PubMed Health⌕ Search

Biomedical subjects

F Vidal

Publications and source records attributed to F Vidal.

At least 145 records · Page 8Linked to original sources
PubMed1995

Cloning of the laminin alpha 3 chain gene (LAMA3) and identification of a homozygous deletion in a patient with Herlitz junctional epidermolysis bullosa.

Laminin 5 and laminin 6 are basement membrane proteins synthesized by the basal cells of stratifying squamous epithelia. Altered expression of laminin 5 has been associated with Herlitz junctional epidermolysis bullosa (H-JEB), a severe epidermal blistering disorder inherited as an autosomal recessive disease. We have isolated cDNA clones encoding the alpha 3 chain of laminin 5 and searched for mutations in the LAMA3 gene in H-JEB patients. In one H-JEB family, an affected individual exhibited drastically reduced immunoreactivity to antibodies directed against the alpha 3 chain of laminin 5 and an impaired expression of the corresponding mRNA transcripts. RT-PCR analysis of mRNA extracted from the proband's keratinocytes identified a homozygous single basepair deletion in the transcripts encoding the laminin alpha 3A and alpha 3B isoforms. The mutation causes a frameshift and premature termination codon in both alleles of the LAMA3 gene. Inheritance of the clinical H-JEB phenotype was consistent with the segregation of the mutated allele in the family. We also report the identity of the alpha chains of laminin 5 and epiligrin and provide evidence that LAMA3 transcripts are distinct from the laminin 6 alpha chain mRNA.

3T3 Cells↗
PubMed1995

Degradation of fluorescent and radiolabelled sphingomyelins in intact cells by a non-lysosomal pathway.

The aim of the present study was to investigate the role of the entitled neutral, sphingomyelinase in the non-lysosomal pathway of sphingomyelin degradation by intact cells (Spence et al. (1983) J. Biol. Chem. 258, 8595-8600; Levade et al. (1991) J. Biol. Chem. 266, 13519-13529). The uptake and degradation of sphingomyelin by intact living cells was studied using cell lines exhibiting a wide range of activity levels of acid, lysosomal and neutral sphingomyelinases as determined in vitro on cell homogenates by their respective standard assays. For this purpose, neuroblastoma, skin fibroblasts, lymphoid and leukemic cell lines, some of them derived from patients with Niemann-Pick disease (deficient in the acid, lysosomal sphingomyelinase) were incubated with radioactive, [oleoyl-3H]sphingomyelin or fluorescent, pyrene-sulfonylaminoundecanoyl-sphingomyelin. Either compound was taken up by a pathway which was not receptor-mediated and hydrolyzed by all intact cells, including those derived from Niemann-Pick disease patients. Moreover, their degradation by the intact cells was not inhibited by treatment with chloroquine, indicating hydrolysis by a non-lysosomal sphingomyelinase. The intracellular sphingomyelin degradation rates showed no correlation with the activity of the 'classical' neutral sphingomyelinase as determined in vitro. In particular, fibroblasts derived from Niemann-Pick patients lacking the lysosomal sphingomyelinase, and having no detectable in vitro activity of the 'classical' neutral sphingomyelinase, were able to degrade the exogenously supplied sphingomyelins. Indeed, in vitro these cells were shown to exhibit neutral, magnesium- and dithiothreitol-dependent sphingomyelinase activities, that might contribute to the non-lysosomal pathway for sphingomyelin degradation to ceramide in intact cells.

Bridged-Ring Compounds↗
PubMed1995

USF binds to the APB alpha sequence in the promoter of the amyloid beta-protein precursor gene.

The APB alpha domain in the amyloid beta-protein precursor (APP) promoter contains a nuclear factor binding domain with the core recognition sequence TCAGCT-GAC. Proteins in nuclear extracts from brain and numerous cell lines bind to this domain and it contributes approximately 10-30% to the basal APP promoter activity. Included in this domain is the CANNTG motif, which is recognized by basic helix-loop-helix transcription factors. The same motif is also present in the CDEI element of the yeast centromere and in the adenovirus major late promoter (AdMLP). Here we present evidence based on thermostability, relative binding affinity, eletrophoretic mobility and antibody recognition that the cellular proteins that bind to the APB alpha and CDEI motifs are USF. However, the relative binding affinity for the motifs is different. The affinity of USF for AdMLP is approximately 20-fold higher than for the APB alpha sequence and 5-fold higher than for the CDEI sequence. Mutational analysis suggested that the primary determinant for USF binding affinity resides within the octamer CAGCTGAC, which is composed of the E-box consensus sequence CANNTG followed by the dinucleotide AC. The human homolog of the mouse CDEI binding protein did not bind to either the CDEI sequence or APB alpha.

