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PALM imaging Six movies of ~ 50,000 frames were acquired for each condition at 30 ms/frame. The axial drift was corrected in real time with a perfect focus system. A cylindrical lens was added to the system to induce astigmatism in the point-spread function (PSF) of the optical setup. 300,000 detections were collected on average per movie. Single molecule detection and localization was performed with a modified version of themultiple-target tracking algorithm. The 3D position of single detection was inferred from the lateral elongation of the PSF. The lateral drift of the sample was corrected by using fluorescent beads (TetraSpeck microspheres). To correct for blinking of the Dendra2 fluorophore, detections in a disk of 30 nm and adjacent in time were grouped and averaged. Nuclei and nucleoli were automatically detected and segmented for further processing.. The triangulation of the areas was performed with a custom python script and we used the ADS R package to estimate the four spatial statistics. In order to estimate the standard deviation and standard error associated with these measurements we performed a bootstrapping analysis of the data set. We randomly selected 10,000 detections from each original data set a 100 times and feed these sub sampled data set to the R script computing the spatial statistics. Single-molecule imaging (spaSPT) After overnight growth, cells were labeled with 50 nM PA-JF549 for ~15-30 min and washed twice (one wash: medium removed; PBS wash; replenished with fresh medium). At the end of the final wash, the medium was changed to phenol red-free medium keeping all other aspects of the medium the same (and adding back a-amanitin). Single-molecule imaging was performed on a custom-built Nikon TI microscope equipped with a 100x/NA 1.49 oil-immersion TIRF objective (Nikon apochromat CFI Apo TIRF 100x Oil), EM-CCD camera (Andor iXon Ultra 897; frame-transfer mode; vertical shift speed: 0.9 μs; -70°C), a perfect focusing system to correct for axial drift and motorized laser illumination (Ti-TIRF, Nikon), which allows an incident angle adjustment to achieve highly inclined and laminated optical sheet illuminatio. An incubation chamber maintained a humidified 37°C atmosphere with 5% CO2 and the objective was also heated to 37°C. Excitation was achieved using a 561 nm (1 W, Genesis Coherent) laser for PA-JF549. The excitation laser was modulated by an acousto-optic tunable filter (AA Opto-Electronic, AOTFnC-VIS-TN) and triggered with the camera TTL exposure output signal. The laser light was coupled into the microscope by an optical fiber and then reflected using a multi-band dichroic (405 nm/488 nm/561 nm/633 nm quad-band, Semrock) and then focused in the back focal plane of the objective. Fluorescence emission light was filtered using a single band-pass filter placed in front of the camera using the following filters : Semrock 593/40 nm bandpass filter. The microscope, cameras, and hardware were controlled through NIS-Elements software (Nikon). We recorded single-molecule tracking movies using our previously developed technique, stroboscopic photo-activation Single-Particle Tracking (spaSPT). Briefly, 1 ms 561 nm excitation (100% AOTF) of PA-JF549 was delivered at the beginning of the frame to minimize motion-blurring; 405 nm photo-activation pulses were delivered during the camera integration time (~447 μs) to minimize background and their intensity optimized to achieve a mean density of ~1 molecule per frame per nucleus. 30,000 frames were recorded per cell per experiment. The camera exposure time was 7 ms resulting in a frame rate of approximately 134 Hz (7 ms + ~447 μs per frame). spaSPT data was analyzed (localization and tracking) and converted into trajectories using a custom-written Matlab implementation of the MTT-algorithm and the following settings: Localization error: 10-6.25; deflation loops: 0; Blinking (frames): 1; max competitors: 3; max D (mm2/s): 20. We recorded ~9-10 cells per replicate and performed three independent replicates on three different days. Specifically, across three replicates we imaged 29 cells for Halo-RPB1-25R and obtained 448,362 trajectories with 690,682 unique displacements at a mean density of 1.2 localizations per frame. Similarly, we imaged 30 cells for Halo-RPB1-52R and obtained 324,928 trajectories with 498,503 unique displacements at a mean density of 0.9 localizations per frame. DATA FORMAT All data is available in CSV-files For the SPT analysis, the file format is readible by the web-version of Spot-On. The CSV format consists of comma-separated values and contains headers. Here the “frame” column contains the frame number in which the molecule was detected. The “t” column contains the timestamp. The “trajectory” column contains the trajectory number. For example, trajectory number 1 was only detected in frame 13 after which it disappeared. In contrast, trajectory number 4 was detected in frames 20, 21 22, 23 and 24. Finally, the “x” and “y” columns contain the x,y coordinates of the localization in units of micrometers (μm). &nbsp

Biomass from lignocellulosic energy crops can contribute to primary energy supply in the short term in heat and electricity applications and in the longer term in transport fuel applications. This paper estimates the optimal feedstock allocation of herbaceous and woody lignocellulosic energy crops for England and Wales based on empirical productivity models. Yield maps for Miscanthus, willow and poplar, constrained by climatic, soil and land use factors, are used to estimate the potential resource. An energy crop supply-cost curve is estimated based on the resource distribution and associated production costs. The spatial resource model is then used to inform the supply of biomass to geographically distributed demand centres, with co-firing plants used as an illustration. Finally, the potential contribution of energy crops to UK primary energy and renewable energy targets is discussed.

