Intestinal melatonin levels and gut microbiota homeostasis are independent of the pineal gland in pigs.

Introduction: Melatonin (MEL) is a crucial neuroendocrine hormone primarily produced by the pineal gland. Pinealectomy (PINX) has been performed on an endogenous MEL deficiency model to investigate the functions of pineal MEL and its relationship with various diseases. However, the effect of PINX on the gastrointestinal tract (GIT) MEL levels and gut microbiome in pigs has not been previously reported. Methods: By using a newly established pig PINX model, we detected the levels of MEL in the GIT by liquid chromatography-tandem mass spectrometry. In addition, we examined the effects of PINX on the expression of MEL synthesis enzymes, intestinal histomorphology, and the intestinal barrier. Furthermore, 16S rRNA sequencing was performed to analyze the colonic microbiome. Results: PINX reduced serum MEL levels but did not affect GIT MEL levels. Conversely, MEL supplementation increased MEL levels in the GIT and intestinal contents. Neither PINX nor MEL supplementation had any effect on weight gain, organ coefficient, serum biochemical indexes, or MEL synthetase arylalkylamine N-acetyltransferase (AANAT) expression in the duodenum, ileum, and colon. Furthermore, no significant differences were observed in the intestinal morphology or intestinal mucosal barrier function due to the treatments. Additionally, 16S rRNA sequencing revealed that PINX had no significant impact on the composition of the intestinal microbiota. Nevertheless, MEL supplementation decreased the abundance of Fibrobacterota and increased the abundance of Actinobacteriota, Desulfobacterota, and Chloroflexi. Conclusion: We demonstrated that synthesis of MEL in the GIT is independent of the pineal gland. PINX had no influence on intestinal MEL level and microbiota composition in pigs, while exogenous MEL alters the structure of the gut microbiota.

Insomnia and anxiety among COVID-19 patients in China: the chain mediating effect of psychological capital and self-esteem.

BACKGROUND: The outbreak of Corona Virus Disease (COVID-19) in 2019 has continued until now, posing a huge threat to the public's physical and mental health, resulting in different degrees of mental health problems. As a vulnerable segment of the public, anxiety is one of the most common mental health problems among COVID-19 patients. Excessive anxiety aggravates the physical and psychological symptoms of COVID-19 patients, which is detrimental to their treatment and recovery, increases financial expenditure, affects family relations, and adds to the medical burden. OBJECTIVE: This study aimed to explore the role of psychological capital and self-esteem in the relationship between insomnia and anxiety, thereby shedding light on the mechanism of the effect of insomnia on anxiety in COVID-19 patients. METHODS: A cross-sectional study was conducted from April to May 2022 in Fangcang hospital in Shanghai, China. The self-administered questionnaires were distributed to 718 COVID-19 patients via cell phone using the Internet platform "Questionnaire Star", which included Athens Insomnia Scale, Psychological Capital Questionnaire, Self-esteem Scale, Self-Rating Anxiety Scale, gender, age, marital status, education. Data analysis was performed using descriptive analysis, independent-samples t-test, one-way analysis of variance, Pearson correlation analysis, ordinary least-squares regression, and bootstrap method. RESULTS: Education background had significant impact on anxiety in COVID-19 patients (F = 7.70, P < 0.001). Insomnia, psychological capital, self-esteem and anxiety were significantly correlated, respectively (P < 0.001). And Regression analysis showed that insomnia had a direct negative predictive effect on psychological capital (ß = -0.70, P < 0.001) and self-esteem (ß = -0.13, P < 0.001). Psychological capital had a direct positive predictive effect on self-esteem (ß = 0.12, P < 0.001). Insomnia had a direct positive predictive effect on anxiety (ß = 0.61, P < 0.001). Both psychological capital and self-esteem had significant negative predictive effects on anxiety (ß = -0.06, P < 0.05; ß = -0.72, P < 0.001). The results showed that the mediating effect of psychological capital and self-esteem was significant, and the mediating effect value was 0.21. First, the indirect effect consisting of insomnia - psychological capital - anxiety was 0.04, showing that psychological capital had a significant mediating effect. Second, the indirect effect consisting of insomnia-self-esteem-anxiety had a value of 0.10, indicating that self-esteem had a significant mediating effect. Third, the indirect effect consisting of insomnia-psychological capital-self-esteem-anxiety had a value of 0.06, suggesting that psychological capital and self-esteem had a significant chain mediating effect between insomnia and anxiety. CONCLUSIONS: Insomnia had a significant positive predictive effect on anxiety. Insomnia was first associated with a decrease in psychological capital, followed by a sequential decrease in self-esteem, which in turn was associated with increased anxiety symptoms in COVID-19 patients. Therefore, focusing on improving the psychological capital and self-esteem of patients can help alleviate the anxiety caused by insomnia in COVID-19 patients. It is recommended that patients and health care professionals increase the psychological capital and Self-esteem of COVID-19 patients through various methods to counter the effects of insomnia on anxiety.

