Trial of the MIND Diet for Prevention of Cognitive Decline in Older Persons.

BACKGROUND: Findings from observational studies suggest that dietary patterns may offer protective benefits against cognitive decline, but data from clinical trials are limited. The Mediterranean-DASH Intervention for Neurodegenerative Delay, known as the MIND diet, is a hybrid of the Mediterranean diet and the DASH (Dietary Approaches to Stop Hypertension) diet, with modifications to include foods that have been putatively associated with a decreased risk of dementia. METHODS: We performed a two-site, randomized, controlled trial involving older adults without cognitive impairment but with a family history of dementia, a body-mass index (the weight in kilograms divided by the square of the height in meters) greater than 25, and a suboptimal diet, as determined by means of a 14-item questionnaire, to test the cognitive effects of the MIND diet with mild caloric restriction as compared with a control diet with mild caloric restriction. We assigned the participants in a 1:1 ratio to follow the intervention or the control diet for 3 years. All the participants received counseling regarding adherence to their assigned diet plus support to promote weight loss. The primary end point was the change from baseline in a global cognition score and four cognitive domain scores, all of which were derived from a 12-test battery. The raw scores from each test were converted to z scores, which were averaged across all tests to create the global cognition score and across component tests to create the four domain scores; higher scores indicate better cognitive performance. The secondary outcome was the change from baseline in magnetic resonance imaging (MRI)-derived measures of brain characteristics in a nonrandom sample of participants. RESULTS: A total of 1929 persons underwent screening, and 604 were enrolled; 301 were assigned to the MIND-diet group and 303 to the control-diet group. The trial was completed by 93.4% of the participants. From baseline to year 3, improvements in global cognition scores were observed in both groups, with increases of 0.205 standardized units in the MIND-diet group and 0.170 standardized units in the control-diet group (mean difference, 0.035 standardized units; 95% confidence interval, -0.022 to 0.092; P = 0.23). Changes in white-matter hyperintensities, hippocampal volumes, and total gray- and white-matter volumes on MRI were similar in the two groups. CONCLUSIONS: Among cognitively unimpaired participants with a family history of dementia, changes in cognition and brain MRI outcomes from baseline to year 3 did not differ significantly between those who followed the MIND diet and those who followed the control diet with mild caloric restriction. (Funded by the National Institute on Aging; ClinicalTrials.gov number, NCT02817074.).

Vimentin inhibits α-tubulin acetylation via enhancing α-TAT1 degradation to suppress the replication of human parainfluenza virus type 3.

We previously found that, among human parainfluenza virus type 3 (HPIV3) proteins, the interaction of nucleoprotein (N) and phosphoprotein (P) provides the minimal requirement for the formation of cytoplasmic inclusion bodies (IBs), which are sites of RNA synthesis, and that acetylated α-tubulin enhances IB fusion and viral replication. In this study, using immunoprecipitation and mass spectrometry assays, we determined that vimentin (VIM) specifically interacted with the N-P complex of HPIV3, and that the head domain of VIM was responsible for this interaction, contributing to the inhibition of IB fusion and viral replication. Furthermore, we found that VIM promoted the degradation of α-tubulin acetyltransferase 1 (α-TAT1), through its head region, thereby inhibiting the acetylation of α-tubulin, IB fusion, and viral replication. In addition, we identified a 20-amino-acid peptide derived from the head region of VIM that participated in the interaction with the N-P complex and inhibited viral replication. Our findings suggest that VIM inhibits the formation of HPIV3 IBs by downregulating α-tubulin acetylation via enhancing the degradation of α-TAT1. Our work sheds light on a new mechanism by which VIM suppresses HPIV3 replication.

Exploring correlations between microplastics, microorganisms, and water quality in an urban drinking water source.

