Self-recognition by proteoglycans.
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Animals↗
Biomedical subjects
O Popescu
Publications and source records attributed to O Popescu.
[Treatment of acute stroke on the stroke unit. Initial experiences with an acute stroke unit in Germany].
We studied the effects of stroke unit care in an acute Neurology department on the outcome and the length of hospital stay in acute stroke patients. After an emergency evaluation on admission including CCT, ultrasound studies and ECG the patients were treated on a specialist stroke unit for an average 3.9 days. For 48 +/- 24 h monitoring of blood pressure, ECG, pO2 and transcranial Doppler sonography was instituted. Stroke unit treatment reduced the length of hospital stay from an average 15.8 days in the time period before institution of the stroke unit to 12.9 days. This effect was especially marked in patients with acute occlusion of major intracranial arteries (before stroke unit treatment: n = 33; hospital stay 22.5 days; after stroke unit treatment: n = 54; hospital stay 13.9 days). Clinical deterioration in acute ischemic stroke was related to reduction of cerebral blood flow velocities due to blood pressure changes or space occupying effects. Monitoring on the stroke unit allowed immediate treatment of systemic hypotension, cerebral edema or cardiac arrhythmias. Transcranial Doppler sonography revealed HITS in 6/55 acute stroke patients without new clinical symptoms. Monitoring on the stroke unit improved the specific care for acute stroke patients. The length of hospital stay was reduced after stroke unit care.
Acute Disease↗
Expression and biological activity of genetic fusions between MalE, the maltose binding protein from Escherichia coli and portions of CD4, the T-cell receptor of the AIDS virus.
Hybrid molecules between MalE, the periplasmic maltose binding protein of Escherichia coli, and CD4, the human T-lymphocyte receptor for the AIDS virus HIV, have been constructed and purified. We show that CD4 can be fused as multiple repeats to both ends of a single MalE molecule. Hybrid proteins are exported into the periplasm of bacteria, bind monoclonal antibodies directed against CD4, bind HIV gp160, and inhibit HIV binding to CD4+ cells. MalE has been used as a scaffold to graft portions of CD4. Deletion analysis allowed to define a minimal structural domain which folds in a way which is compatible with its biological activity. This minimal part was used to design compact hybrid molecules in which CD4 was inserted internally into MalE.
ATP-Binding Cassette Transporters↗
Disseminated cerebral hemorrhages as unusual manifestation of toxoplasmic encephalitis in AIDS.
Hemorrhagic lesions are uncommon in central nervous system (CNS) toxoplasmosis. We report here the case of a patient with multiple cerebral hemorrhages as the initial manifestation of HIV infection due to CNS toxoplasmosis.
AIDS-Related Opportunistic Infections↗
Assessment of the posterior communicating artery by transcranial color-coded duplex sonography.
BACKGROUND AND PURPOSE: The aim of this study was to investigate flow velocity and flow direction in the posterior communicating artery (PcomA) by means of transcranial color-coded duplex sonography (TCCD) and to compare the results with angiographic findings. METHODS: Thirty patients with unilateral occlusion of the internal carotid artery (ICA) due to atherosclerosis (n=15) or balloon occlusion (n=15) and 50 normal subjects were included. The circle of Willis was insonated through the temporal bone window. In 24 patients with unilateral ICA occlusion, angiograms were available and were compared with the results of TCCD. RESULTS: The PcomA could be detected unilaterally in 70% of normal subjects and bilaterally in 30%. A retrograde flow direction in the PcomA from the posterior cerebral artery to the ICA was found in 75% of the normal control subjects. The mean peak flow velocity in normal PcomAs was 36 +/- 15 cm/s (+/- SD). No significant differences in flow velocity were found between unilaterally and bilaterally detectable PcomAs or between retrograde and orthograde PcomAs. In patients with unilateral ICA occlusion we observed ipsilaterally a retrograde flow direction, with an elevation of flow velocity (64 +/- 10 cm(s) compared with the contralateral side (27 +/- 14 cm/s; P<.001). CONCLUSIONS: TCCD appears to be a valuable method to determine flow velocity and flow direction not only in the large intracranial vessels but also in the smaller communicating arteries. In the future this method could be useful for the planning of ICA balloon occlusions and in deciding whether to perform extracranial/intracranial bypass surgery. It could furthermore show intracranial collaterals in patients with cerebrovascular disease and help to estimate the risk of watershed infarctions in patients with asymptomatic high-grade ICA stenosis and in patients undergoing carotid endarterectomy.
