TLR3 Regulated Poly I:C-Induced Neutrophil Extracellular Traps and Acute Lung Injury Partly Through p38 MAP Kinase

Acute lung injury (ALI) is the leading cause of morbidity and mortality in critically ill patients. Neutrophil extracellular traps (NETs) have been well documented in the ALI model of bacterial infection. In the present study, we demonstrated that poly I:C could induce pulmonary NETs. Upon poly I:C intratracheal inoculation, neutrophil infiltration in the bronchoalveolar lavage fluid (BALF) was significantly increased. Furthermore, the inflammatory cytokines IL-1β, IL-6, and TNF-α in the lung were also significantly elevated. Neutrophil depletion abolished NETs and decreased both neutrophil infiltration and IL-1β in the lung. As expected, DNase I, an inhibitor of MPO and NADPH, decreased pulmonary inflammation and NETs. Blocking of the poly I:C receptor TLR3 reduced lung inflammation and NETs. The MAPK kinase inhibitor p38 diminished the formation of NETs and restored the expression of the tight junction protein claudin-5 in the mouse lung when challenged with poly I:C. In summary, poly I:C induced the formation of pulmonary NETs and ALI, which may be associated with the activation of p38 MAPK and the decreased expression of claudin-5

Generation of integration-free neural progenitor cells from cells in human urine

Human neural stem cells hold great promise for research and therapy in neural disease. We describe the generation of integration-free and expandable human neural progenitor cells (NPCs). We combined an episomal system to deliver reprogramming factors with a chemically defined culture medium to reprogram epithelial-like cells from human urine into NPCs (hUiNPCs). These transgene-free hUiNPCs can self-renew and can differentiate into multiple functional neuronal subtypes and glial cells in vitro. Although functional in vivo analysis is still needed, we report that the cells survive and differentiate upon transplant into newborn rat brain.postprin

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Flood Risk Assessment of Subway Systems in Metropolitan Areas under Land Subsidence Scenario: A Case Study of Beijing

Flooding is one of the most destructive natural events that severely damage the ground and inundate underground infrastructure. Subway systems in metropolitan areas are susceptible to flooding, which may be exacerbated when land subsidence occurs. However, previous studies have focused on flood risk evaluation on regional/watershed-scales and land subsidence monitoring in plains, instead of on subway flood risk evaluation and how land subsidence aggravates the flood risk in subway systems. Using the proposed risk indicators and field survey data, we present a method assessing the flood risk of metropolitan subway systems under a subsidence condition based on the fuzzy analytic hierarchy process (FAHP) combined with a geographic information system (GIS). We use the regional risk level within the 500 m buffer zone of the subway line to depict the flood risk of the subway system. The proposed method was used to evaluate the flood risk of the Beijing subway system. The results show that the flood risks of the Beijing subway show a ring-like distribution pattern—risk levels decreasing from the central urban area to the suburbs. Very high and high risks are mainly located within third and fourth ring roads, accounting for 63.58% (29.40 km2) and 63.83% (81.19 km2) of the total area. Land subsidence exacerbated the Beijing subway system’s flood risk level—the moderate to very high risk increased by 46.88 km2 (16.33%), indicating that land subsidence is an essential factor affecting the flood risk level of subway systems. In addition to enhancing flood warnings, future subway flooding could be reduced by elevating the height of the stations’ exit (entrance) and installing water stop plates and watertight doors. This study is of great significance for flood warning and prevention in the Beijing subway system; it provides a theoretical basis for flood risk evaluation in other metropolitan areas

Transcriptional profiling of innate immune responses in sheep PBMCs induced by Haemonchus contortus soluble extracts

Abstract Background Pattern recognition receptors (PRRs) can recognize pathogen-associated molecular patterns and activate downstream signalling pathways, resulting in modulation of host immunity against pathogens. Here, we investigated whether PRR-mediated recognition is involved in host immune responses to the blood-feeding nematode Haemonchus contortus. Methods During blood-feeding, H. contortus secretes immune-modulating antigens into host blood. Therefore, we stimulated sheep peripheral blood mononuclear cells (PBMCs) with H. contortus soluble extract (HcAg) and performed transcriptional profiling. Results HcAg upregulated two genetically linked CLRs (CLEC2L and KLRG2), two NLRs attenuating inflammation (NLRP12 and NLRC3) and one G protein-coupled receptor with potent anti-inflammatory effects (HCAR2). Furthermore, several Th2-related transcription factors (ATF3, IRF4, BCL3 and NFATC) were also upregulated, which may confer anti-inflammatory type 2 immune responses to HcAg. Conclusions Together, our preliminary studies provide new insights into how the host innate immune system controls type 2 immunity to H. contortus. Further work will be needed to identify H. contortus products recognized by the host innate immune system and determine the Th2 polarization ability of these putative PRR ligands

Flood Risk Assessment Based on Fuzzy Synthetic Evaluation Method in the Beijing-Tianjin-Hebei Metropolitan Area, China

Flooding is one of the most devastating natural events and leads to enormous and recurring loss of life, properties, and resources around the globe. With climate change and accelerating urbanization, flood disasters in China have increasingly affected the sustainable development of metropolitan areas. Risk assessment is an essential step in flood management and disaster mitigation, which provide a quantitative measure of flood risk. However, the difficulty of flood risk zoning is dealing with the uncertainty of the evaluation process and the complicated non-linear relationship between indicators and risk levels. To address this issue, a fuzzy synthetic evaluation (FSE) method based on combined weight (CW) was utilized in this paper to generate flood risk maps at a grid-scale (1 × 1 km). For the case study in the Beijing-Tianjin-Hebei metropolitan area (BTH) in China, fourteen indicators were selected to construct the flood risk assessment model based on the FSE approach integrated with ArcGIS. The research demonstrates that moderate, high, and very high risk zones are distributed in the southeast fluvial plain of the BTH area, accounting for 31.36% of the total land area. Meanwhile, low and very-low risk zones occupy 68.64% of the total land area, and are primarily located in the high plateau and mountain regions in the northwest. We analyzed the risk level of each county and proposed risk mitigation measures based on field investigations. The verified risk assessment results were spatially consistent with the historical flood disaster records and loss positions, indicating the accuracy and reliability of the risk assessment map using the FSE approach. Compared with the IPCC (Intergovernmental Panel on Climate Change) TAR (Third Assessment Report) and AR5 (Fifth Assessment Report) methods, FSE has significant advantages in handling uncertainty, complexity, and the non-linear relationship between indices and risk grades. This study provides a novel quantitative method for flood risk assessment in metropolitan areas and practical implications for urban flood management