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Biomedical subjects

R Langer

Publications and source records attributed to R Langer.

At least 181 records · Page 10Linked to original sources
PubMed1997

[Value of lung filter, asymmetric film-screen technique and digital storage screen radiography in diagnosis of coin lesion].

AIM: To evaluate the image quality of an asymmetric film-screen combination (a-FSC), a conventional screen-film combination (FSC) at speed class 200, a lung filter, and digital luminescence radiograms in detecting pulmonary nodules. METHODS: Detail perception studies were carried out with an anthropomorphic phantom. The image systems were exposed under standardised conditions with 125 kVp. The systems' perception efficiency was evaluated on the basis of 12.240 single observations with ROC analysis. RESULTS: In the overall evaluation the a-FSC and the filter achieved equally high assessments of 0.878 +/- 0.018 and 0.860 +/- 0.21 respectively (p > 0.05). The 200 speed FSC and the storage phosphor screen radiograms did not differ significantly. In the mediastinum all the tested systems were clearly superior to the 200 speed FSC (p < 0.05). In the lung fields on the other hand the 200 speed system was given a higher assessment of 0.866 +/- 0.026. The a-FSC had slight advantages over the lung filter and the digital imagining modes (p > 0.05). CONCLUSIONS: The a-FSC and digital luminescence radiograms provide significantly more diagnostic information in the mediastinum, together with high detail detectability in the lung fields and without requiring additional dosage.

Humans↗
PubMed1997

[Imaging methods in diagnosis of pheochromocytoma].

In the radiological evaluation of adrenal masses today computed tomography (CT) has the highest sensitivity of all imaging modalities for tumor detection. In addition, CT is able to assess potential concomitant processes in the surrounding tissues. Magnetic resonance imaging (MRI) is the method of choice, when adrenal masses cannot be differentiated by CT. When chemical-shift imaging, fat-suppression, and dynamic contrast-enhanced sequences are used for tissue characterization, pheochromocytomas can be distinguished from adenomas. Due to the multiplanar imaging ability MRI is optimal for the preoperative work-up of the anatomical situation in patients with adrenal masses. Extraadrenal paragangliomas can be assessed with MRI, too. Ultrasound is of minor relevance in the assessment of pheochromocytomas and other adrenal masses, as is angiography, which is only carried out in special cases to evaluate the vascular supply.

Adrenal Gland Neoplasms↗
PubMed1997

[Minimally invasive treatment of abscesses by CT-controlled drainage with a basket catheter system].

PURPOSE: To examine whether the percutaneous drainage of abscess formations by a new basket catheter system is usefull. MATERIAL AND METHODS: 58 patients with abscesses in different locations and origins have been treated by an interventional radiologic procedure. On the whole 77 basket-catheters were placed under CT-guidance into abscess formations of different size and localization. 36 patients developed an abscess after surgery, two patients achieved abscess drainage after embolisation of a tumor. In 20 patients the abscess was a complication of a septic infectious disease. RESULTS: Open surgery was avoided in 41/58 patients of these patients 9/41 received only percutaneous drainage and 32/41 patients suffering from illness were given antibiotic medication according to the resistogram in combination with percutaneous drainage. 17/58 patients required secondary surgery but with a lower risk because of the smaller abscess volume and the better clinical constitution after percutaneous drainage. CONCLUSIONS: A minimally invasive management of abscesses using a basket-catheter system is successful even in localisations deep inside the body and hard to reach. An irreversible catheter occlusion followed by the implantation of a new catheter system could be avoided in all patients.

Abdominal Abscess↗
PubMed1997

[Changes in blood rheology and microcirculation after preparatory combined thrombocytapheresis and plasmapheresis].

