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Biomedical subjects

V Alvarez

Publications and source records attributed to V Alvarez.

At least 109 records · Page 6Linked to original sources

[Transferability of results in hematologic determinations].

PURPOSE: To evaluate inaccuracy of the results attained in several Servei Catalá de la Salut laboratories in order to assess if quality purposes are accomplished and results are transferable between them. MATERIAL AND METHODS: Two types of haematological analysers are used: those based upon volumetric measurements and those using light dispersion for particle counting. Inaccuracy was ascertained in accordance to the inter-laboratory quality control programme. The inter-series study was performed with commercial control material in each laboratory with their own data, the monthly coefficient of variation being found, and the mean of the CV of 12 months was then calculated. Intra-series inaccuracy with commercial control material was assessed in each laboratory three times a day for 10 days, and the inaccuracy with fresh patient blood was examined daily in triplicate. The analytical inaccuracy attained in patient samples was also compared with that attained with stabilized control materials. RESULTS: The inaccuracy quality limits were exceeded in the haematological constituents under study. Quality objectives were accomplished in the inter-series inaccuracy study for red cell count, white cell count, MCV, granulocytes and lymphocytes. Repeat fresh patient-blood assays can be used to assess intra-series inaccuracy for red and white cell counts, but not for platelet count. CONCLUSIONS: Inaccuracy of the blood constituents studied is not transferable. Inaccuracy for the commonest haematological values is transferable between laboratories. Intra-series inaccuracy for red and white cell counts can be assessed by repeated use of fresh patient-blood when control material is not available. Patient samples are not advisable as a control of the white cell differential count for intra-series inaccuracy studies in those systems using volumetric principles.

Blood Cell Count↗

A physical map of the human complement component C6, C7, and C9 genes.

The genes for human complement components C6, C7, and C9 are linked on chromosome 5. In this report we describe the physical linkage between C6 and C7 genes. DNA fragments obtained by digestion with several rare-cutting restriction enzymes were separated through pulsed field gel electrophoresis. Hybridization with probes corresponding to the 5' and 3' ends of the three cDNAs showed common bands for the C6 and C7 genes. Both genes are contained in a NotI fragment of 500 kilobases (kb). Moreover, the presence of common 3' C6 and 3' C7 fragments indicates that both genes are oriented in a tail-to-tail, reverse way relative to transcription. No evidence of physical linkage between C9 and C6 or C7 was found in the range 50 kb-2.5 megabases (Mb).

Chromosome Mapping↗

Expression of adhesion molecules in allograft renal dysfunction. A distinct diagnostic pattern in rejection and cyclosporine nephrotoxicity.

We have studied in thirty renal biopsies (from 30 cadaver allograft patients) the expression of both LFA-1 and VLA-4 leukocyte adhesion receptors and their respective ICAM-1 and VCAM-1 endothelial cell ligands, during early allograft dysfunction (24 +/- 5 days after transplantation), reversed either by antirejection therapy (n = 14) or by reduction in CsA dose (n = 16). We have found that the levels of expression of the integrin VLA-4 and the activation signal AIM/CD69 (activation inducer molecule) on interstitial cells were significantly (P < 0.001) higher in rejection than in nephrotoxicity. A main differential expression pattern was observed for VCAM-1, the endothelial cell ligand of VLA-4. Interestingly, a strong staining pattern of the renal vascular endothelium and 35% of tubular epithelium was obtained with anti-VCAM-1 antibody in rejection, as compared with a weak reactivity in endothelium and discrete staining pattern on tubules in nephrotoxicity. On the other hand, we found that the mean percentage of infiltrating cells bearing LFA-1 molecules and the intensity of ICAM-1 (a LFA-1 ligand) expression on endothelium were closely similar in both rejection and CsA nephrotoxicity. Nevertheless, a discrete significant (P < 0.05) "de novo" expression of ICAM-1 was present on tubular cells during rejection. Our results strongly suggest that in rejection the interstitial cell infiltrate seems to be facilitated by the contribution of both LFA-1/ICAM-1 and VLA-4/VCAM-1 cell adhesion mechanisms, and also that VLA-4/VCAM-1 leukocyte interaction does not play a role in cases with CsA nephrotoxicity. Furthermore, the differential expression patterns of VLA-4 and VCAM-1 molecules found between rejection and CsA nephrotoxicity could provide valuable immunohistochemical criteria in the diagnosis of allograft dysfunction.

