UFObow: A single-wavelength excitable Brainbow for simultaneous multicolor ex-vivo and in-vivo imaging of mammalian cells.

Brainbow is a genetic cell-labeling technique that allows random colorization of multiple cells and real-time visualization of cell fate within a tissue, providing valuable insights into understanding complex biological processes. However, fluorescent proteins (FPs) in Brainbow have distinct excitation spectra with peak difference greater than 35 nm, which requires sequential imaging under multiple excitations and thus leads to long acquisition times. In addition, they are not easily used together with other fluorophores due to severe spectral bleed-through. Here, we report the development of a single-wavelength excitable Brainbow, UFObow, incorporating three newly developed blue-excitable FPs. We have demonstrated that UFObow enables not only tracking the growth dynamics of tumor cells in vivo but also mapping spatial distribution of immune cells within a sub-cubic centimeter tissue, revealing cell heterogeneity. This provides a powerful means to explore complex biology in a simultaneous imaging manner at a single-cell resolution in organs or in vivo.

Groundwater hydrochemical signatures, nitrate sources, and potential health risks in a typical karst catchment of North China using hydrochemistry and multiple stable isotopes.

Nitrate pollution in aquatic ecosystems has received growing concern, particularly in fragile karst basins. In this study, hydrochemical compositions, multiple stable isotopes (δ2H-H2O, δ18Ο-Η2Ο, δ15Ν-ΝΟ3-, and δ18Ο-ΝΟ3-), and Bayesian stable isotope mixing model (MixSIAR) were applied to elucidate nitrate pollution sources in groundwater of the Yangzhuang Basin. The Durov diagram identified the dominant groundwater chemical face as Ca-HCO3 type. The NO3- concentration ranged from 10.89 to 90.45 mg/L (average 47.34 mg/L), showing an increasing trend from the upstream forest and grassland to the downstream agricultural dominant area. It is worth noting that 47.2% of groundwater samples exceeded the NO3- threshold value of 50 mg/L for drinking water recommended by the World Health Organization. The relationship between NO3-/Cl- and Cl- ratios suggested that most groundwater samples were located in nitrate mixed endmember from agricultural input, soil organic nitrogen, and manure & sewage. The Self-Organizing Map (SOM) and Pearson correlations analysis further indicated that the application of calcium fertilizer, sodium fertilizer, and livestock and poultry excrement in farmland elevated NO3- level in groundwater. The output results of the MixSIAR model showed that the primary sources of NO3- in groundwater were soil organic nitrogen (55.3%), followed by chemical fertilizers (28.5%), sewage & manure (12.7%), and atmospheric deposition (3.4%). Microbial nitrification was a dominant nitrogen conversion pathway elevating NO3- levels in groundwater, while the denitrification can be neglectable across the study area. The human health risk assessment (HHRA) model identified that about 88.9%, 77.8%, 72.2%, and 50.0% of groundwater samples posing nitrate's non-carcinogenic health hazards (HQ > 1) through oral intake for infants, children, females, and males, respectively. The findings of this study can offer useful biogeochemical information on nitrogen pollution in karst groundwater to support sustainable groundwater management in similar human-affected karst regions.

Hydrogeochemical characterization, quality assessment, and potential nitrate health risk of shallow groundwater in Dongwen River Basin, North China.

