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At least 19 recordsLinked to original sources

PubMed1979

Concentration of poliovirus from tap water using positively charged microporous filters.

Microporous filters that are more electropositive than the negatively charged filters currently used for virus concentrations from water by filter adsorption-elution methods were evaluated for poliovirus recovery from tap water. Zeta Plus filters composed of diatomaceous earth-cellulose-"charge-modified" resin mixtures and having a net positive charge of up to pH 5 to 6 efficiently adsorbed poliovirus from tap water at ambient pH levels 7.0 to 7.5 without added multivalent cation salts. The adsorbed virus were eluted with glycine-NaOH, pH 9.5 to 11.5. Electropositive asbestos-cellulose filters efficiently adsorbed poliovirus from tap water without added multivalent cation salts between pH 3.5 and 9.0, and the absorbed viruses could be eluted with 3% beef extract, pH 9, but not with pH 9.5 to 11.5 glycine-NaOH. Under water quality conditions in which poliovirus recoveries from large volumes of water were less than 5% with conventional negatively charged filters and standard methods, recoveries with Zeta Plus filters averaged 64 and 22.5% for one- and two-stage concentration procedures, respectively. Electropositive filters appear to offer distinct advantages over conventional negatively charged filters for concentrating enteric viruses from water, and their behavior tends to confirm the importance of electrostatic forces in virus recovery from water by microporous filter adsorption-elution methods.

Adsorption↗

PubMed1980

Rapid concentration of bacteriophages from large volumes of freshwater: evaluation of positively charged, microporous filters.

Microporous, positively charged Zeta Plus 60 S filters were evaluated for bacteriophage recovery from large volumes of natural water. A variety of phages adsorbed efficiently to the filter medium at water pH levels below neutrality, but adsorption was reduced above pH 7. Adsorbed phages were easily eluted with an arginine/1% beef extract solution, pH 9.0. A concentration procedure suitable for the field isolation of bacteriophages from large volumes of river water was devised. The procedure involves, (1) prefiltration through 10" cartridge depth filters, (2) adjustment of water pH to pH 5.5--6.0, (3) adsorption of the phages on to Zeta Plus 60 S filters, (4) elution of bound phage in a small volume of eluent, (5) secondary concentration by ultrafiltration of the resulting eluates. Using this procedure, bacteriophages in 65 1 of river water was concentrated to 35 ml with recoveries in the range 50--60%.

Adsorption↗

PubMed1983

Simplified micropore filter assay of neutrophil migration using whole blood.

A new method of micropore filter assay of neutrophil migration requiring only 0.1 ml of whole blood is described and compared with the standard separated polymorphonuclear neutrophil micropore filter assay. Whole blood was added to the upper compartment of a modified Boyden chemotactic chamber, and the neutrophils were allowed to migrate into a cellulose nitrate micropore filter. Acetic acid was used to remove erythrocytes and hemoglobin from the filter. Neutrophil chemotaxis was performed with cells from 19 neonates and 34 adults. The mean neonatal PMN migration was 40% of the adult value (48.1 +/- 12.6 vs 96.8 +/- 16.8 micron) with the whole blood assay and 39% of the adult value (44.7 +/- 10.1 vs 72.9 +/- 22.2 micron) with the separated polymorphonuclear neutrophil assay. The whole blood micropore filter assay, a simple and reliable method of determining neutrophil migration, is especially useful in studying patients when there are difficulties in obtaining large blood samples, such as with neonates and young children.

Cells, Cultured↗

PubMed1988

Novel approach for modifying microporous filters for virus concentration from water.

Electronegative microporous filters composed of epoxyfiberglass (Filterite) were treated with cationic polymers to enhance their virus-adsorbing properties. This novel and inexpensive approach to microporous filter modification entails soaking filters in an aqueous solution of a cationic polymer such as polyethyleneimine (PEI) for 2 h at room temperature and then allowing the filters to air dry overnight on absorbent paper towels. PEI-treated filters were evaluated for coliphage (MS2, T2, and phi X174) and enterovirus (poliovirus type 1 and coxsackievirus type B5) adsorption from buffer at pH 3.5 to 9.0 and for indigenous coliphages from unchlorinated secondary effluent at ambient pH. Adsorbed viruses were recovered with 3% beef extract (pH 9). Several other cationic polymers were used to modify epoxyfiberglass filters and were evaluated for their ability to concentrate viruses from water. Zeta potentials of disrupted filter material indicated that electronegative epoxyfiberglass filters were made more electropositive when treated with cationic polymers. In general, epoxyfiberglass filters treated with cationic polymers were found to adsorb a greater percentage of coliphages and enteroviruses than were untreated filters.

Coliphages↗

PubMed1999

Human endothelial cells cultured on microporous filters used for leukocyte transmigration studies form monolayers on both sides of the filter.

