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Abstract Flow cytometry is a key technology for the analysis and sorting of cells or particles at high throughput. Conventional and current flow cytometry is primarily based on fluorescent stains to detect the cells of interest. However, such stains also have disadvantages, as their effect on cells must be carefully tested to avoid effects on the results of the experiments. Alternative approaches using imaging or other label-free techniques often require highly sophisticated setups, are commonly limited in resolution, and produce challenging amounts of data. Our technology exploits scattered light instead. The custom-built flow cytometry setup comprises a fiber array in forward scatter detection for angular resolution and captures the whole pulse shape with advanced signal processing. Thereby this setup enables cell analysis and sorting purely based on scattered light signals without the need for fluorescent labels. We demonstrate the feasibility of this cell sorting technology by sorting cell lines for their cell cycle stages based on scattered light. Furthermore, we demonstrate the ability to classify human peripheral blood T- and B-cell subsets.

ABSTRACTThe prevalence of inflammatory bowel disease (IBD) is rising globally; however, its etiology is still not fully understood. Patient genetics, immune system, and intestinal microbiota are considered critical factors contributing to IBD. Preclinical animal models are crucial to better understand the importance of individual contributing factors. Among these, the dextran sodium sulfate (DSS) colitis model is the most widely used. DSS treatment induces gut inflammation and dysbiosis. However, its exact mode of action remains unclear. To determine whether DSS treatment induces pathogenic changes in the microbiota, we investigated the microbiota-modulating effects of DSS on murine microbiota in vitro. For this purpose, we cultured murine microbiota from the colon in six replicate continuous bioreactors. Three bioreactors were supplemented with 1% DSS and compared with the remaining PBS-treated control bioreactors by means of microbiota taxonomy and functionality. Using metaproteomics, we did not identify significant changes in microbial taxonomy, either at the phylum or genus levels. No differences in the metabolic pathways were observed. Furthermore, the global metabolome and targeted short-chain fatty acid (SCFA) quantification did not reveal any DSS-related changes. DSS had negligible effects on microbial functionality and taxonomy in vitro in the absence of the host environment. Our results underline that the DSS colitis mouse model is a suitable model to study host–microbiota interactions, which may help to understand how intestinal inflammation modulates the microbiota at the taxonomic and functional levels.

We report an elderly male patient with frequent episodes of dizziness due to a complete atrioventricular block who underwent temporary pacemaker insertion in a local hospital. After the implantation of a permanent pacemaker and removal of the temporary pacemaker lead, the patient developed sudden neurological symptoms, upon which an acute cerebellar infarction was diagnosed via head CT. We will discuss the adequacy of the periprocedural administration.

Drought is an important abiotic stressor that adversely affects agricultural production, resulting in decreased crop yields and food insecurity. The objective of this study was to develop a novel hydrogel-based composite (S-B-AA) by incorporating biochar into hydrogel (S-AA) and evaluating its effects on tobacco plants under drought stress. The S-B-AA hydrogel exhibited enhanced water absorption and retention capabilities compared to the S-AA hydrogel. Under drought stress conditions, the S-B-AA hydrogel effectively increased the total water content, relative water content, and free water content of tobacco leaves, while reducing the bound water content and water saturation deficit. Additionally, it improved the stomatal status of tobacco leaves, significantly enhanced the photosynthetic capacity of tobacco seedlings, and promoted biomass accumulation. Moreover, the S-B-AA hydrogel alleviated the excessive accumulation of reactive oxygen species (ROS) induced by drought stress in tobacco seedlings, thereby alleviating ROS damage to cell membranes. It also reduced antioxidant enzyme activity and the levels of the signaling molecule H2S, while minimizing the impact of drought stress on tobacco osmotic regulation. Transcriptome analysis revealed that S-B-AA up-regulated 2826 DEGs and down-regulated 5225 DEGs compared to the control group. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis demonstrated that the differentially expressed genes were predominantly enriched in the mitogen-activated protein kinase (MAPK) signaling pathway, plant hormone signaling pathway, and photosynthesis pathway. This study provides a theoretical basis and practical guidance for the application of S-B-AA hydrogel in agriculture and environmental protection.

