Selenocompound Metabolism Research Articles

IN-VITRO EFFECTS OF HESPERIDIN ON HUMAN ERYTHROCYTES SICKLING WITH MODIFICATIONS OF FUNCTIONAL CHEMISTRY

Sickle cell disease (SCD) is still one of the highest leading inbred hemoglobinopathy amid Africans. The In-vitro effects of hesperidin on sickling human erythrocytes with functional chemistry of human sickle erythrocytes were investigated by standard methods.The induction of sickle cell in erythrocytes was done using 2%sodium metabisulphite at 3 hours optimally. The prevention and curative approaches (PA and CA) were adopted in comparison of human erythrocytesfrom AA-genotype. The erythrocytes were grouped into 3; AA–hesperidin treated (AA××), AA–hesperidin treated (AA×) and sickle cell serving as negative control (NC). The results showed 83% and 86% antisickling potency of hesperidin for prevention and curative approaches respectively. The results of functional chemistry analysis showed that hesperidin modifies the functional groups in favor of its antisickling effects judging from the FTIR observed bends and shifts. From GC-MS and LC-MS analyses, it was practical that hesperidin treatment favors the productions of fatty acid alkyl monoesters (FAMEs) with concomitant shutting down-effects on selenocompound metabolism. Pathway enrichment and topology analyses revealed non-specifically activation of fatty acid biosynthesis, linoleic acid metabolism and steroid hormone biosynthesis pathways upon hesperidin treatment. Thus, sickling-suppressive effects of hesperidin could potentially be associated with modulation of oxygenated and deoxygenated haemglobinthroughpossible modification of human sickle erythrocyte’s functional chemistry.

Identification of aberrantly methylated differentially expressed genes and pro-tumorigenic role of KIF2C in melanoma.

Background: Skin Cutaneous Melanoma (SKCM) is known as an aggressive malignant cancer, which could be directly derived from melanocytic nevi. However, the molecular mechanisms underlying the malignant transformation of melanocytes and melanoma tumor progression still remain unclear. Increasing research showed significant roles of epigenetic modifications, especially DNA methylation, in melanoma. This study focused on the identification and analysis of methylation-regulated differentially expressed genes (MeDEGs) between melanocytic nevus and malignant melanoma in genome-wide profiles. Methods: The gene expression profiling datasets (GSE3189 and GSE114445) and gene methylation profiling datasets (GSE86355 and GSE120878) were downloaded from the Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) and differentially methylated genes (DMGs) were identified via GEO2R. MeDEGs were obtained by integrating the DEGs and DMGs. Then, a functional enrichment analysis of MeDEGs was performed. STRING and Cytoscape were used to describe the protein-protein interaction (PPI) network. Furthermore, survival analysis was implemented to select the prognostic hub genes. Next, we conducted gene set enrichment analysis (GSEA) of hub genes. To validate, SKCM cell culture and lentivirus infection was performed to reveal the expression and behavior pattern of KIF2C. Patients and specimens were collected and then immunohistochemistry (IHC) staining was conducted. Results: We identified 237 hypomethylated, upregulated genes and 182 hypermethylated, downregulated genes. Hypomethylation-upregulated genes were enriched in biological processes of the oxidation-reduction process, cell proliferation, cell division, phosphorylation, extracellular matrix disassembly and protein sumoylation. Pathway enrichment showed selenocompound metabolism, small cell lung cancer and lysosome. Hypermethylation-downregulated genes were enriched in biological processes of positive regulation of transcription from RNA polymerase II promoter, cell adhesion, cell proliferation, positive regulation of transcription, DNA-templated and angiogenesis. The most significantly enriched pathways involved the transcriptional misregulation in cancer, circadian rhythm, tight junction, protein digestion and absorption and Hippo signaling pathway. After PPI establishment and survival analysis, seven prognostic hub genes were CKS2, DTL, KIF2C, KPNA2, MYBL2, TPX2, and FBL. Moreover, the most involved hallmarks obtained by GSEA were E2F targets, G2M checkpoint and mitotic spindle. Importantly, among the 7 hub genes, we found that down-regulated level of KIF2C expression significantly inhibited the proliferative ability of SKCM cells and suppressed the metastasis capacity of SKCM cells. Conclusions: Our study identified potential aberrantly methylated-differentially expressed genes participating in the process of malignant transformation from nevus to melanoma tissues based on comprehensive genomic profiles. Transcription profiles of CKS2, DTL, KIF2C, KPNA2, MYBL2, TPX2, and FBL provided clues of aberrantly methylation-based biomarkers, which might improve the development of precision medicine. KIF2C plays a pro-tumorigenic role and potentially inhibited the proliferative ability in SKCM.

