Study of hydrodynamic characteristics in tubular photobioreactors

In this work, the hydrodynamic characteristics in tubular photobioreactors with a series of helical static mixers built-in were numerically investigated using computational fluid dynamics (CFD). The influences of height and screw pitch of the helical static mixer and fluid inlet velocity on the cell trajectories, swirl numbers and energy consumption were examined. In order to verify the actual results for cultivation of microalgae, cultivation experiments of freshwater Chlorella sp. were carried out in photobioreactor with and without helical static mixer built-in at the same time. It was shown that with built-in helical static mixer, the mixing of fluid could be intensified, and the light/dark cycle could also be achieved which is of benefit for the growth of microalgae. The biomass productivity of Chlorella sp. in tubular photobioreactor with helical static mixer built-in was 37.26 % higher than that in the photobioreactor without helical static mixer.     

Non-hydroperoxy-type Peroxides as Autocatalysts of Lipid Autoxidation

In this paper, we present a simple and sensitive method for the assay of intracellular neutral lipids of the yeast, Lipomyces starkeyi. In this method, we used a tower-shaped mixer for cell disruption and applied a diagnostic triglycerides assay kit as a sensitive color developer. This mixer could treat 14 samples at one time. No effect of pH on cell disruption was detected within the pH range of 1.6 to 6.8. Twenty min was chosen for cell disruption and for homogeneous dispersion of intracellular neutral lipids. The analytical results agreed well with those obtained by three conventional methods. The new method is simpler and of good reliability.     

A continuous-flow capillary mixing method to monitor reactions on the microsecond time scale.

A continuous-flow capillary mixing apparatus, based on the original design of Regenfuss et al. (Regenfuss, P., R. M. Clegg, M. J. Fulwyler, F. J. Barrantes, and T. M. Jovin. 1985. Rev. Sci. Instrum. 56:283-290), has been developed with significant advances in mixer design, detection method and data analysis. To overcome the problems associated with the free-flowing jet used for observation in the original design (instability, optical artifacts due to scattering, poor definition of the geometry), the solution emerging from the capillary is injected directly into a flow-cell joined to the tip of the outer capillary via a ground-glass joint. The reaction kinetics are followed by measuring fluorescence versus distance downstream from the mixer, using an Hg(Xe) arc lamp for excitation and a digital camera with a UV-sensitized CCD detector for detection. Test reactions involving fluorescent dyes indicate that mixing is completed within 15 micros of its initiation and that the dead time of the measurement is 45 +/- 5 micros, which represents a >30-fold improvement in time resolution over conventional stopped-flow instruments. The high sensitivity and linearity of the CCD camera have been instrumental in obtaining artifact-free kinetic data over the time window from approximately 45 micros to a few milliseconds with signal-to-noise levels comparable to those of conventional methods. The scope of the method is discussed and illustrated with an example of a protein folding reaction.     

A low cost gas mixer

A low cost gas mixer for permanent gases and volatile agents is described. The apparatus uses the simple principle that if known volumes of gases are mixed at the same pressure and temperature, then a mixture of known composition results. The apparatus is shown to have an accuracy of better than ± 0.08% vol/vol for gases and volatile agents in two and three part mixtures. For permanent gases the mixer could be used accurately over the concentration range 1–99% vol/vol. For volatile agents the mixer was suitable for mixing concentrations in the low percentage ranges (< 5% vol/vol). Gas mixtures, including volatile agents, could be stored in the mixer for approximately 2 h without changes in concentration outside this limit of accuracy.     

A quick method to determine root biomass distribution in diameter classes

Describing root biomass distribution in diameter classes is a fundamental way to understand the relation between a plant and its surrounding soil. Current methods used for its measurement are not well adapted to large root systems. A new quick method is proposed for the measurement of diameter distribution in large root systems. It is based on the one used in pedology to assess soil granulometry. Roots are dried, cut in a mixer and placed on a sieve column; biomass distribution according to root diameter is assessed by weighting the biomass recovered in each sieve. The validity of the method was tested by comparing the sieving method results with those obtained on dried root systems with a digital image analysing system. A sensitivity analysis showed that the optimal rotation speed of the mixer was 2,000 rpm and the optimal sieving time was 22 min. The actual diameter distribution of artificial root mixtures of known root diameter distribution was closely correlated with the root biomass distribution measured by the sieving method (r ² = 0.87). Its application to four identical root systems resulted in values of biomass per diameter class with small standard errors. It is the first method allowing directly to measure biomass (and not length) distribution in diameter classes. It is quick, cheap and does not require root system sub-sampling; consequently, large root systems which were almost never studied can now be analysed. This method is thus adequate for repeated measurements of root diameter distribution in agronomical or ecological research.     

