Optimization of acidic extraction of astaxanthin from Phaffia rhodozyma

Optimization of a process for extracting astaxanthin from Phaffia rhodozyma by acidic method was investigated,regarding several extraction factors such as acids, organic solvents, temperature and time. Fractional factorial design, central composite design and response surface methodology were used to derive a statistically optimal model, which corresponded to the following optimal condition concentration of lactic acid at 5.55 mol/L, ratio of ethanol to yeast dry weight at 20.25 ml/g, temperature for cell-disruption at 30 ℃, and extraction time for 3 min. Under this condition, astaxanthin and the total carotenoids could be extracted in amounts of 1294.7 μg/g and 1516.0 μg/g, respectively. This acidic method has advantages such as high extraction efficiency, low chemical toxicity and no special requirement of instruments. Therefore, it might be a more feasible and practical method for industrial practice.     

Studies on optimization of nitrogen sources for astaxanthin production by <Emphasis Type="Italic">Phaffia rhodozyma</Emphasis>

Fermentation ofPhaffia rhodozyma is a major method for producing astaxanthin, an important pigment with industrial and pharmaceutical application. To improve astaxanthin productivity, single factor and mixture design experiments were used to investigate the effects of nitrogen source on Phaffia rhodozyma cultivation and astaxanthin production. Results of single factor experiments showed nitrogen source could significantly affect P. rhodozyma cultivation with respect to carbon source utilization, yeast growth and astaxanthin accumulation. Further studies of mixture design experiments using （NH4）2SO4, KNO3 and beef extract as nitrogen sources indicated that the proportion of three nitrogen sources was very important to astaxanthin production. Validation experiments showed that the optimal nitrogen source was composed of 0.28 g/L （NH4）2SO4, 0.49 g/L KNO3 and 1.19 g/L beef extract. The kinetic characteristics of batch cultivation were investigated in a 5-L pH-stat fermentor. The maximum amount of biomass and highest astaxanthin yield in terms of volume and in terms of biomass were 7.71 mg/L and 1.00 mg/g, respectively.     

Development and evaluation of immunoassay for zeranol in bovine urine

A high affinity polyclonal antibody-based enzyme linked immunosorbent assay （ELISA） was developed for the quantification of zeranol in bovine urine. On the basis of urine matrix studies, the optimized dilution factors producing insignificant matrix interference were selected as 1：5 in pretreatment. In the improved ELISA, the linear response range was between 0.02 and 1 μg/ml, and the detection limit was 0.02 μg/ml for the assay. The overall recoveries and the coefficients of variation （CVs） were in the range of 82%-127% and 3.5%-8.8%, respectively. Thirty-six bovine urine samples spiked with zeranol （ranging from 0.2 to 10 μg/ml） were detected by the ELISA and liquid chromatography （LC） method, and good correlations were obtained between the two methods （R^2=0.9643）. We conclude that this improved ELISA is suitable tool for a mass zeranol screening and can be an altemative for the conventional LC method for zeranol in bovine urine.     

Surface Modification of Cu-Cr Complex by NIR and MIR Laser简

Irradiated by infrared laser, the surface reducibility and adsorbability of Cu-Cr complex could be improved, owing to the interaction of photo-fragmentation and laser texturing. Analyzed by the binding energy spectra and the auger spectra, the valence states of chromium ion and copper ion were+3 and+1 after radiation respectively, which still had the reducibility to release electrons. In contrast with the near-infrared(NIR)1 064 nm and mid-infrared(MIR) 10 600 nm laser at the same average output power of 15 W, the reduced metal percentage in the Cu-Cr complex was obviously distinguished at the depth from nanometer to micron. After chemical plating, the average coating thickness and mean-square deviation of the NIR sample were 11.61 μm and 0.30 for copper layer, and 2.69 μm and 0.08 for nickel layer. The results were much better than those of the MIR sample.     

Development of an indirect competitive ELISA for simultaneous detection of enrofloxacin and ciprofloxacin

Modified 1-ethyl-3-(3-dimethylaminopropy) carbodiimide(EDC) method was employed to synthesize the artificial antigen of enrofloxacin(ENR),and New Zealand rabbits were used to produce anti-ENR polyclonal antibody(pAb).Based on the checkerboard titration,an indirect competitive enzyme-linked immunosorbent assay(ELISA) standard curve was established.This assay was sensitive and had a linear range from 0.6 to 148.0 μg/kg(R2 =0.9567),with the half maximal inhibitory concentration(IC50) and limit of detection(LOD) values of 9.4 μg/kg and 0.2 μg/kg,respectively.Of all the competitive analogues,the produced pAb exhibited a high cross-reactivity to ciprofloxacin(CIP)(87%),the main metabolite of ENR in tissues.After optimization,the matrix effects can be ignored using a 10-fold dilution in beef and 20-fold dilution in pork.The overall recoveries and coefficients of variation(CVs) were in the ranges of 86%-109% and 6.8%-13.1%,respectively.It can be concluded that the established ELISA method is suitable for simultaneous detection of ENR and CIP in animal tissues.     

