Extracellular Protein Concentration Research Articles

Revealing detrimental effects of various DC electrical energy conditions on different multidrug resistant bacteria: a comprehensive study

The arbitrary discharge of contaminated wastes, especially that encompass multidrug resistant microbes (MDR), would broaden the circle of epidemic diseases such as COVID-19, which in turn deteriorate definitely the whole socioeconomics. Therefore, the employment of electrical stimulation techniques such as direct current (DC) with low energy considers being effective tool to impede spontaneous changes in microbial genetic makeup, which increases the prevalence of MDR phenomenon. Herein, the influence of different electric energies generated by DC electric field, volts and time on MDR-bacteria that are categorized among the highly ranked nosocomial pathogens, was scrutinized. Wherein, Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Enterococcus faecalis were examined as paradigms of Gram-negative and Gram-positive pathogens. The results declared the significant superior antagonizing potency of electric energy in a dose-dependent modality rather than the applied volts or exposure time. Notably, the exposure of bacterial cultures to140 J inhibited the bacterial count by > 78% and the range of 47–73% for Gram-negative and Gram-positive, respectively. While the suppression in their metabolic activity assessed by > 75% and 41–68%, respectively; reflecting the capability of electrical energy to induce viable but non-culturable (VBNC) state. Similarly, the results of total protein, extracellular protein content and lactate dehydrogenase activity emphasized the cell wall deterioration and losing of cell membrane integrity. Additionally, the elevating in ROS upon DC-exposure participated in DNA fragmentation and plasmid decomposability by the range of 33–60%. Further, SEM micrographs depicted drastic morphological deformations after electrical treatment. Strikingly, DC-treatment impaired antibiotic resistance of the examined strains against several antibiotics by > 64.2%. Generally, our comparative detailed study revealed deleterious potentiality of different DC-protocols in defeating microbial pollution, which could be invested as efficient disinfectant alternative in various sectors such as milk sterilization and wastewater purification.

Enhanced biodegradation of benzo[a]pyrene with Trametes versicolor stimulated by citric acid.

Polycyclic aromatic hydrocarbons (PAHs) are a class of persistent organic pollutants with carcinogenic, mutagenic and teratogenic effects. The white-rot fungi in the fungal group have significant degradation ability for high molecular weight organic pollutants. However, exogenous fungi are easily antagonized by indigenous microorganisms. Low molecular weight organic acids, a small molecular organic matter secreted by plants, can provide carbon sources for soil microorganisms. Combining organic acids with white rot fungi may improve the nutritional environment of fungi. In this study, immobilized Trametes versicolor was used to degrade benzo[a]pyrene in soil, and its effect on removing benzo[a]pyrene in soil mediated by different low molecular weight organic acids was investigated. The results showed that when the degradation was 35days, the removal effect of the experimental group with citric acid was the best, reaching 43.7%. The degradation effect of Trametes versicolor on benzo[a]pyrene was further investigated in the liquid medium when citric acid was added, and the effects of citric acid on the biomass, extracellular protein concentration and laccase activity of Trametes versicolor were investigated by controlling different concentrations of citric acid. In general, citric acid can act as a carbon source for Trametes versicolor and promote its extracellular protein secretion and laccase activity, thereby accelerating the mineralization of benzo[a]pyrene by Trametes versicolor. Therefore, citric acid can be used as a biostimulant in the remediation of PAHs contaminated soil with Trametes versicolor.

Sodium acetate increases the productivity of HEK293 cells expressing the ECD-Her1 protein in batch cultures: experimental results and metabolic flux analysis.

