A competitive immunoassay for cortisol based on capillary electrophoresis (CE) and laser-induced fluorescence is described. The work involved the production of assay reagents and the development of separation conditions allowing for routine analysis of serum samples. Fluorescein-labeled cortisol was synthesized and purified. Fab fragments were produced from mouse monoclonal anticortisol antibody and purified using a POROS cation exchange chromatography column. After incubation of these reagents with serum, free and bound labeled antigen were separated by CE with high reproducibility. No prior sample cleanup of the serum samples was necessary. Serum calibration curves were established and used for the quantitation of cortisol in serum. The results demonstrate feasibility for a cortisol assay based on CE operating directly on serum samples.

Electrophoresis↗

PubMed1995

Origin of P16 median nerve SEP component identified by dipole source analysis--subthalamic or within the thalamo-cortical radiation?

Following median nerve stimulation, several monophasic peaks were recorded at the scalp in the 15-18 ms time range. Source analysis, using three different methods, modelled a source near the centre of the head with an orientation towards the activated hemisphere and a peak activity at 16 ms post stimulus. Magnetic recordings detected no signal in this time range, which confirmed a subcortical location of the source. From dipole localization it was not possible to assign the exact origin of the P16 source to either the subthalamic level or the thalamo-cortical radiation, because of the limited spatial resolution at the centre of the spherical head model. An estimate of the conduction velocity of the medial lemniscus pointed towards a subthalamic origin. The P16 source was preserved in two patients with a lesion of the thalamo-cortical radiation and the ventral thalamus. Further evidence for a subthalamic location of P16 was derived from the physical mechanisms generating far-field potentials.

Adult↗

PubMed1995

Comparison of realistically shaped boundary-element and spherical head models in source localization of early somatosensory evoked potentials.

Source localizations of early somatosensory evoked potentials and electrical potentials produced by dipoles in the region of the central sulcus were computed using realistically shaped boundary-element head models (BEM) and compared to localizations obtained using 3-shell spherical models. Realistically shaped 3-shell boundary-element-models were constructed on the basis of the individual anatomy obtained from 3D-MR-tomography in 6 subjects. Spherical head models were fitted to the actual locations of the electrodes and to the surface of the heads, respectively. Source locations calculated within the spherical head models differed by an average of 4 mm (range: 2 to 7 mm) with respect to the 3-shell BEM, taking into account the limited accuracy of this model. This mislocation was most prominently due to deeper source locations predicted using a spherical head model and caused by incorrect modelling of the geometry of the heads, although sources were located in a favourable region of the heads.

Adult↗

PubMed1995

Coordinate system matching for neuromagnetic and morphological reconstruction overlay.

The overlay of functional and morphological images is an essential tool for advanced and improved functional diagnosis showing the correlation between spatial structures or lesions and functional areas. We present an improved coordinate system matching technique. The well known method of three orthogonal coils combined in one coilset for an angular-independent measure is validated for the use of first- or higher order gradiometers instead of magnetometers. The coilset localization procedure was modified with lock-in detection and current feedback for better long range sensitivity. Real measurements with the 31-channel Philips-MEG system have been carried out. A very good localization accuracy below the measuring area with deviations below 2 mm was found. For coordinate system matching, a 3-D cursor with surface images from segmented MR-data was implemented and an optimized, weighted least squares fit transformation algorithm between functional and morphological systems was developed. The resulting transformations consist of weighted shifts and best-fit rotations and lead to deviations of marker positions in the mm range, depending mainly on the spatial accuracy of the marker fixation.

Algorithms↗

PubMed1995

Women in dermatology. XI.

Explore the source record for details and available documents.

Dermatology↗

PubMed1995

The role of macrophage receptors in adhesion and uptake of Leishmania mexicana amastigotes.

Amastigotes of the protozoan parasite Leishmania proliferate in phagolysosomes of mammalian macrophages. Propagation of the infection is considered to occur by host-cell rupture and uptake of released parasites by uninfected macrophages. In this study, the kinetics of binding of L mexicana mexicana amastigotes to COS cells and to COS cells transfected with three different macrophage receptors (FcRII-B2, receptor for the Fc-domain of immunoglobulins; CR3, complement type 3 receptor and the mannose receptor) is compared to the rate of adhesion to peritoneal macrophages. Amastigotes isolated from macrophages cultivated in vitro bind with slow, sigmoid kinetics to COS cells expressing either of the three receptors, or to peritoneal macrophages. In contrast, amastigotes isolated from mouse lesions bind with rapid, hyperbolic kinetics to COS cells expressing the Fc receptor or to peritoneal macrophages but with slow, sigmoid kinetics to COS cells expressing the CR3 or the mannose receptor. As shown by immunofluorescence experiments, lesion-derived amastigotes contain host-derived immunoglobulins (Ig) but no complement component 3 at their surface. It is concluded that amastigotes contain no intrinsic ligand at their surface, which enables high-affinity interactions with macrophages. Opsonization by specific Ig may be of relevance in vivo because firstly, in cryosections of mouse lesions extracellular amastigotes containing surface Ig can be detected and, secondly, B cell-deficient mice reconstituted with parasite-specific Ig show a modest increase in the rate of lesion development. In addition, it is shown that amastigotes are internalized by COS cells and grow in large parasitophorous vacuoles similar to those observed in macrophages.

