3,661 research outputs found
Structure of catalase determined by MicroED.
MicroED is a recently developed method that uses electron diffraction for structure determination from very small three-dimensional crystals of biological material. Previously we used a series of still diffraction patterns to determine the structure of lysozyme at 2.9 Ã… resolution with MicroED (Shi et al., 2013). Here we present the structure of bovine liver catalase determined from a single crystal at 3.2 Ã… resolution by MicroED. The data were collected by continuous rotation of the sample under constant exposure and were processed and refined using standard programs for X-ray crystallography. The ability of MicroED to determine the structure of bovine liver catalase, a protein that has long resisted atomic analysis by traditional electron crystallography, demonstrates the potential of this method for structure determination
Modeling truncated pixel values of faint reflections in MicroED images.
The weak pixel counts surrounding the Bragg spots in a diffraction image are important for establishing a model of the background underneath the peak and estimating the reliability of the integrated intensities. Under certain circumstances, particularly with equipment not optimized for low-intensity measurements, these pixel values may be corrupted by corrections applied to the raw image. This can lead to truncation of low pixel counts, resulting in anomalies in the integrated Bragg intensities, such as systematically higher signal-to-noise ratios. A correction for this effect can be approximated by a three-parameter lognormal distribution fitted to the weakly positive-valued pixels at similar scattering angles. The procedure is validated by the improved refinement of an atomic model against structure factor amplitudes derived from corrected micro-electron diffraction (MicroED) images
MicroED data collection and processing.
MicroED, a method at the intersection of X-ray crystallography and electron cryo-microscopy, has rapidly progressed by exploiting advances in both fields and has already been successfully employed to determine the atomic structures of several proteins from sub-micron-sized, three-dimensional crystals. A major limiting factor in X-ray crystallography is the requirement for large and well ordered crystals. By permitting electron diffraction patterns to be collected from much smaller crystals, or even single well ordered domains of large crystals composed of several small mosaic blocks, MicroED has the potential to overcome the limiting size requirement and enable structural studies on difficult-to-crystallize samples. This communication details the steps for sample preparation, data collection and reduction necessary to obtain refined, high-resolution, three-dimensional models by MicroED, and presents some of its unique challenges
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Atomic structures of fibrillar segments of hIAPP suggest tightly mated β-sheets are important for cytotoxicity.
hIAPP fibrils are associated with Type-II Diabetes, but the link of hIAPP structure to islet cell death remains elusive. Here we observe that hIAPP fibrils are cytotoxic to cultured pancreatic β-cells, leading us to determine the structure and cytotoxicity of protein segments composing the amyloid spine of hIAPP. Using the cryoEM method MicroED, we discover that one segment, 19-29 S20G, forms pairs of β-sheets mated by a dry interface that share structural features with and are similarly cytotoxic to full-length hIAPP fibrils. In contrast, a second segment, 15-25 WT, forms non-toxic labile β-sheets. These segments possess different structures and cytotoxic effects, however, both can seed full-length hIAPP, and cause hIAPP to take on the cytotoxic and structural features of that segment. These results suggest that protein segment structures represent polymorphs of their parent protein and that segment 19-29 S20G may serve as a model for the toxic spine of hIAPP
Is it for generation me? A qualitative study exploring marketing and selling plants online to millennial-aged consumers
As online selling of products like living plants increases, it is increasingly important to understand how millennial-aged consumers perceive the purchasing experience. New-media technologies like social media, e-newsletters, and other forms of digital communication are easily adopted by millennial-aged consumers. One of these tools, 360-degree video, offers novel ways to preview products offered online and look inside local brick-and-mortar stores, which can be visited in person. Sales of horticultural goods online have been slow to be developed by industry veterans, creating ample opportunities available to new ventures. This qualitative study used a series of three focus groups to answer the research questions of RQ1: What challenges exist for garden centers attracting millennials? RQ2: What are millennials preferences for purchasing live plants online? RQ3: What aspects of digital online marketing influence millennials to make decisions? RQ4: What are millennials preferences for 360-degree video? Results of this study indicate 360-degree video is not the preferred avenue for marketing plants online to millennials, however, high-quality photos and video with educational content and the use of social media could be effective
Affinity proteomics reveals elevated muscle proteins in plasma of children with cerebral malaria
Systemic inflammation and sequestration of parasitized erythrocytes are central processes in the pathophysiology of severe Plasmodium falciparum childhood malaria. However, it is still not understood why some children are more at risks to develop malaria complications than others. To identify human proteins in plasma related to childhood malaria syndromes, multiplex antibody suspension bead arrays were employed. Out of the 1,015 proteins analyzed in plasma from more than 700 children, 41 differed between malaria infected children and community controls, whereas 13 discriminated uncomplicated malaria from severe malaria syndromes. Markers of oxidative stress were found related to severe malaria anemia while markers of endothelial activation, platelet adhesion and muscular damage were identified in relation to children with cerebral malaria. These findings suggest the presence of generalized vascular inflammation, vascular wall modulations, activation of endothelium and unbalanced glucose metabolism in severe malaria. The increased levels of specific muscle proteins in plasma implicate potential muscle damage and microvasculature lesions during the course of cerebral malaria
3D biomimetic tongue-emulating surfaces for tribological applications
Oral friction on the tongue surface plays a pivotal role in mechanics of food transport, speech, sensing and hedonic responses. The highly specialized biophysical features of human tongue such as micro-papillae-dense topology, optimum wettability and deformability present architectural challenges in designing artificial tongue surfaces, and absence of such biomimetic surface impedes fundamental understanding of tongue-food/ fluid interaction. Herein, we fabricate for the first time, a 3D soft biomimetic surface that replicates the topography and wettability of a real human tongue. The 3D-printed fabrication contains a Poisson point process-based (random) papillae distribution, and is employed to micro-mould soft silicone surfaces with wettability modifications. We demonstrate the unprecedented capability of these surfaces to replicate the theoretically-defined and simulated collision probability of papillae, and to closely resemble the tribological performances of human tongue masks. These de novo biomimetic surfaces pave the way for accurate quantification of mechanical interactions in the soft oral mucosa
Severe childhood malaria syndromes defined by plasma proteome profiles
BACKGROUND
Cerebral malaria (CM) and severe malarial anemia (SMA) are the most serious life-threatening clinical syndromes of Plasmodium falciparum infection in childhood. Therefore it is important to understand the pathology underlying the development of CM and SMA, as opposed to uncomplicated malaria (UM). Different host responses to infection are likely to be reflected in plasma proteome-patterns that associate with clinical status and therefore provide indicators of the pathogenesis of these syndromes.
METHODS AND FINDINGS
Plasma and comprehensive clinical data for discovery and validation cohorts were obtained as part of a prospective case-control study of severe childhood malaria at the main tertiary hospital of the city of Ibadan, an urban and densely populated holoendemic malaria area in Nigeria. A total of 946 children participated in this study. Plasma was subjected to high-throughput proteomic profiling. Statistical pattern-recognition methods were used to find proteome-patterns that defined disease groups. Plasma proteome-patterns accurately distinguished children with CM and with SMA from those with UM, and from healthy or severely ill malaria-negative children.
CONCLUSIONS
We report that an accurate definition of the major childhood malaria syndromes can be achieved using plasma proteome-patterns. Our proteomic data can be exploited to understand the pathogenesis of the different childhood severe malaria syndromes
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