Relation between cell disruption conditions, cell debris particle size, and inclusion body release

The efficiency of physical separation of inclusion bodies from cell debris is related to cell debris size and inclusion body release and both factors should be taken into account when designing a process. In this work, cell disruption by enzymatic treatment with lysozyme and cellulase, by homogenization, and by homogenization with ammonia pretreatment is discussed. These disruption methods are compared on the basis of inclusion body release, operating costs, and cell debris particle size. The latter was measured with cumulative sedimentation analysis in combination with membrane-associated protein quantification by SDS-PAGE and a spectrophotometric pepticloglycan quantification method. Comparison of the results obtained with these two cell debris quantification methods shows that enzymatic treatment yields cell debris particles with varying chemical composition, while this is not the case with the other disruption methods that were investigated. Furthermore, the experiments show that ammonia pretreatment with homogenization increases inclusion body release compared to homogenization without pretreatment and that this pretreatment may be used to control the cell debris size to some extent. The enzymatic disruption process gives a higher product release than homogenization with or without ammonia pretreatment at lower operating costs, but it also yields a much smaller cell debris size than the other disruption process. This is unfavorable for centrifugal inclusion body purification in this case, where cell debris is the component going to the sediment and the inclusion body is the floating component. Nevertheless, calculations show that centrifugal separation of inclusion bodies from the enzymatically treated cells gives a high inclusion body yield and purity. (C) 2004 Wiley Periodicals, Inc.

Focused beam reflectance technique for in situ particle sizing in wastewater treatment settling tanks

Due to the complexities involved with measuring activated sludge floc size distributions, this parameter has largely been ignored by wastewater researchers and practitioners. One of the major reasons has been that instruments able to measure particle size distributions were complex, expensive and only provided off-line measurements. The Focused Beam Reflectance Method (FBRM) is one of the rare techniques able to measure the particle size distribution in situ. This paper introduces the technique for monitoring wastewater treatment systems and compares its performance with other sizing techniques. The issue of the optimal focal point is discussed, and similar conclusions as found in the literature for other particulate systems are drawn. The study also demonstrates the capabilities of the FBRM in evaluating the performance of settling tanks. Interestingly, the floc size distributions did not vary with position inside the settling tank flocculator. This was an unexpected finding, and seriously questioned the need for a flocculator in the settling tank. It is conjectured that the invariable size distributions were caused by the unique combination of high solids concentration, low shear and zeolite dosing. (C) 2004 Society of Chemical Industry.

Particle bed charge decay behaviour under high tension roll separation

A new theory of particle discharge in high tension roll (HTR) separation is presented. The discharge dynamics of an isolated charged particle resting on a conducting surface are studied first. The analysis is extended to particle discharge in a homogenous particle bed. Finally, the paper looks at the more realistic scenario of particle discharge in a non-homogenous particle bed. The consequences of the resulting theory on HTR separation are discussed. Predictions from the new theory are tested against experimental HTR separations at the pilot scale. (c) 2005 Elsevier Ltd. All rights reserved.

Aquatic passive sampling of herbicides on naked particle loaded membranes: Accelerated measurement and empirical estimation of kinetic parameters

Water-sampler equilibrium partitioning coefficients and aqueous boundary layer mass transfer coefficients for atrazine, diuron, hexazionone and fluometuron onto C18 and SDB-RPS Empore disk-based aquatic passive samplers have been determined experimentally under a laminar flow regime (Re = 5400). The method involved accelerating the time to equilibrium of the samplers by exposing them to three water concentrations, decreasing stepwise to 50% and then 25% of the original concentration. Assuming first-order Fickian kinetics across a rate-limiting aqueous boundary layer, both parameters are determined computationally by unconstrained nonlinear optimization. In addition, a method of estimation of mass transfer coefficients-therefore sampling rates-using the dimensionless Sherwood correlation developed for laminar flow over a flat plate is applied. For each of the herbicides, this correlation is validated to within 40% of the experimental data. The study demonstrates that for trace concentrations (sub 0.1 mu g/L) and these flow conditions, a naked Empore disk performs well as an integrative sampler over short deployments (up to 7 days) for the range of polar herbicides investigated. The SDB-RPS disk allows a longer integrative period than the C18 disk due to its higher sorbent mass and/or its more polar sorbent chemistry. This work also suggests that for certain passive sampler designs, empirical estimation of sampling rates may be possible using correlations that have been available in the chemical engineering literature for some time.

