Kindlin-2 promotes rear focal adhesion disassembly and directional persistence during cell migration.

Cell migration involves front-to-rear asymmetric focal adhesion (FA) dynamics, which facilitates trailing edge detachment and directional persistence. Here, we show that kindlin-2 is crucial for FA sliding and disassembly in migrating cells. Loss of kindlin-2 markedly reduced FA number and selectively impaired rear FA sliding and disassembly, resulting in defective rear retraction and reduced directional persistence during cell migration. Kindlin-2-deficient cells failed to develop serum-induced actomyosin-dependent tension at FAs. At the molecular level, kindlin-2 directly interacted with myosin light chain kinase (MYLK, hereafter referred to as MLCK), which was enhanced in response to serum stimulation. Serum deprivation inhibited rear FA disassembly, which was released in response to serum stimulation. Overexpression of the MLCK-binding kindlin-2 F0F1 fragment (amino acid residues 1-167), which inhibits the interaction of endogenous kindlin-2 with MLCK, phenocopied kindlin-2 deficiency-induced migration defects. Inhibition of MLCK, like loss of kindlin-2, also impaired trailing-edge detachment, rear FA disassembly and directional persistence. These results suggest a role of kindlin-2 in promoting actomyosin contractility at FAs, leading to increased rear FA sliding and disassembly, and directional persistence during cell migration.

Clinical and Genetic Characteristics of a Cohort with Distal Vaginal Atresia.

Distal vaginal atresia is a rare abnormality of female reproductive tract in which the vagina is closed or absent. The distal vagina may be replaced by fibrous tissue and the condition is often not diagnosed until a girl fails to begin having periods at puberty. Although it is a congenital disorder, potential genetic causes of distal vaginal atresia are still unknown. We recruited a cohort of 39 patients with distal vaginal atresia and analyzed their phenotypic and genetic features. In addition to the complaint of distal vaginal atresia, approximately 17.9% (7/39) of the patients had other Müllerian anomalies, and 17.9% (7/39) of the patients had other structural abnormalities, including renal-tract, skeletal and cardiac anomalies. Using genome sequencing, we identified two fragment duplications on 17q12 encompassing HNF1B and LHX1, two dosage-sensitive genes with candidate pathogenic variants, in two unrelated patients. A large fragment of uniparental disomy was detected in another patient, affecting genes involved in cell morphogenesis and connective tissue development. Additionally, we reported two variants on TBX3 and AXL, leading to distal vaginal atresia in mutated mouse model, in our clinical subjects for the first time. Essential biological functions of these detected genes with pathogenic variants included regulating reproductive development and cell fate and patterning during embryogenesis. We displayed the comprehensive clinical and genetic characteristic of a cohort with distal vaginal atresia and they were highly heterogeneous both phenotypically and genetically. The duplication of 17q12 in our cohort could help to expand its phenotypic spectrum and potential contribution to the distal vaginal atresia. Our findings of pathogenic genetic variants and associated phenotypes in our cohort could provide evidence and new insight for further research attempting to reveal genetic causes of distal vaginal atresia.

Characterization of gut microbiota associated with clinical parameters in intrahepatic cholestasis of pregnancy.

BACKGROUND: Intrahepatic cholestasis of pregnancy (ICP) is a liver disorder that specifically occurs in pregnancy. Elevated levels of liver transaminases aspartate aminotransferase, alanine aminotransferase and serum bilirubin levels are common biochemical characteristics in ICP. The disorder is associated with an increased risk of premature delivery and stillbirth. The characterization of the potential microbiota in ICP could go a long way in the prevention and treatment of this pregnancy disease. METHODS: A total of 58 patients were recruited for our study: 27 ICP patients and 31 healthy pregnant subjects with no ICP. The V3 and V4 regions of the 16S rDNA collected from fecal samples of both diseased and control groups were amplified. 16S rRNA gene amplicon sequencing was then performed on gut microbiota. Sequencing data were analyzed and the correlation between components of microbiota and patient ICP status was found. Related metabolic pathways, relative abundance and significantly different operational taxonomic units (OTUs) between ICP and controls were also identified. RESULTS: Elevated levels of total bile acid, ALT, AST, Dbil and Tbil were recorded or observed in ICP subjects as compared to the control. Gut microbiota in pregnant women was dominated by four major phyla and 27 core genera. PCoA analysis results indicated that there was no significant clustering in Bray-Curtis distance matrices. Our results showed that there was a correlation between specific OTUs and measured clinical parameters of pregnant women. Comparison at the different taxonomy levels revealed high levels of abundance of Blautia and Citrobacter in ICP patients. At the family level, Enterobacteriaceae and Leuconostocaceae were higher in ICP patients. 638 KEGG Orthologs and 138 pathways significantly differed in the two groups. PLS-DA model with VIP plots indicated a total of eight genera and seven species were key taxa in ICP and control groups. CONCLUSIONS: Our research indicated that although there was no significant clustering by PCoA analysis, patients with ICP have increased rare bacteria at different phylogenetic levels. Our results also illustrated that all 638 KEGG Orthologs and 136 in 138 KEGG pathways were less abundant in ICP patients compared to the controls.

