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Biomedical subjects

M Martí

Publications and source records attributed to M Martí.

At least 19 recordsLinked to original sources

Characteristics of actin fibers and ultrastructure of the contact regions involved in the separation of blastomeres of two-cell mouse embryos, frozen-thawed without the zona pellucida.

Freezing of embryos deprived of the zona pellucida (ZP) decreases their survival rate immediately after thawing, and gives rise to the separation of their blastomeres in a high percentage of cases. We have studied the ultrastructure and the characteristics of actin fibers in the cell-to-cell contact region in mouse embryos frozen-thawed without the ZP at the two-cell stage. Our results indicate that most of the embryos that retain their blastomeres united after freezing and thawing show either the presence of a midbody, or a contact region with a close apposition of the plasma membranes but without an organized actin cortex in their contact region. Only a small percentage of embryos that retain their blastomeres united after freezing and thawing show a contact region with widely separated plasma membranes and an organized actin cytocortex.

Actins

Inhibition of DNA synthesis by aspirin in Swiss 3T3 fibroblasts.

In Swiss 3T3 fibroblasts, aspirin inhibited proliferation induced by the complete mitogenic factors platelet-derived growth factor (PDGF) and bombesin. Aspirin decreased the maximum mitogenic effect of bombesin without modifying the concentration necessary to obtain half maximal DNA synthesis stimulation. In contrast, aspirin only decreased mitogenesis at subsaturating PDGF concentrations. The effect of aspirin was found to be concentration dependent. The half-maximal effect occurred at approximately 150 microM. The maximal inhibition was obtained when aspirin was added during the first hour after growth factor addition. At this time, both PDGF and bombesin induced prostaglandin E2 synthesis. PDGF induced much higher levels of prostaglandin E2 than bombesin. The inhibitory effects of aspirin on PDGF or bombesin-stimulated DNA synthesis were counteracted by 280 nM prostaglandin E2. Aspirin effects were overcome by agents that increase cellular cyclic adenosine monophosphate levels but not by activation of protein kinase C. The significance of the antiproliferative action of aspirin might be associated with epidemiological data that show a reduced incidence of colorectal and other cancers after aspirin treatment.

3T3 Cells

Advantages of using a cell separator and metrizamide gradients for human islet purification.

Human islet transplantation has a high rate of failure, often due to primary nonfunction, which suggests that islets are damaged during the processing of the pancreas. The preparation of human islets for transplantation is still a complex process that requires large teams of surgical and laboratory personnel. To overcome this problem, we have adopted the use of the IBM 2991 COBE cell separator and a metrizamide/Ficoll density medium that is easy to prepare. Twenty-seven pancreatic glands have been processed using the COBE cell separator, 23 of which were purified in metrizamide/Ficoll gradients and 4 in bovine serum albumin gradients. The results show an improvement of recovery and viability in these preparations when compared retrospectively with manual gradients. More importantly, the time required for purification was shortened to one fourth the usual time and total processing time is about half as long. Moreover, a team of two laboratory staff was regularly able to prepare islets for transplantation, reducing the separation time from 7 hr to 3.5 hr. We conclude that the automatic cell separator and metrizamide-based separation medium are useful modifications of current islet purification methods.

Adolescent

[Women and breast feeding: understanding their experiences and analyzing the performance of the health system].

OBJECTIVE: To understand the conditioning factors which, whether negatively or positively, affect the success or failure of breast-feeding and to analyse the intervention of the health system to support it. DESIGN: A descriptive, prospective study which uses qualitative methodology in data gathering and analysis. SETTING: Health District of Santa Perpetua de la Mogoda, Barcelona. PARTICIPANTS: 56 in-depth interviews of 20 women were carried out at different moments of the pregnancy and suckling. MEASUREMENTS AND MAIN RESULTS: The decision to breast-feed was taken by women before or during their pregnancy under the influence of their social and family environment. The performance of the health system had little influence on the decision. The birth, even if it was pathological, did not prevent breast-feeding except when it was experienced as a failure. The days before the milk came increased women's insecurity about their capacity to suckle. Information from hospitals was evaluated as being scant. Fear of not feeding the baby well was the major source of breast-feeding failure. Mothers who breast-fed for longer felt they received more sympathy from paediatricians than those who gave up early. These felt blamed and did not seek doctors' advice on their decision. CONCLUSIONS: The health system has to plan an efficient intervention to support breast-feeding in the most crucial moments: after the birth and on arriving home.

Adult

Ultrastructural study of turkey rhinotracheitis virus infection in turbinates of experimentally infected chickens.

