SMGformer: integrating STL and multi-head self-attention in deep learning model for multi-step runoff forecasting.

Accurate runoff forecasting is of great significance for water resource allocation flood control and disaster reduction. However, due to the inherent strong randomness of runoff sequences, this task faces significant challenges. To address this challenge, this study proposes a new SMGformer runoff forecast model. The model integrates Seasonal and Trend decomposition using Loess (STL), Informer's Encoder layer, Bidirectional Gated Recurrent Unit (BiGRU), and Multi-head self-attention (MHSA). Firstly, in response to the nonlinear and non-stationary characteristics of the runoff sequence, the STL decomposition is used to extract the runoff sequence's trend, period, and residual terms, and a multi-feature set based on 'sequence-sequence' is constructed as the input of the model, providing a foundation for subsequent models to capture the evolution of runoff. The key features of the input set are then captured using the Informer's Encoder layer. Next, the BiGRU layer is used to learn the temporal information of these features. To further optimize the output of the BiGRU layer, the MHSA mechanism is introduced to emphasize the impact of important information. Finally, accurate runoff forecasting is achieved by transforming the output of the MHSA layer through the Fully connected layer. To verify the effectiveness of the proposed model, monthly runoff data from two hydrological stations in China are selected, and eight models are constructed to compare the performance of the proposed model. The results show that compared with the Informer model, the 1th step MAE of the SMGformer model decreases by 42.2% and 36.6%, respectively; RMSE decreases by 37.9% and 43.6% respectively; NSE increases from 0.936 to 0.975 and from 0.487 to 0.837, respectively. In addition, the KGE of the SMGformer model at the 3th step are 0.960 and 0.805, both of which can maintain above 0.8. Therefore, the model can accurately capture key information in the monthly runoff sequence and extend the effective forecast period of the model.

How online pro-environmental games affect users' pro-environmental behavioural intentions? - Insights from Ant Forest.

Information technology has created new ways for people to participate in environmental protection. Ant Forest, a pro-environmental game that actively encourages users to take part in offline environmental activities through online gaming content, has gained great popularity. However, research in this area is still limited, and the effect of playing such games on users' pro-environmental intentions in real life and the underlying mechanisms are unknown. Therefore, this research draws from Persuasive Systems Design model and value theory and examines the relationships between design features of Ant Forest, users' values and their pro-environmental behavioural intentions. The research model is empirically tested by survey data from 428 Ant Forest users. Results indicate that users' self-enhancement values, including utilitarian, hedonic and social values, and self-transcendence values, including altruistic and biospheric values, all positively affect their continued gameplay intentions and therefore affect their pro-environmental behavioural intentions. Additionally, the primary task support, social support, dialogue support, and system credibility of Ant Forest have varying positive effects on users' perceived values.

Stiffness promotes cell migration, invasion, and invadopodia in nasopharyngeal carcinoma by regulating the WT-CTTN level.

Matrix stiffness potently promotes the malignant phenotype in various biological contexts. Therefore, identification of gene expression to participate in mechanical force signals transduced into downstream biochemical signaling will contribute substantially to the advances in nasopharyngeal carcinoma (NPC) treatment. In the present study, we detected that cortactin (CTTN) played an indispensable role in matrix stiffness-induced cell migration, invasion, and invadopodia formation. Advances in cancer research have highlighted that dysregulated alternative splicing contributes to cancer progression as an oncogenic driver. However, whether WT-CTTN or splice variants (SV1-CTTN or SV2-CTTN) regulate matrix stiffness-induced malignant phenotype is largely unknown. We proved that alteration of WT-CTTN expression modulated matrix stiffness-induced cell migration, invasion, and invadopodia formation. Considering that splicing factors might drive cancer progression through positive feedback loops, we analyzed and showed how the splicing factor PTBP2 and TIA1 modulated the production of WT-CTTN. Moreover, we determined that high stiffness activated PTBP2 expression. Taken together, our findings showed that the PTBP2-WT-CTTN level increases upon stiffening and then promotes cell migration, invasion, and invadopodia formation in NPC.

