Search PubMed⌕ Search

Biomedical subjects

M Contreras

Publications and source records attributed to M Contreras.

At least 19 recordsLinked to original sources
PubMed1998

Intracellular cytokine profile of cord and adult blood lymphocytes.

Umbilical cord blood (CB) transplantation is thought to be associated with a reduced risk of severe graft-versus-host-disease (GVHD) compared with bone marrow transplantation (BMT). The cytokine cascade is known to be important in the pathogenesis of GVHD; however, previous studies investigating the cytokine secretion pattern of CB cells have been contradictory because of variations in experimental techniques. In this study, the cytokine profile of cord and adult blood lymphocytes and lymphocyte subsets has been assessed at the single-cell level by flow cytometry, using CD4/CD8 and CD45RA/CD45RO markers. Cord and adult blood mononuclear cells were stimulated with phorbol 12-myristate 13-acetate (PMA) and ionomycin in the presence of monensin. After 4 to 24 hours of incubation, interleukin-2 (IL-2), IL-4, interferon-gamma (IFN-gamma), and tumor necrosis factor-alpha (TNF-alpha) production was measured by three-color flow cytometry. The results show that cord blood lymphocytes (CBL) produce less IL-2, IL-4, IFN-gamma, and TNF-alpha than adult peripheral blood lymphocytes (ABL). Further subset analysis showed that in CBL the majority of cytokine producing cells were CD4(+)CD45RA+, whereas in ABL the cytokine-producing cells were both CD4(+)CD45RO+ and CD8(+)CD45RO+. These results suggest that the reduced incidence of GVHD in CB transplantation may partly due to the altered cytokine profile seen in CBL.

Adult↗
PubMed1998

The London Cord Blood Bank.

Cord blood (CB) contains a sufficient number of haemopoietic progenitor cells and can be used as an alternative to bone marrow transplantation. To date, a number of transplants using cord blood-derived stem cells from HLA-identical siblings and unrelated donors have been successfully performed. However, only approximately 30% of patients have an HLA-identical sibling while the rest are reliant on finding an HLA-compatible unrelated donor. This is particularly difficult for patients from ethnic minorities since the majority of unrelated bone marrow donor volunteers are from Caucasoid ancestry. The London Cord Blood Bank was established to provide an additional source of stem cells for all patients requiring bone marrow transplantation.

Blood Banks↗
PubMed1997

The impact of a picture archiving and communication system on nuclear medicine examination interpretation.

Radiographic correlation is essential for many of the examinations performed in nuclear medicine. The purpose of this study was to evaluate the impact of a picture archiving and communications system (PACS) on the function and efficiency of a nuclear medicine department at a tertiary care institution. We evaluated 250 consecutive noncardiac nuclear medicine imaging examinations and asked the interpreting physician the following questions: (1) Was PACS used in the interpretation of the study? (2) Did the use of PACS expedite examination completion or aid in study interpretation? And (3) Did the use of PACS permit a definitive diagnosis to be made? PACS was accessed for correlative radiographic images in 155 of the 250 (62%) nuclear medicine examinations. Images available on PACS for review aided in study interpretation in 74% (115 of 155) of cases. The use of PACS was thought to expedite examination completion in 55% (86 of 155) of cases. The system was accessed but not operational in only 1% of cases (2 of 155). PACS provides reliable, rapid access to multimodality correlative radiographic images that aid in the interpretation of nuclear medicine examinations. Such systems also increase the efficiency of a nuclear medicine service by allowing timely and conclusive interpretations to be made.

Evaluation Studies as Topic↗
PubMed1997

Comparison of in vivo red cell survival of donations collected by Haemonetics MCS versus conventional collection.

