Force-enhanced sensitive and specific detection of DNA-intercalative agents directly from microorganisms at single-molecule level.

Microorganisms can produce a vast array of bioactive secondary metabolites, including DNA-intercalating agents like actinomycin D, doxorubicin, which hold great potential for cancer chemotherapy. However, discovering novel DNA-intercalating compounds remains challenging due to the limited sensitivity and specificity of conventional activity assays, which require large-scale fermentation and purification. Here, we introduced the single-molecule stretching assay (SMSA) directly to microbial cultures or extracts for discovering DNA-intercalating agents, even in trace amounts of microbial cultures (5 µl). We showed that the unique changes of dsDNA in contour length and overstretching transition enable the specific detection of intercalators from complex samples without the need for extensive purification. Applying force to dsDNA also enhanced the sensitivity by increasing both the binding affinity Ka and the quantity of ligands intercalation, thus allowing the detection of weak intercalators, which are often overlooked using traditional methods. We demonstrated the effectiveness of SMSA, identified two DNA intercalator-producing strains: Streptomyces tanashiensis and Talaromyces funiculosus, and isolated three DNA intercalators: medermycin, kalafungin and ligustrone B. Interestingly, both medermycin and kalafungin, classified as weak DNA intercalators (Ka ∼103 M-1), exhibited potent anti-cancer activity against HCT-116 cancer cells, with IC50 values of 52 ± 6 and 70 ± 7 nM, respectively.

High serum uric acid may associate with the increased risk of colorectal cancer in females: A prospective cohort study.

Elevated serum uric acid (SUA) levels have been previously reported to play a role in multiple types of cancers. However, epidemiological studies evaluating SUA levels and colorectal cancer risk remain sparse. This cohort study included 444 462 participants between the ages of 40 and 69 years from the UK Biobank, followed up from 2006 to 2010. Multivariable adjusted Cox regression models were used to estimate hazard ratios (HRs). During a mean follow-up of 6.6 years, 2033 and 855 cases of colon and rectal cancers, respectively, were diagnosed. The multivariable-adjusted HRs for risks of colon cancer in the lowest uric acid categories (≤3.5 mg/dL) compared with the reference groups were 1.31 (95% confidence interval [CI] = 0.75-2.29) in males and 1.26 (95% CI = 1.03-1.55) in females. The HRs in the highest uric acid groups (>8.4 mg/dL) were 1.16 (95% CI = 0.83-1.63) in males and 2.00 (95% CI = 1.02-3.92) in females. The corresponding HRs of rectal cancer in the lowest uric acid groups compared with the reference group were 2.21 (95% CI = 1.15-4.23) in males and 0.98 (95% CI = 0.66-1.45) in females. The HRs in the highest uric acid groups were 1.35 (95% CI = 0.82-2.23) in males and 3.81 (95% CI = 1.38-10.56) in females. In conclusion, SUA showed a U-shaped association with colon cancer risk in both male and female populations. The same pattern was observed in male patients with rectal cancer. However, SUA levels were positively associated with occurrence of rectal cancer in female subjects.

A Review of Research on the Mechanism of Tumor Regulation by N-Acetyltransferase 10.

N-acetyltransferase 10 (NAT10) is an important acetyltransferase that regulates telomerase activity and participates in DNA damage reactions, ribosomal RNA (rRNA) transcriptional activation, cell division, microtubule acetylation, and other important cellular processes. Abnormalities in the expression or distribution of NAT10 result in diseases such as Hutchinson-Gilford progeria syndrome (HGPS) and various tumors, with serious consequences. Remodelin, an inhibitor of NAT10, delays HGPS progression; many studies have been conducted on its role in tumor therapy. A major breakthrough in the study of NAT10 was the discovery of mRNA N4-acetylcytidine (ac4C) modification, which can increase mRNA stability and translation efficiency significantly. In addition, NAT10 modifies the mRNA of ac4C, which is associated with tumor development. Here, we present a review of pertinent studies focusing on NAT10, particularly its role in cancer, to provide researchers with a concise and informative summary of the current state of knowledge about this topic. The conclusions drawn from this review could provide a new direction for tumor treatment.