The Transcription Factor Ets1 Influences Axonal Growth via Regulation of Lcn2.

Transcription factors are essential for the development and regeneration of the nervous system. The current study investigated key regulatory transcription factors in rat spinal cord development via RNA sequencing. The hub gene Ets1 was highly expressed in the spinal cord during the embryonic period, and then its expression decreased during spinal cord development. Knockdown of Ets1 significantly increased the axonal growth of cultured spinal cord neurons. Luciferase reporter assays and chromatin immunoprecipitation assays indicated that Ets1 could directly bind to the Lcn2 promoter and positively regulate Lcn2 transcription. In conclusion, these findings provide the first direct evidence that Ets1 regulates axon growth by controlling Lcn2 expression, and Ets1 may be a novel therapeutic target for axon regeneration in the central nervous system.

Factor diagnosis and governance strategies of ship oil spill accidents based on formal concept analysis.

Ship oil spill accidents have a prolonged duration, complex consequences, challenging cleaning and repairing efforts, and pose a significant threat to the environment, economy, and society. Eliminating irrelevant information and identifying key factors using traditional methods is challenging due to the complexity of the causes of ship oil spill accidents. To address this, this article sorts out the accident databases of the International Tanker Owners Pollution Federation (ITOPF) and eight national maritime administration agencies, and innovatively constructs a formal concept analysis (FCA) model based on reports of 100-plus ship oil spill accidents. The model results prove that improper operation, less complete ship equipment, large tonnage, and poor navigation conditions are the key factors. The different causal rules of oil spills in collision/contact, grounding, fire/explosion, and foundering are further compared and analyzed. Finally, corresponding improvement measures are put forward for the key factors of oil spills and different causal rules.

WSCNet: Biomedical Image Recognition for Cell Encapsulated Microfluidic Droplets.

Microfluidic droplets accommodating a single cell as independent microreactors are frequently demanded for single-cell analysis of phenotype and genotype. However, challenges exist in identifying and reducing the covalence probability (following Poisson's distribution) of more than two cells encapsulated in one droplet. It is of great significance to monitor and control the quantity of encapsulated content inside each droplet. We demonstrated a microfluidic system embedded with a weakly supervised cell counting network (WSCNet) to generate microfluidic droplets, evaluate their quality, and further recognize the locations of encapsulated cells. Here, we systematically verified our approach using encapsulated droplets from three different microfluidic structures. Quantitative experimental results showed that our approach can not only distinguish droplet encapsulations (F1 score > 0.88) but also locate each cell without any supervised location information (accuracy > 89%). The probability of a "single cell in one droplet" encapsulation is systematically verified under different parameters, which shows good agreement with the distribution of the passive method (Residual Sum of Squares, RSS < 0.5). This study offers a comprehensive platform for the quantitative assessment of encapsulated microfluidic droplets.

SCARB1 in extracellular vesicles promotes NPC metastasis by co-regulating M1 and M2 macrophage function.

Distant metastasis is currently the main factor affecting the prognosis of nasopharyngeal carcinoma (NPC), and understanding the mechanisms of metastasis and identifying reliable therapeutic targets are critical for improving prognosis and achieving clinical translation. Macrophages, as important immune cells in the tumor microenvironment (TME), have been shown to regulate metastasis. And extracellular vesicles (EVs) secreted by stromal cells and tumor cells play the important role in intercellular communication in the tumor microenvironment. However, the role of NPC-EVs on macrophages and their function in regulating macrophages to affect metastasis has not been fully clarified. In this study, we report that NPC-EVs can be uptake by macrophages and alter macrophage polarization, for the first time, we identified the genes implicated in these regulatory functions: SCARB1, HAAO, and CYP1B1. Moreover, we found that SCARB1 was positively associated with metastasis and poor prognosis of NPC. Interestingly, we found that SCARB1-rich EVs promoted M1 macrophages ferroptosis to decrease M1 macrophages infiltration by upregulating the HAAO level while decreasing phagocytosis of M2 macrophages by upregulating the CYP1B1 level. Finally, we identified the SCARB1-binding gene KLF9, which is involved in the transcription of HAAO and CYP1B1. Our findings showed that SCARB1-EVs promoted metastasis by co-regulating M1 and M2 macrophage function. The related mechanism will provide a new therapeutic strategy to help patients with NPC improve their prognosis.

