Electrical Sizing of Particles in Suspensions: III. Rigid Spheroids and Red Blood Cells

The processes involved during the passage of a suspended particle through a small cylindrical orifice across which exists an electric field are investigated experimentally for an approximate prolate spheroid in the form of two tangent, rigid spheres (ragweed pollen particles) and for fresh, human red blood cells. Oscillograms of current pulses produced by both types of particles are presented and discussed in terms of particle shape and orientation and the effects of the hydrodynamic field. It is concluded that all the particles enter the orifice with their major axes aligned parallel to the orifice axis (electric field), but that during their passage some are rotated by the hydrodynamic field. Cells with their equatorial plane perpendicular to a radius of the orifice change their orientation with respect to the electric field as they are rotated, the others do not; only in the former case is there any deformation. It is shown that the bimodal or skewed size distributions can be explained on this basis, and that size (shape factor × volume) is actually a normally distributed variable (P > 95%). The average size of samples from 10 healthy adults was found to be 102.7 μ³ with a coefficient of variation of 1.8%. For a volume of 87 μ³, this corresponds to a shape factor of 1.18, an axial ratio (assuming a perfect oblate spheroid) of 0.26, and an equivalent major axis of 8.6 μ. The effect of high electric fields on red cell size distributions is mentioned.     

Basic principles of electrical sizing of cells and particles and their realization in the new instrument "Metricell".

To understand the basic events during the passage of particles through the Coulter orifice, three experiments were performed. (1) The denpendence of the volume results on the particle path has been shown by ink-colored particle beams. (2) The deformation and alignment of cells during their passage through the orifice have been photographed by a nano-second photographing technique. (3) The absolute volume evaluation of particles has been studied with model particles in enlarged model orifices of different lengths. A new compact sizing instrument, "Metricell," equipped with a particle-independent electrical calibrating system, is described.     

Specific problems using electronic particle counters

Some specific problems of electronic particle counters (Coulter Counter, Elzone) are discussed. Conductivity of the medium does not influence the size response of the instrument, but might change the size of the particles through osmotic stress. An important problem of electronic particle counters is count loss by coincidence of passage of particles through the orifice and caused by dead time of the instruments. Several correction equations were tried out. It turned out that the count losses were much higher than the ones predicted by the manufacturers. In instruments equipped with multi-channel size analysers dead time count loss is much more important than the count loss by coincidence. A maximal counting frequency of 200 counts/second is recommended for accurate work. The width of the size distribution of particles depended on the diameter of the aperture tube.     

Supercritical assisted atomization: A novel technology for microparticles preparation of an asthma-controlling drug

The objective of this study was to produce microparticles of a new asthma-controlling drug by supercritical assisted atomization (SAA), proposed as an alternative to conventional jet-milling process. SAA is based on the solubilization of supercritical carbon dioxide in a liquid solution containing the drug; the ternary mixture is then sprayed through a nozzle, and microparticles are formed as a consequence of the enhanced atomization. SAA process parameters studied were precipitator temperature, nozzle diameter, and drug concentration in the liquid solution. Their influence was evaluated on morphology and size of precipitated particles. Spherical particles with mean particle size ranging from 1 to 3 μm of the new anti-asthma drug were produced by SAA. The mass median aerodynamic diameter (MMAD) of the SAA micronized particles and of the conventional jet-milled drug was used to compare, the results obtainable using the 2 techniques. Particularly, MMADs from 1.6 to 4.0 μm were obtained by SAA at the optimum operating conditions and by varying the concentration of the solution injected. MMAD of 6.0 μm was calculated for the jet-milled drug. SAA samples also exhibited narrower particle size distribution (PSD). A good control of particle size and distribution together with no drug degradation was obtained by SAA process. Published: October 22, 2005     