Natural defence mechanisms of electrochemically active biofilms: From the perspective of microbial adaptation, survival strategies and antibiotic resistance.

Electrochemically active biofilms (EABs) play an ever-growingly critical role in the biological treatment of wastewater due to its low carbon footprint and sustainability. However, how the multispecies biofilms adapt, survive and become tolerant under acute and chronic toxicity such as antibiotic stress still remains well un-recognized. Here, the stress responses of EABs to tetracycline concentrations (CTC) and different operation schemes were comprehensively investigated. Results show that EABs can quickly adapt (start-up time is barely affected) to low CTC (≤ 5 µM) exposure while the adaptation time of EABs increases and the bioelectrocatalytic activity decreases at CTC ≥ 10 µM. EABs exhibit a good resilience and high anti-shocking capacity under chronic and acute TC stress, respectively. But chronic effects negatively affect the metabolic activity and extracellular electron transfer, and simultaneously change the spatial morphology and microbial community structure of EABs. Particularly, the typical exoelectrogens Geobacter anodireducens can be selectively enriched under chronic TC stress with relative abundance increasing from 45.11% to 85.96%, showing stronger TC tolerance than methanogens. This may be attributed to the effective survival strategies of EABs in response to TC stress, including antibiotic efflux regulated by tet(C) at the molecular level and the secretion of more extracellular proteins in the macro scale, as the C=O bond in amide I of aromatic amino acids plays a critical role in alleviating the damage of TC to cells. Overall, this study highlights the versatile defences of EABs in terms of microbial adaptation, survival strategies, and antibiotic resistance, and deepens the understanding of microbial communities' evolution of EABs in response to acute and chronic TC stress.

Surface energy-induced anti-wetting and anti-fouling enhancement of Janus membrane for membrane distillation.

Membrane distillation (MD) presents a promising alternative to conventional desalination systems, particularly for the treatment of hypersaline wastewater. However, the large-scale application of MD is hindered by challenges such as membrane wetting, membrane fouling, and low permeate flux. Herein, we proposed an air/liquid interface deposition method to fabricate a Janus membrane, termed the PVDF-PDA/PEI-Si membrane. The membrane featured a nanosieving, superhydrophilic polydopamine/polyethylenimine (PDA/PEI) layer decorated with silica nanoparticles, coupled with a microporous, hydrophobic polyvinylidene fluoride (PVDF) layer. The introduction of a dense PDA/PEI-Si layer featuring high surface energy significantly enhanced the wetting and fouling resistance of the membrane, with a minor effect on the permeate flux. The performance enhancement was particularly evident when hypersaline water containing sodium dodecyl sulfate (SDS) and oily contaminants was used as the feed. The interactions between the membrane and contaminants were calculated using the XDLVO theory and molecular dynamics simulations to elucidate the mechanisms underlying the enhanced anti-wetting and anti-fouling properties, respectively. According to the XDLVO theory, a large energy barrier must be overcome for the SDS to attach onto the PDA/PEI-Si surface. Meanwhile, molecular dynamics simulations confirmed the weak interaction energy between the oily foulants and the PVDF-PDA/PEI-Si membrane due to its high surface energy. This study presents a promising approach for the fabrication of high-performance MD membranes and provides new insights into the mechanisms underlying the enhanced anti-wetting and anti-fouling properties.

Sludge bound-EPS solubilization enhance CH4 bioconversion and membrane fouling mitigation in electrochemical anaerobic membrane bioreactor: Insights from continuous operation and interpretable machine learning algorithms.

