[The training in transfusion medicine remains deficient in the centres of Francophone sub-Saharan Africa: results of a preliminary study]. / La formation en médecine transfusionnelle reste insuffisante dans les centres d'Afrique subsaharienne francophone: résultats d'une étude préliminaire.

PURPOSE OF THE STUDY: To evaluate the needs in staff training in transfusion centres of Sub-Saharan Africa. MATERIAL AND METHODS: This preliminary study analyzed the training level of each personnel of four blood banks of Sub-Saharan Africa, their training fields, duration and training structures. RESULTS: The needs remain high in all the fields and are critical regarding the administration of blood transfusion services, equipment maintenance and clinical use of blood.

Study of coagulation function in thawed apheresis plasma for photochemical treatment by amotosalen and UVA.

BACKGROUND AND OBJECTIVES: Photochemical treatment (PCT) based on amotosalen and ultraviolet A light (UVA) demonstrated a wide range of pathogen inactivation. However, coagulation proteins are affected by this treatment. The aim of this study was to evaluate the coagulation parameters in apheresis plasma units after thawing and processing by PCT. MATERIALS AND METHODS: Thirty apheresis plasma units were rapidly frozen at

Feasibility of prevention of perinatal HIV infection by nevirapine in rural areas of the northeast Democratic Republic of Congo, 2002-2004.

The HIV NET 012 protocol for prevention of perinatal transmission of HIV, a single-dose of nevirapine given to the mother during labor and one given to the newborn, has been adopted by several developing countries. The implementation of this protocol may be difficult in African rural regions and must be evaluated. The study was carried out at the Oicha General Hospital, Democratic Republic of Congo. HIV-infected women with a gestational age > or =36 weeks were offered participation in the study during a prenatal visit. Among the 5,016 women tested during prenatal attendance from December 2002 to December 2004, 94 (1.9%, 95% confidence interval [CI]: 1.5-2.3%) were HIV-infected. Among these women, 59 (62.8%; 95% CI: 53.0-72.6%) received nevirapine and 35 (37.2%; 95% CI: 27.4-47.0%) did not receive nevirapine prophylaxis. Twenty-six (27.7%) of these women arrived fully dilated and 9 (9.5%) were not given the drug by the midwives. Among the 59 HIV-infected pregnant women who received nevirapine before delivery, 33 (55.9%, 95% CI: 43.2-68.6%) received the drug at the start of uterine contractions and 24 (40.7%; 95% CI: 28.2-53.2%) did not receive nevirapine within the recommended time before delivery. Two women (3.4%) received nevirapine, but this was not written in the logbook. Overall, among the 101 newborns, all children received a single-dose nevirapine: 48 (47.5%; 95% CI: 37.8-57.2%) of the 101 newborns received nevirapine in the 24-72 hr following birth and 52 (51.5%; 95% CI: 41.8-61.2%) just after birth when their mother did not receive nevirapine. Even for a protocol as simple as HIV prophylaxis with a single-dose nevirapine, the application in African rural areas encounters field difficulties. Further simplified procedures and health care re-organization are still needed to guarantee a full Prevention of Mother to Child Transmission coverage for every HIV pregnant woman in Africa.

Analysis of genomic instability using multiple assays in a patient with Rothmund-Thomson syndrome.

We report on a patient with Rothmund-Thomson syndrome (RTS) whose cytogenetic evaluation showed a normal karyotype with no evidence of trisomy mosaicism or chromosomal rearrangements. Cultured lymphocytes from the patient, her mother, and a control exposed to mitomycin C and diepoxybutane did not show increased sensitivity to the dialkylating agents. Unlike some previous reports, we found no evidence of a deficiency in nucleotide excision repair, as measured with the functional unscheduled DNA synthesis assay. Glycophorin A analysis of red blood cells for somatic mutation revealed suspiciously high frequencies of both allele loss and loss-and-duplication variants in the blood of the patient, a pattern consistent with observations in other RecQ-related human diseases, and evidence for clonal expansion of a mutant clone in the mother. Discrepant results in the literature may reflect true heterogeneity in the disease or the fact that a consistent set of tests has not been applied to RTS patients.

