Identification of a distal enhancer that determines the expression pattern of acute phase marker C-reactive protein.

C-reactive protein (CRP) is a major acute phase protein and inflammatory marker, the expression of which is largely liver specific and highly inducible. Enhancers are regulatory elements critical for the precise activation of gene expression, yet the contributions of enhancers to the expression pattern of CRP have not been well defined. Here, we identify a constitutively active enhancer (E1) located 37.7 kb upstream of the promoter of human CRP in hepatocytes. By using chromatin immunoprecipitation, luciferase reporter assay, in situ genetic manipulation, CRISPRi, and CRISPRa, we show that E1 is enriched in binding sites for transcription factors STAT3 and C/EBP-ß and is essential for the full induction of human CRP during the acute phase. Moreover, we demonstrate that E1 orchestrates with the promoter of CRP to determine its varied expression across tissues and species through surveying activities of E1-promoter hybrids and the associated epigenetic modifications. These results thus suggest an intriguing mode of molecular evolution wherein expression-changing mutations in distal regulatory elements initiate subsequent functional selection involving coupling among distal/proximal regulatory mutations and activity-changing coding mutations.

Easily scale 3D conductive gradient fiber membrane for contaminants removal and fouling mitigation under electrochemical assistance.

An electrochemical membrane filtration system provides an innovative approach to enhance contaminant removal and mitigate membrane fouling. There is an urgent need to develop portable, versatile, and efficient electrochemical membranes for affordable wastewater treatment. Here, a 3D conductive gradient fiber membrane (CC/PVDF) with a gradient porous structure was prepared using a two-step phase inversion method. Methyl orange (MO) was utilized as model organic substance to investigate the electrochemical performance of the CC/PVDF membrane. At applied potentials of +2 V, +3 V, -2 V and -3 V, the removal efficiency of MO was 5.1, 5.3, 4.8, and 5.1 times higher than at 0 V. A dramatic flux loss of 35.02% occurred on the membrane without electrochemistry, interestingly, whereas the flux losses were only 23.59%-10.24% in the applied potential after 30 min of filtration, which were approximately 1.18, 1.28, 1.29 and 1.38 times as high as that without electrochemistry, respectively. The enhanced removal and anti-fouling performances of the membranes were attributed to the functions of electrochemical degradation, electrostatic repulsion, and electrically enhanced wettability. Electrochemical generation of Hydrogen peroxide, along with HO• radicals, was detected and direct electron transfer and HO• were proved to be the dominant oxidants responsible for MO degradation. The intermediate oxidation products were identified by mass spectrometry, and an electrochemical degradation pathway of MO was proposed based on bond-breaking oxidation, ring-opening reactions, and complete oxidation. All the findings emphasize that the ECMF system possesses superior efficiency and creative potential for water purification applications.

Polyethylene glycol-polyvinylidene fluoride/TiO2 nanocomposite polymer coatings with efficient antifouling strategies: Hydrophilized defensive surface and stable capacitive deionization.

Capacitive deionization (CDI) is flourishing as an energy-efficient and cost-effective water desalination method. However, challenges such as electrode degradation and fouling have hindered the practical deployment of CDI technology. To address these challenges, the key point of our strategy is applying a hydrophilic coating composed of polyethylene glycol (PEG)-functionalized nano-TiO2/polyvinylidene fluoride (PVDF) to the electrode interface (labeled as APPT electrode). The PEG/PVDF/TiO2 layer not only mitigates the co-ion depletion, but also imparts the activated carbon (AC) electrode hydrophilicity. As anticipated, the APPT electrode possessed an enhanced desalination capacity of 83.54 µmol g-1 and a low energy consumption of 17.99 Wh m-3 in 10 mM sodium chloride solution compared with the bare AC electrode. Notably, the APPT maintained about 93.19 % of its desalination capacity after 50 consecutive adsorption-desorption cycles in the presence of bovine serum albumin (BSA). During the trial, moreover, no obvious overall performance decline was noted in concentration reduction (Δc), water recovery (WR) and productivity (P) over 50 cycles. This strategy realizes energy-efficient, antifouling and stable brackish water desalination and has great promise for practical applications.

C-Reactive Protein Is Not the Driver Factor in Ulcerative Colitis.