The possibility for improvement of ceramic membrane ultrafiltration of an enzyme solution

Ultrafiltration represents an attractive downstream processing technique for enzymes concentration and their primary purification. However, the process efficiency is often limited by protein fouling and shear-induced enzyme deactivation, resulting in permeate flux decline and loss of enzyme activity. The objective of this work was to investigate the possibility for improvement of ceramic membrane ultrafiltration of endo-pectinase solution. Experimental investigations were performed on a 5 nm ceramic membrane with the Kenics static mixer placed inside the membrane in order to improve the process performance. The use of the static mixer resulted in the flux improvement of about 45% at a volume concentration factor (VCF) of 3 leading to the reduction of operation time of 25% and the energy saving of about 40%. Although the rejection of endo-pectinase was higher than 96%, the extensive loss of the enzyme activity during operation indicated that the modification of the feed solution is essential for improved ultrafiltration performance. Addition of pectin to the original endo-pectinase solution led to a significant reduction of the enzyme deactivation: the enzyme activity yield was 90% at a VCF of 1.6 during operation with the static mixer.     

Laminar-flow fluid mixer for fast fluorescence kinetics studies

The ability to mix aqueous liquids on microsecond time scales, while consuming minimal amounts of sample and maintaining UV-visible optical access to the mixing region, is highly desirable for a range of biophysical studies of fast protein and nucleic acid interactions and folding. We have constructed a laminar coaxial jet mixer that allows the measurement of UV-excited fluorescence from nanoliter and microliter quantities of material, mixed at microsecond rates. The mixer injects a narrow cylindrical stream (radius a < 1 microm) of fluorescent sample into a larger flow of diluting buffer that moves through a capillary (100 microm i.d.) at a speed approximately 20 cm/s, under laminar flow conditions (Re approximately equal to 14). Construction from a fused silica capillary allows the laser excitation (at 266 nm) and detection (at 350 nm) of tryptophan fluorescence at reasonably low working concentrations, without interference from background fluorescence. Using this mixer we have measured sub-millisecond fluorescence quenching kinetics while consuming fluorescent sample at rates no greater than 6 nl/s. Consumption of the diluting buffer is also very modest (approximately 1-3 microl/s) in comparison with other rapid mixer designs.     

Numerical simulation and PEPT measurements of a 3D conical helical-blade mixer: a high potential solids mixer for solid-state fermentation

Helical-blade solids mixers have a large potential as bioreactors for solid-state fermentation (SSF). Fundamental knowledge of the flow and mixing behavior is required for robust operation of these mixers. In this study predictions of a discrete particle model were compared to experiments with colored wheat grain particles and positron emission particle tracking (PEPT) measurements. In the discrete particle model individual movements of particles were calculated from interaction forces. It was concluded that the predicted overall flow behavior matched well with the PEPT measurements. Differences between the model predictions and the experiments with wheat grains were found to be due to the assumption that substrate particles were spherical, which was in the model. Model simulations and experiments with spherical green peas confirmed this. The mixing in the helical-blade mixer could be attributed to (1) the transport of particles up and down in the interior of the mixer, and (2) dispersion or micro-mixing of particles in the top region of the mixer. It appeared that the mixing rate scaled linearly with the rotation rate of the blade, although the average particle velocity did not scale proportionally. It may be that the flow behavior changes as a function of the rotation rate (e.g., changing thickness of the top region); further study is required to confirm this. To increase the mixing performance of the mixer, a larger blade or a change in the shape of the mixer (larger top surface/volume ratio) is recommended.     

