Conjugated microporous polymers-scaffolded enzyme cascade systems with enhanced catalytic activity

Enhancing catalytic activity of multi-enzyme in vitro through substrate channeling effect is promis-ing yet challenging.Herein,conjugated microporous polymers(CMPs)-scaffolded integrated en-zyme cascade systems(I-ECSs)are constructed through co-entrapping glucose oxidase(GOx)and horseradish peroxidase(HRP),in which hydrogen peroxide(H_(2)O_(2)) is the intermediate product.The interplay of low-resistance mass transfer pathway and appropriate pore wall-H_(2)O_(2) interactions facilitates the directed transfer of H_(2)O_(2),resulting in 2.4-fold and 5.0-fold elevation in catalytic activ-ity compared to free ECSs and separated ECSs,respectively.The substrate channeling effect could be regulated by altering the mass ratio of GOx to HRP.Besides,I-ECSs demonstrate excellent stabili-ties in harsh environments and multiple recycling.

Rat models of frozen shoulder:Classification and evaluation

Frozen shoulder(FS),also known as adhesive capsulitis,is a condition that causes contraction and stiffness of the shoulder joint capsule.The main symptoms are per-sistent shoulder pain and a limited range of motion in all directions.These symp-toms and poor prognosis affect people's physical health and quality of life.Currently,the specific mechanisms of FS remain unclear,and there is variability in treatment methods and their efficacy.Additionally,the early symptoms of FS are difficult to distinguish from those of other shoulder diseases,complicating early diagnosis and treatment.Therefore,it is necessary to develop and utilize animal models to under-stand the pathogenesis of FS and to explore treatment strategies,providing insights into the prevention and treatment of human FS.This paper reviews the rat models available for FS research,including external immobilization models,surgical internal immobilization models,injection modeling models,and endocrine modeling models.It introduces the basic procedures for these models and compares and analyzes the advantages,disadvantages,and applicability of each modeling method.Finally,our paper summarizes the common methods for evaluating FS rat models.

In-situ Leaching-immobilization of Lead in Soilvegetable System Based on Coupling of Remediation & Production

[Objective] The aim was to seek effective soil remediation technologies suitable for soils slightly or moderately polluted. [Method] In-situ leaching-immobi- lization was used to research remediation soil and vegetable contaminated by lead in vegetable bases in Chongqing. [Result] By tartaric acid-based leaching-sodium sulphide-based immobilization, vegetable Pb reduction rate was 46.4%, soil Pb reduction rate was 8.45% and Pb immobilization rate was 9.3% in deeper horizons. EDTA leaching did reduce lead content in vegetables, but reduced vegetable yield simultaneously. [Conclusion] The coupling remediation technology brought heavy metal contents down in vegetables without affecting productions. Therefore, it guarantees vegetable safety and reduces vegetable farmer＇s economic loss. It is a remediation technology suitable for soils slightly or moderately contaminated by heavy metals.

Culture of yeast cells immobilized by alginate-chitosan microcapsules in aqueous-organic solvent biphasic system

Immobilization biocatalysis is a potential technology to improve the activity and stability of biocatalysts in nonaqueous systems for efficient industrial production.Alginate-chitosan(AC)microcapsules were prepared as immobilization carriers by emulsifi cation-internal gelation and complexation reaction,and their contribution on facilitating the growth and metabolism of yeast cells were testifi ed successfully in culture medium-solvent biphasic systems.The cell growth in AC microcapsules is superior to that in alginate beads,and the cells in both immobilization carriers maintain much higher activity than free cells,which demonstrates AC microcapsules can confer yeast cells the ability to resist the adverse effect of solvent.Moreover,the performance of AC microcapsules in biphasic systems could be improved by adjusting the formation of outer polyelectrolyte complex(PEC)membrane to promote the cell growth and metabolic ability under the balance of resisting solvent toxicity and permitting substrate diffusion.Therefore,these findings are quite valuable for applying AC microcapsules as novel immobilization carriers to realize the biotransformation of value-added products in aqueous-solvent biphasic systems.