Formulation of a dry powder influenza vaccine for nasal delivery

The purpose of this research was to prepare a dry powder vaccine formulation containing whole inactivated influenza virus (VIIV) and a mucoadhesive compound suitable for nasal delivery. Powders containing WIIV and either lactose or trehalose were produced by lyophilization. A micro-ball mill was used to reduce the lyophilized cake to sizes suitable for nasal delivery. Chitosan flakes were reduced in size using a cryo-milling technique. Milled powders were sieved between 45 and 125 μm aggregate sizes and characterized for particle size and distribution, morphology, and flow properties. Powders were blended in the micro-ball mill without the ball. Lyophilization followed by milling produced irregularly shaped, polydisperse particles with a median primary particle diameter of ≈21 μm and a yield of ≈37% of particles in the 45 to 125 μm particle size range. Flow properties of lactose and trehalose powders after lyophilization followed by milling and sieving were similar. Cryo-milling produced a small yield of particles in the desired size range (<10%). Lyophilization followed by milling and sieving produced particles suitable for nasal delivery with different physicochemical properties as a function of processing conditions and components of the formulation. Further optimization of particle size and morphology is required for these powders to be suitable for clinical evaluation. Published: March 10, 2006     

Size, composition and distribution of particles related to wind induced resuspension in a shallow tropical lagoon

The size, composition and distribution of particles in the watercolumn were surveyed in a shallow area (1 m depth) of a tropicallagoon (Cte d'Ivoire) during a sequence of wind-induced resuspension.Water samples were collected hourly near the surface duringone tidal cycle. Three characteristic periods were distinguished:a calm period with low wind speed (average 1.2 m s^–1 awindy period with wind speed >3 m^–1 s (range between4 and 6 m s^–1) inducing sediment resuspension and a relaxationperiod during the decrease of wind velocity. From the analysisof several parameters (particle size and volume, bacteria. pico-and nanophytoplankton, ciliates and detritus), sediment resuspensioncaused a regular injection of particles from the bed. The finestparticles (1.5–6 µm: chlorophytes such as Chiorellaspp., picocyanobacteria such as Synechococcus) were the firstto be affected by wind-induced turbulence, whereas large particles(6–12 µm: diatoms. cyanobacteria such as Lyngbiaspp.) were dispersed into the water column at the highest windspeed. The fate of the different seston components differedaccording to their size. Therefore, wind-induced resuspensioncould greatly influence the food web organization through thequantity, quality and size of edible particles available ata given time.     

Phospholipid-stabilized nanoparticles of cyclosporine a by rapid expansion from supercritical to aqueous solution

The purpose of this research was to form stable suspensions of submicron particles of cyclosporine A, a water-insoluble drug, by rapid expansion from supercritical to aqueous solution (RESAS). A solution of cyclosporine A in CO₂ was expanded into an aqueous solution containing phospholipid vesicles mixed with nonionic surfactants to provide stabilization against particle growth resulting from collisions in the expanding jet. The products were evaluated by measuring drug loading with high performance liquid chromatography (HPLC), particle sizing by dynamic light scattering (DLS), and particle morphology by transmission electron microscopy (TEM) and x-ray diffraction. The ability of the surfactant molecules to orient at the surface of the particles and provide steric stabilization could be manipulated by changing process variables including temperature and suspension concentration. Suspensions with high payloads (up to 54 mg/mL) could be achieved with a mean diameter of 500 nm and particle size distribution ranging from 40 to 920 nm. This size range is several hundred nanometers smaller than that produced by RESAS for particles stabilized by Tween 80 alone. The high drug payloads (≈10 times greater than the equilibrium solubility), the small particle sizes, and the long-term stability make this process attractive for development.     

Electrical Sizing of Particles in Suspensions: I. Theory

The processes involved during the passage of a suspended particle through a small cylindrical orifice across which exists an electric field are considered in detail. Expressions are derived for the resulting change in current in terms of the ratios of particle to orifice volume and particle to suspending fluid resistivity, and particle shape. Graphs are presented of the electric field and of the fluid velocity as functions of position within the orifice, and of the shape factor of spheroids as a function of their axial ratio and orientation in the electric field. The effects of the electric and hydrodynamic fields on the orientation of nonspherical particles and on the deformation of nonrigid spheres is treated, and the migration of particles towards the orifice axis is discussed. Oscillograms of current pulses produced by rigid, nonconducting spheres in various orifices are shown and compared with the theoretical predictions.     