The ratio of H/C is a useful parameter to predict adsorption of the herbicide metolachlor to biochars.

Biochar adsorbent was produced by pyrolyzing traditional Chinese medicinal herb residue at 300, 500 and 750 °C (referred to as biochar-300, biochar-500 and biochar-750). Basic physical and chemical analyses, Fourier transform infrared spectroscopy (FT-IR), and thermodynamic analyses were performed to elucidate adsorption and properties of biochar. Biochar adsorption capacity of herbicide metolochlor, as measured by batch-type adsorption experiments by Freundlich constant Kf (mg1-n Ln kg-1), followed the order: biochar-750 > biochar-300 > biochar-500. Thermodynamic analysis suggested that adsorption of metolachlor on biochar was a spontaneous process. The adsorption isotherm for the biochar produced at the highest pyrolysis temperature was characteristic for adsorption process driven by a high surface area of biochar (85.30 m2 g-1), while the adsorption process for the biochar produced at the lowest temperature was controlled by its higher content of organic matter (39.06%) and abundant functional groups. The FT-IR spectra also showed that the biochar prepared at the lowest temperature had the highest number of surface groups. In general, pore-filling induced by the large surface area of the biochar was the dominant adsorption mechanism. When the H/C value was >0.5, the adsorption mechanism of biochar was dominated by surface chemical bond, while pore-filling played a major role when the H/C value was <0.5.

Kindlin-2 Association with Rho GDP-Dissociation Inhibitor α Suppresses Rac1 Activation and Podocyte Injury.

Alteration of podocyte behavior is critically involved in the development and progression of many forms of human glomerular diseases. The molecular mechanisms that control podocyte behavior, however, are not well understood. Here, we investigated the role of Kindlin-2, a component of cell-matrix adhesions, in podocyte behavior in vivo Ablation of Kindlin-2 in podocytes resulted in alteration of actin cytoskeletal organization, reduction of the levels of slit diaphragm proteins, effacement of podocyte foot processes, and ultimately massive proteinuria and death due to kidney failure. Through proteomic analyses and in vitro coimmunoprecipitation experiments, we identified Rho GDP-dissociation inhibitor α (RhoGDIα) as a Kindlin-2-associated protein. Loss of Kindlin-2 in podocytes significantly reduced the expression of RhoGDIα and resulted in the dissociation of Rac1 from RhoGDIα, leading to Rac1 hyperactivation and increased motility of podocytes. Inhibition of Rac1 activation effectively suppressed podocyte motility and alleviated the podocyte defects and proteinuria induced by the loss of Kindlin-2 in vivo Our results identify a novel Kindlin-2-RhoGDIα-Rac1 signaling axis that is critical for regulation of podocyte structure and function in vivo and provide evidence that it may serve as a useful target for therapeutic control of podocyte injury and associated glomerular diseases.

Heritable CRISPR/Cas9-mediated targeted integration in Xenopus tropicalis.

Xenopus tropicalis is an emerging vertebrate genetic model. A gene knock-in method has not yet been reported in this species. Here, we report that heritable targeted integration can be achieved in this diploid frog using a concurrent cleavage strategy mediated by the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 9 (CRISPR/Cas9) system. The key point of the strategy is the addition of a Cas9/guide RNA cleavage site in the donor vector, allowing simultaneous cutting of the chromosomal target site and circular donor DNA in vivo. For the 3 distinct loci tested, all showed efficient targeted integration that was verified by both germ-line transmission and Southern blot analyses. By designing the target sites in introns, we were able to get precise editing of the tyrosinase coding sequence and green fluorescent protein expression from endogenous n-tubulin promoter and enhancers. We were unable to detect off-target effects with the T7 endonuclease I assay. Precise editing of protein coding sequences in X. tropicalis expands the utility of this diploid frog, such as for establishing models to study human inherited diseases.

