Biomedical subjects

A Engel

Publications and source records attributed to A Engel.

At least 91 records · Page 5Linked to original sources

PubMed1999

Voltage and pH-induced channel closure of porin OmpF visualized by atomic force microscopy.

Gram-negative bacteria are protected by an outer membrane in which trimeric channels, the porins, facilitate the passage of small solutes. The pores are formed by membrane-spanning antiparallel beta-strands, which are connected by short turns on the periplasmic side and long loops on the extracellular side. Voltage and pH-dependent conformational changes of these extracellular loops have now been visualized by atomic force microscopy of two-dimensional crystals of Escherichia coli porin OmpF. The observed conformational changes accompany the closure of the channel entrance, and suggest that this is a mechanism that the cells have evolved to protect themselves from drastic changes of the environment.

Bacterial Proteins↗

PubMed1999

Organising multi-dimensional biological image information: the BioImage Database.

Nowadays it is possible to unravel complex information at all levels of cellular organization by obtaining multi-dimensional image information. At the macromolecular level, three-dimensional (3D) electron microscopy, together with other techniques, is able to reach resolutions at the nanometer or subnanometer level. The information is delivered in the form of 3D volumes containing samples of a given function, for example, the electron density distribution within a given macromolecule. The same situation happens at the cellular level with the new forms of light microscopy, particularly confocal microscopy, all of which produce biological 3D volume information. Furthermore, it is possible to record sequences of images over time (videos), as well as sequences of volumes, bringing key information on the dynamics of living biological systems. It is in this context that work on BioImage started two years ago, and that its first version is now presented here. In essence, BioImage is a database specifically designed to contain multi-dimensional images, perform queries and interactively work with the resulting multi-dimensional information on the World Wide Web, as well as accomplish the required cross-database links. Two sister home pages of BioImage can be accessed at http://www. bioimage.org and http://www-embl.bioimage.org

Bacterial Proteins↗

PubMed1999

Two-dimensional crystallization of Escherichia coli lactose permease.

A chimeric protein consisting of lactose permease with cytochrome b562 in the middle cytoplasmic loop and six His residues at the C terminus (LacY/L6cytb562/417H6 or "red permease") was overexpressed in Escherichia coli and isolated by nickel affinity chromatography after solubilization with dodecyl-beta,d-maltopyranoside. Red permease was then reconstituted in the presence of phospholipids, yielding densely packed vesicles and well-ordered two-dimensional (2D) crystals as shown by electron microscopy of negatively stained specimens. Single-particle analysis of 16 383 protein particles in densely packed vesicles reveals a 5.4-nm-long trapeziform protein of 4.1 to 5.1 nm width, with a central stain-filled indentation. Depending on reconstitution conditions, trigonal and rectangular crystallographic packing arrangements of these elongated particles assembled into trimers are observed. The best ordered 2D crystals exhibit a rectangular unit cell, of dimensions a = 9.9 nm, b = 17.4 nm, that houses two trimeric complexes. Projection maps calculated to a resolution of 2 nm show that these crystals consist of two layers.

Carrier Proteins↗

PubMed1999

Editorial

Explore the source record for details and available documents.

Journal Article↗

PubMed1999

Prerequisites for the immunotherapy of cancer.

Tumours express proteins not commonly found in normal cells, or over-express certain proteins. These may in some cases serve as target antigens for immunological attack. It is therefore essential to improve our understanding of the nature of these target epitopes and the cells which recognize them, in order to develop immunotherapy as a realistic treatment for cancer. A small group of around 40 investigators recently came together at the Heinrich Fabri Institute of the University of Tübingen to discuss the identification of human tumour antigens and the exploitation of this knowledge for effective immunotherapy.

Antigen-Presenting Cells↗

PubMed1999

Tapping-mode atomic force microscopy produces faithful high-resolution images of protein surfaces.