The effect of adding water into the mixer on pelleting efficiency and pellet quality in diets for finishing pigs without and with use of an expander

Two diets for finishing pigs were used to determine the effect of adding water into the mixer on processing characteristics and pellet quality. Diet 1 was based on barley, oats and soybean meal (barley-based diet) and Diet 2 was based on maize and soybean meal (maize-based diet). Both diets were produced without and with use of an expander. Adding up to 120 g water/kg into the mixer prior to steam conditioning and pelleting (steam conditioning) of the barley-based diet, improved pelleting efficiency, pellet durability index (PDI) and modified PDI (with five hexagonal nuts added into the tumbling box), by 15, 10 and 10%, respectively. When water was added into the mixer prior to expanding the barley-based diet, PDI increased from 93 to 95% and modified PDI from 91 to 94%. The maize-based diet was processed with motor load held constant to maximize feed mill output, and production rate held constant to minimize energy use. At a constant motor load and a constant production rate, adding up to 30 g water/kg into the maize-based diet prior to steam conditioning increased pelleting efficiency by 22 and 9%, PDI was improved from 84 to 89% and from 79 to 87%, respectively. Expander conditioning of the maize-based diet increased PDI from 92% at none to 94% at 30 g/kg water addition. Correspondingly, modified PDI was increased from 89 to 93% at none and 30 g water/kg into the mixer prior to expander conditioning. The overall conclusion from the experiment was that adding water into the mixer before steam conditioning improved pelleting efficiency and pellet quality in barley- and maize-based diet for finishing pigs.     

Molecular dynamics simulations of aqueous pullulan oligomers

Molecular dynamics (MD) simulations have been used to model small-angle X-ray scattering (SAXS) data on aqueous solutions of four oligomeric segments of the glucan pullulan: the trimer G(3) (comprising one polymer repeating unit), the hexamer (G(3))(2), the nonamer (G(3))(3), and the dodecamer (G(3))(4). The AMBER force field was used in conjunction with the GB/SA continuum solvation model to calculate both the mean global dimensions of the oligomers from the limiting small angle scattering behavior and the shorter range structural information implicit in the Debye scattering function at larger scattering angles. This same force field and solvation treatment were employed earlier by Liu et al. (Macromolecules 1999, 32, 8611-8620) with apparent success in a rotational isomeric state (RIS) treatment of the same experimental data. The present work discloses that, despite numerical success in modeling the SAXS data, the RIS treatment, which includes only the interactions within dimeric segments of the polymer chain, fails to account accurately for excluded volume effects at the range of 3-12 sugar residues in the polymer backbone. It is suggested that MD simulations using continuum solvation models can be used to circumvent errors inherent in the computationally efficient RIS treatments of polymer nano- and picosecond dynamics while at the same time avoiding the heavy computational requirements of all-atom methods.     

Evaluation of portable haemoglobinometer in general practice.

The HemoCue system for estimating haemoglobin was evaluated within urban general practice. It gave excellent results when used within a laboratory environment (on 103 paired samples) but disappointing ones when evaluated by practice nurses within general practice (on 235 paired samples). The most likely source of error was inadequate mixing of the blood specimens before sampling, which might be obviated by using a rotating mixer. It is emphasised that equipment intended for use in general practice should be evaluated under normal working conditions envisaged.     

Convex Grooves in Staggered Herringbone Mixer Improve Mixing Efficiency of Laminar Flow in Microchannel

The liquid streams in a microchannel are hardly mixed to form laminar flow, and the mixing issue is well described by a low Reynolds number scheme. The staggered herringbone mixer (SHM) using repeated patterns of grooves in the microchannel have been proved to be an efficient passive micro-mixer. However, only a negative pattern of the staggered herringbone mixer has been used so far after it was first suggested, to the best of our knowledge. In this study, the mixing efficiencies from negative and positive staggered herringbone mixer patterns as well as from opposite flow directions were tested to investigate the effect of the micro-structure geometry on the surrounding laminar flow. The positive herringbone pattern showed better mixing efficiency than the conventionally used negative pattern. Also, generally used forward flow gives better mixing efficiency than reverse flow. The mixing was completed after two cycles of staggered herringbone mixer with both forward and reverse flow in a positive pattern. The traditional negative pattern showed complete mixing after four and five cycles in forward and reverse flow direction, respectively. The mixing effect in all geometries was numerically simulated, and the results confirmed more efficient mixing in the positive pattern than the negative. The results can further enable the design of a more efficient microfluidic mixer, as well as in depth understanding of the phenomena of positive and negative patterns existing in nature with regards to the surrounding fluids.     

