Kinetic method for enzymatic analysis by predicting background with uricase reaction as model

Objective：To investigate the reliability for kinetic assay of substance with background predicted by the integrated method using uricase reaction as model. Methods： Absorbance before uricase action （Δ0） was estimated by extrapolation with given lag time of steady-state reaction. With Km fixed at 12.5μmol/L, background absorbance （Δb） was predicted by nonlinearly fitting integrated Michaelis-Menten equation to Candida utilis uricase reaction curve. Uric acid in reaction solution was determined by the difference （ΔA） between Δ0 and Δb. Results .Ab usually showed deviation 〈3% from direct assay with residual substrate done fifth of initial substrate for analysis. ΔA showed CV 〈5% with resistance to common interferences except xanthine, and it linearly responded to uric acid with slope consistent to the absorptivity of uric acid. The lower limit was 2.0 μmol/L and upper limit reached 30 μmol/L in reaction solution with data monitored within 8 min reaction at 0. 015 U/ml uricase. Preliminary application to serum and urine gave better precision than the direct equilibrium method without the removal of proteins before analysis. Conclusion .This kinetic method with background predicted by the integrated method was reliable for enzymatic analysis, and it showed resistance to common interferences and enhanced efficiency at much lower cost.

A new enzymatic method for determination of E. coli

beta-Galactosidase, a kind of endoenzyme in E. coli cells. can be released by the pore-forming action of colicin El. and E. coli can be detected rapidly by enzymatic method.

Synthesis of CCK-8 Tetrapeptide Fragment by Enzymatic Method

The enzymatic synthesis of a tetrapeptide Phac-Met-Gly-Trp-Met-OEt, a fragment of the cholecystokinin C-terminal octapeptide CCK-8, was reported This fragment was synthesized by coupling Phac-Met-OEt with Gly-OMe, Trp-OMe and Met-OEt successively These three steps were catalyzed by α-chymotrpsin, Papain and α-chymotrpsin respectively The results of FAB-MS showed that all the products had the correct molecular mass

Analysis of Urea in Petfood Matrices: Comparison of Spectro-Colorimetric, Enzymatic and Liquid Chromatography Electrospray Ionization High Resolution Mass Spectrometry Methods

Adulteration may consist in non authorized source of nitrogen addition to increase the protein content of some raw materials. Urea which is authorized for feed is a non nutritional source of nitrogen in food and pet food. Adulteration of food or pet food raw material by urea is thus monitored by manufacturer and governmental authorities with official methods which are either enzymatic (Association of Official Agricultural Chemists, AOAC) or spectro-colorimetric (European Community, EC). Each method gives results which are not comparable and spectro-colorimetric methods may result in false-positive urea detection. Liquid chromatographic (LC/UV-DAD) analysis of extracts from spectro-colorimetric method indicates that presence of free amino-acid may interfere with colorimetric detection of urea in the EC method with pet food samples. Liquid chromatography electrospray ionization high resolution mass spectrometry (LC/ESI-HRMS) has allowed to quantify low content (<0.01%) of urea in pet food water extracts for samples which resulted in significant urea detection with colorimetric method and in content below the detection threshold with enzymatic method. This study demonstrates the EC colorimetric method is not applicable to pet food and also food samples which have a complex composition with significant levels of free amino acids. On the other hand we clearly evidenced by means of the LC/ESI-HRMS results that the AOC Enzymatic method is applicable to urea quantification in pet food samples and gives reliable results.

Aqueous Enzymatic Method to Extract Indigo Pigment from Baphicacanthus cusia Leaves

This study explored the extraction process of indigo pigment from Baphicacanthus cusia using aqueous enzymatic method for the first time. Through single factor tests and orthogonal tests, this study investigated the effects of extraction time, extraction temperature, enzyme concentration, pH value, and solidliquid ratio on the extraction rate of indigo pigment. The results showed that the extraction rate of indigo reached the highest(0.218%) when the solid-liquid ratio was 1:20, enzyme concentration was 6 g/L, pH value was 5, extraction temperature was 50℃, and extraction time was 3 h. The method had greatly shortened the extraction time, simplified the extraction process, and improved the extraction efficiency while producing little pollution and meeting the green environmental protection requirements.