Grazing Rates in Euplotes mutabilis: Relationship between Particle Size and Concentration

Abstract Grazing behavior of both individual cells and populations of the marine hypotrich Euplotes mutabilis, a largely benthic ciliate that feeds on suspended particles, was studied using fluorescent latex microspheres. Microspheres of sizes 0.57-, 1.90-, 3.06-, 5.66-, and 10.0-μm diam were offered at concentrations from 10² to 10⁶ ml⁻¹. Their uptake in a ten-min period was determined. Food particles within such ranges of size and concentration are found under natural conditions. The ciliates ingested particles of all sizes offered. Uptake rates at all concentrations were dependent upon particle size, with 1.90- and 3.06-μm diam microspheres having the highest uptake rate in all cases. For all sizes, there was an increase in the percentage of feeding cells and in the uptake rate (the number of particles ingested cell⁻¹ h⁻¹), with increasing particle concentration. When uptake was expressed as the volume ingested, maximum values were obtained for 5.85-μm diam microspheres at a concentration of 10⁶ ml⁻¹. By taking a small number of large particles, present at a low concentration in the medium, a ciliate can ingest a greater biovolume than by taking a high number of small particles which are abundant in the medium. These results demonstrate that some ciliates can graze particles of a wide range of sizes. Also, its nutrition can be affected by changing ambient concentrations of different prey, both through effects on the proportion of feeding cells and through the relative food content of the particles. The data can also add to the understanding of food niche partitioning between species. At low particle concentrations, particularly, it is important to consider the behavior of individual ciliates as well as of the whole population. Received: 11 February 1997; Accepted: 21 October 1997     

Particle size effects in bioleaching of pyrite by acidophilic thermophile Sulfolobus metallicus (BC)

The effect of mineral particle size on the bioleaching of pyrite by the acidophilic thermophile Sulfolobus metallicus was investigated in a batch bioreactor. Decreasing the particle size from a mean diameter of 202 micron (size fraction: 150–180 micron) to a mean diameter of 42.5 micron (size fraction: 25–45 micron) enhanced the bioleaching rate from 0.05 kg m⁻³ h⁻¹ to 0.098 kg m⁻³ h⁻¹. The particle size distribution of the mineral in this range did not influence the morphology and growth kinetics of the cells. The values of specific growth rate (μ) and yield factor (Y) were 0.018–0.025 h⁻¹ and 0.67 × 10¹¹–1.45 × 10¹¹ cells (g iron)⁻¹, respectively. Decreasing the particle size of the mineral to a mean diameter of 6.40 micron (size fraction <25 micron) adversely influenced the activity of the cells. The presence of fine particles apparently damaged the structure of the cells, resulting in their inability to oxidise pyrite. Received: 11 December 1998 / Accepted: 9 April 1999     

A method for studying insoluble immune complexes

Two variants of a method for determining the average composition of insoluble immune complex particles (IICP) are described. The first variant is based on measuring the specific turbidity (the turbidity per unit mass concentration of the dispersed substance) and the average size of IICP determined from dynamic light scattering (DLS). In the second variant, the slope of the logarithmic turbidity spectrum (wavelength exponent) is used instead of DLS particle size. Both variants allow the average biopolymer volume fraction to be determined in terms of the average refractive index of IICP. The method is exemplified by two experimental antigen+antibody systems: (i) lipopolysaccharide-protein complex (LPPC) of Azospirillum brasilense Sp245+rabbit anti-LPPC; and (ii) human IgG (hIgG)+sheep anti-hIgG. We have found that IICP can be modeled by incompact porous particles that contain about 30% of biopolymer substance and 70% of buffer.     

Measurement of resuspended aerosol in the Chernobyl area

During anthropogenic activities, such as agricultural soil management and traffic on unpaved roads, size distribution measurements were performed of atmospheric particulate radionuclides at a site in the Chernobyl 30-km exclusion zone. Analysis of cascade impactor measurements showed an increase of the total atmospheric radioactivity. In the cases of harrowing by a tractor and traffic on unpaved roads, a common shape of the size distribution was found with two maxima, the first in the 2–4 μm range, the second in the 12–20 μm range. The size distributions were compared to measurements during wind-driven resuspension. Particle number concentration measurements with an Aerodynamic Particle Sizer showed a dynamic dependence of the particle concentration in different size ranges on anthropogenic action. The increase of the mean concentration was for the large particles more than one order of magnitude higher than for fine particles during anthropogenic enhanced resuspension. From the measurement of the mass concentration, the radioactive loading could be estimated. An enrichment of radionuclides on resuspended particles (compared to soil particles) was found, with the highest enrichment for large particles. Micrometeorological considerations showed that large particles may frequently be subject to medium range transport. The dry deposition velocity was measured; the mean value of 0.026 m s^–1±0.016 m s^–1 is typical for 6–9 μm diameter particles. Received: 17 February 1998 / Accepted in revised form: 17 June 1998     