RNA-seq analysis of green tea polyphenols modulation of differently expressed genes in Enterococcus faecalis under low pH.

Enterococcus faecasslis (E. faecalis) is a resident bacterium in the host. The increase in internal stress like low pH may affect the biological effects of E. faecalis. The prebiotic-like function of tea polyphenols can enhance the beneficial effects of its tolerance to environmental stress. In this study, RNA-sequence analysis was used to explore the protective effect of green tea polyphenols (GTP) on E. faecalis under low pH stress. A total of 28 genes were found to be responsive to GTP under low pH stress, including 16 up-regulated and 12 down-regulated. GTP intervention can partly relieve some undesired negative influences, such as the down-regulation of the base excision repair gene and amino acid transport and metabolism gene. The significantly changes were associated with selenocompound metabolism and aminoacyl-tRNA biosynthesis after the intervention of GTP. The present study provided new insights into the growth and continuous adaptation of E. faecalis under stress.

Distinct gut microbiota and health outcomes in asymptomatic infection, viral nucleic acid test re-positive, and convalescent COVID-19 cases.

Gut microbiota composition is suggested to associate with coronavirus disease 2019 (COVID‐19) severity, but the impact of gut microbiota on health outcomes is largely unclear. We recruited 81 individuals from Wuhan, China, including 13 asymptomatic infection cases (Group A), 24 COVID‐19 convalescents with adverse outcomes (Group C), 31 severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) re‐positive cases (Group D), and 13 non‐COVID‐19 healthy controls (Group H). The microbial features of Groups A and D were similar and exhibited higher gut microbial diversity and more abundant short‐chain fatty acid (SCFA)‐producing species than Group C. Group C was enriched with opportunistic pathogens and virulence factors related to adhesion and toxin production. The abundance of SCFA‐producing species was negatively correlated, while Escherichia coli was positively correlated with adverse outcomes. All three groups (A, C, and D) were enriched with the mucus‐degrading species Akkermansia muciniphila, but decreased with Bacteroides‐encoded carbohydrate‐active enzymes. The pathways of vitamin B6 metabolic and folate biosynthesis were decreased, while selenocompound metabolism was increased in the three groups. Specifically, the secondary bile acid (BA) metabolic pathway was enriched in Group A. Antibiotic resistance genes were common among the three groups. Conclusively, the gut microbiota was related to the health outcomes of COVID‐19. Dietary supplementations (SCFAs, BA, selenium, folate, vitamin B6) may be beneficial to COVID‐19 patients.

The transcriptomic analysis revealed sulfamethoxazole stress at environmentally relevant concentration on the mechanisms of toxicity of cyanobacteria Synechococcus sp

The study addressed the mechanisms of toxicity induced to cyanobacteria Synechococcus sp by its exposure to sulfamethoxazole (SMX). Here, transcriptomics analysis was used to investigate variations in the molecular level of Synechococcus sp under SMX stress at concentrations of 0.5 and 10 μg L−1. There were insignificant differences in algal density between 0.5 μg L−1 and the control group, whereas a conspicuous reduction of 56% in the 10 μg L−1 group was discovered. Additionally, 12 and 133 differentially expressed genes (DEGs) were found in the different treatment groups. Experimental results show that SMX at the environmentally realistic concentration may not disrupt the algal physiological status. Cyanobacteria are similar to bacterial in structure, and SMX has been recognized antibacterial effect of inhibiting folate synthesis. However, DEGs were dominantly enriched in the porphyrin and chlorophyll metabolism, photosynthesis, selenocompound metabolism, and tocopherol biosynthesis rather than folate biosynthesis pathway, indicating SMX has different mode of action between Synechococcus sp and bacteria. briefly, the Synechococcus sp was used as model organisms to reveal the mechanism of SMX.

