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Biomedical subjects

J Alvarez

Publications and source records attributed to J Alvarez.

At least 145 records · Page 8Linked to original sources
PubMed1996

Transient inhibition of capacitative calcium entry in human neutrophils by a monoclonal antibody directed against a 19-kDa antigen.

P1C3 is a monoclonal antibody that binds p19, a novel neutrophil activation antigen that translocates to the cell surface upon neutrophil activation. We find that P1C3 inhibits capacitative Ca2+ entry, induced by emptying the intracellular Ca2+ stores with thapsigargin. The effect is transient, reaching its maximum at 30-60 s, but becomes permanent upon pretreatment of the cells with the protein phosphatase inhibitor calyculin A, suggesting the involvement of protein phosphorylation. The inhibitory action is similar to the one reported previously for the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (fMLP), although the transduction mechanism may be different. Inhibition of Ca2+ entry by fMLP was prevented by pretreatment with pertussis toxin, whereas inhibition by P1C3 was not. Pretreatment with cholera toxin had no effect. This suggests that the effect of P1C3 may not be mediated by a heterotrimeric G protein. Tyrosine kinase inhibitors did not prevent inhibition by either fMLP or P1C3. Phospholipase C activation seems not to be involved as P1C3, contrarily to fMLP, was unable to induce Ca2+ release from the intracellular Ca2+ stores.

Antibodies, Monoclonal↗
PubMed1996

The development of blood transfusion: the contributions of Norman Bethune in the Spanish Civil War (1936-1939).

Although the Spanish Civil War (1936-1939) has been the focus of many papers in many countries, there are few references concerning medicine during this war, and many of the related events and medical progress are not well known. Some of these events came to influence civilian and military medicine. Among these were advances in blood transfusion, in which the Canadian surgeon, Norman Bethune, played an important role, particularly in advocating transfusion of blood to soldiers at the front rather than at medical centres behind the lines. While much of Bethune's work, especially in Canada and China, is well known, 1-3 certain aspects of Bethune's work and his stay in Spain are worth reviewing.

Blood Transfusion↗
PubMed1996

Axillary brachial plexus anesthesia: electrical versus cold saline stimulation.

The aim of this study was to investigate which of two methods of nerve stimulation, cold saline-induced paresthesia or use of a nerve stimulator, was more effective in increasing the successful brachial plexus block rate by the axillary approach. Twenty patients were randomly assigned to Group A (saline below 11 degrees C), and 20 patients to Group B (nerve stimulator). All blocks were performed by the same anesthesiologist using 40 mL of 1.5% mepivacaine and 4 mL of 8.4% sodium bicarbonate. Successful block was defined using Vester-Andersen et al.'s criteria. Cold saline-induced paresthesiae in the hand or forearm were obtained in 19 patients (95%) during one of four attempts allowed, and in 15 patients (75%) on the first attempt. A motor response was evoked by a nerve stimulator in 17 patients (85%). Two patients (10%) had a paresthesia in the hand without a motor response with the current at less than 1 mA. A successful block was achieved in 19 patients in each group.

Anesthetics, Local↗
PubMed1996

The N-glycan of the SCR 2 region is essential for membrane cofactor protein (CD46) to function as a measles virus receptor.

Membrane cofactor protein (MCP) (CD46), a complement-regulatory protein, serves as a cellular receptor for measles virus. Its amino-terminal portion is composed of four short consensus repeats (SCR), three of which (SCR1, SCR2, and SCR4) carry an N-linked oligosaccharide. In order to determine the importance of the three N-glycans for the function of MCP as a measles virus receptor, we established Chinese hamster ovary (CHO) cell lines that stably express mutant MCPs lacking one of the three motifs for N glycosylation (NQ1, NQ2, and NQ4). In an additional mutant (NQ1-2), two glycosylation motifs were altered, allowing the addition of an N-linked oligosaccharide only in SCR4. The abilities of the mutant MCPs to function as measles virus receptors were analyzed with three different assays: (i) binding of measles virus hemagglutinin to MCP immobilized on nitrocellulose; (ii) binding of measles virus to CHO cells expressing wild-type or mutant MCP; and (iii) infection of the transfected CHO cells by measles virus. In all three assays, the abilities of the NQ2 and NQ1-2 mutants to serve as measles virus receptors were drastically impaired. The NQ1 and NQ4 mutants were recognized by measles virus almost as efficiently as the wild-type protein. These results indicate that the N-glycan attached to SCR2 is essential for MCP to serve as a measles virus receptor, while the oligosaccharides attached to SCR1 and SCR4 are of only minor importance.

