PubMed Health⌕ Search

Biomedical subjects

A M Vázquez

Publications and source records attributed to A M Vázquez.

14 recordsLinked to original sources

RYS1, a foldback transposon, is activated by tissue culture and shows preferential insertion points into the rye genome.

The study of two variable amplicons of rye indicates that RYS1, a mobile element, is activated during tissue culture. We propose that RYS1 could be a foldback (FB) transposon. The FB transposons have been rarely reported in plants; RYS1 is the first described in rye and also the first active plant FB transposon reported. Preferential integration points in the rye genome exist, because the new insertions seem to be located, in all studied cases, in the same genome positions. We assume that RYS1 became active in rye very recently, as different plants from in vivo-growing cultivars showed that these elements were present or absent in the same genomic position in which the in vitro-activated element was found. This high rate of modification in these particular loci, both in the in vivo and in vitro populations, could indicate that probably the mechanisms promoting genetic variability in nature are the same that induce variation in vitro, and the modifications induced by somaclonal variation could be already present in vivo populations.

Culture Techniques↗

Expression of different abscisic acid-responsive genes during somatic embryogenesis in sugarcane (Saccharum officinarum).

We have examined the expression of four genes during somatic embryogenesis in two cultivars of sugarcane, Saccharum officinarum, one drought resistant (JA-605) and the other one sensitive (C-8751), as well as in embryogenic and nonembryogenic tissues treated with abscisic acid (ABA) or after drought stress. Three of the cDNAs probed, from plasmids pMA1049, pMA2005 and CM1, were lea genes and the other one, from a barley hemoglobin gene, was induced in other species under stress conditions. Only transcripts homologous to the pMA2005 and CM1 clones were differentially accumulated during somatic embryogenesis; moreover, the CM1 clone only appeared in somatic embryos of the non-resistant variety. All the lea cDNAs were dramatically increased in the embryogenic tissues treated with ABA, but we observed differences among the accumulation of these mRNAs in the nonembryogenic tissues treated with ABA, and only the pMA2005 transcript appeared. The mRNA homologous to the barley hemoglobin gene appeared in the nonembryogenic tissue and the expression increased after ABA treatment. The desiccation treatments had different effects on the varieties, and the pMA2005 transcripts only appeared in the sensitive variety.

Abscisic Acid↗

[Pilot study with the monoclonal antibody IOR-C5 as a potential agent of radioimmunoscintigraphy in colorectal cancer].

UNLABELLED: IOR C-5 is a G1 immunoglobulin type intact murine monoclonal antibody (MAb) that was developed in the Center of Molecular Immunology in Havana City, Cuba. In immunohistochemical studies, this demonstrated a significant affinity for the epithelial tissues so that it was used in a pilot clinical study to perform a radioimmunoscintigraphy of the colorectal primary tumors and their locoregional recurrences. It was labeled with 99mTc using the Schwarz method, with a > 95% performance. Planar images of the chest, abdomen and pelvis were performed at 10 minutes, 4-6 hours and 18-24 hours post-injection in the anterior and posterior projections and the SPECT was performed 4-6 hours and 18-24 hours post-injection of 1.85 GBq 99mTC. This study has aimed to verify in vivo the capacity of ior-C5 MAb to accumulate in the malignant colorectal lesions. ior-C5 accumulated in 5 out of the 7 patients who were studied and who were suffering from colorectal cancer or in whom there was suspicion of recurrence. There was a negative case of primary tumors, which was an adenocarcinoma in situ in a tubular-papillary adenoma. The second case with a negative radioimmunoscintigraphy was a true negative case. CONCLUSIONS: It can be concluded that even though the number of patients is quite low, ior-C5 fulfilled the expectations of recognizing the epitope expressed in colorectal tumors in an in vivo human environment.

Adenocarcinoma↗

A mouse IgG1 monoclonal antibody specific for N-glycolyl GM3 ganglioside recognized breast and melanoma tumors.

14F7 murine monoclonal antibody (MAb) is an IgG1 immunoglobulin that is generated by immunizing Balb/c mice with GM3(NeuGc) ganglioside hydrophobically conjugated with human very-low-density lipoproteins and in the presence of Freund's adjuvants. 14F7 MAb binds specifically to GM3(NeuGc), whereas neither N-glycolyl or N-acetyl gangliosides, nor a sulfated glycolipid, are recognized as assessed by enzyme-linked immunosorbent assay or immunostaining on thin layer chromatograms. Immunohistochemical studies in fresh tumor tissues showed that 14F7 MAb strongly recognized in antigen expressed in human breast and melanoma tumors.

Animals↗

Syngeneic anti-idiotypic monoclonal antibodies to an anti-NeuGc-containing ganglioside monoclonal antibody.