Untargeted serum metabolomics and potential biomarkers for Sjögren's syndrome.

At present, the pathogenesis of Sjögren's syndrome (SS) remains unclear. This research aimed to identify differential metabolites that contribute to SS diagnosis and discover the disturbed metabolic pathways. Recent advances in mass spectrometry have allowed the identification of hundreds of unique metabolic signatures and the exploration of altered metabolite profiles in disease. In this study, 505 candidates including healthy controls (HCs) and SS patients were recruited and the serum samples were collected. A non-targeted gas chromatography-mass spectrometry (GC-MS) serum metabolomics method was used to explore the changes in serum metabolites. We found SS patients and HCs can be distinguished by 21 significant metabolites. The levels of alanine, tryptophan, glycolic acid, pelargonic acid, cis-1-2-dihydro-1-2-naphthalenediol, diglycerol, capric acid, turanose, behenic acid, dehydroabietic acid, stearic acid, linoleic acid, heptadecanoic acid, valine, and lactic acid were increased in serum samples from SS patients, whereas levels of catechol, anabasine, 3-6-anhydro-D-galactose, beta-gentiobiose, 2-ketoisocaproic acid and ethanolamine were decreased. The significantly changed pathways included the following: Linoleic acid metabolism; unsaturated fatty acid biosynthesis; aminoacyl-tRNA biosynthesis; valine, leucine, and isoleucine biosynthesis; glycerolipid metabolism; selenocompound metabolism; galactose metabolism; alanine, aspartate and glutamate metabolism; glyoxylate and dicarboxylate metabolism; glycerophospholipid metabolism; and valine, leucine and isoleucine degradation. These findings enhance the informative capacity of biochemical analyses through the identification of serum biomarkers and the analysis of metabolic pathways and contribute to an improved understanding of the pathogenesis of SS.

Cluster analysis and profiling of airway fluid metabolites in pediatric acute hypoxemic respiratory failure

Hierarchal clustering of amino acid metabolites may identify a metabolic signature in children with pediatric acute hypoxemic respiratory failure. Seventy-four immunocompetent children, 41 (55.4%) with pediatric acute respiratory distress syndrome (PARDS), who were between 2 days to 18 years of age and within 72 h of intubation for acute hypoxemic respiratory failure, were enrolled. We used hierarchal clustering and partial least squares-discriminant analysis to profile the tracheal aspirate airway fluid using quantitative LC–MS/MS to explore clusters of metabolites that correlated with acute hypoxemia severity and ventilator-free days. Three clusters of children that differed by severity of hypoxemia and ventilator-free days were identified. Quantitative pathway enrichment analysis showed that cysteine and methionine metabolism, selenocompound metabolism, glycine, serine and threonine metabolism, arginine biosynthesis, and valine, leucine, and isoleucine biosynthesis were the top five enriched, impactful pathways. We identified three clusters of amino acid metabolites found in the airway fluid of intubated children important to acute hypoxemia severity that correlated with ventilator-free days < 21 days. Further studies are needed to validate our findings and to test our models.

Is selenium beneficial or detrimental to earthworm? Growth and metabolism responses of Eisenia Fetida to Na2SeO3 exposure

Se unevenly distributed in soils due to variations of geology and anthropogenic input, which results in different effects on earthworms. The effects of Se were characterized by analyzing the growth and metabolism responses of earthworms after exposure to three different concentrations of Na2SeO3. The results showed that except the possible growth promotion at 5 mg/kg, low and middle-level exposure to Na2SeO3 (0.3–10 mg/kg) did not significantly affect the growth of earthworms. While a significant inhibition effect on growth was observed in the high-level exposure group (30–70 mg/kg). There was an inflection point for Se performing promotion to inhibition effects on earthworm growth. To investigate the metabolic response of earthworms, a novel HPLC-ESI-MS (High Performance Liquid Chromatography-Electrospray Ionization-Mass Spectrometry) method was used to determine sensitive biomarkers. Selenium exposure significantly altered the metabolism of seven essential amino acids, namely tyrosine, leucine, phenylalanine, valine, alanine, glycine, and lysine, and two selenoamino acids, namely selenomethionine and methylselenocysteine. The overall metabolism level of earthworms was not affected at low exposure concentrations, but was affected at medium and high exposure concentrations. The metabolic pathways that integrated the selenocompound metabolism and the tricarboxylic acid cycle from the perspective of energy supply and demand were affected by Na2SeO3 exposure. The derived reactive oxygen species at high exposure concentrations were probably the reason for the growth inhibition effect of Se on earthworms. This study provides biochemical insights into the effects of Na2SeO3 on earthworms and suggests that an Se concentration of about 2.3 mg/kg is appropriate for soil organism health.