Delayed massive immune hemolysis mediated by minor ABO incompatibility after allogeneic peripheral blood progenitor cell transplantation.

BACKGROUND: Bone marrow transplantation with minor ABO incompatibility may be followed by moderate delayed hemolysis of the recipient's red cells by donor-derived ABO antibodies. This reaction may be more severe after transplantation of peripheral blood progenitor cells (PBPCs). CASE REPORT: A 16-year-old boy underwent an allogeneic PBPC transplant from his HLA-mismatched mother as treatment for acute myeloblastic leukemia that had proved resistant to induction chemotherapy. Transfusion of the unmanipulated PBPCs proceeded without any complication, despite the difference in ABO blood group (donor, O Rh-positive; recipient, A Rh-positive). On Day 7, a rapid drop in hemoglobin to 4 g per dL was observed, which was attributed to a massive hemolysis. All the recipient's group A red cells were destroyed within 36 hours. This delayed and rapidly progressive hemolytic anemia was not associated with the transfusion of the donor's plasma. Rather, the anti-A titer increased in parallel with marrow recovery, which suggested an active synthesis of these antibodies by immunocompetent cells from the donor against the recipient's red cells. The mother's anti-A titer was retrospectively found to be 2048. Her unusually high titer is probably due to prior sensitization during pregnancies. On Day 12, the patient developed grade IV graft-versus-host disease, which proved resistant to all treatments instituted and led to his death on Day 35. CONCLUSION: PBPC transplantation with minor ABO incompatibility may be associated with significant risk of massive delayed hemolysis.

Prevalence of hepatitis G virus in a haemodialysis unit.

BACKGROUND: Recently, a novel blood-borne virus has been identified and named hepatitis G virus. Transfusion is the main route of transmission. It is known that patients on maintenance dialysis are more susceptible to infections with parenterally-transmitted viruses than the general population. The aim of the present study was to determine the prevalence of hepatitis G infection in a Belgian dialysis unit. METHODS: The entire population of our dialysis unit (82 patients) was tested for the presence of hepatitis G virus (HGV) by reverse transcriptase polymerase chain reaction. History of transfusion or renal transplantation, coinfections with hepatitis B and C viruses, and serum aminotransferase levels were also tested. RESULTS: Thirteen patients (16%) were found positive for HGV-RNA. Among these patients, 69.2% were infected by the G virus alone, 15.4% were coinfected with B virus, and 15.4% with C virus. All but one patient had a history of transfusion. Ten of the thirteen infected patients (77%) had normal aminotransferase (< 30 UI/l). Three patients had elevated aminotransferase levels (23%); one was coinfected with B virus, one with C virus, and the last one had a diabetes-induced fatty liver infiltration. No liver biopsies were performed. CONCLUSIONS: It is concluded that infection with G virus is common among dialyzed patients. This high rate of infection could be related to previous transfusions, but may as well be due to nosocomial transmission. In our series, at least one patient has been contaminated by another road than transplantation or transfusion. Finally, it does not appear clearly that chronic infection with hepatitis G virus induces liver disease, as defined by elevated aminotransferase level.

Hemovigilance: clinical tolerance of solvent-detergent treated plasma.

OBJECTIVE: This study was conducted to assess retrospectively the clinical tolerance of SD treated plasma and to compare it to other labile blood products (red blood cell and platelet concentrates). METHODS: Adverse events (AEs) related to the use of blood products at the Blood Transfusion Center (BTC) are routinely collected through a formalised system of hemovigilance. All AEs reported are entered into a safety data base which was used for the study. All AEs reported during a one-year period to the BTC were retrospectively re-assessed and descriptive statistics calculated. RESULTS: 5064 units of SD treated plasma were transfused to 894 recipients during the study period at the occasion of 1553 transfusions. No AE associated to SD treatment plasma was reported during that period. In contrast, during the same period, 485 AEs associated with the use of red blood cell concentrates (RBCC) were reported in 251 patients at the occasion of 262 transfusions. 2.1% (251/11,748) of the patients transfused with RBCC experienced one or more AEs. The incidence of AEs per unit transfused was 1.3% (485/37,332), and 2.4% (485/20,460) of RBCC transfusions were associated with one or more AEs. 142 AEs associated with the use of platelet concentrate (PC) were observed in 69 patients at the occasion of 73 transfusions. 4.2% (69/1645) of patients transfused with PC experienced one or more AEs. The incidence of AEs per unit transfused was 1.1% (142/12,772), and 2.8% (142/5034) of PC transfusions were associated with one or more AEs. All reported AEs were classified and non serious. The most frequently observed AEs were fever, chills and rashes which accounted for roughly 64% of all reported AEs. CONCLUSION: As for the overall clinical tolerance of red cell and platelet concentrates, the results of this study are in complete agreement with the published literature. The study also confirms the extremely good tolerability of SD treated plasma in comparison with other labile blood products.

