Alcohol Pretreatment to Eliminate the Interference of Micro Additive Particles in the Identification of Microplastics Using Raman Spectroscopy.

Raman spectroscopy is an indispensable tool in the analysis of microplastics smaller than 20 µm. However, due to its limitation, Raman spectroscopy may be incapable of effectively distinguishing microplastics from micro additive particles. To validate this hypothesis, we characterized and compared the Raman spectra of six typical slip additives with polyethylene and found that their hit quality index values (0.93-0.96) are much higher than the accepted threshold value (0.70) used to identify microplastics. To prevent this interference, a new protocol involving an alcohol treatment step was introduced to successfully eliminate additive particles and accurately identify microplastics. Tests using the new protocol showed that three typical plastic products (polyethylene pellets, polyethylene bottle caps, and polypropylene food containers) can simultaneously release microplastic-like additive particles and microplastics regardless of the plastic type, daily-use scenario, or service duration. Micro additive particles can also adsorb onto and modify the surfaces of microplastics in a manner that may potentially increase their health risks. This study not only reveals the hidden problem associated with the substantial interference of additive particles in microplastic detection but also provides a cost-effective method to eliminate this interference and a rigorous basis to quantify the risks associated with microplastic exposure.

Novel 2,3-Dialdehyde Cellulose-Based Films with Photodynamic Inactivation Potency by Incorporating the ß-Cyclodextrin/Curcumin Inclusion Complex.

Antibacterial packaging film mediated by photodynamic inactivation (PDI) is a new concept in food industry. The objective of this study was to fabricate a green 2,3-dialdehyde cellulose (DAC)-based antimicrobial film with PDI potency by incorporating the ß-cyclodextrin/curcumin (ß-CD/Cur) complex as a photosensitizer. The PDI-mediated films were characterized by evaluating the surface morphology, chemical structure, light transmittance, mechanical properties, photochemical and thermal stability, and water solubility. The results showed that the DAC-CD/Cur films were soluble in water and mechanically strong with a tensile strength of 63.87 MPa and an elongation break of 1.32%, which was attributed to the formation of hydrogen bonds between DAC and ß-CD/Cur molecules. Meanwhile, the composite films possessed a good light transmittance but impeded the penetration of ultraviolet light and efficiently delayed the degradation of curcumin. More importantly, the PDI-mediated films exhibited a broad-spectrum ability to kill Listeria monocytogenes, Vibrio parahaemolyticus, and Shewanella putrefaciens in pure culture. Notably, they also potently inactivated these harmful bacteria on ready-to-eat salmon with a maximum of ∼4 Log CFU/g (99.99%) reduction after 60 min irradiation (13.68 J/cm2). Therefore, the PDI-mediated DAC-CD/Cur films are novel and promising antimicrobial food packaging films in food industry.

Differential degradation rate and underlying mechanism of a collagen/chitosan complex in subcutis, spinal cord and brain tissues of rat.

Degradation rate is an important index for evaluating biomaterials. The authors' aim was to determine whether the degradation rate of biomaterials is different in distinct tissues and to clarify the underlying mechanism of degradation. The collagen-chitosan (CG-CS) composite scaffolds were prepared using freeze-drying technology. The porosity, water absorption and swelling ratio of the scaffolds were tested in vitro. The scaffolds were implanted into the subcutis, spinal cord and brain tissues of SD rats, the rate of degradation was assessed by continuous monitoring of weight loss, the pathological changes of target areas were observed by histological staining, and matrix metalloproteinase 9 (MMP-9) and lysozyme were detected at the rapid stage of degradation of the scaffolds. Physical and chemical property testing confirmed that CG-CS composite scaffold components can meet the biological requirements of in vivo transplantation. The in vivo experimental results showed that the scaffolds were completely absorbed in the subcutis at 12 days, the scaffolds in the spinal cord and brain groups exhibited progressive mass loss starting from the 3rd week, and a substantial fraction of the scaffold was degraded at 12 weeks. HE staining found that compared with the spinal cord and brain groups, macrophages and capillaries appeared earlier in the subcutis group, and the number was significantly higher (P < 0.05). Western blot analysis showed that the MMP-9 and lysozyme levels in the subcutis were higher than those in the spinal cord and brain (P < 0.05). The results of in vivo experiments demonstrated that the CG-CS scaffold has good biocompatibility and biodegradability, while the rate of degradation was significantly different between the three tissues at the same time point. Macrophage behavior and vascularization in different parts of the body may result in control over the balance of degradation and reconstruction.