Human Embryonic Kidney cells (HEK293) are a popular host for recombinant protein expression and production in the biotechnological industry. This has driven within both, the scientific and the engineering communities, the search for strategies to increase their protein productivity. The present work is inserted into this search exploring the impact of adding sodium acetate (NaAc) into a batch culture of HEK293 cells. We monitored, as a function of time, the cell density, many external metabolites, and the supernatant concentration of the heterologous extra-cellular domain ECD-Her1 protein, a protein used to produce a candidate prostate cancer vaccine. We observed that by adding different concentrations of NaAc (0, 4, 6 and 8mM), the production of ECD-Her1 protein increases consistently with increasing concentration, whereas the carrying capacity of the medium decreases. To understand these results we exploited a combination of experimental and computational techniques. Metabolic Flux Analysis (MFA) was used to infer intracellular metabolic fluxes from the concentration of external metabolites. Moreover, we measured independently the extracellular acidification rate and oxygen consumption rate of the cells. Both approaches support the idea that the addition of NaAc to the culture has a significant impact on the metabolism of the HEK293 cells and that, if properly tuned, enhances the productivity of the heterologous ECD-Her1 protein.

Enhanced bioremediation of organically combined contaminated soil by white rot fungal agent: physiological characteristics and contaminants degradation

ABSTRACT Microbial remediation of organically combined contaminated sites is currently facing technical challenges. White rot fungi possess broad-spectrum degradation capabilities, but most of the studies are conducted on polluted water bodies, and few research focus on the degradation of combined organically contaminated soils. This study aimed to investigate the physiological changes in Trametes versicolor to enhance its simultaneous degradation ability towards benzo(a)pyrene (BaP) and TPH. The results demonstrated that Trametes versicolor, when subjected to liquid fermentation, achieved an 88.08% degradation of individual BaP within 7 days. However, under the combined contamination conditions of BaP and TPH, the BaP degradation rate decreased to 69.25%, while the TPH degradation rate was only 16.95%. Furthermore, the degradation rate of BaP exhibited a significant correlation with the extracellular protein concentration and laccase activities. Conversely, the TPH degradation rate exhibited a significant and positive correlation with the intracellular protein concentration. Solid-state fermentation utilizing fungal agents proved to be the most effective method for removing BaP and TPH, yielding degradation rates of 56.16% and 15.73% respectively within 60 days. Overall, Trametes versicolor demonstrated a commendable capability for degrading combined PAHs-TPH pollutants, thereby providing theoretical insights and technical support for the remediation of organically combined contaminated sites.

Extracellular Vesicle Protein Expression in Doped Bioactive Glasses: Further Insights Applying Anomaly Detection.

Proteomic analysis of extracellular vesicles presents several challenges due to the unique nature of these small membrane-bound structures. Alternative analyses could reveal outcomes hidden from standard statistics to explore and develop potential new biological hypotheses that may have been overlooked during the initial evaluation of the data. An analysis sequence focusing on deviating protein expressions from donors' primary cells was performed, leveraging machine-learning techniques to analyze small datasets, and it has been applied to evaluate extracellular vesicles' protein content gathered from mesenchymal stem cells cultured on bioactive glass discs doped or not with metal ions. The goal was to provide additional opportunities for detecting details between experimental conditions that are not entirely revealed with classic statistical inference, offering further insights regarding the experimental design and assisting the researchers in interpreting the outcomes. The methodology extracted a set of EV-related proteins whose differences between conditions could be partially explainable with statistics, suggesting the presence of other factors involved in the bioactive glasses' interactions with tissues. Outlier identification of extracellular vesicles' protein expression levels related to biomaterial preparation was instrumental in improving the interpretation of the experimental outcomes.