Comparison Between MIR and NIR Spectroscopic Techniques for the Determination of Fat and Protein Contents in Milk

To compare mid-infrared(MIR)and near-infrared(NIR)spectroscopies for the determination of the fat and protein contents in milk,the same sample sets with varying concentrations of fat and protein were measured in the MIR range of 3 200-700 cm-1 and NIR range of 9 000-4 000 cm-1.The spectral features in the two regions were analyzed.The MIR spectra of milk were characteristic due to the MIR inherent molecular specificity,whereas the NIR spectra were relatively characterless due to the NIR low selectivity.Partial least squares(PLS)regression models for fat and protein were developed by using both MIR and NIR spectra.MIR data with no pretreatment gave better results than NIR data.The square correlation coefficient(R2)and the root mean square error of prediction(RMSEP)were 0.98 and 0.10 g/dL for fat and 0.97 and 0.11 g/dL for protein.With NIR techniques,satisfactory results were not obtained with raw data.However,NIR data after pretreatment gave similarly good results to the ones using MIR method.This paper indicates that either of the MIR and NIR spectral methods is reliable for the determination of the fat and protein contents.     

Inductorless CMOS Low Noise Amplifier for Multiband Application in 0.1–1.2 GHz

A 0.18 μm CMOS low noise amplifier(LNA) by utilizing noise-canceling technique was designed and implemented in this paper. Current-reuse and self-bias techniques were used in the first stage to achieve input matching and reduce power consumption. The core size of the proposed CMOS LNA circuit without inductor was only 128 μm 9226 μm. The measured power gain and noise figure of the proposed LNA were 20.6 and 1.9 dB,respectively. The 3-dB bandwidth covers frequency from 0.1 to 1.2 GHz. When the chip was operated at a supply voltage of 1.8 V, it consumed 25.69 mW. The high performance of the proposed LNA makes it suitable for multistandard low-cost receiver front-ends within the above frequency range.     

Prediction of valid acidity in intact apples with Fourier transform near infrared spectroscopy

To develop nondestructive acidity prediction for intact Fuji apples, the potential of Fourier transform near infrared (FT-NIR) method with fiber optics in interactance mode was investigated. Interactance in the 800 nm to 2619 nm region was measured for intact apples, harvested from early to late maturity stages. Spectral data were analyzed by two multivariate calibration techniques including partial least squares (PLS) and principal component regression (PCR) methods. A total of 120 Fuji apples were tested and 80 of them were used to form a calibration data set. The influences of different data preprocessing and spectra treatments were also quantified. Calibration models based on smoothing spectra were slightly worse than that based on derivative spectra, and the best result was obtained when the segment length was 5 nm and the gap size was 10 points. Depending on data preprocessing and PLS method, the best prediction model yielded correlation coefficient of determination (r^2) of 0.759,low root mean square error of prediction (RMSEP) of 0.0677, low root mean square error of calibration (RMSEC) of 0.0562. The results indicated the feasibility of FT-NIR spectral analysis for predicting apple valid acidity in a nondestructive way.     

Application of land use regression for estimating concentrations of major outdoor air pollutants in Jinan, China

SO2, NO2, and PM10 are the major outdoor air pollutants in China, and most of the cities in China have regulatory monitoring sites for these three air pollutants. In this study, we developed a land use regression （LUR） model using regulatory monitoring data to predict the spatial distribution of air pollutant concentrations in Jinan, China. Traffic, land use and census data, and meteorological and physical conditions were included as candidate independent variables, and were tabulated for buffers of varying radii. SO2, NO2, and PM10 concentrations were most highly correlated with the area of industrial land within a buffer of 0.5 km （R2=0.34）, the distance from an expressway （R2=0.45）, and the area of residential land within a buffer of 1.5 km （R2=0.25）, respectively. Three multiple linear regression （MLR） equations were established based on the most significant variables （p〈0.05） for SO2, NO2, and PM10, and R2 values obtained were 0.617, 0.640, and 0.600, respectively. An LUR model can be applied to an area with complex terrain. The buffer radii of independent variables for SO2, NO2, and PM10 were chosen to be 0.5, 2, and 1.5 kin, respectively based on univariate models. Intercepts of MLR equations can reflect the background concentrations in a certain area, but in this study the intercept values seemed to be higher than background concentrations. Most of the cities in China have a network of regulatory monitoring sites. However, the number of sites has been limited by the level of financial support available. The results of this study could be helpful in promoting the application of LUR models for monitoring pollutants in Chinese cities.     

