The peopling of Madeira Archipelago (Portugal) according to HLA genes.

The Madeira-Porto Santo Archipelago was officially colonized in 1420 by Portuguese settlers. Its importance in Columbus' information for the American discovery and for slave traffic across the Atlantic is unquestionable. Thus, a complex peopling may have given rise to a present-day high admixture of ethnicities according to HLA genes. A sample of 173 healthy unrelated Madeirans was analysed and compared with 6986 HLA chromosomes from other worldwide populations. Genetic distances, neighbour-joining dendrograms and correspondence analyses were used for comparisons. Southern European, North African (including Canary Islands), Jewish and Mediterranean typical HLA alleles were found and genetic distances from Madeirans to these populations were the closest ones. In addition A*24-B*65-DRB1*0102-DQB1*0501 and A*68-B*08-DRB1*0301-DQB1*0201 haplotypes were newly found in Madeira and not found in any other population. Jewish-Armenian-Middle East haplotype (A*33-B*65-DRB1*0102-DQB1*0501) is one of the most common haplotypes; this haplotype is also present in Spaniards and North Africans. Quantitatively, Portuguese, North Africans (Algerians), Spaniards and Canary Islanders (in this order) are the most important parental populations to Madeirans. Results are discussed on the basis of the recorded historical peopling which does not show a noticeable African gene input in present-day Madeiran population according to our data; one of the closest related populations found is the Canary Islanders, suggesting that Guanche (Canary Islands first inhabitants) slaves gene flow is still noticed at present, both in Madeira and in Canary Islands populations.

HLA genes in Uros from Titikaka Lake, Peru: origin and relationship with other Amerindians and worldwide populations.

Uros population from the Titikaka Lake live in about 42 floating reed ('totora') islands in front of Puno City (Peru) at a 4000 m high altiplano. They present both an mtDNA and a human leucocyte antigen (HLA) profile different from the surrounding populations: mtDNA A2 haplogroup is common to Uros and Amazon forest lowland Amerindians. HLA genetic distances between populations have been calculated and neighbour-joining dendrograms and correspondence analyses were carried out. Approximately 15 006 HLA chromosomes from worldwide populations have been used for comparisons. Only eight HLA-A alleles have been found, three of them accounting for most of the frequencies. The same phenomenon is seen for HLA-B, HLA-DRB1 and HLA-DQB1 alleles: a few alleles (3, 4 and 3, respectively) are present in most individuals. The presence of HLA-B*4801 and HLA-DRB1*0901 alleles in a relatively high frequency (although not the most frequent alleles found) is a characteristic shared with Asians and some populations from the Andean altiplano. Three specific Uros haplotypes have been found among the most frequent ones: HLA-A*680102-B*3505-DRB1*0403-DQB1*0302; HLA-A*2402-B*1504-DRB1*1402-DQB1*0301; and HLA-A*2402-B*4801-DRB1*0403-DQB1*0302. The present study suggests that Uros may have been one of the first populations from the shores of the Titikaka Lake coming from the Amazonian forest, which might have given rise to other later differentiated ethnic group (i.e. Aymaras). Uros HLA profile is also useful to study genetic epidemiology of diseases linked to HLA and to construct a future transplant waiting list by adding up regional lists in order to get a bigger pool for transplanting with better HLA matching.

ADAMTS-13 activity and von Willebrand factor levels in methylene-blue photo-inactivated plasma processed by either the Springe method or an 'in house' system.

BACKGROUND AND OBJECTIVES: Methylene blue photo-inactivated plasma (MBPIP) has been reported to be less effective than fresh-frozen plasma (FFP) in the treatment of thrombotic thrombocytopenic purpura, which suggests a reduced content of the von Willebrand factor metalloprotease ADAMTS-13 in MBPIP. MATERIALS AND METHODS: ADAMTS-13 activity and von Willebrand factor antigen (vWF:Ag) levels were measured in plasma before and after photo-oxidation by either the Springe method or a commercial 'in house' system as well as in cryoprecipitate-poor plasma (CPP) and FFP (20 units each). RESULTS: Levels of ADAMTS-13 activity in MBPIP processed by the Springe method or the commercial 'in house' system were comparable to one another and did not significantly differ from levels found in FFP [median (range): 114% (57-139%), 99% (74-123%), and 106% (70-130%), respectively]. ADAMTS-13 activity was significantly reduced in CPP [median (range): 87% (70-107%) as compared with FFP (P < 0.05). Levels of vWF:Ag decreased after photo-oxidation by both methods. CONCLUSION: In vitro ADAMTS-13 activity was conserved in MBPIP processed by the two photo-oxidation methods analysed and did not significantly differ from levels found in FFP.