Performance Enhancement of Silicone Rubber Using Superhydrophobic Silica Aerogel with Robust Nanonetwork Structure and Outstanding Interfacial Effect.

The application of high-performance rubber nanocomposites has attracted wide attention, but its development is limited by the imbalance of interface and network effects caused by fillers. Herein, an ultrastrong polymer nanocomposite is successfully designed by introducing a superhydrophobic and mesoporous silica aerogel (HSA) as the filler to poly(methyl vinyl phenyl) siloxane (PVMQ), which increased the PVMQ elongation at break (∼690.1%) by ∼9.3 times and the strength at break (∼6.6 MPa) by ∼24.3 times. Furthermore, HSA/PVMQ with a high dynamic storage modulus (G'0) of ∼12.2 MPa and high Payne effect (ΔG') of ∼9.4 MPa is simultaneously achieved, which is equivalent to 2-3 times that of commercial fumed silica reinforced PVMQ. The superior performance is attributed to the filler-rubber interfacial interaction and the robust filler-rubber entanglement network which is observed by scanning electron microscopy. When the HSA-PVMQ entanglement network is subjected to external stress, both the HSA and bound-PVMQ chains are synergistically involved in resisting structural evolution, resulting in the maximized energy dissipation and deformation resistance through the desorption of the polymer chain and the slip/interpenetrating of the exchange hydrogen bond pairs. Hence, highly aggregated nanoporous silica aerogels may soon be widely used in the application of reinforced silicone rubber or other polymers shortly.

Combined Use of Tocilizumab and Mesenchymal Stem Cells Attenuate the Development of an Anti-HLA-A2.1 Antibody in a Highly Sensitized Mouse Model.

Sensitization to HLA can result in allograft loss for kidney transplantation (KT) patients. Therefore, it is required to develop an appropriate desensitization (DSZ) technique to remove HLA-donor-specific anti-HLA antibody (DSA) before KT. The aim of this research was to investigate whether combined use of the IL-6 receptor-blocking antibody, tocilizumab (TCZ), and bone-marrow-derived mesenchymal stem cells (BM-MSCs) could attenuate humoral immune responses in an allo-sensitized mouse model developed using HLA.A2 transgenic mice. Wild-type C57BL/6 mice were sensitized with skin allografts from C57BL/6-Tg (HLA-A2.1)1Enge/J mice and treated with TCZ, BM-MSC, or both TCZ and BM-MSC. We compared HLA.A2-specific IgG levels and subsets of T cells and B cells using flow cytometry among groups. HLA.A2-specific IgG level was decreased in all treated groups in comparison with that in the allo-sensitized control (Allo-CONT) group. Its decrease was the most significant in the TCZ + BM-MSC group. Regarding the B cell subset, combined use of TCZ and BM-MSC increased proportions of pre-pro B cells but decreased proportions of mature B cells in BM (p < 0.05 vs. control). In the spleen, an increase in transitional memory was observed with a significant decrease in marginal, follicular, and long-lived plasma B cells (p < 0.05 vs. control) in the TCZ + BM-MSC group. In T cell subsets, Th2 and Th17 cells were significantly decreased, but Treg cells were significantly increased in the TCZ+BM-MSC group compared to those in the Allo-CONT group in the spleen. Regarding RNA levels, IL-10 and Foxp3 showed increased expression, whereas IL-23 and IFN-γ showed decreased expression in the TCZ + BM-MSC group. In conclusion, combined use of TCZ and BM-MSC can inhibit B cell maturation and up-regulate Treg cells, finally resulting in the reduction of HLA.A2-specific IgG in a highly sensitized mouse model. This study suggests that the combined use of TCZ and BM-MSC can be proposed as a novel strategy in a desensitization protocol for highly sensitized patients.

