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A ptychographical coherent diffractive imaging experiment in the water window with focused soft X-rays at 500 eV is reported. An X-ray beam with high degree of coherence was selected for ptychography at the P04 beamline of PETRA III synchrotron radiation source. The beam coherence was measured with the newly developed non-redundant array method, and a coherence length of 4.1 µm and global degree of coherence of 35% at 100 µm exit slit opening in the vertical direction were determined. A pinhole, 2.6 µm in size, selected the coherent part of the beam that was used to obtain ptychographic reconstruction results of a lithographically manufactured test sample and a fossil diatom. The achieved resolution was 53 nm for the test sample and was only limited by the size of the detector. The diatom was imaged at a resolution better than 90 nm.

Melanin within melanosomes exists as eumelanin or pheomelanin. Distributions of these melanins have been studied extensively within tissues, but less often within individual melanosomes. Here, we apply X-ray fluorescence analysis with synchrotron radiation to survey the nanoscale distribution of metals within purified melanosomes of mice. The study allows a discovery-based characterization of melanosomal metals, and, because Cu is specifically associated with eumelanin, a hypothesis-based test of the 'casing model' predicting that melanosomes contain a pheomelanin core surrounded by a eumelanin shell. Analysis of Cu, Ca, and Zn shows variable concentrations and distributions, with Ca/Zn highly correlated, and at least three discrete patterns for the distribution of Cu vs. Ca/Zn in different melanosomes - including one with a Cu-rich shell surrounding a Ca/Zn-rich core. Thus, the results support predictions of the casing model, but also suggest that in at least some tissues and genetic contexts, other arrangements of melanin may co-exist., (© 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Melanosomes are highly specialized organelles that produce and store the pigment melanin, thereby fulfilling essential functions within their host organism. Besides having obvious cosmetic consequences--determining the color of skin, hair and the iris--they contribute to photochemical protection from ultraviolet radiation, as well as to vision (by defining how much light enters the eye). Though melanosomes can be beneficial for health, abnormalities in their structure can lead to adverse effects. Knowledge of their ultrastructure will be crucial to gaining insight into the mechanisms that ultimately lead to melanosome-related diseases. However, due to their small size and electron-dense content, physiologically intact melanosomes are recalcitrant to study by common imaging techniques such as light and transmission electron microscopy. In contrast, X-ray-based methodologies offer both high spatial resolution and powerful penetrating capabilities, and thus are well suited to study the ultrastructure of electron-dense organelles in their natural, hydrated form. Here, we report on the application of small-angle X-ray scattering--a method effective in determining the three-dimensional structures of biomolecules--to whole, hydrated murine melanosomes. The use of complementary information from the scattering signal of a large ensemble of suspended organelles and from single, vitrified specimens revealed a melanosomal sub-structure whose surface and bulk properties differ in two commonly used inbred strains of laboratory mice. Whereas melanosomes in C57BL/6J mice have a well-defined surface and are densely packed with 40-nm units, their counterparts in DBA/2J mice feature a rough surface, are more granular and consist of 60-nm building blocks. The fact that these strains have different coat colors and distinct susceptibilities to pigment-related eye disease suggest that these differences in size and packing are of biological significance.

The aim of the experiment was to determine the impact of heat stress on nutrient digestibility and nitrogen balance in sheep fed silages differing in fibre quality. The digestibility trial was conducted at three different ambient temperatures (15°C, 25°C and 35°C for 24 h/d). The tested brown-midrib maize (Bm) silage had a higher nutrient digestibility, except for ether extract (EE) and a higher metabolisable energy (ME) content than the control maize (Con) silage. Nitrogen (N) excretion with faeces was higher but N excretion with urine was lower for sheep fed Bm silage, subsequently N balance did not differ between the two silages. Temperature had no effect on nutrient digestibility, except for crude protein (CP), but N excretion with urine was lower at elevated temperatures. A diet by temperature interaction was found for dry matter (DM) and organic matter (OM) digestibility. When the ambient temperature increased from 15°C to 25°C, the DM and OM digestibility increased in animals fed Con silage, but decreased in animals fed Bm silage. Concomitantly, ME estimated from digestible nutrients was higher for Bm than for Con at 15°C, but no differences were found at 25°C and 35°C. Effects of diet by temperature interaction, furthermore, were observed for EE and CP digestibility. Therefore, forage quality has to be considered when feeding heat-stressed animals.