A growing number of studies on the mechanism of leukocyte transendothelial migration use endothelial cells grown on microporous filters as an in vitro model of endothelium. Ultrastructural examination of such a model system previously demonstrated that human pulmonary artery endothelial cells (HPAEC) formed confluent monolayers on both sides of the 3-microm-pore filter (Mackarel et al., 1999). To determine whether this was a characteristic specific to pulmonary artery endothelial cells, the growth characteristics of a human pulmonary microvascular endothelial cell type (HMVEC-L) and the widely used human umbilical vein endothelial cells (HUVEC) on 3-microm microporous filters were examined by transmission electron microscopy (TEM). Similar to HPAEC, HMVEC-L and HUVEC were also found to grow on both sides of the filter. All three endothelial cell types were capable of migrating through the 3 microm pores of the filter to form a monolayer on the filter underside. The endothelial cells on the underside were orientated in an inverted position with the luminal surface facing away from the filter. Such 'bilayer' formation was observed at a range of seeding densities and in different culture media. Despite the presence of a bilayer of endothelial cells, TEM demonstrated that neutrophils migrated successfully across the cell-filter-cell system. Previous transmigration reports in which an in vitro model similar to ours was used have often assumed only one layer of endothelial cells. The observations reported here indicate that while endothelial cells on microporous filters are useful models for examining leukocyte-endothelial interactions, they are not appropriate for studies examining endothelial cell 'sidedness.'

Cell Culture Techniques↗

PubMed1987

Characterization of retinal pigment epithelial cells cultured on microporous filters.

Retinal pigment epithelium (RPE) cultured on microporous filter supports was compared to RPE cultured on plastic and evaluated for features characteristic of RPE in vivo. RPE cells grown on filters were cuboidal, formed junctional complex structures between cells, and had elaborate microvilli and basal infoldings similar to RPE in vivo, while RPE grown on plastic also formed intercellular junctions but appeared squamous and had few microvilli and basal infoldings. RPE grown on filters or plastic secreted an extracellular matrix at the basal surface and ingested isolated rat rod outer segments at the apical surface. RPE grown on filters coated with laminin or fibronectin became confluent more rapidly than RPE grown on uncoated filters, while RPE grown at the same density on filters coated with collagen type I did not become confluent. The laminin and fibronectin coatings did not alter the RPE cell morphology; however, cells seeded on collagen-coated filters grew in large disorganized clusters. RPE grown on laminin-coated filters formed functional tight junctions as evidenced by the capacity of RPE monolayers to prevent the bulk flow of medium and the passage of trypan blue across the filter. Radiolabeled sucrose and inulin were used to measure the paracellular flux through the tight junctions between cells. The passage of these tracers was linear over time, with the lower molecular weight tracer, sucrose, passing through the monolayer more readily than inulin. Values for the flux of radiolabeled bovine serum albumin across RPE monolayers fell between values for sucrose and inulin. The results from these studies show that RPE monolayers cultured on laminin-coated filters maintain a morphology similar to that of RPE in vivo, are capable of ingesting rod outer segments, and form a selectively permeable barrier to various tracers. This culture system should be useful for studies of transepithelial transport, secretion, endocytosis and exocytosis that require independent control of the extracellular environment at the apical and basolateral cell surfaces.

Animals↗

PubMed1985

Effects of bentonite clay solids on poliovirus concentration from water by microporous filter methods.

To determine whether suspended solids interfere with enteric virus recovery from water by microporous filter methods, the effects of bentonite clay solids at a concentration of 10 nephelometric turbidity units on the recovery of poliovirus type 1 from seeded, activated carbon-treated, filtered tap water were studied. Volumes (500 ml) of virus-laden water at pH 5.5 or 7.5, with and without 50 mM MgCl2, were filtered through 47-mm-diameter, electropositive (Virosorb 1MDS) and electronegative (Filterite) filters that had been pretreated with Tween 80 to minimize direct virus adsorption to filter surfaces. Bentonite solids enhanced virus retention on both types of filters, even under conditions in which viruses were not solids associated. However, bentonite solids also interfered with elution of retained viruses when eluting with 0.3% beef extract-50 mM glycine (pH 9.5). Under some conditions, overall virus recoveries were lower from water with bentonite solids than from solids-free control water. The results of this study indicate that clay turbidity can interfere somewhat with virus recovery by current microporous filter methods.

Adsorption↗

PubMed2000

Influence of salts on virus adsorption to microporous filters.

