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Summary: Clinical research indicates that SARS-CoV-2 infection is linked to several neurological consequences, and the virus is still spreading despite the availability of vaccinations and antiviral medications. To determine how hosts respond to SARS-CoV-2 infection, we employed LC-MS/MS to perform ubiquitinome and proteome analyses of the brain cortexes from K18-hACE2 mice in the presence and absence of SARS-CoV-2 infection. A total of 8,024 quantifiable proteins and 5,220 quantifiable lysine ubiquitination (Kub) sites in 2023 proteins were found. Glutamatergic synapse, calcium signaling pathway, and long-term potentiation may all play roles in the neurological consequences of SARS-CoV-2 infection. Then, we observed possible interactions between 26 SARS-CoV-2 proteins/E3 ubiquitin-protein ligases/deubiquitinases and several differentially expressed mouse proteins or Kub sites. We present the first description of the brain cortex ubiquitinome in K18-hACE2 mice, laying the groundwork for further investigation into the pathogenic processes and treatment options for neurological dysfunction following SARS-CoV-2 infection.

The estrogen receptor alpha (ERα) plays a central role in the etiology, progression, and treatment of breast cancers. Constitutively activating somatic mutations Y537S and D538G, in the ligand binding domain (LBD) of ESR1, are associated with acquired resistance to endocrine therapies. We have previously shown that the metalloestrogen calcium activates ERα through an interaction with the LBD of the receptor. This study shows that cadmium activates ERα through a mechanism similar to calcium and contributes to, and further increases, the constitutive activity of the ERα mutants Y537S and D538G. Mutational analysis identified C381, N532A, H516A/N519A/E523A, and E542/D545A on the solvent accessible surface of the LBD as possible calcium/metal interaction sites. In contrast to estradiol, which did not increase the activity of the Y537S and D538G mutants, cadmium increased the activity of the constitutive mutants. Mutation of the calcium/metal interaction sites in Y537S and D538G mutants resulted in a significant decrease in constitutive activity and cadmium induced activity. Mutation of calcium/metal interaction sites in wtERα diminished binding of the receptor to the enhancer of estrogen responsive genes and the binding of nuclear receptor coactivator 1 and RNA polymerase II. In contrast to wtERα, mutation of the calcium/metal interaction sites in the Y537S and D538G mutants did not diminish binding to DNA but prevented a stable interaction with the coactivator and polymerase. Growth assays further revealed that calcium channel blockers and chelators significantly decreased the growth of MCF7 cells expressing these constitutively active mutants. Taken together, the results suggest that exposure to cadmium plays a role in the etiology, progression, and response to treatment of breast cancer due, in part, to its ability to activate ERα.

The abundance of petroleum resource is influenced by various factors and involves complex parameters and extensive data. Consequently, traditional geostatistical methods often lack precision in quantitative prediction. To address this issue, this study focuses on the third member of Paleogene Shahejie Formation (member Es3) in the Wenliu area of the Dongpu Sag and utilizes a multi-layer perceptron neural network (MLP) for predicting petroleum resource abundance and employed the Boosting ensemble learning algorithm to optimize the prediction model. The MLP and MLP-Boosting algorithm models were test on 66 sample groups, yielding correlation coefficients of 0.789 and 0.989 for the training set, 0.618 and 0.825 for the validation set and 0.689 and 0.845 for the test set. The analysis identified effective thickness, average permeability and effective porosity are the most significant geological factors influencing petroleum resource abundance, with importance coefficients of 33.93%, 20.12% and 19.53%, respectively. Other factors such as trap area, surface crude oil density and sedimentary facies assignment were found to be less influential. Overall, the Boosting ensemble learning algorithm significantly enhanced the prediction accuracy of the multi-layer perceptron model, offering valuable insights for target optimization, exploration planning and petroleum resource evaluation in other blocks in the sag.

