Accelerating protein aggregation and amyloid fibrillation for rapid inhibitor screening.

The accumulation and deposition of amyloid fibrils, also known as amyloidosis, in tissues and organs of patients has been found to be linked to numerous devastating neurodegenerative diseases. The aggregation of proteins to form amyloid fibrils, however, is a slow pathogenic process, and is a major issue for the evaluation of the effectiveness of inhibitors in new drug discovery and screening. Here, we used microdroplet reaction technology to accelerate the amyloid fibrillation process, monitored the process to shed light on the fundamental mechanism of amyloid self-assembly, and demonstrated the value of the technology in the rapid screening of potential inhibitor drugs. Proteins in microdroplets accelerated to form fibrils in milliseconds, enabling an entire cycle of inhibitor screening for Aß40 within 3 minutes. The technology would be of broad interest to drug discovery and therapeutic design to develop treatments for diseases associated with protein aggregation and fibrillation.

Erasable and Field Programmable DNA Circuits Based on Configurable Logic Blocks.

DNA is commonly employed as a substrate for the building of artificial logic networks due to its excellent biocompatibility and programmability. Till now, DNA logic circuits are rapidly evolving to accomplish advanced operations. Nonetheless, nowadays, most DNA circuits remain to be disposable and lack of field programmability and thereby limits their practicability. Herein, inspired by the Configurable Logic Block (CLB), the CLB-based erasable field-programmable DNA circuit that uses clip strands as its operation-controlling signals is presented. It enables users to realize diverse functions with limited hardware. CLB-based basic logic gates (OR and AND) are first constructed and demonstrated their erasability and field programmability. Furthermore, by adding the appropriate operation-controlling strands, multiple rounds of programming are achieved among five different logic operations on a two-layer circuit. Subsequently, a circuit is successfully built to implement two fundamental binary calculators: half-adder and half-subtractor, proving that the design can imitate silicon-based binary circuits. Finally, a comprehensive CLB-based circuit is built that enables multiple rounds of switch among seven different logic operations including half-adding and half-subtracting. Overall, the CLB-based erasable field-programmable circuit immensely enhances their practicability. It is believed that design can be widely used in DNA logic networks due to its efficiency and convenience.

Insights into the response of electroactive biofilm with petroleum hydrocarbons degradation ability to quorum sensing signals.

Bioelectrochemical technologies based on electroactive biofilms (EAB) are promising for petroleum hydrocarbons (PHs) remediation as anode can serve as inexhaustible electron acceptor. However, the toxicity of PHs might inhibit the formation and function of EABs. Quorum sensing (QS) is ideal for boosting the performance of EABs, but its potential effects on reshaping microbial composition of EABs in treating PHs are poorly understood. Herein, two AHL signals, C4-HSL and C12-HSL, were employed to promote EABs for PHs degradation. The start-times of AHL-mediated EABs decreased by 18-26%, and maximum current densities increased by 28-63%. Meanwhile, the removal of total PHs increased to over 90%. AHLs facilitate thicker and more compact biofilm as well as higher viability. AHLs enhanced the electroactivity and direct electron transfer capability. The total abundance of PH-degrading bacteria increased from 52.05% to 75.33% and 72.02%, and the proportion of electroactive bacteria increased from 26.14% to 62.72% and 63.30% for MFC-C4 and MFC-C12. Microbial networks became more complex, aggregated, and stable with addition of AHLs. Furthermore, AHL-stimulated EABs showed higher abundance of genes related to PHs degradation. This work advanced our understanding of AHL-mediated QS in maintaining the stable function of microbial communities in the biodegradation process of petroleum hydrocarbons.

Cellulose Nanofibrils of High Immunoaffinity for Efficient Enrichment of Small Extracellular Vesicles.

Extracellular vesicles (EVs), crucial in facilitating the transport of diverse molecular cargoes for intercellular communication, have shown great potential in diagnostics, therapeutics, and drug delivery. The challenge of developing effective preparation methods for EVs is heightened by their intrinsic heterogeneity and complexity. Here, a novel strategy for high EV enrichment is developed by utilizing EV-affinitive-modified cellulose nanofibrils. Specifically, modified cellulose with rich carboxyl groups has outstanding dispersing properties, able to be dispersed into cellulose nanofibrils in solution. These cellulose nanofibrils are utilized as scaffolds for the immobilization of EV-affinitive antibody of CD63 by chemical conjugation. The CD63-modified nanofibrils demonstrate a superior EV capture efficiency of 86.4% compared with other reported methods. The high performance of this system is further validated by the efficient capture of EVs from biological blood plasma, allowing the detection of bioactive markers from EV-derived miRNAs and proteins. The authors envision that these modified cellulose nanofibrils of enhanced capability on EV enrichment will open new avenues in various biomedical applications.