The Cardamine enshiensis genome reveals whole genome duplication and insight into selenium hyperaccumulation and tolerance

Cardamine enshiensis is a well-known selenium (Se)-hyperaccumulating plant. Se is an essential trace element associated with many health benefits. Despite its critical importance, genomic information of this species is limited. Here, we report a chromosome-level genome assembly of C. enshiensis, which consists of 443.4 Mb in 16 chromosomes with a scaffold N50 of 24 Mb. To elucidate the mechanism of Se tolerance and hyperaccumulation in C. enshiensis, we generated and analyzed a dataset encompassing genomes, transcriptomes, and metabolomes. The results reveal that flavonoid, glutathione, and lignin biosynthetic pathways may play important roles in protecting C. enshiensis from stress induced by Se. Hi-C analysis of chromatin interaction patterns showed that the chromatin of C. enshiensis is partitioned into A and B compartments, and strong interactions between the two telomeres of each chromosome were correlated with histone modifications, epigenetic markers, DNA methylation, and RNA abundance. Se supplementation could affect the 3D chromatin architecture of C. enshiensis at the compartment level. Genes with compartment changes after Se treatment were involved in selenocompound metabolism, and genes in regions with topologically associated domain insulation participated in cellular responses to Se, Se binding, and flavonoid biosynthesis. This multiomics research provides molecular insight into the mechanism underlying Se tolerance and hyperaccumulation in C. enshiensis.

Molecular Insights into Beneficial Effects of Tea-Plant Growth and Selenium Enrichment by Herbaspirillum camelliae

Herbaspirillum camelliae WT00C, as a tea-plant endophytic bacterium, not only colonizes specifically in tea plants but also promotes tea-plant growth and selenium enrichment. Different from diazotrophic endophytes H. seropedicae, H. frisingense and H. rubrisubalbicans, H. camelliae WT00C does not display nitrogen-fixing activity. To understand the molecular mechanisms of promoting the growth of tea plant and Se-enrichment, we sequenced and annotated the genome of H. camelliae WT00C. The results showed that the genome was composed of 6,079,821 base pairs with a total of 5,537 genes. The genomic survey also revealed that H. camelliae WT00C was a multifunctional bacterium metabolizing a variety of carbon and nitrogen sources and defending against biotic and abiotic stress. Although this bacterium did not have intact nitrogen-fixing genes, its genome held the genes responsible for indole-3-acetic acid (IAA) biosynthesis, 1-aminocyclopropane-1-carboxylate (ACC) deamination, siderophore synthesis, ammonia formation, urea metabolism, glutathione and selenocompound metabolisms. Biosynthesis of IAA, siderophore, ammonia, urea and ACC deaminase could explain why two bacterial strains promote tea-plant growth and development. Selenocompound metabolism in this bacterium might also benefit tea-plant growth and Se-enrichment. In addition, the genome of H. camelliae also contained a multitude of protein secretion systems T1SS, T3SS, T4SS and T6SS, in which T4SS did not exhibit in other members of the genus Herbaspirillum.

Antisickling effect of chrysin is associated with modulation of oxygenated and deoxygenated haemoglobin via alteration of functional chemistry and metabolic pathways of human sickle erythrocytes.