Temperature-dependent photoluminescence on organic inorganic metal halide perovskite CH_3NH_3Pb I_(3-)Cl_ prepared on ZnO/FTO substrates using a two-step method

Temperature-dependent photoluminescence characteristics of organic-inorganic halide perovskite CH3NH3Pb I3-xClx films prepared using a two-step method on ZnO/FTO substrates were investigated. Surface morphology and absorption characteristics of the films were also studied. Scanning electron microscopy revealed large crystals and substrate coverage. The orthorhombic-to-tetragonal phase transition temperature was-140 K. The films＇ exciton binding energy was 77.6 ± 10.9 meV and the energy of optical phonons was 38.8 ± 2.5 meV. These results suggest that perovskite CH3NH3Pb I（3-x）Clx films have excellent optoelectronic characteristics which further suggests their potential usage in perovskitebased optoelectronic devices.

Applying the Dynamic Two-Step Method to Forecast Remaining Oil Distribution of Lower Series ,Xiaermen Oilfield

The distribution of remaining oil is often described qualitatively. The remaining oil distributed in the whole reservoir is calculated according to the characteristics of the space distribution of the saturation of remaining oil. Logging data are required to accomplish this. However, many such projects cannot be completed. Since the old study of remaining oil distribution could not be quantified efficiently, the ＂dynamic two-step method＂ is presented. Firstly, the water cut of every flow unit in one well at one time is calculated according to the comprehensive water cut of a single well at one time. Secondly, the remaining oil saturation of the flow unit of the well at one time is calculated based on the water cut of the flow unit at a given time. The results show that ＂dynamic two-step method＂ has characteristics of simplicity and convenience, and is especially suitable for the study of remaining oil distribution at high water-cut stage. The distribution of remaining oil presented banding and potato form, remaining oil was relatively concentrated in faultage neighborhood and imperfect well netting position, and the net thickness of the place was great. This proposal can provide an effective way to forecast remaining oil distribution and enhance oil recovery, especially applied at the high water-cut stage.

Enzymatic Aqueous Extraction of Soybean Oil

The procedure of enzymatic aqueous extraction of soybean oil was assessed when two-step controlled enzymatic hydrolysis was applied. With aqueous extraction of soybean oil-containing protein, the highest yield of oil was 96.1% at the optimized conditions studied. Soybean oil-containing protein was hydrolyzed and resulted in releasing part of oil. The separated protein that contained 40% oil was enriched due to its adsorption capacity of released oil, the average oil extraction yeild reached 93.5%. Then the high oil content protein was hydrolyzed again to release oil by enzyme, the oil extraction yeild was 80.4%. As a result, high quality of soybean oil was obtained and the content of total oil yield was 74.4%.

Enzymatic Synthesis of a CCK-8 Tetrapeptide Fragment

The enzymatic synthesis of a tetrapeptide Phac-Met-Gly-Trp-Met-OEt is reported. It was synthesized by coupling Phac-Met-OEt with Gly-OMe ·HC1, Trp-OMe and Met-OEt successively, catalyzed by α-chymotrypsin, papain and α-chymotrypsin respectively. The results of FAB-MS showed that the products had the correct molecular mass.

The convergence ball and error analysis of the two-step Secant method

Under the assumption that the nonlinear operator has Lipschitz continuous divided differences for the first order,we obtain an estimate of the radius of the convergence ball for the two-step secant method.Moreover,we also provide an error estimate that matches the convergence order of the two-step secant method.At last,we give an application of the proposed theorem.

A new two-step variational model for multiplicative noise removal with applications to texture images

Multiplicative noise removal problems have attracted much attention in recent years.Unlike additive noise,multiplicative noise destroys almost all information of the original image,especially for texture images.Motivated by the TV-Stokes model,we propose a new two-step variational model to denoise the texture images corrupted by multiplicative noise with a good geometry explanation in this paper.In the first step,we convert the multiplicative denoising problem into an additive one by the logarithm transform and propagate the isophote directions in the tangential field smoothing.Once the isophote directions are constructed,an image is restored to fit the constructed directions in the second step.The existence and uniqueness of the solution to the variational problems are proved.In these two steps,we use the gradient descent method and construct finite difference schemes to solve the problems.Especially,the augmented Lagrangian method and the fast Fourier transform are adopted to accelerate the calculation.Experimental results show that the proposed model can remove the multiplicative noise efficiently and protect the texture well.