Adult↗

Effect of endothelin-1 on neutrophil adhesion to endothelial cells and perfused heart.

BACKGROUND: Based on recent evidence showing that endothelin-1 stimulates several activation mechanisms on neutrophils, the aim of the present study was to analyze the effects of endothelin-1 on neutrophil adhesion to endothelial cells and neutrophil accumulation in the heart. METHODS AND RESULTS: The experiments included (1) adhesion of 51Cr-labeled human neutrophils to bovine endothelial cells in culture both in the presence and absence of monoclonal antibodies against the alpha- and beta-subunits of integrins; (2) surface expression of the alpha- and beta-integrin antigens; (3) accumulation of 51Cr-labeled neutrophils on the isolated perfused rabbit heart; (4) in vivo accumulation of autologous neutrophils in the heart, as assessed by myeloperoxidase activity. Endothelin-1 stimulated neutrophil adhesion to endothelial cells (increase of 1 x 10(5) +/- 1 x 10(4) neutrophils per well). The endothelin-1-induced adhesion was blocked (83 +/- 6%) by the anti-CD18 antibody TS1/18 and by several anti-alpha-subunit antibodies. The expression of CD18 and CD11b on the neutrophil surface was also increased by endothelin-1. Endothelin-1 enhanced neutrophil accumulation in the isolated rabbit heart by 4.2 times throughout a TS1/18-inhibitable mechanism. Myeloperoxidase activity increased by 4.2 times in hearts infused in vivo with endothelin-1. CONCLUSIONS: Endothelin-1 stimulates neutrophil adhesion to endothelial cells by an effect on the expression of adhesive molecules on the neutrophil surface. Endothelin-1 stimulates neutrophil accumulation in vivo and in vitro in the heart. Antibodies against the integrin complex block the endothelin-1-dependent neutrophil adhesion. These findings have potential importance in the pathophysiology of endothelin-1-increased states.

Animals↗

Altered blood pressure during sleep in normotensive subjects with type I diabetes.

This study was designed to examine the circadian pattern of blood pressure in children and young adults with type I diabetes who were completely normotensive by standard criteria. Forty-five patients and the same number of age- and sex-matched control subjects were studied. In diabetic children of 10-14 years of age, the nocturnal fall in systolic and diastolic blood pressures was intact. In diabetics of 15-20 years of age, the fall in systolic blood pressure was blunted; in diabetics of 21-37 years of age, the fall in both systolic and diastolic blood pressures during sleep was blunted. When data from all diabetic subjects were pooled and analyzed in a multiple linear regression model, mean blood pressure during sleep correlated best with urinary albumin excretion (r = 0.60). On the basis of this finding, we subdivided our patients into two groups: a microalbuminuric group (urinary albumin excretion > 30 mg per 24 hours; mean, 160.3 +/- 29.7; n = 11) and a normoalbuminuric group (urinary albumin excretion < 30 mg per 24 hours; mean, 6.6 +/- 6.5; n = 34). Both systolic and diastolic blood pressures during sleep were higher in microalbuminuric (121.1 +/- 3.3 and 69.3 +/- 2.5 mm Hg, respectively) than in normoalbuminuric diabetics (114.2 +/- 1.8 and 60.1 +/- 1.2 mm Hg, p < 0.05) or control subjects (113.3 +/- 1.2 and 60.1 +/- 1.2 mm Hg, p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Approaches for providing target values to improve usefulness of external quality assessment scheme. The Spanish experience.

The aim of this communication is to highlight the specific aspects of external quality assessment schemes that need to be discussed in a European context: target values, transferability of results and accredit of laboratories. The Spanish situation is presented here. The most reliable way to provide target values is to analyse the control samples by reference methods. However, it is not possible for the majority of national schemes and other approaches are presently used: the verification of consensus means is a practicable solution adopted in Spain. An initial network involving selected routine laboratories has been developed, to attain transferability of results. The traceability of routine calibrators from certified reference materials should be demonstrated. To accredit laboratories for licensing is a complex activity that should consider many aspects, results from the national quality assessment scheme bring one. A scoring system is being used in Spain for guidance, and the complete guidelines are under preparation.

Accreditation↗

Biochemical and functional characterization of the leucocyte tyrosine phosphatase CD45 (CD45RO, 180 kD) from human neutrophils. In vivo upregulation of CD45RO plasma membrane expression on patients undergoing haemodialysis.