[Production of monoclonal antibodies specific for the ABO blood group and rhesus D antigens]. / Production d'anticorps monoclonaux spécifiques des antigènes des groupes sanguins ABO et Rhesus D.

We report, in this work, the techniques to obtain four monoclonal antibodies specific of erythrocytes antigens. Three of this antibodies, react with the ABO Blood Groupe System (A,B and AB), are produced by three mouse hybridomas (M18-2F11, M18-4F6 et M19-45D6), obtained by fusion of Sp2/0 mouse myeloma cell with spleen cell of balb/c mice immunized with human red blood cells and selected on selectif medium (Hypoxanthin, Aminoptérin, Thymidin) (HAT) and cloned by four limiting dilutions. Where as the fourth, it is an human monoclonal antibodies against Rhesus (D) produced by heterohybridomas, realized by fusion of X63 mouse myeloma cell with human B lymphocytes, from actively immunized persons by D antigen, that are purified and transformed by Epstein-Barr virus (EBV), and selected on selectif medium (HAT), presence of ouabain and then cloned by four limiting dilutions. The specificity of the antibodies produced, has been determined by direct hemagglutinin for the mouse monoclonal antibodies and artificial for the human anti-D. The determination of isotype the heavy and light chains is released by the technique immunoenzymatique (ELISA) and immunofixation has shown that mouse monoclonal antibodies belong to class IgM kappa, and human monoclonal antibodies anti-D belong to class IgG lambda.

Prestorage leukocyte reduction with in-line filtration of whole blood: evaluation of red cells and plasma storage.

OBJECTIVES: Prestorage filtration of blood components appears to be an effective method to reduce leukocyte-induced adverse reactions and other complications. To determine whether it is better to filter whole blood before component separation, we compared the efficiency of in-line filtration of whole blood with that of postseparation filtration. METHODS: Blood was collected from normal, healthy donors into either regular triple-bag containers or into whole-blood integral-filter container systems. We then compared the in vitro storage values of leukocyte-depleted red blood cell concentrates (RBCC) kept at 4 degrees C, and plasma frozen for 1 year with nonfiltered blood components as control. RESULTS: All counts of white blood cells after filtration were < 1 x 10(6) per unit. For almost all storage parameters no significant differences were found between leukocyte-reduced RBCC and control units. The plasma fibrinopeptide A values below 30 ng/ml prior to freezing indicate that filtration does not activate the coagulation factors. Furthermore, the filtration did not influence either the biological values or the coagulation factors of plasma units. CONCLUSIONS: Whole blood filtration prior to component preparation seems to offer a useful alternative technique for obtaining leukocyte-reduced RBCC and plasma.

Multi-center European evaluation of HIV testing on serum and saliva samples.