Micro and nano plastics release from a single absorbable suture into simulated body fluid.

Synthetic polymers are widely used in medical devices and implants where biocompatibility and mechanical strength are key enablers of emerging technologies. One concern that has not been widely studied is the potential of their microplastics (MPs) release. Here we studied the levels of MP debris released following 8-week in vitro tests on three typical polyglycolic acid (PGA) based absorbable sutures (PGA 100, PGA 90 and PGA 75) and two nonabsorbable sutures (polypropylene-PP and polyamide-PA) in simulated body fluid. The MP release levels ranked from PGA 100 > > PGA 90 > PGA 75 > > PP ∼ PA. A typical PGA 100 suture released 0.63 ± 0.087 million micro (MPs > 1 µm) and 1.96 ± 0.04 million nano (NPs, 200-1000 nm) plastic particles per centimeter. In contrast, no MPs were released from the nonabsorbable sutures under the same conditions. PGA that was co-blended with 10-25% L-lactide or epsilon-caprolactone resulted in a two orders of magnitude lower level of MP release. These results underscore the need to assess the release of nano- and microplastics from medical polymers while applied in the human body and to evaluate possible risks to human health.

Preservation effects of photodynamic inactivation-mediated antibacterial film on storage quality of salmon fillets: Insights into protein quality.

The preservation effects of a photodynamic inactivation (PDI)-mediated polylactic acid/5-aminolevulinic acid (PLA/ALA) film on the storage quality of salmon fillets were investigated. Results showed that the PDI-mediated PLA/ALA film could continuously generate reactive oxygen species by consuming oxygen to inactivate native pathogens and spoilage bacteria on salmon fillets. Meanwhile, the film maintained the content of muscle proteins and their secondary and tertiary structures, as well as the integrity of myosin by keeping the activity of Ca2+-ATPase, all of which protected the muscle proteins from degradation. Furthermore, the film retained the activity of total superoxide dismutase (T-SOD), suppressed the accumulation of lipid peroxides (e.g., MDA), which greatly inhibited four main types of protein oxidations. As a result, the content of flavor amino acids and essential amino acids in salmon fillets was preserved. Therefore, the PDI-mediated antimicrobial packaging film greatly preserves the storage quality of aquatic products by preserving the protein quality.

Depth profiling of PLGA copolymer in a novel biomedical bilayer using confocal Raman spectroscopy.

Confocal Raman spectroscopy was undertaken to identify separate layers of PLGA and gentamicin sulfate (GS) coatings on a titanium alloy substrate for a novel drug-delivery system. Additionally, it was found that it was possible to measure the layer thickness and uniformity of the PLGA accurately by detecting intensity and wavelength changes in the vibrational bands of the copolymer bonds. Further analysis of the materials was done using FIB, SEM/EDX, and profilometry; these techniques were used to confirm the findings of the Raman data. It was determined that the substrate was extremely rough and therefore the coating was not uniform in thickness but the materials were uniformly dispersed. Most importantly, two distinct GS and PLGA layers were present.

Effects of microplastics on seed germination and seedling physiological characteristics of Spinacia oleracea under alkali stress.