Adenoviridae↗
PubMed1995

Programming the duration of a motor sequence: role of the primary and supplementary motor areas in man.

Event-related potentials were recorded in a reaction time (RT) paradigm, where the duration of a learned interval (either 0.7 s or 2.5 s) delimited by two brief button-presses was to be accurately controlled. A preparatory signal (PS) either did not give or gave prior information concerning the duration of the following response (neutral condition or primed conditions, respectively). In the latter case, the information was either validated (valid condition) or invalidated (invalid condition) by the response signal (RS). When duration was not known in advance (invalid and neutral conditions), RTs were longer before a response of short than long duration. This difference was not found under the valid condition. During the preparatory period (PP), the amplitude of the contingent negative variation (CNV) was larger when the duration was primed than when it was not. A larger CNV appeared when the PS primed a short rather than a long duration. This effect occurred in the early part of the PP over the supplementary motor area (SMA) and in its latest part over the primary motor area (MI). The RT and the electrophysiological pattern were interpreted as revealing the occurrence of programming operations regarding the temporal dimension of the response. The time course of the CNV over the SMA and MI suggested that these two areas were hierarchically organized. Between the RS and the onset of the response, differences probably related to programming effects were still found over MI: the activities were larger under the valid than under the neutral condition. However, no sign of deprogramming (expected in the invalid condition) was observed: similar amplitudes were found under the neutral and invalid conditions. Deprogramming operations seemed to be postponed during response execution where the invalid condition evoked larger activities than the two other conditions over the SMA. Finally, MI but not the SMA yielded a Bereitschaftpotential before the second press ending the response (i.e., during response execution). These results suggest that the duration of a motor response can be a part of the motor program and that the SMA plays a major role in programming processes but not in response execution, contrary to MI.

Algorithms↗
PubMed1995

Cytogenetic studies of oocyte fusion products.

We describe for the first time the cytogenetic characteristics of mouse 'embryos' obtained by oocyte fusion (oocyte fusion products; OFP). Our results indicate that, after fusion, meiosis II is resumed correctly, with extrusion of two haploid polar bodies, and that metaphase synchronisation of the two haploid sets and chromosome segregation during the first cleavage are also normal.

Animals↗
PubMed1995

Early mouse preimplantation development is unaffected by microinjection of metallothionein antibodies.

Polyclonal antibodies that cross-react with rodent metallothionein I (MT I) and metallothionein II (MT II) were microinjected in 1-cell and 2-cell mouse embryos, into either the cytoplasm or the nucleus. Regardless of the experimental treatment, mouse embryo development in vitro was not affected and most of the embryos cleaved normally until the morula stage. The results suggest that metallothionein is not essential for normal mouse early preimplantational development, in agreement with recent studies in mice with inactivated MT I and MT II genes.

Animals↗
PubMed1995

Integrin beta 4 mutations associated with junctional epidermolysis bullosa with pyloric atresia.

Pyloric atresia associated with junctional epidermolysis bullosa (PA-JEB), is a rare inherited disorder characterized by pyloric stenosis and blistering of the skin as primary manifestations. We demonstrate that in one PA-JEB patient the disease resulted from two distinct mutations in the beta 4 integrin gene alleles. The paternal mutation consists of a one base pair deletion causing a shift in the open reading frame, and a downstream premature termination codon. The maternal mutation occurs in a donor splice site, and results in in-frame exon skipping involving the cytoplasmic domain of the polypeptide. Our results implicate mutations in the beta 4 integrin gene in some forms of PA-JEB.

Alleles↗
PubMed1995

CDEBP, a site-specific DNA-binding protein of the 'APP-like' family, is required during the early development of the mouse.

A murine protein, termed CDEBP, was previously shown to bind the double-stranded DNA motif GTCACATG, identical to the yeast centromeric element CDEI. The cDNA sequence showed three domains with extensive similarities to the amyloid beta precursor protein (APP). The protein is homologous over its entire length to the human protein designated APPH. In situ immunofluorescence assays using antibodies raised against distinct parts of CDEBP detected discrete sites of accumulation inside the interphase nucleus, and the bulk of the protein was not associated with mitotic chromosomes. One of the complexes with double-stranded CDEI oligonucleotides detected by gel shift assay was not present when the protein had been selectively removed from nuclear extracts by immunoprecipitation. We reported previously that microinjection into one-cell mouse embryos of DNA fragments including the CDEI sequence results in an early arrest of development with abnormal nuclei containing variable amounts of DNA. The same characteristic figures were observed when embryos were treated with antisense oligonucleotides complementary to parts of the CDEBP coding region. Complexes between the CDEBP protein and CDEI sites in the mouse genome thus appear to play a critical role in the replication/segregation of the embryonic genome.

