Compact laboratory-based X-ray microscope enabling nondestructive 3D structure acquisition of mouse nephron with high speed and better user accessibility.

X-ray microscopes adopting computed tomography enable nondestructive 3D visualization of biological specimens at micron-level resolution without conventional 2D serial sectioning that is a destructive/laborious method and is routinely used for analyzing renal biopsy in clinical diagnosis of kidney diseases. Here we applied a compact commercial system of laboratory-based X-ray microscope to observe a resin-embedded osmium-stained 1-mm strip of a mouse kidney piece as a model of renal biopsy, toward a more efficient diagnosis of kidney diseases. A reconstructed computed tomography image from several hours of data collection using CCD detector allowed us to unambiguously segment a single nephron connected to a renal corpuscle, which was consistent with previous reports using serial sectioning. Histogram analysis on the segmented nephron confirmed that the proximal and distal tubules were distinguishable on the basis of their X-ray opacities. A 3D rendering model of the segmented nephron visualized a convoluted structure of renal tubules neighboring the renal corpuscle and a branched structure of efferent arterioles. Furthermore, another data collection using scientific complementary metal-oxide semiconductor detector with a much shorter data acquisition time of 15 min provided similar results from the same samples. These results suggest a potential application of the compact laboratory-based X-ray microscope to analyze mouse renal biopsy.

Nondestructive cellular-level 3D observation of mouse kidney using laboratory-based X-ray microscopy with paraffin-mediated contrast enhancement.

For three-dimensional observation of unstained bio-specimens using X-ray microscopy with computed tomography (CT), one main problem has been low contrast in X-ray absorption. Here we introduce paraffin-mediated contrast enhancement to visualize biopsy samples of mouse kidney using a laboratory-based X-tray microscope. Unlike conventional heavy-atom staining, paraffin-mediated contrast enhancement uses solid paraffin as a negative contrast medium to replace water in the sample. The medium replacement from water to paraffin effectively lowers the absorption of low-energy X-rays by the medium, which eventually enhances the absorption contrast between the medium and tissue. In this work, paraffin-mediated contrast enhancement with 8 keV laboratory X-rays was used to visualize cylindrical renal biopsies with diameters of about 0.5 mm. As a result, reconstructed CT images from 19.4 h of data collection achieved cellular-level resolutions in all directions, which provided 3D structures of renal corpuscles from a normal mouse and from a disease model mouse. These two structures with and without disease allowed a volumetric analysis showing substantial volume differences in glomerular subregions. Notably, this nondestructive method presents CT opacities reflecting elemental composition and density of unstained tissues, thereby allowing more unbiased interpretation on their biological structures.

Visualization and Control of Chemically Induced Crack Formation in All-Solid-State Lithium-Metal Batteries with Sulfide Electrolyte.

The application of lithium metal as a negative electrode in all-solid-state batteries shows promise for optimizing battery safety and energy density. However, further development relies on a detailed understanding of the chemo-mechanical issues at the interface between the lithium metal and solid electrolyte (SE). In this study, crack formation inside the sulfide SE (Li3PS4: LPS) layers during battery operation was visualized using in situ X-ray computed tomography (X-ray CT). Moreover, the degradation mechanism that causes short-circuiting was proposed based on a combination of the X-ray CT results and scanning electron microscopy images after short-circuiting. The primary cause of short-circuiting was a chemical reaction in which LPS was reduced at the lithium interface. The LPS expanded during decomposition, thereby forming small cracks. Lithium penetrated the small cracks to form new interfaces with fresh LPS on the interior of the LPS layers. This combination of reduction-expansion-cracking of LPS was repeated at these new interfaces. Lithium clusters eventually formed, thereby generating large cracks due to stress concentration. Lithium penetrated these large cracks easily, finally causing short-circuiting. Therefore, preventing the reduction reaction at the interface between the SE and lithium metal is effective in suppressing degradation. In fact, LPS-LiI electrolytes, which are highly stable to reduction, were demonstrated to prevent the repeated degradation mechanism. These findings will promote all-solid-state lithium-metal battery development by providing valuable insight into the design of the interface between SEs and lithium, where the selection of a suitable SE is vital.

Amyloid-ß fibrils assembled on ganglioside-enriched membranes contain both parallel ß-sheets and turns.

Some protein and peptide aggregates, such as those of amyloid-ß protein (Aß), are neurotoxic and have been implicated in several neurodegenerative diseases. Aß accumulates at nanoclusters enriched in neuronal lipids called gangliosides in the presynaptic neuronal membrane, and the resulting oligomeric and/or fibrous forms accelerate the development of Alzheimer's disease. Although the presence of Aß deposits at such nanoclusters is known, the mechanism of their assembly and the relationship between Aß secondary structure and topography are still unclear. Here, we first confirmed by atomic force microscopy that Aß40 fibrils can be obtained by incubating seed-free Aß40 monomers with a membrane composed of sphingomyelin, cholesterol, and the ganglioside GM1. Using Fourier transform infrared (FTIR) reflection-absorption spectroscopy, we then found that these lipid-associated fibrils contained parallel ß-sheets, whereas self-assembled Aß40 molecules formed antiparallel ß-sheets. We also found that the fibrils obtained at GM1-rich nanoclusters were generated from turn Aß40 Our findings indicate that Aß generally self-assembles into antiparallel ß-structures but can also form protofibrils with parallel ß-sheets by interacting with ganglioside-bound Aß. We concluded that by promoting the formation of parallel ß-sheets, highly ganglioside-enriched nanoclusters help accelerate the elongation of Aß fibrils. These results advance our understanding of ganglioside-induced Aß fibril formation in neuronal membranes and may help inform the development of additional therapies for Alzheimer's disease.

[Naive theories about the relationship between motives and behaviors].

People have naive theories about the relationship between motives and behaviors. Based on recent developments in attribution theories and negativity bias in social perception, we hypothesized that people would associate negative behaviors only with negative motives, while they would associate positive behaviors with not just positive, but also negative motives. These hypotheses were tested in three studies. In Studies 1 and 2, we found that behavioral information inconsistent with naive theories was best recalled under conditions with no cognitive load. However, this recall advantage dissipated when participants were under time pressure during the encoding of the behavioral information. In Study 3, participants were presented with positive and negative behaviors, and were asked to infer the actors' motives from these behaviors. The results showed that naive theories guided their inferences: negative motives were likely to be inferred from negative behaviors, whereas both negative and positive motives were inferred from positive behaviors. Implications for attribution theories and negativity bias in social perception are discussed.