Sickle cell disease (SCD) treatment and management remain a challenging puzzle especially among developing Nations. Chrysin's sickling-suppressive properties in human sickle (SS) erythrocytes in addition to its effect on AA-genotype erythrocytes were evaluated. Sickling was induced (76%) with 2% sodium metabisulphite at 3 h. Chrysin prevented (81.19%) the sickling and reversed same (84.63%) with strong IC50s (0.0257 µM and 0.00275 µM, respectively). The levels of oxygenated haemoglobin in the two groups (before and after induction approaches) were similar but significantly (P < 0.05) higher than that of SS erythrocytes (the 'induced' control), with chrysin-treated AA-genotype showing no effects relative to the untreated. The level of deoxygenated haemoglobin in the 'induced' control group was significantly (P < 0.05) higher than those of the chrysin-treated SS erythrocytes. Normal and chrysin-untreated erythrocytes (AA-untreated) were significantly more resistant to osmotic fragility than the SS-untreated. However, treatment with chrysin significantly reduced the osmotic fragility of the cells relative to the untreated cells. Furthermore, chrysin treatment significantly lowers the high level of 2,3-diphosphoglycerate (2,3-DPG) observed in the sickle erythrocytes, with no effects on AA-genotype erythrocytes. Based on functional chemistry, chrysin treatment alters the functional groups in favour of its antisickling effects judging from the observed bends and shifts. From metabolomics analysis, it was observed that chrysin treatment favors fatty acid alkyl monoesters (FAMEs) production with concomitant shutting down-effects on selenocompound metabolism. Thus, sickling-suppressive effects of chrysin could potentially be associated with modulation of oxygenated and deoxygenated haemoglobin via alteration of human sickle erythrocyte's functional chemistry and metabolic pathways implicated in SCD crisis.

Comparison of the rabbit and human corneal endothelial proteomes regarding proliferative capacity

The shortage of human donor corneas has raised important concerns about engineering of corneal endothelial cells (CECs) for clinical use. However, due to the limited proliferative capacity of human CECs, driving them into proliferation and regeneration may be difficult. Unlike human CECs, rabbit CECs have a marked proliferative capacity. To clarify the potential reason for this difference, we analysed the proteomes of four human corneal endothelium samples and four rabbit corneal endothelium samples with quantitative label-free proteomics and downstream analysis. We discovered that vitamin and selenocompound metabolism and some signaling pathways such as NF-kappa B signaling pathway differed between the samples. Moreover, TGFβ, PITX2 and keratocan were distinctively expressed in rabbit samples, which might be associated with active proliferation in rabbit CECs. This study illustrates the proteomic differences between human and rabbit CECs and might promote CEC engineering strategies.

Screening and Genetic Network Analysis of Genes Involved in Freezing and Thawing Resistance in DaMDHAR-Expressing Saccharomyces cerevisiae Using Gene Expression Profiling.

The cryoprotection of cell activity is a key determinant in frozen-dough technology. Although several factors that contribute to freezing tolerance have been reported, the mechanism underlying the manner in which yeast cells respond to freezing and thawing (FT) stress is not well established. Therefore, the present study demonstrated the relationship between DaMDHAR encoding monodehydroascorbate reductase from Antarctic hairgrass Deschampsia antarctica and stress tolerance to repeated FT cycles (FT2) in transgenic yeast Saccharomyces cerevisiae. DaMDHAR-expressing yeast (DM) cells identified by immunoblotting analysis showed high tolerance to FT stress conditions, thereby causing lower damage for yeast cells than wild-type (WT) cells with empty vector alone. To detect FT2 tolerance-associated genes, 3′-quant RNA sequencing was employed using mRNA isolated from DM and WT cells exposed to FT (FT2) conditions. Approximately 332 genes showed ≥2-fold changes in DM cells and were classified into various groups according to their gene expression. The expressions of the changed genes were further confirmed using western blot analysis and biochemical assay. The upregulated expression of 197 genes was associated with pentose phosphate pathway, NADP metabolic process, metal ion homeostasis, sulfate assimilation, β-alanine metabolism, glycerol synthesis, and integral component of mitochondrial and plasma membrane (PM) in DM cells under FT2 stress, whereas the expression of the remaining 135 genes was partially related to protein processing, selenocompound metabolism, cell cycle arrest, oxidative phosphorylation, and α-glucoside transport under the same condition. With regard to transcription factors in DM cells, MSN4 and CIN5 were activated, but MSN2 and MGA1 were not. Regarding antioxidant systems and protein kinases in DM cells under FT stress, CTT1, GTO, GEX1, and YOL024W were upregulated, whereas AIF1, COX2, and TRX3 were not. Gene activation represented by transcription factors and enzymatic antioxidants appears to be associated with FT2-stress tolerance in transgenic yeast cells. RCK1, MET14, and SIP18, but not YPK2, have been known to be involved in the protein kinase-mediated signalling pathway and glycogen synthesis. Moreover, SPI18 and HSP12 encoding hydrophilin in the PM were detected. Therefore, it was concluded that the genetic network via the change of gene expression levels of multiple genes contributing to the stabilization and functionality of the mitochondria and PM, not of a single gene, might be the crucial determinant for FT tolerance in DaMDAHR-expressing transgenic yeast. These findings provide a foundation for elucidating the DaMDHAR-dependent molecular mechanism of the complex functional resistance in the cellular response to FT stress.