Microplastics, a type of new environmental pollutant, have received much attention for their negative effects on organisms and environment. We examined the effects of microplastics on seed germination and seedling physiological characteristics of spinach (Spinacia oleracea) under alkali stress, taking polystyrene microspheres with a diameter of 100 nm (200, 400, 800, 1600 mg·L-1) as the microplastic treatment, and mixed NaHCO3 and Na2CO3 as alkaline salt solution (5, 10, 20, 40 mmol·L-1) according to the molar ratio of 1:1. The results showed that the presence of MPs (≥400 mg·L-1) inhibited seed germination, and that the length of roots and shoots increased at low while decreased at high concentration of MPs. Different concentrations of alkali alone could inhibit seed germination, root and bud elongation. With the increases of MPs concentration, SOD activity of spinach seedlings gradually decreased, while POD activity firstly increased and then decreased, and chlorophyll content increased at low concentration (200 mg·L-1) and decreased significantly at medium and high concentration (≥400 mg·L-1). Different alkali stresses reduced chlorophyll content of spinach seedlings, and the effects on SOD and POD were 'promotion at low concentration and inhibition at high'. In the treatments of microplastics (200, 800 mg·L-1) and alkali (5, 20 mmol·L-1) combined exposure, germination of spinach seeds was inhibited, and chlorophyll content decreased. The activities of SOD and POD in spinach seedlings were reduced under the combined exposure except the treatment of 200 mg·L-1 MPs and 5 mmol·L-1 alkali. Compared to the alkali stress, the combination of low concentration of MPs (200 mg·L-1) and alkali could improve germination rate, germination index, germination vigor and vigor index of seeds, and significantly promoted the elongation of roots and shoots, while the addition of high concentration of MPs (800 mg·L-1) reduced the germination rate, germination index, germination vigor and vigor index of seeds and inhibited the growth of roots and buds. The different concentrations of combined exposure inhibited the activities of SOD and POD and decreased the content of chlorophyll in spinach seedlings.

Covalent Grafting of 5-Aminolevulinic Acid onto Polylactic Acid Films and Their Photodynamic Potency in Preserving Salmon.

A novel photodynamic inactivation (PDI)-mediated antimicrobial film of polylactic acid/5-aminolevulinic acid (PLA/ALA) was successfully fabricated by a covalent grafting method using low-temperature plasma. The chemical structure, surface morphology, hydrophilic ability, and mechanical and barrier properties of the films were characterized, and their antibacterial, anti-biofilm potency and preservation effects on ready-to-eat salmon were investigated during storage. Results showed that the amino group of ALA was covalently grafted with the carboxyl group on the surface of PLA after the plasma treatment, with the highest grafting rate reaching ∼50%. The fabricated PLA/ALA films displayed an enhanced barrier ability against water vapor and oxygen. Under blue light-emitting diode illumination, the PLA/ALA films generated massive reactive oxygen species from the endogenous porphyrins in cells induced by ALA and then fatally destroyed the cell wall of planktonic cells and the architectural structures of sessile biofilms of the pathogens (Listeria monocytogenes and Vibrio parahaemolyticus) and spoilage bacterium (Shewanella putrefaciens). More importantly, the PDI-mediated PLA/ALA films potently inhibited 99.9% native bacteria on ready-to-eat salmon and significantly suppressed the changes of its drip loss, pH, and lipid oxidation (MDA) during storage, and on this basis, the shelf life of salmon was extended by 4 days compared with that of the commercial polyethylene film. Therefore, the PDI-mediated PLA/ALA films are valid in inactivating harmful bacterial and preserving the quality of seafood.

Polyethylene microplastics cause apoptosis via the MiR-132/CAPN axis and inflammation in carp ovarian.

