Large-scale recovery of C-phycocyanin from Spirulina platensis using expanded bed adsorption chromatography

C-phycocyanin was purified on a large scale by a combination of expanded bed adsorption, anion-exchange chromatography and hydroxyapatite chromatography from inferior Spirulina platensis that cannot be used for human consumption. First, phycobiliproteins were extracted by a simple, scaleable method and then were recovered by Phenyl-Sepharose chromatography in an expanded bed column. The purity (the A(620)/A(280) ratio) of C-phycocyanin isolated with STREAMLINE (TM) Column was up to 2.87, and the yield was as high as 31 mg/g of dried S. platensis. After the first step, we used conventional anion-exchange chromatography for the purification steps, with a yield of 7.7 mg/g of dried S. platensis at a purity greater than 3.2 and with an A(620)/A(650) index higher than 5.0. The fractions from anion-exchange chromatography with a level of purity that did not conform to the above standard were subjected to hydroxyapatite chromatography, with a C-PC yield of 4.45 mg/g of dried S. platensis with a purity greater than 3.2. The protein from both purification methods showed one absolute absorption peak at 620 nm and a fluorescence maximum at 650 nm, which is consistent with the typical spectrum of C-phycocyanin. SDS-PAGE gave two bands corresponding to 21 and 18 kDa. In-gel digestion and LC-ESI-MS showed that the protein is C-phycocyanin. (c) 2006 Elsevier B.V. All rights reserved.

Purification of Antibodies Specific to a Dinucleotide Using the Hapten Bound to Deae Cellulose as an Affinity Column

The dinucleotide dpTpA held electrostatically on DEAE cellulose was used as an affinity column for the purification of dpTpA specific antibodies. Chromatography of the y-globulin fraction from dpTpA specific antisera on this column resulted in the retention of dpTpA specific antibodies which were later eluted along with the bound dpTpA using 1M NaC1. Dextran coated charcoal was the method of choice for the dissociation and removal of dpTpA bound to the antibodies. This method may extend itself to the purification of antibodies specific for other oligonucleotides.

Recuperação e purificação de quitosanases usando adsorção em leito expandido com streamline DEAE com modelagem e simulação usando redes neurais

Expanded Bed Adsorption (EBA) is an integrative process that combines concepts of chromatography and fluidization of solids. The many parameters involved and their synergistic effects complicate the optimization of the process. Fortunately, some mathematical tools have been developed in order to guide the investigation of the EBA system. In this work the application of experimental design, phenomenological modeling and artificial neural networks (ANN) in understanding chitosanases adsorption on ion exchange resin Streamline® DEAE have been investigated. The strain Paenibacillus ehimensis NRRL B-23118 was used for chitosanase production. EBA experiments were carried out using a column of 2.6 cm inner diameter with 30.0 cm in height that was coupled to a peristaltic pump. At the bottom of the column there was a distributor of glass beads having a height of 3.0 cm. Assays for residence time distribution (RTD) revelead a high degree of mixing, however, the Richardson-Zaki coefficients showed that the column was on the threshold of stability. Isotherm models fitted the adsorption equilibrium data in the presence of lyotropic salts. The results of experiment design indicated that the ionic strength and superficial velocity are important to the recovery and purity of chitosanases. The molecular mass of the two chitosanases were approximately 23 kDa and 52 kDa as estimated by SDS-PAGE. The phenomenological modeling was aimed to describe the operations in batch and column chromatography. The simulations were performed in Microsoft Visual Studio. The kinetic rate constant model set to kinetic curves efficiently under conditions of initial enzyme activity 0.232, 0.142 e 0.079 UA/mL. The simulated breakthrough curves showed some differences with experimental data, especially regarding the slope. Sensitivity tests of the model on the surface velocity, axial dispersion and initial concentration showed agreement with the literature. The neural network was constructed in MATLAB and Neural Network Toolbox. The cross-validation was used to improve the ability of generalization. The parameters of ANN were improved to obtain the settings 6-6 (enzyme activity) and 9-6 (total protein), as well as tansig transfer function and Levenberg-Marquardt training algorithm. The neural Carlos Eduardo de Araújo Padilha dezembro/2013 9 networks simulations, including all the steps of cycle, showed good agreement with experimental data, with a correlation coefficient of approximately 0.974. The effects of input variables on profiles of the stages of loading, washing and elution were consistent with the literature