INTRODUCTION: Because of anticoagulation, changes in blood composition, and--perhaps--extracorporeal circulation, donor apheresis should cause alterations in hemorheology and hence in the perfusion of the microvasculature. MATERIAL AND METHODS: 19 regular blood donors were included. According to our standard protocol for automated collection of blood components with the MCS 3p cell separator, we harvested 1 unit of platelets and 2 units of plasma in each case. Prior to, 1 h after, and 24 h after donation, the following parameters were measured: total serum protein (tsp), hematocrit (hc), whole blood and plasma viscosity (wbv/pv), red cell aggregability (rca) and blood flow velocity of the nail-fold capillaries (bfv). RESULTS: The following parameters decreased 1 h/24 h after donation: tsp (p < 0.001/p = 0.008), elastic wbv (p = 0.018/p < 0.001), viscous wbv (p = 0.85/p = 0.0031), pv (p < 0.001/p < 0.001), static rca (p < 0.001/p = 0.0073), dynamic rca (p < 0.001/p = 0.017). The hc showed an initial increase (p < 0.001) with a subsequent overshooting decrease after 24 h (p < 0.001). 1 h after donation bfv raised (p = 0.0065). It decreased after 24 h and remained only slightly higher than the initial level (p = 0.27). CONCLUSIONS: Automated combined collection of platelets and plasma gives rise to: i) Improvement of rheological properties of the donor's blood and increased bfv of his nail-fold capillaries within the 1st h after apheresis. ii) 24 h after donation the improved hemorheological properties remain demonstrable, but the bfv of nail-fold capillaries declines and shows a trend toward the starting-point. iii) Taken together, this is possibly reflecting adapted hemodynamic and vasoconstrictor regulation for altered hemorheological conditions.

Adaptation, Physiological↗
PubMed1997

[Therapeutic thrombocytapheresis--effect on hemorheologic parameters].

Common features of all myeloproliferative diseases (CMPE) are a markedly increased number of platelets and restrictions in the subjective feeling of the patients, comprising symptoms like nausea, headache, sensory deficits and transient paresis. Meanwhile, it has been shown that platelet pheresis (mTD) does not only reduce platelet counts sufficiently but also results in an impressive relief of the subjective complaints. To test the part that rheological mechanisms play hereby, relevant rheological parameters in the blood of 22 CMPE patients and of 8 healthy platelet donors as controls were analyzed before and after treatment with an AS-104 cell separator. Concerning the viscosity of whole blood und the filterability of erythrocytes, there was seen neither a difference between the patients and the controls, nor before and after mTD. As a result of treatment of patients, the moderately raised plasma viscosity was normalized, whereas the aggregation of erythrocytes was significantly lowered. It is concluded that there is an influence of hemorheological conditions on the patient's subjective feeling. These would originate in neurological dysfunctions caused by CMPE-induced restrictions in cortical microcirculation. The erythrocyte aggregation, lowered by mTD, would cause an improvement of microcirculatory fluidity and would, in consequence, abolish the neurological symptoms.

Blood Viscosity↗
PubMed1997

[Electron microscopy study of HES cryopreserved erythrocytes].

Using electron microscopy (EM), it was examined whether cryopreservation with HES causes shape changes of erythrocytes. Each of 11 erythrocyte suspensions (Hct = 40; HES 200,00/0.5/12.5%; 60 mM NaCl) was separated into 40-ml samples, cooled down to -196 degrees C and finally stored. In addition, 11 samples were stored at -80 degrees C for 3 months. The preparation for EM was done immediately after thawing or in case of native cells shortly after donation. On EM micrographs, there was no visible difference between native and cryopreserved erythrocytes. In every case the preparations showed normocytes, either as single cells or having attracted other ones, forming rouleau. Packed cells were attached tightly to each other without any gap in between. The tangent count method neither revealed an excess of convexity nor of concavity. The erythrocyte membrane looked normal, and the cytoplasmatic space was filled with electron-dense material (hemoglobin) homogeneously; Heinz bodies were not seen. Scanning microscopy portrayed native as well as cryopreserved cells as discocytes with the characteristic bioconcave resting shape of human erythrocytes. It is concluded that cryopreservation of erythrocytes with HES does not cause shape changes. Therefore, a sequestration into the reticulo-endothelial system (RES) by means of identification of morphological abnormalities may not be expected.

Blood Preservation↗
PubMed1997

[CT angiography of the pulmonary artery in patients with bronchial carcinoma].

