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Biomedical subjects

A Lopez

Publications and source records attributed to A Lopez.

At least 127 records · Page 7Linked to original sources

Efficacy and safety of avocado/soybean unsaponifiables in the treatment of symptomatic osteoarthritis of the knee and hip. A prospective, multicenter, three-month, randomized, double-blind, placebo-controlled trial.

One of the objectives of symptomatic slow-acting drugs for osteoarthritis is to reduce the need for drugs with a less favorable safety profile, mainly analgesics and nonsteroidal antiinflammatory drugs (NSAIDs). We conducted a three-month, prospective, randomized, double-blind, placebo-controlled, parallel-group trial to evaluate the efficacy of avocado/soybean unsaponifiables in terms of NSAID use reduction. Patients with primary femorotibial or hip osteoarthritis (OA) (ACR criteria and Kellgren-Lawrence radiological stage IB, II, or III) of at least six months' duration with regular pain for more than three months requiring therapy with NSAID (visual analog scale score > or = 25 mm and Lequesne's index on therapy > or = 4) were given one capsule per day of avocado/soybean unsaponifiables or a placebo for three months. During the first 45 days, patients in both groups were also given one of seven predefined NSAIDs. The primary efficacy criteria was the rate of patients taking back a NSAID and the delay before re-intake. Secondary efficacy criteria were the total dose of NSAID, overall ratings by the patient and by the physician, the visual analog scale pain score and the functional index. Of the 164 included patients, 163 were evaluable, 80 in the active drug group and 83 in the placebo group. Mean age was 62.9 +/- 8.8 years. The diagnosis was femorotibial OA in 101 patients and hip OA in 62. Data were collected on day 45 in 153 patients (77 on the active drug and 76 on the placebo). The number of patients who took back NSAID therapy was significantly smaller in the group treated by avocado/soybean unsaponifiables (33; 43.4%) than in the placebo group (53; 69.7%) (P < 0.001). Also, beyond day 54, the time spent off NSAID therapy was shorter in the placebo group. The functional index showed a significantly greater improvement in the active drug group (-2.3 +/- 2.6) than in the placebo group (-1.0 +/- 2.6) (P < 0.01). Pain scores over time were similar in the two groups. Overall patient ratings were significantly better in the active drug group (P < 0.01). Safety was oggd in both groups. After six weeks, avocado/soybean unsaponifiables reduced the need for NSAID in patients with lower limb OA.

Aged↗

Multiparametric cell-cycle analysis of peripheral blood-activated lymphocyte subsets using staining based on the TdT method for incorporated BrdUrd.

This study describes a new method for the simultaneous assessment of the distribution of a cell population in the G0/G1, S, and G2/M cell-cycle phases by using multiparameter flow cytometry and single staining based on BrdUrd incorporation. Both the K562 cell line and PHA-stimulated peripheral blood lymphocytes (PBL) were analyzed. Cells were cultured in the presence of BrdUrd for 30 min prior to cell harvesting. Once collected, cells were exposed to ultraviolet light for 5 min and then fixed immediately in 70% ethanol (-20 degrees C) for at least 30 min. Once fixed, the cells were placed for 30 min at 37 degrees C in the presence of terminal deoxynucleotidyl transferase (TdT) and dUTP labeled with digoxigenin; they were then stained with FITC-labeled anti-digoxigenin. Our results show that G0/G1, s, and G2/M cell populations can be clearly discriminated according to FITC fluorescence and light-scatter parameters. In this way, S-phase cells can be identified by their FITC staining. From the cells which were negative for anti-digoxigenin-FITC antibody, two clear populations could be resolved in a forward scatter, side scatter, and fluorescence pulse-width three-dimensional plot; the values obtained for G0/G1 cells were lower than those obtained for G2/M cells in all three parameters. Multiparameter analysis of PBL stained for two surface antigens (CD3 and CD8) and for BrdUrd by direct or indirect TdT method permitted cell-cycle analysis of different subpopulations, including CD3+/CD8+, CD3+/CD8-, CD3-/CD8+, and CD3-/CD8-.

Bromodeoxyuridine↗

Comparison of Vindelov et al. and bromodeoxyuridine/DNA double-staining flow cytometry methods for analysis of cell cycle distribution in rat thymocytes.