Factors Influencing the Formation and Stability of Filled Hydrogel Particles Fabricated by Protein/Polysaccharide Phase Separation and Enzymatic Cross-Linking

Filled hydrogel particles can be used to encapsulate, protect, and deliver lipophilic components. In this study, we investigated the influence of preparation conditions on the size of filled hydrogel particles created using biopolymer phase separation and enzymatic cross-linking. We then investigated the stability of these particles to external stresses: pH (pH 2–8); heat (40°–90 °C, 20 min); sodium chloride (0–500 mM); and calcium chloride (0–8 mM). Filled hydrogel particles were fabricated as follows: (i) high methoxy pectin, sodium caseinate, and caseinate-coated lipid droplets were mixed at pH 7 under conditions where phase separation due to thermodynamic incompatibility occurred; (ii) this mixture was acidified (pH 5) to induce adsorption of anionic pectin molecules around lipid-filled caseinate-rich particles; (iii) the caseinate within the particles was enzymatically cross-linked using transglutaminase. Three mixing conditions (0, 100, and 1,000 rpm) were tested during particle acidification. Particle size measurements indicated that larger particles were formed at 0 and 100 rpm than at 1,000 rpm. Under high pH conditions (pH 6–8), particles cross-linked with transglutaminase remained intact while control particles (not cross-linked) disintegrated. The addition of calcium to both control and cross-linked particles resulted in system gelation above 4 mM calcium chloride. Control and cross-linked particles remained stable to heating and to the addition of sodium chloride. Results from this study demonstrate the versatility and robustness of this delivery system for lipophilic bioactives.     

Flow chamber analysis of size effects in the adhesion of spherical particles

The non-specific adhesion of spherical micro- and nano-particles to a cell substrate is investigated in a parallel plate flow chamber. Differently from prior in-vitro analyses, the total volume of the particles injected into the flow chamber is kept fixed whilst the particle diameter is changed in the range 0.5-10 microm. It is shown that: (i) the absolute number of particles adherent to the cell layer per unit surface decreases with the size of the particle as d(-1.7); (ii) the volume of the particles adherent per unit surface increases with the size of the particles as d(+1.3). From these results and considering solely non-specific particles, the following hypothesis are generated (i) use the smallest possible particles in biomedical imaging and (ii) use the largest possible particles in drug delivery.     

Mixing and propulsion of the contents of the reticulo-rumen]

Roughage intake and digestion by ruminants involve complex interactions between the roughage constituents, the microorganisms in the reticulo-rumen (RR) and its motility. Ruminal digestion requires intense activity, ie comminution of feed particles and mixing and propulsion of digesta. The regular repetition of the contraction sequences in the RR every 50 to 70 s subjects the digesta to a consistent pattern of movements. The particles are distributed according to their functional density which depends on the density of the plant structure of the particle, the liquid inside the particle and also the gas, ie on the degree of particle fermentation. An interwoven mat of large low-density particles fills the dorsal sac and the top of the ventral sac of rumen. This mat traps part of the small high density particles. Squeezed by the contractions, the interwoven mat acts like a filter and lets a liquid containing small particles of high density pass into the bottom of the ventral sac. This liquid then flows into the reticulum and passes through the reticulo-omasal orifice (ROO). Chewing during rumination reduces particle size, eliminates particle gas and aids in separating the low-density particles, which are less fermented, from the heavy residues. The outflow of digesta, made possible by the opening of the ROO during the second phase of the reticular contraction, is highly selective. The effluent does not contain particles greater than 2 mm in size in sheep and 4 mm in cattle. This is due to the buoyancy of the large particles in the reticulum, to the self-filtration of the digesta during the passage through the ROO and possibly to backflow from the omasum to the reticulum. Finally, RR motor activity, ie continuously mixing the digesta and monitoring the evacuation of gas and outflow of digesta, allows the homeostasis in the rumen necessary to microbial fermentation. The characteristics of the ingested particles, their rates of size reduction and density increase, the consistency of digesta and the intensity of the rumen wall stimulations are all factors which depend on the nature of feed and intake level. Via mechanisms which are not yet all well known and/or quantified, these factors act upon the efficiency of the mixing and propulsion of the reticulo-ruminal content and thus upon the retention time of the feed in the RR as well as its digestive utilisation.(ABSTRACT TRUNCATED AT 400 WORDS)     