Bound extracellular polymeric substances (EPS) are complex, high-molecular-weight polymer mixtures that play a critical role in pore clogging, foulants adhesion, and fouling layer formation during membrane filtration, owing to their adhesive properties and gelation tendencies. In this study, a novel electrochemical anaerobic membrane bioreactor (EC-AnMBR) was constructed to investigate the effect of sludge bound-EPS solubilization on methane bioconversion and membrane fouling mitigation. During the 150-days' operation, the EC-AnMBR demonstrated remarkable performance, characterized by an exceptionally low fouling rate (transmembrane pressure (TMP) < 4.0 kPa) and high-quality effluent (COD removal > 98.2 %, protein removal > 97.7 %, and polysaccharide removal > 98.5 %). The highest methane productivity was up to 38.0 ± 3.1 mL/Lreactor/d at the applied voltage of 0.8 V with bound-EPS solubilization, 107.6 % higher than that of the control stage (18.3 ± 2.4 mL/Lreactor/d). Morphological and multiplex fluorescence labeling analyses revealed higher fluorescence intensities of proteins, polysaccharides, total cells and lipids on the surface of the fouling layer. In contrast, the interior exhibited increased compression density and reduced activity, likely attributable to compression effect. Under the synergistic influence of the electric field and bound-EPS solubilization, biomass characteristics exhibited a reduced propensity for membrane fouling. Furthermore, the bio-electrochemical regulation enhanced the electroactivity of microbial aggregates and enriched functional microorganisms, thereby promoting biofilm growth and direct interspecies electron transfer. Additionally, the potential hydrogenotrophic and methylotrophic methanogenesis pathways were enhanced at the cathode and anode surfaces, thereby increasing CH4 productivity. The random forest-based machine learning model analyzed the nonlinear contributions of EPS characteristics on methane productivity and TMP values, achieving R² values of 0.879 and 0.848, respectively. Shapley additive explanations (SHAP) analysis indicated that S-EPSPS and S-EPSPN were the most critical factors affecting CH4 productivity and membrane fouling, respectively. Partial dependence plot analysis further verified the marginal and interaction effects of different EPS layers on these outcomes. By combining continuous operation with interpretable machine learning algorithms, this study unveils the intricate impacts of EPS characteristics on methane productivity and membrane fouling behaviors, and provides new insights into sludge bound-EPS solubilization in EC-AnMBR.

Enhanced dewaterability and triclosan removal of waste activated sludge with iron-rich mineral-activated peroxymonosulfate.

High water and pharmaceutical and care products (PPCPs) bounded in sludge flocs limit its utilization and disposal. The advanced oxidation process of perxymonosulfate (PMS) catalyzed by iron salts has been widely used in sludge conditioning. In this study, two iron-rich minerals pyrite and siderite were proposed to enhance sludge dewatering performance and remove the target contaminant of triclosan (TCS). The permanent release of Fe2+ in the activation of PMS made siderite more effective in enhancing sludge dewater with capillary suction time (CST) diminishing by 60.5 %, specific resistance to filtration (SRF) decreasing by 79.2 %, and bound water content (BWC) dropping from 37.1 % to 2.6 % at siderite/PMS dosages of 0.36/0.20 mmol/g-TSS after 20 min of pretreatment. Pyrite/PMS performed slightly inferior under the same conditions and the corresponding CST and SRF decreased by 51.5 % and 71.8 % while the BWC only declined to 17.8 %. Rheological characterization was employed to elucidate the changes in sludge dewatering performance, with siderite/PMS treated sludge showing a 48.3 % reduction in thixotropy, higher than 28.4 % of pyrite/PMS. Oscillation and creep tests further demonstrated the significantly weakened viscoelastic behavior of the sludge by siderite/PMS pretreatment. For TCS mineralization removal, siderite/PMS achieved a high removal efficiency of 43.9 %, in comparison with 39.9 % for pyrite/PMS. The reduction in the sludge solids phase contributed the most to the TCS removal. Free radical quenching assays and EPR spectroscopy showed that both siderite/PMS and pyrite/PMS produced SO4-·  and ·OH, with the latter acting as the major radicals. Besides, the dosage of free radicals generated from siderite/PMS exhibited a lower time-dependence, which also allowed it to outperform in destroying EPS matrix, neutralizing the negative Zeta potential of sludge flocs, and mineralizing macromolecular organic matter.

Helicobacter pylori infection alters gastric microbiota structure and biological functions in patients with gastric ulcer or duodenal ulcer.