This study compares the cell cycle distribution in rat thymocytes obtained by means of bromodeoxyuridine (BrdUrd) labeling of S-phase cells and the analysis of the S-phase fraction obtained according to the technique of Vindelov et al. (Cytometry 3:332-338, 1983). The proportion of BrdUrd-labeled cells was analyzed in single cell suspensions of adult rat thymocytes after in vivo injection of BrdUrd and the results then compared with those obtained after measuring the cell DNA contents according to the Vindelov et al. method. The percentage of BrdUrd-positive cells was greater than the S-phase fraction obtained using the Vindelov et al. technique. By contrast, no major differences were observed between the percentage of BrdUrd-positive cells and the S-phase fraction obtained after analyzing the DNA histograms of the same data files with the RFIT mathematical model. The elimination of trypsin treatment used in the Vindelov et al. method did not alter the results, whereas the use of DNA denaturation with 2N HCl was shown to increase the percentage of S-phase rat thymocytes (calculated from DNA histograms) independently of whether trypsin treatment was used or not. However, the value of the S-phase fraction was not as great as that obtained after BrdUrd labeling. Thus when comparing BrdUrd-labeling and the Vindelov et al. technique, important differences in the percentage of S-phase adult rat thymocytes were observed. Selective G0/G1 cell loss during washing and centrifugation steps performed after the DNA denaturation used for BrdUrd detection was the main reason for these differences.

Animals↗

Strandedness discrimination in peptide-polynucleotide complexes.

Preferential binding to single- or double-stranded nucleic acids is important for the activity of many proteins that process RNA and DNA. We have investigated the mechanism of strandedness discrimination with peptides derived from the putative DNA-binding domain of the RecA protein, a bacterial recombinase that modulates its affinity for single-stranded DNA by means of ATP binding and hydrolysis. Contributions of electrostatic and non-electrostatic interactions to binding of these peptides with polynucleotides were evaluated by fluorescence spectroscopy as a function of salt concentration and peptide charge. Binding of these peptides to single- and double-stranded nucleic acids was dominated by non-electrostatic interactions. Small electrostatic contributions selectively enhanced peptide complexation with single-stranded nucleic acids. Similar results were observed in control experiments carried out with tripeptides containing charged and aromatic amino acid residues. It was possible to modify the strandedness preference of peptide-polynucleotide complexes by changing electrostatic contributions to the binding free energy. These observations suggest a mechanism whereby some proteins that interact with DNA or RNA might determine and regulate their relative affinity for single- and double-stranded nucleic acids.

Amino Acid Sequence↗

Granulocyte-macrophage colony-stimulating factor receptor: stage-specific expression and function on late B cells.

Granulocyte-macrophage colony-stimulating factor (GM-CSF) receptors (GMR) are expressed on myeloid cells throughout their maturational sequence. During myelopoiesis, GM-CSF induces the proliferation of precursors and has multiple effects on more mature cells; such effects include induction of maturation and priming for subsequent stimulation. GMR is expressed on a range of other cell types including acute leukemic blasts of myeloid and lymphoid lineage, but has been little studied on more mature lymphoid cells. Using sensitive triple-layer immunophenotypic techniques, we show here that both the alpha and beta c chains of the GMR are expressed on hairy cells (HCs) and myelomatous plasma cells (PCs), but not on chronic lymphocytic leukemia (CLL) or prolymphocytic leukemia (PLL) lymphocytes. The receptor was demonstrable on normal PCs in tonsil, but not on either activated or resting tonsillar B cells or on circulating normal B lymphocytes. The expression of the receptor is therefore stage specific, rather than a feature of activation. Perhaps, surprisingly, in view of its effects on myeloid cells, GM-CSF did not stimulate the proliferation or differentiation of HCs and did not protect them from apoptosis. However, the cytokine had a profound effect on the interaction of the HC with its environment. Thus, the cytokine caused a major cytoskeletal reorganization resulting in the inhibition of motility and loss of adhesion to cellular and matrix ligands. These studies indicate the importance of GM-CSF outside myelopoiesis and demonstrate a previously unrecognized stage specific role for the cytokine in B-cell biology. Taken together with our previous report that M-CSF enhances B-cell motility, the present findings indicate that myeloid growth factors act in concert to facilitate the controlled migration of certain B cells into and within tissues.

Apoptosis↗

7-nitrobenz-2-oxa-1,3-diazole-4-yl-labeled phospholipids in lipid membranes: differences in fluorescence behavior.