Metabolomics-based analysis of the interaction effects and stress response mechanisms in the co-culture system of fungi and microalgae under cadmium stress

Research on the co-culture of fungi and microalgae to harvest microalgae and enhance the adsorption capacity for heavy metals to reduce contamination is well demonstrated, but the study of the correlation mechanism among critical metabolites, system stability, and stress response in the system under cadmium stress based on metabolomics analyses has not been reported in detail. In this study, the co-culture system of Aspergillus allahabadii and Synechocystis sp. PCC6803 was taken as the object of investigation, and isotope labelling (13C–Na2CO3 and 15N-(NH4)2SO4) and nanoscale secondary ion mass spectrometry were used to confirm the existence of carbon and nitrogen nutrient exchange. The adsorption behavior of the system after Cd treatment was investigated by Fourier Transform Infrared Spectroscopy, X-ray Photoelectron Spectroscopy, and Scanning Electron Microscope/Transmission Electron Microscope. The content of extracellular polysaccharides in the system was increased from 40.8 mg/g to 80.12 mg/g, and the content of extracellular proteins was increased from 22.3 mg/g to 55.68 mg/g after 96 h of Cd treatment. Metabolomics results revealed 235 differential metabolites co-expressed after Cd treatment, with 95, 15, and 77 unique differential metabolites. In particular, the synthesis of extracellular polymeric substances was a factor affecting the stability of the symbiotic system, the up-regulation of the expression of the transporter system was one of the reasons for the improvement of the system's tolerance and resistance to heavy metals; and the nutrient metabolism between the fungus and the microalgae was the main defense mechanism to reduce the extent of the impairment of the system's carbon sequestration capacity by Cd. This provides a theoretical basis for the green and economically friendly application of the symbiotic system in microalgae harvesting, efficient carbon dioxide fixation and reduction of heavy metal pollution.

Antibacterial Activity and Mechanism of Self-Assembly Spermidine-Capped Carbon Dots against Staphylococcus aureus.

This paper investigated the antibacterial mechanism of spermidine-capped carbon dots (S-PCDs) against Staphylococcus aureus. The results showed that there were a large number of amino groups on the surface of S-PCDs and they had a high positive charge (+47.06 mV), which could be adsorbed on the negatively charged bacterial surface through electrostatic interaction and changed the permeability of the bacterial cell membrane. The extracellular protein and nucleic acid contents of S. aureus treated with S-PCDs were 5.4 and 1.2 times higher than those of the control group, respectively. The surface folds and defects of the bacterial cell membrane, and the leakage of cell contents were observed using SEM and TEM. The expression of metabolic oxidation regulatory genes dmpI, narJ and narK was upregulated and the intracellular ROS generation was induced, causing bacterial oxidative stress and eventually bacterial death. S-PCDs can effectively inhibit biofilm formation and had low cytotoxicity. The S-PCD treatment successfully inhibited microbial reproduction when pasteurized milk was stored at 25 °C and 4 °C. These results provide important insights into the antimicrobial mechanism of S-PCDs and lay the foundation for their application in the food field as a potentially novel bacteriostatic nanomaterial.

Regulation of σB-Dependent Biofilm Formation in Staphylococcus aureus through Strain-Specific Signaling Induced by Diosgenin.

Staphylococcus aureus is a commensal skin bacterium and a causative agent of infectious diseases. Biofilm formation in S. aureus is a mechanism that facilitates the emergence of resistant strains. This study proposes a mechanism for the regulation of biofilm formation in S. aureus through strain-specific physiological changes induced by the plant steroid diosgenin. A comparison of diosgenin-induced changes in the expression of regulatory genes associated with physiological changes revealed the intracellular regulatory mechanisms involved in biofilm formation. Diosgenin reduced biofilm formation in S. aureus ATCC 6538 and methicillin-resistant S. aureus (MRSA) CCARM 3090 by 39% and 61%, respectively. Conversely, it increased biofilm formation in S. aureus ATCC 29213 and MRSA CCARM 3820 by 186% and 582%, respectively. Cell surface hydrophobicity and extracellular protein and carbohydrate contents changed in a strain-specific manner in response to biofilm formation. An assessment of the changes in gene expression associated with biofilm formation revealed that diosgenin treatment decreased the expression of icaA and spa and increased the expression of RNAIII, agrA, sarA, and sigB in S. aureus ATCC 6538 and MRSA CCARM 3090; however, contrasting gene expression changes were noted in S. aureus ATCC 29213 and MRSA CCARM 3820. These results suggest that a regulatory mechanism of biofilm formation is that activated sigB expression sequentially increases the expression of sarA, agrA, and RNAIII. This increased RNAIII expression decreases the expression of spa, a surface-associated adhesion factor. An additional regulatory mechanism of biofilm formation is that activated sigB expression decreases the expression of an unknown regulator that increases the expression of icaA. This in turn decreases the expression of icaA, which decreases the synthesis of polysaccharide intercellular adhesins and ultimately inhibits biofilm formation. By assessing strain-specific contrasting regulatory signals induced by diosgenin in S. aureus without gene mutation, this study elucidated the signal transduction mechanisms that regulate biofilm formation based on physiological and gene expression changes.