In-situ nitrogen removal from the eutrophic water by microbial-plant integrated system

Objective: This study was to assess the influence of interaction of combination of immobilized nitrogen cycling bacteria (INCB) with aquatic macrophytes on nitrogen removal from the eutrophic waterbody, and to get insight into different mechanisms involved in nitrogen removal. Methods: The aquatic macrophytes used include Eichhornia crassipes (sum-mer-autumn floating macrophyte), Elodea nuttallii (winter-growing submerged macrophyte), and nitrogen cycling bacteria in-cluding ammonifying, nitrosating, nitrifying and denitrifying bacteria isolated from Taihu Lake. The immobilization carriers materials were made from hydrophilic monomers 2-hydroxyethyl acrylate (HEA) and hydrophobic 2-hydroxyethyl methylacrylate (HEMA). Two experiments were conducted to evaluate the roles of macrophytes combined with INCB on nitrogen removal from eutrophic water during different seasons. Results: Eichhornia crassipes and Elodea nuttallii had different potentials in purification of eutrophic water. Floating macrophyte+bacteria (INCB) performed best in improving water quality (during the first experiment) and decreased total nitrogen (TN) by 70.2%, nitrite and ammonium by 92.2% and 50.9%, respectively, during the experimental period, when water transparency increased from 0.5 m to 1.8 m. When INCB was inoculated into the floating macrophyte system, the populations of nitrosating, nitrifying, and denitrifying bacteria increased by 1 to 2 orders of magnitude compared to the un-inoculated treatments, but ammonifying bacteria showed no obvious difference between different treatments. Lower values of chlorophyll a, CODMn, and pH were found in the microbial-plant integrated system, as compared to the control. Highest reduction in N was noted during the treatment with submerged macrophyte+INCB, being 26.1% for TN, 85.2% for nitrite, and 85.2% for ammonium at the end of 2nd experiment. And in the treatment, the populations of ammonifying, nitrosating, nitrifying, and de-nitrifying bacteria increased by 1 to 3 orders of magnitude, as compared to the un-inoculated treatments. Similar to the first ex-periment, higher water transparency and lower values of chlorophyll a, CODMn and pH were observed in the plant+ INCB inte-grated system, as compared to other treatments. These results indicated that plant-microbe interaction showed beneficial effects on N removal from the eutrophic waterbody.

Combination of adsorption and biodegradation processes for textile effluent treatment using a granular activated carbon-biofilm configured packed column system

The objective of this study was to investigate the feasibility of using a granular activated carbon-biofilm configured packed column system in the deeolodzation of azo dye Acid Orange 7-containing wastewater.The Acid Orange 7-degrading microbial from anaerobic sequencing batch reactor which treating the azo dye-containing wastewater for more than 200 d was immobilized on spent granular activated carbon(GAC)through attachment.The GAC-biofilm configured packed column system showed the ability to decolorize 10...

Optimization of Two-species Whole-cell Immobilization System Constructed with Marine-derived Fungi and Its Biological Degradation Ability

Mycelia pellet formed spontaneously in the process of cultivation was exploited as a biological carrier for whole-cell immobilization due to its unique structural characteristic. An innovative two-species whole-cell im- mobilization system was achieved by inoculating the marine-derived fungus Pestalotiopsis sp. J63 spores into cul- ture medium containing another fungus Penicillium janthinellum P1 pre-grown mycelia pellets for 2 days without any pretreatment. In order to evaluate the biological degradation capacity of this novel constructed immobilization system, the immobilized pellets were applied to treat paper mill effluent and decolorize dye Azure B. The use of the constructed immobilization system in the effluent resulted in successful and rapid biodegradation of numerous in- soluble fine fibers. The optimum conditions of immobilized procedure for maximum biodegradation capacity were determined using orthogonal design with biomass of P1 pellets 10 g （wet mass）, concentration of J63 spore 2x109 mlq, and immobilization time 2 d. The results demonstrate that immobilized pellets have more than 99% biodegradation capacity in a ten-hour treatment process. The kinetics of biodegradation fits the Michaelis-Menten equation well. Besides, the decolorization capability of immobilized pellets is more superior than that of P1 mycelia pellets. Overall, the present study offers a simple and reproducible way to construct a two-species whole-cell immobiliza- tion system for sewage treatment.

