Conservation of the Segmented Germband Stage: Robustness or Pleiotropy?

Gene expression patterns of the segment polarity genes in the extended and segmented germband stage are remarkably conserved among insects. To explain the conservation of these stages, two hypotheses have been proposed. One hypothesis states that the conservation reflects a high interactivity between modules, so that mutations would have several pleiotropic effects in other parts of the body, resulting in stabilizing selection against mutational variation. The other hypothesis states that the conservation is caused by robustness of the segment polarity network against mutational changes. When evaluating the empirical evidence for these hypotheses, we found strong support for pleiotropy and little evidence supporting robustness of the segment polarity network. This points to a key role for stabilizing selection in the conservation of these stages. Finally, we discuss the implications for robustness of organizers and long-term conservation in general

Fatigue assessment for deck plates in orthotropic bridge decks

Since the 1960s, orthotropic deck plates of highway bridges have been built with large cold-formed trapezoidal stiffeners supporting a deck plate with a thickness of approx. 12 mm. The maximum cross-beam spacing is approx. 4 m. A number of these bridge decks in The Netherlands suffer from fatigue cracks in the deck plate. First cracks have been observed after about 30 years in service. In one particular movable bridge, the cracks were found after only seven years. In many other countries, this type of crack has not yet been observed. This article provides a fatigue assessment procedure for deck plates. The procedure is calibrated with the conditions and observations in The Netherlands. It gives a fatigue life prediction and takes account of inspection results quantitatively. Although aspects such as the type and thickness of the surface finishes and the traffic load may vary between countries, the principles of the assessment procedure in this article are generally applicable and can be used to identify reasons for differences in fatigue life and to develop strategies for increasing the life

Calcium negatively regulates secretion from dense granules in Toxoplasma gondii.

Apicomplexan parasites including Toxoplasma gondii and Plasmodium spp. manufacture a complex arsenal of secreted proteins used to interact with and manipulate their host environment. These proteins are organised into three principle exocytotic compartment types according to their functions: micronemes for extracellular attachment and motility, rhoptries for host cell penetration, and dense granules for subsequent manipulation of the host intracellular environment. The order and timing of these events during the parasite's invasion cycle dictates when exocytosis from each compartment occurs. Tight control of compartment secretion is, therefore, an integral part of apicomplexan biology. Control of microneme exocytosis is best understood, where cytosolic intermediate molecular messengers cGMP and Ca2+ act as positive signals. The mechanisms for controlling secretion from rhoptries and dense granules, however, are virtually unknown. Here, we present evidence that dense granule exocytosis is negatively regulated by cytosolic Ca2+ , and we show that this Ca2+ -mediated response is contingent on the function of calcium-dependent protein kinases TgCDPK1 and TgCDPK3. Reciprocal control of micronemes and dense granules provides an elegant solution to the mutually exclusive functions of these exocytotic compartments in parasite invasion cycles and further demonstrates the central role that Ca2+ signalling plays in the invasion biology of apicomplexan parasites.Medical Research Council Australian Research Counci

Demonstration of synchronised scanning Lidar measurements of 2D velocity fields in a boundary-layer wind tunnel

This paper combines the currently relevant research methodologies of scaled wind turbine model experiments in wind tunnels with remote-sensing short-range WindScanner Lidar measurement technology. The wind tunnel of the Politecnico di Milano was equipped with three wind turbine models and two short-range WindScanner Lidars to demonstrate the benefits of synchronised scanning Lidars in such experimental surroundings for the first time. The dual- Lidar system can provide fully synchronised trajectory scans with sampling time scales ranging from seconds to minutes. First, staring mode measurements were compared to hot wire probe measurements commonly used in wind tunnels. This yielded goodness of fit coefficients of 0.969 and 0.902 for the 1 Hz averaged u- and v-components of the wind speed, respectively, validating the 2D measurement capability of the Lidar scanners. Subsequently, the measurement of wake profiles on a line as well as wake area scans were executed to illustrate the applicability of Lidar scanning to measuring small scale wind flow effects. The downsides of Lidar with respect to the hot wire probes are the larger measurement probe volume and the loss of some measurements due to moving blades. In contrast, the benefits are the high flexibility in conducting both point measurements and area scanning, and the fact that remote sensing techniques do not disturb the flow while measuring. The research campaign revealed a high potential for using short-range WindScanner Lidar for accurately measuring small scale flow structures in a wind tunnel

