No evidence for protein modifications in fresh frozen plasma after photochemical treatment: an analysis by high-resolution two-dimensional electrophoresis.

To study if photochemical treatment of human plasma by methylene blue in combination with visible light induces protein alterations, we analysed by high-resolution two-dimensional gel electrophoresis (2-D PAGE) untreated and photochemically treated fresh frozen plasma collected by apheresis from the same donor (n = 8). Polypeptides were separated according to their charges by isoelectric focusing and to their sizes in polyacrylamide gels in presence of sodium and to their sizes in polyacrylamide gels in presence of sodium dodecyl sulphate, and visualized by silver staining. Despite their apparent complexity, protein maps of untreated plasma and photochemically treated fresh frozen plasma revealed identical spot patterns.

[Antiphospholipid and anti beta 2-glycoprotein I antibodies].

Recent studies have shown that beta 2glycoprotein I (beta 2GPI), a plasma inhibitor of coagulation with affinity for anionic phospholipids, is frequently required for the formation of the epitopes recognized by some anti-phospholipid antibodies. Six murine monoclonal antibodies directed against human beta 2GPI were compared with anti-beta 2GPI antibodies associated with systemic lupus erythematosus and primary anti-phospholipid syndrome. The beta 2GPI-dependent binding properties to neutral and anionic phospholipids were studied, as well as the target epitopes on the beta 2GPI molecule and the effect of various beta 2GPI treatments to get rid of contaminating phospholipids or to block the beta 2GPI-phospholipid interaction.

Two-dimensional electrophoresis as an aid in the analysis of the clonality of immunoglobulins.

In this paper, we summarize our five-year observation of two-dimensional polyacrylamide gel electrophoretic (2-D PAGE) analysis of immunoglobulin (Ig) light (L) chain patterns on serum/plasma and/or purified human Ig, and compare this technique with agarose electrophoresis and/or immunofixation examination. Polyclonal Ig L chains were seen as large "fuzzy" areas with several zones of high density. The majority (71%) of the monoclonal Ig L chains of monoclonal gammopathy detected by conventional electrophoresis appeared as a single large and well-defined spot on 2-D PAGE analysis, with the remaining appearing as multiple spots.

Peripheral blood stem cells harvested after chemotherapy and GM-CSF for treatment intensification in patients with advanced lymphoproliferative diseases.

Peripheral blood stem cells (PBSC) from 15 patients with advanced non-Hodgkin's lymphoma (NHL), two patients with chronic lymphocytic leukemia, and two patients with myeloma were collected by continuous-flow leukapheresis after chemotherapy with MIV (mitoxantrone, ifosfamide, and etoposide, five patients) or high-dose cyclophosphamide (14 patients), followed by administration of GM-CSF. Sixteen patients (84%) had persistent marrow involvement at time of inclusion. Results were compared to those obtained in a control group of similar age and disease status in whom collection had been performed after MIV chemotherapy alone.

Circadian rhythms of circulating NK cells in healthy and human immunodeficiency virus-infected men.

Antiviral immunity involves NK cells, which circulate rhythmically every 24 hours. We have investigated circadian and 12-hour rhythms in the peripheral count of circulating NK cells in 15 men infected with human immunodeficiency virus (HIV) and 13 healthy controls. We analyzed three phenotypes using double-labeling with monoclonal antibodies and flow cytometry assessment: CD3- CD16+, CD3-CD57+, and CD2+CD3-.

Dermatan sulfate inhibition of fibrin-rich thrombus formation in nonhuman primates.

Dermatan sulfate (DS), a factor that amplifies plasma heparin cofactor II antithrombin (HCII) activity, has been evaluated in baboons for its relative antithrombotic and antihemostatic effects by use of a model that combines both platelet-rich and fibrin-rich thrombus formation. Thrombus was generated in a two-component thrombogenic device incorporated into exteriorized femoral arteriovenous shunts, in which a proximal segment of collagen-coated tubing induces platelet-rich arterial-type thrombus and distal expanded chambers with disturbed and static flow produce fibrin-rich venous-type thrombus. Thrombus formation was measured as the deposition of autologous 111In-platelets by imaging analysis and by the accumulation of 125I-fibrin.

The spectrum of beta-thalassaemia in Algeria: possible origins of the molecular heterogeneity and a tentative diagnostic strategy.