The microplastic pollution in freshwater system is gradually becoming more severe, which has led to increasing attention on the distribution and potential harmful effects of microplastics. Moreover, microplastics may have an impact on river ecology and pose risks to ecosystems. Therefore, it is important to reveal this process. This study aimed to explore correlations between microplastics and free-living microorganisms in an urban drinking water source of Xiangjiang River by using multivariate statistical analysis. The results indicated that the abundance of microplastics (size 50 µm to 5 mm) in surface water and sediments ranged from 0.72 to 18.6 (mean ± SD: 7.32 ± 2.36) items L-1 and 26.3-302 (150 ± 75.6) items kg-1 dry weight (dw), respectively, suggesting potential microplastic pollution despite the protected status as a drinking water source. Higher microplastic abundances were observed in urban areas and the downstream of wastewater plants, with mostly granular shape, transparent and black color as well as 50-100 µm in size. The multivariate statistical analysis presented that the abundance of microplastics is not significantly correlated with water indicators, due to the complexity of the abundance data. The water indicators showed an obvious correlation with microplastics in colors of transparent and black, and smaller sizes of 50-100 µm. This is also true for microplastics and microorganisms in water and sediment. Proteobacteria was the main prokaryote in water and sediments, being positively correlated with 50-100 µm microplastics; while Chloroplastida was the dominated eukaryotes, presenting a weak correlation with smaller-size microplastics. Overall, when considering the properties of microplastics such as shape, color and size, the potential correlations with water indicators and microorganisms were more evident than abundance. This study provides new insights into the multivariate statistical analysis, explaining the potential correlations among microplastic properties, microorganisms and environmental factors in a river system.

Identification of sources and analysis of spatial distribution of soil heavy metals in northern China coal mining areas.

The mining and utilization of coal resources has not only promoted rapid economic development but also poses a potential threat to the ecological environment. The purpose of this study is to clarify the effects both of mining and land use types on the spatial distribution and particular sources of heavy metals in soil, using inverse distance weighted (IDW) and the Positive Matrix Factorization (PMF) model. A total of 99 topsoil and profile soil samples across different land use types and mining conditions were collected. The contamination of soil with Cd, Pb, and Hg in the research area was most severe, with the coefficient of variation (CV) of Hg being the largest, while also being heavily influenced by human activities. Severely polluted regions were mainly distributed in the center of the coal mining area, as well as near the highway. The contents of heavy metals for various land use patterns were ranked as follows: forestland > farmland > bare land > grassland > building land. Hg, Cd, Pb, Cr, and Zn had showed migration in the 0-60 cm depth range, and the enrichment factors (EFs) of Cd, Pb, Hg, and As in the soil profile were the most significant. The PMF demonstrated that the contributions of industrial activities and atmospheric deposition, transportation and mining activities, agricultural activities, and natural sources accounted for 31.25%, 28.13%, 22.24%, and 18.38%, respectively. The migration and deposition of atmospheric particulate matter from coal mining, transportation, and coal combustion under winds triggered heavy metal contamination in semi-arid areas of northern China. This phenomenon has important implications for the prevention and reduction of heavy metal pollution through various effective measures in coal-mining cities in northern China.

Impacts of DNA methylases and demethylases on the methylation and expression of Arabidopsis ethylene signal pathway genes.

Arabidopsis ethylene (ET) signal pathway plays important roles in various aspects. Cytosine DNA methylation is significant in controlling gene expression in plants. Here, we analyzed the bisulfite sequencing and mRNA sequencing data from Arabidopsis (de)methylase mutants met1, cmt3, drm1/2, ddm1, ros1-4, and rdd to investigate how DNA (de)methylases influence the DNA methylation and expression of Arabidopsis ET pathway genes. At least 32 genes are found to involved in Arabidopsis ET pathway by text mining. Among them, 14 genes are unmethylated or methylated with very low levels. ACS6 and ACS9 are conspicuously methylated within their upstream regions. The other 16 genes are predominantly methylated at the CG sites within gene body regions in wild-type plants, and mutation of MET1 resulted in almost entire elimination of the CG methylations. In addition, CG methylations within some genes are jointly maintained by MET1 and other (de)methylases. Analyses of mRNA-seq data indicated that some ET pathway genes were differentially expressed between wild-type and diverse mutants. PDF1.2, the marker gene of ET signal pathway, was found being regulated indirectly by the methylases. Eighty-two transposable elements (TEs) were identified to be associated to 15 ET pathway genes. ACS11 is found located in a heterochromatin region that contains 57 TEs, indicating its specific expression and regulation. Together, our results suggest that DNA (de)methylases are implicated in the regulation of CG methylation within gene body regions and transcriptional activity of some ET pathway genes and that maintenance of normal CG methylation is essential for ET pathway in Arabidopsis.

