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PubMed1992

Adaptations in skeletal muscle capillarity following changes in oxygen supply and changes in oxygen demands.

The effects of changes in oxygen supply and oxygen demands on fiber cross-sectional areas, capillary densities and capillary to fiber ratios were determined in three skeletal muscles of rat. The muscles examined were the vastus lateralis, soleus, and diaphragm. Reduced oxygen supply was produced by subjecting rats to ambient hypoxia, and increased oxygen demands were produced by subjecting rats to low ambient temperatures or treatment with thyroxin. Capillaries were visualized by injecting fluorescent dyes into the circulation. Muscles were quick frozen at resting lengths to preserve normal fiber geometry and were subsequently sectioned on a cryostat. All of the muscles sampled from animals in the experimental groups had elevated capillary densities. However, capillary to fiber ratios were not increased significantly in any muscle, for any experimental condition. Thus, all of the observed differences in capillarity were due to changes in the intrinsic rate of muscle fiber growth. Further, the relations of capillary density and capillary to fiber ratio to fiber area were the same as those obtained during normal maturation, suggesting that capillary growth is closely linked to the intrinsic rate of fiber growth.

Adaptation, Physiological↗
PubMed1994

Prediction of mechanical damage to animal cells in turbulence.

In previous work a model was proposed for estimation of disruption of animal cells in turbulent capillary flows using information about the hydrodynamics, and cell mechanical properties determined by micromanipulation. The model assumed that the capillary flow consists of a laminar sublayer and a homogeneous turbulent region, and within the latter eddies of sizes similar to or smaller than the cells interact with those cells, causing local surface deformations. The proposed mechanism of cell damage was that such deformations result in an increase in membrane tension and surface energy, and that a cell disrupts when its bursting membrane tension and bursting surface energy are exceeded. The surface energy of the cells was estimated from the kinetic energy of appropriate sized eddies. To test the model, cells were disrupted in turbulent flows in capillaries at mean energy dissipation rates ranging from 800 to 2 x 10(4) Wkg-1. The model assumed that the specific lysis rate is almost independent of the number of passes, which was verified by the experimental data. The implication was that despite the damage the cell mechanical properties did not change markedly during multiple recirculations through the capillaries. On average the model underestimated the cell disruption by about 15%. Although the model gave reasonably good predictions, it lacks proper explanation of the independence of the specific lysis rate on the number of passes. In this paper it is shown that this problem can be resolved in principle by consideration of the localisation of the energy dissipation in turbulent capillary flows. The necessity of further modelling of cell-turbulence interactions is demonstrated.

Animals↗
PubMed1984

Bacterial growth in artificial capillary spaces.

In experiments on the influence of microscopic capillaries on the growth of Escherichia coli, Brevibacterium linens, Micrococcus flavus and Bacillus cereus, a distinct delay in growth was observed. The difference in counts was greatest after 8 h. Later a slight equilibration of counts was noted. With B. cereus, only slight or no spore formation was observed under microcapillary conditions.

Bacillus cereus↗
PubMed1995

Separation of gamma- and alpha-linolenic acid containing triacylglycerols by capillary supercritical fluid chromatography.

The separation of gamma- and alpha-linolenic acid containing triacylglycerols with an identical acyl carbon number and degree of unsaturation was obtained on capillary supercritical fluid chromatography using a 25% cyanopropyl-75% methylpolysiloxane stationary phase. The resolution of 1,3-dioleoyl-2-alpha-linolenoyl-sn-glycerol was 1.35 on a 10 m x 50 microns i.d. column, whereas the resolution was enhanced to 1.66 by combining two 10-meter columns in series. The difference in the position of double bonds in one linolenic acid residue of triacylglycerols resulted in two series of peaks in the separation of alpine currant (Ribes alpinum) and black currant (R. nigrum) seed oils. The use of the 10-meter column was found to be appropriate for the screening of the triacylglycerol profile in both seed oils studied.

Capillary Action↗
PubMed1995

Determination of lysine modification product epsilon-N-pyrrolylnorleucine in hydrolyzed proteins and trout muscle microsomes by micellar electrokinetic capillary chromatography.

