Preparing high-performance microspheres based on the chitosan-assisted dispersion of reduced graphene oxide in aqueous solution for bilirubin removal.

The removal of excess bilirubin from blood is of great clinical importance. Reduced graphene oxide (rGO) is often used to efficiently remove bilirubin. However, thin rGO pieces tend to aggregate in the aqueous phase because they are hydrophobic. In this context, we propose an effective strategy based on the chitosan-assisted (CS-assisted) dispersion of rGO to produce high-performance bilirubin-adsorbing microspheres. CS possesses a hydrophobic CH structure, which offers strong hydrophobic interactions with rGO that assist its dispersion, and the large number of hydrophilic sites of CS increases the hydrophilicity of rGO. CS serves as a dispersant in a surfactant-like manner to achieve a homogeneous and stable CS/rGO dispersion by simply and gently stirring CS and rGO in a LiOH/KOH/urea/H2O system. Subsequently, CS/rGO hybrid microspheres were prepared by emulsification. CS ensures blood compatibility as a base material, and the entrapped rGO contributes to mechanical strength and a high adsorption capacity. The CS/rGO microspheres exhibited a high bilirubin adsorption capacity (215.56 mg/g), which is significantly higher than those of the rGO and CS microspheres. The determined mass-transfer factors revealed that the rich pores of the CS/rGO microspheres promote mass transfer during bilirubin adsorption (equilibrium is almost achieved within 30 min). The CS/rGO microspheres are promising candidates for bilirubin removal owing to a combination of high strength, blood compatibility, and high adsorption capacity.

Tanreqing suppresses the proliferation and migration of non-small cell lung cancer cells by mediating the inactivation of the HIF1α signaling pathway via exosomal circ-WDR78.

Circular RNAs (circRNAs) have emerged as regulators of cancer progression, including non-small cell lung cancer (NSCLC). Tanreqing (TRQ), a traditional Chinese medicine, is used clinically for respiratory diseases. RT-qPCR quantified circ-WDR78 expression in NSCLC cells. Cell growth, apoptosis, invasion, and migration were assessed by functional assays. RNA-binding protein immunoprecipitation (RIP), luciferase reporter, and RNA pull-down assays determined the competing endogenous RNA (ceRNA) network of circ-WDR78. The interaction between HIF1α and CD274 (PD-L1) promoter was analyzed by chromatin immunoprecipitation (ChIP). Circ-WDR78 expression was up-regulated in TRQ-treated NSCLC cells. Functionally, circ-WDR78 exhibited anti-tumor effects in these cells. Additionally, circ-WDR78 could also induce reactive oxygen species (ROS) accumulation by down-regulating HIF1α expression, promoting autophagy. Mechanistically, circ-WDR78 destabilizes HIF1α via the miR-1265/FBXW8 axis. TRQ-induced exosome secretion from NSCLC cells inhibits PD-L1 expression, preventing immune escape. We found that TRQ-treated NSCLC cells secrete exosomes to transmit circ-WDR78 to untreated NSCLC cells, inhibiting the malignancy of recipient tumor cells. In conclusion, TRQ inhibits NSCLC cell proliferation, invasion, and migration through exosomal circ-WDR78-mediated inactivation of the HIF1α signaling pathway, providing potential insight into TRQ injection for NSCLC treatment.Communicated by Ramaswamy H. Sarma.

Rigid crosslink improves the surface area and porosity of ß-cyclodextrin beads for enhanced adsorption of flavonoids.

Many reported ß-cyclodextrin (ß-CD) polymers have poor flavonoid adsorption performance due to their low surface area and porosity resulting from the compact stack of the ß-CD molecules crosslinked by flexible crosslinkers. Here, we propose a rigid crosslink strategy that uses phytic acid (PA) having rigid cyclic group as crosslinkers, achieving a high-surface-area (61.42-140.23 m2/g) and porous ß-CD beads. The improved surface area and porosity are attributed to the rigid cyclic groups in PA, which expand the network structure of ß-CD polymers. Benefitting from the advantages, the optimized PA-crosslinked ß-CD (PA-ß-CD) beads have an over tenfold increased adsorption amount and an threefold increased diffusivity for rutin compared with traditional non-porous ß-CD beads crosslinked by epichlorohydrin. Moreover, dynamic adsorption experiments reveal that PA-ß-CD beads are able to treat about 1100 mL of rutin solution (0.05 mg/mL), over 5 times higher than that of the non-porous ß-CD beads (200 mL). These results demonstrate the promise of PA-ß-CD beads for rapid and high-capacity adsorption of rutin.

