Showing papers by "Free University of Berlin published in 1995"

Computing the fréchet distance between two polygonal curves

Abstract: As a measure for the resemblance of curves in arbitrary dimensions we consider the so-called Frechet-distance, which is compatible with parametrizations of the curves. For polygonal chains P and Q consisting of p and q edges an algorithm of runtime O(pq log(pq)) measuring the Frechet-distance between P and Q is developed. Then some important variants are considered, namely the Frechet-distance for closed curves, the nonmonotone Frechet-distance and a distance function derived from the Frechet-distance measuring whether P resembles some part of the curve Q.

Two‐dimensional electrophoresis of proteins: An updated protocol and implications for a functional analysis of the genome

Abstract: The two-dimensional electrophoresis (2-DE) technique developed by Klose in 1975 (Humangenetik 1975, 26, 211-234), independently of the technique developed by O'Farrell (J. Biol. Chem. 1975, 250, 4007-4021), has been revised in our laboratory and an updated protocol is presented. This protocol is the result of our experience in using this method since its introduction. Many modifications and suggestions found in the literature were also tested and then integrated into our original method if advantageous. Gel and buffer composition, size of gels, use of stacking gels or not, necessity of isoelectric focusing (IEF) gel incubation, freezing of IEF gels or immediate use, carrier ampholytes versus Immobilines, regulation of electric current, conditions for staining and drying the gels - these and other problems were the subject of our concern. Among the technical details and special equipment which constitute our 2-DE method presented here, a few features are of particular significance: (i) sample loading onto the acid side of the IEF gel with the result that both acidic and basic proteins are well resolved in the same gel; (ii) use of large (46 x 30 cm) gels to achieve high resolution, but without the need of unusually large, flat gel equipment; (iii) preparation of ready-made gel solutions which can be stored frozen, a prerequisite, among others, for high reproducibility. Using the 2-DE method described we demonstrate that protein patterns revealing more than 10 000 polypeptide spots can be obtained from mouse tissues. This is by far the highest resolution so far reported in the literature for 2-DE of complex protein mixtures. The 2-DE patterns were of high quality with regard to spot shape and background. The reproducibility of the protein patterns is demonstrated and shown to be thoroughly satisfactory. An example is given to show how effectively 2-DE of high resolution and reproducibility can be used to study the genetic variability of proteins in an interspecific mouse backcross (Mus musculus x Mus spretus) established by the European Backcross Collaborative Group for mapping the mouse genome. We outline our opinion that the structural analysis of the human genome, currently pursued most intensively on a worldwide scale, should be accompanied by a functional analysis of the genome that starts from the proteins of the organism.

Cloning and characterization of a g-protein-activated human phosphoinositide-3 kinase

Abstract: Phosphoinositide-3 kinase activity is implicated in diverse cellular responses triggered by mammalian cell surface receptors and in the regulation of protein sorting in yeast. Receptors with intrinsic and associated tyrosine kinase activity recruit heterodimeric phosphoinositide-3 kinases that consist of p110 catalytic subunits and p85 adaptor molecules containing Src homology 2 (SH2) domains. A phosphoinositide-3 kinase isotype, p110 gamma, was cloned and characterized. The p110 gamma enzyme was activated in vitro by both the alpha and beta gamma subunits of heterotrimeric guanosine triphosphate (GTP)-binding proteins (G proteins) and did not interact with p85. A potential pleckstrin homology domain is located near its amino terminus. The p110 gamma isotype may link signaling through G protein-coupled receptors to the generation of phosphoinositide second messengers phosphorylated in the D-3 position.