epsilon-N-Pyrrolylnorleucine (Pnl) is a product of the reaction between the lipid peroxidation product 4,5(E)-epoxy-2(E)-heptenal (EH) and the epsilon-amino group of lysine. Because Pnl might also be produced in proteins, a specific method to determine this compound in protein hydrolysates has been developed. Homoarginine, added as the internal standard, and Pnl are derivatized with diethyl ethoxymethylenemalonate and analyzed by micellar electrokinetic capillary chromatography. The method also analyzes lysine and arginine, and these analyses were useful in determining losses of these amino acids after treatment with EH. The lowest concentration of Pnl detected with acceptable reproducibility is 5 nmol/mL, and the coefficient of variation was determined from four standard curves assayed on separate days. Detector response was linear for samples containing 1.6 to 74 nmol/mL of Pnl. The assay was applied in investigations of Pnl production in bovine serum albumin (BSA) and trout muscle microsomes treated with EH. When BSA was incubated overnight with 30 mM EH, 76% of lysine residues were modified, and a part of these residues were detected as Pnl (12%). Pnl formation was also detected when trout muscle microsomes were incubated for three hours with 1 or 10 mM EH. These results show that Pnl is produced in vitro in proteins treated with the lipid peroxidation product EH, and suggest that Pnl might also be constituent of in vivo damaged proteins by their reaction with oxidized lipids.

Aldehydes↗
PubMedDate not supplied

Capillary versus aspiration biopsy: effect of needle size and length on the cytopathological specimen quality.

PURPOSE: To test the value of the nonaspiration, or capillary, biopsy technique by experimental comparison with the conventional fine-needle aspiration technique using various needle gauges and lengths. METHODS: On fresh hepatic and renal tissue from five autopsies, multiple biopsy specimens were taken with 20, 22, and 23-gauge Chiba needles of 5, 10, 15, and 20-cm length, using the aspiration technique and the capillary technique. The resultant specimens were graded on the basis of a grading scheme by a cytopathologist who was blinded to the biopsy technique. RESULTS: The capillary technique obtained less background blood or clot which could obscure diagnostic tissue, although not significantly different from the aspiration technique (p = 0.2). However, for the amount of cellular material obtained, retention of appropriate architecture, and mean score, the capillary technique performed statistically worse than aspiration biopsy (p < 0.01). In addition, with decreasing needle caliber (increasing needle gauge) and increasing length, the capillary biopsy was inferior to the aspiration biopsy. CONCLUSION: The capillary biopsy technique is inferior to the aspiration technique according to our study. When the capillary technique is to be applied, preference should be given to larger caliber, shorter needles.

Biopsy, Needle↗
PubMed1998

Quantifying the cleanliness of glass capillaries.

I used capillary rise methods to investigate the lumenal surface properties of quartz (fused silica, Amersil T-08), borosilicate (Corning 7800), and high-lead glass (Corning 0010) capillaries commonly used to make patch pipets. I calculated the capillary rise and contact angle for water and methanol from weight measurements. The capillary rise was compared with the theoretical maximum value calculated by assuming each fluid perfectly wetted the lumenal surface of the glass (i.e., zero contact angle, which reflects the absence of surface contamination). For borosilicate, high-lead, and quartz capillaries, the rise for water was substantially less than the theoretical maximum rise. Exposure of the borosilicate, lead, and quartz capillaries to several cleaning methods resulted in substantially better--but not perfect--agreement between the theoretical maximum rise and calculated capillary rise. By contrast, the capillary rise for methanol was almost identical in untreated and cleaned capillaries, but less than its theoretical maximum rise. The residual discrepancy between the observed and theoretical rise for water could not be improved on by trying a variety of cleaning procedures, but some cleaning methods were superior to others. The water solubility of the surface contaminants, deduced from the effectiveness of repeated rinsing, was different for each of the three types of capillaries examined: Corning 7800 > quartz > Corning 0010. A surface film was also detected in quatz tubing with an internal filament. I conclude that these borosilicate, quartz, and high-lead glass capillaries have a film on the lumenal surface, which can be removed using appropriate cleaning methods. The surface contaminants may be unique to each type of capillary and may also be hydrophobic. Two simple methods are presented to quantitate the cleanliness of glass capillary tubing commonly used to make pipets for studies of biological membranes. It is not known if the surface film is of importance in electrophysiological studies of biological membranes.

Capillary Action↗
PubMedDate not supplied

Diffusion and transfer of antibody proteins from a sugar-based hydrogel.

