Production of surfactant-stable keratinase from Bacillus cereus YQ15 and its application as detergent additive.

BACKGROUND: With the growing concern for the environment, there are trends that bio-utilization of keratinous waste by keratinases could ease the heavy burden of keratinous waste from the poultry processing and leather industry. Especially surfactant-stable keratinases are beneficial for the detergent industry. Therefore, the production of keratinase by Bacillus cereus YQ15 was improved; the characterization and use of keratinase in detergent were also studied. RESULTS: A novel alkaline keratinase-producing bacterium YQ15 was isolated from feather keratin-rich soil and was identified as Bacillus cereus. Based on the improvement of medium components and culture conditions, the maximum keratinase activity (925 U/mL) was obtained after 36 h of cultivation under conditions of 35 °C and 160 rpm. Moreover, it was observed that the optimal reacting temperature and pH of the keratinase are 60 °C and 10.0, respectively; the activity was severely inhibited by PMSF and EDTA. On the contrary, the keratinase showed remarkable stability in the existence of the various surfactants, including SDS, Tween 20, Tween 60, Tween 80, and Triton X-100. Especially, 5% of Tween 20 and Tween 60 increased the activity by 100% and 60%, respectively. Furtherly, the keratinase revealed high efficiency in removing blood stains. CONCLUSION: The excellent compatibility with commercial detergents and the high washing efficiency of removing blood stains suggested its suitability for potential application as a bio-detergent additive.

[Effects of end time of night feeding on the growth and development of infants/toddlers].

OBJECTIVE: To investigate the effects of end time of night feeding on body height, body weight, nutritional status, and prevalence rate of dental caries in children at the age of 30 months. METHODS: A total of 416 children who were born from January 2014 to September 2015 and had completed a physical examination as required were enrolled. During the physical examination performed at the age of 30 months, the comprehensive child care record and a self-made questionnaire were used. The children who continued to receive night feeding after the age of 6 months were enrolled as study group (n=269), and those for whom night feeding was ended at the age of 6 months were enrolled as control group (n=147). The two groups were compared in terms of body height, body weight, incidence rate of overweight/obesity, and prevalence rate of dental caries at the age of 30 months. RESULTS: Compared with the control group, the study group had a significantly lower body height (92.4±3.0 cm vs 93.3±2.8 cm; P<0.05), a significantly higher incidence rate of overweight/obesity (23.8% vs 12.2%; P<0.05), and a significantly higher prevalence rate of dental caries (14.9% vs 7.5%; P<0.05) at the age of 30 months. CONCLUSIONS: Night feeding continued after the age of 6 months can affect the growth and development of infants/toddlers, cause overnutrition, and increase the prevalence rate of dental caries.

Anion Channel Inhibitor NPPB-Inhibited Fluoride Accumulation in Tea Plant (Camellia sinensis) Is Related to the Regulation of Ca²âº, CaM and Depolarization of Plasma Membrane Potential.

Tea plant is known to be a hyper-accumulator of fluoride (F). Over-intake of F has been shown to have adverse effects on human health, e.g., dental fluorosis. Thus, understanding the mechanisms fluoride accumulation and developing potential approaches to decrease F uptake in tea plants might be beneficial for human health. In the present study, we found that pretreatment with the anion channel inhibitor NPPB reduced F accumulation in tea plants. Simultaneously, we observed that NPPB triggered Ca(2+) efflux from mature zone of tea root and significantly increased relative CaM in tea roots. Besides, pretreatment with the Ca(2+) chelator (EGTA) and CaM antagonists (CPZ and TFP) suppressed NPPB-elevated cytosolic Ca(2+) fluorescence intensity and CaM concentration in tea roots, respectively. Interestingly, NPPB-inhibited F accumulation was found to be significantly alleviated in tea plants pretreated with either Ca(2+) chelator (EGTA) or CaM antagonists (CPZ and TFP). In addition, NPPB significantly depolarized membrane potential transiently and we argue that the net Ca(2+) and H⁺ efflux across the plasma membrane contributed to the restoration of membrane potential. Overall, our results suggest that regulation of Ca(2+)-CaM and plasma membrane potential depolarization are involved in NPPB-inhibited F accumulation in tea plants.