Electrospray-assisted cryo-EM sample preparation to mitigate interfacial effects.

Addressing interfacial effects during specimen preparation in cryogenic electron microscopy remains challenging. Here we introduce ESI-cryoPrep, a specimen preparation method based on electrospray ionization in native mass spectrometry, designed to alleviate issues associated with protein denaturation or preferred orientation induced by macromolecule adsorption at interfaces. Through fine-tuning spraying parameters, we optimized protein integrity preservation and achieved the desired ice thickness for analyzing target macromolecules. With ESI-cryoPrep, we prepared high-quality cryo-specimens of five proteins and obtained three-dimensional reconstructions at near-atomic resolution. Our findings demonstrate that ESI-cryoPrep effectively confines macromolecules within the middle of the thin layer of amorphous ice, facilitating the preparation of blotting-free vitreous samples. The protective mechanism, characterized by the uneven distribution of charged biomolecules of varying sizes within charged droplets, prevents the adsorption of target biomolecules at air-water or graphene-water interfaces, thereby avoiding structural damage to the protein particles or the introduction of dominant orientation issues.

Arenethiolate-Catalyzed 1,5-HAT of Aryl Halides: Synthesis of γ-Spirolactams.

γ-Spirolactam is a privileged building block that is found in a wide range of natural products and bioactive compounds. Herein, we report an arenethiolate-catalyzed 1,5-HAT of aryl halides to obtain γ-spirolactams through a SET reduction/intramolecular 1,5-HAT/cyclization/HAT process. This protocol features metal-free conditions and a broad substrate scope, furnishing the γ-spirolactams in moderate to excellent yields. Notably, aryl bromides, aryl chlorides and even aryl fluorides are well tolerated in this transformation. A mechanism involving arenethiolate as a SET catalyst is proposed based on DFT calculation.

Synthesis and Characterization of Superhydrophobic Epoxy Resin Coating with SiO2@CuO/HDTMS for Enhanced Self-Cleaning, Photocatalytic, and Corrosion-Resistant Properties.

The exceptional corrosion resistance and combined physical and chemical self-cleaning capabilities of superhydrophobic photocatalytic coatings have sparked significant interest among researchers. In this paper, we propose an economical and eco-friendly superhydrophobic epoxy resin coating that incorporates SiO2@CuO/HDTMS nanoparticles modified with Hexadecyltrimethoxysilane (HDTMS). The application of superhydrophobic coatings effectively reduces the contact area between the metal surface and corrosive media, leading to a decreased corrosion rate. Additionally, the incorporation of nanomaterials, exemplified by SiO2@CuO core-shell nanoparticles, improves the adhesion and durability of the coatings on aluminum alloy substrates. Experimental data from Tafel curve analysis and electrochemical impedance spectroscopy (EIS) confirm the superior corrosion resistance of the superhydrophobic modified aluminum alloy surface compared to untreated surfaces. Estimations indicate a significant reduction in corrosion rate after superhydrophobic treatment. Furthermore, an optical absorption spectra analysis of the core-shell nanoparticles demonstrates their suitability for photocatalytic applications, showcasing their potential contribution to enhancing the overall performance of the coated surfaces. This research underscores the promising approach of combining superhydrophobic properties with photocatalytic capabilities to develop advanced surface modification techniques for enhanced corrosion resistance and functional properties in diverse industrial settings.

Automatic high-throughput and non-invasive selection of sperm at the biochemical level.