The Haemonetics Multicomponents System (MCS) cell separator allows concurrent donation of red cells in addition to platelets and/or plasma, thus increasing the versatility of apheresis donations. In vivo survival of autologous red cells obtained by MCS was compared with red cells collected conventionally. In this cross-over controlled study, five male volunteers donated one unit of red cells by MCS and one unit of whole blood by the conventional manual method, 3 months apart. After storing donations in SAG-M for 35 days under standard conditions, radioactive (51Cr)-labelled autologous red cells were injected into each donor. The post-transfusion recovery (PTR) of red cells at 24 and 48 h did not show any significant difference between red cells obtained manually and by MCS, indicating that processing differences have no detrimental effects on red cell survival.

Blood Donors↗
PubMed1996

Topology of ATP-binding domain of adrenoleukodystrophy gene product in peroxisomes.

Adrenoleukodystrophy (X-ALD) is a demyelinating disorder characterized by the accumulation of saturated very-long-chain fatty acids (> C22:0) due to the impaired activity of lignoceroyl-CoA ligase. The gene responsible for the disease was found to code for a 84-kDa peroxisomal integral membrane protein. Its amino acid sequence has high homology with the ATP-binding cassette superfamily of transporters and it is predicted to have six membrane-spanning segments and a putative ATP-binding domain. To define the function of ALDP, we studied the topology of its ATP-binding domain by using antibodies (1D6) against a hydrophobic domain (amino acid residues 279 to 482) and antibodies (Abct) against the C-terminal 15-amino-acid hydrophilic domain (amino acid residues 731 to 745) of ALDP. The observation of punctate fluorescence in permeabilized ALD fibroblasts, using Abct antibodies but not with antibodies against catalase, suggests that the C-terminal segment of ALDP is projected toward the cytoplasm from the peroxisomal membrane. Trypsinization of intact peroxisomes under isotonic conditions abolishes the Abct antibody recognition site, whereas the 1D6 antibodies identify a degradation product of 43-kDa protein that has been protected and retained by the membrane. This again suggests that the C-terminal portion of the ALDP protein is located on the outside (cytoplasmic) face of the peroxisomal membrane. Additional support for this conclusion was obtained by purification of the ALDP C-terminal domain, released from purified rat liver peroxisomes incubated with the cytosolic fraction, using blue-Sepharose affinity chromatography. A 47-kDa peptide retained by the column was recognized by Western blot analysis with Abct antibodies against the C-terminal sequence of ALDP and this polypeptide on polyvinylidene difluoride membrane was able to bind [gamma-32P]ATP in vitro in the presence of Mg2+. These results demonstrate that the C-terminal peptide containing the ATP-binding domains of ALDP is on the cytoplasmic surface of the peroxisomal membrane where this domain may function as an ATPase to support the functional role of ALDP in the peroxisomal membrane.

ATP Binding Cassette Transporter, Subfamily D, Mem↗
PubMed1996

Determination of imidacloprid in vegetables by high-performance liquid chromatography with diode-array detection.

An HPLC method is described for the determination of imidacloprid residues in vegetables at levels ranging from 0.01 to 0.60 mg/kg. The selection of the extraction and clean-up procedure is discussed. Spectral data obtained with diode-array detection allow the identification of imidacloprid residues. Thermospray mass spectrometric studies were carried out in combination with HPLC. The mean recoveries and standard deviations were 95% and 4.7%, respectively, in the various crops tested. Registration of the analytical results for a control sample in quality control charts demonstrated the performance of the method. Data for incurred residues of imidacloprid in vegetable samples routinely applying the proposed method are also presented.

Chromatography, High Pressure Liquid↗
PubMed1996

Evaluation of anastomotic hyperplasia progression using the cyclin specific antibody MIB-1.