Because of the interest in the value of medium-term weather forecasts to UK agriculture, we attempt to quantify this value for N fertiliser use with arable crops. Model systems such as SUNDIAL provide arable N fertiliser advice by modelling the N supply from soil, but poor knowledge of future weather reduces accuracy. A weather generator was used to produce sets of simulated weather of a range of accuracies and durations, for 10 regions in England and Wales. In a series of computer simulations, we tested the effect of prior knowledge of weather following the date of N fertiliser application on the efficiency of use of applications using SUNDIAL. The changes in N leaching, denitrification and crop N uptake due to the forecast quality were calculated. Yield and gross profit changes were estimated from N uptake, for the arable industry in England and Wales. Changes in losses were small. With a perfect forecast, there was a small decrease in leaching (approximately 1 kg N ha −1 ), and still less change in denitrification. The increase in crop uptake due to a perfect 27-week weather forecast was 6 kg N ha −1 , and the increase in farm profit in England and Wales amounted to £68M per annum. With more accurate forecasts, the system can reduce the risk of under-application of N. A perfect 3-week forecast would increase uptake by an average of 2 kg N ha −1 , and increase profit nationally by £23M per annum but have negligible impact on losses. These improvements appear to be systematic and could be expected to be achieved with any recommendation system that makes explicit use of post-application weather.

The catchment simulated comprises 57 hectares of heavy clay soil managed as six arable fields. Mole and pipe drains carry surplus water into two ditches, one feeding into the other. Their combined flow was passed through a flume with an automatic water sampler, samples from which were analysed for nitrate. Measurements of nitrate concentration made during periods of water flow from 1990 to 1993 were simulated using a model comprising sub-models for leaching, mineralization, nitrogen uptake by crops and subsoil denitrification. The simulations were plotted against the measurements. For statistical evaluation, the correlation coefficient was used to assess the degree of association between the measurements and the simulations and the mean difference to assess the agreement. The correlation between the simulations and the measurements was significant in two of the three seasons, but the mean difference was significant in all three. However, taking all three seasons together gave a very highly significant correlation and a non-significant mean difference.

There is evidence that glutamine may act as a regulator of protein, free fatty acid, and glycogen metabolism. To test the hypothesis that glutamine may act as a physiological regulator of gluconeogenesis, we infused 16 normal postabsorptive volunteers with glutamine at a rate (11.4 micromol kg(-1) x min(-1)) estimated to approximate its appearance in plasma after a protein meal and assessed changes in production of glucose from glutamine, systemic glucose appearance and disposal, and uptake and release of glucose, glutamine, and alanine by forearm skeletal muscle. Although infusion of glutamine increased plasma glutamine concentration and turnover only threefold (from 0.63 +/- 0.03 to 1.95 +/- 0.10 mmol/l and from 5.43 +/- 0.24 to 14.85 +/- 0.66 micromol x kg(-1) x min(-1), respectively; P < 0.001), formation of glucose from glutamine increased sevenfold from 0.55 +/- 0.03 to 3.74 +/- 0.28 micromol x kg(-1) x min(-1) (P < 0.001). Formation of glucose from alanine was also stimulated (0.52 +/- 0.05 vs. 0.75 +/- 0.04 micromol x kg(-1) x min(-1); P < 0.001) in the absence of a change in plasma alanine concentration. Furthermore, glutamine infusion decreased its own de novo synthesis (4.55 +/- 0.22 vs. 2.81 +/- 0.62 micromol x kg(-1) x min(-1);P < 0.02) while increasing that of alanine (2.82 +/- 0.32 vs. 3.56 +/- 0.32 micromol x kg(-1) x min(-1); P < 0.002). Systemic glucose appearance, systemic glucose disposal, and forearm balance of glucose and alanine were not altered. Because the stimulatory effects of glutamine on gluconeogenesis occurred in the absence of changes in plasma insulin and glucagon levels, these results provide evidence that, in humans, glutamine may act both as a substrate and as a regulator of gluconeogenesis as well as a modulator of its own metabolism.