The aim of the study was to evaluate 3D computed tomography (3D-CT) for the visualization of tumor extent with respect to the infiltration of the pulmonary arteries in patients with locally advanced small and nonsmall-cell lung cancer. A total 61 examinations in 40 patients with bronchial carcinoma were performed with contrast-enhanced (150 ml injected with 4 ml/s after bolus tracking) 2 mm spiral CT (pitch 1.5, increment 1 mm) (Somatom Plus4, Siemens AG, Germany). Using the implemented software cine mode, surface shaded display (SSD), and maximum intensity projection (MIP), reconstructions of the pulmonary arteries and the tumors were generated. In comparison to conventional spiral thoracic CT, the optimized cross-sectional images of the mediastium represented a benefit with the technique described, and this can help to diagnose vessel infiltration and may therefore be of value for pre- and post-therapeutic staging in modern stage-adapted multimodality treatment programs.

Angiography↗
PubMed1996

Tissue-engineered heart valves. Autologous valve leaflet replacement study in a lamb model.

BACKGROUND: We have previously reported the successful creation of tissue-engineered valve leaflets and the implantation of these autologous tissue leaflets in the pulmonary valve position. This study was designed to trace cultured cells that were seeded onto a biodegradable polymer with the use of a 1,1'-dioctadecyl-3,3,3' 3'-tetramethylindo-carbocyanine perchlorate (Di-1) cell-labeling method. We also examined the time-related biochemical, biomechanical, and histological characteristics and evolution of these tissue constructs. METHODS AND RESULTS: Mixed cell populations of endothelial cells and fibroblasts were isolated from explanted ovine arteries. Endothelial cells were selectively labeled with an acetylated low density lipoprotein marker and separated from fibroblasts with the use of a fluorescence-activated cell sorter. A synthetic biodegradable scaffold consisting of polyglycolic acid fibers was seeded first with fibroblasts, then coated with endothelial cells. Using these methods, we implanted autologous cell/polymer constructs in six animals. In two additional control animals, a leaflet of polymer was implanted without prior cell seeding. In each animal, cardiopulmonary bypass was used to completely resect the right posterior leaflet of the pulmonary valve and replace it with an engineered valve leaflet with (n = 6) or without (n = 2) prior cultured cell seeding. The animals were killed either after 6 hours or after 1, 6, 7, 9, or 11 weeks, and the implanted valve leaflets were examined histologically, biochemically, and biomechanically. 4-Hydroxyproline assays were performed to determine collagen content. Leaflet strength was evaluated in vitro with a mechanical tester Factor VIII and elastin stains were done to verify histologically that endothelial cells and elastin, respectively, were present. Animals receiving leaflets made from polymers without cell seeding were killed and examined in a similar fashion after 8 weeks. In the control animals, the acellular polymer leaflets were completely degraded, with no residual leaflet tissue at 8 weeks. The tissue-engineered valve leaflet persisted in each animal in the experimental group. 4-Hydroxyproline analysis of the constructs showed a progressive increase in collagen content. Immunohistochemical staining demonstrated elastin fibers in the matrix and factor VIII on the surface of the leaflet. The cell-labeling experiments demonstrated that the cells on the leaflets had persisted from the in vitro seeding of the leaflets. CONCLUSIONS: In the tissue-engineered heart valve leaflet, transplanted autologous cells generated a proper matrix on the polymer scaffold in a physiological environment at a period of 8 weeks after implantation.

Animals↗
PubMed1996

Heparinase III from Flavobacterium heparinum: cloning and recombinant expression in Escherichia coli.