Utilization of an aqueous two-phase emulsification to prepare bimodal porous cellulose monolith for efficient adsorption of bovine serum albumin.

Cellulose monolith has garnered significant interest in the field of biochromatography, which lies in its interconnected porous structure, large surface area and biocompatibility. In this context, we propose a novel approach for preparing cellulose monoliths using an aqueous two-phase system devoid of any organic solvents and surfactants. In this strategy, emulsifying cellulose solution into PEG 20,000 solution gives bicontinuous aqueous phases and further porous cellulose monolith after regeneration of dissolved cellulose. And the macroporous channels are derived from the removal of the PEG 20,000 aqueous phase while the micropores are from the phase separation of the cellulose phase. Physical characterizations reveal the obtained cellulose monolith exhibits exceptional column permeability of 1.36 × 10-11 m2 and a substantial surface area of 39.34 m2/g. Furthermore, cellulose monolith is functionalized with diethyl ethylamine hydrochloride (DEAE-HCl) to evaluate its potential as an anion adsorbent. Experimental results reveal that the DEAE-modified cellulose monolith possesses of adsorptive capacity of 316.58 mg/g of bovine serum albumin, along with fast adsorption kinetic. This study introduces an innovative strategy for fabricating porous cellulose monoliths tailored for biochromatography applications.

Humic acid-dependent respiratory growth of Methanosarcina acetivorans involves pyrroloquinoline quinone.

Although microbial humus respiration plays a critical role in organic matter decomposition and biogeochemical cycling of elements in diverse anoxic environments, the role of methane-producing species (methanogens) is not well defined. Here we report that a major fraction of humus, humic acid reduction enhanced the growth of Methanosarcina acetivorans above that attributed to methanogenesis when utilizing the energy sources methanol or acetate, results which showed both respiratory and fermentative modes of energy conservation. Growth characteristics with methanol were the same for an identically cultured mutant deleted for the gene encoding a multi-heme cytochrome c (MmcA), results indicating MmcA is not essential for respiratory electron transport to humic acid. Transcriptomic analyses revealed that growth with humic acid promoted the upregulation of genes annotated as cell surface pyrroloquinoline quinone (PQQ)-binding proteins. Furthermore, PQQ isolated from the membrane fraction was more abundant in humic acid-respiring cells, and the addition of PQQ improved efficiency of the extracellular electron transport. Given that the PQQ-binding proteins are widely distributed in methanogens, the findings extend current understanding of microbial humus respiration in the context of global methane dynamics.

Respiration-driven methanotrophic growth of diverse marine methanogens.

Anaerobic marine environments are the third largest producer of the greenhouse gas methane. The release to the atmosphere is prevented by anaerobic 'methanotrophic archaea (ANME) dependent on a symbiotic association with sulfate-reducing bacteria or direct reduction of metal oxides. Metagenomic analyses of ANME are consistent with a reverse methanogenesis pathway, although no wild-type isolates have been available for validation and biochemical investigation. Herein is reported the characterization of methanotrophic growth for the diverse marine methanogens Methanosarcina acetivorans C2A and Methanococcoides orientis sp. nov. Growth was dependent on reduction of either ferrihydrite or humic acids revealing a respiratory mode of energy conservation. Acetate and/or formate were end products. Reversal of the well-characterized methanogenic pathways is remarkably like the consensus pathways for uncultured ANME based on extensive metagenomic analyses.

Application of cellulose to chromatographic media: Cellulose dissolution, and media fabrication and derivatization.