Use of immunohistologic and in situ hybridization techniques in the examination of sacroiliac joint biopsy specimens from patients with ankylosing spondylitis

Abstract: Objective To investigate mechanisms involved in inflammation and new bone formation in the sacroiliac (SI) joints of patients with ankylosing spondylitis (AS). Patients and methods Computed tomography-assisted biopsy of the SI joint was performed in 5 patients with AS with a mean disease duration of 4.5 years and radiographic stage 2-3 disease. Immunohistologic studies were performed with the alkaline phosphatase-anti-alkaline phosphatase technique, and cytokine messenger RNA (mRNA) was detected by in situ hybridization. Results Dense cellular infiltrates with varying amounts of CD3+ cells (mean +/- SD 53.3 +/- 24.1%), CD4+ cells (29.7 +/- 17.6%), CD8+ cells (15.8 +/- 11.4%), CD14+ cells (23.6 +/- 16.9%), CD45RO+ cells (48.4 +/- 23.6%), and CD45RA+ cells (4.5 +/- 2.9%) were found in the synovial portion of the SI joints of all 5 patients. In these infiltrates a high amount of tumor necrosis factor alpha (TNF alpha) mRNA and, near the site of new bone formation, a lower amount of transforming growth factor beta (TGF beta) mRNA, were detected, while no message for interleukin-1 was found in the 3 patients examined by this technique. Conclusion The presence of T cells and macrophages was demonstrated in cellular infiltrates in the SI joints of 5 patients with active AS. The finding of abundant TNF alpha message in these joints could have implications regarding potential immunotherapeutic approaches to this disease. TGF beta might be involved in new bone formation in AS.

Learning and memory in the honeybee

Abstract: The understanding of the physiology of learning is dominated by two basically different hypotheses. The deterministic view, following Hebb’s (1949) concept of the memory engram, presupposes a memory groove which is built during memory formation by the adaptive change of a relatively small number of reacting sites or switch points. These so-called ’switchpoint theories’ or ‘place theories’ assume that memory involves a discrete set of cells reserved for the special function of information storage (Young 1964; Eccles 1964; Ungar 1970). The non-deterministic or statistical theory is based on Lashley’s (1950) findings which suggest that all, or nearly all, stored information is distributed throughout the whole association cortex rather than by distinct association paths or centres. The individual neuronal switch points may then be involved in the storage of many different memory traces (John 1967, 1972). The two views are similar in that they take the adaptivity of single synapses between neurones as the basic modifiable component of the nervous system (Eccles and McIntyre 1953; Eccles 1964; Ungar 1970; John 1972). They differ, however, in their conception of the gross structure of the memory system. The crucial problem, then, is to locate the stored information. The spatio-temporal pattern of activity during memory formation produces a localised change in the excitability of specific neurones. It should be possible to find such neurones using the same techniques as have been employed for the location of units in the sensory integration centres.

Treadmill Training With Partial Body Weight Support Compared With Physiotherapy in Nonambulatory Hemiparetic Patients

Abstract: Background and Purpose Treadmill training with partial body weight support is a new and promising therapy in gait rehabilitation of stroke patients. The study intended to investigate its efficiency compared with gait training within regular physiotherapy in nonambulatory patients with chronic hemiparesis. Methods An A-B-A single-case study design compared treadmill training plus partial body weight support (A) with physiotherapy based on the Bobath concept (B) in seven nonambulatory hemiparetic patients. The minimum poststroke interval was 3 months, and each treatment phase lasted 3 weeks. Variables were gait ability assessed by the Functional Ambulation Category, other motor functions tested by the Rivermead Motor Assessment, muscle strength assessed by the Motricity Index, muscle tone rated by the Modified Ashworth Spasticity Scale, and gait cycle parameters. Results Treadmill training was more effective with regard to restoration of gait ability (P<.05) and walking velocity (P<.05). Other motor functio...

Link between the angiographic substudy and mortality outcomes in a large randomized trial of myocardial reperfusion : importance of early and complete infarct artery reperfusion