RCN2 promotes Nasopharyngeal carcinoma progression by curbing Calcium flow and Mitochondrial apoptosis.

OBJECTIVE: Evidence suggests that calcium release from the endoplasmic reticulum (ER) can be induced to cause calcium overload, which in turn can trigger mitochondrial-dependent apoptosis. Dysregulation of systemic calcium homeostasis and changing levels of calcium-binding proteins have been shown to be associated with the malignant behavior of tumors. However, the precise molecular mechanism underlying Nasopharyngeal carcinoma (NPC) remains uncertain. METHODS: Reticulocalbin (RCN2) expression in NPC was assessed using GEO database, western blot analysis and qRT-PCR. Apoptosis was assessed using flow cytometric analysis and the expression levels of apoptosis-related proteins were determined using western blot analysis. Intracellular calcium ion concentrations were measured using fluorescence imaging. The findings from these analyses were validated in vitro using nude mice models. Luciferase and ChIP assays were used to measure transcriptional regulation. Clinical significance was evaluated using tissue microarray analysis (n=150). RESULTS: Our results showed that RCN2 promotes malignancy by causing Ca2+ flow imbalance, which leads to the initiation of the stress-mediated mitochondrial apoptosis pathway. We demonstrate that calreticulin (CALR) resides primarily in the endoplasmic reticulum and interacts with RCN2. Moreover, the transcription factors YY1 and homeobox protein goosecoid (GSC) both contribute to the initiation of RCN2 transcription by directly binding to the predicted promoter region of RCN2. Finally, high expression of RCN2 combined with high expression of GSC and YY1 may serve as an important clinical biomarker of poor prognosis in patients with NPC. CONCLUSION: YY1 and GSC are upstream regulators of RCN2, involved in mitochondrial calcium overload and stress-induced mitochondrial apoptosis. Thus, they can play significant role in the malignant development of NPCs.

RNA Modifications Meet Tumors.

RNA modifications occur through the whole process of gene expression regulation, including transcription, translation, and post-translational processes. They are closely associated with gene expression, RNA stability, and cell cycle. RNA modifications in tumor cells play a vital role in tumor development and metastasis, changes in the tumor microenvironment, drug resistance in tumors, construction of tumor cell-cell "internet", etc. Several types of RNA modifications have been identified to date and have various effects on the biological characteristics of different tumors. In this review, we discussed the function of RNA modifications, including N 6-methyladenine (m6A), 5-methylcytosine (m5C), N 7-methyladenosine (m7G), N 1-methyladenosine (m1A), pseudouridine (Ψ), and adenosine-to-inosine (A-to-I), in the microenvironment and therapy of solid and liquid tumors.

A prospective study on the difference of clinical outcomes between elderly and adult patients with allergic rhinitis.

OBJECTIVE: The guiding significance of existing guidelines for the diagnosis, treatment and health management of AR in elderly patients is unclear. The aim of this study is to analyze the clinical characteristics and therapeutic effects of elderly and adult AR patients by prospective study. METHODS: A total of 131 AR patients were recruited and divided into elderly group and adult group according to age. After receiving the same pharmacological treatment for 4 weeks, the differences of the two groups in clinical scores including TNSS-4, RQLQ and VAS were compared. RESULTS: After 4 weeks treatment, all clinical scores in the adult group were improved compared with the baseline levels, while in the elderly group, only the TNSS-4 score was significantly reduced, and the RQLQ and VAS scores were not significantly improved. The changes of TNSS-4, RQLQ, and VAS scores in the elderly group were significantly inferior to those in the adult group (LS mean differences were 1.60, 8.80, and 11.10, respectively; P < 0.001). CONCLUSION: We confirmed that elderly and adult AR patients had different clinical characteristics and outcomes, and the degree of improvement in the adult group was significantly better than that in the elderly group. Therefore, it is urgent for us to establish a clinical guideline suitable for the elderly AR population to give more scientific and reasonable recommendations for diagnosis and treatment.