Clinical evaluation of metagenomic next-generation sequencing in unbiased pathogen diagnosis of urinary tract infection.

BACKGROUND: Early availability of pathogen identification in urinary tract infections (UTIs) has critical importance in disease management. Metagenomic next-generation sequencing (mNGS) has the potential to transform how acute and serious infections are diagnosed by offering unbiased and culture-free pathogen detection. However, clinical experience with application of the mNGS test is relatively limited. METHODS: We therefore established a MinION-based mNGS pathogens diagnostic platform and evaluated its potential for clinical implementation in UTIs with clinical samples. 213 urine samples from patients with suspected UTIs were included and subjected to mNGS testing using the MinION platform. mNGS results were compared to the gold standard of clinical culture and composite standard of combining clinical testing, confirmatory qPCR testing, and clinical adjudication by doctors. RESULTS: The mNGS exhibited a sensitivity of 81.4% and a specificity of 92.3%, along with a positive predictive value of 96.6%, a negative predictive value of 64.9%, and an overall accuracy of 84.4%, all of which were determined based on the gold standard of routine culture results. When assessed against the composite standard, the sensitivity and specificity both increased to 89.9% and 100%, respectively, while the accuracy rose to 92.4%. Notably, the positive predictive value and negative predictive value also saw improvements, reaching 100% and 76.8%, respectively. Moreover, this diagnostic platform successfully identified dsDNA viruses. Among the 65 culture-negative samples, the viral detection rate reached 33.8% (22/65) and was subsequently validated through qPCR. Furthermore, the automatic bioinformatics pipeline we developed enabled one-click analysis from data to results, leading to a significant reduction in diagnosis time. CONCLUSION: These results demonstrate that the pathogen detection performance of mNGS is sufficient for diagnostic testing in clinical settings. As the method is generally unbiased, it can improve diagnostic testing of UTIs and other microbial infections.

Multi-Temporal Hyperspectral Classification of Grassland Using Transformer Network.

In recent years, grassland monitoring has shifted from traditional field surveys to remote-sensing-based methods, but the desired level of accuracy has not yet been obtained. Multi-temporal hyperspectral data contain valuable information about species and growth season differences, making it a promising tool for grassland classification. Transformer networks can directly extract long-sequence features, which is superior to other commonly used analysis methods. This study aims to explore the transformer network's potential in the field of multi-temporal hyperspectral data by fine-tuning it and introducing it into high-powered grassland detection tasks. Subsequently, the multi-temporal hyperspectral classification of grassland samples using the transformer network (MHCgT) is proposed. To begin, a total of 16,800 multi-temporal hyperspectral data were collected from grassland samples at different growth stages over several years using a hyperspectral imager in the wavelength range of 400-1000 nm. Second, the MHCgT network was established, with a hierarchical architecture, which generates a multi-resolution representation that is beneficial for grass hyperspectral time series' classification. The MHCgT employs a multi-head self-attention mechanism to extract features, avoiding information loss. Finally, an ablation study of MHCgT and comparative experiments with state-of-the-art methods were conducted. The results showed that the proposed framework achieved a high accuracy rate of 98.51% in identifying grassland multi-temporal hyperspectral which outperformed CNN, LSTM-RNN, SVM, RF, and DT by 6.42-26.23%. Moreover, the average classification accuracy of each species was above 95%, and the August mature period was easier to identify than the June growth stage. Overall, the proposed MHCgT framework shows great potential for precisely identifying multi-temporal hyperspectral species and has significant applications in sustainable grassland management and species diversity assessment.

TMNet: A Two-Branch Multi-Scale Semantic Segmentation Network for Remote Sensing Images.

