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In recent years, it has become an acknowledged fact that noise exposure can lead to cognitive impairments, and researchers have shown increasing interest in this area. However, the detrimental impact of noise exposure on Alzheimer's disease (AD) animal models might be considerably greater than on ordinary model mice, yet the mechanisms by which noise exposure affects the hippocampus in these models have been scarcely investigated. This study we used 4D Label-free proteomics to identify distinctive differentially expressed proteins in the hippocampus of AD model mice following noise exposure. Among these proteins, the presence of Cathepsin S(CTSS) cannot be disregarded. Utilizing experimental techniques such as Western blot, immunofluorescence, and rt-qPCR, we confirmed the expression of CTSS in the hippocampus of APP/PS1 mice after noise exposure. Additionally, we examined downstream molecules including P53,BCL-2, BAX, and CASPASE3 using KEGG pathway analysis. The results indicated an elevation in CTSS expression, a reduction in the anti-apoptotic gene BCL-2, and an increase in the expression of BAX and cleaved CASPASE3. Based on these findings, we hypothesize that noise exposure potentially heightens apoptosis within the hippocampus through upregulating CTSS expression, subsequently posing a threat to AD model animals. • This study represents the inaugural utilization of 4D label-free proteomics to explore the hippocampus changes in noise-exposed APP/PS1 mice. • Using 4D label-free proteomics, we found increased levels of CTSS protein in the hippocampus of noise-exposed APP/PS1 mice. • We have observed a link between CTSS protein upregulation and heightened hippocampal apoptosis in noise-exposed APP/PS1 mice. [ABSTRACT FROM AUTHOR]

Noise is an important environmental stressor in the industrialized world and has received increasing attention in recent years. Although epidemiological research has extensively demonstrated the relationship between noise and cognitive impairment, the specific molecular mechanisms and targets remain to be fully explored and understood. To address this issue, 5-month-old C57BL/6 mice were divided into two groups, with one group exposed to white noise at 98 dB. The effects of noise on cognition in mice were investigated through molecular biology and behavioral experiments. Subsequently, transcriptomic sequencing of the hippocampus in both groups of mice was performed and enrichment analysis of differentially expressed genes (DEGs) was conducted using KEGG and GO databases. Furthermore, LASSO analysis was used to further narrow down the relevant DEGs, followed by enrichment analysis of these genes using KEGG and GO databases. The DEGs were further validated by rt-qPCR. Following noise exposure, the hippocampus levels of inflammation-related factors increased, the phosphorylation of Tau protein increased, the postsynaptic density protein decreased, the number of Nissl bodies decreased, and cell shrinkage in the hippocampus increased. Moreover, the behavioral experiments manifest characteristics indicative of a decline in cognitive.A total of 472 DEGs were identified through transcriptomic analysis, and seven relevant genes were screened by the LASSO algorithm, which were further validated by PCR to confirm their consistency with the omics results. In conclusion, noise exposure affects cognitive function in mice through multiple pathways, and the omics results provide new evidence for the cognitive impairment induced by noise exposure. • Explored the impact of noise exposure on cognitive functions in C57BL/6 mice via behavioral and molecular biology analyses. • By transcriptomic analysis, we have scrutinized the alterations in the hippocampus of C57BL/6 mice following noise exposure. • Screened highly correlated molecules via LASSO analysis, offering new insights into noise exposure's impact on cognition. [ABSTRACT FROM AUTHOR]

The study aimed to observe the effects of noise exposure on the pericytes of the cochlear stria vascularis (SV) in mice and to investigate its molecular mechanism. Male C57BL/6J mice aged 6–8 weeks were used as the subjects. Auditory Brainstem Response (ABR) was used to assess hearing loss. Hematoxylin and Eosin (HE) staining was conducted to observe morphological alterations in the SV. Immunofluorescence combined with transmission electron microscopy (TEM) was used to scrutinize changes in pericytes following acoustic injury. Western blotting (WB) was used to assess the expression variations of the migration-related protein Osteopontin (OPN). Evans Blue assay was performed to evaluate the permeability of the blood labyrinth barrier (BLB). 4-Hydroxynonenal (4-HNE) staining, in conjunction with measurements of Superoxide Dismutase (SOD), Malondialdehyde (MDA), and Catalase (CAT) content, was used to ascertain whether oxidative stress injury occurred in the SV. WB, combined with immunofluorescence, was used to examine alterations in the expression of proliferator-activated receptor-gamma coactivator 1α (PGC-1α) in the SV and pericytes. Noise exposure resulted in permanent hearing loss in C57BL/6J mice, accompanied by SV swelling, migration of pericytes from their vascular attachments, BLB leakage, elevated oxidative stress levels in the SV, and reduced expression of PGC-1α on both the SV and migrating pericytes. Noise exposure may potentially increase oxidative stress levels in the SV, downregulate the expression levels of PGC-1α, promote pericytes migration, and subsequently lead to an elevation in BLB permeability. • This study has unveiled potential association between PGC-1α and pericyte migration. • This study has elucidated the changes in OPN expression and suggested a potential role of OPN in pericyte migration. • This study has illuminated the impact of oxidative stress-induced damage on the stria vascularis after noise exposure. [ABSTRACT FROM AUTHOR]

[Display omitted] • HLM is a method that classifies helical filaments using 2D classification labels. • HLM is the first method that applies language models to cryo-EM image processing. • HLM is validated with many simulation tests and experimental cryo-EM datasets in EMPIAR. • The Transformer-based pipeline is more robust than the traditional word2vec pipeline. • HLM tests led to the discovery of a novel amyloid variant. Many macromolecules in biological systems exist in the form of helical polymers. However, the inherent polymorphism and heterogeneity of samples complicate the reconstruction of helical polymers from cryo-EM images. Currently, available 2D classification methods are effective at separating particles of interest from contaminants, but they do not effectively differentiate between polymorphs, resulting in heterogeneity in the 2D classes. As such, it is crucial to develop a method that can computationally divide a dataset of polymorphic helical structures into homogenous subsets. In this work, we utilized deep-learning language models to embed the filaments as vectors in hyperspace and group them into clusters. Tests with both simulated and experimental datasets have demonstrated that our method – HLM (H elical classification with L anguage M odel) can effectively distinguish different types of filaments, in the presence of many contaminants and low signal-to-noise ratios. We also demonstrate that HLM can isolate homogeneous subsets of particles from a publicly available dataset, resulting in the discovery of a previously unreported filament variant with an extra density around the tau filaments. [ABSTRACT FROM AUTHOR]

Branched-chain fatty acids (BCFAs) are key precursors of branched-chain fuels, which have cold-flow properties superior to straight chain fuels. BCFA production in Gram-negative bacterial hosts is inherently challenging because it competes directly with essential and efficient straight-chain fatty acid (SCFA) biosynthesis. Previously, Escherichia coli strains engineered for BCFA production also co-produced a large percentage of SCFA, complicating efficient isolation of BCFA. Here, we identified a key bottleneck in BCFA production: incomplete lipoylation of 2-oxoacid dehydrogenases. We engineered two protein lipoylation pathways that not only restored 2-oxoacid dehydrogenase lipoylation, but also increased BCFA production dramatically. E. coli expressing an optimized lipoylation pathway produced 276 mg/L BCFA, comprising 85% of the total free fatty acids (FFAs). Furthermore, we fine-tuned BCFA branch positions, yielding strains specifically producing ante-iso or odd-chain iso BCFA as 77% of total FFA, separately. When coupled with an engineered branched-chain amino acid pathway to enrich the branched-chain α-ketoacid pool, BCFA can be produced from glucose at 181 mg/L and 72% of total FFA. While E. coli can metabolize BCFAs, we demonstrated that they are not incorporated into the cell membrane, allowing our system to produce a high percentage of BCFA without affecting membrane fluidity. Overall, this work establishes a platform for high percentage BCFA production, providing the basis for efficient and specific production of a variety of branched-chain hydrocarbons in engineered bacterial hosts. [ABSTRACT FROM AUTHOR]

