Chlorogenic acid improves intestinal morphology by enhancing intestinal stem-cell activity.

BACKGROUND: Chlorogenic acid (CGA), as one of the most abundant naturally occurring phenolic acids, has been documented to be beneficial for intestinal health. However, the underlying mechanism is still not fully understood. The adult intestinal stem cell is the critical driver of epithelial homeostasis and regeneration. RESULTS: This study hypothesized that CGA exerted intestinal health effects by modulating intestinal stem-cell functions. Lgr5-EGFP mice were treated for 14 days, and intestinal organoids derived from these mice were treated for 3 days, using CGA solution. In comparison with the control group, CGA treatment increased intestinal villous height and crypt depth in mice and augmented the area expansion and the number of budding intestinal organoids. Quantitative polymerase chain reaction (qPCR) analysis revealed that CGA treatment significantly increased the expression of genes coding intestinal stem-cell markers in intestinal tissue and organoids, and upregulated the expression of genes coding secretory cell lineages and enterocytes, although not statistically significantly. Fluorescence-activated cell-sorting analysis further confirmed that CGA augmented the number of stem cells. 5-Ethynyl-2'-deoxyuridine (EdU) incorporation and Ki67 immunostaining results also demonstrated that CGA treatment enhanced intestinal stem-cell proliferation. CONCLUSION: Altogether, our findings indicate that CGA could activate intestinal stem-cell and epithelial regeneration, which could contribute to the improvement of intestinal morphology or organoid growth of mice. This highlights a promising mechanism for CGA as an excellent candidate for the formulation of dietary supplements and functional foods for intestinal protection. © 2023 Society of Chemical Industry.

Downregulated circ_0001681 Predicts the Occurrence and Malignant Development of Lung Carcinoma via Negatively Modulating miR-567.

OBJECTIVE: The screening and monitoring of lung adenocarcinoma (LUAD) is critical for its therapeutic efficiency. This study explored a novel biomarker of LUAD from the GSE146689 dataset and evaluated its function aiming to provide a potential therapeutic target for LUAD. METHODS: There were 126 LUAD patients, 75 benign lung disease patients, and 57 healthy individuals enrolled in this study. circ_0001681 levels were evaluated by PCR, and its significance in the diagnosis and prognosis of LUAD was assessed by ROC and Cox regression analysis. The effect of circ_0001681 on LUAD cells was investigated by CCK8 and transwell assays. RESULTS: Reduced circ_0001681 was observed in LUAD patients relative to benign lung disease patients and healthy individuals, which could discriminate LUAD patients. circ_0001681 downregulation was closely associated with the severity and malignancy of LUAD patients, and it also indicated the adverse prognosis of LUAD. In mechanism, circ_0001681 could negatively regulate miR-567, which was found to mediate the inhibitory effect of circ_0001681 on LUAD cell proliferation and motility. CONCLUSION: Decreased circ_0001681 in LUAD served as a biomarker in tumor occurrence and development, which can be included in the formulation and adjustment of LUAD therapeutic strategy.

The mTOR/PGC-1α/SIRT3 Pathway Drives Reductive Glutamine Metabolism to Reduce Oxidative Stress Caused by ISKNV in CPB Cells.

Under oxidative stress, viruses prefer glycolysis as an ATP source, and glutamine is required as an anaplerotic substrate to replenish the TCA cycle. Infectious spleen and kidney necrosis virus (ISKNV) induces reductive glutamine metabolism in the host cells. Here we report that ISKNV infection the increased NAD+/NADH ratio and the gene expression of glutaminase 1 (GLS1), glutamate dehydrogenase (GDH), and isocitrate dehydrogenase (IDH2) resulted in the phosphorylation and activation of mammalian target of rapamycin (mTOR) in CPB cells. Inhibition of mTOR signaling attenuates ISKNV-induced the upregulation of GLS1, GDH, and IDH2 genes expression, and exhibits significant antiviral activity. Moreover, the expression of silent information regulation 2 homolog 3 (SIRT3) in mRNA level is increased to enhance the reductive glutamine metabolism in ISKNV-infected cells. And those were verified by the expression levels of metabolic genes and the activities of metabolic enzymes in SIRT3-overexpressed or SIRT3-knocked down cells. Remarkably, activation of mTOR signaling upregulates the expression of the peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) gene, leading to increased expression of SIRT3 and metabolic genes. These results indicate that mTOR signaling manipulates reductive glutamine metabolism in ISKNV-infected cells through PGC-1α-dependent regulation of SIRT3. Our findings reveal new insights on ISKNV-host interactions and will contribute new cellular targets to antiviral therapy. IMPORTANCE Infectious spleen and kidney necrosis virus (ISKNV) is the causative agent of farmed fish disease that has caused huge economic losses in fresh and marine fish aquaculture. The redox state of cells is shaped by virus into a favorable microenvironment for virus replication and proliferation. Our previous study demonstrated that ISKNV replication induced glutamine metabolism reprogramming, and it is necessary for the ISKNV multiplication. In this study, the mechanistic link between the mTOR/PGC-1α/SIRT3 pathway and reductive glutamine metabolism in the ISKNV-infected cells was provided, which will contribute new insights into the pathogenesis of ISKNV and antiviral treatment strategies.

