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Biomedical subjects

C Márquez

Publications and source records attributed to C Márquez.

At least 19 recordsLinked to original sources

B cell triggering by bacterial lipopeptide involves both translocation and activation of the membrane-bound form of protein kinase C.

B cell activation by the lipopeptide trispalmitoyl-cysteinyl-alanyl-glycine (TPP), the biologically active moiety of bacterial lipoprotein, results in protein kinase C (PKC) translocation from the cytosol to the plasma membrane, as well as a significant increase in the activity of membrane-associated PKC that can be observed by in vitro incubation with TPP of partially purified PKC at calcium concentrations in the range of those prevailing in unstimulated B cells. TPP does not affect either the phosphoinositide turnover or the cytosolic-free calcium concentration, but promotes an increase in the intracellular pH that can be blocked by the PKC-inhibitors staurosporine or H-7. Moreover, incubation of B cells with staurosporine suppressed the proliferative response promoted by TPP at a half-maximum effective dose of 16 nM. Activation by TPP of PKC isoenzymes resolved after hydroxylapatite chromatography revealed that the resulting beta I/beta II isoenzyme was more sensitive than the alpha isoenzyme. These results suggest that TPP might mediate B cell activation via interaction with the membrane-associated fraction of PKC.

Animals

Protein kinase C mobilization in B lymphocytes. Differential isoenzyme translocation upon activation.

The subcellular distribution of protein kinase C has been analyzed in murine B lymphocytes exposed to LPS, anti-IgM antibodies and phorbol dibutyrate. An accurate determination of the enzyme mobilized from the soluble to the particulate fractions by these activators, has been made possible by the use of B cells in which the major part of the activity was present in the cytosol. Upon stimulation, we have analyzed the isoenzymatic forms translocated to the B cell membrane, showing a differential pattern of isoenzyme mobilization between LPS and anti-IgM antibodies. These data, together with the different Ca2+ requirements for the activation of the translocated protein kinase C isoenzymes, might help to unravel the mechanism responsible for the clonal expansion and differentiation of B lymphocytes, induced by the two ligands.

Animals

[Neuralgiform paroxysmal migraine].

A 23-year-old woman presented with episodic, strictly unilateral, left-sided headaches of excruciating severity. Pain was referred to the eye, lasted from seconds to a minute and was accompanied by homolateral ptosis, redness of the eye, increased lacrimation and nasal discharge. The attacks of pain repeated up to seven times over 24 hours and clustered around ovulation for seven days a month. After she became pregnant, the attacks increased in frequency and appeared every five to ten minutes during night and day. Different medical treatments including indomethacin, were without effect. Two months later pregnancy was interrupted and the pain immediately subsided. After carotid angiography the pain reappeared for two months, but it finally disappeared and she has been free of pain without treatment for the last nine months. This syndrome can be related to episodic paroxysmal hemicrania.

Abortion, Induced

[Complex I (NADH coenzyme-Q-reductase) deficiency, MELAS syndrome and hypertrophic cardiomyopathy].

A 24-year-old male had a deficiency of the complex I (NADH coenzyme-Q-reductase) of the mitochondrial respiratory chain, which clinically presented as a mitochondrial encephalomyopathy, with lactic acidosis and stroke-like episodes (MELAS syndrome). The encephalopathic episodes were preceded by migraine and were characterized by focal deficit signs, motor partial seizures and hypodense areas in the CT scan. An echocardiographic diagnosis of hypertrophic cardiomyopathy without intracavitary thrombi was made. It is suggested that hypertrophic cardiomyopathy is caused by the mitochondrial abnormalities that have been reported in the myocardium, and that migraine and cerebral infarctions are associated with abnormalities in the mitochondria from the endothelium and smooth muscle fibres of the cerebral small arteries and arterioles.

Acidosis, Lactic

Identity of PB76 differentiation antigen and lymphocyte alkaline phosphatase.

Alkaline phosphatases (APases, EC 3.1.3.1) are ecto-enzymes bound to cell membranes by a phosphatidyl-inositol anchor. We have previously shown that APase is present on activated murine B cells and its expression correlates with the process of B cell differentiation into immunoglobulin secretion. Recently, a monoclonal antibody (mAb), G-5-2, that recognizes a 76-kDa molecule preferentially expressed on the surface of pre-B and plasma cells (PB76) was described. Some features shared by APase and PB76 differentiation antigen suggest that the G-5-2 mAb might be specific for lymphocyte APase. Here, we have analyzed this possibility and found an absolute correlation between PB76 expression in cells and their APase activity. Although PB76 has been described as a B cell-restricted marker, PB76 is also expressed on some T cells, such as the YAC-1 T cell lymphoma, that are known to bear APase. Treatment of YAC-1 cells with phosphatidylinositol-specific phospholipase C resulted in a quantitatively correlated removal of both APase and PB76 antigens. Moreover, we demonstrate that PB76 antigen has APase activity using an enzyme-antigen immunoassay with the G-5-2 mAb. We conclude that PB76 and lymphocyte APase are one and the same antigen.