Physicochemical and transcriptomic responses of Lactobacillus brevis JLD715 to sodium selenite.

Elemental selenium, as a new type of selenium supplement, can be prepared by microorganisms reducing inorganic selenium. In this study, Lactobacillus brevis JLD715 was incubated in broth containing different concentrations of sodium selenite (Na2 SeO3 ). The results showed that the bacterial biomass of L. brevis JLD715 decreased due to the inhibition of Na2 SeO3 . The cell membrane of L. brevis JLD715 treated with Na2 SeO3 was damaged, as evidenced by the reduction of intracellular ATP concentration, depolarization of cell membrane, reduction of intracellular pH and impairment of membrane integrity. In addition, we investigated the metabolism mechanism of Na2 SeO3 by L. brevis JLD715 based on transcriptome sequencing. A total of 461 genes were significantly differentially expressed under Na2 SeO3 treatment, of which 231 genes were up-regulated and 230 genes were down-regulated. These genes were involved in pathways such as pyruvate metabolism, fatty acid biosynthesis, selenocompound metabolism and nucleotide-binding oligomerization domain-like (NOD-like) receptor signaling. Meanwhile, the genes related to sulfhydryl oxidoreductase, electron carrier proteins and transmembrane transport proteins synthesis were significantly up-regulated. To sum up, the findings of this research will contribute to providing support for the application of L. brevis JLD715 in selenium-enriched functional foods. © 2021 Society of Chemical Industry.

Transcriptome analysis of azacitidine (5-AzaC)-treatment affecting the development of early somatic embryogenesis in longan

ABSTRACT DNA methylation plays key roles in regulating genes which control somatic embryogenesis (SE) in plant. 5-Azacytidine (5-AzaC) was considered as a common inhibitor of DNA methylation. Longan (Dimocarpus longan Lour.) SE has been established and used as model system for studying mechanism of SE in woody plants. However, effects of 5-AzaC on longan EC and the underlying mechanism at molecular level are rarely understood. In this study, RNA-seq was performed for transcriptome analysis of longan EC treated with or without 5-AzaC were analysed. A total of 29,232 expressed genes were obtained by aligned to longan reference genome and 1,617 differentially expressed genes (DEGs) were got in the pairwise comparisons of CK_vs_T. Integrating the KEGG enrichment pathway analysis showed that ‘butanoate metabolism’, ‘C5-branched dibasic acid metabolism’, ‘sulphur metabolism’, ‘selenocompound metabolism’, ‘pantothenate and CoA biosynthesis’, ‘phagosome’, and ‘plant hormone signal transduction’ were significantly enriched pathways, while the analysis of DEGs revealed that 5-AzaC-treatment could alter genes expression which related to the SE to adjust the complicated metabolic pathways. Therefore, these results indicated that 5-AzaC-treatment positively impact the development of early SE from longan and a decrease in the global DNA methylation level during early SE in longan.

Effects of Selenium- and Zinc-Enriched Lactobacillus plantarum SeZi on Antioxidant Capacities and Gut Microbiome in an ICR Mouse Model.