No Exchange of Picornaviruses in Vietnam between Humans and Animals in a High-Risk Cohort with Close Contact despite High Prevalence and Diversity.

Hospital-based and community-based 'high-risk cohort' studies investigating humans at risk of zoonotic infection due to occupational or residential exposure to animals were conducted in Vietnam, with diverse viruses identified from faecal samples collected from humans, domestic and wild animals. In this study, we focus on the positive-sense RNA virus family Picornaviridae, investigating the prevalence, diversity, and potential for cross-species transmission. Through metagenomic sequencing, we found picornavirus contigs in 23% of samples, belonging to 15 picornavirus genera. Prevalence was highest in bats (67%) while diversity was highest in rats (nine genera). In addition, 22% of the contigs were derived from novel viruses: Twelve phylogenetically distinct clusters were observed in rats of which seven belong to novel species or types in the genera Hunnivirus, Parechovirus, Cardiovirus, Mosavirus and Mupivirus; four distinct clusters were found in bats, belonging to one novel parechovirus species and one related to an unclassified picornavirus. There was no evidence for zoonotic transmission in our data. Our study provides an improved knowledge of the diversity and prevalence of picornaviruses, including a variety of novel picornaviruses in rats and bats. We highlight the importance of monitoring the human-animal interface for possible spill-over events.

Enhanced performance of an acetone gas sensor based on Ag-LaFeO3 molecular imprinted polymers and carbon nanotubes composite.

High performance acetone gas sensors were fabricated with molecular imprinted polymers of Ag-LaFeO3 (ALFOMMIPs) and multi walled carbon nanotubes (CNTs) composite using the microwave assisted sol-gel method. The crystalline structure, functional groups, grain size and surface appearance of the synthesized materials were analyzed via different characterization techniques and the gas responses of the samples were examined. The detailed acetone gas sensing tests and analysis revealed that the CNTs and ALFOMIPs nanocomposite (CNT/ALFOMIP) sample possessed a higher response than that of the ALFOMIPs sample. Where 0.75 wt% CNTs were added into the ALFOMIPs (0.75% CNT/ALFOMIP nanocomposite) sensor, an excellent gas sensing performance was exhibited. The response of this sensor was up to 59 for 5 ppm acetone vapors and the response and recovery times were 58 and 33 s at low working temperature of 86 °C, respectively. In addition, it had the best selectivity only to acetone vapors due to the use of the molecular imprinting technique.

Design of ultrasensitive Ag-LaFeO3 methanol gas sensor based on quasi molecular imprinting technology.

An ultrasensitive methanol gas sensing device based on the quasi-molecular imprinting technology (quasi-MIT) is studied in this work. We applied the sol-gel method (ALS denotes Ag-LaFeO3 prepared by the sol-gel method) and combustion synthesis (ALC denotes Ag-LaFeO3 prepared by combustion synthesis) to prepare Ag-LaFeO3 based sensors. The morphologies and structures of the Ag-LaFeO3 materials were examined via various detection techniques. The ALSM and ALCM sensor (ALSM and ALCM denotes the devices prepared by coating the ALS and ALC materials with methanol, respectively) fabricated using the sol-gel method and combustion synthesis combined with quasi-MIT exhibit good gas sensing properties to methanol, in contrast with the two devices (ALSW and ALCW denote the devices prepared for coating the ALS and ALC materials with water, respectively) without the use of quasi-MIT. The results show that quasi-MIT introduced the target gas in the fabrication process of the device, playing an important role in the design of the ultrasensitive methanol gas sensor. The sensing response and the optimum working temperature of ALSM and ALCM gas sensor are 52.29 and 155 °C and 34.89 and 155 °C, respectively, for 5 ppm methanol, and the highest response to other gases is 8. The ALSM and ALCM gas sensors reveal good selectivity and response for methanol.

Zn-responsive proteome profiling and time-dependent expression of proteins regulated by MTF-1 in A549 cells.