Purpose: C-reactive protein (CRP) functions as a nonspecific marker in various inflammatory disorders, particularly in evaluating the efficacy of pharmacological treatments in patients with ulcerative colitis. The existing body of evidence does not offer adequate support for the direct implication of CRP in modulating the advancement of ulcerative colitis. Methods: Our study employed a rigorous mouse model. An ulcerative colitis mouse model was established by subjecting CRP-deficient mice to dextran sulfate sodium (DSS) treatment. The phenotype of the mice, which encompassed parameters such as body weight, colon length, and spleen weight, was meticulously evaluated. Additionally, various physiological and biochemical indicators were assessed, including colon histopathology, expression levels of inflammatory factors, and staining of the intestinal mucus layer. Results: The absence of CRP did not significantly affect the phenotype, physiological characteristics, and biochemical indices in a mouse model of ulcerative colitis compared to mice with wild-type CRP. Additionally, eliminating intestinal bacteria flora interference through antibiotic treatment revealed that mice lacking CRP did not demonstrate any notable variations in the ulcerative colitis model. Meanwhile, the survival rate of mice lacking CRP did not exhibit a statistically significant difference compared to wild-type mice. Conclusion: The results of our study suggest that CRP may not directly mediate ulcerative colitis. Instead, it is more likely to be a bystander that is present alongside with elevated inflammatory factors. Further investigation is warranted to determine the precise role of CRP in humans, given the significant limitations associated with the use of mouse models.

Establishing a high-performance anti-fouling PEI-ZIF-PAA membrane with improved Lewis acid-base interactions and hydrophilicity.

Membrane fouling and the trade-off between membrane permeability and selectivity restrict the potential applications of membrane filtration for water treatment. ZIF-8 was found having great permeability and antibiofouling performance, but with issue on particle aggregation makes it difficult to achieve high ZIFs loading and fabricate a defect-free molecular sieving membrane in previous research. In this study, we formed a scalable antibiofouling surface with improved permeability and fouling resistance on a PEI-ZIF-PAA membrane using a layer-by-layer assembly technique. The synergistic effects of being sandwiched between two different polyelectrolyte layers with opposite charges endowed the ZIF nanoparticles with improved stability and scalability for membrane modification. The PEI-ZIF-PAA membrane exhibited a satisfactory water flux of 120.78 LMH, which was 46.97% higher than that of the pristine PES membrane. The normalized water flux loss was serious in the absence of ZIF-8, and the flux increased with the ZIF-8 concentration. Antifouling tests suggested that the PEI-ZIF-PAA membrane possessed good antifouling performance due to the much higher surface hydrophilicity and positive Lewis acid-base interactions with foulants. The HA rejection increased with the ZIF-8 concentration and reached a maximum of 92.1% in the presence of 1.00% (w/v) ZIF-8. The membrane regeneration was tested under physical and chemical cleaning with flux recovery rates of about 85% and 95%. XDLVO analysis showed that the total interaction energy between HA and the PEI-ZIF-8-PAA membrane was 26.45 mJ/m2, and the superior antifouling performance was mainly attributed to Lewis acid-base interactions. This study indicates that ZIF-8 nanocrystals are promising materials for fabricating novel membranes for sewage treatment.

Robust self-cleaning membrane with superhydrophilicity and underwater superoleophobicity for oil-in-water separation.

The discharge of oily wastewater has increased dramatically and will bring serious environmental problems. In this work, a self-cleaning and anti-fouling g-C3N4/TiO2/PVDF composite membrane was fabricated via the layer-by-layer approach. The surface of as-prepared composite membrane displayed a superhydrophilic and underwater superoleophobic behavior under irradiation with visible light. Also, upon irradiation with visible light, the fabricated g-C3N4/TiO2/PVDF composite membrane displayed enhanced permeation flux and improved oil removal efficiency as a result of the generation of hydroxyl free radicals during the photocatalytic filtration process. Significantly, irradiation with visible light remarkably improved reusability of the composite membrane by initiating photocatalytic decomposition of deposited oil foulants, which enabled removal of over 99.75% of oils, thus reaching a nearly 100% flux recovery ratio. Furthermore, the g-C3N4/TiO2/PVDF composite membrane exhibited great anti-fouling behavior in photocatalysis-assisted filtration. The mechanistic study revealed that underwater superhydrophobicity and the generation of free hydroxyl radicals jointly contributed to membrane anti-fouling. The greatest advantages of this g-C3N4/TiO2/PVDF composite membrane are that not only does it degrades the oil pollutants, but it also makes the membrane less vulnerable to fouling.

Sonocatalytic degradation of ciprofloxacin using hydrogel beads of TiO2 incorporated biochar and chitosan.

