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B Drake

Publications and source records attributed to B Drake.

35 records · Page 2Linked to original sources

Immobilized proteins in buffer imaged at molecular resolution by atomic force microscopy.

Samples of supported planar lipid-protein membranes and actin filaments on mica were imaged by atomic force microscopy (AFM). The samples were fully submerged in buffer at room temperature during imaging. Individual proteins bound to the reconstituted membrane were distinguishable; some structural details could be resolved. Also, surface-induced, self-assembling of actin filaments on mica could be observed. Monomeric subunits were imaged on individual actin filaments. The filaments could be manipulated on or removed from the surface by the tip of the AFM. The process of the decoupling of the filamentous network from the surface upon changing the ionic conditions was imaged in real time.

Actins↗

Imaging crystals, polymers, and processes in water with the atomic force microscope.

The atomic force microscope (AFM) can be used to image the surface of both conductors and nonconductors even if they are covered with water or aqueous solutions. An AFM was used that combines microfabricated cantilevers with a previously described optical lever system to monitor deflection. Images of mica demonstrate that atomic resolution is possible on rigid materials, thus opening the possibility of atomic-scale corrosion experiments on nonconductors. Images of polyalanine, an amino acid polymer, show the potential of the AFM for revealing the structure of molecules important in biology and medicine. Finally, a series of ten images of the polymerization of fibrin, the basic component of blood clots, illustrate the potential of the AFM for revealing subtle details of biological processes as they occur in real time.

Crystallography↗

The scanning ion-conductance microscope.

A scanning ion-conductance microscope (SICM) has been developed that can image the topography of nonconducting surfaces that are covered with electrolytes. The probe of the SICM is an electrolyte-filled micropipette. The flow of ions through the opening of the pipette is blocked at short distances between the probe and the surface, thus, limiting the ion conductance. A feedback mechanism can be used to maintain a given conductance and in turn determine the distance to the surface. The SICM can also sample and image the local ion currents above the surfaces. To illustrate its potential for imaging ion currents through channels in membranes, a topographic image of a membrane filter with 0.80-micrometer pores and an image of the ion currents flowing through such pores are presented.

Cell Membrane↗

STM and AFM images of nucleosome DNA under water.

We have imaged DNA from the calf thymus nucleosome using a scanning tunneling microscope (STM) operated in water. The fragments are deposited onto the interface between a buffer solution and an epitaxially grown gold surface using an electrochemical tecnique. Most of the fragments are fairly straight, and when individual polymers can be identified, their length is consistent with the expected 146 basepairs (approximately 500 A). The resolution is often adequate to show signs of the 36 A helical pitch. Some images show a structure which appears to have abrupt kinks of the sort predicted by Crick and Klug (Nature 255, 530-533, 1975). In order to check that this shape is not a consequence of binding to underlying structure on the gold substrate, we have also made images of kinked structures using an atomic force microscope (AFM) with the DNA bound to glass.

Animals↗

Atomic force microscopy of an organic monolayer.

Atomic force microscope images of polymerized monolayers of n-(2-aminoethyl)-10,12-tricosadiynamide revealed parallel rows of molecules with a side-by-side spacing of approximately equal to 0.5 nanometer. Forces used for imaging (10(-8) newton) had no observable effect on the polymer strands. These results demonstrate that atomic force microscope images can be obtained for an organic system.

Amides↗

Scanning probe microscopy of biological samples and other surfaces.

Scanning probe microscopes derived from the scanning tunnelling microscope (STM) offer new ways to examine surfaces of biological samples and technologically important materials. The surfaces of conductive and semiconductive samples can readily be imaged with the STM. Unfortunately, most surfaces are not conductive. Three alternative approaches were used in our laboratory to image such surfaces. 1. Crystals of an amino acid were imaged with the atomic force microscope (AFM) to molecular resolution with a force of order 10(-8) N. However, it appears that for most biological systems to be imaged, the atomic force microscope should be able to operate at forces at least one and perhaps several orders of magnitude smaller. The substitution of optical detection of the cantilever bending for the measurement by electron tunnelling improved the reliability of the instrument considerably. 2. Conductive replicas of non-conductive surfaces enabled the imaging of biological surfaces with an STM with a lateral resolution comparable to that of the transmission electron microscope. Unlike the transmission electron microscope, the STM also measures the heights of the features. 3. The scanning ion conductance microscope scans a micropipette with an opening diameter of 0.04-0.1 micron at constant ionic conductance over a surface covered with a conducting solution (e.g., the surface of plant leaves in saline solution).

Crystallization↗

The immunochemical characterisation of circulating immune complex constituents in Candida albicans osteomyelitis by isoelectric focusing, immunoblot, and immunoprint.

Circulating immune complexes present in the serum of a patient with systemic lupus erythematosus, end-stage renal failure, and thoracic vertebral Candida albicans osteomyelitis were sequentially analysed by isoelectric focusing, immunoblotting, and immunoprinting. Candida antigens (including mannoproteins), Clq and C3 complement components, and specific anti-Candida antibody were detected within polyethylene glycol precipitated complexes. An antigen of 47K molecular weight was amongst those demonstrated by polyacrylamide gel electrophoresis to be present within the complexes. It has been proposed elsewhere that a serologic response to a 47K protein is predictive of recovery from Candida albicans infection. Free antibody had specificity for Candida antigens ranging in molecular weight from 18K to more than 100K including a 47K component. The analytical techniques employed allowed rapid and precise identification of the components of one particular immune complex system and will be widely applicable to the dissection of other diverse systems.