Simultaneous determination of saponins in Radix Glycyrrhizae, Notoginseng and Ginseng by high performance liquid chromatography

To establish a method for determining five saponins（notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and ammonium glycyrrhizinate） in Glycyrrhizae, Notoginseng and Ginseng, the high performance liquid chromatography with diode array detector（HPLC-DAD） method was applied to an Inertsil ODS-SP column（4.6 mm×250 mm, 5 μm） with a mobile phase consisting of acetonitrile-0.05% phosphoric acid in a gradient elution manner. The flow rate was 1.0 mL/min. The column temperature was 30 ℃ and the detection wavelengths were 203 nm and 237 nm, respectively. The linear ranges were 0.700,0—7.000,0 μg for R1（r=1.000,0）, 0.751,1— 7.511,4 μg for Rg1（r=1.000,0）, 0.677,2—6.771,6 μg for Re（r=1.000,0）, 0.733,9—7.339,1 μg for Rb1（r= 1.000,0）, and 0.540,0—5.399,8 μg for ammonium glycyrrhizinate（r=0.999,9）, respectively. In addition, their average recoveries were 100.28%, 105.83%, 104.09%, 99.36% and 98.54%, respectively. The relative standard deviations（RSDs） of precision, reproducibility and recovery were all less than 1.5%. The results indicate that the method is simple, accurate and reproducible so that it can be used for the simultaneous determination of the five saponins in Chinese patent medicines containing the three kinds of herbs.     

Measurement and analysis of soil nitrogen and organic matter content using near-infrared spectroscopy techniques

Near infrared reflectance （N1R） spectroscopy is as a rapid, convenient and simple nondestructive technique useful for quantifying several soil properties. This method was used to estimate nitrogen （N） and organic matter （OM） content in a soil of Zhejiang Province, Hangzhou County. A total of 125 soil samples were taken from the field. Ninety-five samples spectra were used during the calibration and cross validation stage. Thirty samples spectra were used to predict N and OM concentration. NIR spectra of these samples were correlated using partial least square regression. The regression coefficients between measured and predicted values of N and OM was 0.92 and 0.93, and SEP （standard error of prediction） were 3.28 and 0.06, respectively, which showed that NIR method had potential to accurately predict these constituents in this soil. The results showed that NIR spectroscopy could be a good tool for precision farming application.     

Oxidative stress and damage induced by abnormal free radical reactions and IgA nephropathy

Objective: To estimate the oxidative stress and oxidative damage induced by abnormal free radical reactions in IgA nephropathy (IgAN) patients' bodies. Methods: Seventy-two IgA N patients (IgANP) and 72 healthy adult volunteers (HAV) were enrolled in a random control study design, in which the levels of nitric oxide (NO) in plasma, lipoperoxide (LPO) in plasma and in erythrocytes, and vitamin C (VC), vitamin E (VE) and β-carotene (β-CAR) in plasma as well as the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) in erythrocytes were determined with spectrophotometric mothods. Results: Compared with the HAV group, the averages of NO in plasma, and LPO in plasma and in erythrocytes in the IgANP group were significantly increased (P＜0.0001), while those ofVC, VE and β-CAR in plasma as well as those of SOD, CAT and GPX in erythrocytes in the IgANP group were significantly decreased (P＜0.0001). Linear correlation analysis showed that with the increase of the values of NO, and LPO in plasma and in erythrocytes, and with the decrease of those ofVC, VE, β-CAR,SOD, CAT and GPX in the IgAN patients, the degree of histological damage of tubulointerstitial regions was increased gradually (P＜0.0001); and that with the prolongation of the duration of disease the values of NO, and LPO in plasma and erythrocytes were increased gradually, while those of VC, VE, β-CAR, SOD, CAT and GPX were decreased gradually (P＜0.005). The discriminatory correct rates of the above biochemical parameters reflecting oxidative damage of the IgAN patients were 73.8%-92.5%, and the correct rates for the HAV were 70.0%-91.3% when independent discriminant analysis was used; and the correct rate for the IgAN patients was increased to 98.8%, the correct rate for the HAV was increased to 100% when stepwise discriminant analysis was used. The above biochemical parameters' reliability coefficient (alpha) were used to estimate the oxidative damage of the IgAN patients as 0.8145, the standardized item alpha=0.9730, F=53273.5681, P＜0.0001. Conclusions: A series of free radical chain reactions caused serious pathological aggravation in the IgANP' bodies, thus resulting in oxidative damage in their bodies. In treating IgANP, therefore, it is necessary that suitable dose antioxidants should be supplemented to them so as to alleviate the oxidative damage in their bodies.     