Adult↗
Binding strength between cell adhesion proteoglycans measured by atomic force microscopy.
Measurement of binding forces intrinsic to adhesion molecules is necessary to assess their contribution to the maintenance of the anatomical integrity of multicellular organisms. Atomic force microscopy was used to measure the binding strength between cell adhesion proteoglycans from a marine sponge. Under physiological conditions, the adhesive force between two cell adhesion molecules was found to be up to 400 piconewtons. Thus a single pair of molecules could hold the weight of 1600 cells. High intermolecular binding forces are likely to form the basis for the integrity of the multicellular sponge organism.
Animals↗
Sulfate restriction induces hyposecretion of the adhesion proteoglycan and cell hypomotility associated with increased 35SO4(2-) uptake and expression of a band 3 like protein in the marine sponge, Microciona prolifera.
Sulfate is an important component relating to normal proteoglycan secretion and normal motility in the marine sponge, Microciona prolifera. The following alterations were observed in sponge cells when sulfate free artificial sea water was used as the suspension medium: 1) impairment of aggregation, 2) loss of cell movements, 3) a marked reduction in the secretion of the adhesion proteoglycan (AP). Reversal of this effect occurred if sulfate depleted cells were again rotated in sulfate containing artificial sea water. Motility and reaggregation of sulfate deprived cells could be completely restored by purified AP, but only if cells were first pre-conditioned in normal sea water. Comparisons of 35SO4(2-) uptake between normal and sulfate deprived cells which had been treated to reduce preformed secretions showed a marked increase in 35SO4(2-) uptake and incorporation which could be greatly augmented in the presence of Ca2+/Mg2+. Excessive retention of AP in sulfate starved cells demonstrated by immunostaining suggested that AP secretion and cellular motility may be controlled by a sulfate dependent secretogogue or that undersulfated AP itself had developed a secretory defect. SDS-PAGE of Triton treated cellular extracts demonstrated a 116 kDa 35SO4(2-) sulfated band which co-migrated with AP, but only in extracts derived from sulfate starved cells. Western blots prepared from such extracts incubated in the presence of a monoclonal anti-band 3 antibody demonstrated labelling of a single 97 kDa band only in material from sulfate deprived cells. The absence of this component in normal cell extracts indicated that this protein may be involved in facilitated sulfate transport. This study lends support to a heretofore unrecognized role for sulfate in cell motility and secretion.
Animals↗
The two soluble forms of the lipopolysaccharide receptor, CD14: characterization and release by normal human monocytes.