Biofilm formation and development during the pyrite bioleaching of moderately thermophilic microorganisms

Bioleaching is a process that uses microorganisms to recover metals from intractable metal sulfide ores. Metal sulfide bioleaching is an interfacial process, and biofilm growth is crucial during the early phases. To study the interactions between microbes and minerals during the bioleaching of moderately thermophilic microorganisms, the formation and development of biofilms on pyrite coupons incubated in shake flasks for 40 days were evaluated. The biofilm biomass progressively increased as bioleaching proceeded. The most extensive biofilm was formed on day 30, when bacteria were attached to the mineral surface or submerged in extracellular polymeric substances. Confocal laser scanning microscopy indicated that the biofilm biomass of pyrite on day 40 was 6.55, 5.28, 3.17, and 1.70 times higher than that on days 5, 10, 20, and 30, respectively. Extracellular protein content exhibited an increasing trend throughout the process, whereas extracellular polysaccharides played a significant role during the early stages. Live/dead staining showed the highest mortality rate of 94% when cells were in contact with pyrite in the early stage, and the percentage of live bacteria increased in the later period. In the bioleaching environment, mature biofilms appeared to form relatively quickly after day 20. Real-time quantitative fluorescent quantitative analysis indicated that biofilm formation played an important role in defense against hostile external environments. In this study, biofilm formation and the associated RNA transcript numbers of moderately thermophilic microorganisms during pyrite bioleaching were investigated, contributing to the understanding of cell-mineral interactions.

Effects of flow velocity on biofilm composition and microbial molecular ecological network in reclaimed water distribution systems

The existence of biofilm on the reclaimed water pipeline seriously affects the safety of water distribution. And the flow regimes in the pipeline play a crucial role in the growth of biofilms. In this study, the biofilm composition, surface topography and bacterial community were detected under eight levels of flow velocity in the range of 0.10–1.40 m s−1. The results showed that the dry weight, the concentration of extracellular protein and extracellular polysaccharide in the biofilm reached a dynamic stable period after 640 h. The biofilm composition and surface topography of biofilm were significantly different under the different flow regimes (laminar flow belongs to [0.10, 0.19] m s−1, and turbulent flow belongs to [0.29, 1.40] m s−1). As the flow velocity range increases, the concentration of each component in the biofilm and the parameters of biofilm surface topography increased and then decreased. The flow velocity could be a strong environmental stimulus resulting in the succession of bacterial community in biofilm. As the flow velocity increased from 0.10 m s−1 to 1.40 m s−1, at the phylum level, the average relative abundance of Firmicutes mainly showed a trend of first increasing and then decreasing with the highest abundance value of 71.57% at 0.49 m s−1. The flow velocity increased from 0.10 m s−1 to 0.49 m s−1, a significant increase in microbial diversity could be detected. The increase in flow velocity promoted the proliferation of microorganisms, and the interaction between different microbial components was enhanced. At 0.49 m s−1, the function of the biofilm is complex, and the ability to resist environmental stress is the strongest. This study can effectively improve the cognition depth of biofilms under the influence of flow velocity in the reclaimed water distribution systems, and provide an important theoretical support for the safe distribution of reclaimed water.