Integrated miRNA and mRNA expression profiling in fetal hippocampus with Down syndrome.

BACKGROUNDS: Down syndrome (DS), caused by triplication of human chromosome 21, is the most common aneuploidies. The main characteristic of DS patients is intellectual disability. MicroRNAs (miRNAs) play important regulatory roles in various biological processes, such as embryonic development, cell differentiation, proliferation and apoptosis. Several miRNAs have shown association with DS. However, the role of miRNAs in DS patients has not been well elaborated. METHODS: In this research, total RNA extracted from fetal hippocampal tissues was used to analyze miRNA and mRNA expression via Affymetrix miRNA 4.0 and PrimeView Human Gene Expression Array, respectively. Then miRNA and gene expression profiles were integrated by correlation analysis to identify dysregulated miRNAs with their predicted target mRNAs. Microarray data were further validated by real-time PCR. Regulation of zeste homolog 2 (EZH2) expression by hsa-miR-138 was determined by luciferase reporter assay. RESULTS: We analyzed microRNA expression profile in hippocampal tissues from DS fetal using miRNA microarray. Further with the use of mRNA microarray data, we integrate miRNA expression and mRNA expression in hippocampus of trisomy 21 fetus to elucidate the mechanism that underlying DS abnormalities. We characterized the repertoire of specific miRNAs involved in hippocampus in trisomy 21 patients, highlighting hsa-miR-138 and hsa-miR-409, in particular the importance of hsa-miR-138, especially the -5p strand. Furthermore, the expression level of predicted target genes of hsa-miR-138-5p in trisomy 21 fetus, including zeste homolog 2 (EZH2) were further confirmed. In addition, luciferase assay indicated that EZH2 was a direct target of hsa-miR-138 in HEK293T cells. CONCLUSION: The function of hsa-miR-138-5p and its target EZH2 was involved in hippocampus in DS patients. Our findings provide a clue to study the underlying molecular mechanisms in DS patients, and its potential contribution in improving understanding of intellectual disability development in DS patients.

Field-scale fluorescence fingerprinting of biochar-borne dissolved organic carbon.

Biochar continues to receive worldwide enthusiasm as means of augmenting recalcitrant organic carbon in agricultural soils. Realistic biochar amendment rate (typically less than 1 wt%) in the field scale, and subsequent loss by sizing, rain, and other transport events demand reliable methods to quantify the remaining portions of amended biochar. This study employed fluorescence excitation-emission (EEM) spectrophotometry and parallel factor analysis (PARAFAC) to specifically target pyrogenic dissolved organic carbon (DOC) released by amended biochar during the course of a field trial at Bowling Green, KY experimental site. Toluene/methanol (1:6 v/v) extracts of surface (0-15 cm) soils amended with 21.28 t ha(-1) fast pyrolysis biochar afforded PARAFAC fingerprints representing different degrees of aromaticity. Compared to the control without treatments, biochar treatment (with and without poultry manure or chemical fertilizer) increased the relative contribution of PARAFAC fingerprint attributable to labile polyaromatic DOC structures. Poultry manure or chemical fertilizer alone (without biochar) did not influence the amounts of polyaromatic DOC structures. Existence of biochar could be further validated by the changes in %DOC (relative to the total carbon), fixed C content, and UV absorbance (360 nm), whereas FTIR, %O, and sorption of model agrochemical (deisopropylatrazine) did not reflect the presence of biochar in the soil samples. Developed toluene/methanol-based EEM-PARAFAC technique will provide a sensitive, rapid, and cost-competitive method to validate the long-term carbon sequestration by the biochar soil amendment.

Biochar Blended with Alkaline Mineral Can Better Inhibit Lead and Cadmium Uptake and Promote the Growth of Vegetables.