Abstract: Background The Global Utilization of Streptokinase and TPA for Occluded Coronary Arteries (GUSTO-I) trial was designed to test whether thrombolytic strategies achieving more complete, early, sustained coronary artery patency would lead to further reductions in mortality in patients with acute myocardial infarction. An angiographic substudy within GUSTO-I provided a unique opportunity to examine the relation between mortality and degrees of patency among the regimens. Methods and Results Four thrombolytic strategies were compared in 41 021 patients in GUSTO-I: streptokinase with subcutaneous or intravenous heparin, accelerated tissue plasminogen activator (TPA) with intravenous heparin, and combination streptokinase plus TPA with intravenous heparin. Accelerated TPA was associated with lower 30-day mortality (6.3%) than the other strategies (7.2%, 7.4%, and 7.0%, respectively). Among the 1210 patients in the angiographic substudy randomized to angiography 90 minutes after starting treatment, there was improved patency, particularly Thrombolysis in Myocardial Infarction (TIMI) grade 3 flow, with accelerated TPA over the other regimens ( P <.0001). Coronary artery perfusion (TIMI grade 3) at 90 minutes was also a significant predictor of 30-day survival ( P <.01). To determine whether differences in mortality among the four strategies matched differences in 90-minute patency, a model was developed for predicting mortality differences in the main trial from the angiographic substudy. The model assumed that any differences in treatment effects on 30-day mortality were mediated through differences in 90-minute patency for the four treatments. The predicted rates were then compared with observed mortality rates of the remaining patients in the main trial for each treatment group. The predicted and observed 30-day mortality rates of the four treatments were streptokinase with subcutaneous heparin, 7.46% versus 7.28%; streptokinase with intravenous heparin, 7.26% versus 7.39%; accelerated TPA, 6.31% versus 6.37%; and streptokinase plus TPA, 6.98% versus 6.96%. The correlation between predicted and observed results was .97, and the proportion of squared error explained ( R 2) was .92. Conclusions The close relation between the predicted and observed 30-day mortality rates supports the concept that an important mechanism for improved survival with thrombolytic therapy is achievement of early, complete perfusion. The close match provides a strong biological explanation for the mortality differences seen in GUSTO-I and a sound rationale for the additional survival advantage of the accelerated TPA regimen. Irrespective of which treatment is used, early and complete restoration of infarct artery perfusion represents an essential goal of myocardial reperfusion therapy.

Lipids in biological membrane fusion.

Abstract: The results reviewed suggest that membrane fusion in diverse biological fusion reactions involves formation of some specific intermediates: stalks and pores. Energy of these intermediates and, consequently, the rate and extent of fusion depend on the propensity of the corresponding monolayers of membranes to bend in the required directions. Proteins and peptides can control the bending energy of membrane monolayers in a number of ways. Monolayer lipid composition may be altered by different phospholipases [50, 85, 90], flipases and translocases [4, 50]. Proteins and peptides can change monolayer spontaneous curvature or hydrophobic void energy by direct interaction with membrane lipids [20, 32, 111]. Proteins may also provide some barriers for lipid diffusion in the plane of the monolayer [83, 141]. If diffusion of lipids at some specific membrane sites (e.g., in the vicinity of fusion protein) is somehow hindered, the energy of the bent fusion intermediates would reflect the elastic properties of these particular sites rather than the spontaneous curvature of the whole monolayers. Proteins may deform membranes while bringing them locally into close contact. The alteration of the geometric (external) curvature will certainly change the elastic energy of the initial state and, thus affect the energetic barriers of the formation of the intermediates [143]. In addition, the area and the energy of the stalk can be reduced by preliminary bending of the contacting membranes [111]. The possible effects of proteins and polymers on local elastic properties and local shapes of the membranes have been recently analyzed [22, 39, 45, 63]. These studies may provide a good basis for future development of theoretical models of protein-mediated fusion. Various models for biological fusion have been presented as hypothetical sequences of intermediate conformations of proteins, with membrane lipids just covering the empty spaces between the proteins. Although the results discussed above do not allow us to draw an allexplaining cartoon of the fusion mechanism, they do indicate which properties of membrane lipid bilayers (if modified by fusion proteins) would get these bilayers to fuse. In addition, these data suggest a specific geometry to bent fusion intermediates (stalks and pores) and imply a contribution by lipids to the energy of these intermediates. We think that the synthesis of rapidly developing structural information on fusion proteins with the analysis of the physics of membrane rearrangement may soon yield a real understanding of the fascinating and fundamental phenomenon of membrane fusion.