Study on Algae Removal by Immobilized Biosystem on Sponge

In this study, sponges were used to immobilize domesticated sludge microbes in a limited space, forming an immobilized biosystem capable of algae and microcystins removal. The removal effects on algae, microcystins and UV 260 of this biosystem and the mechanism of algae removal were studied. The results showed that active sludge from sewage treatment plants was able to remove algae from a eutrophic lake’s water after 7 d of domestication. The removal efficiency for algae, organic matter and microcystins increased when the domesticated sludge was immobilized on sponges. When the hydraulic retention time (HRT) was 5 h, the removal rates of algae, microcystins and UV 260 were 90%, 94.17% and 84%, respectively.The immobilized biosystem consisted mostly of bacteria, the Ciliata and Sarcodina protozoans and the Rotifer metazoans. Algal decomposition by zoogloea bacteria and preying by microcreatures were the two main modes of algal removal, which occurred in two steps: first, absorption by the zoogloea; second, decomposition by the zoogloea bacteria and the predacity of the microcreatures.

Asymmetric Synthesis of (-)-1-Trimethylsilyl-ethanol with Immobilized Saccharomyces Cerevisiae Cells in Water/Organic Solvent Diphasic System

Asymmetric synthesis of (-)-1-trimethylsilyl-ethanol with immobilized Saccharomyces cerevisiae cells in water/organic solvent biphasic system was studied. The effects of shake speed, hydrophobicity of organic solvent, volume ratio of water phase to organic phase, pH value of aqueous phase and reaction temperature on the initial reaction rate, maximum yield and enantiomeric excess (ee) of the product were systematically explored. All the above-mentioned factors had significant influence on the reaction. n-Hexane was found to be the best organic solvent for the reaction. The optimum shake speed, volume ratio of water phase to organic phase, pH value and reaction temperature were 150 r.min-1, 1/2, 8 and 30 ℃ respectively, under which the maximum yield and enantiomeric excess of the product were as high as 96.8% and 95.7%, which are 15% and 16% higher than those of the corresponding reaction performed in aqueous phase. To our best knowledge, this is the most satisfactory result obtained.

Phosphorus Bioavailability in a Phosphorus-Abundant System under Repeated Sediment Resuspension Conditions

Variation of bioavailable phosphorus(BAP) concentrations,i.e.,the sum of dissolved total phosphorus(DTP) and bioavailable particulate phosphorus(BAPP),was investigated in a phosphorus(P)-abundant system as influenced by repeated sediment resuspension.The overlying water and sediments in the studied system collected from a city canal,with the initial BAP concentration about 0.679 mg·L-1 in the overlying water.The sediments were disturbed with rotating blade once every day for 30 days in the experimental system.Concentrations of dissolved inorganic phosphorus(DIP),DTP,and total phosphorus(TP) in overlying water and algal available phosphorus(AAP),total phosphorus(Tot-P) in suspended particles were determined in diminishing frequency.It was found that the BAP decreased rapidly in the beginning few days in the sediment resuspension system,and then kept decreasing gradually at a low level,compared with the static conditions.This was mainly attributed to the sharp decrease of DIP.It is suggested that repeated sediment resuspension plays a significant role in phosphorus migration and converting it to more stable forms.