An IgM monoclonal antibody (MAb), named P3, has the characteristic to react specifically with a broad battery of N-glycolyl containing-gangliosides and with antigens expressed on breast tumors. When this MAb was administered alone in syngeneic mice, an specific IgG anti-idiotypic antibody (Ab2) response was induced, this Ab2 response was increased when P3 MAb was injected coupled to a carrier protein and in the presence of Freund's adjuvant. Spleen cells from these mice were used in somatic-cell hybridization experiments, using the murine myeloma cell line P3-X63-Ag8.653 as fusion partner. Five Ab2 MAbs specific to P3 MAb were selected. These IgG1 Ab2 MAbs were able to block the binding of P3 MAb to GM3(NeuGc) ganglioside and to a human breast carcinoma cell line. Cross-blocking experiments demonstrated that these Ab2 MAbs are recognizing the same or very close sites on the Abl MAb. The five Ab2 MAbs were injected into syngeneic mice and four of them produced strong anti-anti-idiotypic antibody (Ab3) response. While these Ab2 MAbs were unable to generate Ab3 antibodies with the same antigenic specificity than P3 MAb, three of them induced antibodies bearing P3 MAb idiotopes (Ag-Id+ Ab3). These results demonstrated that these Ab2 MAbs are not "internal image" antibodies, but they could define "regulatory idiotopes."

Animals↗

Delineation of the epitope recognized by an antibody specific for N-glycolylneuraminic acid-containing gangliosides.

P3 is a mouse monoclonal antibody (mAb) that binds to several NeuGc-containing gangliosides. It also reacts with antigens expressed in human breast tumors (Vázquez et al. (1995) Hybridoma , 14, 551-556). In this work, the binding specificity of P3 has been characterized in more detail using a panel of glycolipids that included several disialylated gangliosides and several chemical derivatives of NeuGc-GM3. The carboxyl group and the nitrogen function of sialic acid were found to play important roles in the antibody binding, whereas the glycerol tail appears to be nonrelevant. Molecular modeling was used to analyze the binding data, including the finding that P3 selectively recognizes the internal NeuGc in GD3. For this purpose, conformational studies of GD3 were performed using molecular dynamics. It was concluded that sialic acid binds the P3 antibody through its upper face (the one on which the carboxyl group is exposed) and the C4-C5 side of the sugar ring, whereas none or very little contact between the galactose residue and the protein is evident. Conformational analysis of GD3 revealed that, despite the large flexibility of the NeuGcalpha8NeuGc linkage, the P3 binding epitope on the external sialic acid is not well exposed for any of the possible conformations this linkage can adopt, whereas the internal sialic acid presents the epitope in a proper way for several of these conformations. As a final result, a coherent picture of the epitope that fits the wide binding data was obtained.

Animals↗

T cell-independent B cell response to self-monosialogangliosides: primary response monoclonal antibodies.

Purified GM1 and GM2 gangliosides incorporated into liposomes were injected subcutaneously in BALB/c mice every 3-4 days after pretreatment of the animals with low-dose cyclophosphamide. Serum samples were collected at different intervals and tests by ELISA for the presence of anti-ganglioside antibodies. Four doses (50 micrograms each) were sufficient to raise a measurable primary type of response to GM1, while nine doses were required to obtain measurable IgM antibody titers to GM2. Three monoclonal antibodies (MAbs) wer generated by fusing splenocytes with mouse myeloma cells. The specificity of MAbs was determined by ELISA and HPTLC-immunostaining using a panel of purified glycolipids. The MAb designated E1 showed a high degree of specificity because it reacted only with N-acetyl GM2. Monoclonal antibody A3 reacted predominantly with GM2 and GM1, but also reacted moderately with the GM3 ganglioside. The epitope recognized by this MAb is suggested to be the trisaccharide sequence GalNAc beta 1-4(NeuAc alpha 2-3)Gal. The third MAb (F6) reacted strongly with GM1 but a weak reactivity was also observed with GD1b as well as with asialo-GM1, indicating that the terminal tetrasaccharide Gal beta 1-3GalNAc beta 1-4(NeuAc alpha 2-3)Gal- structure is probably involved in antigenic recognition. Formalin-fixed and paraffin-embedded tissue sections were stained with the E1 and A3 MAbs, using the avidin-biotin complex (ABC) technique. Strong immunoreactivity for E1 appeared in the tumor cells of five primary lung carcinomas and in five malignant melanomas. No immunoreactivity was demonstrated in the parenchyma of a lung without malignancy, or in a metastasis from a colon carcinoma.

Animals↗

Generation of a murine monoclonal antibody specific for N-glycolylneuraminic acid-containing gangliosides that also recognizes sulfated glycolipids.

