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Biomedical subjects

M Rojas

Publications and source records attributed to M Rojas.

At least 73 records · Page 4Linked to original sources
PubMed1998

Increase in asialoganglioside- and monosialoganglioside-reactive antibodies in chronic Chagas' disease patients.

Antibodies reactive with the core glycan of asialoganglioside (GA1), monosialoganglioside (GM1), and disialoganglioside (GD1a) were studied in human sera. In healthy individuals, GA1-, GM1-, and GD1a-reactive antibodies were mainly of the IgM class, but also of the IgA and IgG classes, and were present at low titers in the serum of 68%, 79%, and 91% of the individuals studied, respectively. Levels of anti-GA1 and anti-GM1 antibodies, mainly of the IgA and IgG classes, were significantly elevated (P < 0.001) in 62% and 72% of subjects, respectively, chronically infected with Trypanosoma cruzi, with no association found with the degree of myocardial damage. No significant increase in anti-GA1 and anti-GM1 antibodies was found in dilated cardiomyopathy patients. The level of anti-GD1a antibody was not significantly different between healthy controls and chronic chagasic or dilatatory cardiomyopathy patients. Since the peripheral nervous system is very rich in gangliosides, it is possible that the increases in GA1- and GM1-specific antibodies that develop during chronic T. cruzi infection are involved in the pathology of peripheral neuropathy in Chagas' disease.

Animals↗
PubMed1998

High benzo[a]pyrene diol-epoxide DNA adduct levels in lung and blood cells from individuals with combined CYP1A1 MspI/Msp-GSTM1*0/*0 genotypes.

Levels of anti-benzo[a]pyrene diol-epoxide DNA adducts were analysed by high-pressure liquid chromatography/fluorimetric detection in non-tumorous lung tissues from 20 lung cancer patients and in white blood cells from 20 polycyclic aromatic hydrocarbon exposed coke oven workers. All were current tobacco smokers. CYP1A1 mutations (MspI at 6235 nt, Ile-Val462) and GSTM1 deletion polymorphisms in each individual were analysed in genomic DNA by PCR/restriction fragment length polymorphism. Independently of the CYP1A1 genotype (1) all 23 samples in the two groups with non-detectable adducts (< 0.2 per 10(8) nt) were of GSTM1 active genotype; (2) the 17 samples with detectable adducts (> or = 0.2 per 10(8) nt) in the two groups were GSTM1*0/*0. The difference in adduct levels between GSTM1*0/*0 and GSTM1 active genotype was highly significant (p < 0.00005). Among GSTM1-deficient individuals (n = 17), a subgroup of 14 individuals with CYP1A1*1/*1 (wild-type, n = 7) or heterozygous genotype (*1/*2A or *1/*2B, n = 7) showed low levels of BPDE DNA-adducts (range: 0.2-1.3 per 10(8) nt). (3) Three individuals with the rare combination CYP1A1*2A/*2A or *2A/*B and GSTM1*0/*0 showed significantly higher adduct levels (median: 17.4 adducts/10(8) nt, range 1.9-44; p = 0.017). Therefore, combination of homozygous mutated CYP1A1 and GSTM1*0/*0 genotypes lead, at a similar or even lower smoking dose, to a stronger increase of anti-benzo[a]pyrene diol-epoxide DNA adduct levels than found in individuals with CYP1A1 and GSTM1 wild-type. These data provide a mechanistic understanding of epidemiological studies that correlated these 'at risk' genotypes with increased smoking-related lung cancers.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗
PubMed1998

[Essential fatty acid content of maternal erythrocyte phospholipids. A study in preterm and full-term human newborns].

BACKGROUND: A high concentration of arachidonic acid in maternal erythrocytes and trophoblast could have a role in pre term deliveries. AIM: To study the fatty acid composition of long chain fatty acids from erythrocytes of mothers who gave birth to pre term and full term infants. PATIENTS AND METHODS: Thirty three healthy women that gave birth to healthy newborns in a public hospital were studied. Twenty two had pre term (34 weeks) and 11 full term (40 weeks) deliveries. The fatty acid profile of phospholipids isolated from erythrocytes of these women was analyzed by gas-liquid chromatography. RESULTS: Compared to women giving birth to full term infants, phospholipids of women giving birth to pre term infants had a higher content of arachidonic acid (20:4 omega 6) and all the species of omega 6 fatty acids. They also had a lower concentration of palmitic and eicosapentanoic (20: 5 omega 3) acids and thus a higher arachidonic acid/eicosapentanoic acid ratio. CONCLUSIONS: A high arachidonic acid content in phospholipids of erythrocytes could be a risk factor or predictive marker for pre term deliveries.