Improved interchangeability with different corneal specular microscopes for quantitative endothelial cell analysis

Introduction: During our clinical practice and research, we encountered an interchangeability problem when using the SP-2000P and SP-3000P TopCon corneal specular microscopes (CSMs) (TopCon Medical Systems, Tokyo, Japan) regarding the endothelial cell count (ECC). We describe a method to improve interchangeability between these CSMs. Methods: Five consecutive good-quality endothelial cell photographs were obtained in 22 eyes of 11 subjects. An ECC comparison between the two CSMs was performed after (I) gauging and calibration by the manufacturer, (II) adjustment of the magnification, (III) correction after external horizontal and vertical calibration. Results: There was a statistically significant difference between the ECC of the SP-2000P and SP-3000P at the start. The SP-2000P counted an average of 500 cells/mm2 more than the SP-3000P (p=0.00). After correction for magnification and determining a correction factor based on external calibration, the difference between the ECC of the SP-2000P and the SP-3000P was then found to be 0.4 cells/mm2 and was not statistically significant (p=0.98). Discussion: We propose a method for improving interchangeability, which involves checking magnification settings, re-checking magnification calibration with external calibration devices, and then calculating correction factors. This method can be applied to various specular or confocal microscopes and their associated endothelial cell analysis software packages to be able to keep performing precise endothelial cell counts and to enable comparison of ECCs when a CSM needs to be replaced or when results from different microscopes need to be compared

Elucidating the mitochondrial proteome of Toxoplasma gondii reveals the presence of a divergent cytochrome c oxidase

The mitochondrion of apicomplexan parasites is critical for parasite survival, although the full complement of proteins that localize to this organelle has not been defined. Here we undertake two independent approaches to elucidate the mitochondrial proteome of the apicomplexan Toxoplasma gondii. We identify approximately 400 mitochondrial proteins, many of which lack homologs in the animals that these parasites infect, and most of which are important for parasite growth. We demonstrate that one such protein, termed TgApiCox25, is an important component of the parasite cytochrome c oxidase (COX) complex. We identify numerous other apicomplexan-specific components of COX, and conclude that apicomplexan COX, and apicomplexan mitochondria more generally, differ substantially in their protein composition from the hosts they infect. Our study highlights the diversity that exists in mitochondrial proteomes across the eukaryotic domain of life, and provides a foundation for defining unique aspects of mitochondrial biology in an important phylum of parasites.This work was supported by a Discovery Grant and QEII fellowship from the Australian Research Council (ARC DP110103144) to GvD

Coronavirus disease 2019 and peripheral blood eosinophil counts: a retrospective study

Purpose Eosinopenia has been described in COVID-19. With this study, we aim to study the peripheral blood eosinophil counts in COVID-19 patients and to investigate whether there is an association between the peripheral blood eosinophil counts and disease severity of COVID-19. Methods We revised the electronical medical records of confirmed COVID-19 patients with polymerase chain reaction (PCR) assays in the Groene Hart Ziekenhuis, Gouda, The Netherlands. We divided patients in mild, moderate and severe groups based on clinical severity of COVID-19. Clinical severity was based on the therapy needed and the outcome of patients. We compared clinical characteristics, laboratory results and outcome between the three groups. Results Of the 230 patients included in this study, the mild, moderate and severe groups consisted of 16.5%, 45.7% and 37.8% of the included patients, respectively. The mean age was 68 years (IQR 57-78). 63% of patients were male. A significant decrease in the peripheral eosinophil counts was found corresponding to the increase of COVID-19 severity. In the mild, moderate and severe groups, the percentage of patients with eosinopenia was 73.7%, 86.7% and 94.3%, respectively (p value 0.002). Conclusion Eosinopenia is significantly more frequent present in patients with a severe COVID-19.Clinical epidemiolog