BACKGROUND: Delayed failure of prosthetic arterial grafts is primarily due to the development of anastomotic intimal hyperplasia. This report follows the proliferation of smooth muscle cells that ensues after prosthetic arterial grafting, using the cyclin-specific antibody MIB-1. METHODS: Six-mm expandable polytetrafluoroethylene (ePTFE) grafts were placed end-to-end in the carotid arteries of mongrel dogs. Animals were randomly assigned to sacrifice intervals of 2, 7, 14, and 30 days. Serial coronal sections were cut and immunohistocytochemistry performed using the MIB-1 antibody. RESULTS: The control carotid artery had no definable proliferation. Two days after grafting, there was brisk proliferation in the upper one third of the arterial media. By 7 days, proliferation and migration of smooth muscle cells was seen above the internal elastic lamina, in which 50% of the cells were MIB-1 positive. Fourteen days after graft placement, proliferation continued in the neointima; however, the proliferation index was diminished compared with previous time intervals. At 30 days, despite a dramatic increase in lesional increase, there was a marked decrease in the overall proliferation of cells. CONCLUSIONS: Following placement of a prosthetic arterial graft, there is initial brisk proliferation of cells in the arterial media, with migration, ongoing proliferation, and resultant development of a localized cellular neointima. Over a 30-day period, the percentage of cells proliferating subsides in contrast to the progressive increase in the size of the neointima. Immunohistocytochemistry with the MIB-1 antibody is a useful tool in defining the cellular kinetics after prosthetic arterial grafting.

Anastomosis, Surgical↗
PubMed1996

Cytokine levels in platelet concentrates: quantitation by bioassays and immunoassays.

Some adverse reactions to the transfusion of platelet concentrates (PCs) cannot be attributed to antibodies against blood cells or to subclinical microbial agents. It has been suggested that leucocyte-derived inflammatory cytokines such as interleukin (IL)-1, IL-6 and tumour necrosis factor (TNF) may contribute to a larger number of unexplained non-antibody-mediated adverse reactions. Three types of PCs, containing different levels of leucocytes, are currently produced. Filtration is used on demand to further reduce leucocyte contamination of these components. we have monitored the plasma of PCs prepared by the platelet-rich plasma method (PRP), the buffy-coat method or by apheresis for IL-6, IL-1, transforming growth factor-beta (TGF-beta), TNF and interferon gamma (IFN gamma). Biologically active IL-6 increased in stored PRP-PCs from a mean of 140 pg/ml on day 1 to 2395 pg/ml on day 5/6. Elevated levels of IL-8, as detected by immunoassay, were evident in PRP-PCs during routine storage under blood bank conditions. Small amounts of immunoreactive IL-1 with only minimal biological activity were present in some PRP-PCs by day 5/6. No significant increase in the levels of IL-8, IL-6 or IL-1 were seen in buffy-coat PCs during storage for 5/6 d. For apheresis PCs, an increase in IL-8 content, but not in IL-6 over 6 d was observed. In all three types of PCs, elevated amounts of both bioactive and immunoreactive TGF beta were present, but there was no evidence of any biologically active or immunoreactive TNF alpha. Pre-storage filtration of PRP-PCs for depletion of leucocytes prevented the increase in IL-8 and IL-6 levels of these PCs. Our results show that leucocyte reduction by buffy-coat method reduces cytokine levels to a comparable level to filtered or apheresis PCs, containing low levels of leucocytes, but use of these PCs in minimizing the severity and incidence of reactions in recipients will require clinical evaluation. This is the first comprehensive and comparative study which, on the basis of biological activity of cytokines, directly indicates that the mode of platelet production grossly influences the levels of cytokines.

Biological Assay↗
PubMed1996

Myelodysplastic syndrome with karyotype abnormality is associated with elevated F-cell production.

A sensitive F-cell assay has been used to examine the production of fetal haemoglobin (Hb F) in a group of 77 adult patients with myelodysplastic syndrome (MDS), and a control group composed of 100 normal blood donors. Although the mean F-cell percentage in the MDS group (6.0%) is not statistically different from that in the normal blood donors (3.1%), a higher proportion of myelodysplastic patients have elevated F-cell values and the magnitude of the increases is greater than that observed in blood donors. In order to investigate the association further, the karyotypes of the MDS patients have been examined. 13/21 (61.9%) of the MDS patients with karyotypic abnormalities have F-cell values > 5%, compared to only 6/56 (10.9%) of the MDS patients with a normal karyotype and 11/100 (11%) of the blood donors. The observed difference in the distributions of F cells between the two subgroups of patients with MDS is highly significant (P < 0.0001).