Heparinase III (E.C. 4.2.2.8), formerly heparinase I, produced by Flavobacterium heparinum is an enzyme that specifically cleaves heparan sulfate-rich regions of acidic polysaccharides. In this study, we report the cloning of the heparinase III gene using polymerase chain reaction (PCR). Two degenerate oligonucleotides, based on amino acid sequences derived from tryptic peptides of purified heparinase III were used to generate a approximately 1100-bp probe by PCR amplification using Flavobacterium genomic DNA as the template. The PCR-derived probe was used to screen a Flavobacterium genomic DNA library in lambda ZAP II. The open reading frame of the heparinase III gene is 1980 bp in length, encoding a precursor protein of 75,950 Da; 10 of the tryptic peptides mapped onto the open reading frame which corresponded to approximately 18% of the protein. Recombinant heparinase III was expressed in Escherichia coli using the T7 polymerase pET expression system. This is the first report of the cloning and recombinant expression of an enzyme primarily degrading heparan sulfate.

Amino Acid Sequence↗
PubMed1996

Heparinase I from Flavobacterium heparinum. Identification of a critical histidine residue essential for catalysis as probed by chemical modification and site-directed mutagenesis.

We recently identified cysteine-135 as an important amino acid for heparinase I (EC 4.2.2.7) activity. In this study, we have identified a second residue critical for enzymatic activity. We observe concentration-dependent inactivation of heparinase I in the presence of reversible histidine-modifying diethyl pyrocarbonate (DEPC); 0.3 mM DEPC results in 95% of heparinase I inactivation in less than 3 min, and as low as 10 microM DEPC results in a 85% loss of heparinase I activity in 15 min. Heparinase I activity is restored following hydroxylamine treatment. This, along with other experiments, strongly suggests that the inactivation of heparinase I by DEPC is specific for histidine residues. Chemical modification, under nondenaturing conditions, of the histidines using nonradiolabeled and [14C]DEPC indicates that between one and two histidine residues are modified. Chemical modification of the surface-accessible histidines, in the presence and absence of heparin, suggests that the histidine(s) lie(s) in or near the active site of heparinase I. The wild-type heparinase I has four histidine residues; site-directed mutagenesis of H129A, H165A, and H339A did not affect enzyme activity and the kinetic parameters, suggesting that these residues are not essential for heparinase I activity. However, H203A inactivates heparinase I while a H203D mutant has residual activity, indicating a role of this residue in catalysis. We propose that histidine-203, contained in the heparin binding site, is immediately adjacent to cysteine-135, and these residues together form the catalytic domain of heparinase I.

Amino Acid Sequence↗
PubMed1996

Expression in Escherichia coli, purification and characterization of heparinase I from Flavobacterium heparinum.

The use of heparin for extracorporeal therapies has been problematical due to haemorrhagic complications; as a consequence, heparinase I from Flavobacterium heparinum is used for the determination of plasma heparin and for elimination of heparin from circulation. Here we report the expression of recombinant heparinase I in Escherichia coli, purification to homogeneity and characterization of the purified enzyme. Heparinase I was expressed with an N-terminal histidine tag. The enzyme was insoluble and inactive, but could be refolded, and was purified to homogeneity by nickel-chelate chromatography. The cumulative yield was 43%, and the recovery of purified heparinase I was 14.4 mg/l of culture. The N-terminal sequence and the molecular mass as analysed by matrix-assisted laser desorption MS were consistent with predictions from the heparinase I gene structure. The reverse-phase HPLC profile of the tryptic digest, the Michaelis-Menten constant Km (47 micrograms/ml) and the specific activity (117 units/mg) of purified recombinant heparinase I were similar to those of the native enzyme. Degradation of heparin by heparinase I results in a characteristic product distribution, which is different from those obtained by degradation with heparinase II or III from F. heparinum. We developed a rapid anion-exchange HPLC method to separate the products of enzymic heparin degradation, using POROS perfusion chromatography media. Separation of characteristic di-, tetra- and hexa-saccharide products is performed in 10 min. These methods for the expression, purification and analysis of recombinant heparinase I may facilitate further development of heparinase I-based medical therapies as well as further investigation of the structures of heparin and heparan sulphate and their role in the extracellular matrix.

Amino Acid Sequence↗
PubMed1996

The nucleation of receptor-mediated endocytosis.