Alkaline Phosphatase

Activation of the interleukin 2 pathway precedes CD3-T cell receptor expression in thymic development. Differential growth requirements of early and mature intrathymic subpopulations.

T cell activation induced via the CD3-T cell receptor (TcR) complex, or by triggering of polyclonal antigen-independent pathways, involves both interleukin 2 (IL 2) production and IL 2 receptor (IL 2R) expression and results in T cell proliferation. To assess the potential role of the IL 2 pathway in T cell development, growth and activation requirements of intrathymic T cell precursors were analyzed and correlated with the expression of IL 2R. In contrast to CD3-TcR+ (either CD4+ or CD8+) mature thymic cells, CD3-TcR- CD1-4-8- early prothymocytes constitutively expressed IL 2R and displayed IL 2-mediated proliferation, which was inhibited by anti-IL 2R monoclonal antibodies (mAb). Moreover, prothymocytes developed spontaneous proliferation in the absence of exogenous IL 2, which was also abrogated by blocking of IL 2R with specific mAb. The possibility that both IL 2 production and IL 2R expression are autonomously activated early in T cell development, before acquisition of the CD3-TcR complex, led us to study the implication of alternative pathways of activation at this ontogenic stage. Triggering of the CD2 pathway with mAb against two different epitopes of the molecule (D66 and 9.6/T11(1) induced proliferation of CD3-TcR- prothymocytes in the absence of exogenous IL 2, while proliferation of CD3-TcR+ mature thymocytes required Il 2 supplementation. These data suggest that polyclonal activation of the Il 2 pathway may be selectively operative at early stages of T cell development, being involved in the growth and differentiation of intrathymic T cell precursors.

Antigens, Differentiation, T-Lymphocyte

Lack of correlation between translocation and biological effects mediated by protein kinase C: an appraisal.

Protein kinase C is involved in the mechanism of action of hormones, growth factors, mitogens and tumour promoters. The correlation between the extent of the biological effects mediated by protein kinase C and its fractional activation shows cell type specific patterns of behaviour. The discrepancy between enzyme activity and biological effects elicited by protein kinase C is particularly relevant to lymphoblastic cells. In B cells, the full expression of some biological responses mediated by protein kinase C may be achieved by activation of less than 5% of the enzyme activity.

Animals

[The state of muscle cramp disease].

A 65-year-old man presented with daily, almost continuous muscle cramps and painless muscle contractions eight years after being diagnosed as having rheumatoid arthritis. Both cramps and contractions were present at rest, were accentuated by stress and disappeared during sleep. By night-fall the patient was plunged into an extremely disabling condition due to the continuous cramps present in the orofacial, trunk, neck and limb musculature. He even had difficulty speaking. EMG studies demonstrated that both cramps and painless contractions appeared synchronously in muscles innervated by different peripheral nerves. A state of central hyperexcitability is the probable cause of this clinical picture which has remained unchanged over the last six years.

Aged

Involvement of the interleukin 2 pathway in the rearrangement and expression of both alpha/beta and gamma/delta T cell receptor genes in human T cell precursors.

In this report, we have undertaken the phenotypic, functional and molecular characterization of a minor (less than 5%) subpopulation of adult thymocytes regarded as the earliest intrathymic T-cell precursors. Pro-T cells were immunoselected and shown to express different hematopoietic cell markers (CD45, CD38, CD7, CD5) and some activation-related molecules (4F2, Tr, HLA class II), but lack conventional T cell antigens (CD2-1-3-4-8-). TCR-gamma RNA messages are already expressed at this early ontogenic stage, while alpha and beta chain TCR genes remain in germline configuration. In vitro analyses of the growth requirements of pro-T cells demonstrated the involvement of the IL-2 pathway in promoting their proliferation and differentiation into CD3+ CD4+ or CD8+ mature thymocytes. Moreover, during the IL-2-mediated maturation process rearrangements and expression of both alpha and beta chain TCR genes occurred, and resulted in the acquisition of alpha/beta as well as gamma/delta (either disulphide-linked or non-disulphide-linked) heterodimeric TCR among the pro-T cell progeny.

