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Biomedical subjects

E R Scott

Publications and source records attributed to E R Scott.

16 recordsLinked to original sources

Electrically facilitated molecular transport. Analysis of the relative contributions of diffusion, migration, and electroosmosis to solute transport in an ion-exchange membrane.

Electrically facilitated molecular transport in an ion-exchange membrane (Nafion, 1100 equiv wt) has been studied using a scanning electrochemical microscope. The transport rates of ferrocenylmethyltrimethylammonium (a cation), acetaminophen (a neutral molecule), and ascorbate (an anion) through approximately 120-micron-thick membranes were measured as a function of the iontophoretic current passed across the membrane (-1.0 to +1.0 A/cm2). Transport rates were analyzed by employing the Nernst-Planck equation, modified to account for electric field-driven convective transport. Excellent agreement between experimental and theoretical values of the molecular flux was obtained using a single fitting parameter for each molecule (electroosmotic drag coefficient). The electroosmotic velocity of the neutral molecule, acetaminophen, was shown to be a factor of approximately 500 larger than that of the cation ferrocenylmethyltrimethylammonium, a consequence of the electrostatic interaction of the cation with the negatively charged pore walls of the ion-exchange membrane. Electroosmotic transport of ascorbate occurred at a negligible rate due to repulsion of the anion by the cation-selective membrane. These results suggest that electroosmotic velocities of solute molecules are determined by specific chemical interactions of the permeant and membrane and may be very different from the average solution velocity. The efficiency of electroosmotic transport was also shown to be a function of the membrane thickness, in addition to membrane/solute interactions.

Acetaminophen↗

Scanning electrochemical microscopy of iontophoretic transport in hairless mouse skin. Analysis of the relative contributions of diffusion, migration, and electroosmosis to transport in hair follicles.

Scanning electrochemical microscopy (SECM) is used to measure spatially localized diffusive and iontophoretic transport rates in hairless mouse skin. Molecular fluxes within individual hair follicles are quantified by measuring the rate at which redox-active probe molecules emerge from the follicle. The influence of an applied current on the flux of an anion (ascorbate), a cation (ferrocenylmethyltrimethylammonium), and a neutral molecule (acetaminophen) is used to determine the contributions of diffusion, migration, and electroosmosis to iontophoretic transport. The direction of electroosmotic transport is consistent with hair follicles possessing a net negative charge at neutral pH. Electroosmosis results in a modest increase in the transport rate of the neutral molecule (a factor of approximately 2.4x at an iontophoretic current density of 0.1 mA/cm(2)). Larger enhancements in the flux of the electrically charged species are associated with migration. The electroosmotic flow velocity within hair follicles is established to be 0.5 (+/-0.1) microm/s at 0.1 mA/cm(2), independent of the electrical charge of permeant. The net volume flow rate across skin resulting from electroosmosis in hair follicles is estimated to be 0.3 microL/cm(2)h. The results suggest that hair follicles are a significant pathway for electroosmotic solution flow during iontophoresis. The radius of the hair follicle openings in hairless mouse skin is measured to be 21 +/- 5 microm.

Algorithms↗

Visualization and analysis of electroosmotic flow in hairless mouse skin.

PURPOSE: To identify the physiological structures in hairless mouse skin responsible for the generation of electroosmotic flow during iontophoresis. Also, to determine the effects of changing the pH of the contacting solution on the magnitude of electroosmotic flow in these structures. METHODS: Localized diffusive and iontophoretic fluxes of a neutral molecule, hydroquinone (HQ), across hairless mouse skin were quantified using scanning electrochemical microscopy (SECM). The iontophoretic flux was determined as a function of the direction of the applied current and pH of the contacting solution. RESULTS: SECM images of HQ transport recorded during iontophoresis at moderate current densities (+/-0.1 mA/cm2) demonstrate that electroosmotic flow is localized to hair follicles. The direction of flow is from anode to cathode at pH > 3.5 and from cathode to anode at pH <3.5. CONCLUSIONS: Electroosmotic flow through hair follicles is an efficient and controllable means of transporting small, electrically neutral molecules across hairless mouse skin. Transport through the appendages is sensitive to the pH of the solution in contact with the skin. The isoelectric point of hair follicles, pI, is estimated to be 3.5 from the dependence of electroosmotic flow on the solution pH.

Animals↗

Origin of carbonate-magnetite-sulfide assemblages in Martian meteorite ALH84001.