BACKGROUND: Helicobacter pylori (H. pylori) infection is closely associated with gastrointestinal diseases. Our preliminary studies have indicated that H. pylori infection had a significant impact on the mucosal microbiome structure in patients with gastric ulcer (GU) or duodenal ulcer (DU). AIM: To investigate the contributions of H. pylori infection and the mucosal microbiome to the pathogenesis and progression of ulcerative diseases. METHODS: Patients with H. pylori infection and either GU or DU, and healthy individuals without H. pylori infection were included. Gastric or duodenal mucosal samples was obtained and subjected to metagenomic sequencing. The compositions of the microbial communities and their metabolic functions in the mucosal tissues were analyzed. RESULTS: Compared with that in the healthy individuals, the gastric mucosal microbiota in the H. pylori-positive patients with GU was dominated by H. pylori, with significantly reduced biodiversity. The intergroup differential functions, which were enriched in the H. pylori-positive GU patients, were all derived from H. pylori, particularly those concerning transfer RNA queuosine-modification and the synthesis of demethylmenaquinones or menaquinones. A significant enrichment of the uibE gene was detected in the synthesis pathway. There was no significant difference in microbial diversity between the H. pylori-positive DU patients and healthy controls. CONCLUSION: H. pylori infection significantly alters the gastric microbiota structure, diversity, and biological functions, which may be important contributing factors for GU.

Biogas upgrading and membrane anti-fouling mechanisms in electrochemical anaerobic membrane bioreactor (EC-AnMBR): Focusing on spatio-temporal distribution of metabolic functionality of microorganisms.

Electrochemical anaerobic membrane bioreactor (EC-AnMBR) by integrating a composite anodic membrane (CAM), represents an effective method for promoting methanogenic performance and mitigating membrane fouling. However, the development and formation of electroactive biofilm on CAM, and the spatio-temporal distribution of key functional microorganisms, especially the degradation mechanism of organic pollutants in metabolic pathways were not well documented. In this work, two AnMBR systems (EC-AnMBR and traditional AnMBR) were constructed and operated to identify the role of CAM in metabolic pathway on biogas upgrading and mitigation of membrane fouling. The methane yield of EC-AnMBR at HRT of 20 days was 217.1 ± 25.6 mL-CH4/g COD, about 32.1 % higher compared to the traditional AnMBR. The 16S rRNA analysis revealed that the EC-AnMBR significantly promoted the growth of hydrolysis bacteria (Lactobacillus and SJA-15) and methanogenic archaea (Methanosaeta and Methanobacterium). Metagenomic analysis revealed that the EC-AnMBR promotes the upregulation of functional genes involved in carbohydrate metabolism (gap and kor) and methane metabolism (mtr, mcr, and hdr), improving the degradation of soluble microbial products (SMPs)/extracellular polymeric substances (EPS) on the CAM and enhancing the methanogens activity on the cathode. Moreover, CAM biofilm exhibits heterogeneity in the degradation of organic pollutants along its vertical depth. The bacteria with high hydrolyzing ability accumulated in the upper part, driving the feedstock degradation for higher starch, sucrose and galactose metabolism. A three-dimensional mesh-like cake structure with larger pores was formed as a biofilter in the middle and lower part of CAM, where the electroactive Geobacter sulfurreducens had high capabilities to directly store and transfer electrons for the degradation of organic pollutants. This outcome will further contribute to the comprehension of the metabolic mechanisms of CAM module on membrane fouling control and organic solid waste treatment and disposal.

Controlled arrays of self-assembled peptide nanostructures in solution and at interface.

Controlling the formation of large and homogeneous arrays of bionanostructures through the self-assembly approach is still a great challenge. Here, we report the spontaneous formation of highly ordered arrays based on aligned peptide nanostructures in a solution as well as at an interface by self-assembly. By controlling the time and temperature of self-assembly in the solution, parallel fibrous alignments and more sophisticated two-dimensional "knitted" fibrous arrays could be formed from aligned rod-like fibers. During the formation of such arrays, the "disorder-to-order" transitions are controlled by the temperature-responsible motile short hydrophobic tails of the gemini-like amphiphilic peptides (GAPs) with asymmetric molecular conformation. In addition, the resulting long-range-ordered "knitted" fibrous arrays are able to direct mineralization of calcium phosphate to form organic-inorganic composite materials. In this study, the self-assembly behavior of these peptide building blocks at an interface was also studied. Highly ordered spatial arrays with vertically or horizontally aligned nanostructures such as nanofibers, microfibers, and microtubes could be formed through interfacial assembly. The regular structures and their alignments on the interface are controlled by the alkyl chain length of building blocks and the hydrophilicity/hydrophobicity property of the interface.

