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An ideal powder preparation technique results in minimum protein damage and yields particle sizes in the lower micrometre range and homogeneous particle size distribution enabling subcutaneous or intramuscular injection through hypodermic needles. As suspension vehicles traditional non-aqueous injectable liquids, such as plant oils, may be selected. But they show an inherent high viscosity, which can lead to unacceptable glide forces during injection. Furthermore, the vehicle should provide high product stability with respect to protein integrity and suspension resuspendability. This review will describe how proteins can be formulated as protein powder suspensions in non-aqueous vehicles for subcutaneous injection including potential vehicles, protein powder preparation techniques, protein and suspension physical stability, as well as the use in the field of high concentration protein formulations.The mammalian brain cortex is highly folded, with several developmental disorders affecting folding. On the extremes, lissencephaly, a lack of folds in humans, and polymicrogyria, an overly folded brain, can lead to severe mental retardation, short life expectancy, epileptic seizures, and tetraplegia. Not only a specific degree of folding, but also stereotyped patterns, are required for normal brain function. A quantitative model on how and why these folds appear during the development of the brain is the first step in understanding the cause of these conditions. In recent years, there have been various attempts to understand and model the mechanisms of brain folding. Previous works have shown that mechanical instabilities play a crucial role in the formation of brain folds, and that the geometry of the fetal brain is one of the main factors in dictating its folding characteristics. However, modeling higher-order folding, one of the main characteristics of the highly gyrencephalic brain, has not been fully tach are shallower and more variable than the structures obtained in earlier gyrification stage emerge, reproducing well-known characteristics of higher-order folding in the mammalian, and particularly the human, brain.We have previously shown that INS-fMRI is a rapid method for mapping mesoscale brain networks in the macaque monkey brain. Focal stimulation of single cortical sites led to the activation of connected cortical locations, resulting in a global connectivity map. Here, we have extended this method for mapping brainwide networks following stimulation of single subcortical sites. As a testbed, we focused on the basal nucleus of the amygdala in the macaque monkey. We describe methods to target basal nucleus locations with submillimeter precision, pulse train stimulation methods, and statistical tests for assessing non-random nature of activations. Using these methods, we report that stimulation of precisely targeted loci in the basal nucleus produced sparse and specific activations in the brain. Activations were observed in the insular and sensory association cortices as well as activations in the cingulate cortex, consistent with known anatomical connections. What is new here is that the activations were focal and, in some cases, exhibited shifting topography with millimeter shifts in stimulation site. The precision of the method enables networks mapped from different nearby sites in the basal nucleus to be distinguished. While further investigation is needed to improve the sensitivity of this method, our analyses do support the reproducibility and non-random nature of some of the activations. ABT-263 order We suggest that INS-fMRI is a promising method for mapping large-scale cortical and subcortical networks at high spatial resolution.

To improve the signal-to-noise ratio (SNR) and image sharpness for whole brain isotropic 0.5mm three-dimensional (3D) T

weighted (T

w) turbo spin echo (TSE) intracranial vessel wall imaging (IVWI) at 3T.

The variable flip angle (VFA) method enables useful optimization across scan efficiency, SNR and relaxation induced point spread function (PSF) for TSE imaging. A convolutional neural network (CNN) was developed to retrospectively enhance the acquired TSE image with PSF blurring. The previously developed VFA method to increase SNR at the expense of blur can be combined with the presented PSF correction to yield long echo train length (ETL) scan while the acquired image remains high SNR and sharp. The overall approach can enable an optimized solution for accelerated whole brain high-resolution 3D T

w TSE IVWI. Its performance was evaluated on healthy volunteers and patients.

The PSF blurred image acquired by a long ETL scan can be enhanced by CNN to restore similar sharpness as a short ETL scan, which outperforms the traditional linear PSF enhancement approach. For accelerated whole brain IVWI on volunteers, the optimized isotropic 0.5mm 3D T

w TSE sequence with CNN based PSF enhancement provides sufficient flow suppression and improved image quality. Preliminary results on patients further demonstrated its improved delineation for intracranial vessel wall and plaque morphology.

The CNN enhanced VFA TSE imaging enables an overall image quality improvement for high-resolution 3D T

w IVWI, and may provide a better tradeoff across scan efficiency, SNR and PSF for 3D TSE acquisitions.

The CNN enhanced VFA TSE imaging enables an overall image quality improvement for high-resolution 3D T1w IVWI, and may provide a better tradeoff across scan efficiency, SNR and PSF for 3D TSE acquisitions.In Japan, there are two species of scorpions, Madara scorpion (Isometrus maculatus) and Yaeyama scorpion (Liocheles australasiae), and both of them are living in Yaeyama island. It has been shown that Liocheles australasiae has venom including β-toxin acting on K+-channels (β-KTx) (Juichi et al., 2018) [1]. Interestingly, LaIT2, one of the toxins found in the venom of Liocheles australasiae, displays the virulence for insects but almost not for mammals. Until now, molecular mechanism of the functional specificity of LaIT2 is unknown. To clear this issue, we tried to establish the overexpression system of LaIT2 in Rosetta-gami B (DE3) pLysS, which have trxB/gor mutations to induce the disulfide bond formation. In this study, we have succeeded to overexpress the recombinant LaIT2 (rLaIT2) as a thioredoxin (Trx)-tagged protein, and established the purification protocol with Ni2+-NTA column chromatography, enterokinase digestion, and HPLC. We succeeded to obtain approximately 0.5 mg of rLaIT2 from the E. coli cells cultured in 1 L of M9 culture medium.

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