A stochastic averaging method is proposed for nonlinear vibration energy harvesters subject to Gaussian white noise excitation. The generalized harmonic transformation scheme is applied to decouple the electromechanical equations, and then obtained an equivalent nonlinear system which is uncoupled to an electric circuit. The frequency function is given through the equivalent potential energy which is independent of the total energy. The stochastic averaging method is developed by using the generalized harmonic functions. The averaged Itô equations are derived via the proposed procedure, and the Fokker-Planck-Kolmogorov (FPK) equations of the decoupled system are established. The exact stationary solution of the averaged FPK equation is used to determine the probability densities of the amplitude and the power of the stationary response. The procedure is applied to three different type Duffing vibration energy harvesters under Gaussian white excitations. The effects of the system parameters on the mean-square voltage and the output power are examined. It is demonstrated that quadratic nonlinearity only and quadratic combined with properly cubic nonlinearities can increase the mean-square voltage and the output power, respectively. The approximate analytical outcomes are qualitatively and quantitatively supported by the Monte Carlo simulations. [ABSTRACT FROM AUTHOR]

Abstract: Snap-through mechanism is employed to harvest electricity from random vibration through piezoelectricity. The random excitation is assumed to be Gaussian white noise. The snap-through piezoelectric energy harvester possesses the bistability. For small-amplitude vibration in a potential well, the Ito stochastic differential equation of the electromechanical coupling system is derived from the Taylor approximation at a stable equilibrium point. The method of the moment differential equations is applied to determine the statistical moments of the displacement response and the output voltage. The effects of the system parameters on the output voltage and the output power are examined. The approximate analytical outcomes are qualitatively and quantitatively supported by the numerical simulations. For large-amplitude interwell motion, the effects of the parameters on the output voltage and the output power are numerically investigated. Nonlinearity produced by the snap-through improves energy harvesting so that the snap-through piezoelectric energy harvester can outperform the linear energy harvester in the similar size under Gaussian white noise excitations. [Copyright &y& Elsevier]

Abstract: Accurate determination of micrograph focuses is essential for averaging multiple images to reach high-resolution 3-D reconstructions in electron cryo-microscopy (cryo-EM). Current methods use iterative fitting of focus-dependent simulated power spectra to the power spectra of experimental images, with the fitting performed independently for different images. Here we have developed a novel graph theory based method in which the rotational average focus and individual angular sector focuses of all images are determined simultaneously in closed form using the least square solution of overdetermined linear equations. The new method was shown to be fast, accurate, and robust in tests with large datasets of experimental low dose cryo-EM images. Its integration with three classic power spectra fitting methods also allows cross validation of the results by these vastly different methods. The new integrated focus determination method will improve reliability of automated focus determination for large-scale data processing that is increasingly common in the cryo-EM field. [Copyright &y& Elsevier]

Abstract: Toxoplasma gondii (T. gondii) is a unicellular protozoan that infects up to one-third of the world''s human population. Numerous studies revealed that a latent infection of T. gondii can cause life-threatening encephalitis in immunocompromised people and also has significant effects on the behavior of healthy people and animals. However, the overall transmission of T. gondii has not been well understood although many factors affecting this process have been found out by different biologists separately. Here we synthesize what is currently known about the natural history of T. gondii by developing a prototype agent-based model to mimic the transmission process of T. gondii in a farm system. The present model takes into account the complete life cycle of T. gondii, which includes the transitions of the parasite from cats to environment through feces, from contaminated environment to mice through oocysts, from mice to cats through tissue cysts, from environment to cats through oocysts as well as the vertical transmission among mice. Although the current model does not explicitly include humans and other end-receivers, the effect of the transition to end-receivers is estimated by a developed infection risk index. The current model can also be extended to include human activities and thus be used to investigate the influences of human management on disease control. Simulation results reveal that most cats are infected through preying on infected mice while mice are infected through vertical transmission more often than through infection with oocysts, which clearly suggests the important role of mice during the transmission of T. gondii. Furthermore, our simulation results show that decreasing the number of mice on a farm can lead to the eradication of the disease and thus can lower the infection risk of other intermediate hosts on the farm. In addition, with the assumption that the relation between virulence and transmission satisfies a normal function, we show that intermediate virulent lineages (type II) can sustain the disease most efficiently, which can qualitatively agree with the fact that the evolution of the parasite favors intermediate virulence. The effects of other related factors on transmission, including the latent period and imprudent behavior of mice, and prevention strategies are also studied based on the present model. [Copyright &y& Elsevier]

Abstract: In this article, a conspicuously simple and highly sensitive amperometric immunosensor based on the sequential electrodeposition of Prussian blue (PB) and gold nanoparticles (GNPs) on multiwalled carbon nanotube (MWCNT)-modified glassy carbon electrode (GCE) surface is proposed for the detection of α-fetoprotein (AFP). By comparison with PB, the MWCNT/PB composite film had been proven to show much better electrochemical stability and a larger response current. The electrodeposited GNP film can be used not only to immobilize biomolecules but also to avoid the leakage of PB and to prevent shedding of MWCNT/PB composite film from the electrode surface. The performance and factors influencing the performance of the immunosensor were investigated. Under optimal experimental conditions, the proposed immunosensor for AFP was observed with an ultralow limit of detection (LOD) equal to 3pg/ml (at 3δ), and the linear working range spanned the concentrations of AFP from 0.01 to 300ng/ml. Moreover, the immunosensor, as well as a commercially available kit, was examined for use in the determination of AFP in real human serum specimens. More significant, the assay mentioned here is simpler than the traditional enzyme-linked immunosorbent assay (ELISA), and an excellent correlation of levels of AFP measured was obtained, indicating that the developed immunoassay could be a promising alternative approach for detection of AFP and other tumor markers in the clinical diagnosis. [ABSTRACT FROM AUTHOR]

Abstract: Reduced glutathione (GSH) is the most powerful intracellular antioxidant and also involved in viral infections. The pathogenesis of dengue virus (DV) infection has not been completely clarified. This study investigated the relationship between DV serotype 2 (DV2) infections and host intracellular GSH content. Results showed infection with DV2 resulted in a decrease in intracellular GSH, which caused NF-κB activation and increased DV2 production. Supplemental GSH significantly inhibited activation of NF-κB, resulting in a decreased production of DV2 in HepG2 cells. Furthermore, high activity of NF-κB and increased production of DV2 was observed in HepG2 cells treated with buthionine sulfoximine (BSO), an inhibitor of GSH synthesis. In conclusion, DV2 infection could reduce host intracellular GSH concentration and benefited from this process. Supplemental GSH could inhibit viral production, indicating GSH might be valuable in the prevention and treatment of DV2 infection. [Copyright &y& Elsevier]

Abstract: In this study, superparamagnetic monodisperse magnetite colloids, around 5nm in size, were prepared by dissolving iron chlorides, sodium hydroxide (NaOH) and sodium oleate (NaOL), in toluene/ethanol/water mixtures and refluxing for 4h. The concentrations of NaOH and NaOL were varied to systematically investigate the effect on the surface properties, size, dispersion, and magnetic properties of magnetite nanoparticles (MNPs). The samples were characterized via XRD, FTIR, TGA, TEM, SAED, DLS, and VSM. The results indicated that the surface coatings of MNPs could be manipulated from oleate to hydroxyl groups via increasing the molar ratio of NaOH/Fe(II) more than 8. The amount of NaOH had no obvious influence on the size and the saturation magnetization of MNPs. Therefore NaOH was not a necessary reactant for forming magnetite crystals. On the contrary, NaOL was shown to be the most important component for synthesizing stable magnetite colloids. The NaOL acted as both a key reactant to buffer the pH environment and a surfactant to keep the MNPs stable in nonpolar solvent media. [Copyright &y& Elsevier]