Semantic change detection (SCD) extends the traditional change detection (CD) task to simultaneously identify the change areas and their corresponding land cover categories in bi-temporal images. This “from-to” change information holds significant value in numerous practical applications and is increasingly garnering attention in the remote sensing domain. However, prevalent challenges such as loss of detail and class imbalance significantly hinder the efficacy of SCD applications. To address this challenge, we propose a novel semantic information collaboration network (SIC-Net). This network incorporates a detail capture path and a spatial-temporal semantic coordination module aiming to effectively execute SCD by fusing detailed information with contextual features and harnessing synergies between binary CD and semantic segmentation. Additionally, we introduce a pseudo-label growth algorithm to mitigate the substantial loss of sample category information. The experimental results on two widely used SCD datasets demonstrate that SIC-Net outperforms other methods across various evaluation metrics, achieving SeK of 23.96% and 61.29%, respectively. These findings not only validate the effectiveness of the SIC-Net but also provide new ideas and directions for future research in the field of remote sensing, particularly in SCD.

Abstract Background To evaluate retinal structural and functional changes after silicone oil (SO) removal in eyes with macula-off rhegmatogenous retinal detachment (RRD). Methods Best-corrected visual acuity (BCVA) testing, microperimetry, and optical coherence tomography angiography were performed in 48 eyes with macula-off RRD before and 3 months after SO removal. The values of healthy contralateral eyes were used as control data. Correlations between retinal vessel density (VD), retinal nerve fiber layer thickness (RNFLT), the interval between retinal detachment and surgery, the duration of SO tamponade, the follow-up time after SO removal, and visual function were analyzed. Results Significant increases in 2˚ fixation rate (FR), 4˚ FR, 2˚ mean retinal sensitivity (MRS), 6˚ MRS, parafoveal superficial capillary plexus VD and RNFLT were observed after SO removal (all P 0.05). Conclusion Retinal structure and function improved to different degrees after SO removal. Fixation stability and retinal sensitivity increased more than BCVA postoperatively. Retinal sensitivity, which was affected by the interval between retinal detachment and surgery and the duration of SO tamponade, gradually recovered after SO removal.

Human health and the human microbiome are inevitably intertwined, increasing their relevance in clinical research. However, the collection, transportation and storage of faecal samples may introduce bias due to methodological differences, especially since postal shipping is a common practise in large-scale clinical cohort studies. Using four different Omics layer, we determined the structural (16S rRNA sequencing, cytometric microbiota profiling) and functional integrity (SCFAs, global metabolome) of the microbiota in relation to different easy-to-handle conditions. These conditions were storage at −20 °C, −20 °C as glycerol stock, 4 °C and room temperature with and without oxygen exposure for a maximum of one week. Storage time affected the microbiota on all Omics levels. However, the magnitude was donor-dependent, highlighting the need for purpose-optimized sample collection in clinical multi-donor studies. The effects of oxygen exposure were negligible for all analyses. At ambient temperature, SCFA and compositional profiles were stable for 24 h and 48 h, respectively, while at 4 °C, SCFA profiles were maintained for 48 h. The global metabolome was highly susceptible, already changing at 24 h in non-frozen conditions. Thus, faecal microbiota was best preserved on all levels when transported as a native sample frozen within 24 h, leading to the least biased outcomes in the analysis. We conclude that the immediate freezing of native stool samples for transportation to the lab is best suited for planned multi-Omics analyses that include metabolomics to extend standard sequencing approaches.

Background and Objectives: Fine particulate matter, PM2.5, is becoming a major threat to human health, particularly in terms of respiratory diseases. Pyroptosis is a recently discovered and distinct form of cell death, characterized by pore formation in the cell membrane and secretions of proinflammatory cytokines. There has been little research on the effect of PM2.5 on pyroptosis, especially in airway epithelium. We investigated whether PM2.5-related oxidative stress induces pyroptosis in bronchial epithelial cells and defined the underlying mechanisms. Materials and Methods: After exposure of a BEAS-2B cell line to PM2.5 concentration of 20 µg/mL, reactive oxygen species (ROS) levels, parameters related to pyroptosis, and NF-κB signaling were measured by Western blotting, immunofluorescence, and ELISA (Enzyme-linked immunosorbent assay). Results: PM2.5 induced pyroptotic cell death, accompanied by LDH (Lactate dehydrogenase) release and increased uptake of propidium iodide in a dose-dependent manner. PM2.5 activated the NLRP3-casp1-gasdermin D pathway, with resulting secretions of the proinflammatory cytokines IL-1β and IL-18. The pyroptosis activated by PM2.5 was alleviated significantly by NLRP3 inhibitor. In PM2.5-exposed BEAS-2B cells, levels of intracellular ROS and NF-κB p65 increased. ROS scavenger inhibited the expression of the NLRP3 inflammasome, and the NF-κB inhibitor attenuated pyroptotic cell death triggered by PM2.5 exposure, indicating that the ROS/NF-κB pathway is involved in PM2.5-induced pyroptosis. Conclusions: These findings show that PM2.5 exposure can cause cell injury by NLRP3-inflammasome-mediated pyroptosis by upregulating the ROS/NF-κB pathway in airway epithelium.