CD14, a glycolipid-anchored membrane glycoprotein, acts as a high affinity lipopolysaccharide receptor on leukocytes. We previously reported that the Mono-Mac-6 cell line releases two different soluble forms of CD14 (sCD14) (Labeta et al., Eur. J. Immunol. 1993. 23: 2144). Here we show that the two sCD14, which we now refer to as sCD14 alpha (low M(r)) and sCD14 beta (high M(r)), are also synthesized and released by normal human monocytes and present in normal plasma. Their mechanism of release was examined by using the Mono-Mac-6 cell line, chinese hamster ovary cell (CHO)/CD14+ transfectants and plasma from paroxysmal nocturnal hemoglobinuria (PNH) patients. It was found that: (1) sCD14 beta is released faster than sCD14 alpha and that the release of the latter is a lengthy process. (2) Monensin blocked the biosynthesis of membrane-bound CD14 (mCD14) and sCD14, additionally, a 50-kDa CD14 polypeptide accumulated in the cell lysate, suggesting that the different forms of CD14 may have a common precursor. (3) Monensin also blocked the release of sCD14 alpha from surface-labeled cells, suggesting that conversion of mCD14 to sCD14 alpha involves a mechanism of endocytosis followed by exocytosis. Interestingly, (4) sCD14 alpha and sCD14 beta were detected in PNH plasma, indicating that sCD14 alpha may also derive from an endogenous pathway. (5) Phospholipase C-released CD14 was identical in size to mCD14, thus differed from sCD14 beta by approximately 2000, indicating that release of sCD14 beta involves further processing. (6) CHO cells transfected with a CD14 cDNA coding for an eight C-terminal amino acids shorter product released an sCD14 beta-like form; thus absence of the eight C-terminal amino acids prevented mCD14 expression but not the secretion of sCD14 beta. The characterization of sCD14 alpha and sCD14 beta reported here may be useful for better understanding of variations in sCD14 levels in pathological conditions and the contribution of each sCD14 in sepsis and other, as yet unknown functions.
Animals↗
Comparative nuclear magnetic resonance studies of diffusional water permeability of red blood cells from sheep and cow.
1. The diffusional water permeability (p) of sheep and cow red blood cell (RBC) membrane has been monitored by a doping nuclear magnetic resonance (NMR) technique on control cells and following inhibition with p-chloromercuribenzene sulfonate (PCMBS). 2. There were no significant differences in the water permeability of sheep and cow RBCs, the values of p being around 3 x 10(-3) cm/sec at 20 degrees C and 5 x 10(-3) cm/sec at 37 degrees C. 3. Systematic studies of the effects of PCMBS on water diffusion indicated that in both species the maximal inhibition is reached in 60-90 min at 37 degrees C with 1 mM PCMBS. 4. The degree of inhibition increased as the temperature of measurement decreased, regardless of PCMBS concentration and incubation time. 5. The values of maximal inhibition ranged from 60-70% at 20 degrees C to 50-60% at 37 degrees C in the case of sheep RBCs, and from 45-55% at 20 degrees C to 40-50% at 37 degrees C in the case of cow RBCs. 6. The basal permeability to water of sheep RBCs was estimated as 1.0 x 10(-3) cm/sec at 20 degrees C and 2.2 x 10(-3) cm/sec at 37 degrees C, and that of cow RBCs as 1.6 x 10(-3) cm/sec at 20 degrees C and 2.7 x 10(-3) cm/sec at 37 degrees C. 7. In both species the activation energy of water diffusion was around 23 kJ/mol in control cells and reached values of around 30 kJ/mol after incubation with PCMBS in conditions of maximal inhibition of water diffusion.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Molecular and cellular targeting in the expression of foreign polypeptides in bacteria.
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ATP-Binding Cassette Transporters↗
Effects on water diffusion of inhibitors affecting various transport processes in human red blood cells.
The water permeability of human red blood cells has been monitored by nuclear magnetic resonance (NMR) following exposure to inhibitors of various transport processes across their membranes. No significant inhibition of water diffusion could be detected after the treatment of red blood cells with the anion exchange transport inhibitor dihydro-4,4'-diisothiocyano-stilbene-2,2'-disulfonate (H2DIDS) or the glucose transport inhibitors diallyl-diethyl-stilbestrol (DADES), cytochalasin B, or 30 mM iodoacetamide. It is for the first time that the effects of glucose transport inhibitors has been studied in detail by the NMR approach. A special case proved to be phloretin, an inhibitor of anion, nonelectrolyte and glucose permeability. A small but statistically significant inhibition of water permeability (around 12% at 20 degrees C) was induced by exposure to 2 mM phloretin (for 60 min at 37 degrees C); after a pretreatment of cells with 12 mM N-ethylmaleimide (NEM), for 60 min at 37 degrees C, the degree of inhibition induced by phloretin increased (becoming 17% at 20 degrees C). None of the inhibitors prevented or potentiated the strong inhibitory effect on water diffusion of a mercurial, p-chloromercuribenzene sulfonate (PCMBS). No increase in the activation energy of water diffusion occurred by treatment with the reagents used (exception the effect of PCMBS). The present results clarify some conflicting reports concerning the effects on water permeability of inhibitors of various transport processes in red blood cells and indicate that in addition to the drastic inhibition induced by mercurials other reagents may also have inhibitory effects.