Abstract: Dicamba (3,6-dichloro-2-methoxybenzoic acid) is a widely used herbicide that is efficiently degraded by soil microbes. These microbes use a novel Rieske nonheme oxygenase, dicamba monooxygenase (DMO), to catalyze the oxidative demethylation of dicamba to 3,6-dichlorosalicylic acid (DCSA) and formaldehyde. We have determined the crystal structures of DMO in the free state, bound to its substrate dicamba, and bound to the product DCSA at 2.10–1.75 Å resolution. The structures show that the DMO active site uses a combination of extensive hydrogen bonding and steric interactions to correctly orient chlorinated, ortho-substituted benzoic-acid-like substrates for catalysis. Unlike other Rieske aromatic oxygenases, DMO oxygenates the exocyclic methyl group, rather than the aromatic ring, of its substrate. This first crystal structure of a Rieske demethylase shows that the Rieske oxygenase structural scaffold can be co-opted to perform varied types of reactions on xenobiotic substrates. [Copyright &y& Elsevier]

Abstract: Prostate transglutaminase (TGase-4 or TGaseP) is an enzyme that is uniquely expressed in prostate tissues. The function of the TGase, implicated in the cell-matrix, is yet to be fully established. In the present study, we investigated the role of TGase-4 in tumor-endothelial cell interactions, by creating a panel of prostate cancer cell lines that have different expression profiles of human TGase-4. Here, we report that prostate cancer cells PC-3, when over-expressing TGase-4 (PC-3TGase4exp) increased their ability to adhere to quiescent and activated (by hepatocyte growth factor) endothelial cells. In contrast, the prostate cancer cell CAHPV-10, which expressed high levels of TGase-4, reduced the adhesiveness to the endothelial cells after TGase-4 expression was knocked down. By using frequency based electric cell impedance sensing, we found that TGase-4 mediated adhesion resulted in a change in impedance at low frequency (400 Hz), indicating a paracellular pathway disruption. The study further showed that expression of TGase-4 rendered the cells to exert regulation of endothelial interaction by bypassing the ROCK pathway. It is therefore concluded, that TGase-4 plays a pivotal role in the interaction between endothelial cells and prostate cancer cells, an action which is independent of the ROCK pathway. [Copyright &y& Elsevier]

Abstract: Accurately determining a cryoEM particle’s alignment parameters is crucial to high resolution single particle 3-D reconstruction. We developed Multi-Path Simulated Annealing, a Monte-Carlo type of optimization algorithm, for globally aligning the center and orientation of a particle simultaneously. A consistency criterion was developed to ensure the alignment parameters are correct and to remove some bad particles from a large pool of images of icosahedral particles. Without using any a priori model, this procedure is able to reconstruct a structure from a random initial model. Combining the procedure above with a new empirical double threshold particle selection method, we are able to pick tens of best quality particles to reconstruct a subnanometer resolution map from scratch. Using the best 62 particles of rice dwarf virus, the reconstruction reached 9.6Å resolution at which four helices of the P3A subunit of RDV are resolved. Furthermore, with the 284 best particles, the reconstruction is improved to 7.9Å resolution, and 21 of 22 helices and six of seven β sheets are resolved. [Copyright &y& Elsevier]

Abstract: The HCV polymerase is an attractive target for the development of new and specific anti-HCV drugs. Herein, the characterization of the inhibitory effect of 2′-C-Methyl-Cytidine shows that it is a potent inhibitor of both genotype 1b and 1a HCV replicon replication, both of laboratory-optimized as well as of NS5B clinical isolates–chimera replicons. The corresponding 5′-triphosphate derivative is a potent inhibitor of native HCV replicase isolated from replicon cells and of the recombinant genotype 1b and 1a HCV polymerase-mediated RNA synthesis. Resistance to 2′-C-Methyl-Cytidine was mapped to amino acid substitution S282T in the NS5B coding region. Cross-resistance was observed to 2′-C-Methyl-Adenosine but not to interferon α-2a, to non-nucleoside HCV polymerase inhibitors or to R1479, a new and potent nucleoside inhibitor of NS5B polymerase. In vitro studies mapped resistance to R1479 to amino acid substitutions S96T and S96T/N142T of the NS5B polymerase. These mutations did not confer resistance to 2-C-Methyl-Cytidine, thus confirming the lack of cross-resistance between these two HCV inhibitors. These data will allow the optimization of new polymerase inhibitors and their use in combination therapy. [Copyright &y& Elsevier]

Abstract: VEGF receptor-2 plays a critical role in endothelial cell proliferation during angiogenesis. However, regulation of receptor activity remains incompletely explained. Here, we demonstrate that VEGF stimulates microvascular endothelial cell proliferation in a dose-dependent manner with VEGF-induced proliferation being greatest at 5 and 100 ng/ml and significantly reduced at intermediate concentrations (>50% at 20 ng/ml). Neutralization studies confirmed that signaling occurs via VEGFR-2. In a similar fashion, ERK/MAPK is strongly activated in response to VEGF stimulation as demonstrated by its phosphorylation, but with a decrease in phosphoryation at 20 ng/ml VEGF. Immunoblotting analysis revealed that VEGF did not cause a dose-dependent change in expression of VEGFR-2 but instead resulted in reduced phosphorylation of VEGFR-2 when cells were exposed to 10 and 20 ng/ml of VEGF. VEGFR-2 dephosphorylation was associated with an increase in the protein tyrosine phosphatase, SH-PTP1, and endothelial nitric oxide synthase (eNOS). Immunoprecipitation and selective immunoblotting confirmed the association between VEGFR-2 dephosphorylation and the upregulation of SH-PTP1 and eNOS. Transfection of endothelial cells with antisense oligonucleotide against VEGFR-2 completely abolished VEGF-induced proliferation, whereas anti SH-PTP1 dramatically increased VEGF-induced proliferation by 1 and 5-fold at 10 and 200 ng/ml VEGF, respectively. Suppression of eNOS expression only abolished endothelial cell proliferation at VEGF concentrations above 20 ng/ml. Taken together, these results indicate that activation of VEGFR-2 by VEGF enhances SH-PTP1 activity and eNOS expression, which in turn lead to two diverse events: one is that SH-PTP1 dephosphorylates VEGFR-2 and ERK/MAPK, which weaken VEGF mitogenic activity, and the other is that eNOS increases nitric oxide production which in turn lowers SH-PTP1 activity via S-nitrosylation. [Copyright &y& Elsevier]

Abstract: The presence of noise and absence of contrast in electron micrographs lead to a reduced resolution of the final 3D reconstruction, due to the inherent limitations of single-particle image alignment. The fast rotational matching (FRM) algorithm was introduced recently for an accurate alignment of 2D images under such challenging conditions. Here, we implemented this algorithm for the first time in a standard 3D reconstruction package used in electron microscopy. This allowed us to carry out exhaustive tests of the robustness and reliability in iterative orientation determination, classification, and 3D reconstruction on simulated and experimental image data. A classification test on GroEL chaperonin images demonstrates that FRM assigns up to 13% more images to their correct reference orientation, compared to the classical self-correlation function method. Moreover, at sub-nanometer resolution, GroEL and rice dwarf virus reconstructions exhibit a remarkable resolution gain of 10–20% that is attributed to the novel image alignment kernel. [Copyright &y& Elsevier]