Adhesiveness↗

PubMed1995

[Effect of various methods of resection of the stomach in liberation of cholecystokinin, neurotensin and on pancreatic function].

In a follow up study 19 patients after Billroth-I, 22 patients after Billroth-II-resection and 38 patients after total gastrectomy (23 with Roux-Y-reconstruction and 15 with Longmire-Gütgemann) underwent a stool fat determination and an indirect pancreatic function test with fluorescein dilaurate in serum und urine. In 9 of 19 cases (47.3%) after B-I-resection and 14 of 22 patients (63.5%) after B-II-resection there were pathological results of the PLT-test in urine. After total gastrectomy as well with reconstruction of the duodenal passage as with its exclusion the PLT-test results were pathological in 60% resp. 87%. These results indicate a secondary pancreatic insufficiency following gastric resection.

Adult↗

PubMed1995

Solution-phase immunoassay for determination of cortisol in serum by capillary electrophoresis.

We describe a competitive solution-phase immunoassay for serum cortisol determination that involves capillary electrophoresis (CE) combined with laser-induced fluorescence for separation and quantification. A polyclonal antibody preparation and fluorescein-labeled cortisol are used as assay reagents. 8-Anilino-1-naphthalenesulfonic acid was added to the serum sample during the assay to promote the release of cortisol from endogenous binding proteins. Conditions for the rapid separation of free and bound labeled antigen by CE were developed. Aspects of assay performance are evaluated in this report. The resulting assay protocol allows for the analysis of serum samples without extraction or other sample preparation steps.

Anilino Naphthalenesulfonates↗

PubMed1995

Deoxycholate and cholate modulate the source of cholesterol substrate for bile acid synthesis in the rat.

In the current study, the role of the supply of preformed and newly synthesized cholesterol for the feedback control of the synthesis of different bile acids and the secretion of biliary cholesterol was investigated. To define these cholesterol fluxes and the possibility of a different modulation by bile acids with different suppressive capacities, a continuous labeling with tritiated water was used in rats with an extracorporeal bile duct receiving intraduodenal infusions of taurocholate or taurocholate plus deoxycholate. After bile acid pool depletion (6 to 9 hours) total muricholate, cholate, and chenodeoxycholate synthesis was variably increased (24% to 93%) during an infusion of 304 mumol taurocholate/kg per hour. The increase in bile acid synthesis and biliary cholesterol output was predominantly due to the utilization of preformed (unlabeled) cholesterol. The addition of 52 mumol/kg per hour of deoxycholate to 258 mumol/kg per hour of taurocholate had a comparable effect. In the late period (30 to 54 hours), the taurocholate infusion had little impact on total muricholate and chenodeoxycholate synthesis but caused by a significant increase of the proportion from performed cholesterol. Both total cholate production and its synthesis from de novo (labeled) cholesterol was inhibited by 30% (P < .05) and 64% (P < .01), respectively. The secretion rate of total and de novo biliary cholesterol was higher (65% and 72%; P < .01) compared with controls. In comparison, the combined bile acid infusion led to a further increase of total muricholate synthesis (P < .05), which was again due to an enhanced synthesis from performed cholesterol (P < .001). Similar changes were observed in chenodeoxycholate. The more pronounced suppression of total cholate synthesis by 81% (P < .05) was due to a diminished cholate synthesis from both de novo cholesterol by 72% (P < .001) and preformed cholesterol by 91% (P > .05). We conclude that the modulation of the synthesis of the various primary bile acids in the rat differs and feedback regulation of cholate synthesis by taurocholate and deoxycholate is mediated by different mechanisms of control, including inhibition of cholesterol 7 alpha-hydroxylase, HMG-CoA reductase, and uptake of lipoprotein cholesterol.