Long-term performance, microbial evolution and spatial microstructural characteristics of anammox granules in an upflow blanket filter (UBF) treating high-strength nitrogen wastewater.

Granule formation, microstructure and microbial spatial distribution are crucial to granule stability and nitrogen removal. Here, an upflow blanket filter (UBF) reactor with porous fixed cylinder carriers was fabricated and operated for 234 days to investigate overall performance and the formation mechanism of anammox granules. Results showed that the UBF performed the highest nitrogen removal efficiency of 93.19 ± 3.39% under nitrogen loading rate of 3.6 kg-N/m3/d and HRT of 2 h. The tryptophan-like proteins as the key component in EPS were vital for granules formation. Further 16 s rRNA analysis indicated that SBR1031 with a relative abundance of 40.5% played an important role in cell aggregation. Thus, anammox granules were developed successfully with a two-layered spatial structure where outer-layer was ammonia oxidizing bacteria and inner-core was anaerobic ammonia oxidizing bacteria. Together, introduction of porous fixed cylinder carriers is a valid method to avoid biomass loss and floatation.

Self-assembled electrochemically active biofilms doped with carbon nanotubes: Electron exchange efficiency and cytotoxicity evaluation.

Thick electrochemically active biofilms (EABs) will lead to insufficient extracellular electron transfer (EET) rate because of the limitation of both substrate diffusion and electron exchange. Herein, carbon nanotubes (CNTs)-doped EABs are developed through self-assembly. The highly conductive biofilms (internal resistance of ∼211 Ω) are efficiently enriched at CNTs dosage of 1 g L-1, with the stable power output of 0.568 W m-2 over three months. The embedded CNTs can act as electron tunnel to accelerate the EET rate in thick biofilm. Self-charging/discharging experiments and Nernst-Monod model stimulation demonstrate a higher net charge storage capacity (0.15 C m-2) and more negative half-saturation potential (-0.401 V) for the hybrid biofilms than that of the control (0.09 C m-2, and -0.378 V). Enzyme activity tests and the observation of confocal laser scanning microscopy by live/dead staining show a nearly negligible cytotoxicity of CNTs, and non-targeted metabonomics analysis reveals fourteen differential metabolites that do not play key roles in microbial central metabolic pathways according to KEGG compound database. The abundance of typical exoelectrogens Geobacter sp. is 2-fold of the control, resulting in a better bioelectrocatalytic activity. These finding provide a possible approach to prolong electron exchange and power output by developing a hybrid EABs doped with conductive material.

Interaction of Bmal1 and eIF2α/ATF4 pathway was involved in Shuxie compound alleviation of circadian rhythm disturbance-induced hepatic endoplasmic reticulum stress.