Steady-state and time-resolved fluorescence properties of the 7-nitrobenz-2-oxa-1, 3-diazole-4-yl (NBD) fluorophore attached either to the sn-2 acyl chain of various phospholipids (phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and phosphatidic acid) or to the polar headgroup of phosphatidylethanolamine were studied after insertion of these NBD-labeled lipid probes into unilamellar vesicles of phosphatidylcholine, phosphatidylglycerol, phosphatidic acid, and phosphatidylserine. The fluorescence response of the NBD group was observed to strongly depend on the chemical structure and physical state of the host phospholipids and on the chemical structure of the lipid probe itself. Among the various fluorescence parameters studied, i.e., Stokes' shifts, lifetimes, and quantum yields, the quantum yields were by far the most affected by these structural and environmental factors, whereas the Stokes' shifts were practically unaffected. Thus, depending on the phospholipid probe and the host phospholipid, the fluorescence emission of the NBD group was found to vary by a factor of up to 5. Careful analysis of the data shows that for the various couples of probe and host lipid molecules studied, deexcitation of the fluorophore was dominated by nonradiative deactivation processes. This great sensitivity of the NBD group to environmental factors originates from its well-known solvatochromic properties, and comparison of these knr values with those obtained for n-propylamino-NBD in a set of organic solvents covering a large scale of polarity indicates that in phospholipids, the NBD fluorophore experiences a dielectric constant of around 27-41, corresponding to a medium of relatively high polarity. From these epsilon values and on the basis of models of the dielectric transition that characterizes any water-phospholipid interface, it can be inferred that for all of the phospholipid probes and host phospholipids tested, the NBD group is located in the region of the polar headgroups, near the phosphoglycerol moiety of the lipids.

1,2-Dipalmitoylphosphatidylcholine↗

Clinical and microbiological characteristics of Rahnella aquatilis strains isolated from children.

We report the microbiological characteristics of two Rahnella aquatilis strains isolated in the faeces of two patients with acute gastroenteritis, one of whom is an AIDS patient. The biochemical behaviour was studied with different automated identification systems, and the few clinical cases to be found in the literature were reviewed. Of the nine strains isolated in clinical samples, two were obtained from blood cultures, two from respiratory samples, one from urine, one from a burn wound, one from a surgical wound, and two (our strains) from faeces. In almost all cases the patient presented an underlying condition facilitating infection by opportunistic microorganisms. The majority of strains are characterized by their resistance to ampicillin, cephalothin and cefoxitin. Due to the rarity of the isolation of R. aquatilis in human samples it is not yet possible to establish, with any degree of certainty, its true pathogenic capacity.

Child, Preschool↗

Antigen phenotype and cytotoxic activity of natural killer cells in hemodialysis patients.

The antigen phenotype and cytotoxic activity of peripheral blood natural killer (NK) cells have been studied in 35 patients on hemodialysis, evaluating the influence exerted by different types of dialysis membranes and diverse clinical and biologic features of end-stage renal disease. Two-color immunofluorescent analyses with different monoclonal antibodies and flow cytometry were performed to identify the NK cells. Functional cytotoxic assays were simultaneously performed in the same patients. The results show that hemodialysis elicits higher proportions of CD3-/CD56+ and CD3-/CD16+ NK cells and a decrease in their cytotoxic activity compared with the control group. This alteration is related to the use of cuprophan membranes; biocompatible membranes would not significantly affect NK cells. The cuprophan membranes were also found to induce a higher degree of NK cell activation, measured as the number of CD16+/HLA-DR+ cells. Age appeared to be associated with NK cell function. Regarding age, in patients older than 65 years the change from cuprophan to biocompatible membranes was associated with a recovery of NK cytotoxic activity to normal levels. In summary, hemodialysis with cuprophan membranes induces an increased proportion of both NK cells and activated NK cells associated with decreased NK cytolytic activity.

Adult↗

Tumour necrosis factor-alpha (TNF-alpha): the good, the bad and potentially very effective.

When the tumour necrosis factor-alpha (TNF-alpha) gene was cloned the protein became available for use in clinical trials as an antineoplastic agent. However, side effects have severely limited its application in cancer treatment. Studies on the species specificity of TNF have indicated that the p75 TNF receptor (TNFR75) may play an important role in the generation of these side effects in humans. Using human TNF mutants with selective receptor-binding properties it has been demonstrated in neutrophils and endothelium that TNFR75 is involved in the mediation of the proinflammatory activity of TNF by facilitating the p55 TNF receptor (TNFR55). However, only TNFR55 appears to be involved in mediating TNF cytotoxicity. Therefore the potential exists for the successful reintroduction of TNF-alpha, in the form of TNFR55-selective mutants, into the clinical arena with the promise of reduced side effects.

