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Biomedical subjects

J Dominguez

Publications and source records attributed to J Dominguez.

At least 73 records · Page 4Linked to original sources

Analysis of monoclonal antibodies reactive with the porcine CD4 antigen.

As result of the First International Swine CD Workshop, four monoclonal antibodies (mAb) (#002, 054, 118, and 127) clustered closely to the internal standard anti-porcine CD4 mAb, 74-12-4. To further characterize the relationship between these mAb, they were used in flow cytometry to inhibit binding of 74-12-4 to porcine lymphocytes. mAb #002 (74-12-4) and #054 (PT90) completely inhibited, while mAb #127 (10.2H2) and #118 (b38c6) partially inhibited (57% and 77% respectively), the binding of 74-12-4. Furthermore, none of the mAbs bound to a 74-12-4 negative strain of pigs. We conclude that the four mAb bind to the same, or a closely related, epitope on porcine lymphocytes.

Animals↗

Analyses of monoclonal antibodies reactive with porcine CD5.

Among all monoclonal antibodies (mAbs) analyzed in the first porcine CD workshop, six mAbs showed reactivity to the porcine CD5 antigen (workshop mAbs 067, 068, 069, 070, 071 and 119). Because of lack of immunoprecipitation studies for five (067, 068, 069, 070 and 071) out of the six mAbs, only one mAb (119) could be definitely characterized as mAb against the monomeric 63 kDa porcine CD5 antigen. The five other mAbs included in this cluster are characterized by an identical labelling pattern in FCM and competition of the CD5 epitope recognized by mAb 119. These mAbs were allocated to the wCD5 subcluster.

Animals↗

Analyses of monoclonal antibodies reactive with porcine CD6.

Amongst the monoclonal antibodies (mAbs) submitted to the first porcine CD workshop, two mAbs (workshop numbers 055 and 120) could be identified to recognize the porcine CD6 analogue. Both mAbs seemed to be highly T-cell specific and showed neither reactivity with cells of the myeloic lineage nor with B lymphocytes. The observed molecular mass of the antigen precipitated by mAb 120 of 110 kDa confirmed this classification. Without molecular analyses of the antigen recognized by mAb 055, but similar staining pattern in FCM compared with 120, mAb 055 was allocated to the wCD6 subcluster.

Animals↗

Analyses of mAb reactive with porcine CD8.

Among all mAb submitted to the first porcine CD workshop, based on FCM analyses six mAb could be identified to recognize the porcine CD8 analogue (workshop Nos. 004, 051, 052, 053, 108 and 109). In immunoprecipitation studies three mAb (Nos. 004, 108 and 109) recognized an antigen with an apparent molecular mass of about 35 kDa under reducing conditions and about 70 kDa under non-reducing conditions. The molecular masses of the antigens recognized by the three other mAb (Nos. 051, 052 and 053) are still unknown. Epitope analyses performed by blocking experiments led to the determination of two CD8 epitopes: CD8a and CD8b. CD8a is recognized by mAb Nos. 004, 051 and 052, and CD8b by Nos. 053, 108 and 109.

Animals↗

Analysis of mAb reactive with the porcine SWC1.

Among 54 mAb determined to be reactive with porcine T lymphocytes and/or activation antigens, eight mAb (workshop Nos. 005, 031, 080, 091, 092, 093, 094 and 110) derived from different laboratories grouped together in the T11 cluster and were ordered into the SWC1. One mAb (No. 111) which belong also to this group was lost during the workshop. The SWC1 antigen is a molecule expressed on the majority of leukocytes, resting T lymphocytes, monocytes and granulocytes, but not on B lymphocytes. On T lymphocytes it is down-regulated after activation. The molecular mass of the antigen is unknown. Epitope analyses revealed that seven out of the nine mAb recognized similar epitopes on the SWC1 molecule.

Animals↗

Changes in systemic blood pressure and cardiac rhythm induced by therapeutic compression of the trigeminal ganglion.