Solution of Nonlinear Integro Differential Equations by Two-Step Adomian Decomposition Method (TSAM)

The Adomian decomposition method (ADM) can be used to solve a wide range of problems and usually gets the solution in a series form. In this paper, we propose two-step Adomian Decomposition Method (TSAM) for nonlinear integro-differential equations that will facilitate the calculations. In this modification, compared to the standard Adomian decomposition method, the size of calculations was reduced. This modification also avoids computing Adomian polynomials. Numerical results are given to show the efficiency and performance of this method.

Arrhenius relationship and two-step scheme in AF hyperdynamics simulation of diffusion of Mg/Zn interface

The accelerating factor （AF） method is a simple and appropriate way to investigate the atomic long-time deep diffusion at solid-solid interface. In the framework of AF hyperdynamics （HD） simulation, the relationship between the diffusion coefficient along the direction of z-axis which is normal to the Mg/Zn interface and temperature was investigated, and the AF＇s impact on the diffusion constant （D0） and activation energy （Q^＊） was studied. Then, two steps were taken to simulate the atomic diffusion process and the formation of new phases： one for acceleration and the other for equilibration. The results show that： the Arrhenius equation works well for the description of Dz with different accelerating factors; the AF has no effect on the diffusion constant Do in the case of no phase transition; and the relationship between Q＊ and Q conforms to Q^＊=Q/A. Then, the new Arrhenius equation for AFHD is successfully constructed as Dz=Doexp[-Q/（ART）]. Meanwhile, the authentic equilibrium conformations at any dynamic moment can only be reproduced by the equilibration simulation of the HD-simulated configurations. Key words： accelerating factor method; Arrhenius equation; two-steps scheme; Mg/Zn interface; hyperdynamic simulation

A Novel Method of Fabricating a Well-Faceted Large-Crystal Diamond Through MPCVD

A novel method was developed to deposit a large crystal diamond with good facets up to 1000 μm on a tungsten substrate using a microwave plasma enhanced chemical vapor deposition （MPCVD）. This method consists of two steps, namely single-crystal nucleation and growth. Prior to the fabrication of the well-faceted, large crystal diamond, an investigation was made into the nucleation and growth of the diamond which were affected by the O2 concentration and substrate temperature. Deposited diamond crystals were characterized by scanning electron microscopy and micro-Raman spectroscopy. The results showed that the conditions of single-crystal nucleation were appropriate when the ratio of H2/CH4/O2 was about 200/7.0/2.0, while the sub- strate temperature Ts of 1000℃ to 1050℃ was the appropriate range for single-crystal diamond growth. Under the optimum parameters, a well-faeeted large crystal diamond was obtained.

The Research on Biogas Power Generation by Two Step Method

The paper aims to demonstrate the system of SANYI biogas power generation by two-step method which can produce methane and generate electricity effectively and stably.So it can be stable synchronization meshwork energy source establishment and provide references to develop an effective and stable project of producing methane and generating electricity.

In-depth understanding of ionic liquid assisted perovskite film formation mechanism for two-step perovskite photovoltaics

Ionic liquids(ILs)have been widely applied in the one-step fabrication of perovskite with noticeable enhancement in the device performance.However,in-depth mechanism of ionic-liquid-assisted perovskite film formation is not well understood for also important two-step perovskite fabrication method,with better control of crystallization behavior.In this work,we introduced ionic liquid methylammonium formate(MAFa)into organic salt to produce perovskite film via a two-step method.Systematic investigations on the influence of MAFa on the perovskite thin film formation mechanism were performed.Ionic liquid is shown to assist lowering the perovskite formation enthalpy upon the density functional theory(DFT)calculation,leading to an accelerated crystallization process evidenced by in-situ UV-Vis absorption measurement.A gradient up-down distribution of ionic liquid has been confirmed by timeof-flight SIMS.Importantly,besides the surface passivation,we found the HCOO-can diffuse into the perovskite crystals to fill up the halide vacancies,resulting in significant reduction of trap states.Uniform perovskite films with significantly larger grains and less defect density were prepared with the help of MAFa IL,and the corresponding device efficiency over 23%was obtained by two-step process with remarkably improved stability.This research work provides an efficient strategy to tune the morphology and opto-electronic properties of perovskite materials via ionic-liquid-assisted two-step fabrication method,which is beneficial for upscaling and application of perovskite photovoltaics.