A review of the mineralogical, isotopic, and chemical properties of the carbonates and associated submicrometer iron oxides and sulfides in Martian meteorite ALH84001 provides minimal evidence for microbial activity. Some magnetites resemble those formed by magnetotactic microorganisms but cubic crystals <50 nm in size and elongated grains <25 nm long are too small to be single-domain magnets and are probably abiogenic. Magnetites with shapes that are clearly unique to magnetotactic bacteria appear to be absent in ALH84001. Magnetosomes have not been reported in plutonic rocks and are unlikely to have been transported in fluids through fractures and uniformly deposited where abiogenic magnetite was forming epitaxially on carbonate. Submicrometer sulfides and magnetites probably formed during shock heating. Carbonates have correlated variations in Ca, Mg, and 18O/16O, magnetite-rich rims, and they appear to be embedded in pyroxene and plagiociase glass. Carbonates with these features have not been identified in carbonaceous chondrites and terrestrial rocks, suggesting that the ALH84001 carbonates have a unique origin. Carbonates and hydrated minerals in ALH84001, like secondary phases in other Martian meteorites, have O and H isotopic ratios favoring formation from fluids that exchanged with the Martian atmosphere. I propose that carbonates originally formed in ALH84001 from aqueous fluids and were subsequently shock heated and vaporized. The original carbonates were probably dolomite-magnesite-siderite assemblages that formed in pores at interstitial sites with minor sulfate, chloride, and phyllosilicates. These phases, like many other volatile-rich phases in Martian meteorites, may have formed as evaporate deposits from intermittent floods.

Carbonates↗

53Mn-53Cr dating of fayalite formation in the CV3 chondrite Mokoia: evidence for asteroidal alteration.

Fayalite grains in chondrules in the oxidized, aqueously altered CV3 chondrite Mokoia have large excesses of radiogenic chromium-53. These excesses indicate the in situ decay of short-lived manganese-53 (half-life = 3.7 million years) and define an initial 53Mn/55Mn ratio of 2.32 (+/-0.18) x 10(-6). This ratio is comparable to values for carbonates in CI and CM chondrites and for several classes of differentiated meteorites. Mokoia fayalites formed 7 to 16 million years after Allende calcium-aluminum-rich inclusions, during hydrothermal activity on a geologically active asteroid after chondritic components had ceased forming in the solar nebula.

Chromium Isotopes↗

Carbonates in fractures of Martian meteorite Allan Hills 84001: petrologic evidence for impact origin.

Carbonates in Martian meteorite Allan Hills 84001 occur as grains on pyroxene grain boundaries, in crushed zones, and as disks, veins, and irregularly shaped grains in healed pyroxene fractures. Some carbonate disks have tapered Mg-rich edges and are accompanied by smaller, thinner and relatively homogeneous, magnesite microdisks. Except for the microdisks, all types of carbonate grains show the same unique chemical zoning pattern on MgCO3-FeCO3-CaCO3 plots. This chemical characteristic and the close spatial association of diverse carbonate types show that all carbonates formed by a similar process. The heterogeneous distribution of carbonates in fractures, tapered shapes of some disks, and the localized occurrence of Mg-rich microdisks appear to be incompatible with growth from an externally derived CO2-rich fluid that changed in composition over time. These features suggest instead that the fractures were closed as carbonates grew from an internally derived fluid and that the microdisks formed from a residual Mg-rich fluid that was squeezed along fractures. Carbonate in pyroxene fractures is most abundant near grains of plagioclase glass that are located on pyroxene grain boundaries and commonly contain major or minor amounts of carbonate. We infer that carbonates in fractures formed from grain boundary carbonates associated with plagiociase that were melted by impact and dispersed into the surrounding fractured pyroxene. Carbonates in fractures, which include those studied by McKay et al. (1996), could not have formed at low temperatures and preserved mineralogical evidence for Martian organisms.

Calcium Carbonate↗

Petrological evidence for shock melting of carbonates in the martian meteorite ALH84001.