Percutaneous compression of the trigeminal ganglion, which is currently being used for the control of trigeminal neuralgia, induces marked intraoperative elevations of the systemic blood pressure and heart rate changes, which may increase the risk of cardiovascular complications. We have analyzed the characteristics of the arterial hypertensive response and the cardiac rhythm changes induced by percutaneous compression of the trigeminal ganglion in 42 consecutive, unselected patients undergoing operations for essential trigeminal neuralgia under three different regimens of anesthesia. The first 22 patients (Group 1) underwent operations under brief general anesthesia without endotracheal intubation. The following 10 patients (Group 2) had general anesthesia with intubation and mechanical ventilation and received larger doses of hypnotic and analgesic agents. Finally, 10 more patients (Group 3), who had general anesthesia with intubation, underwent local anesthetic blockade of Meckel's cave (injection of 1 ml of 1% lidocaine) before ganglion compression. Foramen ovale puncture elicited bradycardia in the majority of the patients of Groups 2 and 3, but only four patients (18%) of Group 1 showed bradycardia. Ganglion compression caused marked tachycardia in all patients of Groups 1 and 2; about one-third of the patients also had extrasystoles. By contrast, patients of Group 3, who had local anesthetic blockade of Meckel's cave before ganglion compression, did not develop tachycardia or extrasystoles. Foramen ovale puncture elicited marked elevations of the systemic blood pressure in all patients. Ganglion compression further increased blood pressure, except in patients of Group 3, who had local anesthetic blockade of Meckel's cave.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

[Laparoscopic gastrostomy].

Feeding gastrostomy is a method for treating patients unable to ingest food orally. After our experience and good results with the Janeway technique, we decided to perform it by the laparoscopic method. Our first excellent results in seven patients suggest that laparoscopic Janeway gastrostomy is an easy, rapid and efficient surgical technique.

Aged↗

Intestinal leiomyomas: angiographic imaging.

A retrospective angiographic study was done on 19 patients with solitary leiomyomas of the small bowel, one patient with multiple leiomyomas, and two patients with leiomyosarcomas. All patients had gastrointestinal bleeding but no small bowel obstruction. Selective mesenteric angiography was the main diagnostic procedure. Tumoral hypervascularity and dense capillary staining within a well-circumscribed mass were present in all patients. Prominent and irregular feeding arteries were seen in 19 cases and prominent draining veins in 20. Contrast extravasation indicating the site of active bleeding was shown in two patients.

Adult↗

Localization of African swine fever viral antigen, swine IgM, IgG and C1q in lung and liver tissues of experimentally infected pigs.

An immunohistological study was carried out on lungs and livers of pigs experimentally infected with two different African swine fever virus (ASFV) isolates. ASFV antigen, swine immunoglobulins (IgM and IgG) and (Clq) complement were demonstrated in both organs at different stages of infection. The ASFV antigen was mainly found in mononuclear phagocytic system (MPS) cells. Immunoglobulins and complement were observed in plasma, infected and non-infected phagocytic cells and cell debris. These findings suggest the presence, in acute infection, of immune complexes which may be involved in immunopathogenic mechanisms.

African Swine Fever↗

Detection of African horsesickness virus in infected spleens by a sandwich ELISA using two monoclonal antibodies specific for VP7.

A sandwich enzyme-linked immunsorbent assay (ELISA) for rapid detection of African horsesickness virus (AHSV) in infected spleens or cell culture supernatant was developed. This method uses two monoclonal antibodies (MAbs) which recognize two non-overlapping epitopes of the major core protein (VP7) to coat the solid phase, and one labeled with biotin as second antibody. This ELISA was evaluated for its ability to detect AHSV in infected spleens resulting in a sensitivity of 97.4% and a specificity of 100% compared with virus isolation in cell culture, and can be used for the detection of the nine different AHSV serotypes.

African Horse Sickness↗

Role of protein kinase C in the regulation of prostaglandin synthesis in human endothelium.