Assessing groundwater geochemical formation processes and pollution circumstances is significant for sustainable watershed management. In the present study, 58 shallow groundwater samples were taken from the Dongwen River Basin (DRB) to comprehensively assess the hydrochemical sources, groundwater quality status, and potential risks of NO3- to human health. Based on the Box and Whisker plot, the cation's concentration followed the order of Ca2+ > Mg2+ > Na+ > K+, while anions' mean levels were HCO3- > SO42- > NO3- > Cl-. The NO3- level in groundwater samples fluctuated between 4.2 and 301.3 mg/L, with 67.2% of samples beyond the World Health Organization (WHO) criteria (50 mg/L) for drinking. The Piper diagram indicated the hydrochemical type of groundwater and surface water were characterized as Ca·Mg-HCO3 type. Combining ionic ratio analysis with principal component analysis (PCA) results, agricultural activities contributed a significant effect on groundwater NO3-, with soil nitrogen input and manure/sewage inputs also potential sources. However, geogenic processes (e.g., carbonates and evaporite dissolution/precipitation) controlled other ion compositions in the study area. The groundwater samples with higher NO3- values were mainly found in river valley regions with intense anthropogenic activities. The entropy weight water quality index (EWQI) model identified that the groundwater quality rank ranged from excellent (70.7%) and good (25.9%) to medium (3.4%). However, the hazard quotient (HQ) used in the human health risk assessment (HHRA) model showed that above 91.38% of groundwater samples have a NO3- non-carcinogenic health risk for infants, 84.48% for children, 82.76% for females, and 72.41% for males. The findings of this study could provide a scientific basis for the rational development and usage of groundwater resources as well as for the preservation of the inhabitants' health in DRB.

Identifying the hydrochemical features, driving factors, and associated human health risks of high-fluoride groundwater in a typical Yellow River floodplain, North China.

Fluoride enrichment (> 1.5 mg/L) in groundwater has become a global threat, particularly given the hazards to human health. This study collected 58 unconfined groundwater samples from Fengpei Plain in June 2022 for hydrochemical and stable isotope analyses combined with multiple methods to explore sources, influencing factors, and potential health hazards of groundwater F-. The results showed that groundwater F- concentration ranged from 0.08 to 8.14 mg/L, with an average of 1.91 mg/L; over 41.4% of them exceeded the acceptable level of 1.5 mg/L prescribed by the World Health Organization (WHO). The dominant hydrochemical facies changed from Ca·Mg-HCO3 and Ca·Mg-SO4·Cl type in low-F- groundwater to Na-HCO3 and Na-SO4·Cl water types in high-F- groundwater. The Self-Organizing Map (SOM) and ionic correlation analysis indicated that F- is positively correlated to pH, EC, Na+, K+, SO42-, and TDS, but negatively to Ca2+ and δ18O. Groundwater F- accumulation was primarily driven by F--bearing minerals dissolution such as fluorite. Simultaneously, the carbonates precipitation, positive cation exchange processes, and salt effect were conducive to groundwater F- enrichment. However, competitive adsorption between OH-/HCO3- and F-, evaporation, and anthropogenic activities only had a weak effect on the F- enrichment in groundwater. The hazard quotient (HQ) assessment results show that 67.2% of groundwater samples pose a non-carcinogenic risk (HQ > 1) for infants, followed by 53.4% for children, 32.8% for females, and 25.9% for males. The Monte Carlo simulation results agreed with those of the deterministic model that minors are more susceptible than adults. These findings are vital to providing insights into the geochemical behavior, driving factors, and drinking water safety of high-F- groundwater worldwide.

Three-dimensional mapping of hepatic lymphatic vessels and transcriptome profiling of lymphatic endothelial cells in healthy and diseased livers.

Rationale: Hepatic lymphatics are essential for liver homeostasis and immune function. However, the 3D structure and spatial distribution of hepatic lymphatic vessels (LVs) need to be confirmed. Moreover, the molecular information of hepatic lymphatic endothelial cells (LyECs) needs to be further studied. The bottleneck is the lack of specific markers or labeling methods for hepatic lymphatic endothelial cells (LyECs) Methods: Here, we proposed a method for the spatiotemporal sequential injection of antibodies (STSI-Ab) to selectively label hepatic LyECs in vivo. In addition, we also developed an efficient hepatic LyEC sorting method and performed deep transcriptome sequencing on hepatic LyECs. Results: The STSI-Ab method achieved selective labeling of the mouse hepatic lymphatic network. Three-dimensional fluorescence imaging results of the STSI-Ab mouse liver lobe clearly showed that hepatic LVs entangled with the portal vein but were not present in the central vein. The imaging data inspired a novel hepatic lobule structure model with an added set of LVs in the portal area. Furthermore, deep transcriptome sequencing of isolated hepatic LyECs and Masson's trichrome staining results suggested that hepatic LyECs might be an important source of collagen fibers deposited in the portal area during the process of liver fibrosis and bile duct ligation (BDL). Conclusions: We proposed an STSI-Ab method for selectively labeling hepatic LVs, distinguishing the hepatic LVs from other vessels, and mapping their 3D structure. This study opens an avenue for understanding hepatic lymphatic structure and it will be very beneficial to the study of hepatic LyEC functions.