Hospital and health system executives charged with overseeing a facility design-and-build project should follow the examples of organizations that have successfully completed such projects. Here are just a few lessons learned from fruitful projects: Simplistic cost comparisons do not define value. Alternative design solutions offer a hospital the opportunity to test the value of its original design thinking. Functional designs reflect proven best practices.

Summary Realistic model representation of the dynamics of terrestrial ecosystems, including all relevant biophysical and biogeochemical interactions, is a prerequisite to accurate predictions of climate change. However, the current generation of models requires improvement. In particular, it is important to know the nitrogen status of soil for modelling global vegetation and the emission of pollutants such as nitrous oxide. For this reason, we collated several major datasets that record total nitrogen (TN) and other key properties of soils. From these data, we derived pedotransfer functions (PTFs) to predict TN of soils from other soil and site properties using stepwise multiple regressions. The most important predictor was found to be soil organic carbon (SOC). Other helpful soil and site properties for predicting TN included distance from the equator, soil texture class, soil group C:N ratio, pH, % clay and whether the soil contains more or less than 10% SOC. Several different PTFs are presented, giving the user a choice, depending on which soil and site properties are available. In all, we calculated that over 9000 observations were available, although not all datasets contained all the site properties listed above. The most useful PTF was found to be: TN = 0.0232 + 0.0250 ∗ C − 0.000534 ∗ D + 0.521 ∗ G + a(T ) ,f or C< 10%, and TN = 0.269 + 0.0449 ∗ C − 0.00575 ∗ D − 0.157 ∗ G + b(T ) ,f orC ≥ 10%, where C is % SOC, D is the degrees from the equator, G is a soil group factor defined as (C/Median C:N ratio of soil group), T is a factor based on texture classes (1, 2 or 3), depending on clay and sand content and a takes the value 0.0, 0.0228 or 0.0296 when T is 1, 2 or 3, respectively, and similarly b takes one of the values 0.0, 0.328 or 0.440, respectively. The PTFs were validated against data not used in their derivation, which were held back for this specific purpose. On the basis of the PTF above and the pre-existing digital soil map of the world, we publish a map of the global distribution of TN. It has been suggested that additional CO2 may be emitted from soils as a result of global warming, exacerbating warming still further. We calculate that the nitrous oxide formed as a result of the nitrogen that also mineralized could contribute a further 30% of additional warming.

Until recently, agricultural production was optimised almost exclusively for profit but now farming is under pressure to meet environmental targets. A method is presented and applied for optimising the sustainability of agricultural production systems in terms of both economics and the environment. Components of the agricultural production chain are analysed using environmental life-cycle assessment (LCA) and a financial value attributed to the resources consumed and burden imposed on the environment by agriculture, as well as to the products. The sum of the outputs is weighed against the inputs and the system considered sustainable if the value of the outputs exceeds those of the inputs. If this ratio is plotted against the sum of inputs for all levels of input, a diminishing returns curve should result and the optimum level of sustainability is located at the maximum of the curve. Data were taken from standard economic almanacs and from published LCA reports on the extent of consumption and environmental burdens resulting from farming in the UK. Land-use is valued using the concept of ecosystem services. Our analysis suggests that agricultural systems are sustainable at rates of production close to current levels practiced in the UK. Extensification of farming, which is thought to favour non-food ecosystem services, requires more land to produce the same amount of food. The loss of ecosystem services hitherto provided by natural land brought into production is greater than that which can be provided by land now under extensive farming. This loss of ecosystem service is large in comparison to the benefit of a reduction in emission of nutrients and pesticides. However, food production is essential, so the coupling of subsidies that represent a relatively large component of the economic output in EU farming, with measures to reduce pollution are well-aimed. Measures to ensure that as little extra land is brought into production as possible or that marginal land is allowed to revert to nature would seem to be equally well-aimed, even if this required more intensive use of productive areas. We conclude that current arable farming in the EU is sustainable with either realistic prices for products or some degree of subsidy and that productivity per unit area of land and greenhouse gas emission (subsuming primary energy consumption) are the most important pressures on the sustainability of farming.

