Stable inhibition of choroidal neovascularization by adeno-associated virus 2/8-vectored bispecific molecules.

Neovascular age-related macular degeneration (nAMD) causes severe visual impairment. Pigment epithelium-derived factor (PEDF), soluble CD59 (sCD59), and soluble fms-like tyrosine kinase-1 (sFLT-1) are potential therapeutic agents for nAMD, which target angiogenesis and the complement system. Using the AAV2/8 vector, two bi-target gene therapy agents, AAV2/8-PEDF-P2A-sCD59 and AAV2/8-sFLT-1-P2A-sCD59, were generated, and their therapeutic efficacy was investigated in laser-induced choroidal neovascularization (CNV) and Vldlr-/- mouse models. After a single injection, AAV2/8-mediated gene expression was maintained at high levels in the retina for two months. Both AAV2/8-PEDF-P2A-sCD59 and AAV2/8-sFLT-1-P2A-sCD59 significantly reduced CNV development for an extended period without side effects and provided efficacy similar to two injections of current anti-vascular endothelial growth factor monotherapy. Mechanistically, these agents suppressed the extracellular signal-regulated kinase and nuclear factor-κB pathways, resulting in anti-angiogenic activity. This study demonstrated the safety and long-lasting effects of AAV2/8-PEDF-P2A-sCD59 and AAV2/8-sFLT-1-P2A-sCD59 in CNV treatment, providing a promising therapeutic strategy for nAMD.

Fbxo28 is essential for spindle migration and morphology during mouse oocyte meiosis I.

Spindle migration and assembly regulates asymmetric oocyte division, which is essential for fertility. Fbxo28, as a member of SCF (Skp1-Cul1-F-box) ubiquitin E3 ligases complex, is specifically expressed in oocytes. However, little is known about the functions of Fbxo28 in spindle assembly and migration during oocyte meiosis I. In present study, microinjection with morpholino oligonucleotides and exogenous mRNA for knockdown and rescue experiments, and immunofluorescence staining, western blot, timelapse confocal microscopy and chromosome spreading were utilized to explore the roles of Fbxo28 in asymmetric division during meiotic maturation. Our data suggested that Fbxo28 mainly localized at chromosomes and acentriolar microtubule-organizing centers (aMTOCs). Depletion of Fbxo28 did not affect polar body extrusion but caused defects in spindle morphology and migration, indicative of the failure of asymmetric division. Moreover, absence of Fbxo28 disrupted both cortical and cytoplasmic actin assembly and decreased the expression of ARPC2 and ARP3. These defects could be rescued by exogenous Fbxo28-myc mRNA supplement. Collectively, this study demonstrated that Fbxo28 affects spindle morphology and actin-based spindle migration during mouse oocyte meiotic maturation.

One-step synthesized multisize AuAg alloy nanoparticles with high SERS sensitivity in directly detecting SARS-CoV-2 spike protein.

The coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has resulted in widespread disease transmission, challenging the stability of global healthcare systems. Surface-enhanced Raman scattering (SERS) as an easy operation, fast, and low-cost technology illustrates a good potential in detecting SARS-CoV-2. In the study, one-step fabrication of gold-silver alloy nanoparticles (AuAgNPs) with adjustable metal proportions and diameters is employed as SERS substrates. The angiotensin-converting enzyme 2 (ACE2) functionalized AuAgNPs are applied as sensor surfaces to detect SARS-CoV-2 S protein. By optimizing the SERS substrates, ACE2/Au35Ag65NPs illustrate higher performance in detecting the SARS-CoV-2 S protein with a limit of detection (LOD) of 10 fg/mL in both phosphate-buffered saline (PBS) and pharyngeal swabs solution (PSS). It also provides excellent reproducibility with a relative standard deviation (RSD) of 7.7 % and 7.9 %, respectively. This easily preparable and highly reproducible SERS substrate has good potential in the practical application of detecting SARS-CoV-2.

The Three-Dimensional Hierarchical Structure of Bi x -Sn1-x O2 Used for Ultrasensitive Detection of Butanone under UV Irradiation.