Immobilization of Laccase from Ganoderma lucidum LYL 263 in Sol-gel System

[Objective] The aim was to study the immobilization of laccase from Canoderma lucidum in three different sol-gel systems to find out the optimum ma- terials for laccase immobilization. [ Method ] Laccase from G. lucidum LYL 263 was immobilized in sodium alginate-gelatin, sodium alginate-chitosan and sodium alginate-gelatin-chitosan sol-gel systems, respectively; and the laccase activity was preliminarily analyzed. [ Result] The activity of laccase immobilized in sodium alginate-gelatin-chitosan was 2.14 folds and 2.75 folds respectively, compared to sodium alginate-gelatin and sodium alginate-chitosan; and this optimum system contained sodium alginate 2.0% , gelatin 1.0%, chitosan 0.3% and calcium chloride 56%. Benzoic acid was firstly used to embed immobilized laccase in this study, and single-factor experiments revealed that the activity of laccase could be effectively improved by suitable content of benzoic acid. The optimal conditions for laccase immobilization in the system were be^oie acid 2 retool/L, glutaraldehyde 0.32%, cross-linking reaction time 50 rain and enzyme concentration 10.0% ; and then the activity of the immobilized enzyme reached 635.7 U/g with these conditions. The thermal stability of the immobilized laccase was higher than free lac- case, and the activity of immobilized enzyme treated by freeze-drying was 1.59 folds compared to the non-freeze-dried, but the freeze-dried immobilized laccase had poor operational stability. [ Conclusion] The compositions of the material had significant influence on laccase immobilization and this study will provide reference to select materials for immobilization of laccase from G. lacidum LYL 263.

Strategy &Techniques for Restoration of Healthy Aquaecosystem from Toxic Super Eutrophic Water Body

There are complex river-lake systems in the Taihu Lake catchment with total water surface area of 6174.7 km2, and population density of 1079/km2, including Taihu Lake water surface area of 2338 km2. The water systems in this catchment have healthy aquaecosystems during long history. However, in some riverlets in this catchment the water quality was estimated as “acute toxicity for higher organisms” and over standards for many heavy metal elements content;there were no any living plants and macro organisms in the water body, because there were developed a series of industry with abundant release of heavy metals and difficult decomposition organic chemical components along the riverlets during last decades. The even more serious situation was observed in sediments of the riverlets. How to restore such riverlet into a healthy aquaeosystem with abound plants and higher organisms? The main strategy and techniques are described in this paper as summarizing a report of engineering in a riverlet in Wuxi New District during last years, which leads to restore the aquaecosystem into a healthy one with abundant surface plant cultured on floating islands and observed living fish, lobster, frog, toad, mollusk and others in the riverlet. The main techniques are: 1) softwall buffer technic;2) floating eco-island technic by using which can culture any plant which can be cultured in solution;3) immobilized nitrogen cycle bacteria (INCB) technic;4) tattering esters and other big-molecule organic chemicals by using electronic pulse technic and photosensitization technic;5) mist spray facility technic for improving dissolved oxygen in deep water layers;6) technic for buffering and suppressing H2S release from water;7) the appropriate portion of surface with cultured plant to the total water surface area is about 1/3;8) Cress [Oenanthe Ljavanica (Bl.) DC.] and Myriophyllum verticilatum L. may be cultured in Taihu Lake catchment during the whole year as main plants with mosaic combination of other supplement plants in different seasons.

Laccase Immobilized on Mesoporous Silica Materials and Its Corrosion Inhibition Performance in Circulating Cooling Water System

Mesoporous SiO_2 microspheres were synthesized using the sol-gel method and were characterized by TEM, FT-IR and BET techniques. The diameter of the microspheres is about 100—150 nm, and the average mesopore diameter is 2.55 nm, while the specific surface area is 1 088.9 m2/g. Mesoporous SiO_2 microspheres adsorb glutaraldehyde and immobilize laccase by means of the aldehyde group in glutaral which can react with the amidogen of laccase. The immobilization conditions were optimized at a glutaraldehyde concentration of 0.75%, a crosslinking time of 8 h, a laccase concentration of 0.04 L/L and an immobilization time of 10 h. When diesel leakage concentration was 80 mg/L, the highest corrosion inhibition efficiency of immobilized laccase reached 49.23%, which was slightly lower than the corrosion inhibition efficiency of free laccase(59%). The diesel degradation ratio could reach up to 45%. It has been proved that the immobilized laccase could degrade diesel to inhibit corrosion.

Synthesis of End Functional Polymers via Atom Transfer Radical Polymerization in Immobilized Catalytic System

Cross-linked polystyrene with azo-crown ether functional side chain (PSt-1, 10-dicarbonyl-3,6,9-trizaoeylcode-cane) was prepared under microwave irradiation and the structure was characterized through FT-IR and element analysis. The functionalized cross-linked polystyrene (cross-link degree, 3.5%) combining with immobilized catalyst system (CuBr and ethylα-bromo-isobutyrate) can catalyze atom transfer radical polymerization of Styrene. Neat polymer products can be obtained then. Complex of La and the polymer end group (EBiB) was synthesized. The third order nonlinear optical property of the polymer-La complex was investigated and the structure was also characterized by FT-IR and XPS.