Abstract: Background: Com-1 is a molecule that has recently discovered to have putative action on the metastatic nature of cancer cells. The current study investigated the impact of Com-1 on oestrogen regulated cell growth of breast cancer cells and explored the potential link between Com-1 and ER-β. Method: Full length Com-1 cDNA was isolated from normal mammary tissues. Ribozyme transgenes that specifically targeted human Com-1 were constructed using the pEF6/V5-His vector. Expression of Com-1 was assessed at both mRNA and protein levels. Interaction of Com-1 with other candidate molecules was studied using immunoprecipitation and Western blotting. Results: Elimination of Com-1 by way of ribozyme transgenes results in breast cancer cells with increased rate of growth and increased invasive potential. In contrast, over-expression of Com-1 in the cancer cells had an opposite effect. In ER-α-negative/ER-β positive MDA MB-231 cells, elimination of Com-1 caused more vigorous growth in response to 17-β-estradiol. However, the effect of Com-1 modification on MCF-7, which is positive for both ER-α and ER-β, was less clear. Protein interaction analysis has indicated that the Com-1 and ER-β were mutually co-precipitated with each other in breast cancer cells. Immunocytochemical staining revealed that Com-1 was primarily present in the nucleus, with some degree of cytoplasmic staining, and that the distribution of Com-1 was identical to that of ER-β. 17-β-Estradiol stimulation resulted in reduction of nucleic staining of Com-1. This reduction of nucleic Com-1 can be reverted when ubiquitin inhibitor, ubiquitin aldehyde or the lactacystin proteosome inhibitor was present, suggesting a pivotal role of the ubiquitin–proteosome pathway in the Com-1/ER-β complex. Conclusion: Com-1 plays a tumour suppressor role in breast cancer cells and is involved in oestrogen-regulated cell growth. This action is potentially exerted by interacting with ER-β, in human breast cancer cells. The fate of Com-1 can be dually regulated by oestrogen and ubiquitin pathway. [Copyright &y& Elsevier]

Sub-nanometer resolution structure determination is becoming a common practice in electron cryomicroscopy of macromolecular assemblies. The data for these studies have until now been collected on photographic film. Using cytoplasmic polyhedrosis virus (CPV), a previously determined structure, as a test specimen, we show the feasibility of obtaining a 9 Å structure from images acquired from a 4 k × 4 k Gatan CCD on a 200 kV electron cryomicroscope. The match of the α-helices in the protein components of the CPV with the previous structure of the same virus validates the suitability of this type of camera as the recording media targeted for single particle reconstructions at sub-nanometer resolution. [Copyright &y& Elsevier]

Treatment of 3T3-L1 preadipocytes with differentiation inducers triggers a cascade in which C/EBPβ is rapidly expressed, followed by C/EBPα and PPARγ. C/EBPα and PPARγ then activate the expression of adipocyte genes that produce the differentiated phenotype. Circumstantial evidence indicates that C/EBPβ activates transcription of the C/EBPα and PPARγ genes, both of which possess C/EBP regulatory elements in their proximal promoters. Although C/EBPβ is expressed immediately upon induction of differentiation, acquisition of DNA binding activity is delayed for ∼14 h. Chromatin immunoprecipitation (ChIP) analysis conducted 24 h after induction revealed that C/EBPβ binds to C/EBP regulatory elements in the proximal promoters of the C/EBPα and PPARγ genes. After an additional delay ChIP analysis showed that C/EBPα binds to its own promoter and to the promoters of the PPARγ and 422/aP2 genes. These findings support the view that once expressed, C/EBPα is responsible for maintaining the expression of PPARγ and C/EBPα, as well as adipocyte proteins (e.g., 422/aP2) in the terminally differentiated state. Together these findings provide compelling evidence that C/EBPβ, C/EBPα, and PPARγ participate in a cascade during adipogenesis. [Copyright &y& Elsevier]

Treatment of 3T3-L1 preadipocytes with differentiation inducers triggers a cascade in which C/EBPβ is rapidly expressed, followed by C/EBPα and PPARγ. C/EBPα and PPARγ then activate the expression of adipocyte genes that produce the differentiated phenotype. Circumstantial evidence indicates that C/EBPβ activates transcription of the C/EBPα and PPARγ genes, both of which possess C/EBP regulatory elements in their proximal promoters. Although C/EBPβ is expressed immediately upon induction of differentiation, acquisition of DNA binding activity is delayed for ∼14 h. Chromatin immunoprecipitation (ChIP) analysis conducted 24 h after induction revealed that C/EBPβ binds to C/EBP regulatory elements in the proximal promoters of the C/EBPα and PPARγ genes. ChIP analysis showed that after an additional delay C/EBPα binds to its own promoter and to the promoters of the PPARγ and 422/aP2 genes. These findings support the view that once expressed, C/EBPα is responsible for maintaining the expression of PPARγ, and C/EBPα, as well as adipocyte proteins (e.g., 422/aP2) in the terminally differentiated state. Together these findings provide compelling evidence that C/EBPβ, C/EBPα, and PPARγ participate in a cascade during adipogenesis. [Copyright &y& Elsevier]

Due to the sensitivity of biological sample to the radiation damage, the low dose imaging conditions used for electron microscopy result in extremely noisy images. The processes of digitization, image alignment, and 3D reconstruction also introduce additional sources of noise in the final 3D structure. In this paper, we investigate the effectiveness of a bilateral denoising filter in various biological electron microscopy applications. In contrast to the conventional low pass filters, which inevitably smooth out both noise and structural features simultaneously, we found that bilateral filter holds a distinct advantage in being capable of effectively suppressing noise without blurring the high resolution details. In as much, we have applied this technique to individual micrographs, entire 3D reconstructions, segmented proteins, and tomographic reconstructions. [Copyright &y& Elsevier]

The dispositions of 39 α helices of greater than 2.5 turns and four β sheets in the major capsid protein (VP5, 149 kDa) of herpes simplex virus type 1 were identified by computational and visualization analysis from the 8.5 A˚ electron cryomicroscopy structure of the whole capsid. The assignment of helices in the VP5 upper domain was validated by comparison with the recently determined crystal structure of this region. Analysis of the spatial arrangement of helices in the middle domain of VP5 revealed that the organization of a tightly associated bundle of ten helices closely resembled that of a domain fold found in the annexin family of proteins. Structure-based sequence searches suggested that sequences in both the N and C-terminal portions of the VP5 sequence contribute to this domain. The long helices seen in the floor domain of VP5 form an interconnected network within and across capsomeres. The combined structural and sequence-based informatics has led to an architectural model of VP5. This model placed in the context of the capsid provides insights into the strategies used to achieve viral capsid stability. [Copyright &y& Elsevier]

Abstract: A simple, inexpensive microdistillation device is described for capturing methanol or formaldehyde as end products of biochemical reactions or in environmental samples. We demonstrate that the microdistillation protocol, coupled with the use of alcohol oxidase and the formaldehyde-sensitive reagent Purpald (4-amino-3-hydrazino-5-mercapto-1,2,4-triazole), serves as a quick and inexpensive alternative to chromatographic and mass spectrometer analyses for determining if formaldehyde or methanol is a product of reactions that contain substances that interfere with the Purpald reaction. These techniques were used to affirm formaldehyde as the end product of the dicamba monooxygenase-catalyzed O-demethylation of the herbicide dicamba (2-methoxy-3,6-dichlorobenzoic acid). [Copyright &y& Elsevier]