3T3 Cells↗
PubMed1995

[Evaluation of serum adenosine deaminase as a prognostic marker in the treatment of human immunodeficiency virus infection with zidovudine].

BACKGROUND: Adenosine deaminase (ADA) is a key enzyme in cellular immunity. It catalyses the reaction 2'deoxyadenosine to 2'deoxiinosine, a critical step in the production of essential metabolites for the synthesis of nucleic acids. Its main physiological activity occurs in T cells of lymphoid tissue. The advanced HIV infection is characterized by a severe and progressive cellular immunity compromise. Partial positive results have been obtained with Zidovudine (AZT), a drug which Serum adenosine deaminase has been proposed as a useful marker for the assessment of the therapy response in these patients. METHODS: Serum ADA activity was determined in patients infected with HIV in stage II, before and after four months of therapy with AZT, regarding variations in its activity with improvements in biological parameters--CD4 and CD8 lymphocytes. RESULTS: Patients infected with HIV showed a significant increase in ADA activity compared with patients in the control group: 21.6 +/- 5.4 vs. 10.4 +/- 2.3 U/l (p < 0.001). Therapy with AZT decreased ADA activity: 21.6 +/- 5.4 vs. 15.2 +/- 4.3 U/l (p < 0.001) and correlated with an increase in CD4 counts: 187 +/- 105 vs. 353 +/- 145/mm3 (p < 0.001) and in CD4/CD8 ratio: 0.188 +/- 0.10 vs. 0.382 +/- 0.18 (p < 0.001). CONCLUSIONS: The detection of a decrease in ADA correlates significantly with a favourable outcome in immunological parameters in individuals infected with HIV in stage II of disease treated with AZT.

Adenosine Deaminase↗
PubMed1995

Evaluation of cytogenetic analysis for clinical preimplantation diagnosis.

OBJECTIVE: To evaluate the feasibility of using cytogenetic analysis in preimplantation diagnosis. DESIGN: Two different biopsy protocols (chemical drilling and zona cutting) and two fixation methods were tested in a mouse model. Afterwards, the efficiency of obtaining chromosome preparations from untransferable human embryos depending on the method used to obtain the blastomeres (embryos biopsy or removal of the zona pellucida and blastomere disaggregation) was determined. The chances of obtaining chromosome preparations depending on the type of embryo (haploid, diploid, triploid, and apparently unfertilized) were also evaluated. RESULTS: Results from the mouse model showed that chemical drilling yields better results than cutting in terms of metaphases per biopsied embryo and surviving rate after biopsy. In human embryos, biopsy of diploid embryos produced 46.6% chromosome preparations, while 29% were obtained after blastomere disaggregation and 20.4% when biopsying triploid embryos. CONCLUSIONS: These results suggest that the disaggregating procedure and triploid embryos cannot be considered as good models to assess the feasibility of cytogenetic analysis in preimplantation diagnosis. Poor chromosome quality and loss during fixation are the main problems to use cytogenetics in preimplantation diagnosis; a combination of cytogenetics and other techniques is suggested in cases of balanced translocations.

Animals↗
PubMed1995

Cell biology of restenosis post-angioplasty.

PTCA is a well established intervention to reduce the severity of atherosclerotic coronary stenosis. In spite of a primary success rate of 90 - 95%, late restenosis occurs in 30 - 50% of patients within 3/6 months of the procedure. Angioplasty in swine induces similar events to those found in humans, thus providing a model for studying strategies for intervention. Blood interaction to the damaged vessel wall occurs with reperfusion after the intervention. Therefore, the in vivo characterization of the interaction of cellular elements (platelets and white cells) and blood proteins with the exposed vascular cells in the vessel wall post-angioplasty may be necessary to identify early triggers of restenosis. Angioplasty was performed simultaneously in the coronary and carotid arteries of swine by fluoroscopy assisted standard techniques. Angiography was performed acutely post-dilatation and residual lumen diameter evaluated. Dilated vessels from 30 min to 6 h postintervention were processed to prepare RNA and preserved to perform immunohistochemistry. Dilatation injury induces maximal expression of c-fos 30 min and c-myc from 2 to 4 h postdilatation. Platelet deposition is initiated immediately post-dilatation as well as infiltration of fluid phase proteins on the damaged areas.

Angioplasty, Balloon, Coronary↗
PubMed1995

Peribulbar anaesthesia: the role of local anaesthetic volumes and Thiomucase in motor block and intraocular pressure.