Synergism of jet milling and deep eutectic solvent pretreatment on grapevine lignin fractionation and enhancing enzymatic hydrolysis.

Herein, we investigated the synergistic effects of jet milling (JM) and deep eutectic solvent (DES) pretreatment on the fractionation of grapevine lignin and the consequent enhancement of enzymatic hydrolysis. Grapevine, a substantial byproduct of the wine industry, was subjected to JM pretreatment to produce finely powdered particles (median diameter D50 = 98.90), which were then further treated with acidic ChCl-LA and alkaline K2CO3-EG DESs. The results revealed that the combined JM + ChCl-LA pretreatment significantly increased the cellulose preservation under optimal conditions (110 °C, 4 h, and 20 % water content), achieving removal rates of 74.18 % xylan and 66.05 % lignin, respectively. The pretreatment temperature and inhibitor production were reduced, resulting in a remarkable threefold increase in glucose yield compared to untreated samples. Moreover, the structural analysis of the pretreated lignin indicated an enrichment of phenolic units, leading to enhanced antioxidant and antibacterial activities, particularly in the JM pretreated samples. These findings underscore the promising potential of the synergistic JM and DES pretreatment in facilitating the efficient utilization of grapevine lignocellulosic biomass for sustainable biorefinery technologies.

Molecular and morphological evidence supports the resurrection of Chrysospleniumguangxiense H.G.Ye & Gui C.Zhang (Saxifragaceae).

Chrysospleniumguangxiense H.G.Ye & Gui C.Zhang was first described as a new species in 1994 but later synonymized in the Flora of China treatment with C.glossophyllum H.Hara. Plastid genomes and nrDNA sequences were used to infer the phylogenetic relationships of selected taxa in Chrysosplenium. Our phylogenetic analyses revealed that C.guangxiense belongs to sect. Alternifolia, is closely related to Chrysospleniumhydrocotylifolium H.Lév. & Vaniot but distant from C.glossophyllum. Morphologically, C.guangxiense could be easily distinguished from C.glossophyllum by having robust rhizomes, basal leaves with a long cuneate base and fewer teeth in the margin, curled sepal margins, and red, larger seeds. It could also be easily distinguished from C.hydrocotylifolium by possessing long elliptic leaves and a long cuneate leaf base. Along with the phylogenetic studies, the complete plastid genome of C.guangxiense was also reported. The plastid genome was 154,004 bp in length and comprised two inverted repeats (IRs) of 28,120 bp, separated by a large single-copy of 80,646 bp and a small single-copy of 17,118 bp. A total of 111 functional genes were discovered, comprising 78 protein-coding genes, 29 tRNA genes, and four rRNA genes. Based on assessment of morphological and molecular data Chrysospleniumguangxiense H.G.Ye & Gui C.Zhang is resurrected from C.glossophyllum H.Hara at species level. A global conservation assessment classifies C.guangxiense as Vulnerable (VU).

Acidic and alkaline deep eutectic solvents (DESs) pretreatment of grapevine: Component analysis, characterization, lignin structural analysis, and antioxidant properties.

Deep eutectic solvents (DESs) have been extensively applied to pretreat lignocellulose; however, comparative research on acidic and alkaline DES pretreatment is relatively lacking. Herein, pretreatment of grapevine agricultural by-products with seven DESs were compared in terms of removal of lignin and hemicellulose and component analysis of the pretreated residues. Among the tested DESs, both acidic choline chloride-lactic (CHCl-LA), and alkaline potassium carbonate-ethylene glycol (K2CO3-EG) were effective in delignification. Thereafter, the CHCl-LA and K2CO3-EG extracted lignin was compared by analyzing their physicochemical structure changes and antioxidant properties. The results showed that the thermal stability, molecular weight, and phenol hydroxyl percentage of CHCl-LA lignin were inferior to K2CO3-EG lignin. It was found that the high antioxidant activity of K2CO3-EG lignin was mainly attributed to the abundant phenol hydroxyl, guaiacyl (G), and para-hydroxy-phenyl (H). By comparing acidic and alkaline DES pretreatments and their lignin nuances in biorefining, novel insights are derived for the scheduling and selection of DES for lignocellulosic pretreatment.