Three successive vegetable pot experiments were conducted to assess the effects on the long-term immobilization of heavy metals in soil and crop yield improvement after the addition of peanut shell biochar and an alkaline mineral to an acidic soil contaminated with lead and cadmium. Compared with the CK treatment, the change rates of biomass in the edible parts of the three types of vegetables treated with B0.3, B1, B3, B9, R0.2 and B1R0.2 were -15.43%~123.30%, 35.10%~269.09%, 40.77%~929.31%, -26.08%~711.99%, 44.14%~1067.12% and 53.09%~1139.06%, respectively. The cadmium contents in the edible parts of the three vegetables treated with these six additives reduced by 2.08%~13.21%, 9.56%~24.78%, 9.96%~35.61%, 41.96%~78.42%, -4.19%~57.07% and 12.43%~65.92%, respectively, while the lead contents in the edible parts reduced by -15.70%~59.47%, 6.55%~70.75%, 3.40%~80.10%, 55.26%~89.79%, 11.05%~70.15% and 50.35%~79.25%, respectively. Due to the increases in soil pH, soil cation-exchange capacity and soil organic carbon content, the accumulation of Cd and Pb in the vegetables was most notably reduced with a high dosage of 9% peanut shell biochar alone, followed by the addition of a low dosage of 1% peanut shell biochar blended with 0.2% alkaline mineral. Therefore, the addition of a low dosage of 1% peanut shell biochar blended with 0.2% alkaline mineral was the best additive in increasing the vegetable biomass, whereas the addition of 9% peanut shell biochar alone was the worst. Evidently, the addition of 0.2% alkaline mineral can significantly reduce the amount of peanut shell needed for passivating heavy metals in soil, while it also achieves the effect of increasing the vegetable yield.

Predictive Machine Learning Model to Assess the Adsorption Efficiency of Biochar-Heavy Metals for Effective Remediation of Soil-Plant Environment.

Biochar is crucial for agricultural output and plays a significant role in effectively eliminating heavy metals (HMs) from the soil, which is essential for maintaining a soil-plant environment. This work aimed to assess machine learning models to analyze the impact of soil parameters on the transformation of HMs in biochar-soil-plant environments, considering the intricate non-linear relationships involved. A total of 211 datasets from pot or field experiments were evaluated. Fourteen factors were taken into account to assess the efficiency and bioavailability of HM-biochar amendment immobilization. Four predictive models, namely linear regression (LR), partial least squares (PLS), support vector regression (SVR), and random forest (RF), were compared to predict the immobilization efficiency of biochar-HM. The findings revealed that the RF model was created using 5-fold cross-validation, which exhibited a more reliable prediction performance. The results indicated that soil features accounted for 79.7% of the absorption of HM by crops, followed by biochar properties at 17.1% and crop properties at 3.2%. The main elements that influenced the result have been determined as the characteristics of the soil (including the presence of different HM species and the amount of clay) and the quantity and attributes of the biochar (such as the temperature at which it was produced by pyrolysis). Furthermore, the RF model was further developed to predict bioaccumulation factors (BAF) and variations in crop uptake (CCU). The R2 values were found to be 0.7338 and 0.6997, respectively. Thus, machine learning (ML) models could be useful in understanding the behavior of HMs in soil-plant ecosystems by employing biochar additions.

Quantitative Soil Characterization for Biochar-Cd Adsorption: Machine Learning Prediction Models for Cd Transformation and Immobilization.

Soil pollution with cadmium (Cd) poses serious health and environmental consequences. The study investigated the incubation of several soil samples and conducted quantitative soil characterization to assess the influence of biochar (BC) on Cd adsorption. The aim was to develop predictive models for Cd concentrations using statistical and modeling approaches dependent on soil characteristics. The potential risk linked to the transformation and immobilization of Cd adsorption by BC in the soil could be conservatively assessed by pH, clay, cation exchange capacity, organic carbon, and electrical conductivity. In this study, Long Short-Term Memory (LSTM), Bidirectional Gated Recurrent Unit (BiGRU), and 5-layer CNN Convolutional Neural Networks (CNNs) were applied for risk assessments to establish a framework for evaluating Cd risk in BC amended soils to predict Cd transformation. In the case of control soils (CK), the BiGRU model showed commendable performance, with an R2 value of 0.85, indicating an approximate 85.37% variance in the actual Cd. The LSTM model, which incorporates sequence data, produced less accurate results (R2=0.84), while the 5-layer CNN model had an R2 value of 0.91, indicating that the CNN model could account for over 91% of the variation in actual Cd levels. In the case of BC-applied soils, the BiGRU model demonstrated a strong correlation between predicted and actual values with R2 (0.93), indicating that the model explained 93.21% of the variance in Cd concentrations. Similarly, the LSTM model showed a notable increase in performance with BC-treated soil data. The R2 value for this model stands at a robust R2 (0.94), reflecting its enhanced ability to predict Cd levels with BC incorporation. Outperforming both recurrent models, the 5-layer CNN model attained the highest precision with an R2 value of 0.95, suggesting that 95.58% of the variance in the actual Cd data can be explained by the CNN model's predictions in BC-amended soils. Consequently, this study suggests developing ecological soil remediation strategies that can effectively manage heavy metal pollution in soils for environmental sustainability.