The P3 murine monoclonal antibody (MAb) was generated by immunizing BALB/c mice with NeuGcGM3 included into liposomes. The specificity of this MAb was defined by an enzyme-linked immunosorbent assay and immunostaining on thin-layer chromatograms. P3 MAb binds to NeuGc-containing gangliosides and was shown also to react with sulfated glycolipids. A preliminary immunohistochemical study showed that the P3 MAb was able to recognize antigens expressed in human breast tumors.

Animals↗

Antigen-specific primary immune response of human B-lymphocytes after in vitro immunization with GM3 ganglioside.

In vitro immunization of human B-lymphocytes was performed with liposomes containing the monosialoganglioside GM3, with or without either complete tetanus toxoid or a synthetic T helper epitope derived from tetanus toxin (determinant 830-843). The immunized B-cells were Epstein-Barr virus transformed and the human anti-ganglioside antibody response was evaluated using an indirect ELISA against different mono- and disialogangliosides. Clones producing antigen-specific human antibodies of the IgM isotype against the ganglioside GM3 used as the immunogen were selected and one clone, IM-11, was further characterized. In addition, a method of positive selection using GM3-coated magnetic beads has been developed which allowed us to rescue unstable clones. The binding of the human antibody IM-11 to a large panel of glycosphingolipids separated on thin-layer plates was studied. The human MAb IM-11 was found to bind strongly to NeuAcGM3, IV3 NeuAcnLc4 and sulfate containing glycosphingolipids and weakly to NeuGcGM3. Immunohistological staining of melanoma and breast cancer biopsy sections showed a selective reactivity of IM-11 with tumor cells which varied among different tumors.

Amino Acid Sequence↗

Somaclonal variation in rye.

We studied the genetic variation generated during in vitro culture of rye Secale cereale L. We analyzed the progenies of four generations of the plants regenerated from immature embryo cultures. A high frequency of mutant plants was observed, 50.75%, this frequency was genotype dependent. Other characteristics typical of somaclonal variation were also observed: the obtaining of dominant mutations, the presence of more than one mutation per plant, the obtaining of homozygous mutants and a high rate of mutation of particular loci. In some cases transposable elements could be implicated. We postulate that tissue culture could induce mutations as well as select particular cell types and so increase the appearance of special mutants.

Crosses, Genetic↗

[Effect of post-natal visit on the involvement in the activities of the mother-child program].

OBJECTIVES: Ascertain what effect post-natal visits have on recruiting involvement in the mother and child programme of activities. Identify the profile of mothers and children with little involvement in these activities. Ascertain how suitable the register is. DESIGN: Study of cohorts of previous years. The term exposed cohort refers to mothers who received the postnatal visit in 1989, and non-exposed cohort to those who did not receive one. SITE. Health Centres in Almanjáyar and Cartuja. Primary Attention. Evaluation of the services offered. PATIENTS OR OTHER PARTICIPANTS: The sample consisted of the 166 women whose pregnancy we monitored from the Centre, and who gave birth between January and December 1989. MAIN MEASUREMENTS AND RESULTS: The most indicative results show that the group that received the post-natal visit tended to attend the healthy child clinic, family planning and metabolism testing services, with greater frequency, and to breast feed longer. The differences are only significant (p = 0.004) in the case of family planning. CONCLUSIONS: We positively evaluate the post-natal visit because of its contribution to the early detection of the problems of mother and child, and to recruitment into Family Planning. And because the examining risk has meant that factors associated with reduced coverage by the services (ethnicity, educational level, mother's age) did not act as such in our study.

Cohort Studies↗

Antitumor properties of an anti-idiotypic monoclonal antibody in relation to N-glycolyl-containing gangliosides.

We examined the antitumor effects of 1E10 monoclonal antibody, an anti-idiotypic IgG to an IgM monoclonal antibody, named P3, that reacts specifically with N-glycolyl-containing gangliosides and also recognizes antigens in human breast and melanoma tumors. Two murine tumor cell lines positive for the P3 antibody, F3II mammary carcinoma (BALB/c) and B16 melanoma (C57BL/6), were employed. In BALB/c mice, vaccination with several i.p. doses at 14-day intervals of 50 microgram of 1E10 coupled to keyhole limpet hemocyanin in Freund's adjuvant, significantly reduced s.c. tumor growth of F3II carcinoma cells and the number of spontaneous lung metastases. Also, the effect of 1E10 as a biological response modifier on tumor lung colonization was evaluated in C57BL/6 mice injected i.v. with B16 melanoma cells. Interestingly, i.v. administration of 10 microgram of uncoupled 1E10 antibody, 10-14 days after inoculation of B16 cells, dramatically reduced the number of experimental metastases in comparison with lungs from mice treated with an irrelevant IgG. The present data suggest that this 'non-internal image' anti-idiotypic monoclonal antibody may activate more than one mechanism of antitumor response against melanoma and mammary tumor cells.

Animals↗