Compared to contact-mode atomic force microscopy (CMAFM), tapping-mode atomic force microscopy (TMAFM) has the advantage of allowing imaging surfaces of macromolecules, even when they are only weakly attached to the support. In this study, TMAFM is applied to two different regular protein layers whose structures are known to great detail, the purple membrane from Halobacterium salinarum and the hexagonally packed intermediate (HPI) layer from Deinococcus radiodurans, to assess the faithfulness of high-resolution TMAFM images. Topographs exhibited a lateral resolution between 1.1 and 1. 5 nm and a vertical resolution of approximately 0.1 nm. For all protein surfaces, TMAFM and CMAFM topographs were in excellent agreement. TMAFM was capable of imaging the fragile polypeptide loop connecting the transmembrane alpha-helices E and F of bacteriorhodopsin in its native extended conformation. The standard deviation (SD) of averages calculated from TMAFM topographs exhibited an enhanced minimum (between 0.1 and 0.9 nm) that can be assigned to the higher noise of the raw data. However, the SD difference, indicating the flexibility of protein subunits, exhibited an excellent agreement between the two imaging modes. This demonstrates that the recently invented imaging-mode TMAFM has the ability to faithfully record high-resolution images and has sufficient sensitivity to contour individual peptide loops without detectable deformations.

Bacterial Proteins↗

PubMed1999

Electrostatically balanced subnanometer imaging of biological specimens by atomic force microscope.

To achieve high-resolution topographs of native biological macromolecules in aqueous solution with the atomic force microscope (AFM) interactions between AFM tip and sample need to be considered. Short-range forces produce the submolecular information of high-resolution topographs. In contrast, no significant high-resolution information is provided by the long-range electrostatic double-layer force. However, this force can be adjusted by pH and electrolytes to distribute the force applied to the AFM tip over a large sample area. As demonstrated on fragile biological samples, adjustment of the electrolyte solution results in a local reduction of both vertical and lateral forces between the AFM tip and proteinous substructures. Under such electrostatically balanced conditions, the deformation of the native protein is minimized and the sample surface can be reproducibly contoured at a lateral resolution of 0.6 nm.

Bacterial Proteins↗

PubMed1999

T cell immunosenescence in vitro and in vivo.

The term "immunosenescence" refers to an age-associated dysregulation of immune function which contributes to the increased susceptibility of the elderly to infectious disease. Although there are age-associated changes measurable in the innate immune system (Pawelec et al., 1998c), it is the adaptive, particularly T cell, system which is most susceptible to the deleterious effects of aging. In this minireview, characteristics of aging in long-term human T cell cultures will be summarized, and the parallels between the in vitro model and in vivo immunosenescence will be documented. The use of culture models to screen for ways of manipulating immunosenescence in vitro may provide a basis for intervention to ameliorate immunosenescence in vivo.

Aging↗

PubMed1999

Atomic force microscopy: a powerful tool to observe biomolecules at work.

Atomic force microscopes (AFMs) move a sharp tip attached to a soft cantilever in a TV-raster-like pattern over a surface and record deflections of the tip that correspond to the surface topography. When operated in physiological solutions, an AFM allows biomolecules to be observed in their native environment. Progress in instrumentation, sample-preparation methods and recording conditions has provided images of biomolecules and their assemblies that reveal submolecular details. In addition, the AFM allows conformational changes to be observed directly. This article discusses these points and illustrates them with some pertinent examples.

Microscopy, Atomic Force↗

PubMed1999

Imaging streptavidin 2D crystals on biotinylated lipid monolayers at high resolution with the atomic force microscope.

Streptavidin crystals were grown on biotinylated lipid monolayers at an air/water interface and transferred onto highly oriented pyrolytic graphite (HOPG). These arrays could be imaged to a resolution below 1 nm using the atomic force microscope. The surface topographs obtained were compared with negative-stain electron microscopy images and the atomic model as determined by X-ray crystallography. The streptavidin tetramer (60 kDa) exposes two free biotin-binding sites to the buffer solution, while two are occupied by linkage to the lipid monolayer. Therefore, the streptavidin 2D crystals can be used as nanoscale matrices for binding biotinylated compounds. Furthermore, this HOPG-based preparation method provides a general novel approach to study the structure of protein arrays assembled on lipid monolayers with the AFM.

Journal Article↗

PubMed1999

Robotic selective sampling and total automation for anti-CMV screening.