Protocol for optimized nasal mucosa sample processing to obtain high-quality scRNA-seq and scATAC-seq data.

Examining nasal mucosa samples is crucial for nasal cavity disease research and diagnosis. Simultaneously obtaining high-quality data for single-cell transcriptomics (single-cell RNA sequencing [scRNA-seq]) and epigenomics (single-cell assay for transposase-accessible chromatin using sequencing [scATAC-seq]) of nasal mucosa tissues is challenging. Here, we present a protocol for processing human nasal mucosa samples to obtain data for both scRNA-seq and scATAC-seq. We describe steps for extracting human nasal mucosa tissue, mechanical and enzymatic dissociation, lysis of red blood cells, and a viability assay. We then detail procedures for library preparation and quality control.

A Chinese patient with Rothmund-Thomson syndrome.

INTRODUCTION: Rothmund-Thomson syndrome (RTS) is a rare autosomal recessive disorder that has been reported in all ethnicities, with several identifiable pathogenic variants. There have been reported cases indicating that RTS may lead to low birth weight in fetuses, but specific data on the fetal period are lacking. Genetic testing for RTS II is currently carried out by identifying pathogenic variants in RECQL4. METHODS: In order to determine the cause, we performed whole-genome sequencing (WGS) analysis on the patient and his parents. Variants detected by WGS were confirmed by Sanger sequencing and examined in family members. RESULTS: After analyzing the WGS data, we found a heterozygous nonsense mutation c.2752G>T (p.Glu918Ter) and a novel frameshift insertion mutation c.1547dupC (p.Leu517AlafsTer23) of RECQL4, which is a known pathogenic/disease-causing variant of RTS. Further validation indicated these were compound heterozygous mutations from parents. CONCLUSION: Our study expands the mutational spectrum of the RECQL4 gene and enriches the phenotype spectrum of Chinese RTS patients. Our information can assist the patient's parents in making informed decisions regarding their future pregnancies. This case offers a new perspective for clinicians to consider whether to perform prenatal diagnosis.

Biochar Derived from Urban Green Waste Can Enhance the Removal of Cd from Water and Reduce Soil Cd Bioavailability.

The beneficial utilization of potentially increasing urban green waste (UGW) is critical for sustainable urban development in China. In this study, UGW was pyrolyzed at different temperatures, and the resulting biochar was used to amend Cd-contaminated soils to grow cabbage. Our results showed that the Cd adsorption capacity of UGW-biochar was positively correlated with the surface area, O/C, and (O+N)/C value of biochar. Furthermore, UGW-biochar was incorporated into three Cd-contaminated soils, including one acidic soil and two neutral soils, to assess its impact on the availability of Cd. The most substantial reduction in the concentration of available Cd was observed in the acidic soil, of the three tested soils. In the neutral soils, a more substantial reduction was found in the heavily Cd-contaminated soil compared to the lightly Cd-contaminated soil. UGW-biochar amendments to the three Cd-contaminated soils resulted in an increase in the cabbage biomass in acidic soil, whereas in neutral soils, it increased in lightly contaminated soils but decreased in heavily contaminated soils. Additionally, the Cd bioaccumulation factor (BCF), translocation factor (TF), and removal efficiency (RE), as impacted by the biochar application, were calculated in the lightly Cd-contaminated soil-cabbage system. The BCF decreased from 5.84 to 3.80 as the dosage of the UGW-biochar increased from 0% to 3%, indicating that the UGW-biochar immobilized Cd and reduced its bioaccumulation in cabbage roots. Based on our investigations, UGW-biochar effectively immobilizes Cd by reducing its mobility and bioavailability in a lightly contaminated environment matrix.