Selenium and zinc are essential trace minerals for humans with various biological functions. In this study, selenium- and zinc-tolerant lactic acid bacteria (LAB) isolates were screened out from human fecal samples. Amongst three hundred LAB isolates, the Lactobacillus plantarum SeZi strain displayed the tolerance against selenium and zinc with the greatest biomass production and bioaccumulation of selenium and zinc. To further assess the characteristics of this strain, the lyophilized L. plantarum SeZi were prepared and administered to Institute of Cancer Research (ICR) mice. The mice were divided into four groups, provided with normal chow (Con), or normal chow supplemented with Na2SeO3 and ZnSO4∙7H2O (SZ), L. plantarum SeZi (Lp), or selenium- and zinc-enriched L. plantarum SeZi (SZ + Lp), respectively. After 4 weeks of oral administration, the concentrations of selenium and zinc in blood were significantly increased in the SZ + Lp group when compared to the control or SZ group (p < 0.05). The increased selenium level led to an enhanced glutathione peroxidase activity and decreased blood malondialdehyde level in the SZ + Lp group (p < 0.05). Meanwhile, the results of bacterial community and microbial metabolic pathway analysis via 16S rRNA gene amplicon sequencing showed that L. plantarum SeZi significantly promoted the utilization of selenocysteine, seleno-cystathionine and seleno-methionine in the selenocompounds metabolism. Here, the in vivo antioxidant capacities of the selenium- and zinc-enriched lactobacillus strain showed us the utilization of a unique probiotic as a Se/Zn supplement with high availability, low toxicity, and additional probiotic advantages.

Mucosa-associated gut microbiome in Japanese patients with functional constipation.

The number of patients with chronic constipation is increasing in Japan. We investigated the gut mucosa-associated microbiome in Japanese patients with functional constipation. Diagnosis was made according to the Rome IV criteria. Mucosal samples were obtained by gentle brushing of mucosa surfaces. The gut microbiome was analyzed using 16S rRNA gene sequencing. There were no significant differences in bacteria α-diversity such as richness and evenness. The PCoA indicated significant structural differences between the constipation group and healthy controls (p=0.017 for unweighted and p=0.027 for weighted). The abundance of the phylum Bacteroidetes was significantly higher in the constipation group. The abundance of the genera Streptococcus, Fusobacterium, Comamonas, and Alistipes was significantly higher in the constipation group. The abundance of the genera Acinetobacter, Oscillospilla, Mucispirillum, Propinibacterium, and Anaerotruncus was significantly lower in the constipation group. In the constipation group, the proportion of genes responsible for sulfur metabolism, selenocompound metabolism, sulfur relay system was significantly higher and the proportion of d-arginine and d-ornithine metabolism and flavonoid biosynthesis was significantly lower. In conclusion, we identified differences of the mucosa-associated microbiome between Japanese patients with functional constipation and healthy controls. The mucosa-associated microbiome of functional constipation was characterized by higher levels of Bacteroidetes (Alistipes).

The integrative analysis of DNA methylation and mRNA expression profiles confirmed the role of selenocompound metabolism pathway in Kashin-Beck disease

ABSTRACT Kashin-Beck disease (KBD) is an endemic chronic osteochondropathy. The etiology of KBD remains unknown. In this study, we conducted an integrative analysis of genome-wide DNA methylation and mRNA expression profiles between KBD and normal controls to identify novel candidate genes and pathways for KBD. Articular cartilage samples from 17 grade III KBD patients and 17 healthy controls were used in this study. DNA methylation profiling of knee cartilage and mRNA expression profile data were obtained from our previous studies. InCroMAP was performed to integrative analysis of genome-wide DNA methylation profiles and mRNA expression profiles. Gene ontology (GO) enrichment analysis was conducted by online DAVID 6.7. The quantitative real-time polymerase chain reaction (qPCR), Western blot, immunohistochemistry (IHC), and lentiviral vector transfection were used to validate one of the identified pathways. We identified 298 common genes (such as COL4A1, HOXA13, TNFAIP6 and TGFBI), 36 GO terms (including collagen function, skeletal system development, growth factor), and 32 KEGG pathways associated with KBD (including Selenocompound metabolism pathway, PI3K-Akt signaling pathway, and TGF-beta signaling pathway). Our results suggest the dysfunction of many genes and pathways implicated in the pathogenesis of KBD, most importantly, both the integrative analysis and in vitro study in KBD cartilage highlighted the importance of selenocompound metabolism pathway in the pathogenesis of KBD for the first time.

