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Biomedical subjects

A Schaper

Publications and source records attributed to A Schaper.

31 records · Page 2Linked to original sources
PubMed1994

Probing chromatin with the scanning force microscope.

With the scanning force microscope (SFM), one can image the topography of biological material adsorbed at air-solid or liquid-solid interfaces with up to nanometer resolution. In principle, fixation, contrast enhancement, and labeling are not required. We have adapted specimen preparation techniques of conventional electron microscopy for visualizing chromatin ultrastructures in the SFM. A beaded substructure of the nucleoprotein filament was obtained after hypotonic lysis of chicken erythrocytes and air drying. The beads-on-a-string morphology of the basic nucleosomal assembly was well delineated. The nucleosomes appeared as round protrusions with an apparent height of 4-6 nm. The histogram of center-to-center distances between adjacent nucleosome cores along the filament axis had a peak at approximately 30 nm. Reversible changes in the three-dimensional structure were observed upon exposure of air-dried samples of metaphase chromosomes to solutions of different ionic strengths.

Animals↗
PubMed1994

Imaging subcellular structures of rat mammary carcinoma cells by scanning force microscopy.

Scanning force microscopy (SFM) was used for imaging subcellular structures of cultured rat mammary carcinoma cells dried in air. Identification of cellular substructures was achieved by immunofluorescence and specific fluorescence probes. Cells grown attached to a glass support exhibited submicrometer thickness in the dried state. Inside the nuclear domain the nucleoli appeared as prominent conical protrusions. Membrane extensions, microspikes and microvilli were well preserved at the cell periphery after fixation in glutaraldehyde vapor and air-drying and were distinguishable either as isolated elements or intercellular communications. The plasma membrane and soluble proteins were selectively removed with nonionic detergent in a buffer system. The mitochondria were concentrated primarily in the perinuclear space and exhibited a well defined filamentous shape. Their identity was confirmed by specific fluorescence staining with rhodamine 123. In the membrane-free system achieved by dry-cleaving of the sample surface, the cytoskeletal network was resolved as a complex mesh of actin-containing fiber bundles interwoven with a filigree arrangement of thinner filaments. The smallest fibrous substructures revealed by SFM with the scanning tips used to date were approximately 8 to 10 nm in height and 80 nm in width.

Animals↗
PubMed1994

Dysfunctional labor after external cephalic version.

OBJECTIVE: To determine whether there is a greater occurrence of dysfunctional labor or a higher incidence of cesarean delivery for failure to progress in women who have undergone a successful version for breech presentation. METHODS: Using a retrospective case-control design, 76 women who had undergone a successful version from January 1988 through July 1993 were identified and their medical records reviewed. The control population was matched for delivery date, parity, and gestational age. RESULTS: There was no difference in the cesarean delivery rates for failure to progress between women undergoing successful version (6%) and the control population (6%). The incidence of dysfunctional labor between the groups did not differ significantly. Dysfunctional labor requiring oxytocin augmentation occurred in 29.6% of women who had undergone a version and in 24.2% of the control population (P = .6530). CONCLUSION: There is no significant increase in the cesarean delivery rate in women undergoing a version for breech presentation.

Case-Control Studies↗
PubMed1993

Scanning force microscopy of circular and linear plasmid DNA spread on mica with a quaternary ammonium salt.

Scanning force microscopy (SFM) offers the potential for subnanometer resolution in the investigation of nucleic acids, proteins, and their complexes. SFM is not bound by the requirement of classical transmission electron microscopy (TEM) for contrast enhancement through shadow casting or negative staining. A primary challenge, however, has been the reproducible fixation of samples on an atomically flat surface such as mica. We have developed a method for the routine imaging by SFM of supercoiled, relaxed, and linearized plasmid DNA, immobilized on freshly cleaved mica through the spreading action of benzyldimethylalkylammonium chloride (BAC) at micromolar concentrations. A reproducibly high yield of well-spread, dispersed molecules is obtained and background contamination is minimal. The contour lengths of the relaxed and linearized molecules imaged in air agree well with the helical rise (3.4 A/bp) of B-DNA in solution. We have also introduced the use of quantitative image analysis of SFM images to determine apparent molecular width and height over the entire molecular path.

DNA↗
PubMed1991

Salt dependent changes in structure and dynamics of circular single stranded DNA of filamentous phages of Escherichia coli.

We have analyzed the static and dynamic behaviour of the circular single stranded DNA of the filamentous Escherichia coli phages F1 and M13mp8 in solution as a function of salt concentration using static and dynamic light scattering and sedimentation analysis in the analytical ultracentrifuge. We show by static light scattering that native and denatured single stranded DNA behave like a randomly coiled macromolecule at all salt concentrations used. The size of the native single stranded DNA is governed by the formation of secondary structures. While the radius of gyration decreases with increasing salt concentration the translational diffusion of the center-of-mass of native single stranded DNA and the sedimentation coefficient increase with increasing salt concentration in a biphasic manner. Below 100 mM monovalent cation concentration there is a strong dependence of the hydrodynamic parameters upon salt which is reduced approx. 3-fold at higher salt concentrations. We attribute the compaction of single stranded DNA by salt to electrostatic shielding and, in case of native single stranded DNA, secondary structure formation. Internal motions of the native single stranded DNA are observable at all salt concentrations and can be interpreted with a model of segmental diffusion of the elements of the polymer chain. The observed segmental diffusion coefficient of the native single stranded polynucleotide increases with increasing salt under the conditions investigated.