The biochemical and functional characterization, and the regulation of plasma membrane expression of the leucocyte tyrosine phosphatase CD45, have been investigated in neutrophils from healthy donors and patients undergoing haemodialysis. CD45 proteins of 180 kD and 130-150 kD were precipitated from neutrophils from both healthy subjects and haemodialysed patients. Prolonged storing, as well as trypsin treatment of samples containing the 180-kD CD45 protein, generated the 130-150-kD polypeptides. The 130-150-kD CD45 polypeptides carried extracellular CD45 epitopes, including the sialic acid-related UCHL1 epitope (CD45RO). Furthermore, these trypsin-generated CD45 polypeptides did not possess phosphatase activity, which could be detected on the 180-kD protein. A remarkable quantitative increase of cell surface expression of the neutrophil CD45 components was detected both after in vitro neutrophil activation and after dialysis treatment with neutropenic membranes. The CD45 biochemical pattern did not qualitatively change upon either in vitro or in vivo dialysis-induced neutrophil activation. The upregulated expression of CD45 on neutrophils from dialysed patients correlated with the neutropenic effect induced by the different dialyser membranes. Maximal upregulation of CD45 expression was observed after 15 min of dialysis with neutropenic membranes, and normal expression levels were restored after 1 h. By contrast, increase of CD45 plasma membrane expression induced in vitro by treatment of normal neutrophils with the degranulatory agents fMLP or Ca2+ ionophore was maintained. These results demonstrate that neutrophil cell surface expression of the 180-kD CD45 protein is upregulated during the in vivo haemodialysis process, and suggest that a proteolytic activity could regulate the enzymatic activity of CD45 by degranulation of its cytoplasmic phosphatase domains.

Antigens, CD↗

Development of systemic lupus erythematosus in a patient on hemodialysis.

A case of systemic lupus erythematosus (SLE) that developed 2 years after beginning hemodialysis is reported. The patient had not been given any drug implicated in the production of SLE. She had been treated with deferoxamine, an in vitro inhibitory of DNA synthesis. The difficulty of the diagnosis is emphasized. Clinical improvement after prednisone treatment was impressive. SLE may appear even in patients receiving hemodialysis, despite immunological depression derived from chronic uremia.

Adolescent↗

Long-term neuromuscular dysfunction produced by passive transfer of amyotrophic lateral sclerosis immunoglobulins.

We investigated the role of the immune system in the pathogenesis of amyotrophic lateral sclerosis (ALS) by studying the long-term consequences of ALS immunoglobulin (Ig) application on the levator auris muscle of the mouse. We applied Ig from seven ALS patients, four disease controls, and a pool of normal Ig (6 mg of Ig in 2 weeks) by subcutaneous injection; removed the muscles 4 to 12 weeks after the beginning of treatment; and recorded both spontaneous and evoked release of transmitter. None of the control Ig induced changes in transmitter, whereas five of seven ALS Ig induced a significant increase in the rate of spontaneous release, and all ALS Ig produced significant changes in the quantal content of evoked release. In muscles treated with one of the ALS Igs, synaptic activity was completely absent. Cholinesterase and silver staining demonstrated intact neuromuscular junctions in the control Ig-treated muscles and also in many areas of ALS Ig-treated muscles. Axonal degeneration and denervation were present in most muscles treated with ALS Ig. There was complete denervation when no synaptic activity could be recorded. Thus, ALS Ig appears to lead to long-lasting effects at the neuromuscular junction, and such effects may be an early stage in the immune-mediated pathogenesis of ALS.

Adult↗

Differentially regulated cell surface expression of leukocyte adhesion receptors on neutrophils.