Highly sensitive fluorescent explosives detection via SERS: based on fluorescence quenching of graphene oxide@Ag composite aerogels.

High fluorescence background poses a substantial challenge to surface-enhanced Raman scattering (SERS), thereby limiting its broader applicability across diverse domains. In this work, silver nanoparticle (Ag NP)-loaded graphene oxide aerogel nanomaterials (GO-Ag ANM) were prepared for sensitive SERS detection of fluorescent explosive 2,4,8,10-tetranitrobenzo-1,3a,6,6a-tetraazapentaenopyridine (BPTAP) by a fluorescence quenching strategy. By harnessing the fluorescence quenching properties of graphene and the localized surface plasmon resonance of silver nanoparticles, the synthesized aerogels exhibited effective fluorescence quenching and Raman enhancement capabilities when employed for BPTAP analysis with 532 nm laser excitation. Significantly, precise control over the loading quantity of silver nanoparticles (Ag NPs) resulted in the remarkable sensitivity of the surface-enhanced Raman scattering (SERS) effect. This method allowed for the detection of fluorescent explosive BPTAP at an extraordinarily low concentration of 1 × 10-7 M. Furthermore, the approach also demonstrated excellent detection capabilities for the dyes R6G, CV, and RhB. This study offers valuable insights for the sensitive detection of fluorescent molecules.

Splicing factor SRSF1 is essential for homing of precursor spermatogonial stem cells in mice.

Spermatogonial stem cells (SSCs) are essential for continuous spermatogenesis and male fertility. The underlying mechanisms of alternative splicing (AS) in mouse SSCs are still largely unclear. We demonstrated that SRSF1 is essential for gene expression and splicing in mouse SSCs. Crosslinking immunoprecipitation and sequencing data revealed that spermatogonia-related genes (e.g. Plzf, Id4, Setdb1, Stra8, Tial1/Tiar, Bcas2, Ddx5, Srsf10, Uhrf1, and Bud31) were bound by SRSF1 in the mouse testes. Specific deletion of Srsf1 in mouse germ cells impairs homing of precursor SSCs leading to male infertility. Whole-mount staining data showed the absence of germ cells in the testes of adult conditional knockout (cKO) mice, which indicates Sertoli cell-only syndrome in cKO mice. The expression of spermatogonia-related genes (e.g. Gfra1, Pou5f1, Plzf, Dnd1, Stra8, and Taf4b) was significantly reduced in the testes of cKO mice. Moreover, multiomics analysis suggests that SRSF1 may affect survival of spermatogonia by directly binding and regulating Tial1/Tiar expression through AS. In addition, immunoprecipitation mass spectrometry and co-immunoprecipitation data showed that SRSF1 interacts with RNA splicing-related proteins (e.g. SART1, RBM15, and SRSF10). Collectively, our data reveal the critical role of SRSF1 in spermatogonia survival, which may provide a framework to elucidate the molecular mechanisms of the posttranscriptional network underlying homing of precursor SSCs.

CK1α deficiency impairs mouse uterine adenogenesis by inducing epithelial cell apoptosis through GSK3ß pathway and inhibiting Foxa2 expression through p53 pathway†.

Uterine glands and their secretions are crucial for conceptus survival and implantation in rodents and humans. In mice, the development of uterine gland known as adenogenesis occurs after birth, whereas the adenogenesis in humans initiates from fetal life and completed at puberty. Uterine adenogenesis involves dynamic epithelial cell proliferation, differentiation, and apoptosis. However, it is largely unexplored about the mechanisms governing adenogenesis. CK1α plays important roles in regulating cell division, differentiation, and death, but it is unknown whether CK1α affects adenogenesis. In the current study, uterus-specific CK1α knockout female mice (Csnk1a1d/d) were infertile resulted from lack of uterine glands. Subsequent analysis revealed that CK1α deletion induced massive apoptosis in uterine epithelium by activating GSK3ß, which was confirmed by injections of GSK3ß inhibitor SB216763 to Csnk1a1d/d females, and the co-treatment of SB216763 and CK1 inhibitor d4476 on cultured epithelial cells. Another important finding was that our results revealed CK1α deficiency activated p53, which then blocked the expression of Foxa2, an important factor for glandular epithelium development and function. This was confirmed by that Foxa2 expression level was elevated in p53 inhibitor pifithrin-α injected Csnk1a1d/d mouse uterus and in vitro dual-luciferase reporter assay between p53 and Foxa2. Collectively, these studies reveal that CK1α is a novel factor regulating uterine adenogenesis by inhibiting epithelial cell apoptosis through GSK3ß pathway and regulating Foxa2 expression through p53 pathway. Uncovering the mechanisms of uterine adenogenesis is expected to improve pregnancy success in humans and other mammals.