Recent studies have identified butanone as a promising biomarker in the breath of lung cancer patients, yet the understanding of its gas-sensing properties remains limited. A key challenge has been to enhance the gas-sensing performance of materials toward butanone, particularly under ultraviolet light exposure. Herein, we report the synthesis of a novel three-dimensional composite material composed of SnO2 incorporated with Bi2O3 using facile hydrothermal and impregnation precipitation methods. Detailed physical and chemical characterizations were performed to assess the properties of the developed material. Upon activation with ultraviolet light, our composite exhibited exceptionally high sensitivity to butanone. Remarkably, the butanone response was nearly 3 times greater for the Bi2O3-loaded SnO2 composite than for pristine SnO2, achieving a response value of 70. This substantial improvement is due to the synergistic effect of the material's distinctive three-dimensional architecture and the presence of Bi2O3, which significantly augmented the gas-sensing capability of butanone. To elucidate the underlying gas-sensing mechanism, we conducted first-principles calculations using density functional theory (DFT). The computational analysis revealed that the Bi2O3-containing system possesses superior adsorption energy for butanone. Ultimately, our findings suggest that the Bi-SnO2 composite holds great promise as an optimal sensing material for the detection of butanone under ultraviolet illumination.

TransMVAN: Multi-view Aggregation Network with Transformer for Pneumonia Diagnosis.

Automated and accurate classification of pneumonia plays a crucial role in improving the performance of computer-aided diagnosis systems for chest X-ray images. Nevertheless, it is a challenging task due to the difficulty of learning the complex structure information of lung abnormality from chest X-ray images. In this paper, we propose a multi-view aggregation network with Transformer (TransMVAN) for pneumonia classification in chest X-ray images. Specifically, we propose to incorporate the knowledge from glance and focus views to enrich the feature representation of lung abnormality. Moreover, to capture the complex relationships among different lung regions, we propose a bi-directional multi-scale vision Transformer (biMSVT), with which the informative messages between different lung regions are propagated through two directions. In addition, we also propose a gated multi-view aggregation (GMVA) to adaptively select the feature information from glance and focus views for further performance enhancement of pneumonia diagnosis. Our proposed method achieves AUCs of 0.9645 and 0.9550 for pneumonia classification on two different chest X-ray image datasets. In addition, it achieves an AUC of 0.9761 for evaluating positive and negative polymerase chain reaction (PCR). Furthermore, our proposed method also attains an AUC of 0.9741 for classifying non-COVID-19 pneumonia, COVID-19 pneumonia, and normal cases. Experimental results demonstrate the effectiveness of our method over other methods used for comparison in pneumonia diagnosis from chest X-ray images.

AML cell-derived exosomes suppress the activation and cytotoxicity of NK cells in AML via PD-1/PD-L1 pathway.

Exosomes are bilayer lipid bodies and contain a variety of bioactive molecules such as proteins, lipids, and nucleic acids, and so forth. Exosomes derived from solid tumors may play critical roles in tumor development and immune evasion. However, the underlying effects of tumor-derived exosomes on immune function in modulating intercellular crosstalk within the bone marrow niche during acute myeloid leukemia (AML) development and immune evasion remain largely elusive. In this study, we aimed to explore the role of AML-exos in AML immune evasion. First, we isolated tumor-derived exosomes from AML cells (AML-exos) and revealed the presence of programmed cell death ligand-1 (PD-L1) protein in AML-exos. Next, we demonstrated that AML-exos can directly suppress the activation of natural killer (NK) cells and inhibit the cytotoxicity of NK cells, probably through activating the programmed cell death-1 (PD-1)/PD-L1 pathway. Furthermore, the inhibitory effect of AML-exos on NK cells could be alleviated by either PD-L1 inhibitor or antagonist. In summary, we demonstrated that AML-exos possess a PD-L1-dependent tumor-promoting effect which may contribute to immune tolerance in antitumor therapy, but blocking the PD-1/PD-L1 pathway may alleviate the tumor immunosuppression induced by AML-exos. Our findings in this study may offer a new immunotherapy strategy to cure AML.

Evaluating the advancements in a recently introduced universal adhesive compared to its predecessor.