Coupled sorptive and oxidative antimony(III) removal by iron-modified biochar: Mechanisms of electron-donating capacity and reactive Fe species.

Sorption and oxidation are two potential pathways for the decontamination of trivalent antimony (Sb(III))-bearing water, using iron (Fe)-modified biochar (FeBC). Here we investigated the sorption and oxidation behavior of FeBC for Sb(III) in aqueous solutions. Results revealed that Sb(III) removal by FeBC was significantly improved showing the maximum Sb(III) sorption (64.0 mg g-1). Density functional theory (DFT) calculations indicated that magnetite (Fe3O4) in FeBC offered a sorption energy of -0.22 eV, which is 5 times that of non-modified biochar. With the addition of peroxymonosulfate (PMS), the sorption of Sb(III) on FeBC was 7 times higher than that on BC, indicating the sorption capacity of FeBC for Sb(III) could be substantially increased by adding oxidizing agents. Electrochemical analysis showed that Fe modification imparted FeBC higher electron-donating capacity than that of BC (0.045 v. s. 0.023 mmol e- (g biochar)-1), which might be the reason for the strong Sb(III) oxidation (63.6%) on the surface of FeBC. This study provides new information that is key for the development of effective biochar-based composite materials for the removal of Sb(III) from drinking water and wastewater. The findings from this study have important implications for protecting human health and agriculture.

Adrenomedullin has a pivotal role in trophoblast differentiation: A promising nanotechnology-based therapeutic target for early-onset preeclampsia.

Early-onset preeclampsia (EOPE) is a severe pregnancy complication associated with defective trophoblast differentiation and functions at implantation, but manifestation of its phenotypes is in late pregnancy. There is no reliable method for early prediction and treatment of EOPE. Adrenomedullin (ADM) is an abundant placental peptide in early pregnancy. Integrated single-cell sequencing and spatial transcriptomics confirm a high ADM expression in the human villous cytotrophoblast and syncytiotrophoblast. The levels of ADM in chorionic villi and serum were lower in first-trimester pregnant women who later developed EOPE than those with normotensive pregnancy. ADM stimulates differentiation of trophoblast stem cells and trophoblast organoids in vitro. In pregnant mice, placenta-specific ADM suppression led to EOPE-like phenotypes. The EOPE-like phenotypes in a mouse PE model were reduced by a placenta-specific nanoparticle-based forced expression of ADM. Our study reveals the roles of trophoblastic ADM in placental development, EOPE pathogenesis, and its potential clinical uses.

Gut microbiota in gastrointestinal diseases during pregnancy.

Gut microbiota (GM) is a micro-ecosystem composed of all microorganisms in the human intestine. The interaction between GM and the host plays an important role in maintaining normal physiological functions in the host. Dysbiosis of the GM may cause various diseases. GM has been demonstrated to be associated with human health and disease, and changes during individual development and disease. Pregnancy is a complicated physiological process. Hormones, the immune system, metabolism, and GM undergo drastic changes during pregnancy. Gastrointestinal diseases during pregnancy, such as hepatitis, intrahepatic cholestasis of pregnancy, and pre-eclampsia, can affect both maternal and fetal health. The dysregulation of GM during pregnancy may lead to a variety of diseases, including gastrointestinal diseases. Herein, we review recent research articles on GM in pregnancy-related gastrointestinal diseases, discuss the interaction of the GM with the host under normal physiological conditions, gastrointestinal diseases, and pregnancy-specific disorders. As more attention is paid to reproductive health, the pathogenic mechanism of GM in gastrointestinal diseases during pregnancy will be further studied to provide a theoretical basis for the use of probiotics to treat these diseases.

Peripheral Blood Transcripts Predict Preoperative Obstructive Total Anomalous Pulmonary Venous Connection.