Microplastics (MPs) are widely distributed pollutants in the environment and accumulate in the aquatic environment due to human activities. Carp, a common edible aquatic organism, has been found to accumulate MPs in body. MicroRNA (miRNAs) is a non-coding short RNA that regulates protein expression by binding to target genes in various physiological processes such as proliferation, differentiation and apoptosis. The ovary is a crucial role in carp reproduction. In this study, we established a model of carp exposed to polyethylene microplastics (PE-MPs) in the aquatic environment to investigate the specific mechanism of PE-MPs causing ovarian injury and the involvement of miR-132/calpain (CAPN) axis. H&E stained sections revealed that PE-PMs induced inflammation in ovarian tissues and impaired oocyte development. TUNEL analysis showed an increased rate of apoptosis in ovarian cells treated with PE-PMs. RT-PCR and Western Blot assays confirmed that exposure to PE-MPs significantly decreased miR-132 expression while increasing CAPN expression at both mRNA and protein levels. The concentration of calcium ions was significantly increased in tissues, leading to CAPN enzyme activity increase. The expression of mitochondrial damage-related genes (bax, AIF, cyt-c, caspase-7, caspase-9, and caspase-3) was higher while the expression of anti-apoptotic genes (bcl-2 and bcl-xl) was lower. Protein levels of bax, AIF, caspase-3, bcl-2 and bcl-xl changed accordingly with the genetic alterations. Additionally, we discovered that PE-MPs can activate the p65 factor through the TRAF6/NF-kB pathway resulting in elevated production of pro-inflammatory factors IL-6, IL-1ß and TNF-a which contribute to ovarian inflammation development. This study investigates the impact of PE-MPs on carp ovarian function and provides insights into miRNAs' role and their target genes.

[Spatial variability of soil available potassium in rubber plantation based on coKriging]. / åŸºäºŽååŒå ‹é‡Œæ ¼çš„æ©¡èƒ¶å›­åœŸå£¤é€Ÿæ•ˆé’¾ç©ºé—´å˜å¼‚æ€§.

The coKriging method of selecting effective variables is helpful to improve the spatial prediction accuracy of soil available potassium in county-scale rubber plantation, which is of significance in precision fertilization management of rubber plantation. In this study, we analyzed the spatial variability characteristics of soil available potassium in 0-20 cm layer in the rubber plantation of Baisha County, Hainan Province, by geostatistics. The significant characteristic variables were screened by correlation analysis, and the spatial interpolation precisions of coKriging (COK) of different variables were compared. The results showed that the average soil available potassium content in the study area was 44.65 g·kg-1, generally at a state of shortage. The variable coefficient was 52.6%, which was a moderate intensity of variation. The nugget effect was 12.5%, with a strong spatial autocorrelation. The organic matter and elevation were closely related to soil available potassium content. The COK spatial interpolation prediction precisions of the three covariates of organic matter (COK1), elevation (COK2), and organic matter+elevation (COK3) were all higher than ordinary Kriging (OK), and the fitting precision of the cross-validation model was COK1>COK3>COK2>OK. The fitting precision was not proportional to the number of covariates selected. Selecting more correlated covariates was more conducive to reflecting the spatial heterogeneity of soil properties. The soil available potassium content was higher in the northwest and lower in the central and eastern regions, which provided a theoretical basis for the further development of soil potassium management in rubber plantations.

A review of potential human health impacts of micro- and nanoplastics exposure.

This systematic review aims to summarize the current knowledge on biological effects of micro- and nanoplastics (MNPs) on human health based on mammalian systems. An extensive search of the literature led to a total of 133 primary research articles on the health relevance of MNPs. Our findings revealed that although the study of MNP cytotoxicity and inflammatory response represents a major research theme, most studies (105 articles) focused on the effects of polystyrene MNPs due to their wide availability as a well characterised research material that can be manufactured with a large range of particle sizes, fluorescence labelling as well as various surface modifications. Among the 133 studies covered in this review, 117 articles reported adverse health effects after being exposed to MNPs. Mammalian in vitro studies identified multiple biological effects including cytotoxicity, oxidative stress, inflammatory response, genotoxicity, embryotoxicity, hepatotoxicity, neurotoxicity, renal toxicity and even carcinogenicity, while rodent in vivo models confirmed the bioaccumulation of MNPs in the liver, spleen, kidney, brain, lung and gut, presenting adverse effects at different levels including reproductive toxic effects and trans-generational toxicity. In contrast, the remaining 16 studies indicated an insignificant impact of MNPs on humans. A few studies attempted to investigate the mechanisms or factors driving the toxicity of MNPs and identified several determining factors including size, concentration, shape, surface charge, attached pollutants and weathering process, which, however, were not benchmarked or considered by most studies. This review demonstrates that there are still many inconsistencies in the evaluation of the potential health effects of MNPs due to the lack of comparability between studies. Current limitations hindering the attainment of reproducible conclusions as well as recommendations for future research directions are also presented.