With the continuous growth of the global population and the increasing demand for natural resources, ecological resource scarcity has become a global issue that cannot be ignored. However, little is known about individuals' environment-related responses to ecological resource scarcity. Will people save resources for the future or consume them for immediate interests when resources become scarce? Drawing on the life history theory and construal level theory, we propose that individuals' environment-related responses to ecological resource scarcity depend on levels of future orientation. Analyses of multilevel data from 30 countries found that only in highly future-oriented countries could objective ecological resource scarcity positively predict individuals' daily pro-environmental behaviors (Study 1). Two subsequent studies at the individual level again established the moderating effect of future orientation on the link between perceived ecological resource scarcity and pro-environmental behaviors (Studies 2 & 3), and the causal effect was also clarified (Study 3). Specifically, only for future-oriented people, perceived ecological resource scarcity or reminders of it could positively promote pro-environmental behaviors. We further verified the moderation hypothesis with water-saving posters to explore potential practical implications, i.e., whether the poster with water shortage information would have an effective promotional effect only when information about "concern for the future" is presented (Study 4). We concluded with a discussion of the theoretical and practical implications of the research findings. • Exposure to ecological resource scarcity can't always promote individuals' pro-environmental behaviors. • The positive objective ecological resource scarcity-behavior link emerged only in countries with future-oriented culture. • The positive perceived ecological resource scarcity-behavior link emerged only for individuals with future-oriented traits. • Poster with water shortage information had a better publicity effect when future-oriented information was provided. [ABSTRACT FROM AUTHOR]

Abstract: Cardiovascular complications account for significant morbidity and mortality in the diabetic population. Diabetic cardiomyopathy (DCM), a prominent cardiovascular complication, has been recognized as a microvascular disease that may lead to heart failure. During the past few decades, research progress has been made in investigating the pathophysiology of the disease; however, the exact molecular mechanism has not been elucidated, making therapeutic a difficult task. In this review article, we have discussed a number of diabetes-induced metabolites such as glucose, advanced glycation end products, protein kinase C, free fatty acid and oxidative stress and other related factors that are implicated in the pathophysiology of the DCM. An understanding of the biochemical and molecular changes especially early in the DCM may lead to new and effective therapies toward prevention and amelioration of DCM, which is important for the millions of individuals who already have or are likely to develop the disease before a cure becomes available. [Copyright &y& Elsevier]

Service composition is a research hotspot with practical value. With the development of Web service, many Web services with the same functional attributes emerge. However, service composition optimization is still a big challenge since the complex and unstable composition environment. To solve this problem, we propose an adaptive service composition based on deep reinforcement learning, where recurrent neural network (RNN) is utilized for predicting the objective function, improving its expression and generalization ability, and effectively solving the shortcomings of traditional reinforcement learning in the face of large-scale or continuous state space problems. We leverage heuristic behavior selection strategy to divide the state set into hidden state and fully visible state. Effective simulation of hidden state space and fully visible state of the evaluation function can further improve the accuracy and efficiency of the combined results. We conduct comprehensive experiment and experimental results have shown the effectiveness of our method. [ABSTRACT FROM AUTHOR]

[Display omitted] • Activated concave APF nanospheres with high surface area and abundant functional groups (e.g., amines, imines) were successfully synthesized. • The efficient uptake of Cr(VI) and Cd(II) ions from aqueous medium were 1054 and 342 mg g−1, respectively. • Approximately 71.5% of Cr(VI) was simultaneously reduced to Cr(III). • The adsorption mechanism and reduction mechanism were explored. The issue of heavy metal contamination in water is a global concern, and the development of highly efficient adsorbent materials is crucial for the removal and detoxification of heavy metals. Polymer-based materials have emerged as a promising class of adsorbents due to their ability to capture heavy metal pollutants and reduce them to less toxic forms. The limited surface area of conventional polymer adsorbents makes them less effective for high-capacity adsorption. Herein, we present a low-temperature steam activation approach to address this challenge. This activation approach leads to a remarkable increase of over 20 times in the surface area of concave aminophenol-formaldehyde (APF) polymer nanospheres (from 45 to 961 m2/g) while preserving their reductive functional groups. The activated concave APF nanospheres were evaluated for their adsorption capabilities towards two typical heavy metal ions (i.e., Cr(VI) and Cd(II)) in aqueous solutions. The maximum adsorption capacities achieved were 1054 mg g−1 for Cr(VI) and 342 mg g−1 for Cd(II), which are among the highest performances reported in the literature and are much higher than the capacities of the non-activated APF nanospheres. Additionally, approximately 71.5 % of Cr(VI) was simultaneously reduced to Cr(III) through the benzenoid amine pathway during adsorption, highlighting the crucial role of the steam activation strategy in enhancing the capability of polymer adsorbents. [ABSTRACT FROM AUTHOR]

[Display omitted] Tumor microenvironment (TME) stimuli-responsive nanoassemblies are emerging as promising drug delivery systems (DDSs), which acquire controlled release by structural transformation under exogenous stimulation. However, the design of smart stimuli-responsive nanoplatforms integrated with nanomaterials to achieve complete tumor ablation remains challenging. Therefore, it is of utmost importance to develop TME-based stimuli-responsive DDSs to enhance drug-targeted delivery and release at tumor sites. Herein, we proposed an appealing strategy to construct fluorescence-mediated TME stimulus-responsive nanoplatforms for synergistic cancer therapy by assembling photosensitizers (PSs) carbon dots (CDs), chemotherapeutic agent ursolic acid (UA), and copper ions (Cu2+). First, UA nanoparticles (UA NPs) were prepared by self-assembly of UA, then UA NPs were assembled with CDs via hydrogen bonding force to obtain UC NPs. After combining with Cu2+, the resulting particles (named UCCu2+ NPs) exhibited quenched fluorescence and photosensitization due to the aggregation of UC NPs. Upon entering the tumor tissue, the photodynamic therapy (PDT) and the fluorescence function of UCCu2+ were recovered in response to TME stimulation. The introduction of Cu2+ triggered the charge reversal of UCCu2+ NPs, thereby promoting lysosomal escape. Furthermore, Cu2+ resulted in additional chemodynamic therapy (CDT) capacity by reacting with hydrogen peroxide (H 2 O 2) as well as by consuming glutathione (GSH) in cancer cells through a redox reaction, hence magnifying intracellular oxidative stress and enhancing the therapeutic efficacy due to reactive oxygen species (ROS) therapy. In summary, UCCu2+ NPs provided an unprecedented novel approach for improving the therapeutic efficacy through the three-pronged (chemotherapy, phototherapy, and heat-reinforced CDT) attacks to achieve synergistic therapy. [ABSTRACT FROM AUTHOR]

The recent outbreak of Corona Virus Disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been a severe threat to the global public health and economy, however, effective drugs to treat COVID-19 are still lacking. Here, we employ a deep learning-based drug repositioning strategy to systematically screen potential anti-SARS-CoV-2 drug candidates that target the cell entry mechanism of SARS-CoV-2 virus from 2635 FDA-approved drugs and 1062 active ingredients from Traditional Chinese Medicine herbs. In silico molecular docking analysis validates the interactions between the top compounds and host receptors or viral spike proteins. Using a SARS-CoV-2 pseudovirus system, we further identify several drug candidates including Fostamatinib, Linagliptin, Lysergol and Sophoridine that can effectively block the cell entry of SARS-CoV-2 variants into human lung cells even at a nanomolar scale. These efforts not only illuminate the feasibility of applying deep learning-based drug repositioning for antiviral agents by targeting a specified mechanism, but also provide a valuable resource of promising drug candidates or lead compounds to treat COVID-19. • A host-directed drug repurposing targeting the cell entry mechanism of SARS-CoV-2. • Three independent deep learning methods for drug repurposing. • Thousands of approved drugs and natural compounds for repurposing. • Validation of top drug candidates by molecular docking and virus entry assay. [ABSTRACT FROM AUTHOR]