Preparation of polyvinyl alcohol/chitosan nanofibrous films incorporating graphene oxide and lanthanum chloride by electrospinning method for potential photothermal and chemical synergistic antibacterial applications in wound dressings.

Electrospun fibres have been widely used as skin dressings due to their unique structur. However, due to the lack of intrinsic antimicrobial activity, it is easy for the wound to become infected. Bacterial infection, which leads to chronic inflammation, severely hinders the normal process of skin regeneration. In this study, a polyvinyl alcohol/chitosan (PVA/CS) composite films with chemical sterilization and near-infrared (NIR) photothermal antibacterial activity was fabricated by electrospinning. Graphene oxide (GO), a photosensitiser, was incorporated into the films, and lanthanum chloride (Lacl3) as a chemical antibacterial agent was also doped in the electrospun films. The structure, morphology, mechanical properties, wettability, and antimicrobial and photothermal antibacterial activity of the PVA/CS-based fibre films were investigated. The results showed that the addition of Lacl3 to the PVA/CS/GO nanofibres (PVA/CS/GO-La) improved the hydrophilicity, tensile strength and resistance to elastic deformation of the nanofibres. The PVA/CS/GO-La12.5 mM sample exhibited the best antibacterial performance, showing high inhibition against Staphylococcus aureus (82% antibacterial efficacy) and Escherichia coli (99.7% antibacterial efficacy). Furthermore, the antibacterial efficacy of the films surface was further enhanced after exposure to NIR light (808 nm, 0.01 W) for 20 min. In addition, the nanofibre films showed no cytotoxicity against human skin fibroblasts (HSFs), indicating its potential application in the field of broad-spectrum antibacterial materials.

[Surface enhanced Raman spectroscopy (SERS) of saliva for the diagnosis of lung cancer].

Surface enhanced Raman spectroscopy (SERS) has shown the advantage of detecting low concentration biofluids presently. Saliva SERS of 21 lung cancer patients and 22 normal people were measured and differentiated in the present paper. Intensities of most peaks of lung cancer patients are weaker than that of normal people, while some stronger but with a small change rate. Those peaks were assigned to proteins and nucleic acids which indicate a corresponding decrease of those substances in saliva. Principal component analysis (PCA) and linear discriminant analysis (LDA) were used to deduce and discriminate the two groups of data, resulting in accuracy, sensitivity, and specificity being 84%, 94%, and 81%, respectively.

Auto-fluorescence of cellulose paper with spatial solid phrase dispersion-induced fluorescence enhancement behavior for three heavy metal ions detection.

Auto-fluorescence of cellulose paper is often considered as an interfering fluorescence, which directly impedes the cellulose paper as a substrate material. This paper creatively explored the composition and properties of auto-fluorescence, and lignosulfonate was primarily speculated as the main source of auto-fluorescence. Surprisingly, its spatial solid phrase dispersion-induced fluorescence enhancement behavior was found. Then, cellulose paper was modified with Mn-doped ZnS quantum dots, and the prepared ratiometric fluorescent paper chip has good performances on morphology, stability, and fluorescence properties. Besides, the paper chip exhibited different fluorescence responses to three heavy metal ions in water sample. The limit of detection for Cd2+, Hg2+ and Pb2+ reached 1.61 nM, 0.01 nM, and 0.02 nM, respectively. In short, the molecular simulation results theoretically proved that heavy metal ions owned substitution affinity with lignosulfonate. Ultimately, this study was the first attempt to utilize paper-based auto-fluorescence, which could better accelerate the development of paper-based chips.

Disrupted microbiota-barrier-immune interaction in phthalates-mediated barrier defect in the duodenum.

As one of the most used phthalates, Di (2-ethylhexyl) phthalate (DEHP) is a widespread environmental contaminant. Extremely persistent plastic can enter the food chain of animals through the aquatic environment, affect metabolic pathways and cause damage to the digestive system. But the molecular mechanism of its toxic effects on the duodenum in birds has not been elucidated. To investigate the toxicity of phthalates in the duodenum, quails were gavaged with 250, 500, and 750 mg/kg doses of DEHP for 45 days, and water and oil control groups were retained. This study revealed that subchronic exposure to DEHP could lead to duodenal barrier defect in quail. The damage to duodenum was reflected in a reduction in V/C and tight junction proteins. Moreover, DEHP also led to a breakdown of antimicrobial defenses through the flora derangement, which acted as a biological barrier. The massive presence of Lipopolysaccharide (LPS) led to the activation of TLR4 receptors. In addition, DEHP activated oxidative stress, which synergized the inflammatory response induced by the TLR4-NFκB pathway, and further promoted duodenum damage. This study provides a base for the further effect of phthalates on the microbiota-barrier-immune interaction.