Kolaviron modulates dysregulated metabolism in oxidative pancreatic injury and inhibits intestinal glucose absorption with concomitant stimulation of muscle glucose uptake

This present study investigated the antioxidative and antidiabetic properties of kolaviron by analysing its inhibitory effect on key metabolic activities linked to T2D, in vitro and ex vivo. Kolaviron significantly inhibited α-glucosidase and α-amylase activities, and intestinal glucose absorption dose-dependently, while promoting muscle glucose uptake. Induction of oxidative pancreatic injury significantly depleted glutathione level, superoxide dismutase, catalase, and ATPase activities, while elevating malondialdehyde and nitric oxide levels, acetylcholinesterase and chymotrypsin activities. These levels and activities were significantly reversed in tissues treated with kolaviron. Kolaviron depleted oxidative-induced metabolites, with concomitant restoration of oxidative-depleted metabolites. It also inactivated oxidative-induced ascorbate and aldarate metabolism, pentose and glucuronate interconversions, fructose and mannose metabolism, amino sugar and nucleotide sugar metabolism, and arginine and proline metabolism, while reactivating selenocompound metabolism. These results depict the antidiabetic properties of kolaviron as indicated by its ability to attenuate oxidative-induced enzyme activities and dysregulated metabolisms, and modulated the enzyme activities linked to hyperglycaemia.

Genome-wide CRISPR/Cas9 library screening to identify PAK1 as a critical driver for lenvatinib resistance in HCC.

e16687 Background: Lenvatinib demonstrated noninferiority to sorafenib in overall survival and a statistically significant increase in progression-free survival in patients with unresectable hepatocellular carcinoma (HCC). However, the development of drug resistance of lenvatinib is becoming common. Methods: Here, we performed the first genome-wide CRISPR/Cas9-based screening on lenvatinib-treated HCC cells to identify essential genes for non-mutational mechanisms related to acquired lenvatinib resistance and/or sensitivity in HCC cells. A genome-scale CRISPR knockout (mGeCKOa) library containing 123,411 single guide RNAs (sgRNAs) was used to identify loss-of-function mutations conferring lenvatinib resistance upon HCC cells. CCK8, wound-healing, transwell and flow cytometry assays were used. Subcutaneous xenotransplantation models were used to investigate the role in vivo. Results: By using genome-wide CRISPR/Cas9 library screening, we identify PAK1, a family member of serine/threonine p21-activating kinases, as a critical driver for lenvatinib resistance and the most-effective genetic suppressor of lenvatinib activity. Analysis of the survival of HCC after hepatic resection revealed that low PAK1 expression shortened overall and tumor-free survival (P &lt; 0.001). Further, loss of PAK1 from HCC cell increased the cytotoxicity of lenvatinib in vivo. Lenvatinib treatment activates series of metabolism, including “Glycolysis/ Gluconeogenesis、Cysteine and methionine metabolism、Selenocompound metabolism、Sphingolipid metabolism、Pyruvate metabolism、Metabolic pathways”. With RNAi knockdown and CRISPR/Cas9 knockout models, we show that loss of PAK1 activates the Glycolysis/ Gluconeogenesis and increase the production related metabolites. Concomitantly, activation of PAK1 reduces ROS level and HCC apoptosis upon Lenvatinib treatment. Lenvatinib treatment activates series of metabolism, including “Glycolysis/ Gluconeogenesis”. With RNAi knockdown and CRISPR/Cas9 knockout models, we show that loss of PAK1 activates the Glycolysis/ Gluconeogenesis and increase the production related metabolites. Concomitantly, activation of PAK1 reduces ROS level and HCC apoptosis upon Lenvatinib treatment. Conclusions: In summary, our results demonstrate that targeting PAK1 is an effective approach to overcome lenvatinib drug resistance in HCC.