Adult↗
PubMed1998

[Multi-plane transesophageal echocardiography in cardiac angiosarcoma. Apropos of a case].

The authors report the case of a right atrial angiosarcoma extending to the right ventricle, presenting with a massive pericardial effusion, in a 30 year old man. The patient underwent surgical ablation of the tumour with right atrial reconstruction, tricuspid valvuloplasty, plicature of the right ventricle and a right coronary artery bypass graft with complementary chemotherapy. This case illustrates the decisive contribution of multiplane transoesophageal echocardiography in the preoperative investigation, providing accurate information about the site and extension of the tumour, and even about the malignant character of the pathology. Magnetic resonance imaging, though useful, did not provide any further information in this case. Unfortunately, thoracic CT scan, performed 15 days after surgery, showed the presence of pulmonary metastases, confirming the very poor prognosis of these tumours.

Adult↗
PubMed1997

The preferential induction of a Th1 immune response by DNA-based immunization is mediated by the immunostimulatory effect of plasmid DNA.

In the present study, we have investigated the T cell response to the HBsAg, normally secreted as multivalent particles, and to beta-galactosidase, a cytoplasmic antigen, delivered as plasmid DNAs. We found that cytokines characteristic of a Th1 phenotype are produced in mice immunized by these plasmid DNAs. Using repeated injections of low doses of purified antigen, we demonstrated that neither prolonged presence of the antigen nor site of immunization resulted in an immune response with characteristics resembling those obtained with DNA-mediated immunization. Analysis of immune responses induced in mice by coinjection of plasmid DNA and beta-galactosidase or HBsAg demonstrated that the coinjected DNA stimulated a Th1 response against the injected antigen. These data therefore strongly suggest that the strong immune response obtained after intramuscular DNA immunization was due to the adjuvant effect of the plasmid DNA which is also responsible for the selective activation of CD4(+) T cells with a Th1 phenotype.

Adjuvants, Immunologic↗
PubMed1997

Differential induction of apoptosis by virulent Mycobacterium tuberculosis in resistant and susceptible murine macrophages: role of nitric oxide and mycobacterial products.

Resistance and susceptibility of macrophages to mycobacteria are under the control of the Bcg/Nramp1 gene, which also controls the NO- production in response to macrophage activators. There is recent evidence indicating that mycobacteria induces apoptosis in infected macrophages. Using murine macrophage lines, congenic at the Bcg/Nramp1 gene, this report shows that B10R are more prone than B10S macrophages to undergo apoptosis after exposure to live virulent Mycobacterium tuberculosis H37Rv (Mtb) or PPD, as determined by cell viability, DNA fragmentation, hypoploidy, and the terminal deoxynucleotide transferase dUTP-biotin nick-end labeling assay. Induction of apoptosis correlated with NO- production. Aminoguanidine and anti-TNF-alpha inhibited NO- production and apoptosis. B10R and B10S macrophages were equally affected by sodium nitroprusside, a donor of NO-, but its effect, mainly in B10R cells, was enhanced by the presence of Mtb. Nonvirulent mycobacteria induced lower levels of NO- and did not cause cell death. Killed Mtb, mannose-capped lipoarabinomannan (ManLAM), and LPS rescued macrophages from apoptosis albeit induce NO-. These findings suggest the existence of opposite pathways: metabolically active mycobacteria promotes apoptosis whereas their structural components inhibit it. Apoptosis may be a critical mechanism by which Nramp1 gene controls the macrophage infection with virulent mycobacteria.

Animals↗
PubMed1997

An alternative to phosphotyrosine-containing motifs for binding to an SH2 domain.

