[Quality control of labile blood products. Why and howto properly take a specimen? Labile Blood Products Group of the French Blood Transfusion Society]. / Le contrôle de qualité des produits sanguins labiles. Pourquoi et comment bien prélever un échantillon? Groupe de produits sanguins labiles de la Société française de transfusion sanguine.

This article reviews the various techniques of sampling used for the quality control of blood cell products. The importance of this stage for the validity of quality control results is emphasized. Three sampling methods, i.e., stripping, the sterile connection of sampling bag and the destructive method, are described in the form of operating modes and analyzed according to their advantages and drawbacks. The results of a comparative study carried out by the working group 'Blood Cell Products' of the French Society of Blood Transfusion are presented, showing that each method is valid and permits one to obtain a representative sample of the product to be controlled. Thus the diversity of the sampling methods allows us to select the one most adapted to the product to be controlled and to the analyses to be carried out afterward.

Four-step high-dose sequential chemotherapy with hematopoietic progenitor-cell support as induction treatment for patients with solid tumors.

BACKGROUND: Despite recent progress in modern chemotherapy, metastatic solid tumors still have a poor outcome. The delivery of increased dose intensities of cytotoxic agents could improve response rates. We assessed the feasibility and safety of a high-dose sequential chemotherapy program in chemotherapy-naive patients with solid tumors. PATIENTS AND METHODS: Thirty patients (14 with carcinoma of unknown primary site, seven with metastatic breast cancer, six with small-cell lung cancer, and three with other diseases) were treated by an induction therapy regimen consisting of four cycles of high-dose chemotherapy with hematopoietic progenitor cell and growth factor support. Peripheral blood progenitor cells were collected by apheresis as the leukocyte counts recovered from the nadir induced by the first cycle of chemotherapy (doxorubicin 75 mg/m2, cyclophosphamide 6000 mg/m2). Patients then received two cycles of etoposide (800 mg/m2) and carboplatin (900 mg/m2) separated by one cycle of doxorubicin (75 mg/m2) and cyclophosphamide (3000 mg/m2). G-CSF (5 microg/kg/d) was given until engraftment. Cycles were scheduled to be delivered every three weeks. RESULTS: A total of 108 cycles of chemotherapy were administered. Six patients went off study before the end of the program (three because of progressive disease, three because of toxicity). After the first cycle, a median number of 10 x 10(6)/kg CD34+ cells (range 8-30) were collected. The median number of apheresis procedures was 1 (range 1-3). From cycle 2 to cycle 4, the median number of days when there was an absolute neutrophil count of less than 500/microl increased from three to five, and the median number of days when the platelet count was less than 25,000/microl increased from three to six. Episodes of febrile neutropenia occurred in 36%, 50% and 46% of cycles during cycles 2, 3 and 4, respectively. The median numbers of days between cycle 1 and cycle 2, cycle 2 and cycle 3, cycle 3 and cycle 4 were 24 (range 20-30), 22 (range 20-36) and 22 (range 18-35), respectively. There were no treatment-related deaths. Non-hematologic toxicity included severe (WHO grades 3 or 4) nausea/vomiting in 19 (18%) cycles, mucositis in 8 (7%) cycles and diarrhea in 7 (6%) cycles. CONCLUSION: Support with hematopoietic progenitor cells and growth factors allows the timely administration of repetitive cycles of high-dose chemotherapy in chemotherapy-naive patients, resulting in a significant increase in dose intensity. Toxicity is noteworthy but manageable and does not compromise further therapy.

[Paraneoplastic edema syndromes in acute myelomonocytic leukemia: role of TNF-alpha and IL-6?]. / Syndrome oedémateux paranéoplasique au cours d'une leucémie aiguë myélomonocytaire: rôle du TNF alpha et de l'IL-6?

We report the case of a 56 year-old man in remission of a Hodgkin's disease who had an acute myelomonocytic leukemia with major edemas. Chemotherapy temporarily allowed a concomitant regression of edemas, hyperleukocytosis and tumor necrosis factor and interleukin-6 levels which were initially elevated. We discuss the role of these two cytokines in endothelium permeability disorders.

[Ultradian variations of interleukin-1 beta concentration (IL-1 beta), interferon gamma (IFN-gamma) and soluble IL-2 receptors (RIL-2) in healthy volunteers humen]. / Variations ultradiennes des concentrations en interleukine-1 beta (IL-1 beta), interféron-gamma (IFN-gamma) et récepteur soluble de l'IL-2 (RIL-2) chez l'homme volontaire sain.