An active haemovigilance programme characterizing the safety profile of 7437 platelet transfusions prepared with amotosalen photochemical treatment.

BACKGROUND: An active haemovigilance programme was implemented to survey adverse events (AE) associated with transfusion of platelets photochemically treated with amotosalen and ultraviolet A (PCT-PLT). The results of 5106 transfusions have already been reported. Here we report the results of an additional 7437 PCT-PLT transfusions. METHODS: The focus of this ongoing haemovigilance programme is to document all AEs associated with PCT-PLT transfusion. Data collected for AEs include: time of event after starting transfusion, clinical descriptions, vital signs, results from radiographs and bacterial cultures, event severity (Grade 0-4) and causal relationship to PCT-PLT transfusion. RESULTS: One thousand four hundred patients (mean 60 years, range 1-96) received PCT-PLT transfusions. The majority of the patients (53.4%) had haematology-oncology diseases and required conventional chemotherapy (44.8%) or stem cell transplantation (8.6%). Sixty-eight PCT-PLT transfusions were associated with AE. Acute transfusion reactions (ATR), classified as an AE possibly related, probably related, or related to PCT-PLT transfusions were infrequent (n = 55, 55/7437 = 0.7%) and most were of Grade 1 severity. Thirty-nine patients (39/1400 = 2.8%) experienced one or more ATRs. The most frequently reported signs/symptoms were chills, fever, urticaria, dyspnoea, nausea and vomiting. Five AEs were considered severe (> or = Grade 2); however, no causal relationship to PCT-PLT transfusion was found. Repeated exposure to PCT-PLT did not increase the likelihood of an ATR. No cases of transfusion-related acute lung injury and no deaths due to PCT-PLT transfusions were reported. CONCLUSIONS: Routine transfusion of PCT-PLT is well-tolerated in a wide range of patients. ATRs related to PCT-PLT transfusion were infrequent and most were of mild severity.

Uses of plasma in Spain.

In Spain, fresh frozen plasma (FFP) currently recovered either by whole blood centrifugation or by apheresis is mainly considered as a source of plasma derivates rather than a product to be transfused. Upon this consideration, the amount of plasma transfused in the last two decades has remained stable, while the production of FFP has grown steadily during all these years. Thus, much more plasma has been derived to industry for manufacturing. Although, since 1993 a consensus conference established the clinical situation where plasma has demonstrated its efficacy, the true situation is that many indications seem not to be supported on a scientific evidence basis. Only a few studies have been performed in the last years to assess the appropriateness of these indications. We present the initial result of an ongoing survey addressed by the Madrid Blood Transfusion Centre. Based on the criteria of total amount of RBC transfused per year, large hospitals (more than 10,000 units of RBC) transfused an average of 23.87% of FFP, while medium hospitals (5000-10,000 units of RBC) used 19.5% and small ones (less than 5000) about 12.5%. It is important to point out that inside each group there were some important differences in ratio values for similar hospitals. This could indicate that much more is necessary to cope with indications. Although national figures of uses of FFP, whether in ratio or absolute terms, show a moderate consumption in comparison with published figures of other European countries, there can be no doubt that plasma overuses still seem to be present.

Infusion of lymphocytes obtained from a donor immunised with the paraprotein idiotype as a treatment in a relapsed myeloma.

A 48-year-old patient with IgA k multiple myeloma received a BMT from his HLA-matched sibling. After transplantation, the disease relapsed. Melphalan therapy followed by reinfusion of haemopoietic blood stem cells collected from the patient led to the improvement of the clinical status, although mixed chimerism and an elevated serum IgA persisted. Successful donor immunisation against an immunogenic preparation of the recipient monoclonal protein was performed before the infusion of donor T lymphocytes (DLI) into the patient. Ten weeks after the lymphocyte infusions, no monoclonal band was evidenced and donor complete chimerism was detected. The patient did not develop GVHD. Once complete remission was achieved, the idiotype vaccine was administered to the patient. Nineteen months after DLI, the patient remains in remission. Bone Marrow Transplantation (2000).