We investigated the direct and indirect effects of mono-, di-, and trivalent salts (NaCl, MgCl(2), and AlCl(3)) on the adsorption of several viruses (MS2, PRD-1, phiX174, and poliovirus 1) to microporous filters at different pH values. The filters studied included Millipore HA (nitrocellulose), Filterite (fiberglass), Whatman (cellulose), and 1MDS (charged-modified fiber) filters. Each of these filters except the Whatman cellulose filters has been used in virus removal and recovery procedures. The direct effects of added salts were considered to be the effects associated with the presence of the soluble salts. The indirect effects of the added salts were considered to be (i) changes in the pH values of solutions and (ii) the formation of insoluble precipitates that could adsorb viruses and be removed by filtration. When direct effects alone were considered, the salts used in this study promoted virus adsorption, interfered with virus adsorption, or had little or no effect on virus adsorption, depending on the filter, the virus, and the salt. Although we were able to confirm previous reports that the addition of aluminum chloride to water enhances virus adsorption to microporous filters, we found that the enhanced adsorption was associated with indirect effects rather than direct effects. The increase in viral adsorption observed when aluminum chloride was added to water was related to the decrease in the pH of the water. Similar results could be obtained by adding HCl. The increased adsorption of viruses in water at pH 7 following addition of aluminum chloride was probably due to flocculation of aluminum, since removal of flocs by filtration greatly reduced the enhancement observed. The only direct effect of aluminum chloride on virus adsorption that we observed was interference with adsorption to microporous filters. Under conditions under which hydrophobic interactions were minimal, aluminum chloride interfered with virus adsorption to Millipore, Filterite, and 1MDS filters. In most cases, less than 10% of the viruses adsorbed to filters in the presence of a multivalent salt and a compound that interfered with hydrophobic interactions (0.1% Tween 80 or 4 M urea).

Adsorption↗

PubMed2000

Dendritic cell migration in different micropore filter assays.

Dendritic cells (DC) are highly motile and have been shown to migrate in vitro or in vivo towards various chemoattractants. Micropore filter methods with polycarbonate filters are generally used in these in vitro experiments. Among others, the main drawback of these filters is their thickness, which does not allow any assessment of effects of absolute concentration compared to gradients. The aim of this study was to establish a chemotaxis assay for dendritic cells using nitrocellulose filters, which can be adapted for checkerboard studies to distinguish between chemokinesis and chemotaxis. Immature DC were generated by culture of peripheral blood mononuclear cells using granulocyte-macrophage colony-stimulating factor and interleukin-4. We tested cell migration into nitrocellulose in a Boyden microchemotaxis chamber (leading front assay) and compared this method to the commonly used polycarbonate filter technique. Dendritic cells migrated well into nitrocellulose towards gradients of formyl peptide, complement fragment 5a, and monocyte chemotactic protein-3. The nitrocellulose method appeared to be more sensitive as compared to experiments testing migration across polycarbonate filters. Subsequent checkerboard analyses confirmed chemotactic activities of formyl peptide and complement fragment 5a. However, depending on the assay system, chemotaxis in polycarbonate filters but chemokinesis in nitrocellulose filters were observed for monocyte chemotactic protein-3. Measurement of DC migration in a cellulose nitrate micropore filter assay is more sensitive than the commonly used polycarbonate method and can be adapted for checkerboard analyses.

Chemokine CCL7↗

PubMed2002

Improved method for recovery of bacteriophage from large volumes of water using negatively charged microporous filters.

Current virus-recovery procedures using negatively charged microporous filters provide an inexpensive, reliable method for the recovery and detection of enteroviruses from water and wastewater; however, adjustment of the test samples to pH 3.5 to promote enterovirus adsorption results in significant inactivation of bacteriophage and an inability to simultaneously recover them from large volumes of water using this procedure. Procedures specifically designed for the detection of bacteriophage are currently in use but generally are only effective for small volumes of water. Positively charged filters can be used to recover both enteroviruses and bacteriophage from large volumes of water at neutral pH; however, the filters are expensive. The addition of manganese chloride to test solutions at pH 3.5 prior to filtration through negatively charged Filterite filters allowed for sampling of larger volumes of water by reducing the inactivation of bacteriophage and increasing the recovery of PRD1, MS2, and naturally isolated bacteriophage by a factor of four or five when compared with recoveries from solutions without MnCl2. This method provides an inexpensive, reliable alternative to large-volume bacteriophage recovery procedures that use positively charged filters at neutral pH.

Adsorption↗

PubMed2001

[Micropore filters for measuring red blood cell deformability and their pore diameters].

Micropore filters are the most important components in micropore filtration testes for assessing red blood cell (RBC) deformability. With regard to their appearance and filtration behaviors, comparisons are made for different kinds of filters currently in use. Nickel filters with regular geometric characteristics are found to be more sensitive to the effects of physical, chemical, especially pathological factors on the RBC deformability. We have critically reviewed the following viewpoint that filters with 3 microns pore diameter are more sensitive to cell volume than to internal viscosity while filters with 5 microns pore diameter are just the opposite. After analyzing the experiment results with 3 microns and 5 microns filters, we point out that filters with smaller pore diameters are more suitable for assessing the RBC deformability.