Ultrastructural changes associated with turkey rhinotracheitis virus infection were studied in turbinates of chickens experimentally infected with the isolate CVL 14/86/1. Chickens were sacrificed at 3, 5 and 7 days after inoculation and samples of the middle turbinate were taken, fixed, dehydrated and embedded in an hydrophilic resin. An immunofluorescence technique on semithin sections was carried out and viral antigen was observed in the cytoplasm and associated to cilia of the turbinate epithelial cells, on days 3 and 5 after inoculation. Ultrastructurally, gold stained intracytoplasmic nucleocapsid aggregates of turkey rhinotracheitis virus were observed in ciliated and non-ciliated epithelial cells, as well as budding virus particles, at days 3 and 5 postinoculation. Different ultrastructural abnormalities, including cytoplasmic blebs, clumping and loss of cilia were observed in the apical cell membrane of many infected cells, associated with the presence of intracytoplasmic inclusions. On day 5 after inoculation, substitution of ciliated and non-ciliated epithelial cells was noted and many desquamated epithelial cells were observed within the lumina. Regenerative changes in the ciliated epithelium were observed by day 7 postinoculation. These results indicate that turkey rhinotracheitis virus is able to replicate in ciliated and non-ciliated epithelial cells causing severe alterations to the cell surface and ciliary apparatus of the turbinate epithelium. Viral-induced damage to the turbinate epithelium could enhance the susceptibility of epithelial cells to secondary bacterial infection.

Animals

Freezing of zona-free mouse embryos: characteristics of the plasma membrane and subsequent development of the embryos.

Frozen-thawed mouse embryos with (+ZP) and without (-ZP) zona pellucida have been studied at the Scanning Electron Microscope (SEM) to determine how the process affects the plasma membrane and the subsequent embryo development. The main difference observed in -ZP embryos immediately after thawing is the abnormal morphology and distribution of microvilli. This could explain the spontaneous separation of blastomeres in -ZP embryos, and the decrease in their survival rate. If thawed -ZP embryos are allowed to recover in culture, their plasma membrane characteristics and survival rate are identical to those of control embryos.

Animals

Pancreas in recent onset insulin-dependent diabetes mellitus. Changes in HLA, adhesion molecules and autoantigens, restricted T cell receptor V beta usage, and cytokine profile.

Insulin-dependent diabetes mellitus (IDDM), in which only the pancreatic beta cells are destroyed by the autoimmune response, is the paradigm of organ-specific autoimmunity. As a result of a combination of factors, the number of immunohistologic/cellular/molecular studies of pancreas in IDDM is very limited. We report here studies conducted in the pancreata of two IDDM patients: one newly diagnosed (case 1) and one long standing (case 2). In case 1, we demonstrated the presence of morphologically normal viable beta cells without evidence of viral infection. In both cases the expression of the autoantigens defined by islet cell Abs and by glutamic acid decarboxylase was markedly reduced in the islet cells whereas expression of hsp60, another putative autoantigen, was normal. Over-expression of HLA class I was detected in 58% of the islets in pancreatic sections and in cultured beta cells in case 1 and also in 30% of islets in case 2 but it was not restricted to any insular cell type. In case 1, there was "inappropriate" HLA class II expression in islets cells but it was a rare finding and not beta cell specific. The analysis of the correlation between class I overexpression, residual insulin, and insulitis suggests that the first event is the increase of HLA class I expression. Of adhesion molecules, ICAM-1, VLA, VCAM, and LFA-3 were normal and only ICAM-1 was moderately overexpressed in and around the islets of case 1 insulitis, as was detected by immunofluorescence which showed that 18% of the islets of case 1 had CD8+ lymphocytes as the predominant population. Reverse transcription-PCR demonstrated moderate V beta skewing and the profile of cytokines expected in CTLs: IL-2, IL-4, IL-10, and IFN-gamma negative, perforin positive. In addition, IFN-alpha, IFN-beta, and IL-6 transcripts were detected in the case 1 pancreas, consistent with the existence of a silent viral infection. Overall, the results indicated that, differently from spontaneous animal models of diabetes, in the pancreas of IDDM patients there are no elements of the inductive phase of the autoimmune response.

Acute Disease

Different requirements of ICAM-1/LFA-1 adhesion in allorecognition and self-restricted antigen recognition by class II-specific T cell clones.

We have analyzed the influence of non-antigen-specific interactions between ICAM-1 and LFA-1 in target recognition by allospecific and antigen-specific T cells at the clonal level, using human and mouse fibroblasts transfected with HLA-DR1 or DR2 with or without co-expression of ICAM-1, as antigen-presenting cells. The results show a great heterogeneity in the requirements for ICAM-1/LFA-1 interactions for antigen-specific and alloreactive T cell responses and this requirement may depend on the avidity of any particular interaction. The data also show that for most alloreactive clones, ICAM-1/LFA-1 adhesion is not sufficient to facilitate efficient T cell recognition of its target molecule. HLA class II recognition by a large proportion of the DR1- and DR2-specific alloreactive clones studied was different for class II molecules expressed on murine or human fibroblasts compared to human lymphoid cells, and was independent of ICAM-1 expression on the stimulator cells. The inability of some T cell clones to recognize HLA-class II expressed on non-lymphoid cells suggests the absence of specific epitopes and could be due to the lack of the relevant peptides, either because they are derived from species-specific proteins or to differences in processing of endogenous antigen in the transfected stimulator cells.