The provision of anti-cytomegalovirus (CMV)-negative blood products causes the blood banking community considerable logistical problems. This is due firstly to the fact that only a select group of patients require anti-CMV-negative units, namely the immunocompromised. Secondly, the prevalence of CMV antibody in the blood donor population is high and, thirdly, the demand for anti-CMV-negative blood components continues to increase. To address this problem, a system of robotic selective sampling and total assay automation for anti-CMV screening has been developed. An in-house computer program generates a worklist from the donation database of samples requiring screening to meet clinical demand. Algorithms incorporated in the program ensure that the minimum number of samples possible are selected for provision of anti-CMV-negative products. This 'intelligent' selection of samples substantially reduces reagent cost. Robotic pipetting from the electronically derived worklist, masterplate to test plate robotic transfer and use of an automated assay processor maintains specific sample identification throughout. Results are read mechanically and transmitted directly to the main computer system. Robotic selective sampling and total assay automation produces an efficient, secure, auditable and completely traceable system for the provision of anti-CMV-negative units. The system produced a substantial reduction in labour costs with a 40% reduction in reagent usage. This system can be readily adapted for screening other markers on a selective basis.

Antibodies, Viral↗

PubMed1999

Investigation of the structure of spinach photosystem II reaction center complex.

The photosystem II (PSII) reaction center (RC) complex was isolated from spinach and characterized by gel electrophoresis, gel filtration and analytical ultracentrifugation. The purified complex contained the PsbA, PsbD, PsbE, PsbF and PsbI subunits. Gel filtration and analytical ultracentrifugation indicated the presence of a homogeneous complex. The mass of the RC complexes was found to be 107 kDa by analytical ultracentrifugation and 132 kDa by scanning transmission electron microscopy (STEM). The mass obtained showed the isolated complex to exist as a monomer and only one cytochrome b559 (cyt b559) to be associated with the RC complex. Digital images of negatively stained RC complexes were recorded by STEM and analyzed by single-particle averaging. The complex was 9 nm long and 5 nm wide, and exhibited a pronounced quasi-twofold symmetry. This supports the symmetric organization of the PSII complex, with the PsbA and the PsbD proteins in the center and symmetrically arranged PsbB and PsbC proteins at the periphery of the monomeric complex.

Electrophoresis, Polyacrylamide Gel↗

PubMed1999

[Years of training: a new risk factor in acute badminton injuries].

The incidence of badminton injuries is low compared with other sports, but acute injuries are generally more severe. Little is known about the risk of competitive badminton players to get an acute badminton injury. The purpose of this study was to define for the first time "years of playing badminton on a competitive level" as a risk factor for acute badminton injuries. 179 badminton injuries of 102 Austrian competitive badminton players, some of them being elite players in the european championships, were retrospectively registered. The years 1993, 1994 and 1995 were covered by our investigation. Injury incidences were defined as injuries/1000 h and were calculated separately for the 0. through the 21st year of competitive badminton. The incidence of acute badminton injuries increased constantly from the 0 to the 7th year of competitive badminton. The 6th and 7th year of competitive badminton showed a threefold increased incidence of acute badminton injuries when compared with the first year and the late years of the career as a competitive badminton player. We found no correlation between the incidence of acute badminton injuries and age, gender, hours played per year or time of warming up. We conclude that players being engaged in competitive badminton for 5 to 8 years represent a high risk population for acute badminton injuries. In contrast players at the beginning and towards the end of the competitive career showed a markedly lower risk for acute badminton injuries. We recommend additional training efforts concerning technical skills and endurance as well as proper rehabilitation of sports injuries for the high risk group of badminton players.

Adolescent↗

PubMed1999

Diagnostic value of procalcitonin serum levels in neutropenic patients with fever: comparison with interleukin-8.

We assessed the predictive value of procalcitonin (PCT) serum levels in neutropenic patients with fever and various types of infection, using a prospective 3 times weekly blood sampling protocol during 103 patient episodes. Compared with pre-fever levels, median PCT levels increased after fever onset from 0.16 ng/ml (day -1) to 0.34 ng/ml (day +1). In samples obtained within 32 h after fever onset, PCT levels differed significantly between (clinically or microbiologically) documented infection and unexplained fever (median 0.51 vs. 0.26 ng/ml), between bacteraemia and non-bacteraemic infection (median 0.8 vs. 0.27 ng/ml) and between Gram-negative bacteraemia and all other episodes (median 1.28 vs. 0.31 ng/ml). Receiver-operating-characteristic (ROC) curves indicated that the discriminatory power of PCT was best for predicting bacteraemia vs. non-bacteraemic infection (sensitivity 73%; specificity 86%; area under the ROC curve 0.795; cut-off value 0.5 ng/ml). Compared with interleukin-8 (IL-8) serum levels, test characteristics were similar in the prediction of bacteraemia vs. non-bacteraemic infection and in the prediction of documented infection vs. unexplained fever, while IL-8 was better than PCT in the prediction of Gram-negative bacteraemia (area under the ROC curve 0.965 vs. 0.758).