As the cornerstone of chromatographic technology, the development of high-performance chromatographic media is a crucial means to enhance the purification efficiency of biological macromolecules. Cellulose is a popular biological separation medium due to its abundant hydroxyl group on the surface, easy modification and, weak non-specific adsorption. In this paper, the development of cellulosic solvent systems, typical preparation methods of cellulosic chromatographic media, and the enhancement of chromatographic properties of cellulosic chromatographic media by polymeric ligand grafting strategies and their mechanism of action are reviewed. Ultimately, based on the current research status, a promising outlook for the preparation of high-performance cellulose-based chromatographic media was presented.

Multi-size optimization of macroporous cellulose beads as protein anion exchangers: Effects of macropore size, protein size, and ligand length.

Multi-size optimization of ion exchangers based on protein characteristics and understanding of underlying mechanism is crucial to achieve maximum separation performance in terms of adsorption capacity and uptake kinetic. Herein, we characterize the effects of three different sizes, macropore size, protein size, and ligand length, on the protein adsorption capacity and uptake kinetic of macroporous cellulose beads, and provide insights into the underlying mechanism. In detail, (1) for smaller bovine serum albumin, macropore size has a negligible effect on the adsorption capacity, while for larger γ-globulin, larger macropores improve the adsorption capacity due to the high accessibility of binding sites; (2) there is a critical pore size (CPZ), at which the adsorption uptake kinetic is minimum. When pore sizes are higher than the CPZ, uptake kinetics are enhanced by pore diffusion. When pore sizes are lower than CPZ, uptake kinetics are enhanced by surface diffusion; (3) increasing ligand length improves the adsorption capacity by three-dimensionally extended polymer chains in pores and enhances uptake kinetic by improved surface diffusion. This study offers an integrated perspective to qualitatively assess the effects of multiple sizes, providing guidance for designing advanced ion exchangers for protein chromatography.

Medium-long term prognosis prediction for idiopathic pulmonary fibrosis patients based on quantitative analysis of fibrotic lung volume.

PURPOSE: To investigate the prognostic value of quantitative analysis of CT among patients with idiopathic pulmonary fibrosis (IPF) by quantifying the fibrosis extent and to attempt to provide precise medium-long term prognostic predictions for individual patients. METHODS: This was a retrospective cohort study that included 95 IPF patients in Zhongshan Hospital, Fudan University. 64 patients firstly diagnosed with IPF from 2009 to 2015 was included as the derivation cohort. Information regarding sex, age, the Gender-Age-Physiology (GAP) index, high-resolution computed tomography (HRCT) images, survival status, and pulmonary function parameters including forced vital capacity (FVC), FVC percent predicted (FVC%pred), diffusing capacity of carbon monoxide (DLCO), DLCO percent predicted (DLCO%pred), carbon monoxide transfer coefficient (KCO), KCO percent predicted (KCO%pred) were collected. 31 patients were included in the validation cohort. The Synapse 3D software was used to quantify the fibrotic lung volume (FLV) and total lung volume (TLV). The ratio of FLV to TLV was calculated and labeled CTFLV/TLV%, reflecting the extent of fibrosis. All the physiological variants and CTFLV/TLV% were analyzed for the dimension of survival through both univariate analysis and multivariate analysis. Formulas for predicting the probability of death based on the baseline CTFLV/TLV% were calculated by logistic regression, and validated by the validation cohort. RESULTS: The univariate analysis indicated that CTFLV/TLV% along with DLCO%pred, KCO%pred and GAP index were significantly correlated with survival. However, only CTFLV/TLV% was meaningful in the multivariate analysis for prognostic prediction (HR 1.114, 95% CI 1.047-1.184, P = 0.0006), and the best cutoff was 11%, based on receiver operating characteristic (ROC) curve analysis. The survival times for the CTFLV/TLV% ≤ 11% and CTFLV/TLV% > 11% groups were significantly different. Given the CTFLV/TLV% data, the death probability of a patient at 1 year, 3 years and 5 years could be calculated by using a particular formula. The formulas were tested by the validation cohort, showed high sensitivity (88.2%), specificity (92.8%) and accuracy (90.3%). CONCLUSION: Quantitative volume analysis of CT might be useful for evaluating the extent of fibrosis in the lung. The CTFLV/TLV% could be a valuable biomarker for precisely predicting the medium-long term prognosis of individual patients with IPF.