Pixel-level information of remote sensing images is of great value in many fields. CNN has a strong ability to extract image backbone features, but due to the localization of convolution operation, it is challenging to directly obtain global feature information and contextual semantic interaction, which makes it difficult for a pure CNN model to obtain higher precision results in semantic segmentation of remote sensing images. Inspired by the Swin Transformer with global feature coding capability, we design a two-branch multi-scale semantic segmentation network (TMNet) for remote sensing images. The network adopts the structure of a double encoder and a decoder. The Swin Transformer is used to increase the ability to extract global feature information. A multi-scale feature fusion module (MFM) is designed to merge shallow spatial features from images of different scales into deep features. In addition, the feature enhancement module (FEM) and channel enhancement module (CEM) are proposed and added to the dual encoder to enhance the feature extraction. Experiments were conducted on the WHDLD and Potsdam datasets to verify the excellent performance of TMNet.

Occurrence, sources, and risk assessment of pyrethroid insecticides in surface water and tap water from Taihu Lake, China.

Pyrethroid insecticides are one of the most widely used insecticides globally, posing a severe threat to human health and the environment. In this study, we applied high-throughput organic analysis testing combined with high-volume solid-phase extraction (Hi-throat/Hi-volume SPE) to elucidate the occurrence of 11 pyrethroid insecticides in lake water (n = 37), tributary river water (n = 15), and tap water (n = 6) in the Taihu Lake Basin. Permethrin was found to be the major contributing pyrethroid insecticide (detection rate = 100%). The concentrations of pyrethroid insecticides from different lake regions were revealed in the following descending order: southern > eastern > western > northern. The principal component analysis and multiple linear regression demonstrated that landscape maintenance, agricultural cultivation, and livestock breeding were the main sources of pyrethroid insecticides in the Taihu Lake surface water. Moreover, runoff input plays an important role in their accumulation, while the surrounding rivers contribute 2292 kg of pyrethroid insecticides to Taihu Lake annually. The risk assessment analysis demonstrated that pyrethroid insecticides pose a high risk to both the ecological environment and the surrounding human populations, thereby necessitating effective countermeasures. Furthermore, the pyrethroid insecticides in the Yangtze River Delta region have to be controlled. Overall, this is the first study focused on China that revealed the residue levels in water sources and tap water.

Assimilation behaviors and metabolite formations of estrone sulfate sodium (E1-3S) and 17ß-estradiol-3-O-sulfate sodium (E2-3S) in the wheat.

The conjugated steroid estrogens (CSEs), including estrone sulfate sodium (E1-3 S) and 17ß-estradiol-3-O-sulfate sodium (E2-3 S), exhibit distinct metabolic behaviors in the aqueous and soil environments. However, their assimilation behaviors and metabolite formations in plant bodies (shoots and roots) remain poorly understood. Therefore, this study used a modified plant hydroponic system to explore the efficiency with which wheat (Triticum acstivnm L.) assimilated the two estrogen conjugates, E1-3 S and E2-3 S. Results indicated the potential of wheat to absorb E1-3 S and E2-3 S, with their assimilation in the root being significantly higher (104-105 ng/g dw) than in the shoot (103-104 ng/g dw). E1-3 S de-sulfated and transformed to estrone (E1) at a rate of 4%-45% in the root's oxidative environment, whereas E2-3 S converted to E1-3 S at 210%-570%. However, the root-to-shoot transfer was impeded by a less potent metabolic activity within the shoot system. The co-exposure treatment revealed that E1 or 17ß-estradiol (E2) affects the assimilation of E1-3 S and E2-3 S by wheat, with E1 inhibiting E1-3 S assimilation and E2 promoting E2-3 S assimilation in wheat bodies. Nonetheless, free-form steroid estrogens (FSEs), which typically have a significant hormone action, can oxidative-damage the wheat tissues, producing a progressive wilting of wheat leaf and so limiting the transpiration process. Co-exposure initially increased the assimilation amounts of E1-3 S (particularly in shoots) and E2-3 S (in both roots and shoots), but these values rapidly declined as exposure duration increased. The combined effects of E1-3 S and E2-3 S exposure also increased their assimilation. These findings suggest the need for further investigation into the cumulative impact of environmental estrogen contaminants. The findings of present study can potentially guide the development of strategies to prevent and manage steroid estrogen contamination in agricultural contexts.

Reduced attention on restricted organochlorine pesticides, whereas still noteworthy of the impact on the deep soil and groundwater: a historical site study in southern China.