Abstract Porous materials are effective for the isolation of sound with medium to high frequencies, while periodic structures are promising for low to medium frequencies. In the present work, we study the sound insulation of a periodically rib-stiffened double-panel with porous lining to reveal the effect of combining the two characters above. The theoretical development of the periodic composite structure, which is based on the space harmonic series and Biot theory, is included. The system equations are subsequently solved numerically by employing a precondition method with a truncation procedure. This theoretical and numerical framework is validated with results from both theoretical and finite element methods. The parameter study indicates that the presence of ribs can lower the overall sound insulation, although a direct transfer path is absent. Despite the unexpected model results, the method proposed here, which combines poroelastic modeling and periodic structures semi-analytically, can be promising in broadband sound modulation. Highlights • The effect of combining periodic structure and porous media is studied. • A method combining Biot theory with space harmonic series is developed and used. • The periodically rib-stiffened cases are analyzed subsequently. • This semi-analytical technique can be promising in broadband sound modulation. [ABSTRACT FROM AUTHOR]

An important way to promote the environmental industry's goal of carbon reduction is to promote the recycling of resources. Membrane separation technology has unique advantages in resource recovery and advanced treatment of industrial wastewater. However, the great promise of traditional organic membrane is hampered by challenges associated with organic solvent tolerance, lack of oxidation resistance, and serious membrane fouling control. Moreover, the high concentrations of organic matter and inorganic salts in the membrane filtration concentrate also hinder the wider application of the membrane separation technology. The emerging cost-effective graphene oxide (GO)-based membrane with excellent resistance to organic solvents and oxidants, more hydrophilicity, lower membrane fouling, better separation performance has been expected to contribute more in industrial wastewater treatment. Herein, we provide comprehensive insights into the preparation and characteristic of GO membranes, as well as current research status and problems related to its future application in industrial wastewater treatment. Finally, concluding remarks and future perspectives have been deduced and recommended for the GO membrane separation technology application for industrial wastewater treatment, which leads to realizing sustainable wastewater recycling and a nearly "zero discharge" water treatment process. [Display omitted] • Detailed description of the demand for industrial wastewater advanced treatment. • The characteristics and performance of GO membrane in advanced treatment of industrial wastewater were described. • The innovative application of GO membrane in advanced treatment of industrial wastewater was summarized. • How to deal with membrane fouling of GO membrane in industrial wastewater treatment was discussed in detail. • The application potential of GO membrane in industrial wastewater treatment was prospected in detail. [ABSTRACT FROM AUTHOR]

Minimization of the astigmatism of the objective lens is a critical daily instrument alignment task essential for high resolution TEM imaging. Fast and sensitive detection of astigmatism is needed to provide real-time feedback and adjust the stigmators to efficiently reduce astigmatism. Currently the method used by many microscopists is to visually examine the roundness of a diffractogram (Thon rings) and iteratively adjust the stigmators to make the Thon rings circular. This subjective method is limited by poor sensitivity and potentially biased by the astigmatism of human eyes. In this study, an s 2 power spectra based method, s 2 stigmator , was developed to allow fast and sensitive detection of the astigmatism in TEM live images. The “radar”-style display provides real-time feedback to guide the adjustment of the objective lens stigmators. Such unique capability allowed us to discover the mapping of the two stigmators to the astigmatism amplitude and angle, which led us to develop a single-pass tuning strategy capable of significantly quicker minimization of the objective lens astigmatism. [ABSTRACT FROM AUTHOR]

An 8-weeks feeding trial was conducted to estimate the effects of different viscous cellulose on the intestinal flora and health in juvenile largemouth bass (Micropterus salmoides). Four isoproteic and isolipidic experimental diets were formulated (crude protein 42.50%, crude lipid 13.70%) to contain 8% cellulose (control group; 5.14 mPa s), 8% low viscous carboxymethyl cellulose (CMC) with 800 mPa s (Lvs-CMC group; 182.15 mPa s), 8% middle viscous CMC with 2000 mPa s (Mvs-CMC group; 320.48 mPa s) and 8% high viscous CMC with 5000 mPa s (Hvs-CMC group; 440.65 mPa s), respectively. The weight gain rate, specific growth rate, protein efficiency ratio, protein and lipid deposition rate in the CMC groups were dramatically lower than those in the control group, while feed conversion rate showed an opposite result. Plasma diamine oxidase activity, endothelin-1 and lipopolysaccharide concentrations in the Mvs-CMC and Hvs-CMC groups were significantly higher than in the control group, accompanied by a significant down-regulation of Occludin, Caludin-1 and Caludin-4. Intestinal glutathione concentration, superoxide dismutase and catalase activities in the CMC groups were significantly lower than in the control group, accompanied by a significant up-regulation of Keap1 and down-regulation of Nrf2. Moreover, CMC diets dramatically down-regulated the expression levels of IL-10 and TGF-β1. Digesta total short chain fatty acid and acetate concentrations in the CMC groups were dramatically higher than in the control group, while butyrate concentration showed an opposite result. The OTU, Sobs, Shannon and Simpson indices of intestinal flora in the CMC groups were dramatically lower than in the control group. Notably, structural analysis showed that dietary CMC dramatically increased the abundance of C. somerae and P. shigelloides , but reduced the abundance of C. colicanis and C. perfringens. In summary, increasing dietary viscosity adversely affects the intestinal flora structure and diversity, increases acetate/butyrate-producing bacterial ratio and the abundance of pathogenic microorganisms, disrupting intestinal flora homeostasis, impairs mucosa barrier function, induces intestinal inflammation and epithelial cell apoptosis in juvenile largemouth bass. Our findings demonstrate that soluble cellulose is more detrimental to intestinal health and growth in juvenile largemouth bass compared to insoluble cellulose, and the adverse effects of soluble cellulose are mainly caused by its viscosity. Importantly, this study demonstrate that viscosity is the main characteristic of non-starch polysaccharides that are detrimental to the intestinal health of fish. [ABSTRACT FROM AUTHOR]

The Affinity Grid technique combines sample purification and cryo-Electron Microscopy (cryo-EM) grid preparation into a single step. Several types of affinity surfaces, including functionalized lipids monolayers, streptavidin 2D crystals, and covalently functionalized carbon surfaces have been reported. More recently, we presented a new affinity cryo-EM approach, cryo-SPIEM, which applies the traditional Solid Phase Immune Electron Microscopy (SPIEM) technique to cryo-EM. This approach significantly simplifies the preparation of affinity grids and directly works with native macromolecular complexes without need of target modifications. With wide availability of high affinity and high specificity antibodies, the antibody-based affinity grid would enable cryo-EM studies of the native samples directly from cell cultures, targets of low abundance, and unstable or short-lived intermediate states. [ABSTRACT FROM AUTHOR]

