Clcn3 deficiency ameliorates high-fat diet-induced obesity and improves metabolism in mice.

Objective: Obesity is defined as excess body fat and is a current health epidemic associated with increased risk for type 2 diabetes and cardiovascular disease. The ClC-3 chloride channel/antiporter, encoded by the Clcn3, is associated with some diseases, like carcinoma, nervous system diseases, and metabolic diseases. To verify the relationship between the Clcn3 and weight including metabolic changes, searching for a new target for metabolic therapy of obesity, we designed the experiment. Methods: The mice were divided into 4 different groups: Clcn3+/+ mice + high-fat diet (HFD), Clcn3-/- mice + HFD, Clcn3+/+ mice + normal diet (ND), Clcn3-/- mice + ND, and fed for 16 weeks. After the glucose tolerance test and insulin tolerance test, peripheral blood and adipose tissues were collected. Moreover, we performed transcriptome sequencing for the epididymal white adipose tissue from Clcn3+/+ and Clcn3-/- mice with the high-fat diet. Western blotting verified the changes in protein levels of relevant metabolic genes. Results: We found that the Clcn3-/- mice had lower body weight and visceral fat, refining glucose and lipid metabolism in HFD-induced mice, but had no effect in normal diet mice. RNA-seq and Western blotting indicated that Clcn3 deficiency may inhibit obesity through the AMPK-UCP1 axis. Conclusion: Modulation of Clcn3 may provide an appealing therapeutic target for obesity and associated metabolic syndrome.

Stochastic neuro-fuzzy system implemented in memristor crossbar arrays.

Neuro-symbolic artificial intelligence has garnered considerable attention amid increasing industry demands for high-performance neural networks that are interpretable and adaptable to previously unknown problem domains with minimal reconfiguration. However, implementing neuro-symbolic hardware is challenging due to the complexity in symbolic knowledge representation and calculation. We experimentally demonstrated a memristor-based neuro-fuzzy hardware based on TiN/TaOx/HfOx/TiN chips that is superior to its silicon-based counterpart in terms of throughput and energy efficiency by using array topological structure for knowledge representation and physical laws for computing. Intrinsic memristor variability is fully exploited to increase robustness in knowledge representation. A hybrid in situ training strategy is proposed for error minimizing in training. The hardware adapts easier to a previously unknown environment, achieving ~6.6 times faster convergence and ~6 times lower error than deep learning. The hardware energy efficiency is over two orders of magnitude greater than field-programmable gate arrays. This research greatly extends the capability of memristor-based neuromorphic computing systems in artificial intelligence.

LRRC8A as a central mediator promotes colon cancer metastasis by regulating PIP5K1B/PIP2 pathway.

Colorectal cancer (CRC) has been the third most common malignancy and the second cause of cancer-related mortality. As the core of volume-sensitive chloride currents, leucine-rich repeat-containing 8A (LRRC8A) contributes to tumor progression but is not consistent, especially for whom the roles in colon carcinoma metastasis were not fully elucidated. Herein, LRRC8A proteins were found highly expressed in hematogenous metastasis from human colorectal cancer samples. The oxaliplatin-resistant HCT116 cells highly expressed LRRC8A, which was related to impaired proliferation and enhanced migration. The over-expressed LRRC8A slowed proliferation and increased migration ex vivo and in vivo. The elevated LRRC8A upregulated the focal adhesion, MAPK, AMPK, and chemokine signaling pathways via phosphorylation and dephosphorylation. Inhibition of LRRC8A impeded the TNF-α signaling cascade and TNF-α-induced migration. LRRC8A binding to PIP5K1B regulated the PIP2 formation, providing a platform for LRRC8A to mediate cell signaling transduction. Importantly, LRRC8A self-regulated its transcription via NF-κB1 and NF-κB2 pathways and the upregulation of NIK/NF-κB2/LRRC8A transcriptional axis was unfavorable for colon cancer patients. Collectively, our findings reveal that LRRC8A is a central mediator in mediating multiple signaling pathways to promote metastasis and targeting LRRC8A proteins could become a potential clinical biomarker-driven treatment strategy for colon cancer patients.

Preliminary exploration of the correlation between spectral computed tomography quantitative parameters and spread through air spaces in lung adenocarcinoma.