Boronate-immobilized cellulose nanofiber-reinforced cellulose microspheres for pH-dependent adsorption of glycoproteins.

High strength and excellent selectivity are two important aspects of porous cellulose microspheres as adsorbents for protein separation. For this purpose, self-reinforced all-cellulose microspheres (SCMs) with high strength were fabricated using natural cellulose nanofibers (CNFs) as fillers and then immobilized via 3-aminophenylboronic acids as affinity ligands for selective enrichment of glycoproteins. In particular, the inherent stiffness of entrapped CNFs endowed SCMs with more inflexibility, because the stress can be efficiently transferred from the network of SCMs to the stiff CNFs during the separation process. Besides, SCMs, as an all-cellulose material with homogenous surface chemistry and pore structure characteristics, are more suitable as supports for adsorbents. Finally, the SCMs were immobilized with 3-aminophenylboronic acids (BA/EPI-SCMs) and tested their performance in affinity adsorption of glycoproteins. BA/EPI-SCMs showed fast separation, high adsorption amount, and excellent selectivity toward glycoproteins.

Preparation of cellulose-based chromatographic medium for biological separation: A review.

Cellulose is a kind of renewable linear polysaccharide with good safety, hydrophilicity, biocompatibility and biodegradability and has become a commonly used chromatographic medium for biological separation and purification. The purpose of this paper is to describe the recent developments in the applications of cellulose-based absorbents as chromatographic medium. This review also attempts to explain the current situation of cellulose-based chromatographic medium from the aspects of cellulose dissolution, common strategies for generating spherical particles of cellulose and its derivatives (such as titration, emulsification, microfluidic and other synthesis methods) and improvement of adsorption properties. Furthermore, non-spherical cellulose-based chromatographic medium are also described briefly.

Proteinaceous porous nanofiber membrane-type adsorbent derived from amyloid lysozyme protofilaments for highly efficient lead(II) biologic scavenging.

To overcome the technical bottleneck of fine amyloid lysozyme fibrils in environmental engineering, a novel co-operative strategy was identified to fabricate free-standing lysozyme complex nanofibers based membrane-type adsorbent (Lys-CNFs membrane) through a combination of vacuum filtration for lead remediation. The composition of the membrane integrated the linear amyloid protofilaments that were obtained by acid-heating fibrillation and polydopamine that adjusted the fibers' diameters and surface chemistry. As expected, the Lys-CNFs membrane not only showed nanofibrous morphology and layer stacking architecture but presented a hierarchical macro-mesoporous structure along with a high surface area of 220.4 m2/g. Besides, the thermal stability up to 200 â„ƒ and wetting nature of below 2 s endowed its further applicability. Adsorption experiments showed that Lys-CNFs membrane can effectively uptake Pb(II) ions with acceptable selectivity, high adsorption capacity of 270.3 mg/g, rapid equilibrium kinetic within only 10 mins, and good reusability that dropped by 14.9% efficiency even after five cycles, indicating that Lys-CNFs membrane can be as an affordable technology for alleviating the lead pollution issues.

Macroporous chitin microspheres prepared by surfactant micelle swelling strategy for rapid capture of lead (II) from wastewater.

Heavy metal pollution of water source continues to be one of the most serious environmental problems which have attracted major global concern. Here, a macroporous chitin microsphere is prepared by surfactant micelle swelling strategy followed by modification with tetraethylenepentamine for Pb2+ removal from wastewater. The resultant adsorbent not only exhibits fast adsorption kinetic (>80% of its equilibrium uptake within 20 min) but also has high adsorption capacity of 218.4 ± 6.59 mg/g and excellent reusability (>75% of its initial adsorption capacity after five adsorption/desorption cycles). More importantly, under the continuous operating mode, the adsorbent can treat about 39,000 kg water/kg adsorbent, and the Pb2+ concentration decreases from 2000 µg/L to smaller than 10 µg/L, meeting the drinking water standard recommended by the World Health Organization (10 µg/L). All results indicate that the tetraethylenepentamine-modified macroporous chitin microspheres have great potential in the treatment of heavy metal contamination.