Application of the ISCST3 model for predicting PCDD/F concentrations in agricultural soil in the vicinity of a MSWI plant in China

Based on the Industrial Source Complex Short-Term Version 3 （ISCST3） model, a simplified modeling approach was developed to predict concentrations of congeners of polychlorinated-p-dioxins and dibenzofurans （PCDD/Fs） of agricultural soil, within a radius of 3 kin from a municipal solid waste incinerator （MSWI） plant after its 4-year operation in Hangzhou, China. Comparisons were made between the measured and estimated congener-specific concentrations and the international-toxic equivalent （I-TEQ） values of soil samples with respect to distance from the stack. The results indicate that the predictions of soil PCDD/F concentrations and K-TEQ values were generally lower than their observations, and that the higher the degree of underestimation seems, the greater the further downwind one gets. Nevertheless, most of the predictions were in good agreement with the trend of measured ones and were within a factor of ten for samples located within 1 kin of the plant. Besides, analysis of contributions of various deposition pathways confirms that in addition to wet particle deposition, the dry gaseous deposition is essential for realistic prediction of PCDD/F depositions to soil, especially for tetra- and penta-chlorinated dioxins.     

Seismic ductility of very-high-strength-concrete short columns subject to combined axial loading and cyclic lateral loading

The seismic ductility of reinforced very-high-strength-concrete (VHSC) short columns was studied by combinatively applying axial load and low cyclic lateral load on specimens to simulate seismic impact. Twelve specimens with concrete compressive strength ranging from 95.6 MPa to 118.6 MPa and a shear-span ratio of 2.0 were tested for shear failure pattern and fear force-displacement hysteretic responses. Combinative application of axial load and low cyclic lateral load to VHSC short columns incurs shear failure. The displacement ductility is much smaller when the axial load ratio is larger;whereas a larger stirrup ratio is accompanied with a better displacement ductility. The relationship of displacement ductility factor, μ?, with stirrup characteristic value, λv, and test axial load ratio, nt, is μ?=(1 8λv)/(0.33 nt). By this relationship and relevant codes for aseismatic design, the axial load ratio limits for aseismatic design of reinforced VHSC (C95 to C100) short columns for frame construction are respectively 0.5, 0.6, and 0.7 for seismic classes I, II, and Ⅲ;corresponding minimum characteristic values of stirrups are calculated according to the required characteristic values of at least 1.273 times of experimental results. These data are very useful to aseismatic engineering.     

Determination of benzalkonium chloride in viscous ophthalmic drops of azithromycin by high-performance liquid chromatography

A high-performance liquid chromatography (HPLC) system was used in the reversed phase mode for the determination of benzalkonium chloride (BKC) in azithromycin viscous ophthalmic drops. A Venusil-XBP(L)-C_(18) (150 mm×4.6 mm, 5 μm) column was used at 50 ℃. The mobile phase consisted of a mixture of methanol-potassium phosphate (16:5, v/v). Two sample preparation methods were compared. The results suggested that, compared with an extraction procedure, a deproteinization procedure was much quicker and more convenient. Using the deproteinization procedure for sample preparation, calibration curves were linear in the range 5.0～50 μg/ml. The within-day and inter-day coefficients of variation were less than 10%. The average recoveries were determined as 96.70%, 98.52%, and 97.96% at concentrations of 10.0, 30.0, and 50.0 μg/ml, respectively. Variability in precision did not exceed 5%. In conclusion, this HPLC method using a simple sample treatment procedure appears suitable for monitoring BKC content in azithromycin viscous ophthalmic drops.     