Background: The invasive pattern called spread through air spaces (STAS) is linked to an unfavorable prognosis in patients with lung adenocarcinoma (LUAD). Using computed tomography (CT) signs alone to assess STAS is subjective and lacks quantitative evaluation, whereas spectral CT can provide quantitative analysis of tumors. The aim of this study was to investigate the association between spectral CT quantitative parameters and STAS in LUAD. Methods: We retrospectively collected consecutive patients with LUAD who underwent surgical resection and preoperative spectral CT scan at our institution. The quantitative parameters included CT values at 40, 70, and 100 keV [CT40keVa/v, CT70keVa/v, and CT100keVa/v (a: arterial; v: venous)]; iodine concentration (ICa/ICv); normalized iodine concentration (NICa/NICv); and slope λHU of the spectral curve (λHUa/λHUv). Clinical and CT features of the patients were also collected. Statistical analysis was performed to identify the quantitative parameters, clinical and CT features that were significantly correlated with STAS status. We evaluated the diagnostic performance of significant factors or models which combined quantitative parameters and CT features, using the area under the curve (AUC) of the receiver operating characteristic (ROC) curve. Results: We enrolled a total of 47 patients, with 32 positive and 15 negative for STAS. The results revealed that CT100keVa (P=0.002), CT100keVv (P=0.007), pathologic stage (P=0.040), tumor density (P<0.001), spiculation (P=0.003), maximum solid component diameter (P=0.008), and the consolidation/tumor ratio (CTR) (P=0.001) were significantly correlated with STAS status. The tumor density demonstrated a superior diagnostic capability [AUC =0.824, 95% confidence interval (CI): 0.709-0.939, sensitivity =59.4%, specificity =100.0%] compared to other variables. CT100keVa exhibited the best diagnostic performance (AUC =0.779, 95% CI: 0.633-0.925, sensitivity =78.1%, specificity =80.0%) among the quantitative parameters. Combination models were then constructed by combining the quantitative parameters with CT features. The total combined model showed the highest diagnostic efficiency (AUC =0.952, 95% CI: 0.894-1.000, sensitivity =90.6%, specificity =86.7%). Conclusions: Spectral CT quantitative parameters CT100keVa and CT100keVv may be potentially useful parameters in distinguishing the STAS status in LUAD.

Epigenetic and transcriptomic alterations in the ClC-3-deficient mice consuming a normal diet.

Introduction: Metabolic disorders are an important health concern that threatens life and burdens society severely. ClC-3 is a member of the chloride voltage-gated channel family, and ClC-3 deletion improved the phenotypes of dysglycemic metabolism and the impairment of insulin sensitivity. However, the effects of a healthy diet on transcriptome and epigenetics in ClC-3-/- mice were not explained in detail. Methods: Here, we performed transcriptome sequencing and Reduced Representation Bisulfite Sequencing for the liver of 3 weeks old WT and ClC-3-/- mice consuming a normal diet to insight into the epigenetic and transcriptomic alterations of ClC-3 deficient mice. Results: In the present study, we found that ClC-3-/- mice that were younger than 8 weeks old had smaller bodies compared to ClC-3+/+ mice with ad libitum self-feeding normal diet, and ClC-3-/- mice that were older than 10 weeks old had a similar body weight. Except for the spleen, lung, and kidney, the average weight of the heart, liver, and brain in ClC-3-/- mice was lower than that in ClC-3+/+ mice. TG, TC, HDL, and LDL in fasting ClC-3-/- mice were not significantly different from those in ClC-3+/+ mice. Fasting blood glucose in ClC-3-/- mice was lower than that in ClC-3+/+ mice; the glucose tolerance test indicated the response to blood glucose increasing for ClC-3-/- mice was torpid, but the efficiency of lowering blood glucose was much higher once started. Transcriptomic sequencing and reduced representation bisulfite sequencing for the liver of unweaned mice indicated that ClC-3 deletion significantly changed transcriptional expression and DNA methylation levels of glucose metabolism-related genes. A total of 92 genes were intersected between DEGs and DMRs-targeted genes, of which Nos3, Pik3r1, Socs1, and Acly were gathered in type II diabetes mellitus, insulin resistance, and metabolic pathways. Moreover, Pik3r1 and Acly expressions were obviously correlated with DNA methylation levels, not Nos3 and Socs1. However, the transcriptional levels of these four genes were not different between ClC-3-/- and ClC-3+/+ mice at the age of 12 weeks. Discussion: ClC-3 influenced the methylated modification to regulate glucose metabolism, of which the gene expressions could be driven to change again by a personalized diet-style intervention.