Rational design of Aspergillus flavus A5p1-immobilized cell system to enhance the decolorization of reactive blue 4(RB4)

Anthraquinone dyes are a class of typical carcinogenic and hard-biodegradable organic pollutants.This study aimed to enhance the decolorization of anthraquinone dye by rationally designing an expected immobilized system.Reactive blue 4(RB4) was used as a substrate model and a previous isolated dyedegrading strain Aspergillus flavus A5pl was purposefully immobilized.Considering the effects of cell attachment and mass transfer,the polyurethane foam(PUF) with open pore structure was selected as the immobilization carrier.Results showed that the RB4 decolorization efficiency was significant enhanced after immobilization.Compared to the free mycelium system,the decolorization time of200 mg·L^(-1)RB4 was shortened from 48 h to 28 h by the PUF-immobilized cell system.Moreover,the PUF-immobilized system could tolerate RB4 up to 2000 mg-L^(-1).In the packed bed bioreactor(PBBR),an average decolorization efficiency of 93.3% could be maintained by the PUF-immobilized system for26 days.The decolorization process of RB4 was well described by the logistic equation and the degradation pathway was discussed.It was found that the higher specific growth rate of the PUF-immobilized cells was one of reasons for the enhanced decolorization.The good performance of the PUFimmobilized cell system would make it have potential application value for RB4 bioremediation.

Spine Stereotactic Body Radiation Therapy Residual Setup Errors and Intra-Fraction Motion Using the Stereotactic X-Ray Image Guidance Verification System

Purpose: To determine the precision of our institution’s current immobilization devices for spine SBRT, ultimately leading to recommendations for appropriate planning margins. Methods: We identified 12 patients (25 treatments) with spinal metastasis treated with spine Stereotactic Body Radiation Therapy (SBRT). The Body-FIX system was used as immobilization device for thoracic (T) and lumbar (L) spine lesions. The head and shoulder mask system was used as immobilization device for cervical (C) spine lesions. Initial patient setup used the infrared positioning system with body markers. Stereotactic X-ray imaging was then performed and correction was made if the initial setup error exceeded predetermined institutional tolerances, 1.5 mm for translation and 2° for rotation. Three additional sets of verification X-rays were obtained pre-, mid-, and post-treatment for all treatments. Results: Intrafraction motion regardless of immobilization technique was found to be 1.28 ± 0.57 mm. The mean and standard deviation of the variances along each direction were as follows: Superior-inferior, 0.56 ± 0.39 mm and 0.77 ± 0.52 mm, (p = 0.25);Anterior-posterior, 0.57 ± 0.43 mm and 1.14 ± 0.61 mm, (p = 0.01);Left-right, 0.48 ± 0.34 mm and 0.74 ± 0.40 mm, (p = 0.09) respectively. There was a significantly greater difference in the average 3D variance of the BodyFIX as compared to the head and shoulder mask immobilization system, 1.04 ± 0.46 mm and 1.71 ± 0.52 mm;(p = 0.003) respectively. Conclusions: Overall, our institution’s image guidance system using stereotactic X-ray imaging verification provides acceptable localization accuracy as previously defined in the literature. We observed a greater intrafraction motion for the head and shoulder mask as compared with the BodyFIX immobilization system, which may be a result of greater C-spine mobility and/or the suboptimal mask immobilization. Thus, better immobilization techniques for C-spine SBRT are needed to reduce setup error and intrafraction motion. We are currently exploring alternative C-spine immobilization techniques to improve set up accuracy and decrease intrafraction motion during treatment.