A75-year-old woman presented with new onset of angina at rest associated with shortness of breath. The physical examination was significant for jugular venous distension and rales in both lungs. The chest radiogram showed bilateral lung congestion. She had ischemic ECG ST-segment and T-wave changes and serum creatine kinase-MB enzyme elevation consistent with a non–Q-wave myocardial infarction. Transthoracic echocardiography documented severe posterolateral and mild anteroseptal hypocontractility, with mild overall left ventricular function and mild aortic regurgitation. Because the symptoms were refractory to medical therapy, the patient was referred for cardiac catheterization. The coronary arteries appeared angiographically normal. However, …

Three methods of deriving weather for use in a nitrogen decision support system with a weekly time-step are described and evaluated. The simplest of these is simply the mean (M) of the three weather variables: Rainfall (R), evapotranspiration (ET) and temperature (T). To represent the variability of weather, many sets of generated data are needed but this is not possible with mean values. Two methods of generating rainfall are described: a fully stochastic simulator (FS) and a method based on partitioning the distribution into sections (SM). Temperature, T, and evapotranspiration ET are represented in both generators by sinusoidal functions. The amount of R is modelled as an empirical distribution, rain persistence as a Markov chain. All three means of deriving weather were compared directly with actual weather from the historical record and in use with the SUNDIAL DSS. The mean values of R, ET and T were reproduced satisfactorily (r > 0.99) by both FS and SM, but variability less accurately (r > 0.80 for the standard deviations of T, r > 0.97 for R and ET). The mean values of several components of the nitrogen cycle sim ulated with SUNDIAL were generally reproduced well for both methods of weather generation, but less accurately for mean weather. For leaching, the root mean square errors were 2.6, 2.3 and 11.4 kg/ha for FS, SM and M, respectively. The sectioning method generally gave a poor estimate of variation, which was significantly underestimated for the majority of variables, in the case of leaching by a factor of three. Where variance is important, FS is preferred; weather data generated by this method may be used with confidence for risk assessments of denitrification and crop N uptake.

The quality of test data for a dynamic system is affected by many factors, including setup of the test facility, calibration of the instrumentation and data acquisition system, data sampling rate and period, and data processing technique. This paper discusses the sources of errors typically associated with the dynamic testing of Stirling convertors. Methods for minimizing and reporting errors to ensure accurate, repeatable, test results for tests preformed by single or multiple parties are also presented.

The persistence and compliance of type 2 diabetic patients to different regimens of anti-hyperglycemic therapy were assessed retrospectively. The pharmacy claims from a pharmacy benefit management organization were analysed from the third quarter of 1996 to the fourth quarter of 1999. Of the 23 400 patients enrolled and initiating anti-diabetic therapy, 85% started treatment with monotherapy, 9.5% with insulin alone, 4.1% with polytherapy and 1.3% with insulin plus another therapy. Monotherapy patients were characterized as receiving metformin, sulfonylurea or another agent. For the 1-year follow-up period, 70.5% of the metformin patients, 75.3% of the sulfonylurea patients and 86.8% of the polytherapy patients underwent no regimen modification (except discontinuation). For the patients who had no modification of their medication regimen, persistence with sulfonylurea or metformin monotherapy was 65% greater than with polytherapy over a 1-year period. Compliance with sulfonylurea or metformin monotherapy was 45% greater than with polytherapy.

The isolation and structural elucidation of biologically active baccharinoids B1 [11a], B2 [12a], B3 [5a], and B7 [6a] are reported with crystal structure determinations of baccharinoid B7 and of the triacetate of baccharinoid B2. All four compounds are isomeric with 11a/12a and 5a/6a being epimeric at C13'.

Glycemic effect of adding 10-g carbohydrate portions of apple, banana, grapes, honeydew, orange, or strawberries to a standard meal on separate occasions was measured in 10 insulin-dependent diabetics monitored at home. The meal comprised 29% of total daily caloric intake and contained green beans, rice, turkey, and margarine (50% carbohydrate, 20% protein, and 30% fat). Blood-glucose response to meals containing grapes, honeydew, orange, or strawberries was slightly higher than meals containing apple, banana, or no fruit and the small amount of starch in apple and banana may have contributed to their lower blood-glucose response compared to the other fruits tested. Age, duration of diabetes, or insulin regimen did not correlate with subjects' responses to fruit.

1. Primary jejunal ulcer is reported quite infrequently in the medical literature. 2. A proven case of Primary jejunal ulcer of the “Simple” type with successful surgical removal is here reported. 3. There are no pathognomonic signs or symptoms of the lesion. Absorption of blood within the small intestine is often associated with disproportionately high temperature, as in the case reported. 4. The occasional occurrence of primary chronic ulcer of the jejunum must be kept in mind, when there is a clinical impression of ulcer and gastro-duodenal X-ray and gastroscopic findings are negative. 5. In case of excruciating pain and abdominal signs of ruptured viscus in the upper half of the abdomen, the occasional occurrence of ruptured primary jejunal ulcer must be remembered. 6. In cases of hemorrhage from the upper gastrointestinal tract, conservative management should be tempered with careful watchfulness and due regard for the fact that the occasional bleeding primary jejunal ulcer has an overwhelming tendency to rupture and is therefore probably best handled surgically.