The toxic effects of polystyrene microplastics on freshwater algae Chlorella pyrenoidosa depends on the different size of polystyrene microplastics.

Microplastics (MPs) are persistent environmental contaminants. The toxic effects of MPs on aquatic organisms have raised increasing concerns, but their toxic effects on aquatic phytoplankton has not been thoroughly investigated. In the present study, the toxic effects of two sizes MPs (1 µm and 5 µm) on Chlorella pyrenoidosa at 2, 10, 50 mg/L were explored for 1, 5, 10 days. The growth ratio, photosynthetic pigments content, extracellular polymeric substances content, soluble protein content, MDA content and relative expression of genes related to photosynthesis and energy metabolism were measured. These results indicated that 1 µm MP could significantly inhibit the growth of C. pyrenoidosa. Compared with the control group, 1 µm MP significantly reduced the photosynthetic pigment content, induced oxidative stress and disrupted the cell membrane integrity of C. pyrenoidosa. At the molecular level, 1 µm MP altered the transcript levels of genes related to photosynthesis and energy metabolism. Scanning electron microscopy and fluorescent images showed that MPs aggregation with C. pyrenoidosa may be the main reason for the toxic effects of MPs. These results will provide new insight into the toxicity of different MPs on aquatic phytoplankton, and evaluate the risks caused by MPs in aquatic environments.

Responsive Hydrogels Based on Triggered Click Reactions for Liver Cancer.

Globally, liver cancer, which is one of the major cancers worldwide, has attracted the growing attention of technological researchers for its high mortality and limited treatment options. Hydrogels are soft 3D network materials containing a large number of hydrophilic monomers. By adding moieties such as nitrobenzyl groups to the network structure of a cross-linked nanocomposite hydrogel, the click reaction improves drug-release efficiency in vivo, which improves the survival rate and prolongs the survival time of liver cancer patients. The application of a nanocomposite hydrogel drug delivery system can not only enrich the drug concentration at the tumor site for a long time but also effectively prevents the distant metastasis of residual tumor cells. At present, a large number of researches have been working toward the construction of responsive nanocomposite hydrogel drug delivery systems, but there are few comprehensive articles to systematically summarize these discoveries. Here, this systematic review summarizes the synthesis methods and related applications of nanocomposite responsive hydrogels with actions to external or internal physiological stimuli. With different physical or chemical stimuli, the structural unit rearrangement and the controlled release of drugs can be used for responsive drug delivery in different states.

Development and evaluation of a novel nano-scale vector for siRNA.

To synthesize a lipid-cationic polymer (LCP) containing brassidic acid side chain and to investigate its transfection efficiency and characteristics as a siRNA gene vector. The LCP was chemically synthesized and its nucleic acid binding capacity was determined by gel electrophoresis. HeLa-EGFP and TH1080-EGFP cell lines were transfected with siRNA against enhanced green fluorescent protein (EGFP) gene using a LCP to investigate the transfection efficiency. An MTT assay was performed to evaluate the cellular toxicity of the LCP vector. Its degradability and stability under acidic conditions were also investigated. The LCP vector possessed high DNA binding capacity. More than 73% of the cellular fluorescence was inhibited by the LCP-mediated transfection of siRNA against EGFP gene, indicating that vector had high transfection efficiency. Cellular viability was about 95% at the optimum transfection efficiency of LCP, suggesting that the cellular toxicity of LCP was very low. The LCP was also observed to be degradable; moreover, it could be easily stored at normal temperature. A gene vector used for the transfection of siRNA was successfully fabricated from synthesized LCP. Its numerous excellent properties entitle values for further scientific research.

[Initial immunoreaction study on PVA/n-HA + PA66 biological composite material after being implanted into animal body].