Growth, tolerance, and enzyme activities of Trichoderma strains in culture media added with a pyrethroids-based insecticide

The application of pyrethroids and carbamates represents an environmental risk and may exert adverse effects on beneficial microorganisms such as Trichoderma, which contribute to the biocontrol of several fungal phytopathogens. This research evaluated the tolerance of several strains of Trichoderma to a selected culture medium contaminated with a commercial insecticide (H24®) composed of pyrethroids, permethrin and prallethrin, and carbamate propoxur, and determined the influence of this insecticide on the release of enzymes such as chitinases, peroxidases, and endoglucanases by a consortium of selected Trichoderma strains grown in liquid culture medium. Four out of 10 Trichoderma strains showed tolerance to 200ppm (∼48.3% of growth) of the commercial insecticide after 96h of exposure to a contaminated solid medium. After eight days of growth in liquid culture, the insecticide enhanced extracellular protein content and peroxidase activities in the Trichoderma consortium but decreased both chitinase and glucanase activities. These fungal responses should be considered when implementing strategies that combine alternative pesticides and fungal biocontrollers for managing fungal phytopathogens.

Effect of the zinc transporter ZupT on the virulence mechanisms of mesophilic Aeromonas salmonicida SRW‐OG1

AbstractAs a typical psychrophilic bacterial pathogen, Aeromonas salmonicida causes furunculosis in wild and farmed freshwater and marine fish, leading to substantial economic losses in the global aquaculture industry. Previous studies have shown that A. salmonicida is unable to grow above 25°C, hence limiting its infection to cold‐water fish. However, we isolated A. salmonicida SRW‐OG1, a mesophilic pathogenic strain from the warm‐water fish Epinephelus coioides. Through RNA‐seq analysis, we observe significant upregulation of the zupT gene at 28°C. ZupT is a member of zinc‐regulated transporters and iron‐regulated transporter‐like proteins (ZIP family) and is closely associated with transcriptional regulation of virulence in certain pathogens. Consequently, our study aimed to examine the role of zupT during A. salmonicida SRW‐OG1 infection at high temperatures. Our findings demonstrate that the zupT‐RNAi strain exhibits severe growth restriction under limited Zn2+ and Fe2+ conditions. Notably, this strain shows significantly reduced mortality rates and colonization abilities. Moreover, its motility, biofilm formation, adhesion, and hemolytic activities are significantly diminished. Confocal laser scanning microscopy reveals earlier and accelerated biofilm dissociation in the zupT‐RNAi strain. Analysis of extracellular products at 36 h indicates a considerable reduction in relative extracellular protein content in the zupT‐RNAi strain. Taken together, our results highlight the vital role of the zupT gene in zinc transport and the fitness of A. salmonicida SRW‐OG1 within the host.

Absence of nitrogen and phosphorus in activated sludge: Impacts on flocculation characteristics and the microbial community

A sequencing batch reactor operated in anoxic/aerobic mode was used to investigate the effects of simultaneous nitrogen (N) and phosphorus (P) absence in activated sludge. Through controlling the N and P content in the influent water, we investigated changes in sludge concentration, settling performance, morphological structure, surface properties, extracellular polymer content, and microbial community structure and function. When N and P were simultaneously absent, the relative hydrophobicity of the activated sludge and the sludge concentration decreased. The specific oxygen uptake rate also decreased, the activated sludge did not bulk, and the flocs were larger and more concentrated. The content of extracellular polysaccharides did not significantly change. An increase in the concentration of extracellular proteins, particularly loose-binding layer proteins, improved activated sludge aggregation. The structure and function of the activated sludge microbial community changed significantly in the absence of both N and P, with a significant decrease in the relative abundance of species related to N and P metabolism. Tax4Fun analysis showed that the absence of both N and P decreased metabolism and genetic information processing of the microbial community. These results elucidate the effects of simultaneous N and P absence on activated sludge from the perspective of microbial community structure and function. These results will be useful for treatment of N and P deficient wastewater by the activated sludge method.