Ultimate stress increase in unbonded tendons in prestressed concrete beams

Since the assumption of plane sections cannot be applied to the strain of unbonded tendons in prestressed concrete beams subjected to loadings,a moment-curvature nonlinear analysis method is used to develop analytical programs from stress increases in unbonded tendons at the ultimate limit state.Based on the results of model testing and simulation analysis,equations are proposed to predict the stress increase in tendons at the ultimate state in simple or continuous beams of partially prestressed concrete,considering the loading type,non-prestressed reinforcement index βp,prestressing reinforcement index βs,and span-depth ratio L/h as the basic parameters.Results of 380 beams studied here and test results for 35 simple beams obtained by the China Academy of Building Research were compared with those from prediction equations given in codes and other previous studies.The comparison reveals that the values predicted by the proposed equations agree well with experimental results.     

Preparation of Glutaraldehyde Cross-Linked Chitosan Beads Under Microwave Irradiation and Properties of Urease Immobilized onto the Beads

The glutaraldehyde cross-linked chitosan beads were prepared under microwave irradiation and urease was immobilized onto the beads. The activity and the yield of enzyme activity of the immobilized urease were 10.83 U/g carrier and 47.7%, respectively. The optimum conditions of immobilization were 1% of glutaraldehyde volume fraction, 10 mg/g of urease/beads weight ratio, 24 h of the processing time and pH 6.5 of the reaction medium for immobilization. The properties of the immobilized urease were investigated and compared with those of the free enzyme. The optimum pH values were 6.5 and 7.0 for the immobilized and free urease, respectively. The optimum temperature was 60℃ for the free urease, while it shifted to 65 ℃ for the immobilized enzyme. The Michaelis constant Kr, was 9.1 mmol/L for the immobilized and 12.5 mmol/L for the free urease. The immobilized urease retained 40% of its initial enzyme activity even after 10 repeated uses. The immobilized urease stored at 4 ℃ retained 46% of its initial activity even after 35 d.     

Wetland bacteria isolated from Huangpu River-Yangtze River estuary and its degradation on diesel

Oil contaminated soil was collected from Huangpu River-Yangtze River estuary wetland, with the aim of isolating oil-degrading microorganisms and evaluating their ability to degrade diesel. Three bacterial strains were discovered and identified by sequencing their 16S rDNA genes, two were Pseudomonas and one was Alcaligcnes. The proper growth conditions of each bacterium were measured and presented for diesel biodegradation. Biodegradation assays revealed that the degradation rates of three bacterial strains were 42.5%, 14.6% and 15.9% in 7 d respectively. They all play an important role on the nalkanes within the range of C16-C25 components of diesel. The results indicated that the oil-degraders can adapt to degrade diesel. The bacterial strains can be used in wetland diesel pollution control.     

Determination of accurate theoretical values for thermodynamic properties in bulk metallic glasses

Deviation values of specific heat difference ACp, the Gibbs free energy difference AG, enthalpy dif- ference AH, and entropy difference AS between the supercooled liquid and corresponding crystalline phase produced by the linear, hyperbolic, and Dubey＇s expres- sions of ACp and the corresponding experimental values are determined for sixteen bulk metallic glasses （BMGs） from the glass transition temperature Tg to the melting temperature Tm. The calculated values produced by the hyperbolic expression for ACp most closely approximate experimental values, indicating that the hyperbolic ACp expression can be considered universally applicable, compared to linear and Dubey＇s expressions for ACp, which are accurate only within a limited range of condi- tions. For instance, Dubey＇s ACp expression provides a good approximation of actual experimental values within certain conditions （i.e., ~ = ACg/AC~ 〈 2, where ACpg and ACp represent the specific heat difference at temperatures Tg and Tm, respectively）.     

球磨工艺对活性炭孔结构及电化学性能的影响

The effects of particle size of activated carbon （AC） on its wettability, electrode coating technology and electric chemical performance were studied to assemble nonaqueous electric double layer capacitors （EDLCs） for high power requirements. The results show that the specific surface area and total pore volume of AC decrease from 2 137 m^2/g to 1 683 m^2/g, and 0.95 cm^3/g to 0.78 cm^3/g, respectively, if it is ball-milled for 8 h. The pore size distributions are similar in the range of 0.7 nm to 3.5 nm for different ball-milling time. There exists oxidation on the surface of AC during the ball-milling process and the ratios of O-C=O oxygen compositions increase whereas those of C-O, C=O decrease. The peeling strength of AC coated on current collector is almost inverse proportion with the particle size of AC as well as the resistance of EDLCs, and its capacitance decreases about 6%.