ETHNOPHARMACOLOGICAL RELEVANCE: Shuxie Compound (SX) combines the composition and efficacy of Suanzaoren decoction and Huanglian Wendan decoction. It can soothe the liver, regulate the qi, nourish the blood and calm the mind. It is used in the clinical treatment of sleep disorder with liver stagnation. Modern studies have proved that circadian rhythm disorder (CRD) can cause sleep deprivation and liver damage, which can be effectively ameliorated by traditional Chinese medicine to soothe the liver stagnation. However, the mechanism of SX is unclear. AIM OF THE STUDY: This study was designed to demonstrate the impact of SX on CRD in vivo, and confirm the molecular mechanisms of SX in vitro. MATERIALS AND METHODS: The quality of SX and drug-containing serum was controlled by UPLC-Q-TOF/MS, which were used in vivo and in vitro experiments, respectively. In vivo, a light deprivation mouse model was used. In vitro, a stable knockdown Bmal1 cell line was used to explore SX mechanism. RESULTS: Low-dose SX (SXL) could restore (1) circadian activity pattern, (2) 24-h basal metabolic pattern, (3) liver injury, and (4) Endoplasmic reticulum (ER) stress in CRD mice. CRD decreased the liver Bmal1 protein at ZT15, which was reversed by SXL treatment. Besides, SXL decreased the mRNA expression of Grp78/ATF4/Chop and the protein expression of ATF4/Chop at ZT11. In vitro experiments, SX reduced the protein expression of thapsigargin (tg)-induced p-eIF2α/ATF4 pathway and increase the viability of AML12 cells by increasing the expression of Bmal1 protein. CONCLUSIONS: SXL relieved CRD-induced ER stress and improve cell viability by up-regulating the expression of Bmal1 protein in the liver and then inhibiting the protein expression of p-eIF2α/ATF4.

Shu-Xie decoction alleviates oxidative stress and colon injury in acute sleep-deprived mice by suppressing p62/KEAP1/NRF2/HO1/NQO1 signaling.

Introduction: Sleep disorders are common clinical psychosomatic disorders that can co-exist with a variety of conditions. In humans and animal models, sleep deprivation (SD) is closely related with gastrointestinal diseases. Shu-Xie Decoction (SX) is a traditional Chinese medicine (TCM) with anti-nociceptive, anti-inflammatory, and antidepressant properties. SX is effective in the clinic for treating patients with abnormal sleep and/or gastrointestinal disorders, but the underlying mechanisms are not known. This study investigated the mechanisms by which SX alleviates SD-induced colon injury in vivo. Methods: C57BL/6 mice were placed on an automated sleep deprivation system for 72 h to generate an acute sleep deprivation (ASD) model, and low-dose SX (SXL), high-dose SX (SXH), or S-zopiclone (S-z) as a positive control using the oral gavage were given during the whole ASD-induced period for one time each day. The colon length was measured and the colon morphology was visualized using hematoxylin and eosin (H&E) staining. ROS and the redox biomarkers include reduced glutathione (GSH), malondialdehyde (MDA), and superoxide dismutase (SOD) were detected. Quantitative real-time PCR (qRT-PCR), molecular docking, immunofluorescence and western blotting assays were performed to detect the antioxidant signaling pathways. Results: ASD significantly increased FBG levels, decreased colon length, moderately increased the infiltration of inflammatory cells in the colon mucosa, altered the colon mucosal structure, increased the levels of ROS, GSH, MDA, and SOD activity compared with the controls. These adverse effects were significantly alleviated by SX treatment. ASD induced nuclear translocation of NRF2 in the colon mucosal cells and increased the expression levels of p62, NQO1, and HO1 transcripts and proteins, but these effects were reversed by SX treatment. Conclusion: SX decoction ameliorated ASD-induced oxidative stress and colon injury by suppressing the p62/KEAP1/NRF2/HO1/NQO1 signaling pathway. In conclusion, combined clinical experience, SX may be a promising drug for sleep disorder combined with colitis.

Insights into biodegradation behaviors of methanolic wastewater in up-flow anaerobic sludge bed (UASB) reactor coupled with in-situ bioelectrocatalysis.

Granular sludge disintegration and washing out pose a challenge to up-flow anaerobic sludge bed (UASB) reactor treating methanolic wastewater. Herein, in-situ bioelectrocatalysis (BE) was integrated into UASB (BE-UASB) reactor to alter microbial metabolic behaviors and enhance the re-granulation process. BE-UASB reactor exhibited the highest methane (CH4) production rate of 388.0 mL/Lreactor/d and chemical oxygen demand (COD) removal of 89.6 % at 0.8 V. Sludge re-granulation was strengthened with particle size over 300 µm of up to 22.4%. Bioelectrocatalysis stimulated extracellular polymeric substances (EPS) secretion and formation of granules with rigid [-EPS-cell-EPS-] matrix by enhancing the proliferation of key functional microorganisms (Acetobacterium, Methanobacterium, and Methanomethylovorans) and diversifying metabolic pathways. Particularly, a high Methanobacterium richness (10.8%) drove the electroreduction of CO2 into CH4 and reduced its emissions (52.8%). This study provides a novel bioelectrocatalytic strategy for controlling granular sludge disintegration, which will facilitate the practical application of UASB in methanolic wastewater treatment.