Background/purpose: The dental adhesive market is constantly evolving to meet the demands of dentists and patients, but new products and upgrades should be rigorously evaluated before being used in clinical practice. This study investigated the physicomechanical properties and dentin bonding efficacy of a newly upgraded universal adhesive compared to its predecessor. Materials and methods: Twenty-four molars were divided into four groups (n = 6/group) based on adhesive (new vs. predecessor) and application mode [self-etch (SE) vs. etch-and-rinse (ER)] for evaluating their dentin microtensile bond strength (µTBS), failure pattern, and bonding interface. Additional thirty-six molars' crowns were perpendicularly sectioned to obtain flat mid-coronal dentin discs. The opposing dentin surfaces of each disc received contrasting treatments (new/predecessor adhesive applied in SE/ER mode), resulting in six interventions. The bonded discs (n = 6/intervention) were used to assess the adhesives' survival probability employing a double-sided µTBS test. The other physicomechanical properties examined were adhesives' oxygen inhibition layer (OIL), viscosity, hardness, elastic modulus, degree of conversion (DC), and in-situ DC. Results: Both adhesive versions showed similar µTBS (P > 0.05), failure pattern (P > 0.05), and survival probability (P > 0.008). ER mode promoted resin tag formation and exhibited a slender adhesive layer for both adhesives. The newer adhesive version showed a thinner adhesive layer in general with narrower OIL (P < 0.001), less viscosity (P < 0.001), higher hardness (P < 0.05), elastic modulus (P < 0.05), DC (P < 0.001), and in-situ DC (P < 0.001). Conclusion: While the newly updated adhesive had superior physicomechanical properties with more fluidity, its dentin bonding efficacy and survival probability were comparable to its predecessor.

The therapeutic effect and metabolic mechanism analysis of Guilingji on idiopathic oligo-asthenoteratozoospermia.

Introduction: Guilingji, a famous traditional Chinese medicine (TCM) formula, has been used to combat aging and male sexual dysfunction in China for centuries. To date, there has been little evidence-based clinical research on the use of Guilingji to treat idiopathic oligo-asthenoteratozoospermia (OAT), and the therapeutic mechanism from a metabolic perspective needs to be investigated further. Methods: This was a multicenter, double-blind, randomized controlled clinical study of 240 patients with idiopathic OAT recruited from four hospitals between January 2020 and January 2022. Patients were randomly assigned in a 1꞉1 ratio to receive oral Guilingji capsules or placebo for 12 weeks. The total progressive motile sperm count (TPMSC) was considered the primary outcome, and the other sperm parameters, seminal plasma parameters and serum hormones were considered the secondary outcome. A nontargeted metabolomics analysis of serum from OAT patients before and after Guilingji administration was performed by HPLC-MS to identify key metabolites. Furthermore, we used a rat model to show spermatogenesis phenotypes to validate the effect of the key metabolites screened from the patients. Results: At weeks 4, 8 and 12, TPMSC and other sperm parameters were significantly improved in the Guilingji group compared with the placebo group (P < 0.05 for all comparisons). At week 4, superoxide dismutase (SOD) and acrosomal enzyme activity of seminal plasma were significantly elevated in the Guilingji group compared with the placebo group, while reactive oxygen species (ROS) levels were significantly reduced (P < 0.05). Lactate dehydrogenase-X (LDHX) levels appeared to be significantly increased after 12 weeks continuous medication compared with Placebo group (P = 0.032). The metabolomics analysis of serum from OAT patients before and after Guilingji administration showed that the glucose-6-phosphate (G6P) concentration in patients' serum was significantly elevated after Guilingji treatment. Compared to the control, when Kidney-Yang deficiency model rats were treated with Guilingji or its key intermediate metabolite G6P, their sperm concentration and spermatozoic activity were improved similarly, and their structural damage of rat's testicular and epididymal tissues were recovered. Conclusion: This study provided valuable clinical evidence for the utility of Guilingji as a treatment for OAT. These findings thus demonstrate that G6P is involved in the therapeutic mechanism of Guilingji in OAT treatment based on clinical and rat intervention studies.

Fluorescent probe for imaging intercellular tension: molecular force approach.