Animals↗
PubMed1996

[The discovery of surgical anesthesia and its arrival in Europe. Apropos of the 150th anniversary of the clinical introduction of ether].

The clinical introduction of anesthesia took place on the 16th of October 1846 at Massachusetts General Hospital (Boston) and William T.G. Morton, a dentist in the city, was its discoverer. The news was made public at the beginning of November and soon crossed the Atlantic, reaching Paris and London, where there were acclaimed medical centers. The event was the object of studies worldwide and still today receives the attention of researchers. We have detected numerous inaccuracies in most accounts of how anesthesia was introduced in Europe, motivating us to undertake the present study to establish a new history of the first uses of ether in European countries. We have consulted new bibliographic sources and obtained results that are considerably different from those published by most authors in recent years. Our analysis and discussion of the findings allow us to establish a new chronological account of anesthesia with ether in Europe, in which we emphasize the first studies of etherizations performed in Belgium, Spain and Italy hitherto ignored by other authors.

Anesthesia, Inhalation↗
PubMed1995

Monitoring dynamic changes in free Ca2+ concentration in the endoplasmic reticulum of intact cells.

Direct monitoring of the free Ca2+ concentration in the lumen of the endoplasmic reticulum (ER) is an important but still unsolved experimental problem. We have shown that a Ca(2+)-sensitive photoprotein, aequorin, can be addressed to defined subcellular compartments by adding the appropriate targeting sequences. By engineering a new aequorin chimera with reduced Ca2+ affinity, retained in the ER lumen via interaction of its N-terminus with the endogenous resident protein BiP, we show here that, after emptying the ER, Ca2+ is rapidly re-accumulated up to concentrations of > 100 microM, thus consuming most of the reporter photoprotein. An estimate of the steady-state Ca2+ concentration was obtained using Sr2+, a well-known Ca2+ surrogate which elicits a significantly slower rate of aequorin consumption. Under conditions in which the rate and extent of Sr2+ accumulation in the ER closely mimick those of Ca2+, the steady-state mean lumenal Sr2+ concentration ([Sr2+]er) was approximately 2 mM. Receptor stimulation causes, in a few seconds, a 3-fold decrease of the [Sr2+]er, whereas specific inhibition of the ER Ca2+ ATPase leads to an approximately 10-fold drop in a few minutes.

Aequorin↗
PubMed1995

Transfected aequorin in the measurement of cytosolic Ca2+ concentration ([Ca2+]c). A critical evaluation.

Targeted recombinant aequorins represent to date the most specific means of monitoring [Ca2+] in subcellular organelles (Rizzuto, R., Simpson, A. W. M., Brini, M., and Pozzan, T. (1992) Nature 358, 325-328; Brini, M., Murgia, M., Pasti, L., Picard, D., Pozzan, T., and Rizzuto, R. (1993) EMBO J. 12, 4813-4819; Kendall, J. M., Dormer, R. L., and Campbell, A. K. (1992) Biochem. Biophys. Res. Commun. 189, 1008-1016). Up until now, however, only limited attention has been paid to the use of recombinant photoproteins for measuring, in mammalian cells, the [Ca2+] in the cytoplasm, a compartment for which effective Ca2+ probes are already available. Here we describe this approach in detail, highlighting the advantages, under various experimental conditions, of using recombinant cytosolic aequorin (cytAEQ) instead of classical fluorescent indicators. We demonstrate that cytAEQ is expressed recombinantly at high levels in transiently transfected cell lines and primary cultures as well as in stably transfected clones, and we describe a simple algorithm for converting aequorin luminescence data into [Ca2+] values. We show that although fluorescent indicators at the usual intracellular concentrations (50-100 microM) are associated with a significant buffering of the [Ca2+]c transients, this problem is negligible with recombinantly expressed aequorin. The large dynamic range of the photoprotein also allows an accurate estimate of the large [Ca2+]c increases that are observed in some cell types such as neurons. Finally, cytAEQ appears to be an invaluable tool for measuring [Ca2+]c in cotransfection experiments. In particular, we show that when cotransfected with an alpha 1-adrenergic receptor (coupled to inositol 1,4,5-trisphosphate generation), cytAEQ faithfully monitors the subpopulation of cells expressing the receptor, whereas the signal of fura-2, at the population level, is dominated largely by that of the untransfected cells.