OBJECTIVE: To observe immunoreaction to PVA/n-HA+ PA66 biological composite material after being implanted into animal body. METHODS: PVA/n-HA+PA66 composite materials were implanted into mouse subcutaneous tissue. Histologial examination was performed at 2, 4, 6 weeks, the amount of CD3, CD4+, CD8+ in blood and IL-1, IL-6, TNF-alpha in spleen cells was measured at the same time. RESULTS: The amount of CD3, CD4+, CD8+ in mouse blood and IL-1, IL-6, TNF-alpha in mouse spleen cells were not different with the control groups at 2, 4, 6 weeks (P > 0.05). The fibrous tissue and some blood vessels were found growing into the porous materials, which resulted in composite materials intergration of proliferative structure. The reject reaction was not found. CONCLUSION: PVA/n-HA+PA66 biological composite material does not cause immune rejection after being implanted into animal body, verifying its good biocompatibility.

[Porous polyvinyl alcohol hydrogel composite prepared and studied initially for biocompatibility].

OBJECTIVE: To investigate the feasibility of porous polyvinyl alcohol hydrogel (PVA-H) composite, for tissue engineering scaffold. METHODS: The approach of emulsifier-foaming, freeze-drying and surfactant-cleaning method was developed to gain three kinds of porous scaffolds: porous polyvinyl alcohol (PVA), porous polyvinyl alcohol/nano-hydroxyapatite (PVA/n-HA) and porous polyvinyl alcohol/chitosan (PVA/Cs). We checked the physical features of the above three kinds of materials and further investigated their biocompatibility by SEM image analysis, mechanics test, MTT and muscle implanting. RESULTS: Three kinds of materials had the similar character of high porosity (80%), but the PVA/n-HA had the smallest mean pore size (154.5 microm) and the PVA had the biggest tensile strength (0.60 Mpa). MTT assay demonstrated three kinds of materials to have no toxicity, furthermore, PVA had better absorbance, compared with cell control group (P < 0.05). After muscle implantation, there were many muscular tissues growing into pores of PVA/Cs at the 4th week, although PVA/Cs had stronger inflammatory reaction. Other two kinds of materials had very small inflammatory reaction and there were many fibrous tissues growing into pores at the 1st, 4th, 12th week. CONCLUSION: The features of materials are changed after adding n-HA or Cs, moreover, porous PVA/Cs seems to have the activity of inducing muscle growth. Porous PVA and its composites may be applied to tissue engineering as a long-term or permanent scaffold due to their good biocompatibility, elasticity and hydrophilicity.

Nonwoven Polymer Nanofiber Coatings That Inhibit Quorum Sensing in Staphylococcus aureus: Toward New Nonbactericidal Approaches to Infection Control.

We report the fabrication and biological evaluation of nonwoven polymer nanofiber coatings that inhibit quorum sensing (QS) and virulence in the human pathogen Staphylococcus aureus. Our results demonstrate that macrocyclic peptide 1, a potent and synthetic nonbactericidal quorum sensing inhibitor (QSI) in S. aureus, can be loaded into degradable polymer nanofibers by electrospinning and that this approach can deposit QSI-loaded nanofiber coatings onto model nonwoven mesh substrates. The QSI was released over ∼3 weeks when these materials were incubated in physiological buffer, retained its biological activity, and strongly inhibited agr-based QS in a GFP reporter strain of S. aureus for at least 14 days without promoting cell death. These materials also inhibited production of hemolysins, a QS-controlled virulence phenotype, and reduced the lysis of erythrocytes when placed in contact with wild-type S. aureus growing on surfaces. This approach is modular and can be used with many different polymers, active agents, and processing parameters to fabricate nanofiber coatings on surfaces important in healthcare contexts. S. aureus is one of the most common causative agents of bacterial infections in humans, and strains of this pathogen have developed significant resistance to conventional antibiotics. The QSI-based strategies reported here thus provide springboards for the development of new anti-infective materials and novel treatment strategies that target virulence as opposed to growth in S. aureus. This approach also provides porous scaffolds for cell culture that could prove useful in future studies on the influence of QS modulation on the development and structure of bacterial communities.

Label-free immunosensor based on hyperbranched polyester for specific detection of α-fetoprotein.