The lack of accessible noninvasive tools to examine the molecular alterations limits our understanding of the causes of total anomalous pulmonary venous connection (TAPVC), as well as the identification of effective operational strategies. Here, we consecutively enrolled peripheral leukocyte transcripts of 26 preoperative obstructive and 22 non-obstructive patients with TAPVC. Two-hundred and fifty six differentially expressed mRNA and 27 differentially expressed long noncoding RNA transcripts were dysregulated. The up-regulated mRNA was enriched in the hydrogen peroxide catabolic process, response to mechanical stimulus, neutrophil degranulation, hemostasis, response to bacterium, and the NABA CORE MATRISOME pathway, all of which are associated with the development of fibrosis. Furthermore, we constructed predictive models using multiple machine-learning algorithms and tested the performance in the validation set. The mRNA NR3C2 and lncRNA MEG3 were screened based on multiple iterations. The random forest prediction model can predict preoperative obstruction patients in the validation set with high accuracy (area under curve = 1; sensitivity = 1). These data highlight the potential of peripheral leukocyte transcripts to evaluate obstructive-related pathophysiological alterations, leading to precision healthcare solutions that could improve patient survival after surgery. It also provides a novel direction for the study of preoperative obstructive TAPVC.

Polyadenylation ligation-mediated sequencing (PALM-Seq) characterizes cell-free coding and non-coding RNAs in human biofluids.

BACKGROUND: Cell-free messenger RNA (cf-mRNA) and long non-coding RNA (cf-lncRNA) are becoming increasingly important in liquid biopsy by providing biomarkers for disease prediction, diagnosis and prognosis, but the simultaneous characterization of coding and non-coding RNAs in human biofluids remains challenging. METHODS: Here, we developed polyadenylation ligation-mediated sequencing (PALM-Seq), an RNA sequencing strategy employing treatment of RNA with T4 polynucleotide kinase to generate cell-free RNA (cfRNA) fragments with 5' phosphate and 3' hydroxyl and RNase H to deplete abundant RNAs, achieving simultaneous quantification and characterization of cfRNAs. RESULTS: Using PALM-Seq, we successfully identified well-known differentially abundant mRNA, lncRNA and microRNA in the blood plasma of pregnant women. We further characterized cfRNAs in blood plasma, saliva, urine, seminal plasma and amniotic fluid and found that the detected numbers of different RNA biotypes varied with body fluids. The profiles of cf-mRNA reflected the function of originated tissues, and immune cells significantly contributed RNA to blood plasma and saliva. Short fragments (<50 nt) of mRNA and lncRNA were major in biofluids, whereas seminal plasma and amniotic fluid tended to retain long RNA. Body fluids showed distinct preferences of pyrimidine at the 3' end and adenine at the 5' end of cf-mRNA and cf-lncRNA, which were correlated with the proportions of short fragments. CONCLUSION: Together, PALM-Seq enables a simultaneous characterization of cf-mRNA and cf-lncRNA, contributing to elucidating the biology and promoting the application of cfRNAs.

Enhanced sorption of trivalent antimony by chitosan-loaded biochar in aqueous solutions: Characterization, performance and mechanisms.

Contamination of aquatic systems by antimony (Sb) is a worldwide issue due to its risks to eco-environment and human health. Batch sorption experiments were conducted to assess the equilibrium, kinetics and thermodynamics of antimonite [Sb(III)] sorption by pristine biochar (BC) and chitosan-loaded biochar (CHBC) derived from branches of Ficus microcarpa. Results showed the successful loading of chitosan onto biochar surface, exhibiting more functional groups (e.g., CO, -NH2, and -OH). Langmuir model well described the Sb(III) sorption isotherm experimental data, and the maximum sorption capacity of Sb(III) by CH1BC (biochar loaded with chitosan at a ratio of 1:1) was 168 mg g-1, whereas for the BC it was only 10 mg g-1. X-ray photoelectron spectroscopy demonstrated that CH1BC oxidized 86% of Sb(III) to Sb(V), while BC oxidized 71% of Sb(III). Density functional theory calculations suggested that the synergistic effect of exogenous hydroxyl and inherent carbonyl contributed to the enhanced removal efficiency of Sb(III) by CHBC. Key mechanisms for Sb(III) sorption onto CHBCs included electrostatic interaction, chelation, surface complexation, π-π interaction, and hydrogen bonding. Overall, this study implies that CHBC can be a new, viable sorbent for the removal of Sb(III) from aquatic systems aiding their safe and sustainable management.