Aequorin↗
PubMed1995

Incorporation of amino acids into the axoplasm is enhanced by electrical stimulation of the fiber.

The effect of sustained electrical stimulation upon the incorporation of amino acids into the axoplasm was studied in the goldfish Mauthner (M) axon with light autoradiography. An extracellular pulse of tracers applied between M-axons in the medulla resulted in a local and substantial labeling of the M-axoplasm and a faint labeling of the M-perikaryon 4-5 mm away from the site of injection. After 18 h of direct electrical stimulation of the M-axon at 0.3-0.8 Hz, the local incorporation of amino acids into the M-axoplasm doubled. This enhancement declined to reach the baseline within 24 h. A 4 h electrical stimulation did not enhance the incorporation. Transynaptic activation of the M-neuron through the auditory input at 0.1-0.2 Hz for 18 h did not raise the amino acid incorporation in the M-axoplasm. We conclude that electrical discharge of the axon modulates the local incorporation of amino acids into the axoplasm.

Amino Acids↗
PubMed1995

Measles virus and C3 binding sites are distinct on membrane cofactor protein (CD46).

The human complement regulatory protein membrane cofactor protein (CD46) is the cellular receptor for measles virus (MV), whereas decay accelerating factor (DAF; CD55), a structurally similar complement regulatory protein, does not bind MV. To characterize the interaction between MV and CD46, mutants of the CD46 protein and hybrid molecules between CD46 and DAF were tested for their ability to act as MV receptors. The transmembrane domain and cytoplasmic tail of CD46 were not required for receptor function as cells expressing the CD46 extracellular domain linked to the glycosyl-phosphatidylinositol tail of DAF were rendered susceptible to MV infection. Chimeric proteins exchanging the four extracellular short consensus repeat (SCR) domains between CD46 and DAF indicated that only molecules with both SCR1 and SCR2 from CD46 allowed a productive MV infection. Further, monoclonal antibodies (mAbs) against SCR1 or SCR2 of CD46 blocked MV infection, whereas a mAb against SCR3 and SCR4 did not. The latter mAb blocks C3b/C4b binding (which maps to SCR3 and SCR4) whereas the former mAbs do not. Thus, our data indicate that both SCR1 and SCR2 make up the MV receptor determinant in CD46. These results also suggest avenues for development of therapeutic agents to inhibit MV binding and thus infection and disease.

Animals↗
PubMed1995

Biphasic and differential modulation of Ca2+ entry by ATP and UTP in promyelocytic leukaemia HL60 cells.

ATP and UTP cause mobilization of Ca2+ from the intracellular stores with similar potency in several cell types including both undifferentiated and differentiated HL60 cells. We show here that, in HL60 cells with Ca2+ stores that had been fully and irreversibly emptied using the endomembrane Ca(2+)-ATPase inhibitor thapsigargin, both nucleotides produced a biphasic effect on Ca2+ entry, first rapid inhibition and then delayed (about 15 s) activation. ATP was more effective at producing the initial inhibition of Ca2+ entry, whereas UTP was more effective at activating the delayed Ca2+ entry. Previous incubation with UTP desensitized the Ca2+ mobilization and the delayed activation of Ca2+ entry induced by ATP but not the inhibition of Ca2+ entry. The ATP analogue 2-methylthioATP (2-MeSATP) barely mobilized stored Ca2+ but inhibited Ca2+ entry. These results could be explained by the presence of two receptors: (i) a P2u receptor sensitive to ATP and UTP, responsible for activation of phospholipase C and Ca2+ mobilization, early inhibition of Ca2+ entry and delayed activation of Ca2+ entry and (ii) a P2y-like receptor sensitive to ATP and 2-MeSATP which produces only inhibition of Ca2+ entry. The inhibition of Ca2+ entry by nucleotides increased greatly during differentiation. Given that Ca2+ mobilization by nucleotides is not modified by differentiation, this suggests that a component of the mechanism of inhibition of Ca2+ entry is gradually expressed during differentiation of HL60 cells.