BACKGROUND: Sperm selection, a key step in assisted reproductive technology (ART), has long been restrained at the preliminary physical level (morphology or motility); however, subsequent fertilization and embryogenesis are complicated biochemical processes. Such an enormous "gap" poses tough problems for couples dealing with infertility, especially patients with severe/total asthenozoospermia . METHODS: We developed a biochemical-level, automatic-screening/separation, smart droplet-TO-hydrogel chip (BLASTO-chip) for sperm selection. The droplet can sense the pH change caused by sperm's respiration products and then transforms into a hydrogel to be selected out. FINDINGS: The BLASTO-chip system can select biochemically active sperm with an accuracy of over 90%, and its selection efficiency can be flexibly tuned by nearly 10-fold. All the substances in the system were proven to be biosafe via evaluating mice fertilization and offspring health. Live sperm down to 1% could be enriched by over 76-fold to 76%. For clinical application to patients with severe/total asthenozoospermia, the BLASTO-chip could select live sperm from human semen samples containing 10% live but 100% immotile sperm. The rates of fertilization, cleavage, early embryos, and blastocysts were drastically elevated from 15% to 70.83%, 10% to 62.5%, 5% to 37.5%, and 0% to 16.67%, respectively. CONCLUSIONS: The BLASTO-chip represents a real biochemical-level technology for sperm selection that is completely independent of sperm's motility. It can be a powerful tool in ART, especially for patients with severe/total asthenozoospermia. FUNDING: This work was funded by the Ministry of Science and Technology of China, the Ministry of Education of China, and the Shenzhen-Hong Kong Hetao Cooperation Zone.

Platelet Membrane-Coated r-SAK Improves Thrombolytic Efficacy by Targeting Thrombus.

Thrombolytic therapy is one of the most effective treatments for thrombus dissolution and recanalization of blocked vessels in thrombotic diseases. However, the application of the thrombolytic strategy has been limited due to unsatisfactory thrombolytic efficacy, relatively higher bleeding complications, and consequently restricted indications. Recombinant staphylokinase (r-SAK) is a third-generation thrombolytic agent produced by genetic engineering technology, which exhibits a better thrombolytic efficacy than urokinase and recombinant streptokinase. Inspired by the natural affinity of platelets in hemostasis and pathological thrombosis, we developed a platelet membrane (PM)-coated r-SAK (PM-r-SAK). Results from animal experiments and human in vitro studies showed that the PM-r-SAK had a thrombolytic efficacy equal to or better than its 4-fold dose of r-SAK. In a totally occluded rabbit femoral artery thrombosis model, the PM-r-SAK significantly shortened the initial recanalization time compared to the same dose and 4-fold dose of r-SAK. Regarding the recanalized vessels, the PM-r-SAK prolonged the time of reperfusion compared to the same dose and 4-fold dose of r-SAK, though the differences were not significant. An in vitro thrombolytic experiment demonstrated that the thrombolytic efficacy of PM-r-SAK could be inhibited by platelet-poor plasma from patients taking aspirin and ticagrelor. PM coating significantly improves the thrombolytic efficacy of r-SAK, which is related to the thrombus-targeting activity of the PM-r-SAK and can be inhibited by aspirin- and ticagrelor-treated plasma.

A hybrid simulator with novel bicone-wire grid antenna.

The simulator is an indispensable device for the study of high-altitude electromagnetic pulse (HEMP) effects. In order to improve the quality of the electric field waveforms of the HEMP simulator, we carried out simulation and experimental studies for the biconical-wire grid antenna. A new bicone-wire grid antenna HEMP simulator was designed by using skeletonized cones instead of solid cones, which can substantially improve the mobility of the simulator. The effects of different aspect ratios and different cone structures of the simulator on the waveform and distribution characteristics of the electric field were analyzed. The accuracy of the simulation results was verified by testing the bicone-wire grid antenna simulator with the new structure. The results obtained are as follows: First, the simulator can provide a HEMP environment compliant with the International Electrotechnical Commission (IEC) standard in an area of 5 × 6 × 2 m3. Second, the simulator possesses optimal waveform quality and working space among simulators with different aspect ratios. Finally, the influence of using skeletonized cones instead of solid cones on the simulator's electric field waveforms is negligible, and the mobility of the new simulator is significantly improved compared to the original simulator.

Nomogram for predicting moderate-to-severe sleep-disordered breathing in patients with acute ischaemic stroke: a retrospective cohort study.