Antigens, Differentiation, T-Lymphocyte

Alpha/beta heterodimeric T-cell receptor expression early in thymocyte differentiation.

The differentiation of T lymphocytes inside the thymus results in the acquisition of MHC-restricted specific functions mediated by clonally distributed alpha/beta heterodimeric T-cell receptors (TcR). Genes encoding the alpha and beta subunits of the clonotypic receptor (Ti) are rearranged during thymic ontogeny and expressed in association with the monomorphic CD3 complex. The regulation of the expression of functional TcR along T-cell development is thus crucial to establish the ontogenic events involved in the acquisition and selection of T-cell repertoires. Current views support that CD3-alpha/beta heterodimers are acquired late in ontogeny on developing thymocytes already expressing CD4 and/or CD8 surface molecules, whereas CD4- CD8- early precursors, representing the major population in the embryonic thymus, do not yet express the alpha/beta TcR. However, a novel CD3-associated gamma/delta heterodimer has been recently identified on the surface of this "double negative" subset both in thymocytes and in MHC-unrestricted peripheral T cells, suggesting that alpha/beta and gamma/delta heterodimeric receptors are independently expressed on the surface of distinct thymic subpopulations during T-cell development. In contrast to these results, we report here that a major proportion of CD3+1-4-8- adult human thymocytes, included within the early "double negative" subset, express alpha/beta heterodimeric receptors, as assessed by flow cytometric analysis using a frame-work monoclonal antibody (WT.31) against the alpha/beta TcR complex. These and previous data showing that CD3+1-4-8- "double negative" thymocytes constitute a functional intermediate ontogenic stage in the differentiation of CD3+1-4+8-/CD3+1-4-8+ mature T cells from CD3-1-4-8- early prothymocytes further support the relevance of the CD3+1-4-8- transitional subset as immediate intrathymic precursors of alpha/beta TcR-bearing mature T cells. Therefore, developmental regulation of alpha/beta TcR expression was analyzed at the DNA, RNA, and protein levels in those different thymic subpopulations, defined by both functional and phenotypic criteria. Our results demonstrate that multiple Ti beta gene rearrangements and beta RNA messages are already evident at the early prothymocyte stage. Moreover, expression of relative levels of both Ti alpha and Ti beta functional RNA transcripts, similar to those observed in mature thymic cells, were also present in CD3+1-4-8- thymocytes. According with these data, immunoprecipitation analysis using a specific anti-Ti alpha antisera revealed that both alpha and beta molecules are expressed on CD3+ "double negative" and mature thymocytes, but not in prothymocytes

Antigens, Differentiation, T-Lymphocyte

Selective expansion of a CD3+CD4-CD8- subpopulation in clinical groups associated with human immunodeficiency virus infection.

T lymphocytes (CD3+) without expression of CD4/CD8 surface antigens have recently been described in the thymus and peripheral lymphoid organs. We have conducted a retrospective analysis of the literature, seeking quantitative variations in this T-cell subset in normal heterosexual controls, and in risk, pre-AIDS, and AIDS groups, by means of the subtraction [CD3-(CD4+CD8]) and the ratio 100 X [CD3-(CD4+CD8])/CD3. Dramatic T lymphocytopaenia in AIDS patients and the progressive decay of CD4+ lymphocytes and increase of CD8+ lymphocytes throughout the clinical spectrum of HIV infection have been confirmed. Furthermore, we hereby demonstrate the selective expansion of CD3+CD4-CD8- lymphocytes, directly related to the clinical state in different clinical groups of infected people when compared with controls (P less than 0.05). The inverse relationship between the CD3+CD4-CD8- cell subset and other mature T-cell subsets, mainly CD4+ (r = -0.49; P less than 0.01), suggests the existence of mutual regulatory interactions. These in vivo results, which are in agreement with those obtained in long-term infected cultures, cannot be explained by direct cytopathic effects of the virus on the very few infected cells. Thus, the implication of the expansion of these functional precursors on the prognosis for infected people, and the paradoxes of the immunodeficiency, such as lymphoproliferation and autoimmune features, are discussed.

AIDS-Related Complex

[Some Spanish contributions to anesthesiology at the end of the XIXth century].

At the end of the XIXth century some original, although poorly known, Spanish contributions were made on the field of inhalational general anesthesia that are important for the history of our specialty in Spain. The purpose of this work is to report the Spanish contributions to inhalational general anesthesia made during the chronological period that coincides with the locus-regional anesthetic techniques grown after the appearance of cocaine in 1884.

Anesthesiology