A novel label-free immunosensor based on hyperbranched polyester nanoparticles with nitrite groups (HBPE-NO2), which were synthesized through a simple one-step chemical reaction, was first developed for specific detection of α-fetoprotein (AFP), the tumor marker for liver cancer. The obtained HBPE-NO2 nanoparticles (NPs) were characterized by the proton nuclear magnetic resonance spectroscopy (1H NMR), X-ray photoelectron spectroscopy (XPS) and X-ray diffraction (XRD). And the fabricated process of immunosensor was investigated by attenuated total reflection Fourier-transform infrared spectra (ATR-FTIR), static water contact angles, scanning electron microscope (SEM), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The electrochemical performances of the AFP immunosensor were studied. Results indicated the prepared HBPE-NO2-modified immunosensor showed excellent electrochemical properties and satisfactory accuracy for the detection of AFP of the real clinical samples that attributed to the properties of the HBPE-NO2 NPs, which had nanosized structure to increase the specific surface area and unique chemical reactivity for loading capacity of protein molecules. Construction of biosensors using the structure and properties of hyperbranched molecules will offer ideal electrode substrates, which provided more possibilities for the design of biosensor.

[Current treatment situation and progress on bone defect of collapsed tibial plateau fractures].

Characteristics of collapsed tibial plateau fracture determines that the joint surface must remain anatomical reduction,line of force in tibial must exist and internal fixation must be strong. However, while renewing articular surface smoothness, surgeons have a lot of problems in dealing with bone defect under the joint surface. Current materials used for bone defect treatment include three categories: autologous bone, allograft bone and bone substitutes. Some scholars think that autologous bone grafts have a number of drawbacks, such as increasing trauma, prolonged operation time, the limited source, bone area bleeding,continuous pain, local infection and anesthesia,but most scholars believe that the autologous cancellous bone graft is still the golden standard. Allograft bone has the ability of bone conduction, but the existence of immune responses, the possibility of a virus infection, and the limited source of the allograft cannot meet the clinical demands. Likewise, bone substitutes have the problem that osteogenesis does not match with degradation in rates. Clinical doctors can meet the demand of the patient's bone graft according to patient's own situation and economic conditions.

[In vitro and in vivo study of two kinds of long-circulating solid lipid nanoparticles containing paclitaxel].

AIM: To prepare long-circulating solid lipid nanoparticles containing paclitaxel with stearic acid, and investigate the in vitro and in vivo characterization of nanoparticles. METHODS: The method of "emulsion evaporation-solidification at low temperature" was used to prepare the stearic acid solid lipid nanoparticles containing paclitaxel. Its morphology was examined by transmission electron microscope. The HPLC method for determination of paclitaxel in nanoparticles or serum samples was established. The release of paclitaxel in vitro and the pharmacokinetics after i.v. bolus injection to mice were studied. RESULTS: The mean diameter of Brij78-SLN and F68-SLN is (103.5 +/- 29.2) nm and (220 +/- 98) nm, respectively. The nanoparticles release paclitaxel slowly and linearly, within 24 h, Brij78-SLN and F68-SLN release 8% and 20% of total drug, respectively. Long-circulation nanoparticles was found to stay in the blood circulation, with T 1/2 beta 10.1 h of F68-SLN, and T 1/2 beta 4.88 h of Brij78-SLN more than one commercialized paclitaxel injection, T 1/2 beta 1.3 h. CONCLUSION: Stearic acid might be a new drug carrier material in the future.

Glucose biosensing using glassy carbon electrode modified with polyhydroxy-C60, glucose oxidase and ionic-liquid.

Direct electrochemistry of glucose oxidase (GOD) was achieved when an ionic liquid/GOD-Polyhydroxy-C60 functional membrane was confined on a glassy carbon electrode (GCE). The cyclic voltammograms (CVs) of the modified GCE showed a pair of redox peaks with a formal potential (E°') of - 329 ± 2 mV. The heterogeneous electron transfer constant (k(s)) was 1.43 s-1. The modified GCE response to glucose was linear in the range from 0.02 to 2.0 mM. The detection limit was 1 µM. The apparent Michaelis-Menten constant (K(m)(app)) was 1.45 mM.