LRRC8A promotes the initial development of oxaliplatin resistance in colon cancer cells.

Leucine-rich repeat-containing 8 A (LRRC8A) is an essential component of the volume-regulated anion channel (VRAC), which plays a vital role in cell proliferation, migration, apoptosis, and drug resistance. In this study, we investigated the effects of LRRC8A on oxaliplatin resistance in colon cancer cells. The cell viability was measured after oxaliplatin treatment with cell counting kit-8 (CCK8) assay. RNA sequencing was used to analyze the differentially expressed genes (DEGs) between HCT116 and oxaliplatin-resistant HCT116 cell line (R-Oxa) cells. CCK8 assay and apoptosis assay indicated that R-Oxa cells significantly promoted drug resistance to oxaliplatin compared with native HCT116 cells. R-Oxa cells, deprived of oxaliplatin treatment for over six months (R-Oxadep), maintained a similar resistant property as R-Oxa cells. The LRRC8A mRNA and protein expression were markedly increased in both R-Oxa and R-Oxadep cells. Regulation of LRRC8A expression affected the resistance to oxaliplatin in native HCT116 cells, but not R-Oxa cells. Furthermore, The transcriptional regulation of genes in the platinum drug resistance pathway may contribute to the maintenance of oxaliplatin resistance in colon cancer cells. In conclusion, we propose that LRRC8A promotes the acquisition rather than the maintenance of oxaliplatin resistance in colon cancer cells.

LRRC8A is responsible for exosome biogenesis and volume regulation in colon cancer cells.

Exosomes are vital mediators for intercellular communications in the tumor microenvironment to accelerate colon cancer progression. Leucine-rich repeat-containing 8A (LRRC8A), the core component of the volume-regulated anion channel, is closely associated with acquiring heterogeneity for tumor cells. However, the role of LRRC8A in the exosomes remains largely unknown. Here, we reported that LRRC8A was one of the compositions in the exosomes released from colon cancer HCT116 cells. Down-regulation of LRRC8A proteins inhibited ex vivo cell growth and induced apoptosis. Consistently, chloride channel blockers DCPIB and NPPB inhibited cell growth and induced cell apoptosis in a time or concentration-dependent manner. Interestingly, the total amounts and proportions of different diameter exosomes released in 6 h were not altered by the treatment of DCPIB and NPPB in HCT116 cells. In contrast with the inhibition of LRRC8A, overexpression of LRRC8A proteins in HCT116 cells released significantly more distinct populations of exosomes. Importantly, the switches of ratios for exosomes in a hypotonic challenge were eliminated by DCPIB treatment. Collectively, our results uncovered that LRRC8A proteins were responsible for the exosome generation and sorted into exosomes for monitoring the volume regulation.

Effects of Shellfish and Organic Fertilizer Amendments on Soil Nutrients and Tea Yield and Quality.

Soil acidification in tea plantations leads to an excessive heavy metal content in tea, decreasing its yield and quality. How to apply shellfish and organic fertilizers to improve soil and ensure the safe production of tea is still not clear. A two-year field experiment was conducted in tea plantations in which the soil was characterized by a pH of 4.16 and concentrations of lead (Pb) (85.28 mg/kg) and cadmium (Cd) (0.43 mg/kg) exceeding the standard. We used shellfish amendments (750, 1500, 2250 kg/ha) and organic fertilizers (3750, 7500 kg/ha) to amend the soils. The experimental results showed that compared with the treatment without any amendment (CK), the soil pH increased by 0.46 on average; the soil available nitrogen, phosphorus, and potassium contents increased by 21.68%, 19.01%, and 17.51% respectively; and the soil available Pb, Cd, Cr, and As contents decreased by 24.64%, 24.36%, 20.83%, and 26.39%, respectively. In comparison to CK, the average yield of tea also increased by 90.94 kg/ha; tea polyphenols, free amino acids, caffeine, and water extract increased by 9.17%, 15.71%, 7.54%, and 5.27%, respectively; and the contents of Pb, Cd, As, and Cr in the tea decreased significantly (p < 0.05) by 29.44-61.38%, 21.43-61.38%, 10.43-25.22%, and 10.00-33.33%, respectively. The greatest effects on all parameters occurred with the largest amendment of both shellfish (2250 kg/ha) and organic fertilizer (7500 kg/ha) combined. This finding suggests that the optimized amendment of shellfish could be used as a technical measure to improve the health quality of both soil and tea in acidified tea plantations in the future.