Erythrocyte Deformability↗

PubMed1995

An image processing procedure for the assessment of normality curves of motility of human granulocytes in micropore filters.

With the conventional methods used in current practice, the results of human granulocyte motility assays are expressed by a numerical value that defines the leading front of cells into micropore filters. The authors have developed a fast procedure which allows the complete curve of polymorphonuclear leukocyte (PMN) migration to be obtained, from the initial plane of a filter to the maximum propagation depth. In this way, more information is given than that expressed by a single parameter. The paper describes the procedure used to determine the normality curves both for random and stimulated migration, according to a simple model which defines the PMN propagation in micropore filters. A normality band that defines variations due to the stochastic process is also reported.

Cell Movement↗

PubMed1981

Neutrophil-endothelial cell interactions on endothelial monolayers grown on micropore filters.

We have developed a technique for growing endothelial monolayers on micropore filters. These monolayers demonstrate confluence by phase and electron microscopy and provide a functional barrier to passage of radiolabeled albumin. Neutrophils readily penetrate the monolayer in response to chemotaxin, whereas there is little movement in the absence of chemotaxin. This system offers unique advantages over available chemotaxis assays and may have wider applications in the study of endothelial function.

Blood Vessels↗

PubMed1985

Micropore filters for sterile filtration may leach toxic compounds affecting cell cultures (HL 60).

Reduction in HL-60 cell growth could be traced unexpectedly to sterile filtration procedures. To circumvent this problem, 11 available brands of micropore filters (five prepacked and six to be packed and autoclaved) were investigated with the aim of finding the least toxic product. Samples of 10 ml of RPMI 1640 medium with 10% fetal calf serum added were filtered through four filters of each brand to detect even small amounts of leached toxic compounds. HL-60 cells were cultured in these filtered media for three days and the results compared with cultures using unfiltered medium. A large variation in growth inhibition was found between the filters investigated, ranging from 0% to about 90%. The growth inhibition was due to leaching of toxic compounds, as revealed by viability test, reseeding, and direct microscopy. Adsorption of essential proteins to the micropore filters was found for only one of the filters investigated in this study.

Cell Survival↗

PubMed2001

[Collecting of trichloroaniline in the air by micropore filter membrane and determination by gas chromatography].

Trichloroaniline in the air was collected with micropore filter membrane and desorbed with cyclohexane, separated with a column OV-17, OV-210 and determined by GC-ECD. The materals for the filter of collection, solution of desorping and flow-rate of sampling were selected. The detectable limit is 0.01 mg/L. When the concentration of standard solutions is 1-10 mg/L, the relative standard deviation (RSD) is 4.5%-2.3%. There is a linear relation within the range of 0.005-30 mg/L. The sampling efficiency is 98.7%-100%. The samplers were stable for at least 15 days. This method is proved to be accurate, sensitive, fast and simple for sampling and carrying, and it is method is suitable for the determination of trichloroaniline in air, for personal detection and detection in the fields.

Air Pollutants↗

PubMed1995

Determination of RBC flow clogging via micropore filters.

Using a prototype system we determine the dynamic behaviour of the clogging effect of an erythrocyte suspension flow via micropore filters. The system functions as a simple electronic device and a characteristic curve of the change in height of the level of RBC suspension which flows through the filter versus time is plotted.

Erythrocytes↗

PubMed2001

[Determination of beryllium in atmospheric particulate by graphite furnace atomic absorption spectrophotometer using aluminium as a matrix modifier agent sampling with micropore filter membrane].

Beryllium in the atmospheric particulates is collected with 0.22 micron pore diameter micropore filter membrane, which is determined by graphite furnace atomic absorption spectrophotometer using aluminium as a matrix modifier agent. This sampling method shown several advantages: easy, complete and fast digestion; low membrane blank; high elution recovery, standard addition recovery, precision and accuracy. The addition of 800 mg.L-1 aluminium to beryllium solution allows ashing temperature of 2,000 degrees C. The detection limit of 13.5 ng was obtained.

Air Pollutants↗

PubMed1989

Further studies on the flow of aqueous humor through microporous filters.

It has recently been shown that aqueous humor is able to obstruct flow through Nuclepore polycarbonate filters having flow dimensions similar to those found in the juxtacanalicular tissue (JCT). We undertook studies designed to identify the component(s) of aqueous humor responsible for this obstruction and to determine the mechanism of blockage. We conclude that aqueous humor contains two components (one of which is specific to aqueous) which, when simultaneously present, hydrophobically bind to microporous filters and lead to filter blockage. Some implications for aqueous humor flow through the JCT are briefly discussed.

Animals↗