Measuring (biological) materials mechanics with atomic force microscopy. 5. Traction force microscopy (cell traction forces).

Cells generate traction forces to probe the mechanical properties of the surroundings and maintain a basal equilibrium state of stress. Traction forces are also implicated in cell migration, adhesion and ECM remodeling, and alteration of these forces is often observed in pathologies such as cancer. Thus, analyzing the traction forces is important for studies of cell mechanics in cancer and metastasis. In this primer, the methodology for conducting two-dimensional traction force microscopy (2D-TFM) experiments is reported. As a practical example, we analyzed the traction forces generated by three human breast cancer cell lines of different metastatic potential: MCF10-A, MCF-7 and MDA-MB-231 cells, and studied the effects of actin cytoskeleton disruption on those traction forces. Contrary to what is often reported in literature, lower traction forces were observed in cells with higher metastatic potential (MDA-MB-231). Implications of substrate stiffness and concentration of extracellular matrix proteins in such findings are discussed in the text. RESEARCH HIGHLIGHTS: Traction force microscopy (TFM) is suitable for studying and quantifying cell-substrate and cell-cell forces. TFM is suitable for investigating the relationship between chemical to mechanical signal transduction and vice versa. TFM can be combined with classical indentation studies providing a compact picture of cell mechanics. TFM still needs new physico-chemical (sample preparation) and computational approaches for more accurate data evaluation.

Development potential of extracellular matrix hydrogels as hemostatic materials.

The entry of subcutaneous extracellular matrix proteins into the circulation is a key step in hemostasis initiation after vascular injury. However, in cases of severe trauma, extracellular matrix proteins are unable to cover the wound, making it difficult to effectively initiate hemostasis and resulting in a series of bleeding events. Acellular-treated extracellular matrix (ECM) hydrogels are widely used in regenerative medicine and can effectively promote tissue repair due to their high mimic nature and excellent biocompatibility. ECM hydrogels contain high concentrations of extracellular matrix proteins, including collagen, fibronectin, and laminin, which can simulate subcutaneous extracellular matrix components and participate in the hemostatic process. Therefore, it has unique advantages as a hemostatic material. This paper first reviewed the preparation, composition and structure of extracellular hydrogels, as well as their mechanical properties and safety, and then analyzed the hemostatic mechanism of the hydrogels to provide a reference for the application and research, and development of ECM hydrogels in the field of hemostasis.

Bactericidal Mechanisms of Chlorine Dioxide against Beta-Hemolytic Streptococcus CMCC 32210

Chlorine dioxide is a globally recognized green and efficient disinfectant. This study aims to investigate the bactericidal mechanism of chlorine dioxide using beta-hemolytic Streptococcus (BHS) CMCC 32210 as a representative strain. BHS was exposed to chlorine dioxide, the minimum bactericidal concentration (MBC) values of chlorine dioxide against BHS were determined by the checkerboard method in preparation for subsequent tests. Cell morphology was observed using electron microscopy. Protein content leakage, adenosine triphosphatase (ATPase) activity, and lipid peroxidation were determined by kits, and DNA damage was determined using agar gel electrophoresis. The concentration of chlorine dioxide during disinfection showed a linear relationship with the concentration of BHS. Scanning electron microscopy (SEM) results showed that chlorine dioxide caused significant damage to the cell walls of BHS at a concentration of 50 mg/L, but had no significant effect on Streptococcus exposed to different exposure times. Furthermore, the extracellular protein concentration increased with increasing chlorine dioxide concentration, while the total protein content remained unchanged. The activities of Na+/K+-ATPase and Ca2+/Mg2+-ATPase decreased with increasing chlorine dioxide concentration. Chlorine dioxide treatment led to significant lipid peroxidation and DNA degradation in BHS. Leakage of intracellular components indicated that chlorine dioxide damaged the cell membrane of BHS. Chlorine dioxide exposure resulted in oxidative damage to lipids and proteins, which negatively impacted the cell wall and membrane of Streptococcus. This caused increased permeability and inactivation of key enzymes (Na+/K+-ATPase and Ca2+/Mg2+-ATPase) involved in respiratory metabolism, ultimately leading to DNA degradation and bacterial death due to either content leakage or metabolic failure.