Insights into Genomic Evolution and the Potential Genetic Basis of Klebsiella variicola subsp. variicola ZH07 Reveal Its Potential for Plant Growth Promotion and Autotoxin Degradation.

The accumulation of autotoxins in soil causes continuous cropping obstacle stress in crops, and the bioremediation of autotoxins by microorganisms is an efficient process. In this study, strain ZH07 was isolated from the peanut rhizosphere and was found to be utilizing multiple autotoxins as its carbon sources. Based on its genomic characteristics and a phylogenetic analysis, ZH07 represents a member of Klebsiella variicola subsp. variicola. A comparative genomic analysis exhibited evolutionary dynamics exhibited by mobile genetic elements (MGEs), strain-specific genes, potential horizontal genes, and evolutionary constraints driven by purifying selection, which facilitated its genomic adaptation to rhizosphere soil. Genome mining revealed the potential genomic properties associated with plant growth promotion, such as nitrogen fixation, indole acetic acid synthesis, phosphonate solubilization and assimilation, siderophore production, and secondary metabolite synthesis. Moreover, abundant genes putatively responsible for the biodegradation of aromatic xenobiotics, including benzoic acid, cinnamic acid, vanillic acid, protocatechuic acid, phenylacetic acid, and p-hydroxybenzoic acid were also observed in the ZH07 genome. Compared to autotoxin stress alone, the combination of ZH07 application promoted peanut germination and seedling growth. Our analysis revealed the genetic adaptation of ZH07 to the rhizosphere environment and the potential genetic basis and effectiveness of the isolate to serve as a plant growth stimulator. IMPORTANCE Continuous cropping obstacles reduce the production and quality of agricultural products, and the application of rhizosphere beneficial microbes is an important strategy. Strain ZH07 showed autotoxin-degrading and plant growth-promoting capacities. The objectives of this study were to characterize its genomic evolution and the potential genetic basis of the autotoxin degradation and plant growth promotion. ZH07 represents a member of Klebsiella variicola subsp. variicola, based on genomic and phylogenetic analyses. Its genomic components have undergone different degrees of purifying selection, and the disparity in the evolutionary rate may be associated with its niche adaptation. A systematic analysis of the ZH07 genome identified the potential genetic basis that contributes to plant growth promotion and to aromatic xenobiotic biodegradation. This study demonstrates that plant growth-promoting rhizobacteria (PGPR) play important roles in autotoxin biodegradation and can be used as biofertilizers to enhance the growth of peanuts in response to continuous cropping obstacle stress.

Pollution Characteristics, Sources, and Health Risk Assessments of Potentially Toxic Elements in Community Garden Soil of Lin'an, Zhejiang, China.

The characteristics, sources and risk assessment of heavy metal pollution in community garden soil of Lin'an District were evaluated. The 28 soil samples from community garden were collected for determination of 7 heavy metal elements. The Geostatistical analysis, Spearman correlation coefficient, Principal component analysis and PMF model have explored sources of heavy metal pollution. The health risk assessment model has assessed ecological risk of heavy metals. The results revealed that average concentration of As, Cd, Cr, Cu, Ni, Pb and Zn were 16.0, 0.158, 76.1, 34.6, 45.8, 20.9 and 166 mg kg-1, respectively. Whereas As, Cd, Cr, Cu, Ni and Zn were higher than background values. The spatial distribution of heavy metal pollution in the southwest of the study area was higher than northeast. The pollution sources of Cd, Cu, Ni and Zn in the study area were due to agricultural activities (42.9%), Cr and Pb were from traffic sources (36.2%), and As was domestic pollution (20.9%) according to Spearman correlation coefficient, Principal component analysis and PMF model. The non-carcinogenic risks of As (5.39), Cr (3.53) and Ni (2.07) have a value of 1, which indicated significant risk. The potentially toxic elements have not exceeded maximum threshold of USEPA, with regard to carcinogenic risk, while As (3.37E-05) and Cr (5.74E-05) have exceeded the safety range. It is concluded that soils of community gardens are facing pollution problem due to potentially toxic elements which require environmental monitoring of the soil to reduce risk of human health.