The aim of the present study was to evaluate the impact of summer temperatures in a temperate climate on mid-lactation Holstein dairy cows. Therefore, a data set was examined comprising five trials with dairy cows conducted at the experimental station of the Friedrich-Loeffler-Institute in Braunschweig, Germany. The temperature-humidity index (THI) was calculated using temperature and humidity data from the barns recorded between January 2010 and July 2012. By using a generalised additive mixed model, the impact of increasing THI on dry matter intake, milk yield and milk composition was evaluated. Dry matter intake and milk yield decreased when THI rose above 60, whilst water intake increased in a linear manner beyond THI 30. Furthermore, milk protein and milk fat content decreased continuously with increasing THI. The present results revealed that heat stress exists in Lower Saxony, Germany. However, further research is necessary to describe the mode of action of heat stress. Especially, mild heat stress has to be investigated in more detail and appropriate heat stress thresholds for temperate climates have to be developed.

The aim of the present experiment was to investigate an experimental brown midrib (Bm) maize hybrid in comparison with a control (Con) non-Bm maize hybrid on ruminal and total tract digestibility, ruminal fermentation, ruminal ingesta kinetics, nitrogen (N) utilisation and microbial efficiency. A total of six ruminally and duodenally cannulated German Holstein cows were used. Animals were fed diets of either 11.5 kg dry matter (DM) of a Con or a Bm maize silage plus 4.1 kg DM of concentrate. Ruminal and total tract digestibility of organic matter, neutral detergent fibre and acid detergent fibre did not differ between hybrids. Short-chain fatty acid concentrations and pH in the rumen were not affected, but ruminal mean retention time was lower for Diet Bm (Con: 45.4 ± 2.39 h; Bm: 40.6 ± 2.39 h; least squares means ± standard error). Cows fed Diet Bm had greater efficiency of N utilisation (Con: 30.1 ± 1.37%; Bm: 33.1 ± 1.37%) and increased flow of microbial crude protein at the duodenum (MCPF) (Con: 7.0 ± 0.37 g/MJ metabolisable energy (ME); Bm: 8.1 ± 0.37 g/MJ ME). Thus, MCPF and utilisable crude protein at the duodenum (uCP) were greater for Diet Bm (MCPF - Con: 1117 ± 52.1 g/d; Bm: 1306 ± 52.1 g/d; uCP - Con: 1594 ± 57.9 g/d; Bm: 1807 ± 57.9 g/d) and ruminal N balance was lower for Diet Bm (Con: 98.7 ± 8.92 g/d; Bm: 65.6 ± 8.92 g/d). The present results show that the Bm maize hybrid might be advantageous for dairy cow nutrition with regard to N utilisation and MCPF. However, further research is necessary to draw more precise conclusions on the potential of Bm maize hybrids in general.

X-ray ptychography is a rapidly developing phase retrieval technique that combines the experimental advantages of coherent diffractive imaging with the possibility to image extended specimens. Data collection requires imaging at several scan points with high positional accuracy, which implies susceptibility to mechanical drift. This is a well-known problem in ptychographic scans, which can reduce reconstruction quality and limit the achievable resolution. Using a simple model for positional drift, we show that a set of corrected positions can be found systematically, leading to strong improvements in the reconstruction of a Siemens star dataset severely affected by drift., (Copyright © 2012 Elsevier B.V. All rights reserved.)

The unique strengths of x-ray microscopy are high penetration depth and near-edge resonances that provide chemical information. We use ptychography, a coherent diffractive imaging technique that disposes of the requirement for isolated specimens, and demonstrate resonant imaging by exploiting resonances near the oxygen K edge to differentiate between two oxygen-containing materials. To highlight a biological system where resonant ptychography might be used for chemical mapping of unsliced cells, reconstructions of freeze-dried Deinococcus radiodurans cells at an energy of 517 eV are shown.