Animals

A simple method for processing individual oocytes and embryos for electron microscopy.

A simple method for handling individual specimens that must be processed either for scanning or transmission electron microscopy studies is described. For scanning microscope processing, dehydration is carried out with samples enclosed in small cages made from TAAB capsules in which top and bottom are substituted by plankton nets, and for transmission electron microscopy, samples are preembedded in agarose. This procedure significantly reduces mouth pipetting, dissecting microscope observations, is less labour intensive and, most importantly, reduces sample loss.

Animals

[Paraplegia following surgical repair of a ductus and of a coarctation of the aorta in childhood].

The cases of two girls with 4 and 7 years of age presented. They had a patent ductus and an aortic coarcation which were subsequently surgically repaired. After the operation paraplegia was diagnosed. With rehabilitation they are able to walk, however paraparexis persist. Anatomy of the arterial irrigation of the spinal cord is reviewed and also the pathophysiology and mechanisms of medullar ischemia.

Aorta, Thoracic

Expression of intercellular adhesion molecule-1 in thyroid follicular cells in autoimmune, non-autoimmune and neoplastic diseases of the thyroid gland: discordance with HLA.

The presence of intercellular adhesion molecule-1 (ICAM-1) on epithelial cells facilitates their recognition by specific T lymphocytes. To assess the possible role of ICAM-1 in the recognition of thyroid follicular cells by T cells in thyroid autoimmune disease, we investigated the expression of ICAM-1 in thyrocytes from thyroid glands affected by Graves' disease, in glands with non-autoimmune pathology and normal glands using immunofluorescence staining on cryostat sections and on dispersed cell preparations. Sequential tissue sections from glands affected by Graves' disease (n = 15), multinodular goitre (MNG, n = 26), benign nodules (n = 11), primary carcinomas (n = 12) and control thyroid glands (n = 5) were stained for ICAM-1, HLA class I, HLA class II, CD3 and thyroid peroxidase (TPO). Weak and patchy ICAM-1 expression was found in the thyrocytes of 4/15 (27%) Graves' disease and of 1/26 (4%) multinodular goitre glands. In contrast, ICAM-1 expression was detected in the thyrocytes of 5/11 (45%) benign nodules and of 8/12 (67%) thyroid carcinomas in which it was sometimes strong. Thyrocytes in the five control glands were negative. These results correlated well with flow cytometry data from 23 of these glands which showed that ICAM-1 expression in thyrocytes from Graves' patients was, when present, 'dull', while in some malignant thyrocytes it was 'bright'. In preparations of thyrocytes from Graves' disease glands we found a striking discordance between the high levels of expression of HLA class I and HLA class II and the low expression of ICAM-1. This is surprising since in vitro the expression of these three molecules is equally induced by IFN-gamma and TNF-alpha. These results suggest that additional factors are involved in the induction of the inappropriate HLA class II expression observed in the thyrocytes of glands affected by Graves' disease.

Antigens, CD

Induction of intercellular adhesion molecule-1 but not of lymphocyte function-associated antigen-3 in thyroid follicular cells.

We investigated the expression of intercellular adhesion molecule-1 (ICAM-1) and lymphocyte function-associated antigen-3 (LFA-3) by thyrocytes and their regulation by cytokines. Immunofluorescence studies on cryostat sections and on freshly dispersed cell preparations showed that ICAM-1 and LFA-3 are barely detectable in non-autoimmune thyrocytes. However, thyrocytes acquired ICAM-1 expression in culture. IFN-gamma, IL-1 beta and TNF-alpha produced a clear enhancement of ICAM-1 expression. When tested in combination, IL-1 beta and TNF-alpha were additive to the IFN-gamma effect. LFA-3 expression was not modulated by these cytokines. In the HT93 thyroid cell line generated by transfection with SV40, ICAM-1 and LFA-3 were both constitutively expressed at high levels. Cytokines modulated ICAM-1 expression similarly, but to a greater extent than in normal thyrocytes. LFA-3 remained unmodified. These results support the notion that normal thyrocytes are immunologically silent cells. The capability of cytokines to induce ICAM-1 together with HLA class I and class II-expression on thyrocytes suggests that under their influence, these cells may express all the surface molecules required for antigen presentation and/or for being recognized as target cells in the context of thyroid autoimmune disease.

Antibodies, Monoclonal