Surface molecularly imprinted polymers with synthetic dummy template for simultaneously selective recognition of nine phthalate esters.

Dummy template molecularly imprinted polymers (DMIPs) on silica gel particles for simultaneously selective recognition of nine phthalate esters have been prepared. A novel dummy template molecule with similar structural skeleton to the phthalate ester, diethyl N,N'-phthaloyl-bis(11-aminoundecanoate), has been designed and synthesized. The DMIP films were grafted on the surface of silica gel particles by a sol-gel process with 3-aminopropyltriethoxysilane (APTES) and tetramethoxysilane (TEOS) as functional monomer and cross-linker, respectively, and the obtained sorbents have been characterized by FTIR with diffuse reflectance accessory. The maximum static adsorption capacities of the DMIPs and NIP sorbents for the nine phthalate esters were 281 and 132mg/g respectively, and the results of dynamic adsorption showed that the adsorption equilibrium could be achieved about 5min for the DMIPs sorbents. The imprint factors of the sorbents ranged from 1.8 to 3.0 for eight of the phthalate esters except for Diamyl phthalate, which indicated that the DMIPs sorbents have high selectivity. The competitive experiments of the nine phthalates with some of their analogues on the sorbents illustrated that the DMIPs sorbents have high specificity for the phthalates. A GC-MS method for determination of the phthalate residues in fruit juice have been developed with the DMIPs as sorbents for the solid phase extraction (SPE) in the sample pretreatment procedures. The spiking recoveries of the phthalates were in the range of 72-100.2% with relative standard deviations lower than 10.2%. The results indicated that the obtained sorbents could specifically recognize the phthalates from complex matrices, which provide a new train of thought for preparing the DMIPs sorbents.

Determination of emulsion explosives with Span-80 as emulsifier by gas chromatography-mass spectrometry.

A novel approach for identification and determination of emulsion explosives with Span-80 (sorbitol mono-oleate) as the emulsifier and their postblast residues by gas chromatography-mass spectrometry (GC-MS) has been developed. 24 kinds of emulsion explosives collected have been processed by transesterification reaction with metholic KOH solution and the emulsifier has turned into methyl esters of fatty acids. From the peak area ratios of their methyl esters, most of these emulsion explosives can be differentiated. In order to detect the postblast residues of emulsion explosives, the sorbitols in the emulsifier Span-80 obtained after transesterification reaction have been further derivatized by silylation reaction with N,O-bis-(trimethylsilyl)trifluoroacetamide (BSTFA) containing 1% trimethylchlorosilane (TMCS) as the derivatizing reagent. The derivatization conditions were optimized and the derivatives were determined by GC-MS. The results showed that the silylation derivatives of sorbitol and it isomers, combined with hydrocarbon compounds and methyl esters of fatty acids, were the characteristic components for identification of the emulsion explosives. The established approach was applied to analyze the postblast residues of emulsion explosives. It has been found that the method was sensitive and specific, especially when detecting the derivatives of sorbitol and its isomers by GC-MS in selecting ion mode. The information of the characteristic components can help probe the origin of the emulsion explosives and providing scientific evidences and clues for solving the crimes of the emulsion explosive explosion.