Bisphosphonated-immobilized porous cellulose monolith with tentacle grafting by atom transfer radical polymerization for selective adsorption of lysozyme.

Here, a m-xylene bisphosphonate immobilized tentacle-type cellulose monolith (BP-PCM) is prepared by atom transfer radical polymerization for lysozyme purification. In the preparation, the m-xylene bisphosphonate was anchored glycidyl methacrylate and then polymerized to enhance the flexibility of the ligands to improve lysozyme adsorption capacity, and glycerol monomethacrylate serves as spacer to further optimize the layers structure and ligands density of the grafted tentacles for satisfactory adsorption capacity. The maximum static and dynamic adsorption capacity (10% breakthrough) of BP-PCM reach to 169.6 and 102.6 mg mL-1, respectively. Moreover, BP-PCM displays weak nonspecific adsorption and is able to successfully enrich lysozyme from diluted chicken egg white, indicating the excellent selectivity. The results demonstrated that BP-PCM is promising for use as high-capacity protein chromatography.

Tentacle-type poly(hydroxamic acid)-modified macroporous cellulose beads: Synthesis, characterization, and application for heavy metal ions adsorption.

Herein, a facile yet efficient template method to fabricate macroporous cellulose beads (MCBs) is reported. In this method, micro-size CaCO3 is utilized to create macroporous structure for fast mass transfer, and tentacle-type poly(hydroxamic acid) as adsorption ligand is immobilized on the MCBs to improve adsorption capacity. The obtained tentacle-type poly(hydroxamic acid)-modified MCMs (TP-CMCBs) show uniform spherical shape (about 80 µm), bimodal pore system (macropores≈3.0 µm; diffusional pores≈14.5 nm), and high specific surface area (52.7 m2/g). The adsorption performance of TP-CMCBs is evaluated by heavy metal ions adsorption. TP-CMCBs exhibit not only high adsorption capacities (342.5, 261.5 and 243.2 mg/g for Cu2+, Mn2+ and Ni2+, respectively.), but also fast adsorption rate (>70% of its equilibrium uptake within 30 min). Additionally, TP-CMCBs have excellent reusability, as evidenced by that the adsorption capacities have no obvious change even after five-time consecutive adsorption-desorption cycles. All results demonstrate that the proposed TP-CMCBs have great potential in removal of heavy metal ions.

Preparation and characterization of highly porous cellulose-agarose composite chromatographic microspheres for enhanced selective separation of histidine-rich proteins.

In this work, the porous cellulose-agarose microspheres with high specific surface area and enhanced mechanical strength are prepared by a novel chemical crosslinking method. The crosslinking reaction homogeneously proceeds between polysaccharides, and the covalent bonding network is generated to replace the inherent hydrogen bonding network of cellulose. The prepared microspheres exhibit low crystallinity of 12.45%, which means high content of amorphous regions. The micro-meso-macroporous structure of microspheres in morphology is conducive to high permeability and adsorption capacity, and the microspheres possess high specific surface area of 183.81 m2/g. The affinity chromatographic microspheres are prepared by immobilizing Cu2+, which exhibits high adsorption capacity of 197.65 mg/g for bovine hemoglobin (BHb), fast adsorption rate wihin 40 minutes, well-selectivity, and excellent recyclability in ten cycles. We expect that this work to provide an outstanding candidate for the high performance of biomacromolecular purification.

Direct preparation of porous cellulose microspheres via a self-growth process on bamboo fibers and their functionalization for specific adsorption of histidine-rich proteins.