We report here on the final results of an epidemiological survey involving 177 beta-thalassaemic chromosomes in Algeria. Four common mutations account for 86% of the chromosomes, the other ones carrying nine other rare mutations. Combination of these results with those of other smaller regional epidemiological studies indicates the existence of still a wider range of mutations.

High expression of bcl-2 protein in acute myeloid leukemia cells is associated with poor response to chemotherapy.

The BCL-2 proto-oncogene encodes a mitochondrial protein that blocks programmed cell death. High amounts of bcl-2 protein are found not only in lymphoid malignancies, but also in normal tissues characterized by apoptotic cell death, including bone marrow. Using a monoclonal antibody to bcl-2 protein, we analyzed 82 samples of newly diagnosed acute myeloid leukemia.

Evaluation of six markers of haemostatic system in normal pregnancy and pregnancy complicated by hypertension or pre-eclampsia.

Objective: To establish the plasma evolution of prothrombin fragments 1+2 (F 1+2), thrombin-antithrombin III complexes (TAT), fibrin fragment D-Dimers (DD), von Willebrand factor antigen (vWf), Type 1 plasminogen activator inhibitor antigen (PAI) and blood platelet count during normal pregnancy and to compare these values with those obtained in hypertensive or pre-eclamptic pregnancies.

Design: Cross-sectional study.

Subjects: Forty-seven healthy pregnant women with gestational age ranging between 5 and 40 weeks, and fourteen women with gestational age ranging between 25 and 38 weeks presenting with either gestational hypertension (n = 4) or pre-eclampsia (n = 10).

Antithrombotic properties of a dermatan sulfate hexadecasaccharide fractionated by affinity for heparin cofactor II.

The relationship between the antithrombotic activity of dermatan sulfate (DS) in vivo and its catalytic effect on the inhibition of thrombin by heparin cofactor II (HC II) in vitro was investigated. DS was depolymerized by Smith degradation and the fragments obtained were separated by gel filtration. The fragment of minimal size with full catalytic activity was a hexadecasaccharide, which was further fractionated by affinity for immobilized HC II.

[Acquired autoantibodies directed against human factor VIIIc. Treatment of 2 hemorrhagic accidents with porcine factor VIIIc].

The emergence of an autoantibody directed against factor VIII may be responsible for severe, life-threatening haemorrhages. This rare disease is usually idiopathic, but it may be consecutive to an autoimmune disease or to the absorption of certain drugs such as penicillin. The diagnosis rests on the finding of a prolonged activated thromboplastin time with presence of a circulating anticoagulant and deep fall in factor VIII level.

Clonal imbalances of plasma/serum immunoglobulin production in infants.

To study the clonal events occurring during ontogeny of the humoral immune system, we evaluated plasma immunoglobulin (Ig) production in term newborns and young children by high-resolution two-dimensional gel electrophoresis. The clonality pattern of Ig light (L) chains from healthy newborns (n = 19) was similar to that observed on protein maps of their mothers or of normal adults (n > 100), that is, rare distinguishable small spots in a cloud-like large band of indiscrete Ig L chain spots (polyclonal pattern; maternal Igs). Analysis of plasma samples obtained from infants between 1 month and 5 years of age (n = 55) revealed discrete but evident alterations of the clonality of Ig production.

Pattern variations of polyclonal and monoclonal immunoglobulins of different isotypes analyzed by high-resolution two-dimensional electrophoresis.

High-resolution two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) was used to analyze serum samples and purified immunoglobulins (Ig) obtained from "normal" individuals and from patients diagnosed with monoclonal gammopathies (MG) (n = 47; 5 IgA, 15 IgM, 15 IgG, 4 biclonal IgG, 1 IgD, 7 Bence Jones proteins). Polyclonal and monoclonal heavy (H) chains were located at different restricted gel positions according to their isotype. Monoclonal H chains appeared as sets of spots characterized by charge (pI) and size (M(r)) microheterogeneity.

[ALAT and viral RNA as risk factors in 68 blood donors with anti-hepatitis C antibodies].

Aim: Determine the risk factors in blood donors with anti hepatitis C antibodies (anti-HCV ab) possible liver involvement and evaluation of their infectious potential by a search for viral RNA in blood.