Zinc plays a critical role in many biological processes. However, it is toxic at high concentrations and its homeostasis is strictly regulated by metal-responsive transcription factor 1 (MTF-1) together with many other proteins to protect cells against metal toxicity and oxidative stresses. In this paper, we used high-resolution two-dimensional gel electrophoresis (2DE) to profile global changes of the whole soluble proteome in human lung adenocarcinoma (A549) cells in response to exogenous zinc treatment for 24 h. Eighteen differentially expressed proteins were identified by MALDI TOF/TOF and MASCOT search. In addition, we used Western blotting and RT-PCR to examine the time-dependent changes in expression of proteins regulated by MTF-1 in response to Zn treatment, including the metal binding protein MT-1, the zinc efflux protein ZnT-1, and the zinc influx regulator ZIP-1. The results indicated that variations in their mRNA and protein levels were consistent with their functions in maintaining the homeostasis of zinc. However, the accumulation of ZIP-1 transcripts was down-regulated while the protein level was up-regulated during the same time period. This may be due to the complex regulatory mechanism of ZIP-1, which is involved in multiple signaling pathways. Maximal changes in protein abundance were observed at 10 h following Zn treatment, but only slight changes in protein or mRNA levels were observed at 24 h, which was the time-point frequently used for 2DE analyses. Therefore, further study of the time-dependent Zn-response of A549 cells would help to understand the dynamic nature of the cellular response to Zn stress. Our findings provide the basis for further study into zinc-regulated cellular signaling pathways.

Effects of exogenous zinc on cell cycle, apoptosis and viability of MDAMB231, HepG2 and 293 T cells.

As a non-toxic metal to humans, zinc is essential for cell proliferation, differentiation, regulation of DNA synthesis, genomic stability and mitosis. Zinc homeostasis in cells, which is crucial for normal cellular functioning, is maintained by various protein families including ZnT (zinc transporter/SLC30A) and ZIP (Zrt-, Irt-like proteins/SLC39A) that decrease and increase cytosolic zinc availability, respectively. In this study, we investigated the influences of a specific concentration range of ZnSO4 on cell cycle and apoptosis by flow cytometry, and cell viability by MTT method in MDAMB231, HepG2 and 293 T cell lines. Fluorescent sensors NBD-TPEA and the counterstain for nuclei Hoechst 33342 were used to stain the treated cells for observing the localisation and amount of Zn(2+) via laser scanning confocal microscope. It was found that the influence manners of ZnSO4 on cell cycle, apoptosis and cell viability in various cell lines were different and corresponding to the changes of Zn(2+) content of the three cell lines, respectively. The significant increase on intracelluar zinc content of MDAMB231 cells resulted in cell death, G1 and G2/M cell cycle arrest and increased apoptotic fraction. Additionally, the mRNA expression levels of ZnT and ZIP families in the three cell lines, when treated with high concentration of ZnSO4, increased and decreased corresponding to their functions, respectively.

Effects of exogenous zinc on the cellular zinc distribution and cell cycle of A549 cells.

As the second most abundant transition metal in humans, zinc plays essential roles in normal cellular biological functions, including metabolism, signalling, proliferation, gene expression and apoptosis. We use ZnSO(4) as a stressor in this study to investigate for the first time the effects of exogenous Zn(2+) on both the cellular distribution of zinc and zinc-related proteins and the cell cycle of human lung adenocarcinoma (A549) cells. The cellular distribution of zinc and soluble proteins was determined in the whole cell as well as in the cytoplasmic and nuclear fractions. Exogenous zinc in the tested exposure range (0-100 µM) resulted in an altered cellular distribution of both zinc and the soluble proteins, together with total glutathione (GSx), the ratio of glutathione (GSH) to glutathione disulfide (GSSG) and non-protein sulphydryl (NPSH). Surprisingly, a turning point was observed in the re-distribution trend at a concentration of approximately 50 µM ZnSO(4). It is concluded that there exists a regulatory system in A549 cells that maintains the cellular zinc content stable in the presence of a certain range of extracellular zinc concentration. In addition, an MTT assay and flow cytometric analysis showed that the ZnSO(4) treatment led to a bi-phasic variation in viability and a slight fluctuation in the apoptosis of A549 cells. Our results will help to further elucidate zinc-related cell biology and biochemistry.

Clozapine v. chlorpromazine in treatment-naive, first-episode schizophrenia: 9-year outcomes of a randomised clinical trial.

BACKGROUND: The differential effects of so-called 'first- and second generation' antipsychotic medications, when given in the first episode, on the long-term outcome of schizophrenia remain to be elucidated. AIMS: We compared the 9-year outcomes of individuals initially randomised to clozapine or chlorpromazine. METHOD: One-hundred and sixty individuals with treatment-naive, first episode schizophrenia or schizophreniform disorder in a mental health centre in Beijing, China were randomised to clozapine or chlorpromazine treatment for up to 2 years,followed by up to an additional 7 years of naturalistic treatment. The primary outcome was remission status for individuals in each group. RESULTS: Individuals in both groups spent essentially equal amounts of time in each clinical state over the follow-up time period(remission, 78%; intermediate, 8%; relapse, 14%). There were no significant differences on other measures of illness severity. The clozapine group was more likely than the chlorpromazine group to remain on the medication to which they were originally assigned (26% v. 10%, P = 0.01). There were no significant differences between the two groups on other secondary efficacy outcomes. CONCLUSIONS: These findings support the comparability in effectiveness between antipsychotic medications but with slightly greater tolerability of clozapine in the treatment of first-episode psychosis.