Cellular mechanical force plays a crucial role in numerous biological processes, including wound healing, cell development, and metastasis. To enable imaging of intercellular tension, molecular tension probes were designed, which offer a simple and efficient method for preparing Au-DNA intercellular tension probes with universal applicability. The proposed approach utilizes gold nanoparticles linked to DNA hairpins, enabling sensitive visualization of cellular force in vitro. Specifically, the designed Au-DNA intercellular tension probe includes a molecular spring flanked by a fluorophore-quencher pair, which is anchored between cells. As intercellular forces open the hairpin, the fluorophore is de-quenched, allowing for visualization of cellular force. The effectiveness of this approach was demonstrated by imaging the cellular force in living cells using the designed Au-DNA intercellular tension probe.

Formation of Stable Vascular Networks by 3D Coaxial Printing and Schiff-Based Reaction.

Vascularized organs hold potential for various applications, such as organ transplantation, drug screening, and pathological model establishment. Nevertheless, the in vitro construction of such organs encounters many challenges, including the incorporation of intricate vascular networks, the regulation of blood vessel connectivity, and the degree of endothelialization within the inner cavities. Natural polymeric hydrogels, such as gelatin and alginate, have been widely used in three-dimensional (3D) bioprinting since 2005. However, a significant disparity exists between the mechanical properties of the hydrogel materials and those of human soft tissues, necessitating the enhancement of their mechanical properties through modifications or crosslinking. In this study, we aim to enhance the structural stability of gelatin-alginate hydrogels by crosslinking gelatin molecules with oxidized pullulan (i.e., a polysaccharide) and alginate molecules with calcium chloride (CaCl2). A continuous small-diameter vascular network with an average outer diameter of 1 mm and an endothelialized inner surface is constructed by printing the cell-laden hydrogels as bioinks using a coaxial 3D bioprinter. The findings demonstrate that the single oxidized pullulan crosslinked gelatin and oxidized pullulan/CaCl2 double-crosslinked gelatin-alginate hydrogels both exhibit a superior structural stability compared to their origins and CaCl2 solely crosslinked gelatin-alginate hydrogels. Moreover, the innovative gelatin and gelatin-alginate hydrogels, which have excellent biocompatibilities and very low prices compared with other hydrogels, can be used directly for tissue/organ construction, tissue/organ repairment, and cell/drug transportation.

Tailoring the Ni-O Microenvironment in Amorphous-Dominated Highly Active and Stable Zn/NiO for Hydrogen Sulfide Detection.

Amorphous metal oxide semiconductor (MOS) materials are endowed with great promise to modulate electronic structures for gas-sensing performance improvement. However, the elevated-temperature requirement of gas sensors severely impedes the application of amorphous materials due to their low thermal stability. Here, a cationic-assisted strategy to tailor the Ni-O microenvironment in an amorphous-dominated Zn/NiO heterogeneous structure with high thermal stability was developed. It was found that 6 mol % Zn incorporation into amorphous NiO can effectively preserve the amorphous-dominated NiO phase even at high temperature. After calcination, the amorphous oxide can only be converted to crystals partly thus leading to the formation of amorphous/crystalline compounds, and the content of the amorphous phase can be adjusted by changing the calcination temperature. This amorphous/crystalline configuration can induce more electron transfer from Ni to Zn species, leading to the formation of active Niδ+ (δ>2) centers. Ex situ XPS and in situ Raman spectroscopy studies proved that the generated Niδ+ species pronouncedly promote the electron transfer during the H2S adsorption process. The amorphous/crystalline-6 mol % Zn/NiO sensor exhibits exceptional hydrogen sulfide response (2 ppm, 3.23), outstanding repeatability (as long as 5 weeks), and low limit of detection (as low as 50 ppb), surpassing most reported nickel-based gas sensors such as the crystal nickel oxide prepared in this work. The response and detection limit of the latter is only (2 ppm, 1.89) and (0.05 ppm) respectively. Our work thus opens up more opportunities for fundamental understanding and modulating of highly active amorphous sensing materials.

Acid-Promoted Multicomponent Reaction To Synthesize 4-Phosphorylated 4H-Chromenes.

An effective multicomponent reaction for the synthesis of 4-phosphorylated 4H-chromenes via a tandem phosphorylation/alkylation/cyclization/dehydration sequence with water as the only byproduct was developed. Extensive mechanistic investigations involving in situ NMR experiments, time control experiments, and in situ HRMS experiment allowed us to elucidate the order of each subreaction to arrive at a complete understanding of the underlying mechanism of this multicomponent reaction. Mechanistic data confirm that the reaction begins with a phospha-aldol-elimination, followed by addition of a ketone enolate, intermolecular alkylation, intramolecular cyclization, and dehydration under acidic conditions.