Understanding aeroelastic issues related to isolators is pivotal for the structural design and flow control of scramjets. However, research on fluid–structure interactions (FSIs) between thin-walled structures and the isolator flow remains limited. This study delves into the FSIs between thin-walled panels and the isolator flow, as characterized by an oblique shock train, by quantitatively analyzing 11 flow parameters assessing the structural response, separation zones, shock structures, flow symmetry, and performance. The results reveal that an FSI triggers panel flutter under oblique shock train conditions, with the panel shapes exhibiting a combination of first- and second-mode responses, peaking at 0.75 of the panel length. Compared to rigid wall conditions, isolators with a flexible panel at the bottom wall experience downstream movement of the separation zones and shock structures, reduced flow symmetry, and minor changes in performance. Transient fluctuations occur due to the panel flutter. Two flexible panels at the top and bottom walls have a comparatively lesser influence on the averaged parameters but exhibit more violent transient fluctuations. Furthermore, the FSI effects under oblique shock train conditions are contrasted with those under normal shock train conditions. The flutter response under normal shock train conditions is more pronounced, with a larger amplitude and higher frequency, driven by the heightened participation of the first-mode response. The effects of FSIs under normal shock train conditions on the averaged parameters are the opposite (with a larger influence) to those under oblique shock train conditions, with significantly more drastic transient fluctuations. Overall, this study sheds light on the complex and substantial influence of FSIs on the isolator flow, emphasizing the necessity of considering FSIs in future isolator design and development endeavors.

Como imagen fundamental de la poesía clásica china, el agua cobra una notable importancia en la obra de Su Shi (Su Dongpo). En este trabajo se analizan los significados de este elemento natural en las composiciones de Su Shi relacionadas con la nostalgia por la tierra natal —uno de los temas más representativos de su creación poética— según las diferentes distancias psicológicas entre el autor y la imagen, las cuales podrían clasificarse en tres: la evocación de la nostalgia por tierra natal, la personificación y la espiritualización. A través de la traducción y las observaciones analíticas, se estudia cómo el poeta asimila la tradición literaria para después innovar respecto a la simbología del agua. El artículo desvela las influencias de las doctrinas confucianistas y daoístas en el autor, lo que ayuda a comprender su mundo interior manera más profunda.

TPC2 is an important organellar Na+/Ca2+ release channel which regulates function of lysosomes and lysosome-related organelles. Here, Wang et al. demonstrate that a gain-of-function mutation (R210C) in TPC2 leads to hypopigmentaion, enlarged endolysosomes, enhanced lysosomal Ca2+ release and hyper-acidification.

Regulated secretion is conserved in all eukaryotes. In vertebrates granin family proteins function in all key steps of regulated secretion. Phase separation and amyloid-based storage of proteins and small molecules in secretory granules require ion homeostasis to maintain their steady states, and thus need ion conductances in granule membranes. But granular ion channels are still elusive. Here we show that granule exocytosis in neuroendocrine cells delivers to cell surface dominant anion channels, to which chromogranin B (CHGB) is critical. Biochemical fractionation shows that native CHGB distributes nearly equally in soluble and membrane-bound forms, and both reconstitute highly selective anion channels in membrane. Confocal imaging resolves granular membrane components including proton pumps and CHGB in puncta on the cell surface after stimulated exocytosis. High pressure freezing immuno-EM reveals a major fraction of CHGB at granule membranes in rat pancreatic β-cells. A cryo-EM structure of bCHGB dimer of a nominal 3.5 Å resolution delineates a central pore with end openings, physically sufficient for membrane-spanning and large single channel conductance. Together our data support that CHGB-containing (CHGB+) channels are characteristic of regulated secretion, and function in granule ion homeostasis near the plasma membrane or possibly in other intracellular processes.

IntroductionRealgar has a long history ofuse in traditional medicines. However, the mechanism through which Realgar or Realgar-Indigo naturalis formula (RIF) exert therapeutic effects is only partially understood.MethodsIn this study, 60 feces and 60 ileum samples from rats administered with realgar or RIF were collected to examine the gut microbiota.ResultsThe results showed that realgar and RIF influenced different microbiota in both feces and ileum. Compared with realgar, RIF at low dosage (0.1701 g/3 ml) significantly increased the microbiota diversity. LEfSe and random forest analyses showed that the bacterium Bacteroidales was significantly altered after RIF administration, and it was predicted that these microorganisms contribute to the inorganic arsenic metabolic process.DiscussionOur results suggest that realgar and RIF may exert their therapeutic effects through influencing microbiota. The low dose of RIF had greater effects on increasing the diversity of microbiota, and Bacteroidales in feces might participate in the inorganic arsenic metabolic process to exert therapeutic effects for realgar.