The present study specifically addresses the role of protein kinase C (PKC) activation in human endothelial cell Ca2+ mobilization, a response that is functionally coupled to the production of the potent arachidonate (AA) metabolite, prostacyclin (PGI2). Phorbol 12-myristate 13-acetate (PMA), alpha-thrombin, and sodium fluoride (NaF), a direct G-protein activator, produced a rapid and time-dependent translocation of PKC from the cytosol to the membrane. Activation of PKC by brief pretreatment of human umbilical vein endothelial cell (HUVEC) monolayers with PMA resulted in the inhibition of NaF-induced inositol phosphate increases and attenuation of both alpha-thrombin- and NaF-activated increases in intracellular Ca2+ (Ca2+i). Ca2+ mobilization induced by ionophore A23187 was not affected by PKC preactivation, suggesting PKC-dependent negative feedback inhibition of phosphatidylinositol (PI)-specific phospholipase C (PLC). Agonist-stimulated AA release and PGI2 synthesis in PMA-pretreated cultured human endothelial cells, however, was potentiated, and the enhanced PGI2 synthesis produced by A23187, NaF, and alpha-thrombin was dependent upon the dose of PMA. Treatment of HUVEC monolayers with an intracellular Ca2+ chelator, 1,2-bis(2-aminophenoxy)ethane-N,N,N'N'-tetraacetic acid-acetoxymethylester (BAPTA-AM), dramatically reduced alpha-thrombin-, NaF-, and A23187-induced PGI2 synthesis, demonstrating the importance of Ca2+i availability in PGI2 synthesis. BAPTA pretreatment did not inhibit PMA-induced PKC activation, and BAPTA-mediated inhibition of agonist-stimulated PGI2 synthesis was partially attenuated by prior PMA pretreatment. Staurosporine, a potent PKC inhibitor, at concentrations that inhibited PKC-induced phosphorylation of histone-1, augmented both alpha-thrombin- and NaF-induced production of inositol phosphates but markedly inhibited alpha-thrombin-, NaF-, and A23187-induced PGI2 synthesis. The downregulation of PKC activity by prolonged PMA treatment (18 h) produced similar inhibition of PGI2 synthesis by these agonists (approximately 50% inhibition). These studies indicate that the integrated phospholipase A2 and PLC activities are under complex regulation by factors that include both PKC activation and [Ca2+i]. PKC exerts dual effects on prostaglandin synthesis via negative regulation of Gp-coupled PI-specific PLC and positive feedback regulation of AA release and PGI2 synthesis. PKC is thus a critical determinant in the regulation of human endothelial cell prostaglandin synthesis by both receptor-mediated and G-protein-dependent cellular activation.

Aequorin↗

Gp-regulated phosphoinositide hydrolysis in turkey and human erythrocytes exposed to fluoride ion: relationship to calcium influx.

Previous studies have demonstrated that although both mammalian and avian erythrocytes express an inducible inositol bisphosphate-specific phospholipase C, only the latter possess the guanine nucleotide-binding protein (Gp) that regulates this activity. In confirmation of previous reports, turkey erythrocyte plasma membranes responded to guanosine 5'-0-(3-thio)triphosphate (GTP-gamma-S) and fluoroaluminates with hydrolysis of phosphoinositides, release of inositol phosphates, and generation of diacylglycerol, whereas human erythrocyte plasma membranes exhibited no such changes when incubated with known activators of guanine nucleotide regulatory proteins. We next contrasted responses of intact turkey and human erythrocytes to fluoroaluminates to develop a model to investigate the cellular effects of Gp activation. When turkey erythrocytes were exposed to fluoroaluminates, cellular levels of diacylglycerol and phosphatidic acid rapidly increased as phosphoinositides were hydrolyzed. The alterations in the lipid composition of turkey erythrocytes effected by fluoroaluminates were remarkable; phosphatidic acid levels increased over 30-fold, whereas levels of polyphosphoinositides were decreased to less than 10% of those present before stimulation. In contrast, fluoroaluminates caused only minor alterations in the diacylglycerol and phospholipid content of intact human erythrocytes. To define the role of inositol-specific phospholipase C activation in the transmembrane conveyance of extracellular Ca++, we compared the influx of extracellular Ca++ in human and turkey erythrocytes exposed to fluoroaluminates. Fluoroaluminates initiated a sustained influx of extracellular 45Ca++ into turkey, but not human, erythrocytes. These results provide strong support for the hypothesis that Gp activation results in an influx of calcium into stimulated cells. Moreover, the data demonstrate that comparison of responses of human and turkey erythrocytes to fluoroaluminates provides a well-defined method for investigating the mechanisms and consequences of Gp activation in intact cells.

Animals↗

Clinical and ultrastructural analysis of failures in chemonucleolysis.

A series of 12 patients with failed chemonucleolysis were analyzed to determine the various causes of such failures. The ultrastructural findings observed in laminectomy specimens did not account for the failure in the enzyme treatment. The findings suggested deficient cellular nutrition secondary to enzymatic histolysis of the nucleus pulposus.

Chymopapain↗

MAP-1 and MAP-2 binding sites at the C-terminus of beta-tubulin. Studies with synthetic tubulin peptides.