Coordination of uranium chemistry by multiligands has attracted attention as a way to obtain unique molecular structures and their possible use in environmental and nuclear waste disposal. In this work, mass spectrum with positive ions ［UO2(N3)n］+(n=0-4) as the dominant peaks generated by laser evaporation of uranium target with N2O as the carrier gas is reported. The associated quantitative calculations are carried out to explore structures and bonding interactions. The calculations show that N3 is bound to UO+2 as a radical and reveal that the bonding interaction in the whole system is found only between the first N3 ligand and uranium atom. Further localized molecular orbital analyses indicate the presence of multi-centered orbital delocalization in the system.

Abstract Biochar has been considered an effective approach as soil amendment for decreasing incidences of disease and regulating microbial populations in continuous-cropping soil. Although researches have extensively focused on changes of soil microbes and unbalance of nutrition in continuous-cropping soil, the relationship between soil properties and pathogens by biochar application remains poorly understood. In this study, we applied ITS ribosomal RNA gene profiling to analyze tobacco root microbiota of biochar and non-biochar treatment in a 3-year continuous-cropping tobacco field, comparing firstly planting tobacco as control. We found that biochar application decreased the relative abundance of the soil fungal pathogens (Ceratobasidium and Monosporascus), which are the prime pathogens of tobacco root rot in continuous-cropping soil. Using RDA, co-occurrence and PLS-PM approaches, we provided evidence that there was a negative correlation between fungal genera (especially for Ceratobasidium and Monosporascus) and soil polyphenol oxidase (PPO) activity (R2 incidence rate = − 0.930, R2 disease index = − 0.905, both p

Pyroptosis represents a type of cell death mechanism notable for its cell membrane disruption and the subsequent release of proinflammatory cytokines. The Nod-like receptor family pyrin domain containing inflammasome 3 (NLRP3) plays a critical role in the pyroptosis mechanism associated with various diseases resulting from particulate matter (PM) exposure. Tert-butylhydroquinone (tBHQ) is a synthetic antioxidant commonly used in a variety of foods and products. The aim of this study is to examine the potential of tBHQ as a therapeutic agent for managing sinonasal diseases induced by PM exposure. The occurrence of NLRP3 inflammasome-dependent pyroptosis in RPMI 2650 cells treated with PM < 4 µm in size was confirmed using Western blot analysis and enzyme-linked immunosorbent assay results for the pyroptosis metabolites IL-1β and IL-18. In addition, the inhibitory effect of tBHQ on PM-induced pyroptosis was confirmed using Western blot and immunofluorescence techniques. The inhibition of tBHQ-mediated pyroptosis was abolished upon nuclear factor erythroid 2-related factor 2 (Nrf2) knockdown, indicating its involvement in the antioxidant mechanism. tBHQ showed potential as a therapeutic agent for sinonasal diseases induced by PM because NLRP3 inflammasome activation was effectively suppressed via the Nrf2 pathway.