This study aims to determine the sources of indoor particulate matter at the Potala Palace Museum in Tibet, China, and evaluate the potential hazards of these pollutants for cultural relics. Long-term monitoring of indoor and outdoor suspended particulate matter concentrations was conducted, and sediment particle samples were collected. The chemical composition of the deposited particles was determined using X-ray fluorescence analysis (XRF). The outdoor suspended particulate concentration was much lower than that indoors; the indoor PM1-10 concentration was much higher than that outdoors and was less affected by outdoor sources. The sources of indoor deposited particles in the high-plateau museum can be classified into four categories: soil dust brought in by tourists from the outdoors, incense ash, pollution from human activities, and ores. Based on data analysis and discussion, proper ventilation can dilute indoor suspended particulate matter, and the installation of air conditioning systems can control temperature and humidity at 20 °C and about 45–60%, respectively, and reduce the fluctuation value, so as to promote particle deposition and better protect the museum’s cultural relics.

The unique climate and the landscape of severely cold regions in winter attract many tourists. The outdoor thermal environment affects the space use and the tourist experience, becoming one of the key factors in the design of tourist attractions. The outdoor thermal comfort of tourists from different regions should be considered, but it has been poorly studied in winter in severely cold regions. This paper explores the differences in outdoor thermal comfort in winter between local and non-local tourists through the field measurement of the thermal environment and a questionnaire survey of thermal comfort at tourist attractions in Harbin, China. The results show that the proportion of local tourists who expect the air temperature and solar radiation to rise in winter is higher than that of non-local tourists. The thermal sensation vote of local tourists is generally higher than that of non-local tourists. When the Physiologically Equivalent Temperature (PET) < −6 °C, the thermal satisfaction of non-local tourists is higher than that of local tourists. When the PET value is −10 °C, the thermal comfort of non-local tourists is the highest. The thermal comfort decreases with the rise or fall of the PET value. When −28 °C < PET < −7 °C, the thermal comfort of non-local tourists is generally higher than that of local tourists. This paper provides a reference and evaluation basis for urban tourist attractions’ outdoor thermal environment design in severely cold regions.

Background. Although prostate cancer patients initially respond to androgen deprivation therapy, most patients progress to a resistant phenotype. Castration resistance is due, in part, to intratumoral and/or adrenal synthesis of androgens, overexpression or mutation of the androgen receptor (AR), stabilization of AR by chaperones, and ligand-independent activation of AR. Increasing evidence also links disruption of calcium homeostasis to progression of prostate cancer. Our previous study shows that heavy metal cadmium activates the AR through a ligand-independent mechanism. Cadmium mimics calcium in biological systems due to their similar ionic charge and radius. This study determines whether calcium activates AR and whether first- and second-generation antiandrogens block the ability of calcium to activate the receptor. Methods. The expression of androgen-responsive genes and calcium channels was measured in prostate cells using a quantitative real-time polymerase chain reaction assay. Cell growth was measured. Results. To ask whether calcium activates AR, prostate cells were treated with calcium in the absence and presence of the first-generation antiandrogens hydroxyflutamide and bicalutamide and the second-generation antiandrogen enzalutamide, and the expression of androgen-responsive genes and cell growth was measured. In the normal PWR-1E cells and HEK293T cells transiently expressing AR, treatment with calcium increased the expression of androgen-responsive genes by approximately 3-fold. The increase was blocked by enzalutamide but was not consistently blocked by the first-generation antiandrogens. In LNCaP cells which contain a mutant AR, treatment with calcium also increased the expression of androgen-responsive genes by approximately 3-fold, and the increase was more effectively blocked by enzalutamide than by hydroxyflutamide or bicalutamide. Treatment with calcium also increased cell growth that was blocked by enzalutamide. To ask whether dysregulation of calcium channels is associated with castration resistance, calcium channels were measured in the normal PWR-1E prostate cells, the hormone-responsive LNCaP cells, and the castration-resistant VCaP and 22RV1 cells. Compared to normal prostate cells, the hormone-responsive and hormone-resistant cells overexpressed several calcium channels. Conclusions. The results of this study show that calcium activates AR and increases cell growth and that calcium channels are overexpressed in hormone-responsive and hormone-resistant prostate cancer cells. Taken together, the results suggest a novel role of calcium in the castration-resistant phenotype.