Histological changes in glioblastoma after intratumoral injection of autologous lymphocytes and human lymphoblastoid interferon.

Seven glioblastomas were studied between 1 and 6 months after intratumoral injection of autologous lymphocytes and human lymphoblastoid interferon. Morphological study showed a great number of lymphocytes within the tumor tissue, and interactions between lymphocytes and glioblastoma cells, suggesting a killing phenomenon. These data support the potential usefulness of adoptive immunotherapy in patients with glioblastoma by means of intratumoral administration of activated lymphoid cells.

[Changes in total and differential leukocyte counts during heart surgery with extracorporeal circulation]. / Variación en el recuento y fórmula leucocitarios durante la cirugía cardíaca con circulación extracorpórea.

The changes in leukocyte overall and differential counts during anesthesia and surgery were evaluated in 24 patients scheduled for cardiac surgery with cardiopulmonary bypass. All the end of cardiopulmonary bypass a marked and sustained leukocytosis was found (10.2 and 11.1 x 10(9).l-1), which was significantly different from baseline values (6.7 x 10(9).l-1), the values previous to cardiopulmonary bypass (6.2 and 6.4 x 10(9).l-1, and the values 30 minutes after it (5.3 x 10(9).l-1). In the differential count there were significant increases in neutrophils, associated with band and immature forms, corresponding with significant reductions in lymphocytes. There was no significant association of leukocyte variability during and after cardiopulmonary bypass and the perfusion time, the type of oxygenator or the need for intraoperative transfusion. All the reported changes tended to become normal during the first postoperative days.

[Evolution of antibodies and ABH plasmatic transferases in transplants with ABH difference between the recipient and donor]. / Evolución de los anticuerpos y de las transferasas plasmáticas ABH en los trasplantes con diferencia ABH entre receptor y donante.

Another important aspect of these transplants is the possibility of assessing the variations in the activity of serum ABH transferases in receptors of organs with ABH typing different from theirs. The antigenicity of these enzymes has been proven, antitransferase antibodies being found in BMT with minor mismatch, A-group receptors of O-group bone-marrow, and in SOT with major mismatch, O-group receptors of B-group liver. The study of the properties of such antibodies is of great interest in the course of GVHD or HVGD, in either form of transplant, as well as in the knowledge of the biochemical and immunologic characteristics of these ABH enzymes.

[Hospital use of fresh frozen plasma]. / Uso hospitalario del plasma fresco congelado.

PURPOSE: The aim of this work is double. On the one hand, to assess if the measures to strictly control the clinical indications of fresh-frozen plasma (FFP) transfusion may lead to a decrease of its use, and on the other to assess the importance of FFP with regard to other blood components, along with disclosing the number and characteristics of the more patients and those who receive only FFP. MATERIAL AND METHODS: Starting from data of the blood bank and the hospital records, an analysis of the use of FFP in the General Hospital was carried out, and it was correlated with the use of other blood components, mostly red cells (RC), and the hospital indices expressed as DRG. An analysis was also performed of the use of FFP in 1996 with regard to the number of transfused patients, mean consumed units in general and according to patient-groups, association with RC use and identification of high-use patients (defined as requiring over 3 FFP units). RESULTS: A decrease in the use of FFP between 1992 and 1996 was appreciated, from 1,385 to 760 units. This decrease, when correlated with the use of RC, was from 17.8 to 9.2 FFP units/ 100 RC units during this period. The FFP units/100 RC units varied from 6 to 2 in three years; this index has been stable since then. With regard to the use in 1996, 162 patients received FFP, which represents 4% of all the transfusions in the hospital. Of these, 15 patients received only plasma (9% of the patients receiving FFP and 0.3% of all transfusions). Other blood components, mainly RC, were associated to FFP in 96% of the cases. The patients consuming more FFP units were those of heart surgery and intensive care units, with significant differences with respect to others. CONCLUSIONS: This study shows a steady decrease in the use of FFP, which is stable in the last years. The patients receiving only FFP represented a low number with respect to all the patients transfused. The follow-up of these patients might provide valuable data about the benefit of adding additional security processes to standard FFP.