The aim of this study consisted in underlining the concentrations' evolution in time of two cytokines: IL-1 beta and IFN-gamma, as well as that of RIL-2 (soluble receptor for IL-2), in healthy volunteers. Under normal physiological conditions, these molecules are secreted in small amounts, according to a sequential chronology. Thus, 25 blood samples were carried out from each of the four subjects included in the study, one every 20 minutes, during 8 hours. Concentrations were determined with the ELISA method. Spectral analysis was carried out using the Frequencies Spectrum method (Fourier Transform of a temporal series). This method allows a rhythm search over a milited number of values, in a short interval of time, provided having enough samples. Important variabilities have been observed between all subjects sera, particularly for IL-1 beta (Range (mean +/- SD): 132 +/- 34.6 pg/ml to 250 +/- 17.5 pg/ml, one subject had low values: 2.34 +/- 0.53 pg/ml) and RIL-2 (Range: 677 +/- 175 pg/ml to 1,187 +/- 46.2 pg/ml). Furthermore, ultradian serum variations, characterized by the relation ((Cmax-Cmin)/Cmax) x 100, have been assessed for each subject and each molecule: IL-1 beta (Range: 18.9 to 92.2%), IFN-gamma (Range: 66.3 to 74.2%) and RIL-2 (Range: 11.6 to 86.3%); they might be the manifestation of a periodicity; this hypothesis has been confirmed by the spectral analysis. Thus, the concentrations' variation with time showed a periodicity (Range: 80 mn to 240 mn, the most frequent period was 160 mn (2 h 40 mn)).

Acute monocytic leukemia with B cell markers expression following B chronic lymphocytic leukemia.

A case of chronic lymphocytic leukemia (CLL) treated with chlorambucil, followed by the development of an acute monoblastic leukemia, is described. Cytofluorometric quantitative immunophenotype was determined during the blastic phase. Whereas small lymphocytes displayed a CD19+; CD24+; CD37+; CD5+ phenotype, the blastic population exhibited, besides CD13, CD14 and CD15 positivity, which is usually noted in such a monoblastic leukemia, definite CD9, CD10, CD22, CD24, CD37, CD5 and CD4 staining. Such results argue against a complete independence between the two clones, although their similarity could not be demonstrated.

Immunocytochemical identification of nuclear structures containing snRNPs in isolated rat liver cells.

Small nuclear ribonucleoproteins (snRNPs) containing U1, U2, U4, U5, and U6 small nuclear RNAs were detected by ultrastructural immunocytochemistry in the nuclei of isolated rat hepatocytes using Fab fragments of anti-Sm and anti-RNP autoantibodies. Their localization was carried out in normal cells and in cells treated with two drugs, the adenosine analog DRB and CdCl2, which alter the number and distribution of nuclear RNP components. It was found that more precise determination of the distribution of these small RNAs could be obtained by using two complementary procedures in parallel rather than either one alone. They consisted of an indirect immunoperoxidase labeling carried out before embedment and an indirect immunogold labeling applied to thin sections of cells embedded in Lowicryl K4M. The results indicate that snRNPs are associated with all extranucleolar perichromatin fibrils and granules and interchromatin fibrils, which confirms that they occur in structures involved in the synthesis and processing of hnRNA. The snRNPs are not associated with nucleolar perichromatin granules induced by DRB, which confirms that there may be two kinds of perichromatin granules. The snRNPs are also associated with the still enigmatic interchromatin granules which apparently do not contain hnRNA but at least in DRB-treated cells, also contain ribosomal RNA.

Relationship between snRNA species contained in nuclear antigens recognized by autoantibodies and the clinical profile in systemic rheumatic diseases.

Antibodies to extractable nuclear antigens (ENA) are generally used in the diagnosis of connective tissue diseases. Using a rapid, very sensitive method we have shown that extractable nuclear antigens, which are now well-characterized at the molecular level, differ by their RNA content. The method was applied to the sera of 17 patients suffering from different connective tissue diseases. The results show that mixed connective tissue disease (MCTD) and other mild connective tissue diseases are characterized by the presence in the antigen of U1 small nuclear RNA (U1 snRNA) only. On the other hand, antibodies from 6 out of 8 patients tested with Systemic Lupus Erythematosus (SLE) recognize antigens exhibiting a more complex RNA pattern. Three of them precipitated all five snRNAs U2, U1, U4, U5, U6 whereas some snRNAs were lacking or quantitatively less important in precipitates obtained with the three others.

Primary structure identification of snRNAs present in highly purified snRNPs from HeLa cells.

Extensive purification of snRNPs as a subset of hnRNP from HeLa cells has been previously reported (Brunel et al. (1981), Nucleic Acids Research, 9, 815). These snRNPs were shown to contain discrete RNA species comigrating in gel electrophoresis with authentic U1, U2, U4, U5 and U6 species. We now report sequence analysis data of about 50 nucleotides from the 3'-end which serve to positively establish the identity of snRNAs present in these purified snRNPs. Sequence heterogeneity was found at the 3'-end of U4 species. A minor species identical to U1 at its 3'-end but slightly shorter was identified as the U1 described by Lerner et al. (Nature (1980) 283, 220-224) through sequencing of the 5'-end. When unfixed hnRNP are centrifuged in a CsCl gradient containing 4M guanidinium chloride instead of 0.5% sarkosyl as above, a band containing only one RNA species was observed. T1 RNAse fingerprinting and sequence analysis of the oligonucleotides produced allowed identification of this RNA as U5 snRNA.