4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
Water transport in human red cells: effects of 'non-inhibitory' sulfhydryl reagents.
The water diffusional permeability of human red blood cells following exposure to various sulfhydryl group (SH) reagents have been studied using a nuclear magnetic resonance technique. Exposure of red blood cells up to 12 mM N-ethylmaleimide (NEM) or 10 mM 5,5'-dithio-bis(2-nitrobenzoic acid) (DTNE) alone does not affect water diffusion. In contrast, when DTNB treatment follows a preincubation of the cells with NEM, a small (18% at 37 degrees C) but significant inhibition of water permeability occurs. The NEM and DTNB treatment of the cells caused no change of the cell shape and volume or of the cell water volume. Consequently, the inhibition observed after NEM and DTNB treatment has a real significance.
Cell Membrane Permeability↗
Amino acid composition of band 3 protein from red blood cells of normal and epileptic children.
Amino acid analyses of the band 3 protein purified from erythrocyte membranes of control and epileptic children showed that no major structural abnormalities of this protein could be linked with the red blood cell membrane alterations previously described in child epilepsy and, consequently, the molecular basis of these alterations should be looked for elsewhere.
Adolescent↗
The basal permeability to water of human red blood cells evaluated by a nuclear magnetic resonance technique.
The characteristics of water diffusional permeability (P) of human red blood cells were studied on isolated erythrocytes by a doping nuclear magnetic resonance technique. In order to estimate the basal permeability the maximal inhibition of water diffusion was induced by exposure of red blood cells to p-chloromercuribenzene sulfonate (PCMBS) under various conditions (concentration, duration, temperature). The lowest values of P were around 0.7 X 10(-3) cm s-1 at 10 degrees C, 1.2 X 10(-3) cm s-1 at 15 degrees C, 1.4 X 10(-3) cm s-1 at 20 degrees C, 1.8 X 10(-3) cm s-1 at 25 degrees C, 2.1 X 10(-3) cm s-1 at 30 degrees C and 3.5 X 10(-3) cm s-1 at 37 degrees C. The mean value of the activation energy of water diffusion (Ea,d) was 25 kJ/mol for control and 43.7 kJ/mol for PCMBS--inhibited erythrocytes. The values of P and Ea,d obtained after induction of maximal inhibition of water diffusion by PCMBS can be taken as references for the basal permeability to water of the human red blood cell membrane.
4-Chloromercuribenzenesulfonate↗
Studies of water permeability and proteins of erythrocyte membranes in patients with Duchenne muscular dystrophy.
The characteristics of water permeability of erythrocytes from 54 Duchenne muscular dystrophy (DMD) patients and age-matched controls have been determined by a pulse nuclear magnetic resonance (NMR) technique. A decreased permeability of erythrocyte membrane in DMD was definitely found at all temperatures between 15 and 42 degrees C, with normal values for the activation energy of water diffusion. No differences between DMD and control subjects in the pattern of erythrocyte membrane polypeptides separated by two-dimensional electrophoresis could be detected. The findings are discussed in relation to the molecular mechanisms of water diffusion across erythrocyte membrane and the problem of erythrocyte membrane abnormalities in DMD. A new interpretation of erythrocyte membrane alterations is proposed based on the recent findings regarding the molecular pathology of DMD.
Adolescent↗
Water exchange through erythrocyte membranes: biochemical and nuclear magnetic resonance studies re-evaluating the effects of sulfhydryl reagents and of proteolytic enzymes on human membranes.