Presence of antibody to A- and B-transferases in minor incompatible bone marrow transplant.

The contribution of the bone marrow to plasma A- or B-transferase activities has been studied in patients who underwent incompatible bone marrow transplantation (BMT). As deduced from major incompatibility (group O recipient/A donor), the contribution of the marrow to these plasma activities was c. 5-10% of the total activity. In cases of minor incompatible transplants (A recipient/O donor), normal plasma activity was present in two patients, while no activity was found in a further two in whom a potent antitransferase was detected. The antibody inhibited both A- and B-transferase activities to a high titre. The patients in whom this antibody arose exhibited severe graft-versus-host disease.

The incidence of antibodies to low frequency antigens (LFA) in plasmapheresis donors with hyperimmune Rh antisera.

Over 50% of plasmas from plasmapheresis donors hyperimmunized for Rh antibodies were found to contain antibodies to low frequency red cell antigens (LFA). The majority of these plasmas contained antibodies to more than one LFA. On the other hand, the incidence of these antibodies in immune plasma from control plasmapheresis donors was markedly lower. The significance of the high incidence of these antibodies in grouping reagents is discussed.

Presence of an inhibitor of glycosyltransferase activity in a patient following an ABO incompatible liver transplant.

The contribution of the liver to plasma ABO glycosyltransferase activity has been studied in a group O individual transplanted with a liver from a group B donor. The B transferase activity present in the post-transplantation plasma was negligible. However, a potent B transferase inhibitor, absent from the pretransplantation plasma, was present after transplantation. The inhibitor was present in the excluded fraction following Sephadex G-25 gel filtration, but was retained by a protein A-Sepharose column, suggesting that it was an IgG antibody. This inhibitor was also effective in reducing A transferase activity.

Haemolytic anaemia to the alpha-interferon treatment: a proposed mechanism.

Auto-immune haemolytic anaemia (AIHA) has been found in a case of alpha-interferon treatment. Serum antibody and eluate were positive in the absence of the drug. Although the patient recovered after the treatment was stopped, DAGT remained positive for at least 8 months. The mechanism proposed to explain why this drug induced AIHA is similar to that proposed for alpha-methyl-dopa. Drugs could alter the red cell membrane and impair the immune system. Such changes have been observed with alpha-interferon and were related with increased autoimmunity.

Immunotherapy of glioblastoma with intratumoural administration of autologous lymphocytes and human lymphoblastoid interferon. A further clinical study.

A clinical trial of an immunotherapy which consisted of intratumoural injections of autologous lymphocytes with human lymphoblastoid interferon was evaluated in 31 patients with intracranial glioblastoma. Immunotherapy was performed after stereotactic biopsy or surgical resection. The treatment was tolerated well by all patients. Three patients showed positive response to immunotherapy as documented by transient regression or stabilization of the tumour size on computed tomography. Nevertheless, there is no significant difference in the survival time of the patients treated with immunotherapy and those not treated. We conclude that this immunotherapeutic regimen is not beneficial in patients with glioblastoma when used as single treatment after tumoural biopsy or resection.

Immune haemolytic anaemia and renal failure induced by streptomycin.

A case of acute immune haemolytic anaemia and renal failure induced by streptomycin, is reported. The clinical features are similar to those in a case previously reported in which no in vitro proof of antibodies was obtained. In this case, streptomycin-specific IgG antibodies, with both k and lambda light chains, could be demonstrated. The streptomycin bound strongly to the red cell membrane, apparently through chemical groups related to the M antigen and possibly also to the D antigen. Complement-fixation by the drug-specific IgG antibodies, after reaction with the streptomycin-coated red cells, could also be demonstrated. On the basis of these findings, our conclusion is that a complement-fixing hapten-cell mechanism was the main cause of the intravascular haemolytic episode suffered by the patient on exposure to streptomycin. This drug had been prescribed 15 years earlier for pulmonary tuberculosis and he had since injected himself with it whenever he felt "flu" symptoms, without harmful effects, until now.