Segmented polyurethane nanocomposites: Impact of controlled particle size nanofillers on the morphological response to uniaxial deformation

A series of TPU nanocomposites were prepared by incorporating organically modified layered silicates with controlled particle size. To our knowledge, this is the first study into the effects of layered silicate diameter in polymer nanocomposites utilizing the same mineral for each size fraction. The tensile properties of these materials were found to be highly dependent upon the size of the layered silicates. A decrease in disk diameter was associated with a sharp upturn in the stress-strain curve and a pronounced increase in tensile strength. Results from SAXS/SANS experiments showed that the layered silicates did not affect the bulk TPU microphase structure and the morphological response of the host TPU to deformation or promote/hinder strain-induced soft segment crystallization. The improved tensile properties of the nanocomposites containing the smaller nanofillers resulted from the layered silicates aligning in the direction of strain and interacting with the TPU sequences via secondary bonding. This phenomenon contributes predominantly above 400% strain once the microdomain architecture has largely been disassembled. Large tactoids that are unable to align in the strain direction lead to concentrated tensile stresses between the polymer and filler, instead of desirable shear stresses, resulting in void formation and reduced tensile properties. In severe cases, such as that observed for the composite containing the largest silicate, these voids manifest visually as stress whitening.

Exploratory modelling of grinding pressure within a compressed particle bed

With increasing industry interest in high pressure roll grinding (HPGR) technology, there is a strong incentive for improved understanding of the nature of grinding pressure that exists in the interior of a compressed particle bed. This corresponds to the crushing region of the HPGR. The relationship between applied pressure (stress) to the particle bed and induced pressure (stress) within particles and at contact points between particles is of particular interest. A detailed parametric investigation is beyond the scope of this exploratory paper. However, this exploratory investigation does suggest some interesting behaviour. The compressed particle bed within an 80 turn diameter piston has been modelled using Particle Flow Code for three dimensions. PFC3D is a discrete element code. The total number of simulated particles was 1225 and 2450 for two beds of different thickness. Particle diameters were uniformly distributed between 4 and 4.5 mm. The results of the simulations show that stress intensity within the simulated particle beds and within the observed particles increased with increase of the applied stress. The intensity of the average vertical stress in the selected particles tended to be comparable with the intensity of the pressure applied to the surface of particle bed and was only occasionally higher. However, the stress at contact points between particles could be several times higher. In a real crusher, such high stress amplification at contacts will quickly decrease due to local crushing and a resultant increase the size of the contact area. Therefore, its significance is likely to be relatively small in an industrial context. The modelling results also suggest that failure within the particle bed will progress from the crushing surface towards the depth of the bed. (c) 2006 Elsevier Ltd. All rights reserved.

Conditional quantum-state engineering using ancillary squeezed-vacuum states

We investigate an optical scheme to conditionally engineer quantum states using a beam splitter, homodyne detection, and a squeezed vacuum as an ancillar state. This scheme is efficient in producing non-Gaussian quantum states such as squeezed single photons and superpositions of coherent states (SCSs). We show that a SCS with well defined parity and high fidelity can be generated from a Fock state of n

Engineering of needle-free physical methods to target epidermal cells for DNA vaccination

A challenge in epidermal DNA vaccination is the efficient and targeted delivery of polynucleotides to immunologically sensitive Langerhans cells. This paper investigates this particular challenge for physical delivery approaches. The skin immunology and material properties are examined in the context of the physical cell targeting requirements of the viable epidermis. Selected current physical cell targeting technologies engineered to meet these needs are examined: needle and syringe; diffusion patches; liquid jet injectors; microneedle arrays/patches; and biolistic particle injection. The operating methods and relative performance of these approaches are discussed, with a comment on potential future developments and technologies. (c) 2005 Elsevier Ltd. All rights reserved.

Particle number emissions and source signatures of an industrial facility

The work presented was conducted within the scope of a larger study investigating impacts of the Stuart Oil Shale project, a facility operating to the north of the industrial city of Gladstone, Australia. The aims of the investigations were threefold: (a) the identification of the plant signatures in terms of particle size distributions in the submicrometer range (13-830 nm) through stack measurements, (b) exploring the applicability of these signatures in tracing the source contributions at locations of interest, at a distance from the plant, and (c) assessing the contribution of the plant to the total particle number concentration at locations of interest. The stack measurements conducted for three different conditions of plant operation showed that the particle size distributions were bimodal with average modal count median diameters (CMDs) of 24 (SD 4) and 52 (SD 9) nm. The average of all the particle size distributions recorded within the plant sector at a site located 4.5 km from the plant, over the sampling period when the plant was operating, also showed a bimodal distribution. The modal CMDs in this case were 27 and 50 nm, similar to those at the stack. This bimodal size distribution is distinct from the size distribution of the most common ambient anthropogenic emission source, which is vehicle emissions, and can be considered as a signature of this source. The average contribution of the plant (for plant sector winds) was estimated to be (10.0 +/- 3.8) x 10(2) particles cm(-3) and constituted approximately a 50% increase over the local particle ambient concentration for plant sector winds. This increase in particle number concentration compared to the local background concentration, while high compared to the clean environment concentration, is not significant when compared to concentrations generally encountered in the urban environment of Brisbane.