Animals↗

PubMed1994

Charged particle spectra in central S+S collisions at 200 GeV/c per nucleon.

Explore the source record for details and available documents.

Journal Article↗

PubMed1994

[Not Available].

Historical sociobiographic accounts on members of the scientific and technical professions in the years of the Weimar Republic and after, are as yet scarce. This applied notably to women in the scientific community. Having formally been admitted to academic studies at German Universities only in 1908, their claims of wanting to apply their newly gained knowledge and to pursue academic careers were still not unquestioned by society. The social and cognitive integration of "the female" in male dominated science organisation, especially in the natural sciences and their kin fields in industry, remains problematic to-day. Isolde Hausser, daughter of the ambitious but little succesful inventor-entrepreneur Hermann Ganswindt, took her doctoral degree in physics at Berlin University in 1914, then worked as head of a group at a "Telefunken" laboratory for vacuum tubes till 1929, before she became research scientist at the Kaiser Wilhelm Institute for Medical Research, Heidelberg. There she worked on photoerythemaes and the formation of pigment and discovered the specific action of longwave ultraviolet. She contributed important results to our knowledge on the constitution and the behaviour of organic compounds by modern physical methods. She died of cancer on 5th October 1951.

Academies and Institutes↗

PubMed1994

Disruption of the gene encoding the EcmA, extracellular matrix protein of Dictyostelium alters slug morphology.

The ecmA and ecmB genes of Dictyostelium are expressed in prestalk and stalk cells. They encode components of the slime sheath, the extracellular matrix that surrounds the migrating slug, and the stalk tube, the matrix that encases stalk cells. We have generated, by homologous gene disruption, a mutant in which the ecmB gene is inactivated but the strain develops normally. In contrast, ecmA null mutant strains develop to form abnormally long and thin standing slugs. While the slime sheath of mutant slugs appears to be normal in electron microscopic observations, the sheath material remaining on the substratum after the slug travels through it is abnormally susceptible to breakage. After a short period of migration the axial ratio of mutant slugs decreases to that of normal slugs and, at culmination, normal fruiting bodies are produced. These data suggest that the EcmA protein has its primary role during slug formation, where it contributes to the strength of the slime sheath, and that the function of the EcmB protein is dispensible.

Animals↗

PubMed1994

Source analysis of median nerve and finger stimulated somatosensory evoked potentials: multichannel simultaneous recording of electric and magnetic fields combined with 3D-MR tomography.

At the current state of technology, multichannel simultaneous recording of combined electric potentials and magnetic fields should constitute the most powerful tool for separation and localization of focal brain activity. We performed an explorative study of multichannel simultaneous electric SEPs and magnetically recorded SEFs. MEG only sees tangentially oriented sources, while EEG signals include the entire activity of the brain. These characteristics were found to be very useful in separating multiple sources with overlap of activity in time. The electrically recorded SEPs were adequately modelled by three equivalent dipoles located: (1) in the region of the brainstem, modelling the P14 peak at the scalp, (2) a tangentially oriented dipole, modelling the N20-P20 and N30-P30 peaks, and part of the P45, and (3) a radially oriented dipole, modelling the P22 peak and part of the P45, both located in the region of the somatosensory cortex. Magnetically recorded SEFs were adequately modelled by a single equivalent dipole, modelling the N20-P20 and N30-P30 peaks, located close to the posterior bank of the central sulcus, in area 3b (mean deviation: 3 mm). The tangential sources in the electrical data were located 6 mm on average from the area 3b. MEG and EEG was able to locate the sources of finger stimulated SEFs in accordance with the somatotopic arrangement along the central fissure. A combined analysis demonstrated that MEG can provide constraints to the orientation and location of sources and helps to stabilize the inverse solution in a multiple-source model of the EEG.

Adult↗

PubMed1994

Distribution of parasite cysteine proteinases in lesions of mice infected with Leishmania mexicana amastigotes.

It is well established that Leishmania mexicana amastigotes contain large amounts of cysteine proteinases in their extended lysosomes. In this study it is shown that the cell-free supernatant of homogenized lesion tissue from infected mice contains large amounts of acid proteinases. The majority of this enzymatic activity also corresponds to cysteine proteinases from L. mexicana amastigotes. Immunoelectron microscopy of mouse lesion sections suggests, that frequently amastigotes lyse and release lysosomal cysteine proteinases into the parasitophorous vacuole of infected macrophages. The cysteine proteinases are also found extracellularly in the tissue presumably as a result of macrophage rupture and appear to persist in the lesion tissue, where they may damage host cells and the extracellular matrix.

Amino Acid Sequence↗