Adult↗

Determination of cerebral death in the pediatric intensive care unit.

The clinical course, laboratory data, outcome, and autopsy findings in 61 pediatric patients with suspected brain death were reviewed. In 58% of patients, the initial EEG was isoelectric, and cerebral blood flow was absent. In six of nine children, the initial EEG, which showed activity, became isoelectric by 72 hours. In no child without demonstrable cerebral flow did flow resume in later studies. In four of five children who had initial EEG activity despite absent cerebral flow, an isoelectric EEG developed on repeated study. The average time from initial insult until clinically suspected brain death was 29.5 hours and 61.5 hours until brain death was confirmed. The time from confirmation of brain death until discontinuation of life support systems was 32 hours in the majority of patients. Our current protocol for evaluating pediatric patients with suspected brain death is reviewed with emphasis on the clinical examination, laboratory studies, and use of serial EEGs and radionuclide cerebral blood flow determinations.

Adolescent↗

Purification and properties of glutathione S-transferases from larvae of Wiseana cervinata.

The glutathione S-transferases from the porina moth, Wiseanna cervinata, were purified by affinity chromatography, cation-exchange chromatography and preparative isoelectrofocusing. The major transferase (IV) was purified to homogeneity by a factor of 530-fold with a yield of 83%. Other transferases present were purified to a smaller degree (approx. 50-fold) to a stage of near-homogeneity. The transferases examined all had Mr values about 45 000-50 000. They appeared to be homodimers of either of two types of subunit, of Mr 22 800 and 24 600. Enzymes consisting of the different types of subunit were not immunologically cross-reactive. The major enzyme fractions separated by cation-exchange chromatography were both active with 1-chloro-2,4-dinitrobenzene, 1,2-dichloro-4-nitrobenzene, ethacrynic acid and iodomethane, but were inactive with 4-nitropyridine N-oxide, 1,2-epoxy-3-(p-nitrophenoxy)propane, bromosulphophthalein and p-nitrobenzyl chloride. The kinetics of the enzyme-catalysed reaction with enzyme IV were non-Michaelean with respect to both substrates. Both products were inhibitory. The results appear to be compatible with a random steady-state mechanism. It is concluded that these enzymes are very similar, in their physical and chemical constitution, in their catalytic properties and in their relationships with each other, to those enzymes that have been isolated from vertebrate organisms.

Amino Acids↗

Prechilling of Xanthium strumarium L. Reduces Net Photosynthesis and, Independently, Stomatal Conductance, While Sensitizing the Stomata to CO(2).

Greenhouse-grown plants of Xanthium strumarium L. were exposed in a growth cabinet to 10 C during days and 5 C during nights for periods of up to 120 hours. Subsequently, CO(2) exchange, transpiration, and leaf temperature were measured on attached leaves and in leaf sections at 25 or 30 C, 19 C dew point of the air, 61 milliwatts per square centimeter irradiance, and CO(2) concentrations between 0 and 1000 microliters per liter ambient air. Net photosynthesis and stomatal conductance decreased and dark respiration increased with increasing duration of prechilling. The reduction in net photosynthesis was not a consequence of decreased stomatal conductance because the intercellular CO(2) concentration in prechilled leaves was equal to or greater than that in greenhouse-grown controls. The intercellular CO(2) concentration at which one-half maximum net photosynthesis occurred remained the same in prechilled leaves and controls (175 to 190 microliters per liter). Stomata of the control plants responded to changes in the CO(2) concentration of the air only slightly. Prechilling for 24 hours or more sensitized stomata to CO(2); they responded to changes in CO(2) concentration in the range from 100 to 1000 microliters per liter.

Journal Article↗

Effects of elevated carbon dioxide on soils in a Florida scrub oak ecosystem.

The results of a 3-yr study on the effects of elevated CO2 on soil N and P, soil pCO2, and calculated CO2 efflux in a fire-regenerated Florida scrub oak ecosystem are summarized. We hypothesized that elevated CO2 would cause (i) increases in soil pCO2 and soil respiration and (ii) reduced levels of soil-available N and P. The effects of elevated CO2 on soil N availability differed according to the method used. Results of resin lysimeter collections and anion exchange membrane tests in the field showed reduced NO3- in soils in Years 1 and 3. On the other hand, re-analysis of homogenized, buried soil bags after 1 yr suggested a relative increase in N availability (lower C to N ratio) under elevated CO2. In the case of P, the buried bags and membranes suggested a negative effect of CO2 on P during the first year; this faded over time, however, as P availability declined overall, probably in response to P uptake. Elevated CO2 had no effect on soil pCO2 or calculated soil respiration at any time, further suggesting that plant rather than microbial uptake was the primary factor responsible for the observed changes in N and P availability with elevated CO2.

Biological Availability↗