The meteorite ALH84001--a shocked igneous rock of probable martian origin-contains chemically and isotopically heterogeneous carbonate globules, associated with which are organic and inorganic structures that have been interpreted as possible fossil remains of ancient martian biota. A critical assumption underlying this suggestion is that the carbonates formed from low-temperature fluids penetrating the cracks and voids of the host rock. Here we report petrological studies of ALH84001 which investigate the effects of shock on the various mineralogical components of the rock. We find that carbonate, plagioclase and silica were melted and partly redistributed by the same shock event responsible for the intense local crushing of pyroxene in the meteorite. Texture and compositional data show that, during the period of shock decompression, monomineralic melts were injected into pyroxene fractures that were subsequently cooled and resealed within seconds. Our results therefore suggest that the carbonates in ALH84001 could not have formed at low temperatures, but instead crystallized from shock-melted material; this conclusion weakens significantly the arguments that these carbonates could host the fossilized remnants of biogenic activity.

Aluminum Silicates↗

Visualization of iontophoretic transport paths in cultured and animal skin models.

Physiological structures associated with iontophoretic paths in hairless mouse skin and two cultured skin models ("EpiDerm" by Mattek, Corp., and "SKIN2" by Advanced Tissue Sciences, Inc.) are reported. Visualization of ionic paths at current densities between 20 and 100 microA/ cm2 is accomplished by the counterdirectional transport of Fe(CN)6(4-) and Fe3+, resulting in the controlled precipitation of colloidal Prussian blue, Fe4[Fe(CN)6]3, at sites of high ionic conductivity. Examination of the Fe4[Fe(CN)6]3-stained tissues using optical microscopy allows unequivocal assignment of iontophoretic paths to physiological structures in the stratum corneum. Deposition of Fe4[Fe(CN)6]3 occurs exclusively at hair follicles in hairless mouse skin, indicating that these appendages provide highly conductive porous paths during iontophoresis. In contrast, the counterdirectional transport of Fe(CN)6(4-) and Fe3+ across cultured skin models, which lack appendages, results in the deposition of Fe4-[Fe(CN)6]3 along the boundaries of corneocytes. This observation suggests that paracellular iontophoretic transport through lipid bilayer regions is the predominant transport path in the absence of low-resistance pores.

Animals↗

Direct imaging of molecular transport through skin.

Scanning electrochemical microscopy (SECM) was used to image spatial variations in the molecular flux of Fe(CN)6(-4) across excised hairless and nude mouse skin. The SECM response is specific to electroactive molecules, allowing selective imaging of the flux of Fe(CN)6(-4) in multicomponent ionic solutions. Quantitative SECM image analysis demonstrated that 40% to 60% of the total Fe(CN)6(-4) flux occurred through appendages in the skin. SECM analysis of skin samples exposed to a known transport enhancer, sodium dodecylsulfate, indicated that the increase in the ion transport rate occurred exclusively in nonporous skin tissue.

Administration, Cutaneous↗

Iontophoretic transport through porous membranes using scanning electrochemical microscopy: application to in vitro studies of ion fluxes through skin.

Scanning electrochemical microscopy (SECM) is used to map localized iontophoretic fluxes of electroactive species through porous membranes. A method is described that allows both the rate of transport of species from a microscopic pore and the pore's diameter to be measured. SECM images and analyses of synthetic porous membranes (track-etched polycarbonate and mica membranes) and hairless mouse skin are reported. Preliminary analysis of SECM images of the mouse skin indicates that a significant percentage of the iontophoretic flux occurs through pores associated with hair follicles.

Aluminum Silicates↗

Transport of ionic species in skin: contribution of pores to the overall skin conductance.

Two methods are reported that allow visualization of high conductance paths in skin at current densities typically used during clinical iontophoretic drug delivery (10-200 microA/cm2). In the first method, the counter-directional iontophoretic transport of Fe(CN)6(4-) and Fe3+ across skin results in the precipitation of colloidal prussian blue, Fe4[Fe(CN)6]3, at sites of high iontophoretic flux. The appearance of localized deposits of Fe4[Fe(CN)6]3 is recorded by video microscopy and used to document the activation of low-resistance paths. In the second method, the ionic flux of Fe(CN)6(4-) through pores is directly imaged by scanning electrochemical microscopy (SECM). Both methods demonstrate that the iontophoretic flux across skin is highly localized. Activation of low-resistance pores in hairless mouse skin is shown to occur during iontophoresis. The spatial density of current carrying pores increases from 0 to 100-600 pores/cm2 during the first 30-60 min of iontophoresis. At longer times, the active pore density approaches a quasi-steady-state value that is proportional to the applied current density. The total conductance of the skin is proportional to the number of pores, consistent with a model of conduction in skin that is comprised of low-resistivity pores in parallel with a high-resistivity bulk phase. The contribution of pores to the total skin conductance during iontophoresis increases from an initial value of 0-5% to a quasi-steady-state value of 50-95%.