[Effects of manure substituting chemical nitrogen fertilizer on rubber seedling growth and soil environment.] / 有机肥替代化学氮肥对橡胶幼苗生长和土壤环境的影响.

The substitution of manure for chemical nitrogen fertilizers has great impacts on the growth of rubber seedlings and soil environment, with implications for rubber cultivation and transplantation and soil environment improvement. In this study, rubber seedlings of thermal research '7-33-97' strain were cultivated under four treatments: No fertilizer application (CK), only application of chemical fertilizer (N), manure replacing 50% chemical fertilizer (M+N), and manure replacing 100% chemical fertilizer (M). Plants parameters (including plant height, basal diameter, biomass, and chlorophyll), soil physicochemical properties (including pH, soil organic carbon and nitrogen, soil enzyme activities), and their relationships were investigated. The results showed that plant height, basal diameter, biomass, and chlorophyll content in the M+N and M treatments were significantly higher, while underground biomass and root-shoot ratio were significantly lower than those of in N treatment. Compared with CK, soil pH was significantly increased in the M treatment, decreased in the N treatment, and was not changed in the M+N treatment. Soil ammonium and nitrate content in the M+N and M treatments were significantly lower, while soil organic carbon content, the activity of ß-1,4-glucosidase (BG), ß-1,4-N-acetylglucosaminidase (NAG) and leucine aminopeptidase (LAP) were significantly higher than those of in N treatment. Results of correlation analysis showed that soil pH was negatively correlated with soil ammonium and nitrate content, but positively correlated with BG and NAG activities. The structural equation model analysis showed that soil pH had significant positive effects on seedling quality index, while nitrate content had significant negative effects, and soil enzyme activities had no significant effect. Those results indicated that soil pH and nitrate content were the important driving factors on the growth of rubber seedlings. The manure replacing of 50% and 100% chemical nitrogen fertilizer could promote rubber seedlings growth, improve soil environment, and promote sustainable development of rubber production in Danzhou City, Hainan Province.

The influence of drinking water constituents on the level of microplastic release from plastic kettles.

Microplastic (MP) release from household plastic products has become a global concern due to the high recorded levels of microplastic and the direct risk of human exposure. However, the most widely used MP measurement protocol, which involves the use of deionized (DI) water, fails to account for the ions and particles present in real drinking water. In this paper, the influence of typical ions (Ca2+/HCO3-, Fe3+, Cu2+) and particles (Fe2O3 particles) on MP release was systematically investigated by conducting a 100-day study using plastic kettles. Surprisingly, after 40 days, all ions resulted in a greater than 89.0% reduction in MP release while Fe2O3 particles showed no significant effect compared to the DI water control. The MP reduction efficiency ranking is Fe3+ ≈ Cu2+ > Ca2+/HCO3- > > Fe2O3 particles ≈ DI water. Physical and chemical characterization using SEM-EDX, AFM, XPS and Raman spectroscopy confirmed Ca2+/HCO3-, Cu2+ and Fe3+ ions are transformed into passivating films of CaCO3, CuO, and Fe2O3, respectively, which are barriers to MP release. In contrast, there was no film formed when the plastic was exposed to Fe2O3 particles. Studies also confirmed that films with different chemical compositions form naturally in kettles during real life due to the different ions present in local regional water supplies. All films identified in this study can substantially reduce the levels of MP release while withstanding the repeated adverse conditions associated with daily use. This study underscores the potential for regional variations in human MP exposure due to the substantial impact water constituents have on the formation of passivating film formation and the subsequent release of MPs.

Photoinduced graft polymerization of 2-methacryloyloxyethyl phosphorylcholine on silicone hydrogels for reducing protein adsorption.