Pharmaceuticals are necessary to be removed from environment. Herein TiO2 incorporated biochar made from pyrolysis of agricultural wastes was encapsulated into chitosan to obtain a novel hydrogel beads. This hydrogel beads executed a dual role as both adsorbent and sonocatalyst, which proved to be suitable for the removal of antibiotic ciprofloxacin (CIP) from water. The results showed that adsorption of CIP followed pseudo first order kinetics model and Langmuir adsorption isotherm model, having maximum adsorption at pH 9. Whereas the degradation was more efficient at pH 6 due to greater standard potential for •OH/H2O in acidic media. The degradation was maximum at 150 W of ultrasonic power, then decreased in presence of dissimilar electrolytes and even reduced to 0 in presence of Na3PO4. Different quenchers such as benzoquinone (BQ), Triethanolamine (TEA) and isopropyl alcohol (IPA) reduced degradation efficiency (DE) and mineralization efficiency (ME). The DE was decreased from 85.23% to 81.50% (BQ), 74.27% (TEA), and 61.77% (IPA) within 25 min. The prepared sonocatalyst was capable of regeneration with DE, remaining sufficiently high (62%) even after four regeneration steps. These results indicate that titanium-biochar/chitosan hydrogel beads (TBCB) are durable and effective for long-term CIP removal.

Intra-subunit Disulfide Determines the Conversion and Structural Stability of CRP Isoforms.

C-reactive protein (CRP) is a major human acute-phase reactant that is composed of five identical subunits. CRP dissociates into subunits at inflammatory loci forming monomeric CRP (mCRP) with substantially enhanced activities, which can be further activated by reducing the intra-subunit disulfide bond. However, conformational changes underlying the activation process of CRP are less well understood. Conformational changes accompanying the conversion of CRP to mCRP with or without reduction were examined with circular dichroism spectroscopy, fluorescence spectroscopy, electron microscopy, size-exclusion chromatography, and neoepitope expression. The conversion of CRP to mCRP follows a two-stage process. In the first stage, CRP dissociates into molten globular subunits characterized by intact secondary structure elements with greatly impaired tertiary packing. In the second stage, these intermediates completely lose their native subunit conformation and assemble into high-order aggregates. The inclusion of reductant accelerates the formation of molten globular subunits in the first step and promotes the formation of more compact aggregates in the second stage. We further show a significant contribution of electrostatic interactions to the stabilization of native CRP. The conformational features of dissociated subunits and the aggregation of mCRP may have a key impact on their activities.

A Functional Genetic Variant at the C-Reactive Protein Promoter (rs3091244) Is Not Associated With Cancer Risk in a Chinese Population.

Background: The association of genetically elevated levels of circulating C-reactive protein (CRP) with cancer risk has been extensively investigated in European populations; however, there are conflicting conclusions. The tri-allelic rs3091244 is a functionally validated genetic variant, and its allelic frequencies differ significantly between European and Asian populations. Here, we examined the association of rs3091244 with cancer risk in a Chinese population. Methods: rs3091244 was genotyped by Sanger sequencing in 4,971 cancer cases and 2,485 controls. The rs1205 and rs2794521 gene variants were also genotyped using TaqMan assays in subgroups. Results: No association was detected between the genotyped CRP variants and cancer risk, with or without distinguishing cancer types, suggesting that circulating CRP is not causally involved in tumorigenesis in Chinese populations.

Acidic pH promotes oxidation-induced dissociation of C-reactive protein.

BACKGROUND: Circulating levels of the systemic inflammation marker C-reactive protein (CRP) have been associated with increased risk and poor outcomes of many diseases, such as cardiovascular events and cancer. Accumulating evidence has indicated that the conformational rearrangement of human pentameric CRP (pCRP) to monomeric CRP (mCRP) is a prerequisite for participation in the pathogenesis. Therefore, determining the mechanism of the dissociation of pCRP into pro-inflammatory mCRP under physiological/pathological circumstances has been intriguing. METHODS: The effects of oxidative and acidic stress occurring in inflammation on pCRP were examined by electrophoresis, electron microscopy, protein fluorescence, neoepitope expression and endothelial cell responses. RESULTS: Reactive oxygen species (ROS) generated by the copper-hydrogen peroxide system could rapidly induce the dissociation of CRP at mild acidic pH within four hours, but not at physiological pH of 7.4. Meanwhile, mannitol, a ROS scavenger, could not protect against dissociation, which implied that local ROS from accessible histidine residues may be crucially beneficial to the formation of mCRP in a redox-balanced microenvironment. Furthermore, mCRP generated by ROS could be reduced by DTT, which indicated the exposure of functional motif aa35-47, and showed potent proinflammatory actions on endothelial cells, comparable to mCRP generated by urea. CONCLUSION: dissociation of pCRP to mCRP could be rapidly induced by ROS from copper- hydrogen peroxide system in dependence on mildly acidic stress regardless of a redox-balanced microenvironment.