Topography and functional information of plasma membrane

By using atomic force microscope (AFM), the topography and function of the plasmalemma surface of the isolated protoplasts from winter wheat mesophyll cells were observed, and compared with dead protoplasts induced by dehydrating stress. The observational results revealed that the plasma membrane of living protoplasts was in a state of polarization. Lipid layers of different cells and membrane areas exhibited distinct active states. The surfaces of plasma membranes were unequal, and were characterized of regionalisation. In addition, lattice structures were visualized in some regions of the membrane surface. These typical structures were assumed to be lipid molecular complexes, which were measured to be 15.8±0.09 nm in diameter and 1.9±0.3 nm in height. Both two-dimensional and three-dimensional imaging showed that the plasmalemma surfaces of winter wheat protoplasts were covered with numerous protruding particles. In order to determine the chemical nature of the protruding particles, living protoplasts were treated by proteolytic enzyme. Under the effect of enzyme, large particles became relatively looser, resulting that their width was increased and their height decreased. The results demonstrated that these particles were likely to be of protein nature. These protein particles at plasmalemma surface were different in size and unequal in distribution. The diameter of large protein particles ranged from 200 to 440 nm, with a central micropore, and the apparent height of them was found to vary from 12 to 40 nm. The diameter of mid-sized protein particles was between 40―60 nm, and a range of 1.8―5 nm was given for the apparent height of them. As for small protein particles, obtained values were 12―40 nm for their diameter and 0.7―2.2 nm for height. Some invaginated pits were also observed at the plasma membrane. They were formed by the endocytosis of protoplast. Distribution density of them at plasmalemma was about 16 pits per 15 μm2. According to their size, we classified the invaginated pits into two types―larger pits measuring 139 nm in diameter and 7.2 nm in depth, and smaller pits measuring 96 nm in diameter and 2.3 nm in depth. On dehydration-induced dead pro-toplasts, the degree of polarization of plasma membranes decreased. Lipid molecular layers appeared relatively smooth, and the quantity of integral proteins reduced a lot. Invaginated pits were still de-tectable at the membrane surface, but due to dehydration-induced protoplast contraction, the orifice diameter of pits reduced, and their depth increased. Larger pits averagely measuring 47.4 nm in di-ameter and 31.9 nm in depth, and smaller pits measuring 26.5 nm in diameter and 43 nm in depth at average. The measured thickness of plasma membranes of mesophyll cells from winter wheat examined by AFM was 6.6―9.8 nm, thicker in regions covered with proteins.     

No distinct difference in the excretion of large particles of varying size in a wild ruminant,the banteng (<Emphasis Type="Italic">Bos javanicus</Emphasis>)

The forestomach of ruminants and camelids does not only allow a differential excretion of fluids and small particles but also a differential excretion of small and large particles. The question whether larger particles of different size classes are also retained for different time periods, or whether simply a particle-size threshold exists above which all particles of a size higher than this threshold are retained in an undifferentiated manner, has not been addressed so far. We determined the mean retention time (MRT) of different-sized large particles (10 and 20 mm) in three banteng (Bos javanicus) on two forage only diets, grass and grass hay. We used cerium-mordanted fibre (10 mm) and lanthanum-mordanted fibre (20 mm) as particle markers, mixed in the food. Average total tract MRT for large and very large particles at the grass diet was 58 and 56 h and at the grass hay diet 66 and 64 h, respectively. Very large particles moved slightly faster than large particles through the gut of the banteng. Three interpretations are possible: Very large particles are resubmitted to rumination sooner than large particles; ingestive mastication of the particle markers could have reduced the difference in the size of the particle markers; alternatively, particle retention may be governed by a threshold, above which all particles of a size higher than this threshold are retained in an undifferentiated manner. In order to test these possibilities, experiments with fistulated animals would have to be performed.     