The biomimetic synthetic methacrylate monomer containing a phosphorylcholine group, 2-methacryloyloxyethyl phosphorylcholine (MPC), has been widely used to improve the surface property of biomaterials. In the current report, both hydrophilic and antifouling surfaces were prepared on silicone hydrogels with MPC grafted by UV-induced free radical polymerization. The MPC-grafted silicone hydrogels were characterized by graft yield and static water contact angle (SCA) measurements. According to the results, the graft yield reached a maximum at 5 min of UV exposure time and 8 wt% MPC concentration. The modified silicone hydrogels possessed hydrophilic surfaces with the lowest water contact angle of 20º. The oxygen permeability of the MPC-grafted silicone hydrogels was as high as the unmodified silicone hydrogel. The mechanical property of silicone hydrogels was maintained at about 95% of the tensile strength and elastic modulus after the MPC grafting. The results of the in vitro single protein adsorption on the MPC-grafted silicone hydrogels were in agreement with the SCA measurements. The smaller the water contact angle, the greater was the protein repelling ability. The MPC-grafted silicone hydrogel is expected to be a novel biomaterial which possesses excellent surface hydrophilicity, antifouling property, oxygen permeability and mechanical property.

Sampling, Identification and Characterization of Microplastics Release from Polypropylene Baby Feeding Bottle during Daily Use.

Microplastics (MPs) are becoming a global concern due to the potential risk to human health. Case studies of plastic products (i.e., plastic single-use cups and kettles) indicate that MP release during daily use can be extremely high. Precisely determining the MP release level is a crucial step to identify and quantify the exposure source and assess/control the corresponding risks stemming from this exposure. Though protocols for measuring MP levels in marine or freshwater has been well developed, the conditions experienced by household plastic products can vary widely. Many plastic products are exposed to frequent high temperatures (up to 100 °C) and are cooled back to room temperature during daily use. It is therefore crucial to develop a sampling protocol that mimics the actual daily-use scenario for each particular product. This study focused on widely used polypropylene-based baby feeding bottles to develop a cost-effective protocol for MP release studies of many plastic products. The protocol developed here enables: 1) prevention of the potential contamination during sampling and detection; 2) realistic implementation of daily-use scenarios and accurate collection of the MPs released from baby feeding bottles based on WHO guidelines; and 3) cost-effective chemical determination and physical topography mapping of MPs released from baby feeding bottles. Based on this protocol, the recovery percentage using standard polystyrene MP (diameter of 2 µm) was 92.4-101.2% while the detected size was around 102.2% of the designed size. The protocol detailed here provides a reliable and cost-effective method for MP sample preparation and detection, which can substantially benefit future studies of MP release from plastic products.

Sodium fluoride exposure triggered the formation of neutrophil extracellular traps.

In recent years, numerous studies paid more attention to the molecular mechanisms associated with fluoride toxicity. However, the detailed mechanisms of fluoride immunotoxicity in bovine neutrophils remain unclear. Neutrophil extracellular traps (NETs) is a novel immune mechanism of neutrophils. We hypothesized that sodium fluoride (NaF) can trigger NETs activation and release, and investigate the related molecular mechanisms during the process. We exposed peripheral blood neutrophils to 1 mM NaF for 120 min in bovine neutrophils. The results showed that NaF exposure triggered NET-like structures decorated with histones and granule proteins. Quantitative measurement of NETs content correlated positively with the concentration of NaF. Mechanistically, NaF exposure increased reactive oxygen species (ROS) levels and phosphorylation levels of ERK, p38, whereas inhibiting the activities of superoxide dismutase (SOD) and catalase (CAT) compared with control neutrophils. NETs formation is induced by NaF and this effect was inhibited by the inhibitors diphenyleneiodonium chloride (DPI), U0126 and SB202190. Our findings described the potential importance of NaF-triggered NETs related molecules, which might help to extend the current understanding of NaF immunotoxicity.