Abstract Background Cisplatin is a platinum-based chemotherapeutic that damages genomic DNA leading to cell death. It also damages mitochondrial DNA and induces high levels of mitochondrial reactive oxygen species (mtROS), further sensitising cells to apoptosis. Notably, immature oocytes are particularly vulnerable to cisplatin treatment, a common side effect of which is depletion of the primordial follicle reserve, leading to infertility and early menopause. Cisplatin is known to damage the DNA of oocytes, but the possibility that cisplatin also compromises oocyte survival and quality by damaging mitochondria, has not been investigated. To begin to address this question, neonatal mice were treated with saline or cisplatin (2 mg/kg or 4 mg/kg) and the short and long-term impacts on mitochondria in oocytes were characterised. Results At 6 and 24 h after treatment, mitochondrial localisation, mass and ATP content in immature oocytes were similar between groups. However, TMRM staining intensity, a marker of mitochondrial membrane potential, was decreased in immature oocytes from cisplatin treated mice compared to saline treated controls, consistent with the induction of apoptosis. When mice were super ovulated 5 weeks after exposure, the number of mature oocytes harvested from cisplatin treated mice was significantly lower than controls. Mitochondrial localisation, mass, membrane potential and ATP levels showed no differences between groups. Conclusions These findings suggest that mitochondrial dysfunction may contribute to the depletion of the ovarian reserve caused by cisplatin, but long-term impacts on mitochondria may be minimal as those immature oocytes that survive cisplatin treatment develop into mature oocytes with normal mitochondrial parameters.

Misophonia involves a decreased tolerance to certain sounds and is associated with a range of emotions and emotion processes. In addition to the distress caused by misophonia, some individuals report having aggressive outbursts and significant impact on doing things they would like to be able to do. This study aimed to examine whether misophonia-specific cognitive and emotional processes were associated with misophonic outbursts and impact, and whether these relationships could be explained in part by emotion processes not specific to misophonia. A sample of 703 individuals, 315 of whom identified with having misophonia, completed measures of misophonia, depression and anxiety symptoms, anxiety and disgust sensitivity, interoception and beliefs about emotions. Exploratory correlation and regression analyses were used to build mediation models, which were tested using multiple linear regression. Externalising appraisals (blaming others for causing one’s reaction to sounds) were positively associated with misophonic outbursts, and this relationship was partially explained by anxiety symptoms and disgust sensitivity. Sense of emotional threat in misophonia predicted functional impact of misophonia, and this was partially explained by depression symptoms and negative beliefs about emotions. Anxiety sensitivity and interoception were not significant independent predictors of misophonic outbursts or functional impact. These results provide support for the relevance of emotion processes in misophonia and highlight the importance of using multi-dimensional measures of misophonia to improve our understanding of the condition.

Misophonia is a disorder generally characterised by a decreased tolerance to everyday sounds. Although research is increasing in misophonia, a cross-cultural validation of a psychometric tool for measuring misophonia has not been evaluated. This study investigated the validity of the S-Five multidimensional model of the misophonic experience in a sample of Chinese participants. The S-Five was translated in a forward-backward method to Mandarin to establish a satisfactory translation. The translation was also independently back translated to English, with no significant differences when compared to the original S-Five. Through exploratory factor analysis, using responses from 256 Chinese individuals, the five dimensions (internalising appraisals, externalising appraisals, perceived threat and avoidance behaviour, outbursts, and impact on functioning) were replicated, indicating the cross-cultural uniformity of the experience of misophonia as captured by the S-Five. That is, current results point to the stability of the manifestation of misophonia across cultures, seen here for the first time in the literature. By design, the S-Five items were developed to reflect sound sensitivities in a manner that is not specific or matching to individuals of a certain age, gender, ethnicity, nationality, socio-economic status, and educational level. Testimonial to this fact is not only the replication of the five factors, but also the replication of the evidence towards satisfactory psychometric properties (reliability and validity) of the scale. Based on the results of this study, the S-Five is a psychometrically robust tool to be used within the Chinese population.