PGF2alpha Inhibits 20alpha-HSD Expression by Suppressing CK1alpha-induced ERK and SP1 Activation in the Corpus Luteum of Pregnant Mice.

Prostaglandin F2α (PGF2α) is a luteolytic hormone that promotes parturition in mammals at the end of pregnancy by reducing progesterone secretion from the corpus luteum (CL). In rodents and primates, PGF2α rapidly converts progesterone to 20α-hydroxyprogesterone (20α-OHP) by promoting 20α-hydroxysteroid dehydrogenase (20α-HSD) expression. However, the specific mechanism of 20α-HSD regulation by PGF2α remains unclear. Casein Kinase 1α (CK1α) is a CK1 family member that regulates a variety of physiological functions, including reproductive development. Here, we investigated the effects of CK1α on pregnancy in female mice. Our experiments showed that CK1α is expressed in mouse CL, and its inhibition enhanced progesterone metabolism, decreased progesterone levels, and affected mouse embryo implantation. Further, CK1α mediated the effect of PGF2α on 20α-HSD in mouse luteal cells in vitro. Our results are the first to show that CK1α affects the 20α-HSD mRNA level by affecting the ERK signalling pathway to regulate the expression of the transcription factor SP1. These findings improve our understanding of PGF2α regulation of 20α-HSD.

Nucleic-acid-triggered NADase activation of a short prokaryotic Argonaute.

Argonaute (Ago) proteins mediate RNA- or DNA-guided inhibition of nucleic acids1,2. Although the mechanisms used by eukaryotic Ago proteins and long prokaryotic Ago proteins (pAgos) are known, that used by short pAgos remains elusive. Here we determined the cryo-electron microscopy structures of a short pAgo and the associated TIR-APAZ proteins (SPARTA) from Crenotalea thermophila (Crt): a free-state Crt-SPARTA; a guide RNA-target DNA-loaded Crt-SPARTA; two Crt-SPARTA dimers with distinct TIR organization; and a Crt-SPARTA tetramer. These structures reveal that Crt-SPARTA is composed of a bilobal-fold Ago lobe that connects with a TIR lobe. Whereas the Crt-Ago contains a MID and a PIWI domain, Crt-TIR-APAZ has a TIR domain, an N-like domain, a linker domain and a trigger domain. The bound RNA-DNA duplex adopts a B-form conformation that is recognized by base-specific contacts. Nucleic acid binding causes conformational changes because the trigger domain acts as a 'roadblock' that prevents the guide RNA 5' ends and the target DNA 3' ends from reaching their canonical pockets; this disorders the MID domain and promotes Crt-SPARTA dimerization. Two RNA-DNA-loaded Crt-SPARTA dimers form a tetramer through their TIR domains. Four Crt-TIR domains assemble into two parallel head-to-tail-organized TIR dimers, indicating an NADase-active conformation, which is supported by our mutagenesis study. Our results reveal the structural basis of short-pAgo-mediated defence against invading nucleic acids, and provide insights for optimizing the detection of SPARTA-based programmable DNA sequences.

Taurine promotes insulin synthesis by enhancing Isl-1 expression through miR-7a/RAF1/ERK1/2 pathway.