OBJECTIVES: Moderate-to-severe sleep-disordered breathing (SDB) is prevalent in patients with acute ischaemic stroke (AIS) and is associated with an increased risk of unfavourable prognosis. We aimed to develop and validate a reliable scoring system for the early screening of moderate-to-severe SDB in patients with AIS, with the objective of improving the management of those patients at risk. STUDY DESIGN: We developed and validated a nomogram model based on univariate and multivariate logistic analyses to identify moderate-to-severe SDB in AIS patients. Moderate-to-severe SDB was defined as an apnoea-hypopnoea index (AHI) ≥15. To evaluate the effectiveness of our nomogram, we conducted a comparison with the STOP-Bang questionnaire by analysing the area under the receiver operating characteristic curve. SETTING: Large stroke centre in northern Shanghai serving over 4000 inpatients, 100 000 outpatients and emergency visits annually. PARTICIPANTS: We consecutively enrolled 116 patients with AIS from the Shanghai Tenth People's Hospital. RESULTS: Five variables were independently associated with moderate-to-severe SDB in AIS patients: National Institutes of Health Stroke Scale score (OR=1.20; 95% CI 0.98 to 1.47), neck circumference (OR=1.50; 95% CI 1.16 to 1.95), presence of wake-up stroke (OR=21.91; 95% CI 3.08 to 156.05), neuron-specific enolase level (OR=1.27; 95% CI 1.05 to 1.53) and presence of brainstem infarction (OR=4.21; 95% CI 1.23 to 14.40). We developed a nomogram model comprising these five variables. The C-index was 0.872, indicated an optimal agreement between the observed and predicted SDB patients. CONCLUSIONS: Our nomogram offers a practical approach for early detection of moderate-to-severe SDB in AIS patients. This tool enables individualised assessment and management, potentially leading to favourable outcomes.

Heterocycle compounds synthesized by amide ligand-promoted copper salt catalyzed construction of C-O(S) bonds.

We introduce a mild method for the ligand-promoted copper-catalyzed coupling of 2-halophenol to construct DBDO using cost-effective copper salts, ligands, and alkaline reagents. This method cleverly makes 2-bromophenol complete the Ullman reaction twice, achieves efficient C-O(S) bond coupling and intermolecular cyclization, and yields high amounts of oxygen(sulfur)-containing six-membered ring products. Less reactive 2-chlorophenol was also applied in this catalytic system. The application range of the copper-amide catalytic system was further expanded. Moreover, the success of a gram-scale reaction demonstrated that this operationally simple process is scalable.

The Application of Bilayer Heterogeneous MOFs in pH and Heat-Triggered Systems for Controllable Fragrance Release.

To facilitate the integration of a fragrance encapsulation system into different products to achieve effective releases, a dual-responsive release system with pH and thermal trigger control is designed in this work. A series of ZIF-8 (M) and bilayer ZIF-8-on-ZIF-8 (MM) materials are synthesized by a solvent method at room temperature. The fragrance is encapsulated into the ZIFs by dynamic adsorption or in situ encapsulation combined dynamic adsorption. The fragrance loading contributed by dynamic adsorption was as high as 80%. The fragrance loaded in the double-layer MM host was almost twice that of the monolayer host M due to the stronger electrostatic interaction between MM and vanillin. In the pH and thermal trigger response release experiments, the second MOF layer in the MM host, as a controlled entity, greatly improved the load and kinetic equilibrium time of vanillin, and realized the controlled release of guest molecules. The developed dual-responsive release system in this work exhibits great potential in daily chemical products.

InCl3/TfOH-Mediated Convenient Synthesis of 3-Alkylideneoxindoles from 2-Oxindoles with 1,3-Diones, Ketones, or Aldehydes.

Two efficient and convenient methods for the synthesis of 3-alkylideneoxindoles are described in this paper. The InCl3/TfOH-mediated tandem Knoevenagel condensation-deacylation sequence of various 2-oxindoles with 1,3-diones or acetoacetate furnished 3-alkylideneoxindoles in satisfactory to excellent yields (up to >99% yield). Employing the reaction system, the condensation of 2-oxindoles with ketones or aldehydes also proceeded smoothly to produce 3-alkylideneoxindoles. This protocol can be amenable to scale up. The effect of acids on this condensation reaction and intermolecular competition experiments were investigated to understand the aspect of the reaction.

Fathers' needs of breastfeeding support: Perspective of health nurses.