The use of hexachlorocyclohexanes (HCHs) in pesticides has been prohibited for decades in China. Since then, there have been urbanization and transformation of the functional areas of many sites, which were formerly involved in the HCH industry. However, it is possible that, unless properly managed, these sites may still contain HCH residues in the soil and thus pose a threat to the surrounding environment and the quality of groundwater. This study aimed to characterize soil residues in a typical site that was historically involved in HCH production in southern China, by analyzing the α-HCH, ß-HCH, and γ-HCH contents of the soil. The results suggested that HCHs persist in the environment and can have long-term effects. It was found that α-HCH and ß-HCH were present in many samples in concentrations that were comparable or higher than those specified by China's Class 1 screening values. The distribution of residues was significantly correlated with the historical HCH production activities in the areas. The characteristic ratios of α-HCH/γ-HCH and ß-HCH/(α + γ)-HCH at different soil depths were 1.4-3.7 and 0.21-1.04, respectively, which indicated the presence of significant localized residues of HCHs. The presence of HCHs in the soil suggested a downward migration, with concentrations rapidly decreasing in the upper layer soil (0-5 m), but a gradual increase in the deeper soil (5-14 m). HCHs were detected at depths exceeding 24 m, indicating heavy penetration. The proportions of γ-HCH and ß-HCH changed with increasing soil depth, which was related to their relatively volatile and stable molecular structures, respectively. The results strongly suggested that there is widespread contamination of both soil and groundwater by HCHs even after decades. The likelihood of residual HCHs in the soil should therefore be taken into full consideration during urban planning to limit risks to human and environmental health.

Development of high quality T1-weighted and diffusion tensor templates of the older adult brain in a common space.

High-quality T1-weighted (T1w) and diffusion tensor imaging (DTI) brain templates that are representative of the individuals under study enhance the accuracy of template-based neuroimaging investigations, and when they are also located in a common space they facilitate optimal integration of information on brain morphometry and diffusion characteristics. However, such multimodal templates have not been constructed for the brain of older adults. The purpose of this work was threefold: (A) to introduce an iterative method for construction of multimodal T1w and DTI templates that aims at maximizing the quality of each template separately as well as the spatial matching between templates, (B) to use this method to develop T1w and DTI templates of the older adult brain in a common space, and (C) to evaluate the performance of the method across iterations and compare it to the performance of state-of-the-art approaches based on multichannel registration. It was demonstrated that more iterations of the proposed method enhanced the characteristics and spatial matching of the resulting T1w and DTI templates. The templates of the older adult brain generated by the final iteration of the proposed method provided better delineation of brain structures, higher discriminability between tissues, and higher image sharpness near the cortex compared to templates generated with approaches employing multichannel registration. In addition, the spatial matching between the T1w and DTI templates constructed by the proposed method approximated the template alignment achieved with methods employing multichannel registration. Finally, when using the templates generated by the proposed method as references for spatial normalization of older adult T1w and DTI data, both the intra-modality inter-subject normalization precision and the inter-modality spatial matching were higher in most metrics than those achieved with templates constructed with other methods. Overall, the present work brought new insights into multimodal template construction, generated much-needed high quality T1w and DTI templates of the older adult brain in a common space, and conducted a thorough, quantitative evaluation of available multimodal template construction methods.

Mutual Regulation of Receptor-Like Kinase SIT1 and B'κ-PP2A Shapes the Early Response of Rice to Salt Stress.

The receptor-like kinase SIT1 acts as a sensor in rice (Oryza sativa) roots, relaying salt stress signals via elevated kinase activity to enhance salt sensitivity. Here, we demonstrate that Protein Phosphatase 2A (PP2A) regulatory subunit B'κ constrains SIT1 activity under salt stress. B'κ-PP2A deactivates SIT1 directly by dephosphorylating the kinase at Thr515/516, a salt-induced phosphorylation site in the activation loop that is essential for SIT1 activity. B'κ overexpression suppresses the salt sensitivity of rice plants expressing high levels of SIT1, thereby contributing to salt tolerance. B'κ functions in a SIT1 kinase-dependent manner. During early salt stress, activated SIT1 phosphorylates B'κ; this not only enhances its binding with SIT1, it also promotes B'κ protein accumulation via Ser502 phosphorylation. Consequently, by blocking SIT1 phosphorylation, B'κ inhibits and fine-tunes SIT1 activity to balance plant growth and stress adaptation.