Adenosine Triphosphate↗
PubMed1995

Mitosis of Schwann cells and demyelination are induced by the amyloid precursor protein and other protease inhibitors in the rat sciatic nerve.

We studied the cytological alterations produced in the rat sciatic nerve by the amyloid precursor protein (APP) containing the Kunitz insert (APP K+) and other protease inhibitors. Conditioning of nerve segments with APP K+, aprotinin or leupeptin for 5 days or more resulted in mitosis of Schwann cells, demyelination of fibres, and a < 10-fold increase in Schwann cells, associated with demyelinated fibres. Altered fibres nevertheless involved a small part of the population. Nerve segments proximal and distal to the conditioned region showed almost no alteration. Conditioning with saline, heated APP K+, or APP without the Kunitz insert was not effective. We conclude that APP K+ and other protease inhibitors induce Schwann cells to enter the cell cycle, and once committed to proliferate they resorb their myelin. These functional properties of APP may be relevant to the pathogenesis of Alzheimer's disease.

Amyloid beta-Protein Precursor↗
PubMed1995

Anaerobic threshold in children: determination from saliva analysis in field tests.

The purpose of this study was to determine the anaerobic threshold of children by the analysis of saliva collected during field tests. A group of 25 children (mean age, 10.5 years) performed an incremental exercise test on a track, consisting of 4-min stages at increasing running velocities. Before each test (at rest) and at the end of each stage, both blood (via finger pricks) and saliva samples (for measurement of salivary concentrations of Na+ and Cl-) were collected to determine lactate threshold (Thla-) and saliva threshold (Thsa), respectively. There were no significant differences between values of Thla- and Thsa when expressed either as running velocity [mean Thla-, 10.73 (SD 1.96) km.h-1; mean Thsa, 10.89 (SD 1.69) km.h-1)] or heart rate [Thla-, 182(SD 14) beats. min-1 Thsa 183 (SD 11) beats.min-1]. In addition, correlations between Thsa and Thla were high, when both values were expressed as running velocity in kilometres per hour (r = 0.89; P < 0.001), or heart rate in beats per minute (r = 0.90; p < 0.001). In conclusion, these findings suggested that saliva analysis would be a valid method for anaerobic threshold determination in field tests.

Anaerobic Threshold↗
PubMed1995

Early axonal regeneration: repression by Schwann cells and a protease?

We have proposed that mature Schwann cells and an extracellular protease repress the sprouting response of axons. To test this hypothesis, we destroyed all cells by freezing a short span of the rat sciatic nerve or inhibited proteases with subperineurial injections of aprotinin, and a crush was made to induce the sprouting response. In unconditioned or in vehicle-injected nerves, axons began to elongate at a constant rate after a delay of about 1 day. The freezing of the nerve distal to the crush obliterated the delay, but the rate of elongation did not change. A similar pattern was observed when the nerve segment was conditioned with aprotinin for 2 days prior to the crush. These effects were abolished when a short untreated segment was left between crush and conditioned region of the nerve. The electron microscopy of the nerve and the immunolocalization of the growth-associated protein (GAP-43) were consistent with the enhanced regrowth observed in conditioned nerves. Our findings support the notion that Schwann cells repress the onset of regeneration and that a local protease is involved.

Animals↗
PubMed1995

Bradycardia and asystole following atropine-neostigmine administration after caesarean section in a parturient receiving methyldopa for pregnancy-induced hypertension.

We report one case of bradycardia and asystole immediately after the administration of 1 mg atropine and 2 mg neostigmine after the completion of an urgent caesarean section. We attribute this adverse reaction to the treatment of pregnancy-induced hypertension with methyldopa, perhaps facilitated by other drugs employed. Similar reactions have been reported relating to beta-receptor antagonists and tricyclic antidepressants, but not to methyldopa.

Journal Article↗