PURPOSE: To explore the improvement of health education on father's participation in breastfeeding from the perspective of maternal and child health nurses. METHODS: Qualitative phenomenological research was used, and 15 maternal and child health nurses who provided breastfeeding support were invited. With semi-structured deep interviews and on-site recordings, data were analyzed through content analysis. RESULTS: Four main themes were extracted, including 'cultivating fathers' awareness of participation in breastfeeding', 'collaboration of multiple disciplines to improve health education on breastfeeding for fathers in hospital', 'Simulated scenarios to develop fathers' skills in solving breastfeeding problems', and 'establishing a hospital-community interface network to improve breastfeeding continuation care after hospital discharge'. CONCLUSIONS: Medical and health care departments should attach importance to guidance on health education for fathers' breastfeeding participation, cultivate fathers' awareness of participation in breastfeeding, provide multi-disciplinary collaboration-based health education on breastfeeding for fathers from the prenatal period and improve post-discharge health education on breastfeeding. The additional education being suggested would contribute to fathers being able to play an important role in breastfeeding.

Therapeutic role of miR-26a on cardiaorenal injury in mice model of angiotensin-II induced chronic kidney disease through inhibition of LIMS1/ILK pathway.

BACKGROUND: Chronic kidney disease (CKD) is associated with common pathophysiological processes, such as inflammation and fibrosis, in both the heart and the kidney. However, the underlying molecular mechanisms that drive these processes are not yet fully understood. Therefore, this study focused on the molecular mechanism of heart and kidney injury in CKD. METHODS: We generated a microRNA (miR)-26a knockout (KO) mouse model to investigate the role of miR-26a in angiotensin (Ang)-II-induced cardiac and renal injury. We performed Ang-II modeling in wild type (WT) mice and miR-26a KO mice, with six mice in each group. In addition, Ang-II-treated AC16 cells and HK2 cells were used as in vitro models of cardiac and renal injury in the context of CKD. Histological staining, immunohistochemistry, quantitative real-time polymerase chain reaction (PCR), and Western blotting were applied to study the regulation of miR-26a on Ang-II-induced cardiac and renal injury. Immunofluorescence reporter assays were used to detect downstream genes of miR-26a, and immunoprecipitation was employed to identify the interacting protein of LIM and senescent cell antigen-like domain 1 (LIMS1). We also used an adeno-associated virus (AAV) to supplement LIMS1 and explored the specific regulatory mechanism of miR-26a on Ang-II-induced cardiac and renal injury. Dunnett's multiple comparison and t-test were used to analyze the data. RESULTS: Compared with the control mice, miR-26a expression was significantly downregulated in both the kidney and the heart after Ang-II infusion. Our study identified LIMS1 as a novel target gene of miR-26a in both heart and kidney tissues. Downregulation of miR-26a activated the LIMS1/integrin-linked kinase (ILK) signaling pathway in the heart and kidney, which represents a common molecular mechanism underlying inflammation and fibrosis in heart and kidney tissues during CKD. Furthermore, knockout of miR-26a worsened inflammation and fibrosis in the heart and kidney by inhibiting the LIMS1/ILK signaling pathway; on the contrary, supplementation with exogenous miR-26a reversed all these changes. CONCLUSIONS: Our findings suggest that miR-26a could be a promising therapeutic target for the treatment of cardiorenal injury in CKD. This is attributed to its ability to regulate the LIMS1/ILK signaling pathway, which represents a common molecular mechanism in both heart and kidney tissues.

Self-supervised pre-training for joint optic disc and cup segmentation via attention-aware network.