Extracellular vesicles in systemic juvenile idiopathic arthritis.

Systemic juvenile idiopathic arthritis is a chronic pediatric inflammatory disease of unknown etiology, characterized by fever, rash, hepatosplenomegaly, serositis, and arthritis. We hypothesized that intercellular communication, mediated by extracellular vesicles, contributes to systemic juvenile idiopathic arthritis pathogenesis and that the number and cellular sources of extracellular vesicles would differ between inactive and active states of systemic juvenile idiopathic arthritis and healthy controls. We evaluated plasma from healthy pediatric controls and patients with systemic juvenile idiopathic arthritis with active systemic flare or inactive disease. We isolated extracellular vesicles by size exclusion chromatography and determined total extracellular vesicle abundance and size distribution using microfluidic resistive pulse sensing. Cell-specific extracellular vesicle subpopulations were measured by nanoscale flow cytometry. Isolated extracellular vesicles were validated using a variety of ways, including nanotracking and cryo-electron microscopy. Extracellular vesicle protein content was analyzed in pooled samples using mass spectrometry. Total extracellular vesicle concentration did not significantly differ between controls and patients with systemic juvenile idiopathic arthritis. Extracellular vesicles with diameters <200 nm were the most abundant, including the majority of cell-specific extracellular vesicle subpopulations. Patients with systemic juvenile idiopathic arthritis had significantly higher levels of extracellular vesicles from activated platelets, intermediate monocytes, and chronically activated endothelial cells, with the latter significantly more elevated in active systemic juvenile idiopathic arthritis relative to inactive disease and controls. Protein analysis of isolated extracellular vesicles from active patients showed a proinflammatory profile, uniquely expressing heat shock protein 47, a stress-inducible protein. Our findings indicate that multiple cell types contribute to altered extracellular vesicle profiles in systemic juvenile idiopathic arthritis. The extracellular vesicle differences between systemic juvenile idiopathic arthritis disease states and healthy controls implicate extracellular vesicle-mediated cellular crosstalk as a potential driver of systemic juvenile idiopathic arthritis disease activity.

Effects of 4-tert-butylpyrocatechol and tea polyphenol on growth, physiology and antioxidant responses in Microcystis aeruginosa

Global warming has increased the frequency of Microcystis aeruginosa blooms, leading to the deterioration of water quality and loss of biodiversity. Therefore, developing effective strategies for controlling M. aeruginosa blooms has become an important research topic. Plant extracts, 4‑tert-butylpyrocatechol (TBC) and tea polyphenol (TP) are commonly used for water purification and to increase fish immunity, which have great potential to inhibit cyanobacterial blooms. The inhibitory effects of TBC and TP on M. aeruginosa were investigated in terms of growth characteristics, cell membrane morphology, physiological, photosynthetic activities, and antioxidant enzymes activities. The results showed that TBC and TP inhibited the growth of M. aeruginosa by decreasing the chlorophyll fluorescence transients or increasing the antioxidant enzymes activities of M. aeruginosa. TBC damaged the cell morphology of M. aeruginosa, reduced extracellular polysaccharides and protein contents, and up-regulated the antioxidant activity-related gene (sod and gsh) expressions of M. aeruginosa. TP significantly decreased the photosynthetic pigment content, influenced the phycobiliprotein content, and strongly down-regulated the photosynthesis-related gene (psbA, psaB, and rbcL) relative expressions of M. aeruginosa. TBC caused significant oxidative stress, dysfunction of physiological metabolic processes, and damaged crucial biomacromolecules (e.g., lipids, proteins and polysaccharides), prompted the loss of cell integrity, ultimately leading to the death of M. aeruginosa. However, TP depressed photosynthetic activities and consequently inhibited the transfer of electrons, affected the electron transfer chain, decreased the photosynthetic efficiency, and eventually caused the death of M. aeruginosa cells. Our study showed the inhibitory effects and algicidal mechanisms of TBC and TP on M. aeruginosa, and provide a theoretical basis for restrain the overgrowth of M. aeruginosa.