Corrigendum: Shu-Xie decoction alleviates oxidative stress and colon injury in acute sleep-deprived mice by suppressing p62/KEAP1/NRF2/HO1/NQO1 signaling.

[This corrects the article DOI: 10.3389/fphar.2023.1107507.].

Impact of sandwich-type composite anodic membrane on membrane fouling and methane recovery from sewage sludge and food waste via electrochemical anaerobic membrane bioreactor.

Membrane fouling presents a big challenge for the real-world implementation of anaerobic membrane bioreactors (AnMBRs) in digesting high-solid biowastes. In this study, an electrochemical anaerobic membrane bioreactor (EC-AnMBR) with a novel sandwich-type composite anodic membrane was designed and constructed for controlling membrane fouling whilst improving the energy recovery. The results showed that EC-AnMBR produced a higher methane yield of 358.5 ± 74.8 mL/d, rising by 12.8% compared to the AnMBR without applied voltage. Integration of composite anodic membrane induced a stable membrane flux and low transmembrane pressure through forming an anodic biofilm while total coliforms removal reached 97.9%. The microbial community analysis further provided compelling evidence that EC-AnMBR enriched the relative abundance of hydrolyzing (Chryseobacterium 2.6%) bacteria and methane-producing (Methanobacterium 32.8%) archaea. These findings offered new insights into anti-biofouling performance and provided significant implications for municipal organic waste treatment and energy recovery in the new EC-AnMBR.

Diversity of Fungal Communities on Diseased and Healthy Cinnamomum burmannii Fruits and Antibacterial Activity of Secondary Metabolites.

The composition and structure of fungal communities on healthy and diseased fruits of Cinnamomum burmannii (Nees and Nees) Blume were characterized, with evaluation of the antibacterial activity of secondary metabolites from culturable fungi following the first identification of secondary metabolites in the fungus Medicopsis romeroi (Esf-14; GenBank accession number OK242756). These results are significant for understanding the functional variation in bioactivity in fungal communities and developing a broader range of bioactive resources. High-throughput sequencing results indicated that the fungal community in diseased fruit differed from that in healthy fruit at the phylum, class, order, or genus level, with significant differences in the species and relative abundance of the dominant flora. A total of 49 (healthy fruit) and 122 (diseased fruit) artificially cultivable endophytic fungi were isolated, and 41 different strains (11 from healthy fruit and 30 from diseased fruit) were successfully identified by morphological and molecular biological analyses, which were classified into 8 groups and 23 genera by phylogenetic tree analysis, with Pleosporales, Glomerellales, and Hypocreales being the dominant groups at the order level and Colletotrichum being the dominant group at the genus level. The results of the antibacterial assay demonstrated that the secondary metabolites of all strains had different degrees of antibacterial activity, while the secondary metabolites of endophytic fungi from diseased fruit were generally stronger than those of fungi from healthy fruit, with the active secondary metabolites dominated by small and moderately polar compounds. Combined analysis of fungal communities, phylogenetic tree analysis, and bioactivity analysis of culturable strains revealed strong antibacterial activity of both upregulated and downregulated flora in diseased fruit. Five compounds, including two new (5,6-dimethoxy-[1',1:4,1″-terphenyl]-2-ol [compound 1] and 5-(methoxycarbonyl)-2-methylbenzo[d][1,3]dioxole-2-carboxylic acid [compound 2]) and three known compounds (3,7-dihydroxy-1,9-dimethyldibenzofuran [compound 3], methyl 3-hydroxybenzoate [compound 4], and uracil [compound 5]), were isolated and identified for the first time from the endophytic fungus Medicopsis romeroi. In general, the diversity of fungal communities on diseased fruit was lower than that on healthy fruits, while the antibacterial activity of artificially cultured endophytic fungi on diseased fruits was generally stronger than that on healthy fruits, suggesting excellent promise for the development of secondary metabolites from active strains on diseased fruit as antibacterial agents. IMPORTANCE Powdery fruit disease is a notorious disease of Cinnamomum burmannii that causes severe loss in fruit production. Studies on the function of endophytic fungal communities in healthy plant tissues are not new, while little is known about the functional changes of fungal communities in disease-causing plant tissues. Our results demonstrate that fungal communities in diseased fruits differ from those in healthy fruits at the level of phylum, class, order, or genus, with significant differences in the species and relative abundance of dominant groups. Endophytic fungi in diseased fruits appeared to produce secondary metabolites with stronger antibacterial properties, although the community diversity was not as varied as that in healthy fruits. In addition, secondary metabolites of the Medicopsis romeroi strain from diseased fruits were identified for the first time. These results have important implications for understanding the functional variation of bioactivity in fungal communities and for developing a broader resource of bioactivity.