Sleep-related hypermotor epilepsy (SHE) is a focal epilepsy syndrome characterized by seizures that predominantly occur during sleep. The pathogenesis of these seizures remains unclear. We previously detected rare variants in GABRG2 , which encodes the γ 2 subunit of γ-aminobutyric acid type A receptor (GABA A R), in patients with SHE and demonstrated that these variants impaired GABA A R function in vitro. However, the mechanisms by which GABRG2 variants contribute to seizure attacks during sleep remain unclear. In this study, we designed a knock-in (KI) mouse expressing the mouse Gabrg2 T316N variant, corresponding to human GABRG2 T317N variant, using CRISPR/Cas9. Continuous video-electroencephalogram monitoring and in vivo multichannel electrophysiological recordings were performed to explore seizure susceptibility to pentylenetetrazol (PTZ), alterations in the sleep-wake cycle, spontaneous seizure patterns, and synchronized activity in the motor thalamic nuclei (MoTN) and secondary motor cortex (M2). Circadian variations in the expression of total, membrane-bound, and synaptic GABA A R subunits were also investigated. No obvious changes in gross morphology were detected in Gabrg2 T316N/+ mice compared to their wild-type (Gabrg2 +/+ ) littermates. Gabrg2 T316N/+ mice share key phenotypes with patients, including sleep fragmentation and spontaneous seizures during sleep. Gabrg2 T316N/+ mice showed increased susceptibility to PTZ-induced seizures and higher mortality after seizures. Synchronization of the local field potentials between the MoTN and M2 was abnormally enhanced in Gabrg2 T316N/+ mice during light phase, when sleep dominates, accompanied by increased local activities in the MoTN and M2. Interestingly, in Gabrg2 +/+ mice, GABA A R γ2 subunits showed a circadian increase on the neuronal membrane and synaptosomes in the transition from dark phase to light phase, which was absent in Gabrg2 T316N/+ mice. We generated a new SHE mouse model and provided in vivo evidence that rare variants of GABRG2 contribute to seizure attacks during sleep in SHE. • Mice harboring the T316N variant in GABA A R γ 2 subunit are originally constructed. • Gabrg2 T316N/+ knock-in mice have spontaneous seizures mainly occurring during daytime. • NREM sleep is fragmented by brief awakening events in Gabrg2 T316N/+ knock-in mice. • Gabrg2 T316N/+ knock-in mice have enhanced synchronization between MoTN and M2. • Circadian change in membrane γ 2 subunit is absent in Gabrg2 T316N/+ knock-in mice. [ABSTRACT FROM AUTHOR]

[Display omitted] • In vitro digestion of MFGM protein and its relative stability and structural properties. • MFGM-derived peptides exhibit strong binding affinity with Keap1 protein. • Screening out two novel small-molecule antioxidant peptides and their effects on oxidative stress-injured myoblast. This study investigated the stability of milk fat globule membrane (MFGM) protein under simulated gastrointestinal conditions using an in vitro enzymatic digestion method. The optimal hydrolysis conditions were determined by monitoring the changes in particle size and zeta-potential of MFGM protein hydrolysates over time. Furthermore, the distribution of small molecular weight peptides with antioxidant activity was explored through DEAE-52 combined with in vitro cell experiments. Two novel antioxidant peptides (TGIIT and IITQ) were identified based on molecular docking technology and evaluated their potential scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2′-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS+) radicals. TGIIT and IITQ also demonstrated remarkable abilities in promoting mitochondrial biogenesis and activating Keap1/Nrf2 signaling pathway, which can effectively counteract skeletal muscle dysfunction induced by oxidative stress. Thus, MFGM-derived antioxidant peptides have the potential to be employed in food to regulate muscle protein metabolism and alleviate sarcopenia. [ABSTRACT FROM AUTHOR]

[Display omitted] • Co-fueling increases the current densities of C 3 -alcohols maximally by 9.9 times. • Product evolutions are recorded with in situ NMR monitoring. • Charge distributions are quantitatively analyzed with EC-NMR technique. • Co-fueling enhances the adsorption of –OH and the cleavage of C-C-C bonds. C 3 -alcohols can theoretically provide high current efficiencies in direct fuel cell applications because of their intrinsically large energy densities. However, during realistic reactions, molecular multiple carbon–carbon (C C C) bonds are hardly broken, severely reducing current efficiencies. Herein, directly with commercial Pt/C as the catalyst, the operation of co-fueling with formic acid significantly enhances the current efficiencies of two types of C 3 -alcohols, namely, isopropanol and 1,2-propanediol. Compared with the current densities of pure C 3 -alcohols, those of co-fueled C 3 -alcohols increase maximally by 9.9 times. The reason lies in the fact that the co-fueling design facilitates the breaking of C C C bonds, as elucidated by charge distributions that are obtained via electrochemical nuclear magnetic resonance analyses. Density-functional-theory computational results indicate that the generation of OH groups adsorbed on Pt surfaces is promoted in co-fueling experiments, reducing energy barriers for breaking progress. The co-fueling strategy is capable of optimizing C 3 -alcohol electrooxidations in direct fuel cells and deserves extensive study. [ABSTRACT FROM AUTHOR]

Abstract: Objective: We analyzed the clinical features and family-related gene mutations for the first two Chinese cases of type IV Waardenburg syndrome (WS4). Methods: Two families were analyzed in this study. The analysis included a medical history, clinical analysis, a hearing test and a physical examination. In addition, the EDNRB, EDN3 and SOX10 genes were sequenced in order to identify the pathogenic mutation responsible for the WS4 observed in these patients. Results: The two WS4 cases presented with high phenotypic variability. Two novel heterozygous mutations (c.254G>A and c.698-2A>T) in the SOX10 gene were detected. The mutations identified in the patients were not found in unaffected family members or in 200 unrelated control subjects. Conclusions: This is the first report of WS4 in Chinese patients. In addition, two novel mutations in SOX10 gene have been identified. [Copyright &y& Elsevier]

• Wideband (wide frequency range) acoustic modulation structures are designed. • The advantages of both periodic structures and porous materials are embodied here. • Theoretical and numerical modeling procedures are developed and validated. • Both the amplitude and bandwidth of sound transmission loss can be tuned. We proposed an effective acoustic abatement solution comprised of periodic resonators and multi-panel structures with porous lining, which incorporates the wideband capability of porous materials and the low-frequency advantage of locally resonant structures together. Theoretical model and numerical implementation are developed and validated. Two-dimensional poroelastic field expressions are used and resonator forces are incorporated. The results agree well with those reported in the literature and the results obtained from the finite element method. It turns out that sound insulation concerning both the amplitude and tuning bandwidth can be achieved effectively using porous additions and locally-resonant designs. This study presents a promising and practical alternative for wideband acoustic modulation. [ABSTRACT FROM AUTHOR]

The emergence of worldwide spreading drug-resistant bacteria has been a serious threat to public health during the past decades. The development of new and effective antibacterial agents to address this critical issue is an urgent action. In the present study, we investigated the antibacterial activity of two 9,10-dihydroacridine derivatives and their mechanism. Both compounds were found possessing strong antibacterial activity against some selected Gram-positive bacteria including MRSA, VISA and VRE. The biological study suggests that the compounds promoted FtsZ polymerization and also disrupted Z-ring formation at the dividing site and consequently, the bacterial cell division is interrupted and causing cell death. Image 1 • B1 and B2 possess strong antibacterial activity against Gram-positive bacteria. • B1 and B2 disrupted FtsZ function by inducing FtsZ polymerization. • B1 and B2 can restore antibacterial activity of methicillin against MRSA. [ABSTRACT FROM AUTHOR]

Abstract: Diabetes mellitus (DM) is a serious metabolic disorder with micro- and macrovascular complications that results in significant morbidity and mortality. It is well established that cytosolic Ca2+ play an important role in controlling insulin secretion in pancreatic β-cells. The Na+/Ca2+ exchanger (NCX), an ion transport protein, is expressed in the plasma membrane of virtually all animal cells. NCX is a reversible carrier that can mediate the transport of Ca2+ across the plasma membrane in both directions. Therefore, great efforts have been made to identify NCX associated with DM. NCX is expressed in several tissues, and acts in the protection against intracellular calcium overload; in the regulation of insulin secretion by beta cells, and in improving vascular endothelium-dependent relaxation. All these mechanisms are associated with DM pathogenesis and its chronic complications. Therefore, NCX is a candidate protein for the development of these disorders. Only a few studies investigated NCX in relation to chronic complications of diabetes, with inconclusive results. [Copyright &y& Elsevier]