To elucidate the molecular mechanisms accounting for hemodialysis-induced neutropenia, the regulation of plasma membrane expression of leukocyte adhesion glycoproteins was investigated by both flow cytometry and immunoprecipitation techniques. The members of the LFA family of integrins, Mac-1/Mo1 (CD11/CD18) and gp150/95 (CD11c/CD18), involved in adhesion of myeloid cells to endothelia and other substrates, were found to be overexpressed on the plasma membrane of neutrophils from patients undergoing hemodialysis with a Cuprophane dialyzer, whereas no change was observed in the expression of LFA-1 (CD11a/CD18). By contrast, dialysis with Cuprophane membranes, as well as in vitro treatment with different activating agents, induced a downregulation on the expression of both the Leu-8/LAM-1 antigen, the human neutrophil peripheral lymph node homing receptor, and the CD43 major sialoglycoprotein involved in leukocyte homotypic adhesion. Kinetics studies showed that these up- and downregulatory processes of antigen expression occur very rapidly, correlating with maximal neutropenia. Recovery of initial levels of expression of CD11b/CD18 and Leu-8/LAM-1 adhesion molecules was observed after one hour of hemodialysis. However, the basal expression of CD43 was not restored by that time. The coordinated upregulation of CD11b and CD11c and downregulation of LAM-1 and CD43 adhesion receptors provide molecular mechanisms for understanding leukoaggregation, adherence to endothelia, and extravasation of neutrophils ultimately leading to the hemodialysis-induced neutropenia.

Cell Membrane↗

Peak expiratory flow measured with the Mini Wright Peak Flow Meter in children.

We established pediatric reference values for peak expiratory flow rate (PEFR), using the Mini Wright Peak Flow Meter. The study was based on 1,566 Mediterranean white children, aged 7 to 14 years from Valencia (Spain) schools. Height was the biometric variable with the greatest correlation to PEFR in both sexes; significant differences were noted between males and females. Prediction equations and percentile tables are presented for each sex. The performance of the Mini Wright Peak Flow Meter was compared with that of a spirometer.

Adolescent↗

Serological markers and celiac disease: a new diagnostic approach?

We analyze the diagnostic efficacy of two celiac disease serological markers: anti-gliadin (IgA and IgG class) and anti-endomysium IgA-class (EmA-IgA) antibodies applied to 336 serum samples from celiac patients on both gluten-challenge and gluten-free diets, and a control group. The anti-gliadin and anti-endomysium antibodies levels were significantly higher among the gluten-consuming celiac patients than in the other groups. The greatest efficacy in diagnosing celiac disease was achieved in the Ema-IgA class test. The IgA-class anti-gliadin antibodies proved to be more specific, with a higher positive test predictive value than the IgG-class anti-gliadin antibodies, whereas the latter proved to be more sensitive, with a higher negative test predictive value than those of the IgA-class anti-gliadin antibodies. Our results also demonstrate that the simultaneous assessment of anti-gliadin IgA- and IgG-class antibodies constitutes a valid test in selecting patients suspected of having celiac disease. In turn the EmA-IgA antibodies constitute a confirmative test for indication of an intestinal biopsy.

Adolescent↗

A new case of partial trisomy 3(q25----qter) in a newborn.

A description of a female newborn with some physical stigmata presenting a partial trisomy for 3(q25----qter) is given. The additional material is located at the distal end of the short arm of chromosome 15 as the result of a maternal t(3;15) (q25:p13). Some characteristics of the proband were similar to those described for the 3(q21----qter) trisomy.

Abnormalities, Multiple↗

Radioimmunoscintigraphy and radioimmunotherapy of renal cell carcinoma xenografts.

From a panel of monoclonal antibodies (MAb) reactive with human renal cell carcinoma generated by this laboratory, three (designated A6H, C5H, and D5D) were selected for in vivo studies with a nude mouse xenograft model. These studies included 131I- and 111In-labeled MAb radioimmunoscintigraphy and 131I-labeled MAb radioimmunotherapy. In the imaging studies, these radiolabeled MAb allowed visualization of subcutaneous xenografts larger than 40 mg and subrenal capsule xenografts smaller than 20 mg. Comparisons of tumor to non-tumor tissue radiolabel distribution yielded unusually high ratios and depended on the MAb-xenograft combination. The 111In-radiolabeled A6H showed increased accumulation in the liver compared with 131I-A6H, but this still did not necessitate background subtraction for good visualization of small, subrenal capsule renal cell carcinoma xenografts. Radioimmunotherapy studies with 131I-A6H in BALB/c nu/nu mice bearing established renal cell carcinoma xenografts showed a prolonged (greater than 90 days) regression in tumor burden and possible "cures," whereas three sets of control mice showed progressive and rapid increases in tumor size. These studies indicated that MAb, which show good tissue biodistribution and high imaging sensitivity, could also be capable of delivering effective radiotherapy to the tumor when "human equivalent" radiolabeled-MAb doses are used.

Animals↗