The effect of lintnerisation on wheat and potato starch granules

Wheat and potato starches were hydrolysed with 2·2 n hydrochloric acid at 35°C for a period of time up to 15 days. The residues (lintnerised starches) were washed and freeze dried, and studied by differential scanning calorimetry (DSC), wide-angle X-ray scattering (WAXS), small-angle light scattering (SALS), small-angle neutron scattering (SANS) and small-angle X-ray scattering (SAXS). These techniques showed that profound changes took place in the first day of hydrolysis (during which time the extent of hydrolysis was 7·7% for potato starch and 12·5% for wheat starch). In particular, the gelatinisation enthalpy (ΔH) decreased, the X-ray crystallinity increased and the SANS and SAXS peaks (indicative of a regular spacing between crystalline and amorphous regions) virtually disappeared. The reduction in ΔH is surprising and is discussed at length. It was also shown that freeze drying results in a considerable lowering of the gelatinisation temperature of potato starch (and also of ΔH) while that of wheat starch is only slightly affected.     

Separation and characterisation of light scattering transients from rod outer segments of vertebrate photoreceptors: design and performance of a Multi Angle Flash Photolysis Apparatus (MAFPA)

A device was built for the simple computer-controlled routine determination of the angular dependence of light scattering transients obtained from biological material. It was called Multi Angle Flash Photolysis Apparatus (MAFPA). The MAFPA allows the simultaneous registration of rapid, light-induced light scattering transients at eight scattering angles between 0 degree and 28 degrees. In typical applications changes in scattered light intensity as small as delta I/I = 4 X 10(-5) can be resolved at scattering angles less than 24 degrees, while at 28 degrees the resolution drops to delta I/I = 2 X 10(-4). The time resolution is 32 microseconds. The MAFPA was designed for high accuracy, ease of use and ruggedness. It is made from relatively inexpensive parts and can be copied fairly easily by a good machine/electronics shop. In this communication we describe the design of the MAFPA and how it was used for the characterisation of four structurally distinct light-induced light scattering signals from photoreceptor rod outer segments. These signals are known as P (or binding) signal, G- (or dissociation) signal, N (or rhodopsin) signal and as the ATP-dependent signal AL. The signals have been separated by means of their different angular dependence, their different saturation behavior and nucleotide requirement. A great number of detailed studies will have to be carried out before one can fully understand the physical and biochemical origin of these signals. At this point, however, it can be stated that the so-called 'dissociation signal', showing an angular dependence indicative of a change in refractive index or scattering mass, is not merely an inversion of the preceding 'binding signal', the latter clearly reflecting a gross structural change, i.e. a shrinkage of the disks. Moreover, there are conditions where P signals are observed to persist even after the completion of the subsequent dissociation signals. The two remaining signals N and AL show a pronounced angular dependence which is not easily interpreted. The fact that both exhibit a maximal amplitude at relatively small angles seems to indicate the participation of rather large structural domains.     

Temperature control in a continuously mixed bioreactor for solid-state fermentation

A continuously mixed, aseptic paddle mixer was used successfully for solid-state fermentation (SSF) with Aspergillus oryzae on whole wheat kernels. Continuous mixing improved temperature control and prevented inhomogeneities in the bed. Respiration rates found in this system were comparable to those in small, isothermal, unmixed beds, which showed that continuous mixing did not cause serious damage to the fungus or the wheat kernels. Continuous mixing improves heat transport to the bioreactor wall, which reduces the need for evaporative cooling and thus may help to prevent the desiccation problems that hamper large-scale SSF. However, scale-up calculations for the paddle mixer indicated that wall cooling becomes insufficient at the 2-m(3) scale for a rapidly growing fungus like Aspergillus oryzae. Consequently, evaporative cooling will remain important in large-scale mixed systems. Experiments showed that water addition will be necessary when evaporative cooling is applied in order to maintain a sufficiently high water activity of the solid substrate. Mixing is necessary to ensure homogeneous water addition in SSF. Automated process control might be achieved using the enthalpy balance. The enthalpy balance for the case of evaporative cooling in the paddle mixer was validated. This work shows that continuous mixing provides promising possibilities for simultaneous control of temperature and moisture content in solid-state fermentation on a large scale.     