Agarwood, a highly valuable resin/wood combination with diverse pharmacological activities but scarce supply, has a long history of being used as a medicine in several medical systems. Grafted Kynam agarwood (GKA) has been cultivated successfully recently and has the qualities meeting the definition of premium Kynam agarwood. However, there are few comprehensive comparisons between GKA and normal agarwood in terms of traits, global composition, and activity, and some key issues for GKA to be adopted into the traditional Chinese medical (TCM) system have not been elaborated. The two types of agarwood samples were evaluated in terms of trait characteristics, physicochemical indicators, key component groups, and global compositional profile. Furthermore, a molecular docking was performed to investigate the active ingredients. In vitro activity assays were performed to evaluate the activation of adenosine 5’-monophosphate (AMP)-activated protein kinase (AMPK) by GKA and normal agarwood. The results revealed that, overall, the traits, microscopic characteristics, chemical composition types, and bioactivity between GKA and normal agarwood were similar. The main differences were the content of resin (ethanolic extract content), the content of key component groups, and the composition of the different parent structural groups of 2-(2-phenethyl) chromones (PECs). The contents of total PEC and ethanol extract content of GKA were significantly higher than those of normal agarwood. The MS-based high-throughput analysis revealed that GKA has higher concentrations of sesquiterpenes and flindersia-type 2-(2-phenylethyl) chromones (FTPECs) (m/z 250-312) than normal agarwood. Molecular docking revealed that parent structural groups of FTPECs activated multiple signaling pathways, including the AMPK pathway, suggesting that FTPECs are major active components in GKA. The aim of this paper is to describe the intrinsic reasons for GKA as a high-quality agarwood and a potential source for novel drug development. We combined high-throughput mass spectrometry and multivariate statistical analysis to infer the different components of the two types of agarwood. Then we combined virtual screening and in vitro activity to construct a component/pharmacodynamic relationship to explore the causes of the activity differences between agarwood with different levels of quality and to identify potentially valuable lead compounds. This strategy can also be used for the comprehensive study of other TCMs with different qualities.

The Lufeng Sag and Huizhou Sag, both located in the Zhu-1 Depression, have similar geographical locations, but their reservoir characteristics in the Paleogene Wenchang Formation show obvious differences. Primary intergranular pores are mainly developed in the Lufeng Sag. However, secondary pores are the main reservoir space in the Huizhou depression. Overall, the reservoir properties of the Lufeng Sag are better than those of the Huizhou Sag. To analyse the differences between the Paleogene reservoirs in these two areas, this study mainly uses assay data, such as rock thin sections, scanning electron microscope images, drilling, and logging, to analyse the differential development mechanisms of high-quality reservoirs, and two types of reservoir development models were concluded. The results show that the anti-compaction primary porosity preservation mode is mainly developed in the Lufeng Sag. High compositional maturity quartz sandstone is the congenital condition of primary porosity development. The top and bottom calcareous cementation formed of the large set of thick sand bodies increases the rock’s anti-compaction ability. The early shallow burial slows down the compaction action of overlying strata. Under the low geothermal temperature, it can delay the time for deep reservoirs to enter the middle diagenetic stage. The reservoirs in the Huizhou Sag are developed with the secondary dissolution pore development model. The Wenchang Formation reservoir in the Huizhou Sag has a large area of contact with source rocks, and organic acids can migrate to sandstone reservoirs for dissolution. Additionally, the secondary dissolution pores are more developed because the Wenchang Formation reservoirs in the Huizhou Sag contain more easily dissolved substances.

AMS2, amulticopy suppressor for the cpn1 (SpCENP-A) mutant, functions to specifically regulate histone genes transcription and chromosome segregation. As a cell-cycle-regulated GATA transcription factor in eukaryotic organisms, little research has been done on the role of AMS2 protein in pathogenic fungi. In Sclerotinia sclerotiorum, Ssams2 (SS1G_03252) encodes a protein which has been predicted to contain GATA-box domain. Here, Ssams2-silenced strains with significantly reduced Ssams2 gene expression levels exhibited defect in hyphal growth, hyphal branching patterns, compound appressoria differentiation and the oxalic acid production compared to the wild-type (WT) strain. By common bean leaves infection assays, we identified the role of Ssams2 in full virulence. Furthermore, the numbers of cell nucleus in the same length of mycelium in Ssams2-silenced transformants were significantly less than that in the WT strain. The expression levels of histone genes and cell cycle genes in transformants were down-regulated significantly in the RNAi strains. Taken together, our work suggests that the TF SsAMS2 is required for growth, appressoria formation, virulence, and chromosome segregation in S. sclerotiorum.