Methods: Between July 1990 and October 1991, 19,632 blood donors were screened for hepatitis C. Antibodies to HCV were detected in 74 donors (2nd generation ELISA, Abbott).

Plasma/serum protein patterns in human fetuses and infants: a study by high-resolution two-dimensional polyacrylamide gel electrophoresis.

Plasma/serum proteins of fetal blood samples (N = 88) obtained under ultrasound guidance between the 18th and the 39th week of pregnancy, of blood samples collected from premature infants (N = 19), newborns at term (N = 20) and children of less than 5 years of age (N = 55) were analysed by high-resolution two-dimensional polyacrylamide gel electrophoresis. By comparison with adult 'reference' protein maps, tens of different proteins (and some of their genetic variants) were identified on the electrophoretograms. After the 18th week of gestation, albumin, transferrin, Factor B, glu- and lys-plasminogen, antithrombin III, Gc-globulin, alpha 1-antitrypsin, alpha 2-HS-glycoprotein, several apolipoproteins (apo A-I, A-II, A-IV, C-II, C-III, D, E, J), retinol-binding protein, transthyretin and alpha-fetoprotein could be observed.

Production of stable human-mouse hybridomas secreting monoclonal antibodies against Rh D and c antigens.

Peripheral blood lymphocytes from donors immunized against Rh antigens were fused with mouse myelomas and heteromyelomas in order to obtain human-mouse hybridomas secreting antibodies specific for these antigens. Three cell lines secreting anti-D IgG and two secreting anti-c IgM were stabilized and produced immunoglobulins for several months. These human monoclonal antibodies were evaluated as reagents for Rh phenotyping.

Immunological characteristics of umbilical cord blood cells: phenotypic and functional analysis.

The purpose of this study was to phenotype and assay immunological function on cord blood from 70 samples. Immunophenotyping indicated similar numbers of CD2, 3, and 8-positive cells as in adult bone marrow. CD4-positive cells were increased and CD19, 20, and CD56-positive cells were decreased in numbers.

Effect of prestorage leukocyte reduction on proteins of platelets obtained by apheresis.

The effect of prestorage leukocyte reduction was evaluated on platelet concentrates (PCs) obtained by apheresis using the 'surge' technique. Two hours after collection, the PCs were divided into 2 equal units. One unit was filtered through a Sepacell PL-10a, producing a filtered PC (FPC).

Effects of heparin, dermatan sulfate and of their association on the inhibition of venous thrombosis growth in the rabbit.

This study compares the ability of unfractionated heparin, of dermatan sulfate, and of their simultaneous administration delivered as continuous intravenous infusion or as a single bolus injection to inhibit the growth of a standardized venous thrombosis in the rabbit. When delivered as continuous intravenous infusion for 4 h, heparin and dermatan sulfate inhibited thrombus growth in a dose dependent manner. The maximum antithrombotic effect of heparin was achieved at the dose of 0.

Clonal imbalances of serum immunoglobulins after allogeneic bone marrow transplantation: an analysis by high-resolution two-dimensional gel electrophoresis.

The clonality pattern of immunoglobulins (Igs) produced after allogeneic bone marrow transplantation (BMT) was studied by high-resolution two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) of serum samples and purified Igs. With this technique, the light (L) chain of a monoclonal Ig usually appears as a single spot. Thus, the degree of clonal diversity of the functional B cells can be appreciated by the electrophoretic pattern of the serum L chains.

Black beta-thalassemia homozygotes with specific sequence variations in the 5' hypersensitive site-2 of the locus control region have high levels of fetal hemoglobin.

We have sequenced the 5' hypersensitive-2 (5'HS-2) site of the locus control region (LCR) and the promoters of the two gamma-globin genes located on chromosome 11 of a black patient with mild beta-thalassemia (beta-thal) major due to a homozygosity for the C----T mutation at position -88 of the beta promoter and with a high Hb F level. Sequence variations in the 5'HS-2 were the same as observed for the beta s chromosome with haplotype number 3, while most of the G gamma promoter and the A gamma promoter had sequences similar to that of the beta S chromosome with haplotype number 19. This atypical haplotype (number 19A) is apparently associated with an increased gamma chain production which is particularly evident during periods of severe hematopoietic stress.