The water permeability of human red blood cell (RBC) membrane has been monitored by a doping nuclear magnetic resonance (NMR) technique on intact cells and resealed ghosts following exposure to various sulfhydryl-reacting (SH) reagents and proteolytic enzymes. The main conclusions are the following: (i) When appropriate conditions for exposure of erythrocytes or ghosts to mercury-containing SH reagents (concentration, temperature and duration of incubation) were found, the maximal inhibition of water diffusion could be obtained with all mercurials (including HgCl2 and mersalyl that failed to show their inhibitory action on RBC water permeability in some investigations). While previous studies claimed that long incubation times are required for the development of maximal inhibition of water diffusion by mercurials, the present results show that it can be induced in a much shorter time (5-15 min at 37 degrees C) if relatively high concentrations of PCMBS (2-4 mM) are used and no washings of the inhibitor are performed after incubation. Higher than optimal concentrations of mercurials and/or longer incubation times result in lower values of inhibition, sometimes a loss of inhibition, or can even lead to higher values of permeability compared to control RBCs. (ii) The conditions for inhibition by mercurials are drastically changed by preincubation of erythrocytes with noninhibitory SH reagents (such as NEM or IAM) or by exposure to proteolytic enzymes. If the cells are digested with papain, the duration of incubation with PCMBS should be decreased in order for inhibition to occur. This explains the lack of inhibition reported previously, when a relatively long duration of incubation with PCMBS was used subsequent to papain digestion.(ABSTRACT TRUNCATED AT 250 WORDS)
Adolescent↗
Effects of temperature on water diffusion in human erythrocytes and ghosts--nuclear magnetic resonance studies.
The temperature-dependence of water diffusion across human erythrocyte membrane was studied on isolated erythrocytes and resealed ghosts by a doping nuclear magnetic resonance technique. The conclusions are the following: (1) The storage of suspended erythrocytes at 2 degrees C up to 24 h or at 37 degrees C for 30 min did not change the water exchange time significantly, even if Mn2+ was present in the medium. This indicates that no significant penetration of Mn2+ is taking place under such conditions. (2) In case of cells previously incubated at 37 degrees C for longer than 30 min with concentrations of p-chloromercuribenzene sulfonate (PCMBS) greater than 0.5 mM, the water-exchange time gradually decreased if the cells were stored in the presence of Mn2+ for more than 10 min at 37 degrees C. (3) When the Arrhenius plot of the water-exchange time was calculated on the basis of measurements performed in such a way as to avoid a prolonged exposure of erythrocytes to Mn2+ no discontinuity occurred, regardless of the treatment with PCMBS. (4) No significant differences between erythrocytes and resealed ghosts regarding their permeability and the activation energy of water diffusion (Ea,d) were noticed. The mean value of Ea,d obtained on erythrocytes from 35 donors was 24.5 kJ/mol. (5) The value of Ea,d increased after treatment with PCMBS, in parallel with the percentage inhibition of water diffusion. A mean value of 41.3 kJ/mol was obtained for Ea,d of erythrocytes incubated with 1 mM PCMBS for 60 min at 37 degrees C and 28.3 kJ/mol for ghosts incubated with 0.1 mM PCMBS for 15 min, the values of inhibition being 46% and 21% respectively.
4-Chloromercuribenzenesulfonate↗
[Progressive multifocal leukoencephalopathy. Late complication in chronic lymphatic leukemia].
In a 44 year old patient with chronic lymphatic leukemia and secondary antibody deficiency syndrome, a disorder of articulation and a left hemiparesis developed during a thrombocytopenic phase. Computer tomography of the cranium, CSF diagnostics and the electroencephalogram did not provide any indication for the cause of the rapidly progressive cerebral symptoms. In the NMR tomogram, a diffusedly increasing intensity in the T2-weighted tomograms were shown around the central region in the right brain. The patient died of a Pseudomonas septicemia. At autopsy, a typical finding of progressive multifocal leukoencephalopathy was found in the areas altered in the NMR tomography. Papova-like virions could be demonstrated in glial cells by electron microscopy.
Adult↗