Cyanobacteria detection in Guarapiranga Reservoir (São Paulo State, Brazil) using Landsat TM and ETM+ images

Algae bloom is one of the major consequences of the eutrophication of aquatic systems, including algae capable of producing toxic substances. Among these are several species of cyanobacteria, also known as blue-green algae, that have the capacity to adapt themselves to changes in the water column. Thus, the horizontal distribution of cyanobacteria harmful algae blooms (CHABs) is essential, not only to the environment, but also for public health. The use of remote sensing techniques for mapping CHABs has been explored by means of bio-optical modeling of phycocyanin (PC), a unique inland waters cyanobacteria pigment. However, due to the small number of sensors with a spectral band of the PC absorption feature, it is difficult to develop semi-analytical models. This study evaluated the use of an empirical model to identify CHABs using TM and ETM+ sensors aboard Landsat 5 and 7 satellites. Five images were acquired for applying the model. Besides the images, data was also collected in the Guarapiranga Reservoir, in São Paulo Metropolitan Region, regarding the cyanobacteria cell count (cells/mL), which was used as an indicator of CHABs biomass. When model values were analyzed excluding calibration factors for temperate lakes, they showed a medium correlation (R²=0.81, p=0.036), while when the factors were included the model showed a high correlation (R²=0.96, p=0.003) to the cyanobacteria cell count. The empirical model analyzed proved useful as an important tool for policy makers, since it provided information regarding the horizontal distribution of CHABs which could not be acquired from traditional monitoring techniques.

Hydrodynamics and mass transfer coefficient in activated sludge aerated stirred column reactor: Experimental analysis and modeling

The aerated stirred reactor (ASR) has been widely used in biochemical and wastewater treatment processes. The information describing how the activated sludge properties and operation conditions affect the hydrodynamics and mass transfer coefficient is missing in the literature. The aim of this study was to investigate the influence of flow regime, superficial gas velocity (U-G), power consumption unit (P/V-L), sludge loading, and apparent viscosity (pap) of activated sludge fluid on the mixing time (t(m)), gas hold-up (epsilon), and volumetric mass transfer coefficient (kLa) in an activated sludge aerated stirred column reactor (ASCR). The activated sludge fluid performed a non-Newtonian rheological behavior. The sludge loading significantly affected the fluid hydrodynamics and mass transfer. With an increase in the UG and P/V-L, the epsilon and k(L)a increased, and the t(m), decreased. The E, kLa, and tm,were influenced dramatically as the flow regime changed from homogeneous to heterogeneous patterns. The proposed mathematical models predicted the experimental results well under experimental conditions, indicating that the U-G, P/V-L, and mu(ap) had significant impact on the t(m) epsilon, and k(L)a. These models were able to give the tm, F, and kLa values with an error around +/- 8%, and always less than +/- 10%. (c) 2005 Wiley Periodicals, Inc.

Experimental Infection of Culex Annulirostris, Culex Gelidus, and Aedes Vigilax With a Yellow Fever/Japanese Encephalitis Virus Vaccine Chimera (Chimerivax TM-JE)

Australian mosquitoes from which Japanese encephalitis virus (JEV) has been recovered (Culex annulirostris, Culex gelidus, and Aedes vigilax) were assessed for their ability to be infected with the ChimeriVax-JE vaccine, with yellow fever vaccine virus 17D (YF 17D) from which the backbone of ChimeriVax-JE vaccine is derived and with JEV-Nakayama. None of the mosquitoes became infected after being fed orally with 6.1 log(10) plaque-forming units (PFU)/mL of ChimeriVax-JE vaccine, which is greater than the peak viremia in vaccinees (mean peak viremia = 4.8 PFU/mL, range = 0-30 PFU/mL of 0.9 days mean duration, range = 0-11 days). Some members of all three species of mosquito became infected when fed on JEV-Nakayama, but only Ae. vigilax was infected when fed on YF 17D. The results suggest that none of these three species of mosquito are likely to set up secondary cycles of transmission of ChimeriVax-JE in Australia after feeding on a viremic vaccinee.