Considerations of coherent scattering and electron binding in incoherent scattering, in computation of dose deposition in tissue from low-energy photon beams

Two different sets of Monte Carlo computations were carried out for the study of dose penetration of monoenergetic, low-energy (10 to 100 keV) photon beams incident on slabs of tissue. One program took into account coherent scattering and considered electron binding when finding the angle of scattering during incoherent scattering; the other simpler program, customarily used at higher energies, largely ignored these effects. For calculations at the source photon energy of 100 keV, it was found that there was negligible difference in dose distribution in the slab between the more and less complex type of calculations. The same thing was found to be true for the 30 and 10-keV source photon energies only for shallow penetration distances; and at deeper penetrations the simple approach tended to overestimate the dose appreciably. It is concluded that for penetration of low-energy photon beams into tissue, accurate calculational results cannot be assured with the neglect of coherent scattering effects and electron binding considerations in determining the scattering angles except for shallow depths of penetration.     

Simple and small-scale breakdown of yeast

A simple and small-scale method for the preparation of homogenate from yeast was developed. The principle of this new method involves the shaking of a small amount of yeast-cell suspension with glass beads on a tower-shaped mixer. When this method is used the cells in 1 ml of cell suspension are broken down and 14 samples can be simultaneously processed under controlled conditions. The degree of cell breakdown, the amount of soluble protein liberated from the cells, and the increase in each enzyme activity in prepared homogenate correlated to the shaking time: maximum values were obtained at 20 min. Enzyme activities in the homogenate were equivalent to or higher than that procured with former methods. This new method is applicable to various species of yeasts.     

Chaotic mixer improves microarray hybridization

Hybridization is an important aspect of microarray experimental design which influences array signal levels and the repeatability of data within an array and across different arrays. Current methods typically require 24h and use target inefficiently. In these studies, we compare hybridization signals obtained in conventional static hybridization, which depends on diffusional target delivery, with signals obtained in a dynamic hybridization chamber, which employs a fluid mixer based on chaotic advection theory to deliver targets across a conventional glass slide array. Microarrays were printed with a pattern of 102 identical probe spots containing a 65-mer oligonucleotide capture probe. Hybridization of a 725-bp fluorescently labeled target was used to measure average target hybridization levels, local signal-to-noise ratios, and array hybridization uniformity. Dynamic hybridization for 1h with 1 or 10ng of target DNA increased hybridization signal intensities approximately threefold over a 24-h static hybridization. Similarly, a 10- or 60-min dynamic hybridization of 10ng of target DNA increased hybridization signal intensities fourfold over a 24h static hybridization. In time course studies, static hybridization reached a maximum within 8 to 12h using either 1 or 10ng of target. In time course studies using the dynamic hybridization chamber, hybridization using 1ng of target increased to a maximum at 4h and that using 10ng of target did not vary over the time points tested. In comparison to static hybridization, dynamic hybridization reduced the signal-to-noise ratios threefold and reduced spot-to-spot variation twofold. Therefore, we conclude that dynamic hybridization based on a chaotic mixer design improves both the speed of hybridization and the maximum level of hybridization while increasing signal-to-noise ratios and reducing spot-to-spot variation.     

The effect of aeration and agitation on tetracycline biosynthesis

The results of the study on relation between tetracycline biosynthesis and the specific power input for agitation in pilot plant apparatus was studied. No correlation was observed between the levels of tetracycline biosynthesis and changes in the specific power input within a range of 0.6 to 2.3 kW/m3 at the expense of changes in the mixer diameter and the agitation rate, when the aeration rate was constant. It was shown that the aeration conditions were most significant for tetracycline biosynthesis. The study provided determination of the optimal aeration conditions for biosynthesis of tetracycline.     

Effects of hemolysis, hematocrit, RBC swelling, and flow rate on light scattering by blood in a 0.26 cm ID transparent tube

The intensity of light scattering by blood in a tube of diameter 0.26 cm was measured with an apparatus devised by us at different angles on an incident cross-sectional plane. Changes in angular distribution of light intensity associated with hemolysis, and changes in hematocrit (Ht), red blood cell (RBC) swelling, and flow rate were plotted on polar coordinates. The dyssymmetry index, defined as the ratio of the intensity of light at 45 degrees to that at 135 degrees, was used to characterize the shape of the diagrams of light scattering. The index decreased with Ht, flow rate, but increased with RBC swelling. It is concluded that light scattering by blood requires intactness of the RBC membrane. Even when the cell membrane is intact, light scattering is subject to changes with the flow rate of blood, presumably due to RBC aggregation.