Extracellular vesicles rich in HAX1 promote angiogenesis by modulating ITGB6 translation.

Tumour-associated angiogenesis plays a critical role in metastasis, the main cause of malignancy-related death. Extracellular vesicles (EVs) can regulate angiogenesis to participate in tumour metastasis. Our previous study showed that EVs rich in HAX1 are associated with in metastasis of nasopharyngeal carcinoma (NPC). However, the mechanism by which HAX1 of EVs promotes metastasis and angiogenesis is unclear. In this study, we demonstrated that EVs rich in HAX1 promote angiogenesis phenotype by activating the FAK pathway in endothelial cells (ECs) by increasing expression level of ITGB6. The expression level of HAX1 is markedly correlated with microvessel density (MVDs) in NPC and head and neck cancers based on an analysis of IHC. In addition to a series of in vitro cellular analyses, in vivo models revealed that HAX1 was correlated with migration and blood vessel formation of ECs, and metastasis of NPC. Using ribosome profiling, we found that HAX1 regulates the FAK pathway to influence microvessel formation and promote NPC metastasis by enhancing the translation efficiency of ITGB6. Our findings demonstrate that HAX1 can be used as an important biomarker for NPC metastasis, providing a novel basis for antiangiogenesis therapy in clinical settings.

Evolution of the ErbB gene family and analysis of regulators of Egfr expression during development of the rat spinal cord.

Egfr, a member of the ErbB gene family, plays a critical role in tissue development and homeostasis, wound healing, and disease. However, expression and regulators of Egfr during spinal cord development remain poorly understood. In this study, we investigated ErbB evolution and analyzed co-expression modules, miRNAs, and transcription factors that may regulate Egfr expression in rats. We found that ErbB family members formed via Egfr duplication in the ancient vertebrates but diverged after speciation of gnathostomes. We identified a module that was co-expressed with Egfr, which involved cell proliferation and blood vessel development. We predicted 25 miRNAs and nine transcription factors that may regulate Egfr expression. Dual-luciferase reporter assays showed six out of nine transcription factors significantly affected Egfr promoter reporter activity. Two of these transcription factors (KLF1 and STAT3) inhibited the Egfr promoter reporter, whereas four transcription factors (including FOXA2) activated the Egfr promoter reporter. Real-time PCR and immunofluorescence experiments showed high expression of FOXA2 during the embryonic period and FOXA2 was expressed in the floor plate of the spinal cord, suggesting the importance of FOXA2 during embryonic spinal cord development. Considering the importance of Egfr in embryonic spinal cord development, wound healing, and disease (specifically in cancer), regulatory elements identified in this study may provide candidate targets for nerve regeneration and disease treatment in the future.

Let-7i-5p promotes a malignant phenotype in nasopharyngeal carcinoma via inhibiting tumor-suppressive autophagy.

MicroRNAs (miRNAs) regulate gene expression to participate in carcinogenesis and tumor progression. Therefore, identification of a malignant phenotype associated with miRNAs and therapeutic targets will contribute substantially in improving nasopharyngeal carcinoma (NPC) treatment. In this study, we demonstrated that overexpression of let-7i-5p promotes the malignant phenotype by acting as an autophagy suppressor by targeting ATG10 and ATG16L1 in NPC. Expression levels of let-7i-5p were markedly increased in NPC and head and neck cancers based on an analysis of the Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) databases. Using a cohort comprising 150 NPC tissues, we found that let-7i-5p was correlated with advanced stage, recurrence, metastasis, lymph node metastasis, and poor clinical outcomes. In addition to a series of in vitro cellular analyses, in vivo mouse tumor models revealed that let-7i-5p inhibits autophagy and promotes the malignant phenotype of NPC by targeting ATG10 and ATG16L1. Our findings demonstrate that let-7i-5p may represent a promising therapeutic target for NPC treatment.