Avian colibacillosis is one of the most serious infectious bacterial diseases that endanger the modern poultry industry. Lactobacillus is believed to inhibit intestinal pathogens and maintain a healthy gut microbiota. This study aimed to investigate Lactobacillus supplementation in Cherry Valley ducks to prevent the intestinal flora dysbiosis caused by Duck Escherichia coli 17. One hundred and twenty healthy one day old Cherry Valley ducks were randomized to three study groups (Group I = the control group; Group II = duck Escherichia coli 17 challenge group and Group III = DE17 challenge group supplemented with lactic acid bacteria composite preparation). Cherry Valley ducks in Group II and Group III were gavage challenged with DE17 (1 × 105 CFU/mL) on day 14. Pyrosequencing of the V3/V4 variable regions of the genes encoding for 16S rRNA was used for sequence analysis. The results showed that the normal intestinal microecology was affected by DE17, including a relative increase in proteobacteria. At the same time, the Lactobacillales were increased and harmful bacteria were decreased in different intestinal segments of ducks in Group III, compared to those in Group II. Network analysis showed that dietary lactic acid bacteria addition improved the interaction pattern within the cecal microbiota of ducks and the result showed that in Ruminococcus _2 was independently present in the group III and Lachnospiraceae _NK4A136_group species correlation existed between group I and group III. This study proved that oral supplementation with Lactobacillus casei 1.2435, Lactobacillus rhamnosus 621 and Lactobacillus rhamnosus A4 can mitigate DE17 induced intestinal flora dysbiosis. • Lactobacillus with good probiotic potential were isolated from Cherry Valley Ducks. • Pre-supplementation of Lactobacillus effectively alleviate mitigate by Duck Escherichia coli 17 induced intestinal flora dysbiosis. • Lactobacillus composite preparation supplementation increased the population of Lactobacillus in the cecal. [ABSTRACT FROM AUTHOR]

Phage G has the largest capsid and genome of any known propagated phage. Many aspects of its structure, assembly, and replication have not been elucidated. Herein, we present the dsDNA-packed and empty phage G capsid at 6.1 and 9 Å resolution, respectively, using cryo-EM for structure determination and mass spectrometry for protein identification. The major capsid protein, gp27, is identified and found to share the HK97-fold universally conserved in all previously solved dsDNA phages. Trimers of the decoration protein, gp26, sit on the 3-fold axes and are thought to enhance the interactions of the hexameric capsomeres of gp27, for other phages encoding decoration proteins. Phage G's decoration protein is longer than what has been reported in other phages, and we suspect the extra interaction surface area helps stabilize the capsid. We identified several additional capsid proteins, including a candidate for the prohead protease responsible for processing gp27. Furthermore, cryo-EM reveals a range of partially full, condensed DNA densities that appear to have no contact with capsid shell. Three analyses confirm that the phage G host is a Lysinibacillus , and not Bacillus megaterium : identity of host proteins in our mass spectrometry analyses, genome sequence of the phage G host, and host range of phage G. Unlabelled Image • Cryo-EM reveals structure of phage G capsid at 6.1 Å resolution. • Interactions of the phage G decoration protein trimer with the major capsid protein shell resemble those of Lambda and TW1 phages. • Cryo-EM micrographs of phage G reveal condensed, smaller-than-capsid DNA densities. • Mass spectrometry analysis of phage G identifies (1) all major capsid components and (2) the prohead protease responsible for the processing of the major capsid protein, gp27. • Phage G structural studies led to proteomic and genomic analyses that revised the identity of the phage G host from Bacillus megaterium to a Lysinibacillus species. [ABSTRACT FROM AUTHOR]

• MBIR can significantly boost contrast of tomograms of cryo specimens. • MBIR can significantly reduce missing wedge artifacts and retain sharp features. • MBIR can partially restore the missing information in the unsampled angular region. • MBIR can improve the resolution of subtomogram averages. Cryo-Electron Tomography (cryo-ET) has become an essential technique in revealing cellular and macromolecular assembly structures in their native states. However, due to radiation damage and the limited tilt range, cryo-ET suffers from low contrast and missing wedge artifacts, which limits the tomograms to low resolution and hinders further biological interpretation. In this study, we applied the Model-Based Iterative Reconstruction (MBIR) method to obtain tomographic 3D reconstructions of experimental cryo-ET datasets and demonstrated the advantages of MBIR in contrast improvement, missing wedge artifacts reduction, missing information restoration, and subtomogram averaging compared with other reconstruction approaches. Considering the outstanding reconstruction quality, MBIR has a great potential in the determination of high resolution biological structures with cryo-ET. [ABSTRACT FROM AUTHOR]

Volta Phase Plate (VPP) has become an invaluable tool for cryo-EM structural determination of small protein complexes by increasing image contrast. Currently, the standard protocol of VPP usage periodically changes the VPP position to a fresh spot during data collection. Such a protocol was to target the phase shifts to a relatively narrow range (around 90°) based on the observations of increased phase shifts and image blur associated with more images taken with a single VPP position. Here, we report a 2.87 Å resolution structure of apoferritin reconstructed from a dataset collected using only a single position of VPP. The reconstruction resolution and map density features are nearly identical to the reconstruction from the control dataset collected with periodic change of VPP positions. Further experiments have verified that similar results, including a 2.5 Å resolution structure, could be obtained with a full range of phase shifts, different spots of variable phase shift increasing rates, and at different ages of the VPP post-installation. Furthermore, we have found that the phase shifts at low resolutions, probably related to the finite size of the Volta spots, could not be correctly modeled by current CTF model using a constant phase shift at all frequencies. In dataset III, severe beam tilt issue was identified but could be computationally corrected with iterative refinements. The observations in this study may provide new insights into further improvement of both the efficiency and robustness of VPP, and to help turn VPP into a plug-and-play device for high-resolution cryo-EM. [ABSTRACT FROM AUTHOR]

Anisotropic magnification distortion of TEM images (mainly the elliptic distortion) has been recently found as a potential resolution-limiting factor in single particle 3-D reconstruction. Elliptic distortions of ∼1–3% have been reported for multiple microscopes under low magnification settings (e.g., 18,000×), which significantly limited the achievable resolution of single particle 3-D reconstruction, especially for large particles. Here we report a generic algorithm that formulates the distortion correction problem as a generalized 2-D alignment task and estimates the distortion parameters directly from the particle images. Unlike the present pre-calibration methods, our computational method is applicable to all datasets collected at a broad range of magnifications using any microscope without need of additional experimental measurements. Moreover, the per-micrograph and/or per-particle level elliptic distortion estimation in our method could resolve potential distortion variations within a cryo-EM dataset, and further improve the 3-D reconstructions relative to constant-value correction by the pre-calibration methods. With successful applications to multiple datasets and cross-validation with the pre-calibration method, we have demonstrated the validity and robustness of our algorithm in estimating the distortion; correction of the elliptic distortion significantly improved the achievable resolutions by ∼1–3 folds and enabled 3-D reconstructions of multiple viral structures at 2.4–2.6 Å resolutions. The resolution limits with elliptic distortion and the amounts of resolution improvements with distortion correction were found to strongly correlate with the product of the particle size and the amount of distortion, which can help assess if elliptic distortion is a major resolution limiting factor for single particle cryo-EM projects. [ABSTRACT FROM AUTHOR]