It has been well established that the circulating taurine affects the insulin synthesis in pancreatic islet ß-cells, whereas miR-7a and LIM-homeodomain transcription factor Isl-1 are important intracellular factors regulating insulin transcription and synthesis. However, it still remains unknown whether taurine regulates insulin synthesis by affecting miR-7a and/or Isl-1 expressions in mouse pancreatic islet ß-cells. The present study was thus proposed to identify the effects of taurine on the expressions of miR-7a and/or Isl-1 and their relations to insulin synthesis in mouse pancreatic islet ß-cells by using miR-7a2 knockout (KO) and taurine transporter (TauT) KO mouse models and the related in vitro experiments. The results demonstrated that taurine supplement significantly decreased the pancreas miR-7a expression, but sharply upregulated the pancreas Isl-1 and insulin expressions, and serum insulin levels. However, the enhanced effects of taurine on Isl-1 expression and insulin synthesis were mitigated in the TauT KO and miR-7a2 KO mice. In addition, our results confirmed that taurine markedly increased pancreas RAF1 and ERK1/2 expressions. Collectively, the present study firstly demonstrates that taurine regulates insulin synthesis through TauT/miR-7a/RAF1/ERK1/2/Isl-1 signaling pathway, which are crucial for our understanding the mechanisms of taurine affecting insulin synthesis, and also potential for establishing the therapeutic strategies for diabetes and the diseases related to metabolism.

Modulating Electronic Structure and Atomic Insights into the Novel Hierarchically Porous PdCuFe Trimetallic Alloy Aerogel for Efficient Oxygen Reduction.

The high cost of noble Pd/Pt required for the oxygen reduction reaction (ORR) in the cathode restricts the wide applications of fuel cells. In this study, the synthesis of a novel Pd3 CuFe0.5 aerogel electrocatalyst is successfully demonstrated using self-assembly and lyophilization techniques, employing a mild reducing agent. The resulting aerogel electrocatalyst exhibits a distinctive 3D network structure, possessing a substantial BET-specific surface area of 75.19 m2  g-1 . It is worth noting that the optimized Pd3 CuFe0.5 aerogel demonstrates exceptional ORR performance with a high half-wave potential of 0.92 V versus RHE, a significant limiting current density of 7.6 mA cm-2 , and the excellent electrocatalytic stability, superior to the reported noble metal electrocatalysts, with the ORR activity decays only 4.9% after 16 000 s. In addition, the Pd3 CuFe0.5 aerogel electrocatalyst shows superior cycling stability for ≈120 h at a charge/discharge current density of 10 mA cm-2 , indicating its promising application in fuel cells. Furthermore, the resulting composite aerogel possesses excellent hydrogen evolution reaction and ethanol oxidation reaction activity. The density functional theory calculations show that the partial oxidation of Pd3 CuFe0.5 aerogel leads to a negative shift of the d-band center, which energetically optimizes the binding strength of *O intermediates, therefore accelerating the ORR activity.

Facile Preparation of a Novel HfC Aerogel with Low Thermal Conductivity and Excellent Mechanical Properties.

Aerogels emerge as captivating contenders within the realm of high-temperature thermal resistance and thermal insulation. Nevertheless, their practical applications are usually constrained by their inherent brittleness when subjected to rigorous conditions. Herein, employing hafnium dichloride oxide octahydrate (HfOCl2·8H2O) as the hafnium source and resorcinol-formaldehyde (RF) as the carbon precursor, hafnium carbide (HfC) aerogels are fabricated via the sol-gel method complemented with carbothermal reduction reaction. Investigations are conducted into the effects of various molar ratios, duration, and temperatures of calcination on the microstructural features and physico-chemical characteristics of the as-prepared HfC aerogel. The aerogel shows a high BET-specific surface area (601.02 m2/g), which is much larger than those of previously reported aerogels. Furthermore, the HfC aerogel exhibits a low thermal conductivity of 0.053 W/(m·K) and a compressive strength of up to 6.12 MPa after carbothermal reduction at 1500 °C. These excellent thermal insulation and mechanical properties ensure it is ideal for the utilization of high-temperature thermal resistance and thermal insulation in the fields of aerospace.

Genomic prediction of pig growth traits based on machine learning.