A class of twostep continuity Runge-Kutta methods for solving singular delay differential equations and its convergence

An idea of relaxing the effect of delay when computing the Runge-Kutta stages in the current step and a class of two-step continuity Runge-Kutta methods （TSCRK） is presented. Their construction, their order conditions and their convergence are studied. The two-step continuity Runge-Kutta methods possess good numerical stability properties and higher stage-order, and keep the explicit process of computing the Runge-Kutta stages. The numerical experiments show that the TSCRK methods are efficient.

Technology of Extracting Kiwi Fruit Seed Oil with Ultrasonic-Assisted Enzyme and Response Surface Method

Taken kiwi fruit as raw material, this paper extracted kiwi fruit seed oil with ultrasonic-assisted enzyme, researched the influence of factors such as liquid-to-solid ratio, granularity, type of enzyme, ultrasonic power, treating time, enzymolysis temperature, enzymolysis time, pH and enzyme additive on oil extraction, and optimized the extracting technology of kiwi fruit seed oil with response surface method. The result shows that the best technical parameter is: material granularity: 60, liquid-to-solid ratio: 1:10 (g/mL), ultrasonic power: 400 W, treating time: 30 min, enzyme amount: 2.50%, pH: 9.2, enzymolysis temperature: 53&deg;C, enzymolysis time: 2.80 h;and the extracting ratio under such condition is 92.57%.

A Remarkable Chord Iterative Method for Roots of Uncertain Multiplicity

In this note we at first briefly review iterative methods for effectively approaching a root of an unknown multiplicity. We describe a first order, then a second order estimate for the multiplicity index m of the approached root. Next we present a second order, two-step method for iteratively nearing a root of an unknown multiplicity. Subsequently, we introduce a novel chord, or a two- step method, not requiring beforehand knowledge of the multiplicity index m of the sought root, nor requiring higher order derivatives of the equilibrium function, which is quadratically convergent for any , and then reverts to superlinear.

Impact of creatinine methodology on glomerular filtration rate estimation in diabetes

AIM To evaluate the influence of creatinine methodology on the performance of chronic kidney disease(CKD)-Epidemiology Collaboration Group-calculated estimated glomerular filtration rate(CKD-EPI-eGFR) for CKD diagnosis/staging in a large cohort of diabetic patients. METHODS Fasting blood samples were taken from diabetic patients attending our clinic for their regular annual examination, including laboratory measurement of serum creatinine and eGFR.RESULTS Our results indicated an overall excellent agreement in CKD staging(kappa = 0.918) between the Jaffé serum creatinine-and enzymatic serum creatinine-based CKDEPI-eGFR, with 9% of discordant cases. As compared to the enzymatic creatinine, the majority of discordances(8%) were positive, i.e., associated with the more advanced CKD stage re-classification, whereas only 1% of cases were negatively discordant if Jaffé creatinine was used for eGFR calculation. A minor proportion of the discordant cases(3.5%) were re-classified into clinically relevant CKD stage indicating mildly to moderately decreased kidney function(< 60 m L/min per 1.73 m^2). Significant acute and chronic hyperglycaemia, assessedas plasma glucose and Hb A1 c levels far above the recommended glycaemic goals, was associated with positively discordant cases. Due to a very low frequency, positive discordance is not likely to present a great burden for the health-care providers, while intensified medical care may actually be beneficial for the small number of discordant patients. On the other hand, a very low proportion of negatively discordant cases(1%) at the 60 m L/min per 1.73 m^2 eGFR level indicate a negligible possibility to miss the CKD diagnosis, which could be the most prominent clinical problem affecting patient care, considering high risk of CKD for adverse patient outcomes. CONCLUSION This study indicate that compensated Jaffé creatinine procedure, in spite of the glucose-dependent bias, is not inferior to enzymatic creatinine in CKD diagnosis/staging and therefore may provide a reliable and cost-effective tool for the renal function assessment in diabetic patients.