Diffusion of antibody protein from hydrogel films and hydrogel encapsulated in a microcapillary was studied. Thin hydrogel films were formed by crosslinking 6-acryloyl-B-O-methylgalactoside with N,N'-methylene-bis-acrylamide and the diffusive transport of monoclonal antimouse IgG-FITC into and out of the hydrate films was measured. Diffusion coefficients in 2 and 4% crosslinked hydrogel films were measured. The measured diffusion constants determined for IgG in both the 2 and 4% hydrogel films were comparable to the free diffusion of IgG in bulk water (Dmean approximately 10(-7) cm2/s). In addition, 2% crosslinked hydrogels were prepared in a capillary tube and the transport of antimouse IgG-FITC into and out of the hydrated hydrogel was measured. Kinetic analysis indicated that the protein transport through the capillary hydrogel was faster than would be expected for a simple diffusion process. Finally, by utilizing the diffusion of antibody from the capillary hydrogel, transfer of antibody to a silica surface was demonstrated. A capillary hydrogel loaded with antimouse IgG-FITC was used to transfer the protein to a silica surface forming a 30-micron spot of antibody, which was imaged using fluorescence microscopy. These results may lead to the development of a nonlithographic method of patterning antibodies on surfaces for use in integrated microimmunosensors.

Acrylamides↗
PubMed2005

Functional and histological effects of intravaginal electrical stimulation on the pelvic muscles: a study in the rat.

We studied functional and histological effects of electrical stimulation (ES) on pelvic muscles of the rat. With intravaginal electrodes, the musculus pubococcygeus and musculus iliococcygeus in the awake animal were stimulated three times 6 min per day with 5 min of rest in between, 5 days per week, 7 consecutive weeks with a biphasic rectangular symmetrical current of 25 Hz, 400-mus pulse duration, on/off time of 5/10 and with an amplitude of 2-4 mA. A "sham group" received the same handling but no stimulation. Contraction measured with intra-rectal pressure during stimulation increased more in the stimulated than in the sham group, but did not reach statistical significance probably due to low power. The 2A fast fibres increased with 14% in the musculus iliococcygeus and with 6% in the musculus pubococcygeus. Type 1 slow fibres did not change. Increased capillary density was found after stimulation. Repeated intravaginal ES has mainly an influence on the fast fibres in the pelvic muscles. To influence slow fibres, another stimulation program or current parameters would seem necessary.

Adenosine Triphosphatases↗
PubMed2002

Stir bar sorptive extraction-thermal desorption-capillary GC-MS applied to biological fluids.

A new sample preparation method, stir bar sorptive extraction (SBSE), has been evaluated for the enrichment of organic solutes from biological fluids such as urine and blood. In SBSE, a stir bar coated with a polydimethylsiloxane layer is stirred for a given time in the sample. After sampling the stir bar is placed in a thermal desorption unit coupled on-line to capillary gas chromatography-mass spectrometry (SBSE-TD-CGC-MS). The principle and operation of SBSE are presented. Total profiling and target compound analysis have been selected as applications to illustrate the performance of SBSE-TD-CGC-MS (MSD). It is demonstrated that a variety analytes ranging from biological markers (phenols, hormones, fatty acids) to artificial contaminants (recreational drugs, plasticizers) can be enriched with high sensitivity. For polar solutes, in-situ derivatization can enhance both recovery into the polydimethylsiloxane (PDMS) layer and chromatographic analysis. Two types of derivatization have been applied, derivatization with ethyl chloroformate and with acetic acid anhydride. Linearity, detectability, and repeatability are illustrated by the determination of 1-hydroxypyrene in a urine sample from a smoker.

Body Fluids↗
PubMed2002

Miniaturized solid-phase extraction as a sample preparation technique for the determination of phthalates in water.

Miniaturized solid-phase extraction (SPE) has been developed and successfully employed for the determination of organic species in water samples by liquid chromatography (LC). The method is based on the concept of a microscale extraction technique using a fused-silica capillary column for gas chromatography (GC), so-called in-tube solid-phase microextraction (SPME). The extraction conditions, such as the extraction time and flow-rate for the extraction and desorption process, were investigated as well as the effect of the internal structure of the extraction capillary on the efficiency. By inserting a stainless steel wire into the extraction capillary to reduce the internal volume of the capillary with the same surface area of the coating, an improved extraction and pre-concentration effects were obtained. Further pre-concentration was accomplished by the extraction device with a novel fiber-in-tube configuration. The direct coupling of the extraction method with a LC system has made it possible to determine low levels of phthalates in water samples without high consumption of organic solvents. The system developed must have potential applications for the analysis of environmental and biological samples in aqueous sample matrices.

Capillary Action↗
PubMed2002

On-line coupling of miniaturized solid-phase extraction and microcolumn liquid-phase separations.

On-line coupling of miniaturized solid-phase extraction methods and microcolumn separation techniques are reviewed. A conventional solid-phase micro-extraction device and fiber-in-tube solid-phase extraction was employed as the sample preparation method. In the fiber-in-tube technique, a novel fibrous polymeric material was introduced as the extraction medium for the microscale sample preparation step which could be directly coupled with microcolumn liquid-phase separation systems, such as microcolumn liquid chromatography, capillary electrophoresis, and capillary electrochromatography. The applications of these hyphenated systems, consisting of the sample preparation and separation processes, are also reviewed.