Animals↗

Comparison of lymphocyte immunophenotypes obtained simultaneously from two different data acquisition and analysis systems on the same flow cytometer.

Immunophenotyping of different lymphocyte populations was carried out in parallel on 113 consecutively received specimens of human peripheral blood using 2 different data acquisition and analysis systems (EPICS C and 4Cyte-Acmecyte) on the same flow cytometer (EPICS C). The phenotypes analyzed were CD3+, CD4+, CD8+ CD56+ CD16+ CD3-, TCR-gamma delta+ CD8-, and TCR-gamma delta+ CD8+. Both HIV- and HIV+ specimens were used for this study, including some with CD4 levels as low as 2% of all lymphocytes. Despite differences in gating procedures and shapes of bitmap (rectilinear vs. "amorphous"), the 2 methods agreed to within 2% positive cells in 97% of the cases. Although some statistically significant biases in the methods were observed, these were small and not biologically important. We conclude that both methods of data acquisition and analysis, as employed by experienced operators on the EPICS C flow cytometer, gave essentially equivalent results for lymphocyte sub-populations in peripheral blood preparations.

Electronic Data Processing↗

Flow cytometric analysis of gamma delta T cells and natural killer cells in HIV-1 infection.

We have previously shown that HIV-1 seropositivity is associated with an increase in the difference between the number of CD3+ lymphocytes and the total number of CD4+ and CD8+ lymphocytes [CD3 - (CD4 + CD8)] among peripheral blood lymphocytes (PBL). To investigate the cellular basis of this increase, PBL from seronegative (SN) and AIDS-free seropositive (SP) homosexual men and intravenous drug users were analyzed by two-color flow cytometry. Results showed that SP compared to SN manifested the expected elevation in calculated [CD3 - (CD4 + CD8)] cells (87 vs 28 cells/mm3; P less than 0.001). Only small differences in lymphocyte populations that could contribute to this increase were observed, namely lymphocytes expressing the CD3+CD4-CD8-phenotype (67 vs 56 cells/mm3; P greater than 0.10) or the CD8dim phenotype (135 vs 142 cells/mm3; P greater than 0.10). However, SP had significantly lower numbers of cells expressing the CD56+CD3- phenotype characteristic of natural killer cells (81 vs 170 cells/mm3; P less than 0.001) and significantly higher numbers of T cells expressing the gamma delta T cell receptor (TCR) (81 vs 62 cells/mm3; P = 0.10). The latter difference was primarily due to higher numbers of cells coexpressing gamma delta-TCR and low levels of CD8 (27 vs 15 cells/mm3; P = 0.009). These data suggest that HIV-1 seropositivity is associated with low numbers of natural killer cells and high numbers of CD8+ gamma delta-TCR lymphocytes. Changes in these populations may reflect altered host defense against HIV-1 or altered T cell kinetics in the presence of HIV-1 infection.

Acquired Immunodeficiency Syndrome↗

Flow cytometric quantitation of T cell phenotypes in cerebrospinal fluid and peripheral blood of homosexual men with and without antibodies to human immunodeficiency virus, type I.

Two-color flow cytometry was used to analyze T cell subsets (total (CD3), helper-inducer (CD4), and suppressor-cytotoxic (CD8] in paired specimens of cerebrospinal fluid (CSF) and peripheral blood of 66 homosexual men, including 62 with antibodies to human immunodeficiency virus, type 1 (HIV-1). With the exception of one traumatic specimen, all of the CSF specimens, 52 of which had less than or equal to 5 lymphocytes/mm3, were evaluated fully, with the number of lymphocytes counted for each antibody ranging from 200 to 2933 (mean = 1129). Proportions of CD3, CD4, and CD8 lymphocytes in CSF were very highly correlated with the proportions of these cells in the peripheral blood (r = 0.87, 0.96, and 0.94, respectively), as was the CD4/CD8 ratio (r = 0.98). These strong correlations were present in each of seven subgroups of study subjects defined on the basis of detailed neurologic examination, neuropsychological testing, and the presence or absence of antibodies to HIV-1. In the population studied, T cell phenotypes in CSF as analyzed by two-color flow cytometry were largely determined by the corresponding proportions in the peripheral blood.

Adult↗