Particle classification from light scattering with the scanning flow cytometer.

BACKGROUND:The differential light-scattering pattern, an indicatrix, provides the most complete characterization of the optical properties of a particle. Particle classification can be performed on the basis of particle parameters retrieved from the indicatrices. This classification extends the ability of flow cytometry in particle recognition. METHODS:The scanning flow cytometer (SFC) permits an acquisition of traces of light scattering signals, i.e., native SFC traces, from single particles. The acquired native SFC traces are transformed into indicatrices. The performance of the SFC in measurements of indicatrices has been demonstrated for the following particles: lymphocytes, erythrocytes, polystyrene particles, and milk-fat particles. RESULTS:The structure and profile of the indicatrix for each particle type have been found to be unique. Classification of polystyrene particles has been performed on the basis of the map formed by particle refractive index and size. The polystyrene particles were classified using this map into different size categories ranging from 1.4-7 microm, with a size deviation of 0.07 microm. CONCLUSIONS:The method based on analysis of native SFC traces shows better performance in particle classification than the method based on the particle refractive index and size map. The classification performance of the SFC will be useful, for example, for particle sorting and particle identification, and with additional fluorescent measurements may have applications in multiparameter particle-based immunoassay.     

Novel changes in discoidal high density lipoprotein morphology: a molecular dynamics study

ApoA-I is a uniquely flexible lipid-scavenging protein capable of incorporating phospholipids into stable particles. Here we report molecular dynamics simulations on a series of progressively smaller discoidal high density lipoprotein particles produced by incremental removal of palmitoyloleoylphosphatidylcholine via four different pathways. The starting model contained 160 palmitoyloleoylphosphatidylcholines and a belt of two antiparallel amphipathic helical lipid-associating domains of apolipoprotein (apo) A-I. The results are particularly compelling. After a few nanoseconds of molecular dynamics simulation, independent of the starting particle and method of size reduction, all simulated double belts of the four lipidated apoA-I particles have helical domains that impressively approximate the x-ray crystal structure of lipid-free apoA-I, particularly between residues 88 and 186. These results provide atomic resolution models for two of the particles produced by in vitro reconstitution of nascent high density lipoprotein particles. These particles, measuring 95 angstroms and 78 angstroms by nondenaturing gradient gel electrophoresis, correspond in composition and in size/shape (by negative stain electron microscopy) to the simulated particles with molar ratios of 100:2 and 50:2, respectively. The lipids of the 100:2 particle family form minimal surfaces at their monolayer-monolayer interface, whereas the 50:2 particle family displays a lipid pocket capable of binding a dynamic range of phospholipid molecules.     

The interaction of surface and dust particle size on the pick-up and grooming behaviour of the German cockroachBlattella germanica

The relationship between the particle size of an inert silica dust, its up-take from different surfaces and the grooming behaviour of males, gravid females, and fifth and sixth instar nymphs of the German cockroachBlattella germanica (Dictyoptera: Blattellidae) (L.) was investigated. The normal grooming behaviour of gravid females, nymphs and males differed according to sex and age. The gravid females and nymphs exhibited greater grooming activity than the males, especially of the antennae and the legs. Gravid females, nymphs, and adult males exhibited increased grooming activity after exposure to dust in the size range 0.5–63 μm, but there was no significant difference in grooming behaviour from the control when cockroaches were exposed to dust sizes greater than 70 μm. Antennal grooming by males was greater than leg grooming when exposed to all dust sizes, except size particles ranging 4.5–7.5 μm. A dust pick-up experiment indicated that the average amount of dust transferred toB. germanica is affected by particle size, the porosity of the treated surface, and the sex and age of the cockroaches. Gravid females picked up greater amounts of dust than fifth and sixth instar nymphs, which in turn picked up more dust than males. Silica dust particles (0.5–7.5 μm) were picked up more effectively than larger particle sizes, by all three categories, males, females and fifth and sixth instar nymphs of cockroach on all three test surfaces plastic, ceramic and unpainted plywood. Plywood was the least effective surface for transfer of dust, of all sizes, to males, females and nymphs.