Animals↗

Recombinant desmoglein 3 has the necessary epitopes to adsorb and induce blister-causing antibodies.

The development of an animal model for studying the pathogenesis of pemphigus vulgaris (PV) has been hampered by the unavailability of the purified full-length autoantigen desmoglein 3 (Dsg 3).Therefore, we expressed Dsg 3 using a baculovirus expressed system. The expressed protein was identified as Dgs 3 by its reactivity with a pan-cadherin anti-serum, an anti-serum to a Dsg 3 synthetic peptide, or patient serum, and by amino-terminal sequencing. Carbohydrate analysis showed that recombinant Dsg 3 was glycosylated. While a majority of the recombinant protein was cell associated, by immunoprecipitation, some Dsg 3 was demonstrated in the medium. The Dgs 3 could adsorb out blister-causing antibodies from patient sera. Rabbit anti- Dsg 3 antibodies induced by the recombinant Dsg 3 showed specific binding to intercellular spaces of monkeys esophagus by indirect immunofluorescence. Moreover, these antibodies induced PV-like blisters in neonatal mice and weakly bound perilesional epidermis. Availability of large quantities of relatively pure Dsg 3 should now facilitate studies aimed at understanding Dsg 3 structure and pathogenesis of PV, with implications for developing specific immunotherapies.

Animals↗

Gastroenteritis caused by Aeromonas trota in a child.

A case of acute diarrhoea caused by Aeromonas trota (formerly HG 13 group) in a Spanish child is reported. The strain was isolated in the faeces using the CIN agar (cefsulodin-irgasan-novobiocin) culture media. The strain was initially identified as A sobria by the commercial GNI card and API 20E biochemical systems. The strain was, however, VogesProskauer and sucrose negative, so complementary tests of cellobiose fermentation and gluconate oxidation were performed. These tests, together with the strain susceptibility to ampicillin (MIC 1 microgram/ml) and carbenicillin (MIC < 16 micrograms/ml) led to the final identification of A trota. The microbiological characteristics of this new species and the principal tests required for its identification are presented. The isolation, for the first time, of A trota in the Mediterranean area confirms the suspected worldwide distribution of this species.

Acute Disease↗

Magnetic resonance imaging using FLAIR pulse sequence in white matter diseases.

Fifty six patients among whom 39 had white matter diseases had MRI of the brain comparing FLAIR sequence to a conventional proton density sequence. Flair sequence allowed to detect 18 additional hypersignal (HS) that were not present on T2 sequence. These HS were located in the periventricular areas for 5 of them, near the cortical sulci in 10, and in the centrum semi-ovale for 3. FLAIR sequence permitted analyze 41 other lesions that were not obvious on proton density sequences. Thirty five of them were thus confirmed to be HS : 31 in the paracortical areas, 3 in the paraventricular regions and one in the internal capsule, whereas the remaining 6 were normal sulci of the brain. FLAIR sequence increases the sensitivity of MRI in white matter diseases.

Adolescent↗

Self-association processes involving anthracene labeled phosphatidylcholines in model membrane.

When studying lipid-lipid or lipid-protein interaction in membranes, the correct interpretation of data obtained when using fluorescent phospholipid probes requires the best possible knowledge of probe behaviour in phospholipid membranes. Analysis of the translational dynamics and photochemical properties of the anthracene-labeled phosphatidylcholine (EAPC) shows that a self-association process occurs with this probe in the membrane at the ground state. This anthracene self-association is characterized and leads to a hypochromic effect which has been studied by means of ultraviolet absorption spectroscopy in unilamellar egg-yolk phosphatidylcholine (EggPC) vesicles. A model with indefinite linear self-association, in which each step has the same equilibrium constant, best describes the data. The equilibrium constant was found to be in the 300-500 M(-1) range and the complex lateral distribution pattern of EAPC in model membranes, which results from this self-association process, is characterized and seems to be mainly controlled by the amount of EAPC incorporated into the lipid bilayer.

Journal Article↗

Evisceration after laparoscopic sterilization.

A 35-year-old multiparous woman underwent laparoscopic tubal sterilization without intraoperative problems. Postoperatively, she had nausea and vomiting that failed to respond to antiemetics. The cannula incision site was soaked with serosanguineous discharge. Exploration of the wound revealed evisceration of 10 cm of necrotic small bowel. Resection anastomosis was performed and the patient recovered uneventfully.

Adult↗