Lanatoside C decelerates proliferation and induces apoptosis through inhibition of STAT3 and ROS-mediated mitochondrial membrane potential transformation in cholangiocarcinoma.

Introduction: The incidence of cholangiocarcinoma (CCA) has increased worldwide in recent years. Given the poor prognosis associated with the current management approach of CCA, new therapeutic agents are warranted to improve the prognosis of this patient population. Methods: In this study, we extracted five cardiac glycosides (CGs) from natural plants: digoxin, lanatoside A, lanatoside C, lanatoside B, and gitoxin. Follow-up experiments were performed to assess the effect of these five extracts on cholangiocarcinoma cells and compounds with the best efficacy were selected. Lanatoside C (Lan C) was selected as the most potent natural extract for subsequent experiments. We explored the potential mechanism underlying the anticancer activity of Lan C on cholangiocarcinoma cells by flow cytometry, western blot, immunofluorescence, transcriptomics sequencing, network pharmacology and in vivo experiments. Results: We found that Lan C time-dependently inhibited the growth and induced apoptosis of HuCCT-1 and TFK-1 cholangiocarcinoma cells. Besides Lan C increased the reactive oxygen species (ROS) content in cholangiocarcinoma cells, decreased the mitochondrial membrane potential (MMP) and resulted in apoptosis. Besides, Lan C downregulated the protein expression of STAT3, leading to decreased expression of Bcl-2 and Bcl-xl, increased expression of Bax, activation of caspase-3, and initiation of apoptosis. N-acetyl-L-cysteine (NAC) pretreatment reversed the effect of Lan C. In vivo, we found that Lan C inhibited the growth of cholangiocarcinoma xenografts without toxic effects on normal cells. Tumor immunohistochemistry showed that nude mice transplanted with human cholangiocarcinoma cells treated with Lan C exhibited decreased STAT3 expression and increased caspase-9 and caspase-3 expression in tumors, consistent with the in vitro results. Conclusion: In summary, our results substantiates that cardiac glycosides have strong anti-CCA effects. Interestingly the biological activity of Lan C provides a new anticancer candidate for the treatment of cholangiocarcinoma.

[Advances in the study of vasoactive-inotropic score in critically ill patients].

The degree of hemodynamic support by vasoactive drugs in critically ill patients is often considered one of the markers of disease severity. The sequential organ failure assessment (SOFA), European system for cardiac operative risk evaluation II (EuroScore II), and other scores only roughly quantify the drug support of cardiovascular system. When patients need large doses of vasoactive drugs, the mortality increases accordingly. The vasoactive-inotropic score (VIS) objectively quantifies the degree of cardiovascular support using a simple formula that standardizes the dose of different agents, and it is recommended as a simple, effective, and accurate prognostic indicator. In recent years, there are more and more clinical applications and related studies at home and abroad. This paper reviews the application and progress of VIS score in critically ill patients, providing help for doctors to judge the condition and prognosis of patients and guiding the decision-making of diagnosis and treatment.