This study aimed to assess and compare the performance of different machine learning models in predicting selected pig growth traits and genomic estimated breeding values (GEBV) using automated machine learning, with the goal of optimizing whole-genome evaluation methods in pig breeding. The research employed genomic information, pedigree matrices, fixed effects, and phenotype data from 9968 pigs across multiple companies to derive four optimal machine learning models: deep learning (DL), random forest (RF), gradient boosting machine (GBM), and extreme gradient boosting (XGB). Through 10-fold cross-validation, predictions were made for GEBV and phenotypes of pigs reaching weight milestones (100 kg and 115 kg) with adjustments for backfat and days to weight. The findings indicated that machine learning models exhibited higher accuracy in predicting GEBV compared to phenotypic traits. Notably, GBM demonstrated superior GEBV prediction accuracy, with values of 0.683, 0.710, 0.866, and 0.871 for B100, B115, D100, and D115, respectively, slightly outperforming other methods. In phenotype prediction, GBM emerged as the best-performing model for pigs with B100, B115, D100, and D115 traits, achieving prediction accuracies of 0.547, followed by DL at 0.547, and then XGB with accuracies of 0.672 and 0.670. In terms of model training time, RF required the most time, while GBM and DL fell in between, and XGB demonstrated the shortest training time. In summary, machine learning models obtained through automated techniques exhibited higher GEBV prediction accuracy compared to phenotypic traits. GBM emerged as the overall top performer in terms of prediction accuracy and training time efficiency, while XGB demonstrated the ability to train accurate prediction models within a short timeframe. RF, on the other hand, had longer training times and insufficient accuracy, rendering it unsuitable for predicting pig growth traits and GEBV.

BCAS2 Participates in Insulin Synthesis and Secretion via mRNA Alternative Splicing in Mice.

Insulin secreted by pancreatic ß cells is essential for maintaining blood glucose levels. Diabetes is caused primarily by a loss of ß cells or impairment of ß-cell function. A previous whole-transcriptome analysis of islets from a type 2 diabetes group and a control group showed that a splicing disorder occurred in approximately 25% of splicing events. Breast carcinoma amplified sequence 2 (BCAS2) is a spliceosome component whose function in islet ß cells is unclear. Here, we report that knockdown of Bcas2 decreased glucose- and KCl-stimulated insulin secretion in the NIT-1 cell line. Pancreas weight, glucose tolerance, and insulin sensitivity were measured in normal chow-fed Bcas2 f/f-ßKO mice, and ß-cell mass and islet size were analyzed by immunohistochemistry. Glucose intolerance developed in Bcas2 f/f-ßKO mice, but there were no significant differences in pancreas weight, insulin sensitivity, ß-cell mass, or islet size. Furthermore, observation of glucose-stimulated insulin secretion and insulin secretion granules in normal chow-fed mice revealed that the insulin level in serum and the number of insulin secretion granules were decreased in Bcas2 f/f-ßKO mice. These differences were related to abnormal splicing of Syt7 and Tcf7l2 pre-mRNA. Taken together, these results demonstrate that BCAS2 is involved in alternative splicing during insulin synthesis and secretion.

Identification and Quantification of the Main Psychoactive Ingredients of Cannabis in Urine Using Excitation-Eemission Matrix Fluorescence Coupled with Parallel Factor Analysis.

Cannabis is the most prevalent abused substance after alcohol, and its consumption severely harms human health and thus adversely impacts society. The identification and quantification of cannabis in urine play important roles in practical forensics. Excitation-emission matrix (EEM) fluorescence spectroscopy coupled with parallel factor (PARAFAC) analysis was developed to identify and quantify the four main ingredients of cannabis in urine samples. The main ingredients of cannabis including Δ-9-tetrahydrocannabinol (THC), cannabidiol, cannabinol, and tetrahydrocannabinolic acid (THC-COOH) exhibited diverse fluorescence characteristics, and the concentrations of these compounds depicted a positive linear relationship with the fluorescence intensity at the ng/mL level. The EEM/PARAFAC method adequately characterized and discriminated the four ingredients in calibration and prediction samples with a low root-mean-square error of prediction (RMSEP; 0.03-0.07 µg/mL) and limit of quantitation (LOQ; 0.26-0.71 µg/mL). The prediction results of the EEM/PARAFAC method well correlated with that of GC-MS with a low RMSEP range (0.01-0.05 µg/mL) and LOQ range (0.07-0.44 µg/mL) in urine samples. The EEM spectroscopic investigation coupled with the PARAFAC algorithm results in an organic, solvent-less, fast, reliable tool to perform accurate and rapid screening of cannabis abusers.