Insights into Evolutionary, Genomic, and Biogeographic Characterizations of Chryseobacterium nepalense Represented by a Polyvinyl Alcohol-Degrading Bacterium, AC3.

Chryseobacterium spp. are Gram-negative rods found ubiquitously in the environment, with certain species being reported as having unusual degrading properties. Polyvinyl alcohol (PVA) is used widely in industry but causes serious global environmental pollution. Here, we report the complete genome sequence of a novel bacterium, AC3, that efficiently degrades PVA. As the representative genome of Chryseobacterium nepalense, key genomic characteristics (e.g., mobile genetic elements, horizontal genes, genome-scale metabolic network, secondary metabolite biosynthesis gene clusters, and carbohydrate-active enzymes) were comprehensively investigated to reveal the potential genetic features of this species. Core genome phylogenetic analysis in combination with average nucleotide identity, average amino acid identity, and in silico DNA-DNA hybridization values provided an accurate taxonomic position of C. nepalense in the genus Chryseobacterium. Comparative genomic analysis of AC3 with closely related species suggested evolutionary dynamics characterized by a species-specific genetic repertoire, dramatic rearrangements, and evolutionary constraints driven by selective pressure, which facilitated the speciation and adaptative evolution of C. nepalense. Biogeographic characterization indicated that this species is ubiquitously distributed not only in soil habitats but also in a variety of other source niches. Bioinformatic analysis revealed the potential genetic basis of PVA degradation in AC3, which included six putative genes associated with the synthesis of PVA dehydrogenase, cytochrome c, oxidized PVA hydrolase, and secondary alcohol dehydrogenase. Our study reports the first complete genome of C. nepalense with PVA-degrading properties, providing comprehensive insights into the genomic characteristics of this species and increasing our understanding of the microbial degradation of PVA. IMPORTANCE Although PVA is a biodegradable polymer, the widespread use of PVA in global industrialization has resulted in serious environmental problems. To date, knowledge of effective and applicable PVA-degrading bacteria is limited, and thus, the discovery of novel PVA biodegraders is pertinent. Here, we isolated a novel bacterial strain, AC3, which efficiently degraded PVA. The complete genome of AC3 was sequenced as the first genome sequence of the species C. nepalense. Comparative genomic analysis was performed to comprehensively investigate the phylogenetic relationships, genome-scale metabolic network, key genomic characteristics associated with genomic evolution, evolutionary dynamics between AC3 and its close relatives, and biogeographic characterization of C. nepalense, particularly regarding the potential genetic basis of PVA degradation. These findings could advance our understanding of the genomic characteristics of C. nepalense and PVA bioremediation.

Pan-Genome Analysis of Delftia tsuruhatensis Reveals Important Traits Concerning the Genetic Diversity, Pathogenicity, and Biotechnological Properties of the Species.