Macrophage polarization as a novel endpoint for assessing combined risk of phthalate esters.

Combined exposure to phthalate esters (PAEs) has garnered increasing attention due to potential synergistic effects on human health. This study aimed to develop an in vitro model using human macrophages to evaluate the combined toxicity of PAEs and explore the underlying mechanisms. A high-throughput screening system was engineered by expressing a PPRE-eGFP reporter in THP-1 monocytes to monitor macrophage polarization upon PAEs exposure. Individual PAEs exhibited varied inhibitory effects on M2 macrophage polarization, with mono(2-ethylhexyl) phthalate (MEHP) being the most potent. Isobologram analysis revealed additive interactions when MEHP was combined with other PAEs, resulting in more pronounced suppression of M2 markers compared to individual compounds. Mechanistic studies suggested PAEs may exert effects by modulating PPARγ activity to inhibit M2 polarization. Notably, an equimolar mixture of six PAEs showed additive inhibition of M2 markers. In vivo experiments corroborated the combined hepatotoxic effects, with mice exposed to a PAEs mixture exhibiting reduced liver weight, dyslipidemia, and decreased hepatic M2 macrophages compared to DEHP alone. Transcriptome analysis highlighted disruptions in PPAR signaling, and distinct pathway alterations on cholesterol metabolism in the mixture group. Collectively, these findings underscore the importance of evaluating mixture effects and provide a novel approach for hazard assessment of combined PAEs exposure with implications for environmental health risk assessment.

An antifouling supramolecular polymer ophthalmic ointment alleviates symblepharon in rat alkali burn eyes.

Symblepharon is an adverse ocular disease resulting in ocular discomfort and impaired vision, severely dragging down a patient's quality of life. Due to the specificity of the ocular surface, the retention time of drugs on it is short, leading to limited therapeutic effects for ocular diseases. Therefore, it is imperative to design a novel drug delivery system, which can not only prolong the retention time of a drug but also play an anti-fibrosis role in symblepharon. Herein, an antifouling supramolecular polymer ophthalmic ointment consisting of poly(N-acryloyl alaninamide) (PNAAA), vitamin C (VitC) and levofloxacin (Levo) was developed (termed PNAVL ophthalmic ointment), which acted as a mucoadhesive and long-acting ocular delivery system. This antifouling PNAVL ophthalmic ointment improved the retention time of VitC and Levo, and simultaneously provided anti-inflammation and anti-fibrosis effects for mitigating symblepharon after ocular alkali burn injury.

Quantitative analysis of cement raw materials based on nanoparticle-enhanced laser-induced breakdown spectroscopy.

The nanoparticle-enhanced laser-induced breakdown spectroscopy (NELIBS) technique has attracted much attention because of its significant spectral enhancement as well as the reduction of spectral noise. More NELIBS studies have focused on the effect of nanoparticles on spectral intensity and the optimization of experiments. NELIBS has not been studied for the detection of cement raw material components and the repeatability of quantitative analysis. In this paper, the effect of NELIBS on the spectral quality as well as the repeatability of the quantitative analysis of cement raw materials is investigated. The effects of different AuNP sizes and volumes on LIBS brought about by pre-ablation were compared and modeled for quantitative analysis. The results showed that the signal-to-noise ratio (SNR) of NELIBS spectra after pre-ablation was improved from 2.72 to 7.81, and the relative standard deviations (RSDs) of CaO, SiO2, Al2O3, and Fe2O3 in cement raw materials were reduced by 47%, 30%, 31%, and 33%, respectively. In summary, this paper provides a comprehensive discussion and comparison of AuNPs versus LIBS for quantitative analysis of cement raw material components, and also provides a new solution for quantitative analysis of cement raw materials.

Organic Reactions Enabled by Mechanical Force-Induced Single Electron Transfer.