Long Memory in Inflation Rates: International Evidence

Abstract: We examine monthly inflation rates of five industrial countries. The application of tests against stationarity as well as tests against a unit root yield contradictory results. Thus fractional integration allowing for long memory is a plausible model. We discuss and apply the periodogram regression to estimate the difference parameters. For all countries we find estimates significantly different from 1 as well as from 0. This is evidence in favor of long memory. Specification tests and maximum likelihood estimates support the fitted models. Finally, we relate our empirical results to the construction of the data.

Malignant liver tumors treated with MR imaging-guided laser-induced thermotherapy : technique and prospective results

Abstract: PURPOSE: To evaluate magnetic resonance (MR) imaging-guided laser-induced thermotherapy (LITT) of liver metastases. MATERIALS AND METHODS: In a phase II study, 20 patients with 33 metastases from colorectal carcinoma (75%) or other primary tumors (25%) underwent LITT. MR thermometry performed with fast low-angle shot sequences was used to monitor therapy on-line, and dynamic and static contrast material-enhanced MR images enabled estimation of the degree of resultant necrosis. Follow-up studies were performed 3 months after thermotherapy. RESULTS: The thermosequences enabled accurate on-line monitoring in 85% of lesions. In 69% of lesions 20 mm in diameter or smaller, contrast-enhanced MR images depicted substantial necrosis, with a local tumor control rate of 69% after 6 months and 44% after 12 months. Among lesions larger than 20 mm, necrosis was frequently incomplete, with a local control rate of only 41% after 6 months and 27% after 12 months. CONCLUSION: MR imaging-guided LITT of liver metastases is ...

Colour preferences of flower-naive honeybees

Abstract: Flower-naive honeybees Apis mellifera L. flying in an enclosure were tested for their colour preferences. Bees were rewarded once on an achromatic (grey, aluminium or hardboard), or on a chromatic (ultraviolet) disk. Since naive bees never alighted on colour stimuli alone, a scent was given in combination with colour. Their landings on twelve colour stimuli were recorded. Results after one reward (“first test”) were analysed separately from those obtained after few rewards (“late tests”). 1) After pre-training to achromatic signals, bees preferred, in the first test, bee-uv-blue and bee-green colours. With increasing experience, the original preference pattern persisted but the choice of bee-blue and bee-green colours increased. 2) Neither colour distance of the test stimuli to the background or to the pre-training signal, nor their intensity, nor their green contrast, accounted for the colour choice of bees. Choices reflected innate preferences and were only associated with stimulus hue. 3) Bees learned very quickly the pre-trained chromatic stimulus, the original colour preferences being thus erased. 4) Colour preferences were strongly correlated with flower colour and its associated nectar reward, as measured in 154 flower species. 5) Colour preferences also resemble the wavelength dependence of colour learning demonstrated in experienced bees.

Design principles of vascular beds.

Abstract: Hemodynamic parameters were determined in each vessel segment of six complete microvascular networks in the rat mesentery by using a combination of experimental measurements and theoretical stimulations. For a total number of 2592 segments, a strong unified dependence of wall shear stress on intravascular pressure for arterioles, capillaries, and venules was obtained. All three types of segments exhibit an essentially identical variation of shear stress from high to low values (from approximately 100 to 10 dyne/cm2) as intravascular pressure falls from 70 to 15 mm Hg. On the basis of these observations, it is proposed that vascular beds grow and adapt so as to maintain the shear stress in each vessel at a level that depends on local transmural pressure. In contrast to Murray's classic 'minimum-cost' hypothesis, which implies uniformity of wall shear rate throughout the vasculature, the proposed design principle provides an explanation for the functionally important arteriovenous asymmetry of wall shear rates and flow resistance in the circulation.

Hematopoietic rescue after high-dose chemotherapy using autologous peripheral-blood progenitor cells or bone marrow: a randomized comparison.