Multifunctional Silica-Modified Hybrid Microgels Templated from Inverse Pickering Emulsions.

Microgels are regarded as soft colloids with environmental responsiveness. However, the majority of reported microgels are inherently hydrophilic, resulting in aqueous dispersions, and only used in water-based applications. Herein, we reported an efficient method for hybridization of poly(N-isopropylacrylamide) microgel by coating hydrophobic silica nanoparticles on their surface. The resultant hybrid microgel had switchable surface wettability and could be dispersed in both aqueous and oil phases. Meanwhile, the coated hydrophobic silica nanoparticles solved the difficulty in redispersing microgels caused by particle aggregation and film formation during the drying process, providing a significant advantage in dried storage. Furthermore, the introduction of hydrophobic silica nanoparticles endowed the hybrid microgel with a variety of applications, including cargo encapsulation, active release induced by emulsion reversion, and trace water absorption.

Screening of six cation exchange resins for high binding capacity, monomer purity and step yield: A case study.

Cation exchange (CEX) chromatography has been widely used as a polishing step in downstream processing of therapeutic monoclonal antibodies (mAbs). It has been well documented that the performance of a particular CEX resin heavily relies on buffer type, pH and conductivity. In this work, with a case study, we screened six commercial CEX resins under different pHs and conductivities. Initial screening was conducted on Tecan to find conditions under which high binding capacity was achieved. Next, performance of each resin was further evaluated using a small column under the identified optimum binding conditions. Variations in binding capacity and purity of eluate were observed among these resins. The adopted approach allows for identification of resins exhibiting good binding capacity, aggregate clearance and recovery.

A Bayesian hierarchical model for individual participant data meta-analysis of demand curves.

Individual participant data meta-analysis is a frequently used method to combine and contrast data from multiple independent studies. Bayesian hierarchical models are increasingly used to appropriately take into account potential heterogeneity between studies. In this paper, we propose a Bayesian hierarchical model for individual participant data generated from the Cigarette Purchase Task (CPT). Data from the CPT details how demand for cigarettes varies as a function of price, which is usually described as an exponential demand curve. As opposed to the conventional random-effects meta-analysis methods, Bayesian hierarchical models are able to estimate both the study-specific and population-level parameters simultaneously without relying on the normality assumptions. We applied the proposed model to a meta-analysis with baseline CPT data from six studies and compared the results from the proposed model and a two-step conventional random-effects meta-analysis approach. We conducted extensive simulation studies to investigate the performance of the proposed approach and discussed the benefits of using the Bayesian hierarchical model for individual participant data meta-analysis of demand curves.

Development and evaluation of a high performance T1-weighted brain template for use in studies on older adults.

Τhe accuracy of template-based neuroimaging investigations depends on the template's image quality and representativeness of the individuals under study. Yet a thorough, quantitative investigation of how available standardized and study-specific T1-weighted templates perform in studies on older adults has not been conducted. The purpose of this work was to construct a high-quality standardized T1-weighted template specifically designed for the older adult brain, and systematically compare the new template to several other standardized and study-specific templates in terms of image quality, performance in spatial normalization of older adult data and detection of small inter-group morphometric differences, and representativeness of the older adult brain. The new template was constructed with state-of-the-art spatial normalization of high-quality data from 222 older adults. It was shown that the new template (a) exhibited high image sharpness, (b) provided higher inter-subject spatial normalization accuracy and (c) allowed detection of smaller inter-group morphometric differences compared to other standardized templates, (d) had similar performance to that of study-specific templates constructed with the same methodology, and (e) was highly representative of the older adult brain.

EBV epitranscriptome reprogramming by METTL14 is critical for viral-associated tumorigenesis.