AMPK-mTOR-Mediated Activation of Autophagy Promotes Formation of Dormant Polyploid Giant Cancer Cells.

Dormant cancer cells that survive anticancer therapy can lead to cancer recurrence and disseminated metastases that prove fatal in most cases. Recently, specific dormant polyploid giant cancer cells (PGCC) have drawn our attention because of their association with the clinical risk of nasopharyngeal carcinoma (NPC) recurrence, as demonstrated by previous clinical data. In this study, we report the biological properties of PGCC, including mitochondrial alterations, and reveal that autophagy is a critical mechanism of PGCC induction. Moreover, pharmacologic or genetic inhibition of autophagy greatly impaired PGCC formation, significantly suppressing metastasis and improving survival in a mouse model. Mechanistically, chemotherapeutic drugs partly damaged mitochondria, which then produced low ATP levels and activated autophagy via the AMPK-mTOR pathway to promote PGCC formation. Analysis of the transcriptional and epigenetic landscape of PGCC revealed overexpression of RIPK1, and the scaffolding function of RIPK1 was required for AMPK-mTOR pathway-induced PGCC survival. High numbers of PGCCs correlated with shorter recurrence time and worse survival outcomes in patients with NPC. Collectively, these findings suggest a therapeutic approach of targeting dormant PGCCs in cancer. SIGNIFICANCE: Pretreatment with an autophagy inhibitor before chemotherapy could prevent formation of therapy-induced dormant polyploid giant cancer cells, thereby reducing recurrence and metastasis of nasopharyngeal carcinoma.

A rapid lateral flow immunoassay strip for detection of SARS-CoV-2 antigen using latex microspheres.

BACKGROUND: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a highly infectious and concealed virus that causes pneumonia, severe acute respiratory syndrome, and even death. Although the epidemic has been controlled since the development of vaccines and quarantine measures, many people are still infected, particularly in third-world countries. Several methods have been developed for detection of SARS-CoV-2, but owing to its price and efficiency, the immune strip could be a better method for the third-world countries. METHODS: In this study, two antibodies were linked to latex microspheres, using 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide, as the bridge to decrease the cost further and improve the detection performance. The specificity of the lateral flow immunoassay strip (LFIA) was tested by several common viruses and respiratory bacterial infections. Besides, the reproducibility and stability of the LFIAs were tested on the same batch of test strips. Under optimal conditions, the sensitivity of LFIA was determined by testing different dilutions of the positive specimens. RESULTS: The proposed LFIAs were highly specific, and the limit of detection was as low as 25 ng/mL for SARS-CoV-2 antigens. The clinical applicability was evaluated with 659 samples (230 positive and 429 negative samples) by using both LFIA and rRT-PCR. Youden's index (J) was used to assess the performance of these diagnostic tests. The sensitivity and specificity were 98.22% and 97.93%, respectively, and J is 0.9615. The sensitivity and specificity were 98.22% and 97.93%, respectively, and J is 0.9615. In addition, the consistency of our proposed LFIA was analyzed using Cohen's kappa coefficient (κ = 0.9620). CONCLUSION: We found disease stage, age, gender, and clinical manifestations have only a slight influence on the diagnosis. Therefore, the lateral flow immunoassay SARS-CoV-2 antigen test strip is suitable for point-of-care detection and provides a great application for SARS-CoV-2 epidemic control in the third-world countries.

Local Integral Regression Network for Cell Nuclei Detection.