The interaction of microtubule-associated proteins MAP-1 and MAP-2 with different peptides containing sequences covering the C-terminal region of beta-tubulin isoforms has been analyzed. Our results indicate that MAP-1 and MAP-2 bind to a common sequence within the variable C-terminal region of the different beta-tubulin isoforms, while MAP-2 also interacts with the subdomain beta (422-434) of the constant region, in agreement with previous results (Maccioni, R.B., Rivas, C., & Vera, J.C. (1988) EMBO J. 7, 1957-1963). The productive interaction of MAP-2 with the latter domain appears to be involved in the assembly of microtubules.

Amino Acid Sequence↗

Inhibitory effects of phospholipase D on chondrogenesis in vitro.

Mesenchymal cells from the wing buds of stage 24 chick embryos undergo differentiation to cartilage when plated at high density. Treatment of these cultures with phospholipase D resulted in inhibition of chondrogenesis. Phospholipase D treatment (which produces phosphatidic acid from membrane phospholipids) was found to affect cell proliferation and to dramatically increase intracellular free calcium levels and inositol phosphate production. Intracellular free Ca2+, mobilized as a result of phosphatidylinositol phosphate hydrolysis, may therefore inhibit chondrogenesis in embryonic mesenchymal cells.

Animals↗

Sodium fluoride induces phosphoinositide hydrolysis, Ca2+ mobilization, and prostacyclin synthesis in cultured human endothelium: further evidence for regulation by a pertussis toxin-insensitive guanine nucleotide-binding protein.

The role of guanine nucleotide-binding proteins in the induction of prostacyclin synthesis by stimulated endothelial cells is incompletely understood. We report that sodium fluoride (NaF), a potent activator of cellular guanine nucleotide-binding proteins, affected time- and concentration-dependent generation of prostacyclin (PGI2) by cultured human umbilical vein endothelial cells without evidence of cellular toxicity detected by 51Cr or lactate dehydrogenase release. PGI2 synthesis by NaF-stimulated endothelial cells was associated with increases in arachidonate release, phosphoinositide hydrolysis, generation of inositol phosphates, and accumulation of diacylglycerol. These responses to NaF, as well as alpha-thrombin-mediated responses, were not dependent upon the availability of extracellular free Ca2+ but were associated with the mobilization of stored intracellular Ca2+ detected by the luminescence of the photoprotein aequorin. Neither PGI2 synthesis nor Ca2+ responses following alpha-thrombin or NaF stimulation were inhibited by pretreatment of cells with the islet activating protein from Bordetella pertussis but were significantly attenuated by the G protein inhibitor GDP beta S in permeabilized cells. Our results are compatible with a model wherein NaF directly activates a phosphoinositidase-linked guanine nucleotide regulatory protein, Gp, in human umbilical vein endothelial cell monolayers. This activation results in phosphoinositide hydrolysis, Ca2+ mobilization, arachidonate release, and subsequent functional activation, assessed by PGI2 release. Biologically relevant agonists such as alpha-thrombin may exert their influence on arachidonate metabolism, in part, by promoting receptor-dependent activation of this G protein.

Adenosine Diphosphate Ribose↗

Unique amino acid substitutions in the capsid proteins of foot-and-mouth disease virus from a persistent infection in cell culture.

Maintenance of a persistent foot-and-mouth disease virus (FMDV) infection in BHK-21 cells involves a coevolution of cells and virus (J. C. de la Torre, E. Martínez-Salas, J. Díez, A. Villaverde, F. Gebauer, E. Rocha, M. Dávila, and E. Domingo, J. Virol. 62:2050-2058, 1988). The resident FMDV undergoes a number of phenotypic changes, including a gradual decrease in virion stability. Here we report the nucleotide sequence of the P1 genomic segment of the virus rescued after 100 passages of the carrier cells (R100). Only 5 of 15 mutations in P1 of R100 were silent. Nine amino acid substitutions were fixed on the viral capsid during persistence, and three of the variant amino acids are not represented in the corresponding position of any picornavirus sequenced to date. Cysteine at position 7 of VP3, that provides disulfide bridges at the FMDV fivefold axis, was substituted by valine, as determined by RNA, cDNA, and protein sequencing. The modified virus shows high buoyant density in cesium chloride and depicts the same sensitivity to photoinactivation by intercalating dyes as the parental FMDV C-S8c1. Amino acid substitutions fixed in VP1 resulted in altered antigenicity, as revealed by reactivity with monoclonal antibodies. In addition to defining at the molecular level the alterations the FMDV capsid underwent during persistence, the results show that positions which are highly invariant in an RNA genome may change when viral replication occurs in a modified environment.

Amino Acid Sequence↗