A theory of the mechanical origins of receptor-mediated endocytosis shows that a spontaneous membrane complex formation can provide the stimulus for a local membrane motion toward the cytosol. This motion is identified with a nucleation stage of receptor-mediated endocytosis. When membrane complexes cluster, membrane deformation is predicted to be most rapid. The rate of growth of membrane depressions depends upon the relative rates of approach of aqueous cytosolic and extracellular fluids toward the cell membrane. With cytosolic and extracellular media characterized by apparent viscosities, the rate of growth of membrane depressions is predicted to increase as the extracellular viscosity nears the apparent viscosity of the cytosol and then to decrease when the extracellular viscosity exceeds that of the cytosol. To determine whether these trends would be apparent in the overall endocytosis rate constant, an experimental study of transferrin-mediated endocytosis in two different cell lines was conducted. The experimental results reveal the same dependence of internalization rate on extracellular viscosity as predicted by the theory. These and other comparisons with experimental data suggest that the nucleation stage of receptor-mediated endocytosis is important in the overall endocytosis process.

Biophysical Phenomena↗
PubMed1996

Temporal study of the activity of matrix metalloproteinases and their endogenous inhibitors during wound healing.

The restoration of functional connective tissue is a major goal of the wound healing process. This regenerative event requires the deposition and accumulation of collagenous and noncollagenous matrix molecules as well as the remodelling of extracellular matrix (ECM) by matrix metalloproteinases (MMPs). In this study, we have utilized substrate gel electrophoresis, radiometric enzyme assays, and Western blot analyses to determine the temporal pattern of appearance and activity of active and latent MMPs and their inhibitors during the entire healing process in a partial thickness wound model. Through the use of substrate gel electrophoresis, we studied the appearance of proteolytic bands whose molecular weight was consistent with their being members of the MMP family of enzymes. Proteolytic bands whose molecular weight is consistent with both the active and latent forms of MMP-2 (72 kDa, Type IV gelatinase) were detected in wound fluid of days 1-7 after wounding. The number of active MMP-2 species detectable in wound fluid was greatest during days 4-6 after wounding. The most prominent proteolytic band detected each day migrated with a molecular weight consistent with it being the latent form of MMP-9 (92 kDa, Type V pro-collagenase). In contrast to MMP-2, the active form of this enzyme was never detected. The presence of MMP-1 (interstitial collagenase) was detected by immunoblot in the wound fluid from days 1-6 post-injury. Using a radiometric enzyme assay for collagenase inhibitory activity we have also determined the time course of activity of endogenous matrix metalloproteinase inhibitors. We have correlated these data to the known cellular events occurring in the wound during this time period as well. This study establishes a prototypical pattern of MMP appearance in normal wound healing. It may also provide potential intervention sites for the therapeutic use of inhibitors of aberrant MMP activities which characterize chronic wounds.

Animals↗
PubMed1996

Heparinase I from Flavobacterium heparinum. Mapping and characterization of the heparin binding domain.

In this study we have identified the primary heparin binding site of heparinase I (EC 4.2.2.7). Chemical and proteolytic digests of heparinase I were used in direct binding and competition assays, to map the regions of heparinase I that interact specifically with heparin. We find the heparin binding site contains two Cardin-Weintraub heparin binding consensus sequences and a calcium co-ordination consensus motif. We show that heparin binding to heparinase I is independent of calcium (Kd of 60 nm) and that calcium is able to activate heparinase I catalytically. We find that sulfhydryl selective labeling of cysteine 135 of heparinase I protects the lysines of the heparin binding sequence from proteolytic cleavage, suggesting the close proximity of the heparin binding site to the active site. Site-directed mutagenesis of H203A (contained in the heparin binding site) inactivated heparinase I; however, a H203D mutant retained marginal activity, indicating a role for this residue in catalysis. The above results taken together suggest that histidine 203 (hence the heparin binding site) is immediately adjacent to the scissile bond. We propose that the heparin binding site and active site are in close proximity to each other and that the calcium coordination motif, contained in the heparin binding site, may bridge heparin to heparinase I through calcium in a ternary complex during catalysis.

Amino Acid Sequence↗
PubMed1996

Preferential self-association of basic fibroblast growth factor is stabilized by heparin during receptor dimerization and activation.