Nuclei detection is a fundamental task in the field of histopathology image analysis and remains challenging due to cellular heterogeneity. Recent studies explore convolutional neural networks to either isolate them with sophisticated boundaries (segmentation-based methods) or locate the centroids of the nuclei (counting-based approaches). Although these two methods have demonstrated superior success, their fully supervised training demands considerable and laborious pixel-wise annotations manually labeled by pathology experts. To alleviate such tedious effort and reduce the annotation cost, we propose a novel local integral regression network (LIRNet) that allows both fully and weakly supervised learning (FSL/WSL) frameworks for nuclei detection. Furthermore, the LIRNet can output an exquisite density map of nuclei, in which the localization of each nucleus is barely affected by the post-processing algorithms. The quantitative experimental results demonstrate that the FSL version of the LIRNet achieves a state-of-the-art performance compared to other counterparts. In addition, the WSL version has exhibited a competitive detection performance and an effortless data annotation that requires only 17.5% of the annotation effort.

ER resident protein 44 promotes malignant phenotype in nasopharyngeal carcinoma through the interaction with ATP citrate lyase.

BACKGROUND: Nasopharyngeal carcinoma (NPC) is one of the most common malignancy in head and neck. With the development of treatments, the prognosis has improved these years, but metastasis is still the main cause of treatment failure. The endoplasmic reticulum (ER) resident protein 44 is a UPR-induced ER protein of the protein disulphide isomerase (PDI) family. This study investigated the role of ERp44 in NPC progression. METHODS: Firstly, immunohistochemistry, western blot and qRT-PCR were used to investigate the expression of ERp44 in NPC samples and cell lines. We analyzed 44 NPC samples for ERp44 expression and investigated the association between its expression level with clinicopathologic parameters. Then we took CCK8, Transwell migration assay and used the zebrafish model to access the role of ERp44 on the malignant phenotype in NPC cells. Secondly, we used co-IP to gain the proteins that interact with ERp44 and took proteomic analysis. Furthermore, we successfully constructed the mutant variants of ERp44 and found the interaction domain with ATP citrate lyase(ACLY). Lastly, we subcutaneously injected NPC cells into nude mice and took immunohistochemistry to exam the expression of ACLY and ERp44. Then we used western blot to detect the expression level of epithelial-mesenchymal transition (EMT) markers. RESULTS: In the present study, we found ERp44 was elevated in NPC tissues and correlated with clinical stages and survive state of the patients. In vitro, the downregulation of ERp44 in NPC cells (CNE2, 5-8F) could suppress cells proliferation and migration. After that, we recognized that ACLY might be a potential target that could interact with ERp44. We further constructed the mutant variants of ERp44 and found the interaction domain with ACLY. The promotion of ERp44 on cell migration could be inhibited when ACLY was knocked down. More importantly, we also observed that the interaction of ERp44 with ACLY, especially the thioredoxin region in ERp44 play a vital role in regulating EMT. Lastly, we found ERp44 was positively correlated with the expression of ACLY and could promote NPC cells growth in nude mice. CONCLUSION: Our data indicated that ERp44 participates in promoting NPC progression through the interaction with ACLY and regulation of EMT.

Biocompatibility and biosafety of butterfly wings for the clinical use of tissue-engineered nerve grafts.

In a previous study, we used natural butterfly wings as a cell growth matrix for tissue engineering materials and found that the surface of different butterfly wings had different ultramicrostructures, which can affect the qualitative growth of cells and regulate cell growth, metabolism, and gene expression. However, the biocompatibility and biosafety of butterfly wings must be studied. In this study, we found that Sprague-Dawley rat dorsal root ganglion neurons could grow along the structural stripes of butterfly wings, and Schwann cells could normally attach to and proliferate on different species of butterfly wings. The biocompatibility and biosafety of butterfly wings were further examined through subcutaneous implantation in Sprague-Dawley rats, intraperitoneal injection in Institute of Cancer Research mice, intradermal injection in rabbits, and external application to guinea pigs. Our results showed that butterfly wings did not induce toxicity, and all examined animals exhibited normal behaviors and no symptoms, such as erythema or edema. These findings suggested that butterfly wings possess excellent biocompatibility and biosafety and can be used as a type of tissue engineering material. This study was approved by the Experimental Animal Ethics Committee of Jiangsu Province of China (approval No. 20190303-18) on March 3, 2019.