The traditional preparation of cellulose microspheres always involves tedious synthetic procedures (e.g., dissolution, emulsification and regeneration) and inevitable organic solvents, which undergoes both high production cost and environmental contamination. To overcome these issues, a feasible and green synthesis strategy is proposed to construct porous cellulose microspheres (PCMs) via one-step spontaneous formation relying on sodium periodate oxidation of pure bamboo fibers. By this strategy, a cluster of robust cellulose microspheres grow up on the surface of bamboo fibers in aqueous phase through amorphous oxidized cellulose self-assembly accumulation and then drop out when their sizes increase to about 15 µm. After being immobilized with Cu(II), the prepared cellulose microspheres serve as metal affinity adsorbent for proteins adsorption, showing high adsorption capacity, good selectivity and excellent reusability for bovine hemoglobin (BHb). Together with green and easy synthesis, the novel cellulose microspheres show a promising alternative to commercially available adsorbent support.

High-strength, blood-compatible, and high-capacity bilirubin adsorbent based on cellulose-assisted high-quality dispersion of carbon nanotubes.

Excess bilirubin can accumulate in body organs and has serious effects on human health. In this work, a simple engineering strategy, based on cellulose-assisted high-quality dispersion of carbon nanotubes (CNTs), is proposed to produce high-performance bilirubin adsorbents. By dispersing cellulose and CNTs in NaOH/thiourea aqueous solution, a homogeneous and stable cellulose/CNTs solution is achieved. The obtained cellulose/CNTs solution is applied for the fabrication of cellulose/CNTs microspheres (CCMs), in which cellulose serves as a base material and guarantees the blood compatibility of the composite material, and CNTs contribute to the improved mechanical strength and high adsorption capacity. To further improve blood compatibility and adsorption capacity, lysine is immobilized on the CCMs. The obtained lysine-modified CCMs (LCCMs) exhibit a large surface area (171.31 m2/g) and hierarchically porous structure. Experimental results demonstrate LCCMs have high bilirubin adsorption capacity (204.12 mg/g) that is significantly higher than most of the reported adsorbents. The combination of high strength, blood compatibility, and high adsorption capacity positions the LCCMs as a promising candidate for bilirubin removal.

Simultaneous determination of sulfonamides in milk: In-situ magnetic ionic liquid dispersive liquid-liquid microextraction coupled with HPLC.

Four novel organic magnetic ionic liquids were synthesized and characterized. A new method of in-situ magnetic ionic liquid dispersive liquid-liquid microextraction coupled with HPLC was established to simultaneously separate, preconcentrate and determine trace amount of sulfonamides in milk samples for the first time. In this work, extraction procedure was free of volatile organic solvent. Extraction equilibrium was immediately achieved without further operation such as vortex shaking. Magnetic separation of two phases was effectively achieved by applying external magnetic field without need of centrifugation. Furthermore, only a very small amount of inorganic salts were generated. This is the first time to realize all these advantages in an analytical method. Under the optimal conditions, satisfactory enrichment factors, precisions, linear range and low limit of detection and quantitation were acquired. The validated method was successfully applied in the simultaneous analysis of 5 sulfonamides in real milk samples.

The C-H activated controlled mono- and di-olefination of arenes in ionic liquids at room temperature.

In this study, controlled mono and di-olefination of arenes was first realized at room temperature via the C-H bond activation in ionic liquids, probably due to the positive effects of ionic liquids. It is an energy-saving routes in industrial production without the need for heating equipment. Different catalysts were screened, and it was found that [Ru(p-cymene)Cl2]2 generated mono-olefinated products predominantly while [Cp*RhCl2]2 selectively gave di-olefinated products. These catalysts ([BMIM]NTf2 and [BMIM]PF6) as green and recyclable reaction media are highly efficient under mild conditions. This reaction process can avoid any volatile and environmentally toxic organic solvents, and is much safer without the need for pressure-tight equipment. A wide substrate scope with good yields and satisfactory selectivity was achieved. The reactions can be scaled up to gram-scale. Furthermore, an expensive rhodium/ruthenium catalytic system was recycled for at least 6 times with consistently high catalytic activity, which was economical and environmental friendly from an industrial point of view. According to the mechanistic study, the C-H bond cleavage was probably achieved via the concerted metalation-deprotonation. This technique can be applied in the synthesis of various valuable unsaturated aromatic compounds and shows a great potential for industrial production.