Central to signaling by fibroblast growth factors (FGFs) is the oligomeric interaction of the growth factor and its high-affinity cell surface receptor, which is mediated by heparin-like polysaccharides. It has been proposed that the binding of heparin-like polysaccharides to FGF induces a conformational change in FGF, resulting in the formation of FGF dimers or oligomers, and this biologically active form is 'presented' to the FGF receptor for signal transduction. In this study, we show that monomeric basic FGF (FGF-2) preferentially self-associates and forms FGF-2 dimers and higher-order oligomers. As a consequence, FGF-2 monomers are oriented for binding to heparin-like polysaccharides. We also show that heparin-like polysaccharides can readily bind to self-associated FGF-2 without causing a conformational change in FGF-2 or disrupting the FGF-2 self-association, but that the bound polysaccharides only additionally stabilize the FGF-2 self-association. The preferential self-association corresponds to FGF-2 translations along two of the unit cell axes of the FGF-2 crystal structures. These two axes represent the two possible heparin binding directions, whereas the receptor binding sites are oriented along the third axis. Thus, we propose that preferential FGF-2 self-association, further stabilized by heparin, like "beads on a string," mediates FGF-2-induced receptor dimerization and activation. The observed FGF-2 self-association, modulated by heparin, not only provides a mechanism of growth factor activation but also represents a regulatory mechanism governing FGF-2 biological activity.

Apoproteins↗
PubMed1996

Imaging of fluorescent molecule and small ion transport through human stratum corneum during high voltage pulsing: localized transport regions are involved.

During the application of high-voltage pulses across the skin, transport of two negatively charged fluorescent molecules through the stratum corneum is highly localized. The apparent size of these localized transport regions (LTR's) is initially 10 microns in diameter for both calcein and sulforhodamine. Appearance of LTR's occurred at or above transdermal voltages of 75 V. In the range of 75 to 160 V, the number of LTR's increases with voltage, but their initial size is the same at all voltages; with additional pulses LTR's increase in size, reaching diameters of approximately 40-80 microns. Small ion currents across the skin are also localized and include the LTR's; however, the areas of current flux appeared to be larger. There was no visible damage to the structure of the skin seen at 100 x magnification for any of the voltages used (< 170 V across the skin). Significantly, LTR's are not sweat ducts or hair follicles.

Adult↗
PubMed1996

In vitro bone biocompatibility of poly (anhydride-co-imides) containing pyromellitylimidoalanine.

Poly(anhydride-co-imides) are currently under study for applications involving bone. The cytotoxicity of a series of poly(anhydride-co-imides) with osteoblast-like cells (MC3T3-E1) was evaluated. The imide component of the copolymers was based on pyromellitylimidoalanine and the anhydride component was based on either sebacic acid or 1,6-bis(carboxyphenoxy)hexane. Cell adhesion and proliferation on the surfaces of the polymer discs were observed by environmental scanning electron microscopy. During the first 24 hours of attachment, the cells showed normal morphology when cultured on copolymers containing 1,6-bis(carboxyphenoxy)hexane. The cells did not adhere to the polymers containing sebacic acid, probably due to the rapid degradation of the polymer surfaces. Concurrently, the effects of polymer breakdown products on osteoblast-like cells were evaluated by studying their proliferation (cell numbers), viability (dye exclusion), morphology (light microscopy), and phenotypic expression. The morphology of osteoblast-like cells cultured in the presence of the polymer breakdown products pyromellitylimidoalanine and pyromellitic acid was found to be similar to that of the same cells grown on tissue culture polystyrene and consisted of a characteristic polygonal shape. With use of a monoclonal antibody to osteocalcin, these cells were shown to demonstrate preserved osteoblast phenotype with growth over a 21-day period. In addition, the cells reached confluency after 3-4 days, similar to cells grown on tissue culture polystyrene. This in vitro evaluation showed that the poly(anhydride-co-imides) evaluated are non-cytotoxic and may be viable biomaterials for orthopaedic applications.

3T3 Cells↗