MIR106A-5p upregulation suppresses autophagy and accelerates malignant phenotype in nasopharyngeal carcinoma.

Dysregulated microRNAs (miRNAs) are involved in carcinoma progression, metastasis, and poor prognosis. We demonstrated that in nasopharyngeal carcinoma (NPC), transactivated MIR106A-5p promotes a malignant phenotype by functioning as a macroautophagy/autophagy suppressor by targeting BTG3 (BTG anti-proliferation factor 3) and activating autophagy-regulating MAPK signaling. MIR106A-5p expression was markedly increased in NPC cases based on quantitative real-time PCR, miRNA microarray, and TCGA database analysis findings. Moreover, MIR106A-5p was correlated with advanced stage, recurrence, and poor clinical outcomes in NPC patients. In addition to three-dimensional cell culture assays, zebrafish and BALB/c mouse tumor models revealed that overexpressed MIR106A-5p targeted BTG3 and accelerated the NPC malignant phenotype by inhibiting autophagy. BTG3 promoted autophagy, and its expression was correlated with poor prognosis in NPC. Attenuation of autophagy, mediated by the MIR106A-5p-BTG3 axis, occurred because of MAPK pathway activation. MIR106A-5p overexpression in NPC was due to increased transactivation by EGR1 and SOX9. Our findings may lead to novel insights into the pathogenesis of NPC.Abbreviations: ACTB: actin beta; ATG: autophagy-related; ATG5: autophagy related 5; BLI: bioluminescence; BTG3: BTG anti-proliferation factor 3; CASP3: caspase 3; ChIP: chromatin immunoprecipitation; CQ: chloroquine; Ct: threshold cycle; DAPI: 4',6-diamidino-2-phenylindole; DiL: 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate; EBSS: Earle's balanced salt solution; EGR1: early growth response 1; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GEO: Gene Expression Omnibus; GFP: green fluorescent protein; IF: immunofluorescence; IHC: immunohistochemistry; ISH: in situ hybridization; MAP1LC3B: microtubule associated protein 1 light chain 3 beta; MIR106A-5p: microRNA 106a-5p; miRNAs: microRNAs; MKI67: marker of proliferation ki-67; mRNA: messenger RNA; MTOR: mechanistic target of rapamycin kinase; NPC: nasopharyngeal carcinoma; qRT-PCR: quantitative real-time PCR; siRNA: small interfering RNA; SOX9: SRY-box transcription factor 9; SQSTM1: sequestosome 1; TCGA: The Cancer Genome Atlas; WB: western blot.

Copper Redox Cycling Inhibits Aß Fibre Formation and Promotes Fibre Fragmentation, while Generating a Dityrosine Aß Dimer.

Oxidative stress and the formation of plaques which contain amyloid-ß (Aß) peptides are two key hallmarks of Alzheimer's disease (AD). Dityrosine is found in the plaques of AD patients and Aß dimers have been linked to neurotoxicity. Here we investigate the formation of Aß dityrosine dimers promoted by Cu2+/+ Fenton reactions. Using fluorescence measurements and UV absorbance, we show that dityrosine can be formed aerobically when Aß is incubated with Cu2+ and hydrogen-peroxide, or in a Cu2+ and ascorbate redox mixture. The dityrosine cross-linking can occur for both monomeric and fibrillar forms of Aß. We show that oxidative modification of Aß impedes the ability for Aß monomer to form fibres, as indicated by the amyloid specific dye Thioflavin T (ThT). Transmission electron microscopy (TEM) indicates the limited amyloid assemblies that form have a marked reduction in fibre length for Aß(1-40). Importantly, the addition of Cu2+ and a reductant to preformed Aß(1-40) fibers causes their widespread fragmentation, reducing median fibre lengths from 800 nm to 150 nm upon oxidation. The processes of covalent cross-linking of Aß fibres, dimer formation, and fibre fragmentation within plaques are likely to have a significant impact on Aß clearance and neurotoxicity.