Peribulbar anaesthesia is a useful regional anaesthesia and akinesia technique for eye surgery. We studied changes in muscular akinesia and intraocular pressure under peribulbar anaesthesia caused by different volumes of 0.75% bupivacaine with or without the addition of Thiomucase (chondroitin sulphatase) in patients undergoing cataract surgery. A total of 120 patients were randomly allocated into four groups: group 1, bupivacaine 5 mL; group 2, bupivacaine 5 mL plus Thiomucase 10 U mL-1; group 3, bupivacaine 9 mL; group 4, bupivacaine 9 mL plus Thiomucase 10 U mL-1. We assessed: (1) akinesia according to mobility in the four quadrants of the eyeball and the palpebral opening using a scoring system between 0 (total akinesia) and 10 (full movement); and (2) intraocular pressure before, immediately after and 15 min after peribulbar anaesthesia. A significantly better motor blockade was observed in groups 2, 3 and 4 than in group 1 (P < 0.05). Thiomucase increased the quality of motor blockade in group 2, but not in group 4. Intraocular pressure increased by 5-10 mmHg in comparison with baseline values and, after 15 min application of Honan's balloon, decreased by 8-10 mmHg in comparison with baseline values, with no significant differences between groups. Thiomucase can improve the motor block if a lower volume of local anaesthetic is used. The administration of 5 or 9 mL volumes or the addition of Thiomucase decreases the the intraocular pressure in a similar way.

Aged↗
PubMed1994

Difficult management of pain following sacrococcygeal chordoma: 13 months of subarachnoid infusion.

We report on a patient suffering severe pain following a long-standing sacral chordoma in whom management of therapy and pain was extremely difficult. Because orally administered morphine was observed to be ineffective in the early stages of treatment, we tried to achieve pain relief by using epidural morphine. This was also unsatisfactory. Intrathecal infusion of morphine and bupivacaine through a catheter inserted at the L2-L3 level was also found to be ineffective; only a segment block was observed. A structural anomaly of the spine was suspected and confirmed by myelography, showing metastasis at L3. Pain relief improved when an infusion of morphine+bupivacaine was employed through a catheter placed at the L4-L5 level below the blockage. A major problem which continued throughout the course of the disease was the daily occurrence of episodes of unbearable sharp pain that required addition of midazolam to the local anesthetic/opioid subarachnoid infusion. The patient received this mixture through an intrathecal port during the last 13 months of life, a gradually increasing dose being necessary. Periods of analgesia were followed by occasional crises of intense sharp pain suggesting incomplete relief. No serious complications or meningitis occurred. This case emphasizes the difficulty in managing pain in this type of cancer.

Bupivacaine↗
PubMed1994

Efficacy and tolerance of oral dipyrone versus oral morphine for cancer pain.

In a double-blind, randomised and parallel clinical trial, two oral doses of dipyrone (1 and 2 g) administered every 8 h were compared with 10 mg of oral morphine given every 4 h for the relief of chronic cancer pain. A total of 121 patients with cancer pain without gastric involvement participated in a 7-day treatment course and were allocated to receive either dipyrone 1 g (n = 41), dipyrone 2 g (n = 38) or morphine (n = 42). Drug efficacy was analysed according to the degree of pain relief using a 100-mm visual analogue scale, and the number of patients who decided to increase the dose of the analgesic drug on day 4. The analgesic effect of dipyrone, 2 g every 8 h, was similar to that of morphine. The efficacy of both schedules was significantly greater than that of dipyrone, 1 g every 8 h. Dipyrone at either 1 or 2 g doses tended to be better tolerated than morphine, although the differences were not statistically significant.

Adult↗
PubMed1994

Ultrastructural studies of early mouse embryos obtained by oocyte fusion.

Oocyte fusion induced by inactivated Sendai virus results in the production of 'zygotes' that are able to undergo the first stages of embryonic development. The oocyte fusion products (OFP) obtained follow a morphological developmental pattern equivalent to that of control embryos, at least up to the 8-cell stage. The percentage of OFP that reach the 8-cell stage is extremely low (3%) compared with control embryos cultured in vitro (95%). On light microscopy, the OFP obtained show morphological characteristics identical to control embryos, although their cell diameters are larger. The cortical reaction, meiotic reactivation, extrusion of second polar bodies and pronucleus formation take place as observed in controls. The ultrastructural characteristics of oocyte fusion products at the 1-, 2-, 4- and 8-cell stages are analogous to those of controls, including the presence of structures related to the activation of the embryo genome. However some differences concerning cell ultrastructure, mainly in the nucleus, are observed and discussed in the text.

Animals↗