Modeling of FAN1-Deficient Kidney Disease Using a Human Induced Pluripotent Stem Cell-Derived Kidney Organoid System.

Karyomegalic interstitial nephritis (KIN) is a genetic kidney disease caused by mutations in the FANCD2/FANCI-Associated Nuclease 1 (FAN1) gene on 15q13.3, which results in karyomegaly and fibrosis of kidney cells through the incomplete repair of DNA damage. The aim of this study was to explore the possibility of using a human induced pluripotent stem cell (hiPSC)-derived kidney organoid system for modeling FAN1-deficient kidney disease, also known as KIN. We generated kidney organoids using WTC-11 (wild-type) hiPSCs and FAN1-mutant hiPSCs which include KIN patient-derived hiPSCs and FAN1-edited hiPSCs (WTC-11 FAN1+/-), created using the CRISPR/Cas9 system in WTC-11-hiPSCs. Kidney organoids from each group were treated with 20 nM of mitomycin C (MMC) for 24 or 48 h, and the expression levels of Ki67 and H2A histone family member X (H2A.X) were analyzed to detect DNA damage and assess the viability of cells within the kidney organoids. Both WTC-11-hiPSCs and FAN1-mutant hiPSCs were successfully differentiated into kidney organoids without structural deformities. MMC treatment for 48 h significantly increased the expression of DNA damage markers, while cell viability in both FAN1-mutant kidney organoids was decreased. However, these findings were observed in WTC-11-kidney organoids. These results suggest that FAN1-mutant kidney organoids can recapitulate the phenotype of FAN1-deficient kidney disease.

Identification and classification of ATS in oral fluid based on Ag nanoassemblies on Si surface doped with Au nanobipyramids.

Herein, a novel Ag NP substrate doped with Au nanobipyramids was designed and fabricated via a convenient procedure of galvanic reaction for the identification and classification of amphetamine-type stimulants (ATS) in oral fluids in combination with surface enhanced Raman scattering (SERS). The substrate was shown to have a three-dimensional nanostructure, high SERS activity, and good stability. In combination with SERS, the Ag NP substrate doped with Au nanobipyramids was able to detect ultra-low traces of ATS, including amphetamine, methylamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA), and 3,4-methylenedioxymethylamphetamine (MDMA) in oral fluid with limit of detection (LOD) and limit of determination quantitation (LOQ) as low as 10-9 mg/mL, which is much better than the current spectroscopic techniques. The equations between concentration and peaks intensity for quantitative analysis displied good doublelogarithmic linear relations and reliability figures of merit at nanogram concentration level in compartion with GC-MS method. The approach can be broadly applied to the ultra-low trace detection of ATS in oral fluid and would be particularly useful for the analyses of nitrogenous organic compounds.

A novel magnetic Fe3O4/cellulose nanofiber/polyethyleneimine/thiol-modified montmorillonite aerogel for efficient removal of heavy metal ions: Adsorption behavior and mechanism study.

To efficiently remove heavy metals from wastewater, designing an adsorbent with high adsorption capacity and ease of recovery is necessary. This paper presents a novel magnetic hybridized aerogel, Fe3O4/cellulose nanofiber/polyethyleneimine/thiol-modified montmorillonite (Fe3O4/CNF/PEI/SHMMT), and explores its adsorption performance and mechanism for Pb2+, Cu2+, and Cd2+ in aqueous solutions. The hybrid aerogel has a slit-like porous structure and numerous exposed active sites, which facilitates the uptake of metal ions by adsorption. Pb2+, Cu2+, and Cd2+ adsorption by the hybridized aerogel followed the second-order kinetics and the Langmuir isotherm model, the maximum adsorption of Pb2+, Cu2+, and Cd2+ at 25 °C, pH = 6, 800 mg/L was 429.18, 381.68 and 299.40 mg/g, respectively. The adsorption process was primarily attributed to monolayer chemical adsorption, a spontaneous heat-absorption reaction. FTIR, XPS and DFT studies confirmed that the adsorption mechanisms of Fe3O4/CNF/PEI/SHMMT on Pb2+, Cu2+, and Cd2+ were mainly chelation, coordination, and ion exchange. The lowest adsorption energy of Pb2+ on the hybrid aerogel was calculated to be -2.37 Ha by DFT, which indicates that the sample has higher adsorption affinity and preferential selectivity for Pb2+. After 5 cycles, the adsorption efficiency of the aerogel was still >85 %. The incorporation of Fe3O4 improved the mechanical properties of the aerogel. The Fe3O4/CNF/PEI/SHMMT has fast magnetic responsiveness, and it is easy to be separated and recovered after adsorption, which is a promising potential for the treatment of heavy metal ions.