Coliphages↗
PubMed1991

Structure and dynamics of the complex of single stranded DNA binding protein of Escherichia coli with circular single stranded DNA of filamentous phages.

We have analyzed the equilibrium and nonequilibrium properties of the complex of the single stranded DNA binding protein of Escherichia coli (EcoSSB) and circular single stranded DNA of filamentous phages M13mp8 and F1 using static and dynamic light scattering, analytical ultracentrifugation and electron microscopy. Upon binding to the single stranded DNA the EcoSSB tetramer replaces an equivalent volume of water trapped within the coiled single stranded DNA and hinders the folding of the single stranded DNA into secondary structures at all salt concentrations. The salt dependent compaction of the stoichiometric complex can be described assuming a flexible polyelectrolyte chain. The solution structure of the macromolecular complex is a random coil and in the electron microscope a beaded flexible structure of the complex with a bead diameter of 6 nm appears at all salt concentrations used. The internal motions of the stoichiometric complex can be described by the Rouse-Zimm model of polymer dynamics. The segmental mobility of the complex can be correlated with changes in the binding site size of the EcoSSB tetramer; it indicates the presence of interactions between EcoSSB tetramers bound to single stranded DNA.

Coliphages↗
PubMed1991

[Disorders of wound healing of the sacral cavity after abdomino-perineal resection of the rectum].

In 95 patients it was necessary to perform abdominoperineal resection and colostomy due to malignant tumours of the rectum, the anus, as well as gynaecological tumours. After standardized preoperative preparation and comparable operative techniques in more than 93 per cent of the cases we performed suction drainage of the sacral cavity. In 86 per cent we observed primary perineal wound closure and the patients were spared lengthy secondary wound healing.

Adult↗
PubMed1990

Kinetics of binding of single-stranded DNA binding protein from Escherichia coli to single-stranded nuclei acids.

The time course of the reaction of Escherichia coli single-stranded DNA binding protein (E. coli SSB) with poly(dT) and M13mp8 single-stranded DNA has been measured by fluorescence stopped-flow experiments. For poly(dT), the fluorescence traces follow simple bimolecular behavior up to 80% saturation of the polymer with E. coli SSB. A mechanistic explanation of this binding behavior can be given as follows: (1) E. coli SSB is able to translocate very rapidly on the polymer, forming cooperative clusters. (2) In the rate-limiting step of the association reaction, E. coli SSB is bound to the polymer only by one or two of its four contact sites. As compared to poly(dT), association to single-stranded M13mp8 phage DNA is slower by at least 2 orders of magnitude. We attribute this finding to the presence of secondary structure elements (double-stranded structures) in the natural single-stranded DNA. These structures cannot be broken by E. coli SSB in a fast reaction. In order to fulfill its physiological function in reasonable time, E. coli SSB must bind newly formed single-stranded DNA immediately. The protein can, however, bind to such pieces of the newly formed single-stranded DNA which are too short to cover all four binding sites of the E. coli SSB tetramer.

Coliphages↗
PubMed1989

Acute renal failure in octogenarians.

We analyzed hospital survival rates, severity of illness, and long-term outcomes in patients from three different age groups who were treated with hemodialysis for acute renal failure (ARF). Patients over age 80 had fewer bad prognostic factors. This selection bias was reflected in their hospital survival rates of 53%, compared to 57% of patients under age 70. No selection bias for patients ages 70 to 79 years was identified, and their hospital survival rate was 27%. The long-term outcome of hospital survivors was poor, particularly in the very elderly. Physicians should consider all of these factors when deciding whether to offer hemodialysis to elderly patients suffering from ARF.

Acute Kidney Injury↗
PubMedDate not supplied

Scanning force microscopy of chromatin fibers in air and in liquid.

We have adapted specimen preparation techniques of conventional electron microscopy for visualizing chromatin structures in the scanning force microscope (SFM) in air and in liquid. The beaded substructure of the nucleoprotein filament was obtained after hypotonic lysis of chicken erythrocytes and air drying, whereas supranucleosomal structures were preserved after treatment of cell nuclei with detergent. In the latter case, the nucleosomes were still distinct but appeared more condensed. A modified droplet diffusion-spreading technique of chromatin from Namalwa cells (a human B-lymphoid line) yielded a uniform filamentous morphology and similar fiber appearance. A reversible swelling of spread chromatin was observed upon exposure of air-dried samples to solutions differing in salt concentrations.

Animals↗
PubMedDate not supplied

Sterilization of contrast media (Isovist) containing liposomes by ethylene oxide.

Liposomes containing Isovist were prepared by controlled detergent dialysis as well as by reverse phase evaporation. After preparation these liposomes were lyophilized and then submitted to sterilization by ethylene oxide. Prior to lyophilization, trehalose was added as a protective agent to preserve the size of the liposomes. After each step in the preparation size distribution was determined by photon correlation spectroscopy. The computer program CONTIN was adapted for the data analysis and proved to be applicable for polydisperse solutions of liposomes. Neither freeze-drying nor sterilization had negative effects on the morphological quality of the samples when using 4 g of trehalose per 1 g of lipid. In addition, the quality of the liposomes was controlled by scanning electron microscopy. At the end of seven days, no growth of microorganisms occurred in any of the samples.

Contrast Media↗