Mechanochemical reactions, achieved through milling, grinding, or other mechanical actions, have emerged as a solvent-free alternative to traditional solution-based chemistry. Mechanochemistry not only provides the opportunity to eliminate bulk solvent use, reducing waste generation, but also unveils a new reaction strategy which enables the realization of reactions previously inaccessible in solution. While the majority of organic reactions facilitated by mechanical force traditionally follow two-electron transfer pathways similar to their solution-based counterparts, the field of mechanochemically induced single-electron transfer (SET) reactions has witnessed rapid development. This review outlines examples of mechanochemical reactions facilitated by the SET process, focusing on the reagents that initiate SET, thereby positioning mechanochemistry as a burgeoning field within the realm of single-electron chemistry.

Europe as a secondary distribution hub in the worldwide invasion of the potato cyst nematode Globodera rostochiensis.

The potato cyst nematode Globodera rostochiensis originates from the Andean Mountain region in South America and has unintentionally been introduced to all inhabited continents. Several studies have examined the population genetic structure of this pest in various countries by using microsatellite markers. However, merging microsatellite data produced from different laboratories is challenging and can introduce uncertainty when interpreting the results. To overcome this challenge and to explore invasion routes of this pest, we have genotyped 22 G. rostochiensis populations from all continents. Within populations, the highest genetic diversity was observed in the South American populations, the European populations showed an intermediate level of genetic diversity and the remaining populations were the less diverse. This confirmed pre-existing knowledge such as a first introduction event from South America to Europe, but the less diverse populations could originate either from South America or from Europe. At the continental scale, STRUCTURE genetic clustering output indicated that North America and Asia have experienced at least two introduction events. Comparing different evolutionary scenarios, the Approximate Bayesian Computation analysis showed that Europe served as a secondary distribution centre for the invasion of G. rostochiensis into all other continents (North America, Africa, Asia and Oceania).

The Effect of Nanoscale Modification of Nisin by Different Milk-Derived Proteins on Its Physicochemical Properties and Antibacterial Activity.

Nisin is used as a natural food preservative because of its broad-spectrum antimicrobial activity against Gram-positive bacteria. However, free nisin is susceptible to various factors that reduce its antimicrobial activity. Milk protein, a protein derived from milk, has self-assembly properties and is a good carrier of bioactive substances. In this study, lactoferrin-nisin nanoparticles (L-N), bovine serum albumin-nisin nanoparticles (B-N), and casein-nisin nanoparticles (C-N) were successfully prepared by a self-assembly technique, and then their properties were investigated. The studies revealed that lactoferrin (LF) and nisin formed L-N mainly through hydrophobic interactions and hydrogen bonding, and L-N had the best performance. The small particle size (29.83 ± 2.42 nm), dense reticular structure, and good thermal stability, storage stability, and emulsification of L-N laid a certain foundation for its application in food. Further bacteriostatic studies showed that L-N enhanced the bacteriostatic activity of nisin, with prominent inhibitory properties against Listeria monocytogenes, Staphylococcus aureus, and Bacillus cereus, which mainly disrupted the cell membrane of the bacteria. The above results broaden our understanding of milk protein-nisin nanoparticles, while the excellent antibacterial activity of L-N makes it promising for application as a novel food preservative, which will help to improve the bioavailability of nisin in food systems.

Longitudinal multicompartment characterization of host-microbiota interactions in patients with acute respiratory failure.

Critical illness can significantly alter the composition and function of the human microbiome, but few studies have examined these changes over time. Here, we conduct a comprehensive analysis of the oral, lung, and gut microbiota in 479 mechanically ventilated patients (223 females, 256 males) with acute respiratory failure. We use advanced DNA sequencing technologies, including Illumina amplicon sequencing (utilizing 16S and ITS rRNA genes for bacteria and fungi, respectively, in all sample types) and Nanopore metagenomics for lung microbiota. Our results reveal a progressive dysbiosis in all three body compartments, characterized by a reduction in microbial diversity, a decrease in beneficial anaerobes, and an increase in pathogens. We find that clinical factors, such as chronic obstructive pulmonary disease, immunosuppression, and antibiotic exposure, are associated with specific patterns of dysbiosis. Interestingly, unsupervised clustering of lung microbiota diversity and composition by 16S independently predicted survival and performed better than traditional clinical and host-response predictors. These observations are validated in two separate cohorts of COVID-19 patients, highlighting the potential of lung microbiota as valuable prognostic biomarkers in critical care. Understanding these microbiome changes during critical illness points to new opportunities for microbiota-targeted precision medicine interventions.