Delftia tsuruhatensis strains have long been known to promote plant growth and biological control. Recently, it has become an emerging opportunistic pathogen in humans. However, the genomic characteristics of the genetic diversity, pathogenicity, and biotechnological properties have not yet been comprehensively investigated. Here, a comparative pan-genome analysis was constructed. The open pan-genome with a large and flexible gene repertoire exhibited a high degree of genetic diversity. The purifying selection was the main force to drive pan-genome evolution. Significant differences were observed in the evolutionary relationship, functional enrichment, and degree of selective pressure between the different components of the pan-genome. A high degree of genetic plasticity was characterized by the determinations of diverse mobile genetic elements (MGEs), massive genomic rearrangement, and horizontal genes. Horizontal gene transfer (HGT) plays an important role in the genetic diversity of this bacterium and the formation of genomic traits. Our results revealed the occurrence of diverse virulence-related elements associated with macromolecular secretion systems, virulence factors associated with multiple nosocomial infections, and antimicrobial resistance, indicating the pathogenic potential. Lateral flagellum, T1SS, T2SS, T6SS, Tad pilus, type IV pilus, and a part of virulence-related genes exhibited general properties, whereas polar flagellum, T4SS, a part of virulence-related genes, and resistance genes presented heterogeneous properties. The pan-genome also harbors abundant genetic traits related to secondary metabolism, carbohydrate active enzymes (CAZymes), and phosphate transporter, indicating rhizosphere adaptation, plant growth promotion, and great potential uses in agriculture and biological control. This study provides comprehensive insights into this uncommon species from the genomic perspective. IMPORTANCE D. tsuruhatensis is considered a plant growth-promoting rhizobacterium (PGPR), an organic pollutant degradation strain, and an emerging opportunistic pathogen to the human. However, the genetic diversity, the evolutionary dynamics, and the genetic basis of these remarkable traits are still little known. We constructed a pan-genome analysis for D. tsuruhatensis and revealed extensive genetic diversity and genetic plasticity exhibited by open pan-genome, diverse mobile genetic elements (MGEs), genomic rearrangement, and horizontal genes. Our results highlight that horizontal gene transfer (HGT) and purifying selection are important forces in D. tsuruhatensis genetic evolution. The abundant virulence-related elements associated with macromolecular secretion systems, virulence factors, and antimicrobial resistance could contribute to the pathogenicity of this bacterium. Therefore, clinical microbiologists need to be aware of D. tsuruhatensis as an opportunistic pathogen. The genetic profiles of secondary metabolism, carbohydrate active enzymes (CAZymes), and phosphate transporter could provide insight into the genetic armory of potential applications for agriculture and biological control of D. tsuruhatensis in general.

Effects of palmitic acid on localization of embryo cell fate and blastocyst formation gene products.

As obese and overweight patients commonly display hyperlipidemia and are increasingly accessing fertility clinics for their conception needs, our studies are directed at understanding the effects of hyperlipidemia on early pregnancy. We have focused on investigating palmitic acid (PA) and oleic acid (OA) treatment alone and in combination from the mouse two-cell stage embryos as a model for understanding their effects on the mammalian preimplantation embryo. We recently reported that PA exerts a negative effect on mouse two-cell progression to the blastocyst stage, whereas OA co-treatment reverses that negative effect. In the present study, we hypothesized that PA treatment of mouse embryos would disrupt proper localization of cell fate determining and blastocyst formation gene products and that co-treatment with OA would reverse these effects. Our results demonstrate that PA treatment significantly (P < 0.05) reduces blastocyst development and cell number but did not prevent nuclear localization of YAP in outer cells. PA treatment significantly reduced the number of OCT4+ and CDX2+ nuclei. PA-treated embryos had lower expression of blastocyst formation proteins (E-cadherin, ZO-1 and Na/K-ATPase alpha1 subunit). Importantly, co-treatment of embryos with OA reversed PA-induced effects on blastocyst development and increased inner cell mass (ICM) and trophectoderm (TE) cell numbers and expression of blastocyst formation proteins. Our findings demonstrate that PA treatment does not impede cell fate gene localization but does disrupt proper blastocyst formation gene localization during mouse preimplantation development. OA treatment is protective and reverses PA's detrimental effects. The results advance our understanding of the impact of FFA exposure on mammalian preimplantation development.

Photo-induced bleaching and thermally stimulated recovery of BAC-P in Bi-doped phosphosilicate fibers.

The first results of the study on photobleaching and thermally induced recovery in Bi-doped phosphosilicate fiber have been presented. It was revealed that the rate of bleaching of phosphor-related Bi active center (BAC-P) becomes slower with the decrease of photon energy. The quadratic dependence of the bleaching rate of BAC-P on laser power is obtained under 532 nm laser irradiation. The effect of temperature on the bleaching dynamics of BAC-P is also investigated under 532 nm radiation, suggesting a thermally aggravated bleaching process upon heating at certain temperatures (≥300∘C). Furthermore, the thermal recovery of bleached Bi-doped silica-based fiber (BDF) is investigated and a 13% increase of luminescence is achieved upon thermal quenching for 5 min at 400ºC. The underlying mechanism of photobleaching and thermo-stimulated recovery process of BAC-P is also discussed.