Image segmentation is a fundamental task in deep learning, which is able to analyse the essence of the images for further development. However, for the supervised learning segmentation method, collecting pixel-level labels is very time-consuming and labour-intensive. In the medical image processing area for optic disc and cup segmentation, we consider there are two challenging problems that remain unsolved. One is how to design an efficient network to capture the global field of the medical image and execute fast in real applications. The other is how to train the deep segmentation network using a few training data due to some medical privacy issues. In this paper, to conquer such issues, we first design a novel attention-aware segmentation model equipped with the multi-scale attention module in the pyramid structure-like encoder-decoder network, which can efficiently learn the global semantics and the long-range dependencies of the input images. Furthermore, we also inject the prior knowledge that the optic cup lies inside the optic disc by a novel loss function. Then, we propose a self-supervised contrastive learning method for optic disc and cup segmentation. The unsupervised feature representation is learned by matching an encoded query to a dictionary of encoded keys using a contrastive technique. Finetuning the pre-trained model using the proposed loss function can help achieve good performance for the task. To validate the effectiveness of the proposed method, extensive systemic evaluations on different public challenging optic disc and cup benchmarks, including DRISHTI-GS and REFUGE datasets demonstrate the superiority of the proposed method, which can achieve new state-of-the-art performance approaching 0.9801 and 0.9087 F1 score respectively while gaining 0.9657 D C disc and 0.8976 D C cup . The code will be made publicly available.

Association of adherence to the Chinese version of the MIND diet with reduced cognitive decline in older Chinese individuals: Analysis of the Chinese Longitudinal Healthy Longevity Survey.

BACKGROUND: Current evidence suggests that the Mediterranean-Dietary Approaches to Stop Hypertension (DASH) diet intervention for Neurodegenerative Delay (MIND) is associated with a reduced risk of cognitive impairment among North American and Oceanian populations. However, there has been limited exploration of whether this association extends to the Asian population. This study aimed to assess the correlation between the Chinese version of the MIND (cMIND) diet and cognitive impairment in older Chinese individuals. METHODS: We utilized data from the 2008 wave of the Chinese Longitudinal Healthy Longevity Survey. Participants aged ≥65 years with normal cognitive function at baseline were enrolled. The cMIND diet score (cMINDDS) was calculated by assessing dietary patterns based on survey responses. The Chinese version of the Mini-Mental State Examination (MMSE) was employed to diagnose cognitive impairment in participants. We stratified the analysis by cMINDDS and conducted additional sensitivity analyses. RESULTS: The cohort consisted of 6411 participants. Over a 3-year follow-up, 1165 (18.6%) individuals who initially had normal cognitive function developed cognitive impairment. A linear association was observed between cMINDDS and cognitive impairment. The increased cMINDDS was associated with a reduced risk of cognitive impairment (quartile 1 vs. quartile 4: the adjusted odds ratio [OR] = 0.77, 95% confidence interval [CI]: [0.60, 0.97], p trend = 0.023). Regarding food composition, higher consumption of fresh fruits and nuts was associated with a decreased risk of cognitive impairment (OR = 0.77, 95% CI: [0.66, 0.89] and OR = 0.70, 95% CI [0.58, 0.86], respectively). CONCLUSIONS: Adherence to the cMIND diet was associated with lower risks of cognitive impairment in older Chinese individuals. The cMIND diet, based on the MIND dietary pattern, could serve as a preventive measure against cognitive impairment.

Whole-exome sequencing has revealed novel genetic characteristics in intracranial germ cell tumours in the Chinese.

AIMS: Intracranial germ cell tumour (IGCT) is a type of rare central nervous system tumour that mainly occurs in children and adolescents, with great variation in its incidence rate and molecular characteristics in patients from different populations. The genetic alterations of IGCT in the Chinese population are still unknown. METHODS AND RESULTS: In this study, 47 patients were enrolled and their tumour specimens were analysed by whole-exome sequencing (WES). We found that KIT was the most significantly mutated gene (15/47, 32%), which mainly occurred in the germinoma group (13/20, 65%), and less frequently in NGGCT (2/27, 7%). Copy number variations (CNVs) of FGF6 and TFE3 only appeared in NGGCT patients (P = 0.003 and 0.032, respectively), while CNVs of CXCR4, RAC2, PDGFA, and FEV only appeared in germinoma patients (P = 0.004 of CXCR4 and P = 0.027 for the last three genes). Compared with a previous Japanese cohort, the somatic mutation rates of RELN and SYNE1 were higher in the Chinese. Prognostic analysis showed that the NF1 mutation was associated with shorter overall survival and progression-free survival in IGCT patients. Clonal evolution analysis revealed an early branched evolutionary pattern in two IGCT patients who underwent changes in the histological subtype or degree of differentiation during disease surveillance. CONCLUSION: This study indicated that Chinese IGCT patients may have distinct genetic characteristics and identified several possible genetic alterations that have the potential to become prognostic biomarkers of NGGCT patients.