Abstract: PURPOSETo compare autologous bone marrow (BM) with peripheral-blood progenitor cells (PBPC) as hematopoietic rescue after high-dose chemotherapy (HDCT).PATIENTS AND METHODSFrom January 1991 until April 1993, 47 consecutive patients with relapsed or refractory germ cell tumors were randomized to either BM harvest or collection of PBPC mobilized by chemotherapy plus granulocyte colony-stimulating factor (G-CSF). After additional conventional-dose salvage treatment, all patients received HDCT with carboplatin 1,500 mg/m2, etoposide 2,400 mg/m2, and ifosfamide 10 g/m2 with either BM or PBPC rescue.RESULTSForty-six patients were assessable for hematologic reconstitution, and one patient died on day +4 before engraftment. Rescue using PBPC resulted in a significantly shorter recovery time to neutrophil counts more than 500/microL (10.0 v 11.0 days, P < .01), neutrophil counts more than 1,000/microL (10.0 v 12.0 days, P = .001), and platelet counts more than 20,000/microL (10.0 v 17.0 days, P < .01), as well as ...

The cobalt, zinc, and cadmium efflux system CzcABC from Alcaligenes eutrophus functions as a cation-proton antiporter in Escherichia coli.

Abstract: The function of the CzcABC protein complex, which mediates resistance to Co2+, Zn2+, and Cd2+ in Alcaligenes eutrophus by cation efflux, was investigated by using everted membrane vesicles of Escherichia coli and an acridine orange fluorescence quenching assay. Since metal cation uptake could not be measured with inside-out membrane vesicles prepared from A. eutrophus and since available E. coli strains did not express the Czc-mediated resistance to cobalt, zinc, and cadmium salts, mutants of E. coli which exhibited a Czc-dependent increase in heavy metal resistance were isolated. E. coli mutant strain EC351 constitutively accumulated Co2+, Zn2+, and Cd2+. In the presence of Czc, net uptake of these heavy metal cations was reduced to the wild-type level. Inside-out vesicles prepared from E. coli EC351 cells displayed a Czc-dependent uptake of Co2+, Zn2+, and Cd2+ and a cation-triggered acridine orange fluorescence increase. The czc-encoded protein complex CzcABC was shown to be a zinc-proton antiporter.

Presence of Wilms' Tumor Gene (wt1) Transcripts and the WT1 Nuclear Protein in the Majority of Human Acute Leukemias

Abstract: The wt1 gene is located on chromosome 11p13 and encodes a zinc finger motif-containing transcription factor involved in regulation of growth and differentiation. Its expression was shown during embryonic development in various tissues as well as in a few human malignancies including acute leukemias. Using RT-PCR, we found wt1 gene expression in blast cells of the majority of 150 acute leukemia patients. Particularly, the wt1 transcript was detected in 12 of 14 (86%) pre-pre-B-ALL patients, in 33 of 41 (80%) cALL patients, in 23 of 31 (74%) T-ALL patients, and in 53 of 57 (93%) AML patients. Additionally, mononuclear cells from CML patients expressed the wt1 gene only when diagnosed with blast crisis. In contrast to acute human leukemias, mononuclear cells from reactive bone marrow (n = 4), and peripheral blood of healthy volunteers (n = 20), as well as normal peripheral CD34+ hematopoietic progenitors (n = 6) did not express the wt1 gene at detectable levels. Using the anti-WT1 MoAb 6F-H2 in an immunofluorescence assay on single cell level, we found the translated WT1 protein only in nuclei of leukemia blast cells but not in nuclei of normal CD34+ hematopoietic progenitor cells. Blast cells of 12 of 20 leukemia patients (60%) all tested positive for the wt1 gene expression by RT-PCR displayed a strong nuclear immunofluorescence. Its expression in the majority of human acute leukemias but not in normal mononuclear blood cells and normal CD34+ hematopoietic progenitors qualifies the wt1 gene transcript as a 'pan-acute leukemic' marker probably useful in monitoring minimal residual disease after chemotherapy and in detecting leukemic blast cells in purged or unpurged hematopoietic stem cell preparations intended to be used for autologous bone marrow transplantation.

Quantum Control of Wave Packet Evolution with Tailored Femtosecond Pulses.

Abstract: Using theory to guide the choice of pulse shape, we have synthesized frequency-chirped laser pulses and used them to control the evolution of vibrational wave packets on the $B$ excited state of iodine. A negatively chirped pulse produces a wave packet at the target time localized about an internuclear position and momentum of our choice. An approximately time-reversed pulse, however, produces a delocalized wave packet. The experimental results are in very good qualitative agreement with quantum simulations.