Shc is an important signalling protein whose overexpression leads to cell transformation in NIH 3T3 fibroblasts. Although the formation of Shc/Grb2 complexes involving Shc tyrosine residue 317 is necessary to induce this transformation, the Shc proteins in these Shc-overexpressing cells are not substantially tyrosine-phosphorylated. This observation led to our hypothesis that the non-phosphorylated Tyr317-containing region of Shc might have specific affinity for the Grb2 protein. We show here that cell-permeable peptides encompassing the Shc Tyr317 region, 312FDD-PSYVNVQNL323, can bind to the SH2 domain of Grb2 regardless of the state of tyrosine phosphorylation. When delivered into cells, both phosphorylated and non-phosphorylated Shc peptides inhibit growth factor-induced Shc/Grb2 protein-protein interaction. The non-phosphorylated Shc peptides with single point mutations at Asp313, Asp314, or Tyr317 are inactive, suggesting that these residues play an important role in Grb2 protein recognition. Our findings represent the first paradigm of the specific interaction between an unphosphorylated tyrosine-containing region and an SH2 domain and have important implications for understanding the mechanism of cell transformation by Shc overexpression.

3T3 Cells↗
PubMed1997

Antibody levels against alpha-galactosyl epitopes in sera of patients with squamous intraepithelial lesions and early invasive cervical carcinoma.

We measured serum levels of anti-Gal(alpha 1-->3)Gal and anti-Gal(alpha 1-->2)Gal antibodies in 89 and 91 women, respectively, by using ELISA. These patients had cervical intraepithelial neoplasia (CIN) grades 1 to 3 and early invasive cervical carcinoma (ICC). Our objective was to compare anti-alpha-galactosyl antibody levels among them and with those of normal controls. High levels of anti-Gal(alpha 1-->2)Gal antibodies were detected in 22% of patients (P = 0.006). The mean level was 1.6 times greater than that of controls, without difference among subgroups. Thirty percent of patients had abnormally high anti-Gal levels (P = 0.001). Mean levels were twofold greater than the mean control value. Subsets with human papillomavirus/CIN 1 and CIN 2-3 had high immunoreactivity (P = 0.004). Both antibodies showed a significant correlation (r = 0.53, P < 0.00001). We conclude that 22 to 30% of patients with CIN 1-3 showed significantly high levels of anti-alpha-galactosyl antibodies. This seroreactivity might be related to the abnormal expression of alpha-galactosyl residues at some point of the natural history of human papillomavirus infection of the uterine cervix, suggesting an active immune response by natural antibodies against this virus. Further studies are needed to determine whether anti-alpha-galactosyl antibodies confer protection in human papillomavirus infection.

Adult↗
PubMed1997

Depressed autoantibody synthesis in Trypanosoma cruzi-infected rats born to mothers undergoing this infection during pregnancy.

Earlier work indicated that Trypanosoma cruzi infection in pregnant rats decreased the amount of myocardial damage that developed in their chronically infected offspring. Given the suspected role of autoimmune mechanisms in the generation of chronic myocarditis, we evaluated whether this maternal intervention was likely to affect the synthesis of autoantibodies in infected young. Autoantibodies were investigated against molecules exhibiting cross-reactivity with T. cruzi antigens or not, that is cerebroside sulphate (sulphatide) and actin, respectively. Female '1' rats (75 days old) that had been mated with syngeneic sires were separated into two groups, one challenged with living trypomastigotes at 7, 14 and 21 days following mating, and the other one given physiologic saline at the same intervals. At the time of weaning, offspring were injected with 10(6)/T. cruzi to constitute two infected groups: young born to infected mothers (InMoTc) and young delivered by uninfected mothers (CoMoTc). Serum antibodies were investigated by ELISA at 30 and 60 days post-infection, which represents acute and chronic infection, respectively. T. cruzi infection was associated with the production of anti-sulphatide antibodies, but the phenomenon was significantly less evident in InMoTc young and virtually unnoticeable during their chronic infection. Unlike the anti-sulphatide results, levels of anti-actin antibodies showed no differences between CoMoTc and InMoTc rats when compared during acute or chronic infection. The decreased production of anti-sulphatide autoantibodies of InMoTc offspring may be due to a modification of the immune repertoire of offspring because of the contact with parasite antigens during ontogeny.

Actins↗
PubMed1997

Inhibition of virulent Mycobacterium tuberculosis by Bcg(r) and Bcg(s) macrophages correlates with nitric oxide production.

