggest that Ser-115 plays an essential role in PKA-regulated nuclear export of MK5, and that it also may regulate the biological functions of MK5

A comparison of bioclimatic conditions on Franz Josef Land (the Arctic) between the turn of the nineteenth to twentieth century and present day

The paper presents the variability of meteorological conditions: air temperature, wind speed and relative air humidity; and biometeorological indices: wind chill temperature, predicted clothing insulation and accepted level of physical activity on Franz Josef Land (in Teplitz Bay and Calm Bay) in the years 1899–1931. It employs meteorological measurements taken during four scientific expeditions to the study area. The analysis mainly covered the period October–April, for which the most complete data set is available. For that period of the year, which includes the part of the year with the Franz Josef Land’s coldest air temperatures, the range and nature of changes in meteorological and biometeorological conditions between historical periods and the modern period (1981–2010) were studied. The data analysis revealed that during the three oldest expeditions (which took place in the years 1899–1914), the biometeorological conditions in the study area were more harsh to humans than in the modern period (1981–2010) or similarly harsh. In contrast, during the 1930/1931 expedition, which represents the Early Twentieth CenturyWarming (ETCW), conditions were clearly more favourable (including predicted clothing insulation being 0.3 clo lower and 4.0 °C higher wind chill temperature than conditions observed nowadays)

Loss and Damage in the Rapidly Changing Arctic

Arctic climate change is happening much faster than the global average. Arctic change also has global consequences, in addition to local ones. Scientific evidence shows that meltwater of Arctic sources contributes to sea-level rise significantly while accounting for 35% of current global sea-level rise. Arctic communities have to find ways to deal with rapidly changing environmental conditions that are leading to social impacts such as outmigration, similarly to the global South. International debates on Loss and Damage have not addressed the Arctic so far. We review literature to show what impacts of climate change are already visible in the Arctic, and present local cases in order to provide empirical evidence of losses and damages in the Arctic region. This evidence is particularly well presented in the context of outmigration and relocation of which we highlight examples. The review reveals a need for new governance mechanisms and institutional frameworks to tackle Loss and Damage. Finally, we discuss what implications Arctic losses and damages have for the international debate

Climate simulations for 1880-2003 with GISS modelE

We carry out climate simulations for 1880-2003 with GISS modelE driven by ten measured or estimated climate forcings. An ensemble of climate model runs is carried out for each forcing acting individually and for all forcing mechanisms acting together. We compare side-by-side simulated climate change for each forcing, all forcings, observations, unforced variability among model ensemble members, and, if available, observed variability. Discrepancies between observations and simulations with all forcings are due to model deficiencies, inaccurate or incomplete forcings, and imperfect observations. Although there are notable discrepancies between model and observations, the fidelity is sufficient to encourage use of the model for simulations of future climate change. By using a fixed well-documented model and accurately defining the 1880-2003 forcings, we aim to provide a benchmark against which the effect of improvements in the model, climate forcings, and observations can be tested. Principal model deficiencies include unrealistically weak tropical El Nino-like variability and a poor distribution of sea ice, with too much sea ice in the Northern Hemisphere and too little in the Southern Hemisphere. The greatest uncertainties in the forcings are the temporal and spatial variations of anthropogenic aerosols and their indirect effects on clouds.Comment: 44 pages; 19 figures; Final text accepted by Climate Dynamic