A soluble form of TRAP (CD40 ligand) is rapidly released after T cell activation.

Abstract: TRAP is a tumor necrosis factor (TNF)-related, 33-kDa type II transmembrane protein almost exclusively expressed on the surface of activated CD4+ T lymphocytes. Interaction of TRAP with CD40 on B cells is of paramount importance for immunoglobulin class switching and subsequent synthesis of IgG, IgA or IgE in vivo. We now provide evidence that activated T cells not only express cell membrane-associated TRAP but also a soluble form of TRAP (sTRAP). After generating monoclonal antibodies against TRAP and establishing a TRAP-specific enzyme-linked immunosorbent assay we were able to detect substantial amounts of sTRAP in the supernatants of activated T cells. The onset and rate of sTRAP release was found to parallel the expression of TRAP on the cell surface. sTRAP, an 18-kDa protein, is generated by proteolytic processing of full-length TRAP in an intracellular compartment. Starting with methionine 113 of full-length TRAP, sTRAP lacks the transmembrane region and a part of the extracellular domain but contains the entire TNF-alpha homology region and can, therefore, bind to CD40. Like other members of the TNF superfamily (e.g. TNF-alpha, Fas/APO-1 ligand), TRAP thus has the potential to be biologically active not only in a transmembrane form but also as a soluble molecule.

Expression of the bcl-2 gene family in normal and malignant breast tissue: low bax-alpha expression in tumor cells correlates with resistance towards apoptosis.

Abstract: We have studied the expression of the apoptosis-regulating genes bcl-2, bcl-x, bax and APO-1/fas (CD95) in human breast cancer. The expression pattern of these genes in human breast-cancer tissues and breast-cancer-derived cell lines was compared to that seen in normal breast epithelium and breast epithelial cell lines. No difference with regard to bcl-2 and bcl-xL expression was observed between normal breast epithelium and tumor tissue or breast cancer and non-malignant epithelial cell lines. In contrast, bax-alpha, a splice variant of bax, which promotes apoptosis, is expressed in high amounts in normal cell lines and breast tissue, whereas only weak or no expression could be detected in cancer-cell lines and malignant tissue. In contrast to malignant cell lines, which express low levels of bax-alpha, non-malignant epithelial cell lines displaying high amounts of bax-alpha were highly sensitive to induction of programmed cell death by both serum starvation and APO-1/fas (CD95) triggering. We therefore propose that dysregulation of apoptosis contributes to the pathogenesis of breast cancer, at least in part, due to an imbalance between anti-apoptosis genes (such as bcl-2/bcl-x) and apoptosis-promoting genes (bax).

Effects of Treatment and Storage Conditions on Ceramic/Composite Bond Strength

Abstract: During the past few years, the interest in using ceramic inlays and veneers has increased. New materials and methods have been introduced to bond these restorations to resinous materials. Since our knowledge of how to optimize such bonding is limited, the objective of this study was to test the hypothesis that various surface treatment variables and combinations of these variables affect the strength of the ceramic/composite interphase of ceramic inlays differently. The influences of material composition, surface-roughening method, silane treatment, silane heat treatment, and storage condition on bond strength were investigated. Three ceramics (Dicor, Mirage, Vitabloc), three surface-roughening methods (etching, sandblasting, grinding), three silane treatments (gamma-methacryloxypropyltrimethoxysilane [MPS], MPS+paratoluidine, vinyltrichlorosilane), two heat treatments (20 degrees C for 60 s, 100 degrees C for 60 s), and two storage conditions (24-hour dry, one yr in water at 37 degrees C) were studied. For each of the 108 combinations, five specimens were tested. Ceramic cylinders were treated according to group assignment and bonded to blocks of the same ceramic material with a dual-cured resin. The shear bond strength was determined, and the experimental factors were evaluated by analysis of variance. The results showed that surface-roughening method had the strongest effect on bond strength, while ceramic selection had the least significant effect. Of the surface-roughening methods, etching was associated with higher bond strength values than either sandblasting or grinding.(ABSTRACT TRUNCATED AT 250 WORDS)