One important factor for successful disease management is the ability to rapidly and accurately identify the causal agent. Plant viruses cause severe economic losses and pose a serious threat to sustainable agriculture. Therefore, optimization of the speed, sensitivity, feasibility, portability, and accuracy of virus detection is urgently needed. Here, we developed a clustered regularly interspaced short palindromic repeats (CRISPR)-based nucleic acid diagnostic method utilizing the CRISPR–Cas12a system for detecting two geminiviruses, tomato yellow leaf curl virus (TYLCV) and tomato leaf curl New Delhi virus (ToLCNDV), which have single-stranded DNA genomes. Our assay detected TYLCV and ToLCNDV in infected plants with high sensitivity and specificity. Our newly developed assay can be performed in ~1 h and provides easy-to-interpret visual readouts using a simple, low-cost fluorescence visualizer, making it suitable for point-of-use applications.

SFH1 (for Snf5 homolog) protein, comprised in the RSC (Remodels Structure of Chromatin) chromatin remodeling complex, functions as a transcription factor (TF) to specifically regulate gene transcription and chromatin remodeling. As one of the well-conserved TFs in eukaryotic organisms, little is known about the roles of SFH1 protein in the filamentous fungi. In Sclerotinia sclerotiorum, one of the notorious plant fungal pathogens, there are nine proteins predicted to contain GATA-box domain according to GATA family TF classification, among which Sssfh1 (SS1G_01151) encodes a protein including a GATA-box domain and a SNF5 domain. Here, we characterized the roles of Sssfh1 in the developmental process and fungal pathogenicity by using RNA interference (RNAi)-based gene silencing in S. sclerotiorum. RNA-silenced strains with significantly reduced Sssfh1 RNA levels exhibited slower hyphal growth and decreased reactive oxygen species (ROS) accumulation in hyphae compared to the wild-type (WT) strain. Yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) assays demonstrated that SsSFH1 interacts with SsMSG5, a MAPK phosphatase in S. sclerotiorum. Furthermore, Sssfh1-silenced strains exhibited enhanced tolerance to NaCl and H2O2. Results of infection assays on soybean and common bean (Phaseolus vulgaris) leaves indicated that Sssfh1 is required for full virulence of S. sclerotiorum during infection in the susceptible host plants. Collectively, our results suggest that the TF SsSFH1 is involved in growth, ROS accumulation and virulence in S. sclerotiorum.

A computer virus model with infection delay and recovery delay is considered. The sufficient conditions for the global stability of the virus infection equilibrium are established. We show that the time delay can destabilize the virus infection equilibrium and give rise to Hopf bifurcations and stable periodic orbits. By the normal form and center manifold theory, the direction of the Hopf bifurcation and stability of the bifurcating periodic orbits are determined. Numerical simulations are provided to support our theoretical conclusions.

Site-specific nucleases are crucial for genome engineering applications in medicine and agriculture. The ideal site-specific nucleases are easily reprogrammable, highly specific in target site recognition, and robust in nuclease activities. Prokaryotic Argonaute (pAgo) proteins have received much attention as biotechnological tools due to their ability to recognize specific target sequences without a protospacer adjacent motif, but their lack of intrinsic dsDNA unwinding activity limits their utility in key applications such as gene editing. Recently, we developed a pAgo-based system for site-specific DNA cleavage at physiological temperatures independently of the DNA form, using peptide nucleic acids (PNAs) to facilitate unwinding dsDNA targets. Here, we fused catalytically dead pAgos with the nuclease domain of the restriction endonuclease FokI and named this modified platform PNA-assisted FokI-(d)pAgo (PNFP) editors. In the PNFP system, catalytically inactive pAgo recognizes and binds to a specific target DNA sequence based on a programmable guide DNA sequence; upon binding to the target site, the FokI domains dimerize and introduce precise dsDNA breaks. We explored key parameters of the PNFP system including the requirements of PNA and guide DNAs, the specificity of PNA and guide DNA on target cleavage, the optimal concentration of different components, reaction time for invasion and cleavage, and ideal temperature and reaction buffer, to ensure efficient DNA editing in vitro. The results demonstrated robust site-specific target cleavage by PNFP system at optimal conditions in vitro. We envision that the PNFP system will provide higher editing efficiency and specificity with fewer off-target effects in vivo. [ABSTRACT FROM AUTHOR]