Intrathecal injection of autologous leucocytes in glioblastoma: circulatory dynamics within the subarachnoid space and clinical results.

In part I of this report, the CSF circulatory dynamics of autologous leucocytes labelled with indium-111 and injected in the subarachnoid space, in patients operated on for glioblastoma, were studied. In the Part II, a series of 11 patients with recurrent glioblastoma was studied for evaluating the efficacy of intrathecal injection of autologous leucocytes. Six patients previously had radiotherapy. The results in Part I show that after intrathecal injection of autologous leucocytes, these cells follow throughout the subarachnoid space and pass to the systemic blood circulation, showing no evidence of colonization of the tumour or deposit in the tumoural region. The mean survival of the patients studied in Part II was 8 months. Those six patients who received radiotherapy had a mean survival of 11.4 months, and those five who received only intrathecal injection of autologous leucocytes after surgery, had a mean survival of 4 months. This results seem to demonstrate that immunotherapy, as used in this study, is ineffective in patients with glioblastoma.

[Determination of the cytokine level during storage of partially leukocyte-depleted (<0.5 x 10(9)) or totally leukocyte-depleted (<0.5 x 10(6)) platelets]. / Determinación del nivel de citocinas durante el almacenamiento de plaquetas parcialmente desleucocitadas (< 0.5 x 10(9)) o "totalmente" desleucocitadas (< 0.5 x 10(6)).

PURPOSE: To evaluate the levels of interleukin 1 beta (IL-1 beta), interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-alpha) during the storage of pools of PCs obtained after removing the buffy-coat, in comparison with the amount of leukocytes present in these components. MATERIAL AND METHODS: Blood was collected in quadruple-bag-system containing 63 mL of CPD as anticoagulant and 100 mL of SAG-M solution as additive for the red cells. Approximately twelve hours after collection, blood separation was made automatically by Compomat (NPBI) and platelet concentrates were prepared from the buffy-coat fraction. Five or six PCs were mixed to obtain a pool (n = 28). Eight pools were WBC reduced by filtration (PXL-8, Pall España). Before storage a sample of each pool was obtained in order to count platelets and WBC as basal values. All kind of pools were stored at room temperature with continuous agitation during a period of 10 days. Volumes were measured by weight and specific gravity. Platelets and leukocytes were counted in the Coulter-counter (STKR Counter. Izasa) or in Nageotte chamber for the filtered products. On days 1, 4 and 7 interleukins were measured by ELISA (EASIA Kits, Medgenix Diagnostics, Brussels), Lecture was done at 450 nm using a spectophotometer (ANTHOS 2001). Lower limits of sensitivity were 2 pg/mL for IL-1 beta and 3 pg/mL for IL-6 and TNF-alpha according to the manufacturer. Wilcoxon test was used for statistical analysis. RESULTS: The average volume of the pools was 460 mL and 374 for the filtered ones. The total platelet amount was 3.63 x 10(11) and 2.8 x 10(11) respectively, with a WBC contamination of 380 x 10(6) and 0.54 x 10(6) for the filtered. The yield of platelets after filtration was 84% with a loss of 99.90% of WBC (3-log). The measures of interleukins were not homogeneous, but a great variability was found shown among the different pools, not always in relationship with the amount of leukocytes. The levels of IL-1 beta were 3.58 pg/mL on day 1, 6.36 pg/mL on day 4 and 8.76 pg/mL on day 7 (p < 0.005). For the IL-6 we found 13.08 pg/mL on day 1, 15.43 pg/mL on day 4 and 19.77 on day 7 (p < 0.05). For the TNF-alpha, 13.65 pg/mL an day 1,24.33 pg/mL on day 4 and 30.10 pg/mL on day 7 (p < 0.05). In the filtered pools the detections of IL-1 beta and IL-6 were always under the sensibility threshold, but there was an increment of TNF-alpha on day 7 (16.91 pg/mL). Microbiologic cultures were always negative. CONCLUSION: The accumulation of IL-1 beta, IL-6 and TNF-alpha is not prevented by the fact of removing the buffy-coat layer when preparing PCs, although these levels are considerably lower in comparison with those obtained by the PRP technic. Filtration of pooled PCs avoids the presence of these cytokines except TNF-alpha, where a low amount can be detected.