Epstein-Barr virus (EBV) is a ubiquitous oncogenic virus that induces many cancers. N6-Methyladenosine (m6A) modification regulates many cellular processes. We explored the role of m6A in EBV gene regulation and associated cancers. We have comprehensively defined m6A modification of EBV latent and lytic transcripts. Furthermore, m6A modification demonstrated a functional role in regulation of the stability of viral transcripts. The methyltransferase METTL14 was induced at the transcript and protein levels, and knock-down of METTL14 led to decreased expression of latent EBV transcripts. METTL14 was also significantly induced in EBV-positive tumors, promoted growth of EBV-transformed cells and tumors in Xenograft animal models. Mechanistically, the viral-encoded latent oncoprotein EBNA3C activated transcription of METTL14, and directly interacted with METTL14 to promote its stability. This demonstrated that EBV hijacks METTL14 to drive EBV-mediated tumorigenesis. METTL14 is now a new target for development of therapeutics for treatment of EBV-associated cancers.

EBNA3C facilitates RASSF1A downregulation through ubiquitin-mediated degradation and promoter hypermethylation to drive B-cell proliferation.

EBV latent antigen 3C (EBNA3C) is essential for EBV-induced primary B-cell transformation. Infection by EBV induces hypermethylation of a number of tumor suppressor genes, which contributes to the development of human cancers. The Ras association domain family isoform 1A (RASSF1A) is a cellular tumor suppressor, which regulates a broad range of cellular functions, including apoptosis, cell-cycle arrest, mitotic arrest, and migration. However, the expression of RASSF1A is lost in many human cancers by epigenetic silencing. In the present study, we showed that EBNA3C promoted B-cell transformation by specifically suppressing the expression of RASSF1A. EBNA3C directly interacted with RASSF1A and induced RASSF1A degradation via the ubiquitin-proteasome-dependent pathway. SCFSkp2, an E3-ubiquitin ligase, was recruited by EBNA3C to enhance RASSF1A degradation. Moreover, EBNA3C decreased the transcriptional activity of RASSF1A promoter by enhancing its methylation through EBNA3C-mediated modulation of DNMTs expression. EBNA3C also inhibited RASSF1A-mediated cell apoptosis, disrupted RASSF1A-mediated microtubule and chromosomal stability, and promoted cell proliferation by upregulating Cyclin D1 and Cyclin E expression. Our data provides new details, which sheds light on additional mechanisms by which EBNA3C can induce B-cell transformation. This will also facilitate the development of novel therapeutic approaches through targeting of the RASSF1A pathway.

Epigenetic upregulation of ssc-miR-124a following treatment with Clostridium perfringens beta2-toxin attenuates both apoptosis and inflammation in intestinal porcine epithelial cells.

Clostridium perfringens (C. perfringens) is a globally recognized zoonotic pathogen. It has been reported that the beta2-toxin produced by C. perfringens can cause a variety of gastrointestinal diseases and even systemic inflammation. MicroRNA-124a (miR-124a) has been reported to play important roles in the host response to pathogenic infection. Although C. perfringens beta2-toxin induced injury in intestinal porcine epithelial (IPEC-J2) cells has been established, the underlying molecular mechanism is not completely unraveled. Here we show that a significant upregulation of ssc-miR-124a in IPEC-J2 cells after beta2-toxin stimulation was associated with the MiR-124A-1 and MiR-124A-2 gene promoter demethylation status. Importantly, overexpression of ssc-miR-124a significantly increased cell proliferation and decreased apoptosis and cytotoxicity in beta2-toxin treated IPEC-J2 cells. Transfection of IPEC-J2 cells with ssc-miR-124a mimic suppressed beta2-toxin induced inflammation. On the contrary, ssc-miR-124a inhibitor promoted aggravation of cell apoptosis and excessive damage. Furthermore, rho-associated coiled-coil-containing protein kinase 1 (ROCK1) was identified as the direct target gene of ssc-miR-124a in IPEC-J2 cells and its siRNA transfection reversed the promotion of apoptosis and aggravation of cellular damage induced by ssc-miR-124a inhibitor. Overall, we speculated that the miR-124A-1/2 gene was epigenetically regulated in IPEC-J2 cells after beta2-toxin treatment. Upregulation of ssc-miR-124a may restrain ROCK1, and attenuate apoptosis and inflammation induced by beta2-toxin that prevent IPEC-J2 cells from severe damages. We discover a new molecular mechanism by which IPEC-J2 cells counteract beta2-toxin-induced damage through the ssc-miR-124a/ROCK1 axis partially.