The Nramp1 gene controls macrophage resistance or susceptibility to several intracellular microorganisms; however, there is conflicting evidence regarding its role during infection with virulent Mycobacterium tuberculosis. Nitric oxide (NO) is a potent antimycobacterial agent produced by macrophages, which is also regulated by Nramp1. The in vitro ability of B10R (resistant) and B10S (susceptible) murine macrophages to inhibit M. tuberculosis H37Rv and to produce NO in response to infection and interferon-gamma (IFN-gamma) was compared. Infected B10R macrophages inhibited [3H]uracil incorporation by M. tuberculosis and produced higher amounts of NO than did B10S macrophages. IFN-gamma increased the inhibitory activity of both cells. Inhibition of M. tuberculosis by IFN-gamma-activated B10R macrophages was reversed by N(G)-monomethyl-L-arginine (N(G)MMA). L-arginine restored NO production and increased the antimycobacterial activity by IFN-gamma-stimulated N(G)MMA-treated macrophages. The Bcg/Nramp1 gene may regulate macrophage resistance or susceptibility to virulent M. tuberculosis by a differential capability of these cells to produce NO.

Animals↗
PubMed1997

Immunomagnetic selection of purified monocyte and lymphocyte populations from peripheral blood mononuclear cells following cryopreservation.

Cryopreservation is a method commonly used to store human blood samples. We sought to determine if cryopreserved peripheral blood mononuclear cells (PBMC) could be separated effectively into distinct populations by using monoclonal antibodies and immunomagnetic microspheres. PBMC obtained from healthy blood donors and from human immunodeficiency virus-infected subjects were cryopreserved for as long as 18 months. Recovered cells were separated into CD14+ monocytes and CD4+ T-cell subsets by immunomagnetic selection. Flow cytometry analysis indicated >95% depletion of monocytes from PBMC following immunomagnetic selection with anti-CD14. A highly enriched population of CD4+ T cells was obtained from the CD14-depleted cell fraction by using an anti-CD4 monoclonal antibody and detachable immunomagnetic beads. The CD4+ T cells were subsequently separated into CD4+ CD45RO and CD4+ CD45RA fractions. Each fraction contained >90% enrichment for the respective subpopulation and <5% of the reciprocal subpopulation. No significant differences in cell surface expression of leukocyte markers, in efficiency of selection of PBMC subpopulations, or in mitogen-induced proliferation were detected in freshly isolated or cryopreserved cells. Efficient recovery of cryopreserved specimens means that targeted assays can be performed on selected, prospectively stored samples once clinical endpoints have been achieved.

Antibodies, Monoclonal↗
PubMed1997

[Sensitivity of spiral volumetric TC in the diagnosis of hepatic metastases: advantages of the dual phase technic].

In this report the diagnostic possibilities of volumetric Spiral CT in hepatic metastases detection are discussed. Spiral CT presents numerous advantages when compared to the conventional one; these increase its sensitivity in hepatic metastases diagnosis of approximately 10% (Conventional CT sensitivity 80-85%; Spiral CT sensitivity 90-95%). "Dual phase" technique allows hypervascular metastases diagnosis, often isodense during portal venous phase, hence not detectable with conventional CT. Nonetheless, helical CT for hepatic metastases detection is not yet common, due to high exams costs and more often due to the limited availability of equipment.

Humans↗
PubMed1996

Controlling epidermal growth factor (EGF)-stimulated Ras activation in intact cells by a cell-permeable peptide mimicking phosphorylated EGF receptor.

Epidermal growth factor (EGF)-stimulated Ras activation involves specific interactions between the EGF receptor (EGFR), the adaptor proteins Grb2 and Shc, and the nucleotide exchange factor Sos-1. Study and control of these protein-protein interactions in vivo can be greatly promoted by introducing intracellular reagents that mimic EGFR functions. Here, we showed that a synthetic phosphopeptide encompassing the autophosphorylation site 1068 of EGFR formed a complex with endogenous Grb2 after this peptide was delivered into intact cells by a cell-permeable peptide import technique. Consequently, this intracellular peptide inhibited EGF-induced EGFR/Grb2 associations but not EGFR/Shc or Shc/Grb2 associations. Peptide-mediated disruption of the EGF/Grb2/Sos-1 cascade led to reduced Ras activation and mitogen-activated protein kinase activation. These results indicate that the binding of Grb2 to the phosphorylated Tyr-1068 of EGFR is crucial to the EGF-induced Ras/mitogen-activated protein kinase signaling pathway. The application of cell-permeable peptides to this study demonstrates a useful biochemical tool to probe and control various intracellular processes involved in signal transduction and gene transcription.