OBJECTIVE: to evaluate the reliability of HIV antibody testing on saliva. DESIGN: matched serum and saliva samples were collected from both seronegative (n = 344) and seropositive (n = 125) individuals in five European countries. Duplicate saliva samples collected with Omni-Sal devices provided by Saliva Diagnostic System (SDS) were pooled before analysis. METHODS: all samples were analyzed by Recombinant HIV1 EIA Cambridge Bioscience and 2nd generation Abbott HIV 1&2 1A80. EIA procedures were adapted for saliva testing by modification of sample dilution and/or cut-off calculation. All saliva recording positive and/or doubtful EIA results were further analyzed by Western blot as a confirmatory method. RESULTS: EIA results obtained from sera analysis from both seropositives and seronegatives allowed for calculation of the tests' sensitivity (HIV1 Biotech: 99.2%-100%; Abbott: 100%) and specificity (both tests 100%). In the series of 125 saliva samples collected from seropositives, the EIA results were as follows: with Biotech (3 negative, 3 in the grey-zone and 119 reactive) and with Abbott (1 negative, 1 in the grey-zone and 123 reactive). One saliva sample found negative by both EIA tests, although fulfilling HIV1 WB criteria of positivity, was collected from an HIV2 infected person. Out of 125 saliva samples collected from seropositives, 121 produced positive Western Blot profiles, 4 were indeterminate and 1 was found negative whereas 125/125 sera were found positive. CONCLUSION: the reliability of HIV testing of saliva is dependent on the sensitivity of EIA tests and on the criteria used for the interpretation of Western blot tests as well. Although saliva testing offers numerous advantages for epidemiological purposes, it should not be recommended for diagnosis.

The effect of argon laser trabeculoplasty on the blood-aqueous barrier and intraocular pressure in human glaucomatous eyes treated with diclofenac 0.1%.

BACKGROUND: We studied the effect of argon laser trabeculoplasty (ALT) on the blood-aqueous barrier (BAB) in 41 eyes of 41 patients with primary open-angle glaucoma, pseudoexfoliation glaucoma, or pigment dispersion glaucoma using the Fluorotron Master II. METHODS: Fluorophotometry was performed the day before ALT and on the 3rd day after surgery at 30 and 60 min after intravenous injection of 7 mg/kg body weight sodium fluorescein 10%. Intraocular pressure (IOP) was measured using Goldmann applanation tonometry on the day before surgery and at 3rd days and 1 year (mean) after ALT. Patients were treated with argon laser by one surgeon (180 degrees, 0.1 s, 50 microns 0.6-1.0 W, 56 laser burns). Eyes were randomly assigned to either diclofenac-sodium 0.1% eye drops or vehicle. Eye drops were applied six times 1 h before ALT into the operated eyes and five times daily for 3 days postoperatively. RESULTS: On the 3rd day after ALT there was significant disruption of the BAB in the placebo-treated eyes compared to the diclofenac 0.1%-treated eyes. In the placebo-treated eyes as well as in diclofenac-sodium 0.1%-treated eyes there was a significant decrease of IOP postoperatively for up to 1 year. There was no significant difference concerning the IOP reduction after 1 year. Diclofenac-sodium 0.1% eye drops significantly stabilized the BAB on the 3rd day after ALT, compared to placebo, in this model. CONCLUSION: Diclofenac-sodium 0.1% significantly stabilized the disruption of the blood-aqueous barrier on the 3rd day after ALT. Concerning the IOP-lowering effect of ALT, the postoperative application of steroids should be avoided.

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. Their complementary activity towards weak D and partial D antigens is examined.

[Post-transfusional hepatitis due to hepatitis virus C]. / Les hépatites post-transfusionnelles dues au virus C.

Cloned even before observed, C virus hepatitis seems to live out its natural life backwards. The C virus has begun to confide a part of its secrets to molecular Biology. Today's scientific data demonstrate that, for the most part, occurrences of post-transfusional hepatitis (PTH), which are neither A nor B, are the C virus. In order to lower the risk of blood transmission of the C virus, a systematic screening of HCV antibodies has been mandatory in Belgium since 1 July 1991. Epidemiological data has testified that seroprevalence among blood donors is around 0.55%. Even though screening is an efficient measure to eliminate blood units that are suspected to be contaminated, implementing molecular Biology techniques (PCR or chain polymerization reaction) extracts detection of the viral genome, independent of the presence or absence of specific antibodies. The measurement of ALT (alanine amino transferase) as a surrogate marker in all blood units is not yet mandatory even though it has been described as a practical and low cost means to reduce PTH in blood recipients. Among the human measures available to determine the selection of blood donors are pre-donation anamnesis and awareness programmes to inform potential donors of risk factors. Post-transfusional hepatitis C is a public health concern. The residual risk of contamination by blood products remains too great. Both human and serological measures have to be improved.