Zeolite from alkali modified kaolin increases NH4+ retention by sandy soil : column experiments

Sandy soils have low water and nutrient retention capabilities so that zeolite soil amendments are used for high value land uses including turf and horticulture to reduce leaching losses of NH4+ fertilisers. MesoLite is a zeolitic material made by caustic treatment of kaolin at 80-95oC. It has a moderately low surface area (9-12m2/g) and very high cation exchange capacity (494 cmol(+)/kg). Laboratory column experiments showed that an addition of 0.4% MesoLite to a sandy soil greatly (90%) reduced leaching of added NH4+ compared to an unamended soil and MesoLite is 11 times more efficient in retaining NH4+ than natural zeolite. Furthermore, NH4+-MesoLite slowly releases NH4+ to soil solution and is likely to be an effective slow release fertiliser.

Bolted beam-column moment connections between cold-formed steel members

ABSTRACT Twelve beam-to-column connections between cold-formed steel sections consisting of three beam depths and four connection types were tested in isolation to investigate their behavior based on strength, stiffness and ductility. Resulting moment-rotation curves indicate that the tested connections are efficient moment connections where moment capacities ranged from about 65% to 100% of the connected beam capac-ity. With a moment capacity of greater than 80% of connected beam member capacity, some of the connec-tions can be regarded as full strength connections. Connections also possessed sufficient ductility with rota-tions of 20 mRad at failure although some connections were too ductile with rotations in excess of 30 mRad. Generally, most of the connections possess the strength and ductility to be considered as partial strength con-nections. The ultimate failures of almost all of the connections were due to local buckling of the compression web and flange elements of the beam closest to the connection.

The NetWorkPlace(TM) Phenomenon: any time and any place - reality or a seductive myth?

A new spatial logic encompassing redefined concepts of time and place, space and distance, requires a comprehensive shift in the approach to designing workplace environments for today’s adaptive, collaborative organizations operating in a dynamic business world. Together with substantial economic and cultural shifts and an increased emphasis on lifestyle considerations, the advances in information technology have prompted a radical re-ordering of organizational relationships and the associated structures, processes, and places of doing business. Within the duality of space and an augmentation of the traditional notions of place, organizational and institutional structures pose new challenges for the design professions. The literature reveals that there has always been a mono-organizational focus in relation to workplace design strategies and the burgeoning trend towards inter-organizational collaboration, enabled the identification of a gap in the knowledge relative to workplace design. The NetWorkPlaceTM© constitutes a multi-dimensional concept having the capacity to deal with the fluidity and ambiguity characteristic of the network context, as both a topic of research and the way of going about it.

Blast response and safety evaluation of a composite column for use as key element in structural systems

This paper presents the blast response, damage mechanism and evaluation of residual load capacity of a concrete–steel composite (CSC) column using dynamic computer simulation techniques. This study is an integral part of a comprehensive research program which investigated the vulnerability of structural framing systems to catastrophic and progressive collapse under blast loading and is intended to provide design information on blast mitigation and safety evaluation of load bearing vulnerable columns that are key elements in a building. The performance of the CSC column is compared with that of a reinforced concrete (RC) column with the same dimensions and steel ratio. Results demonstrate the superior performance of the CSC column, compared to the RC column in terms of residual load carrying capacity, and its potential for use as a key element in structural systems. The procedure and results presented herein can be used in the design and safety evaluation of key elements of multi-storey buildings for mitigating the impact of blast loads.

Comparison of three generations of ActiGraph (TM) activity monitors in children and adolescents

In this study, we evaluated agreement among three generations of ActiGraph (TM) accelerometers in children and adolescents. Twenty-nine participants (mean age = 14.2 +/- 3.0 years) completed two laboratory-based activity sessions, each lasting 60 min. During each session, participants concurrently wore three different models of the ActiGraph (TM) accelerometers (GT1M, GT3X, GT3X+). Agreement among the three models for vertical axis counts, vector magnitude counts, and time spent in moderate-to-vigorous physical exercise (MVPA) was evaluated by calculating intraclass correlation coefficients and Bland-Altman plots. The intraclass correlation coefficient for total vertical axis counts, total vector magnitude counts, and estimated MVPA was 0.994 (95% CI = 0.989-0.996), 0.981 (95% CI = 0.969-0.989), and 0.996 (95% CI = 0.989-0.998), respectively. Inter-monitor differences for total vertical axis and vector magnitude counts ranged from 0.3% to 1.5%, while inter-monitor differences for estimated MVPA were equal to or close to zero. On the basis of these findings, we conclude that there is strong agreement between the GT1M, GT3X, and GT3X+ activity monitors, thus making it acceptable for researchers and practitioners to use different ActiGraph (TM) models within a given study.