VEGF-PLGA controlled-release microspheres enhanced angiogenesis in encephalomyosynangiosis-based chronic cerebral hypoperfusion.

Treatments enhancing angiogenesis for chronic cerebral hypoperfusion (CCH) are still in the research stage. Although encephalomyosynangiosis (EMS) is a common indirect anastomosis for the treatment of CCH, the effectiveness to promote angiogenesis is not satisfactory. Vascular endothelial growth factors (VEGF) is a cytokine found to specifically act directly on vascular endothelial cells, promote neovascularization, and enhance capillary permeability. However, the short half life and unstable property of VEGF underlies the need to explore available delivery system. In this study, poly (lactide-co-glycolide) (PLGA) was used to prepare VEGF controlled-release microspheres. In vitro and in vivo analysis of release kinetics showed that the microspheres could release VEGF continuously within 30 days. Then, modified chronic cerebral hypoperfusion rat model was established by ligation of bilateral internal carotid artery and one vertebral artery. At 14 days after ischemia, the EMS and the VEGF microspheres injection were performed. At 30 days after the injection, the result of Morris water maze displayed that combinating VEGF microspheres and EMS significantly ameliorated cognitive deficit after ischemia. We observed that combinating VEGF microspheres and EMS could further significantly increase cerebral blood flow. We speculated that this enhancement of cerebral blood flow was attributed to more angiogenesis induced by combination of VEGF microspheres and EMS, which verified by more collateral circulation with cerebral angiography and higher expression of CD31 or α-SMA. Our study demonstrated that combinating VEGF-PLGA controlled-release microspheres could significantly promote angiogenesis in EMS-based CCH rats model, providing new ideas for clinical treatment of CCH.

Bamboo (Phyllostachys pubescens) as a Natural Support for Neutral Protease Immobilization.

Lignin polymers in bamboo (Phyllostachys pubescens) were decomposed into polyphenols at high temperatures and oxidized for the introduction of quinone groups from peroxidase extracted from bamboo shoots and catalysis of UV. According to the results of FT-IR spectra analysis, neutral proteases (NPs) can be immobilized on the oxidized lignin by covalent bonding formed by amine group and quinone group. The optimum condition for the immobilization of NPs on the bamboo bar was obtained at pH 7.0, 40 °C, and duration of 4 h; the amount of immobilized enzyme was up to 5 mg g-1 bamboo bar. The optimal pH for both free NP (FNP) and INP was approximately 7.0, and the maximum activity of INP was determined at 60 °C, whereas FNP presented maximum activity at 50 °C. The Km values of INP and FNP were determined as 0.773 and 0.843 mg ml-1, respectively; INP showed a lower Km value and Vmax, than FNP, which demonstrated that INP presented higher affinity to substrate. Compared to FNP, INP showed broader thermal and storage stability under the same trial condition. With respect to cost, INP presented considerable recycling efficiency for up to six consecutive cycles.

Role of extracellular polymeric substances in the acute inhibition of activated sludge by polystyrene nanoparticles.

Microplastics and nanoplastics in aquatic systems have become a global concern because of their persistence and adverse consequences to ecosystems and potentially human health. Though wastewater treatment plants (WWTPs) are considered a potential source of microplastics in the environment, the role of extracellular polymeric substances (EPS) of activated sludge on the fate of nanoplastics is not clear. In this study, the role of EPS in the influence of polystyrene nanoparticles (PS-NPs) on the endogenous respiration of activated sludge was investigated for the first time. The results showed that the acute inhibition of activated sludge by PS-NPs was enhanced with increasing PS-NPs concentration. X-ray photoelectron spectroscopy (XPS) results indicate that the functional groups involved in the interactions between PS-NPs and EPS were carbonyl and amide groups and the side chains of lipids or amino acids. Furthermore, the Fourier transform infrared (FTIR) spectroscopy results show that the protein secondary structures in EPS were changed by PS-NPs and lead to the bioflocculation of activated sludge, which provides a better understanding on the fate of nanoplastics in WWTPs.