Analytical Expression for the Concentration of Substrate and Product in Immobilized Enzyme System in Biofuel/Biosensor

In this paper, an approximate analytical method to solve the non-linear differential equations in an immobilized enzyme film is presented. Analytical expressions for concentrations of substrate and product have been derived for all values of dimensionless parameter. Dimensionless numbers that can be used to study the effects of enzyme loading, enzymatic gel thickness, and oxidation/ reduction kinetics at the electrode in biosensor/biofuel cell performance were identified. Using the dimensionless numbers identified in this paper, and the plots representing the effects of these dimensionless numbers on concentrations and current in biosensor/biofuel cell are discussed. Analytical results are compared with simulation results and satisfactory agreement is noted.

Effectiveness of a Patient-Specific Immobilization and Positioning System to Limit Interfractional Translation and Rotation Setup Errors in Radiotherapy of Prostate Cancers

Objective: To evaluate the effectiveness of a patient-specific immobilization and positioning device in prostate radiotherapy. Methods: Eighty patients were immobilized and positioned by a patient-specific device. Prostate translations and rotations were estimated from daily cone beam computed tomography scans using a contour-based approach assisted by auto-registration and quantified by the group mean GM, systematic Σ and random σ' errors. Dosimetric impacts of residual prostate rotations where the translation errors were corrected were evaluated by robustness plan analysis. Results: Using the patient-specific immobilization alone without online image-guidance, the GM, Σ and σ' of the prostate translations were 0.8, 1.7, and 1.5 mm (left-right;LR), 0.8, 2.1, and 1.9 mm (superior-inferior;SI), and 0.5, 1.7 and 1.5 mm (anterior-posterior;AP), while for the prostate rotations they were 0.0°, 0.6°, and 0.7°(pitch), 0.2°, 0.5°, and 0.6°(roll), and 0.2°, 0.5°, and 0.6°(yaw). The resulting van Herk’s margin was 5.8 (LR), 7.3 (SI) and 5.8 (AP) mm. With adaptive online image-guidance based on estimates from the first 5 fractions, Σ were reduced by 0.7 - 1.2 mm for the prostate translations, resulting in a margin reduction by 2 - 3.5 mm. Changes of Σ and σ' in the prostate rotations were insignificant regardless of translation corrections. Dosimetric impacts of residual rotation errors were negligible if a 2 mm margin was applied. Conclusions: Our patient-specific immobilization system can effectively limit the prostate translations and rotations, which is important without 6D treatment couches or using ultrasound image-guidance without rotational corrections.

Quantitative measurement of p53-p53 antibody interactions by quartz crystal microbalance: A modelsystem for immunochemical calibration

We are developing methods to quantify antibody interactions that include a quartz crystal microbalance (QCM) system to measure, on a molecular basis, the interaction of p53 and anti-p53 antibodies. Previously, as a model system, we developed a measurement device consisting of p53 protein (human wild type), characterized by mass spectroscopy and immobilized at various concentrations on a glass slide. The device is designed as a control to be used with immunohistochemical (IHC) assays that incorporate commercially available anti-p53 antibodies and probes. In the current study, p53 protein is covalently immobilized on a silicon dioxide-coated quartz crystal and the resonance frequency shift is followed in-situ. The functionalized sensor is then incubated with the anti-p53 antibody, which provides a direct gravimetric measure of the antibody-antigen binding. This previously described method allows the comparison of the surface immobilized protein concentrations with estimates obtained by fluorescence measurement. The p53 functionalized QCM system offers an independent measure of surface immobilized protein concentration and is essential in development of our IHC calibration prototypes. These results provide a benchmark for comparing antibody systems that may be used in developing other molecular diagnostic assays such as immunocytochemical analysis and the detection of biomarker proteins in blood and urine.

STUDY ON IMMOBILIZED PORCINE PANCREATIC LIPASE CATALYZING TRANSESTERIFICATION BETWEEN METHYL－BUTYRATE AND 1－BUTANOL IN NONAQUEOUS SYSTEMS

Transesterification between methyl-butyrate and 1-butanol in nonaqueous systems was catalyzed by porcine pancreatic lipase which was immobilized on cross- linked polystyrene. Organic solvents, substrate concentration, contents of water and other parameters which affect the immobilized enzyme activity were studied. Lipase immobilized on hydrophobic crosslinked polystyrene can reduce its diffusion limit in the reaction. It was found that the activity of immobilized lipase in organic systems was two times as high as that of free lipase.