Phosphorus level impacts luteolin effect on Microcystis aeruginosa growth and microcystin-pollution risk − Novel perspective from correlation between exopolymers substances fractions and microcystin-production/release

Luteolin as a phytogenic algicide can inhibit the growth and microcystins (MCs) release of Microcystis, a dominant genus during cyanobacterial blooms, but how phosphorus (P) level impacts luteolin effect on its growth and MC-pollution risk is unclear. By employing Microcystis aeruginosa as test alga, this study addressed this concern and explored response mechanisms from novel insights of relationship between extracellular polysaccharide (ex-poly) and protein (ex-pro) contents and MC-production/release. At each P level (0.05–5 mg/L), rising luteolin dose more greatly inhibited Microcystis growth and MC-pollution risk, with growth inhibition ratio of around 10%–30%, 20%–50% and 40%–90% for 3, 6 and 12 mg/L luteolin, respectively, but almost increasingly enhanced cellular ability of MC-production/conservation and total and bound ex-poly/ex-pro production. Rising P level promoted Microcystis growth and intracellular/extracellular MCs content (IMC, EMC) in test system at each luteolin dose, thus higher P level weakened algicidal and MC-removal effects of luteolin, indicating that P-decrease was required for stronger application outcome of luteolin. Total and bound ex-poly/ex-pro amount were positively correlated with cellular MC-production/conservation ability, IMC and EMC, which constituted cooperative stress-defense of Microcystis at each P level. Besides, rising luteolin dose posed stronger algicidal effect by inactivating gene expression involving peroxidase synthesis (especially at P-limitation), photosynthesis and P acquisition, while rising P level alleviated algicidal and MC-pollution inhibition effects of luteolin by enhancing gene expression involving N acquisition and peroxidase synthesis. This study shed novel insights for P-dependent effect and mechanisms of luteolin on toxigenic Microcystis growth and MC-pollution control, which guided to mitigating toxigenic Microcystis-dominated cyanobacterial blooms in different P-level water areas.

Composite carrier enhanced bacterial adhesion and nitrogen removal in partial nitrification/anammox process

The rapid start-up and stable operation of one-stage (Partial nitrification/anammox) PN/A process for low-ammonium wastewater are difficult to be achieved, and many carriers are designed to solve this problem. Here, a composite carrier was developed, in which sepiolite and non-woven fabrics were assembled in polypropylene spherical shells. At the start-up phase, PA reactor using the composite carriers reached a higher nitrogen removal rate of 134.50 ± 19.60 mg·N·L−1d−1, in contrast to that of 48.85 ± 19.64 mg·N·L−1d−1 in the PB reactor without sepiolite carriers. When the final influent ammonium concentration of PN/A process is 100 mg/L, the total nitrogen removal efficiency can reach 72 ± 0.03 %. High biomass immobilization ability of composite carrier was evidenced by the greater adsorption trend between sludge and sepiolite than that between sludge and non-woven fabrics, where hydrophobic interaction and Van der Waals interaction played a major role. Extracellular protein (PN) content and the ratio of PN and extracellular polysaccharide of samples in PA were significantly higher than those in PB, verifying higher biofilm formation ability on the composite carrier. The composite carrier also increased the abundance of dominant bacteria in PN/A process, especially AOB, the relative abundance of which reached 46.11 %. And it increased the abundance of essential functional genes for nitrogen conversion as their perfect acid neutralizing effects. This study is of great significance in improving the start-up speed and stable operation of this process.