Adult↗
PubMed1996

A prospective study of the incidence of red cell allo-immunisation following transfusion.

To establish the incidence and timing of red cell allo-immunisation following transfusion, pretransfusion and serial post-transfusion samples were screened for allo-antibodies in a total of 452 patients who had undergone elective surgery. Antibody screening was performed by 2-stage papain, manual polybrene and indirect antiglobulin techniques (IAT). Red cell allo-antibodies were found in 42 patients and 38 of these (8.4% overall) demonstrated antibodies only after transfusion; 76% of them had Rh specificity. This rate of red cell allo-immunisation is higher than what would be expected if transfused patients were tested only once post-transfusion, as has been the case in several previous studies. For the type of patients studied, this finding may not be of clinical relevance at present because most patients undergoing elective surgery do not require further transfusion in their lifetime. However, this is changing with the longer life expectancy of the population and the increased probability of repeat surgery. Twenty-two (58%) of the antibodies were initially detected by the 2-stage papain and/or polybrene techniques, when the IAT was negative, although later 19 became positive by IAT; this added sensitivity of techniques other than the IAT, to detect early allo-immunisation may be relevant in pretransfusion testing to prevent haemolytic transfusion reactions in patients requiring repeated transfusions.

Adult↗
PubMed1996

Bronchoalveolar oxyradical inflammatory elements herald bronchopulmonary dysplasia.

OBJECTIVES: To quantify oxyradical inflammatory markers in serial endotracheal tube aspirates obtained from premature neonates at risk for developing bronchopulmonary dysplasia, and to correlate these parameters with clinical manifestations of the disease. DESIGN: Prospective cohort study. SETTING: Tertiary neonatal intensive care unit. PATIENTS: Twenty-eight intubated, premature infants, with 15 infants displaying simple respiratory distress syndrome and 13 infants eventually developing bronchopulmonary dysplasia. INTERVENTIONS: Endotracheal tube aspirates were collected and clinical severity scores were calculated longitudinally from an inception cohort during the first week of life. Diagnosis of bronchopulmonary dysplasia by standard criteria was recorded at 30 days of life. Various biochemical analyses related to pulmonary oxyradical stress were determined on endotracheal tube aspirates and were normalized according to the magnitude of serum/aspirate urea ratios. The demographic, illness severity, and biochemical characteristics of infants with simple respiratory distress syndrome and those characteristics of infants developing bronchopulmonary dysplasia were evaluated by masked comparison. MEASUREMENTS AND MAIN RESULTS: Populations of respiratory distress syndrome and bronchopulmonary dysplasia infants could be differentiated during the first week of life by means of the following parameters: gestational age; birth weight; Score of Neonatal Acute Physiology; Neonatal Therapeutic Intervention Scoring System; epithelial lining fluid leukocytes; elastase; myeloperoxidase; xanthine oxidase and catalase enzyme activities; and total sulfhydryls. CONCLUSIONS: Infants with simple respiratory distress syndrome could be segregated from those infants who developed bronchopulmonary dysplasia by the magnitude of the epithelial lining fluid oxyradical inflammation markers. While infants developing bronchopulmonary dysplasia typically exhibited increased concentrations of these markers during the first week of life, those infants with simple respiratory distress syndrome displayed low, uniform, or decreasing values of these markers over this interval. Infants developing bronchopulmonary dysplasia demonstrate an early pulmonary inflammatory response, and one key aspect of this response involves various oxyradical-generating systems.

Bronchi↗