3T3 Cells↗
PubMed1996

Intrasubject variability of repeated pulmonary function measurements in preterm ventilated infants.

This study set out to describe the variability and assess the reproducibility of repeated pulmonary function measurements in ventilated preterm infants. We measured tidal volume (VT), lung compliance (CL), and resistance (RL) in 16 infants (mean +/- SD: birthweight 1222 +/- 343 g) during spontaneous breathing and during mechanical ventilation, suppressing breathing efforts by mild hyperventilation. CL and RL were calculated from the equation of motion using linear regression analysis (LR), and by the Mead and Wittenberger method (MW). Flow and transpulmonary pressure were recorded for at least two consecutive periods, after which the esophageal tube was removed and replaced 1 hour later for a second set of recordings. The mean percent change (% delta) between the initial and the repeated measurements with their respective 95% confidence intervals were calculated. Reproducibility was assessed by the intraclass correlation coefficient (ICC) (total agreement = 1, good reproducibility > or = 0.75). The mean % delta between initial and repeat measurements during spontaneous breathing ranged from 11% to 14% for CL and VT, and from 22% to 32% for RL. The variation for RL was even higher when the analysis was done separately for the inspiratory and expiratory phase. CL and VT had good reproducibility (ICC > 0.9), while RL was significantly less reproducible (ICC < 0.75). Measurements obtained from mechanical breaths had less variability than from spontaneous breaths, ranging from 8% to 15% for CL and VT, and from 13% to 21% for RL. Reproducibility assessed by the ICC was good for most measurements during mechanical breaths. The variability and reproducibility of measurements were similar for both methods of analysis during mechanical ventilation, but during spontaneous breathing variability was larger with the MW method than with LR analysis. We concluded that VT and CL were reproducible during spontaneous and mechanical breathing. However, RL measurements were reproducible only during mechanical ventilation. The high variability of RL in spontaneously breathing preterm infants may reduce the clinical usefulness of this measurement for individual patients.

Humans↗
PubMed1996

Cholesterol sulphate-reactive autoantibodies are specifically increased in chronic chagasic human patients.

An antibody reactive with cholesterol sulphate (CS) was characterized in human sera by ELISA, erythrocyte and liposome absorption. This antibody was found evenly distributed between the IgA and IgM classes, and whilst this was present at low titres in the serum of 16% of healthy individuals studied, it was significantly elevated in 78% of Trypanosoma cruzi-infected subjects. No association was found between antibody levels and the degree of myocardial damage. No significant difference in immunoreactivity was found between healthy and chagasic subjects using dehydro-epiandrosterone sulphate and pregnenolone sulphate and cholesterol, ergosterol, lanosterol, stigmastanol, beta-stigmasterol, pregnenolone, prednisolone and dehydroepiandrosterone as antigens, suggesting that in chagasic sera the whole sterol molecule is important for optimal antibody binding. CS-reactive antibodies were easily purified by absorption either with CS-bearing liposomes or with dextran sulphate gel and further elution with 1.5 M NaCl. The optimal pH of CS-antibody interaction was 4.0 with 85% binding at pH 7.0. Polylysine strongly decreased the binding of these antibodies to the corresponding antigen. Furthermore, these antibodies were strongly absorbed by rabbit and guinea pig erythrocyte but not by rat or human erythrocyte. In contrast with anti-sulphatide antibodies, no significant increase in CS-reactive antibodies was found in dilated cardiomyopathies. Whilst CS itself was not detected in T. cruzi lipid extracts, there is an unidentified sulphated sterol, which migrates close to standard CS and which strongly binds chagasic but not control sera. This latter sterol might be acting in chagasic patients as a powerful antigen, triggering specific autoantibody production.

Animals↗