Neuroligin-3 activates Akt-dependent Nrf2 cascade to protect osteoblasts from oxidative stress.

Excessive oxidative stress will cause significant injury to osteoblasts, serving as one major pathological mechanism of osteoporosis. Neuroligin-3 (NLGN3) is a postsynaptic cell adhesion protein and is expressed in the bone. We here explored its potential activity against hydrogen peroxide (H2O2)-induced oxidative injury in cultured osteoblasts. In primary murine and human osteoblasts, NLGN3 stimulation dose-dependently induced Akt, Erk1/2 and S6K activation. NLGN3 pretreatment ameliorated H2O2-induced cytotoxicity and death in osteoblasts. Moreover, H2O2-induced reactive oxygen species (ROS) production and oxidative injury were alleviated with NLGN3 pretreatment in cultured osteoblasts. Further studies showed that NLGN3 activated Nrf2 signaling cascade and induced Nrf2 protein Serine-40 phosphorylation, Keap1-Nrf2 dissociation, Nrf2 protein stabilization and nuclear translocation in osteoblasts. NLGN3 also increased antioxidant response element (ARE) activity and induced expression of Nrf2-ARE-dependent genes (HO1, GCLC and NQO1) in osteoblasts. Moreover NLGN3 mitigated osteoblast oxidative injury by dexamethasone or sodium fluoride (NaF). Nrf2 cascade activation is essential for NLGN3-induced cytoprotective activity in osteoblasts. Nrf2 shRNA or knockout (KO) abolished NLGN3-induced osteoblast cytoprotection against H2O2. Contrarily forced Nrf2 cascade activation by Keap1 KO mimicked NLGN3-induced anti-oxidative activity in murine osteoblasts. Importantly, NLGN3-induced Serine-40 phosphorylation and Nrf2 cascade activation were blocked by an Akt inhibitor MK-2206 or by Akt1 shRNA. Importantly, Akt inhibition, Akt1 silencing or Nrf2 S40T mutation largely inhibited NLGN3-induced osteoblast cytoprotection against H2O2. At last, we showed that NLGN3 mRNA and protein expression was significantly downregulated in necrotic bone tissues of dexamethasone-taken patients. Taken together, NLGN3 activated Akt-dependent Nrf2 cascade to protect osteoblasts from oxidative stress.

Low complexity joint clock recovery and adaptive equalization based on a baud-rate timing phase error detector for short-reach digital coherent transmission.

Digital coherent receivers adopting joint clock recovery and adaptive equalization (JCA) can avoid failures of the adaptive equalizer (AEQ) or clock recovery algorithm (CRA) due to clock asynchrony and chromatic dispersion (CD). But in the previous JCA scheme, the AEQ has a heavy computational load as it has to generate two samples per symbol (SPS) for the subsequent timing phase error detector (TPED) which is the core of the CRA. Furthermore, the previous JCA scheme cannot compensate for receiver skew or accommodate Nyquist signals with small roll-off factors (ROFs). These shortcomings hinder its practical applications in ultrahigh-speed short-reach coherent transmission requiring low power consumption, high spectral efficiency, whilst being sensitive to receiver skew. To solve this problem, we propose a new JCA scheme by integrating a two-section real-valued (RV) AEQ with an all-digital feedback CRA based on a baud-rate TPED versatile for different ROFs. Experiments for 61-GBaud dual-polarization (DP) Nyquist 16QAM signals with an ROF of 0.01 show that, compared with the previous JCA scheme, the proposed scheme can reduce the AEQ computational load by about 70% for 10-km transmission, whilst improving the receiver sensitivity by more than 1.7 dB for a receiver skew of 1.5 ps. As far as we know, the proposed JCA scheme is the most comprehensive and efficient solution for ultrahigh-speed short-reach coherent transmission where CD, receiver skew, clock asynchrony, and Nyquist signals with small ROFs have to be dealt with.