Adolescent↗

PubMed1999

Epithelioid angiosarcoma associated with a Dacron vascular graft.

Angiosarcoma developed at the site of a Dacron vascular prosthesis 8 years after an aortobifemoral bypass graft insertion. The tumor was composed of epithelioid cells, which showed positive staining for cytokeratin and expression of the common endothelial markers CD31, CD34, and von Willebrand factor. Ultrastructural examination showed aggregates of large cells with intercellular lumina and focal perinuclear whorls of intermediate filaments. The patient, who had abdominal pain and weight loss, died of disseminated pelvic and abdominal disease 6 months after diagnosis. Sarcomas associated with vascular Dacron grafts and angiosarcomas associated with metal or polymer foreign bodies are rare. Their development is probably analogous to the common experimental development of foreign body-associated sarcomas in rodents. Physicians caring for patients with vascular grafts or metal foreign bodies should be aware of this complication.

Aged↗

PubMed1999

Correlations between hidden units in multilayer neural networks and replica symmetry breaking.

We consider feed-forward neural networks with one hidden layer, tree architecture, and a fixed hidden-to-output Boolean function. Focusing on the saturation limit of the storage problem the influence of replica symmetry breaking on the distribution of local fields at the hidden units is investigated. These field distributions determine the probability of finding a specific activation pattern of the hidden units as well as the corresponding correlation coefficients and therefore quantify the division of labor among the hidden units. We find that although modifying the storage capacity and the distribution of local fields markedly replica symmetry breaking has only a minor effect on the correlation coefficients. Detailed numerical results are provided for the PARITY, COMMITTEE, and AND machines with K=3 hidden units and nonoverlapping receptive fields.

Journal Article↗

PubMed1999

Dissipation in ferrofluids: mesoscopic versus hydrodynamic theory.

Part of the field dependent dissipation in ferrofluids occurs due to the rotational motion of the ferromagnetic grains relative to the viscous flow of the carrier fluid. The classical theoretical description due to Shliomis (Zh. Eksp. Teor. Fiz. 61, 2411 (1971) [Sov. Phy JETP 34, 1291 (1972)]) uses a mesoscopic treatment of the particle motion to derive a relaxation equation for the nonequilibrium part of the magnetization. Complementary, the hydrodynamic approach of Liu [Phys. Rev. Lett. 70, 3580 (1993)] involves only macroscopic quantities and results in dissipative Maxwell equations for the magnetic fields in the ferrofluid. Different stress tensors and constitutive equations lead to deviating theoretical predictions in those situations, where the magnetic relaxation processes cannot be considered instantaneous on the hydrodynamic time scale. We quantify these differences for two situations of experimental relevance, namely, a resting fluid in an oscillating oblique field and the damping of parametrically excited surface waves. The possibilities of an experimental differentiation between the two theoretical approaches is discussed.

Journal Article↗

PubMed1999

Methods for estimating the sound pressure at the eardrum.

The problem of estimating the sound pressure generated at individual eardrums is systematically investigated. In audiometry, the reference to the pressure at the eardrum is usually realized by using a coupler such as the IEC 711 ear simulator, which is intended to approximate an average ear. The errors caused by individually shaped ear canals are calculated for a typical audiometric earphone (Beyer DT 48) in combination with the IEC 711 ear simulator and with an "ideal" coupler. These errors can reach 15 dB and are clearly more important than deviations of the ear simulator from an average ear. In order to obtain correct estimations, the chain matrices of individual ear canals have to be determined. Best estimates are obtained using the "reflectance phase method," but the "pressure minima method" also provides surprisingly good results, except in narrow frequency ranges. The reflectance phase method is checked using a physical model of the ear canal and the middle ear. The resulting errors of estimation remain within a limit of 3 dB up to more than 10 kHz.

Humans↗