Capillary Action↗
PubMed2003

Trace determination of peptides in water samples using packed capillary liquid chromatography with UV and MS detection and characterization of peptide oxidation products by MS.

A capillary liquid chromatographic column switching method has been developed for fast and sensitive determination of peptides in water samples. Sample volumes of 1 mL were loaded onto a (320 microm I.D. x30 mm) 10 microm Kromasil C(18) pre-column, providing on-line analyte enrichment, prior to back-flushed elution onto a (320 microm I.D. x150 mm) 3.5 microm Kromasil C(18) analytical column. Loading flow rates of 250 microL/min and a mobile phase composition of acetonitrile/water/trifluoroacetic acid (22/77.9/0.1, v/v) provided a total analysis time of less than 25 minutes for the test peptides angiotensin II, bombesin, bradykinin, corazonin, neurotensin and substance P, using temperature programmed elution. In addition, solvent gradient elution and combined solvent gradient elution and temperature programming were explored. Using on-capillary UV detection at 210 nm resulted in a concentration limit of detection (cLOD) of about 1 ng/mL. The method was validated over the concentration range 1-100 ng/mL, yielding a coefficient of correlation of 0.997 or better. The within-assay ( n=6) and between-assay ( n=6) precisions of peak areas were on average 6% RSD and 5% RSD, respectively. When the method was applied to spiked chlorinated tap water samples, it was found that peptides containing methionine, tryptophan and cystine were oxidized. Identification of the oxidation products of the peptides in hypochlorite-treated water was done with positive electrospray ionization time-of-flight mass spectrometric detection.

Capillary Action↗
PubMed2000

Micropatterned solid-supported membranes formed by micromolding in capillaries.

The formation of individually addressable micropatterned solid-supported lipid bilayers has been accomplished by means of micromolding in capillaries. Small unilamellar vesicles were spread on glass slides to form planar supported membranes along microscopic capillaries molded as trenches into a polydimethylsiloxane (PDMS) elastomer. PDMS provides an elastic and transparent carrier for microcapillaries molded from silicon wafers displaying the desired inverse trenches. The so-called master structure has been conventionally etched into silicon by photolithography. The cured PDMS elastomer was briefly exposed to an oxygen plasma, rendering the surface hydrophilic, and subsequently attached to a glass surface in order to form hydrophilic capillaries equipped with flow-promoting pads on either side. One flowpad acts as a reservoir to be filled with the vesicle suspension, while the other one serves as a collector to ensure a sufficient capillary flow to cover the substrate completely. Formation of planar lipid bilayers on the glass slide along the capillaries was followed by imaging the flow and spreading of fluorescently labeled DMPC liposomes with confocal laser scanning microscopy. By means of scanning force microscopy in aqueous solution the formed lipid structures were identified and the height of the lipid bilayers was accurately determined. With both techniques, it was shown that the patterned bilayers remain separated and persist for several hours on the substrate in aqueous solution.

Biocompatible Materials↗
PubMed2000

Surface tension and viscosity of surfactant from the resonance of an oscillating drop.

The oscillating drop surfactometer (ODS) measures surface tension (gamma) and energy dissipation (damping constant b) of surfactant on a 1 microl sample. gamma is obtained from the period of oscillation and b from its free decay or from the phase shift slope in resonance. After calibration with substances with different gamma, corrections were made for capillary fixation and loss of mass by evaporation. Surface active substances are delivered from liposomes in the interior (subphase) or injected from outside, with microdrops (180 pl each) of solution. As an application example, we have investigated surfactant extract and pure phospholipid. In minutes after formation of a drop containing a diluted Survanta suspension, gamma, decreases by 20 mN/m, while b increases three-fold. This effect, assigned to spontaneous adsorption from liposomes to the surface, is not seen with pure dipalmitoylphosphatidylcholine (DPPC) under our conditions. However, microdrop injection of DPPC triggers a rapid decrease of gamma and a delayed strong increase in b. The effect is modulated by DPPC in the subphase and by cholesterol. Investigations with L-alpha-lysophosphatidylcholine show the high sensitivity of the ODS technique in the determination of the energy dissipation at air-liquid boundary surfaces. Although the ODS is limited to applications with gamma > 15 mN m(-1), it offers the advantage to give, with small samples and within seconds, a simultaneous readout of both Surface tension gamma and the parameter b, as a measure of surface viscosity.

1,2-Dipalmitoylphosphatidylcholine↗