Objective To investigate whether platelet-rich plasma (PRP) can promote osteoblasts differentiation in diabetic mellitus(DM) adipose-derived stem cells(ADSCs) by M2 polarization of DM bone marrow derived macrophages(BMDMs). Methods PRP was extracted from the heart of rats according to the instructions of the kit. BMDMs and ADSCs of DM rats were isolated and cultured, then identified by flow cytometry. BMDMs were divided into control group (PRP 0%) and PRP group (1%, 5%, 10%); ADSCs were divided into: control group (control, ADSCs, 2×105/mL, + same volume as PRP group without PRP plasma), ADSCs+BMDMs group (BMDMs group, 2×105/mL, + plasma without PRP), ADSCs+10% PRP group (PRP group, medium with 10% PRP), and ADSCs+BMDMs+10% PRP group (BMDMs+PRP). Transwell was used to count ADSCs by polarized BMDMs in invasion experiment. The polarization of BMDMs and osteogenic differentiation of ADSCs were determined by RT-qPCR and Western blot. The expression of osteogenic protein Osterix was detected by immunofluorescence. The bone defect model of DM rats was constructed with 3 mm×5 mm at femoral epiphysis, and the rats were divided into: control group (DM rats), ADSCs+ clodronate liposomes (CLP)group (ADSCs group), ADSCs+BMDMs group (BMDMs group without CLP injection), ADSCs+CLP+PRP group (PRP group), ADSCs+PRP group (BMDMs+PRP group without CLP injection). CLP(5 mL/kg) was injected into tail vein of rat 48 hours before operation followed by injection twice a week after surgery for 8 weeks. ADSCs (1×107/mL) and alginate gel mixture were injected into the bone defect, and PRP was treated once every 2 weeks with 500 μL PRP for 4 times. Micro-CT was used to reconstruct and quantitatively analyze the bone defect repair in DM rats. Results The purity of ADSCs positive markers Sca-1 and CD29 were 85.4% and 99.9% respectively and and the purity of BMDMs positive markers CD11b and F4/80 were 94% and 90%. Compared with the control group, BMDMs polarized into M2-type dependent on PRP concentration (P＜0.05). Compared with PRP group, the cell counting of ADSCs migrated in BMDMs+PRP group was significantly increased(P＜0.05). Compared with other groups, the osteogenic differentiation activity and the expressions of osteogenic proteins and genes were significantly increased in ADSCs with BMDMs+PRP treatment(P＜0.05). Bone volume/ tissue volume(BV/TV),trabecular number(Tb.N)and trabecular thickness(Tb.Th)were significantly increased in the BMDMs+PRP group compared with the other groups (P＜0.001). Conclusions PRP promotes ADSCs migration and osteogenic differentiation by stimulating the polarization of BMDMs towards M2 in vitro. BMDMs+PRP can significantly promote the differentiation of ADSCs into osteoblasts and accelerate bone formation in vivo, thus improving the poor healing of diabetic bone defects of rats.

Chronic rhinosinusitis (CRS) is a relatively common inflammatory disease of the nasal and paranasal sinus mucosa. Several epidemiological studies have established an association between particulate matter (PM) and CRS. Based on those data, PM has emerged as an important environmental factor in the development of CRS. Recent research has investigated the mechanisms and treatment options for CRS caused by PM through cellular experimentation. Therefore, the authors would like to explain the definition of PM, present research investigating the relationship between PM and CRS, and summarize the involved mechanisms reported to date.

In this study, the amylose contents in 24 commercial foxtail millet samples from major production provinces in China were determined by four different methods, namely, the improved national standard method (GB/T 15683-2008), the spectrophotometric method (NY/T 2639-2014) recommended by China’s agricultural authority, the dual-wavelength method, and the concanavalin A (Con A) method. The results of these methods were in the following decreasing order: improved GB/T 15683-2008 ≈ dual-wavelength > NY/T 2639-2014 > Con A. The correlation coefficient between the results of improved GB/T 15683-2008 and those of NY/T 2639-2014 was highest, 0.752 (P = 0), the former being about 1.41 times higher than the latter, which could allow simple prediction of the amylose content of foxtail millet. In conclusion, the improved GB/T 15683-2008 method, the dual-wavelength method and the NY/T 2639-2014 (× 1.41) method can be used to accurately determine the content of amylose in foxtail millet while greatly reducing sample testing time.

This paper presents a novel model for calculating the dynamic stress of a beam structure in a vibrating screen. The proposed model employs a dynamic screen box model as a foundation and utilizes the flow method to establish the mass of the screening materials within the screen box. By considering the motion characteristics of beam structures, the dynamic model for the beam structure of a vibrating screen is derived. The results indicate that the dynamic model can reveal the internal relationship between the dynamic characteristics of beam structures and the structural and material parameters. Finally, a validation example is presented to demonstrate the effectiveness of the proposed dynamic model. The presented model not only contributes to the understanding of the dynamic behavior of beam structures in vibrating screens but also provides a reliable and accurate method for predicting the dynamic stress in beam structures. The proposed model can be applied to optimize the design and performance of vibrating screens, which have wide applications in mining, metallurgy, and other industries. Further research can be conducted to explore the potential of the proposed model for other types of structures subjected to dynamic loads.