The estrogen receptor alpha (ERa) plays a central role in the etiology, progression, and treatment of breast cancers. Constitutively activating somatic mutations Y537S and D538G, in the ligand binding domain (LBD) of ESR1, are associated with acquired resistance to endocrine therapies. We have previously shown that the metalloestrogen calcium activates ERa through an interaction with the LBD of the receptor. This study shows that cadmium activates ERa through a mechanism similar to calcium and contributes to, and further increases, the constitutive activity of the ERa mutants Y537S and D538G. Mutational analysis identified C381, N532A, H516A/N519A/E523A, and E542/ D545A on the solvent accessible surface of the LBD as possible calcium/metal interaction sites. In contrast to estradiol, which did not increase the activity of the Y537S and D538G mutants, cadmium increased the activity of the constitutive mutants. Mutation of the calcium/metal interaction sites in Y537S and D538G mutants resulted in a significant decrease in constitutive activity and cadmium induced activity. Mutation of calcium/metal interaction sites in wtERa diminished binding of the receptor to the enhancer of estrogen responsive genes and the binding of nuclear receptor coactivator 1 and RNA polymerase II. In contrast to wtERa, mutation of the calcium/metal interaction sites in the Y537S and D538G mutants did not diminish binding to DNA but prevented a stable interaction with the coactivator and polymerase. Growth assays further revealed that calcium channel blockers and chelators significantly decreased the growth of MCF7 cells expressing these constitutively active mutants. Taken together, the results suggest that exposure to cadmium plays a role in the etiology, progression, and response to treatment of breast cancer due, in part, to its ability to activate ERa. [ABSTRACT FROM AUTHOR]

With the rapid renewal and development of metro lines, the activity disturbances caused by metro noise inside buildings have attracted much attention. A large number of studies have explored the activity disturbances induced by aboveground railway or road traffic noises, there is a lack of research assessing the activity disturbances due to metro noise. Accordingly, this study investigated the disturbances induced by metro noise in indoor condition at four noise levels (50, 55, 60 and 65 dBA) in three kinds of stimulation conditions (calculation, judgement andmemory) and the regulating effects of adding four types ofwater sounds (fountain, stream,water-drop andwaterfall sounds) on the disturbance levels due to metro noise. Seventy-nine subjects participated in the experiment and were exposed to a series of acoustic samples before being asked to evaluate the disturbance levels using a 5-point verbal scale. The results show that under the same metro noise environments, the disturbance levels of memory weremuch higher than those of calculation and judgement, and compared to the disturbance level of calculation, the disturbance levels of judgement and memory were more sensitive to the change of metro noise level. Besides, when global A-weighted equivalent sound pressure level lay between 55 and 65 dBA, the disturbance level of memory due tometro noise seemed to reduce after adding the fountain sound orwaterdrop sound, among which the capability of water-drop sound seemed stronger, and the mitigating effect of water-drop sound seemed to become more significant with the increase of global LAeq. [ABSTRACT FROM AUTHOR]

Regulated secretion is conserved in all eukaryotes. In vertebrates granin family proteins function in all key steps of regulated secretion. Phase separation and amyloid-based storage of proteins and small molecules in secretory granules require ion homeostasis to maintain their steady states, and thus need ion conductances in granule membranes. But granular ion channels are still elusive. Here we show that granule exocytosis in neuroendocrine cells delivers to cell surface dominant anion channels, to which chromogranin B (CHGB) is critical. Biochemical fractionation shows that native CHGB distributes nearly equally in soluble and membrane-bound forms, and both reconstitute highly selective anion channels in membrane. Confocal imaging resolves granular membrane components including proton pumps and CHGB in puncta on the cell surface after stimulated exocytosis. High pressure freezing immuno-EM reveals a major fraction of CHGB at granule membranes in rat pancreatic ß-cells. A cryo-EM structure of bCHGB dimer of a nominal 3.5 Å resolution delineates a central pore with end openings, physically sufficient for membrane-spanning and large single channel conductance. Together our data support that CHGB-containing (CHGB+) channels are characteristic of regulated secretion, and function in granule ion homeostasis near the plasma membrane or possibly in other intracellular processes. [ABSTRACT FROM AUTHOR]