[Hypotension and adverse transfusion reactions: from the associated clinical signs to the hypotensive transfusion reaction]. / L'hypotension artérielle dans les effets indésirables receveurs: du signe clinique associé à la réaction transfusionnelle hypotensive.

OBJECTIVE: The first aim of this study was to confirm the presence of hypotension blood transfusion reactions and to assess the part of hypotension as a principal event, as defined by the literature but not characterized in French haemovigilance data. As well, recent series of several cases led us to consider a possible incidence increase. STUDY DESIGN: Using a retrospective observation, the haemovigilance data from 2000 to the end of 2007 of two French regions were reviewed. During this period, 1159657 blood units were transfused by nearly 100 hospitals and 3727 adverse reactions observed. RESULTS: One hundred and sixty-eight adverse reactions with hypotension were noticed and analyzed, representing 4.5% of all transfusion reactions and revealing an incidence of 14.5 for 100000 blood units transfused. It turned out to be mostly male recipients, severe reactions and appearing rather in the beginning of transfusions. Although platelets having greater incidence, all types of blood products may be involved. The clinical diagnosis was the following: 40 to 47% were classified as febrile reactions, 13 to 17% were allergic reactions, 8 to 9% were due to immunologic and/or haemolytic reactions, 5 to 7% resulted of cardiologic disorders, 5% resulted of hypovolemic contexts and 22% were unexplained hypotensive transfusion reactions. CONCLUSION: In about three cases out of four, transfusion-induced hypotension was associated with other clinical reactions. Indeed, hypotensive transfusion reactions were identified, having an incidence of 3.2 for 100000 blood units transfused. Furthermore, there was no explanation found for the incidence increase in our region during 2007. A national study was suggested to analyse the national data as well as a prospective study to clear out this type of transfusion reactions.

Pregnancy in women with immune thrombocytopenic purpura.

Thirty-six women with immune thrombocytopenic purpura were studied during 37 pregnancies, and maternal characteristics with predictive value for the fetal platelet count were determined. Nine neonates were thrombocytopenic, with a platelet count of less than 50 x 10(9)/L in eight. Four of these nine neonates delivered to a subgroup of 31 mothers were studied prospectively; the frequency of thrombocytopenia in neonates of women with immune thrombocytopenic purpura was thus 13%. Only two of these nine neonates presented with hemorrhagic syndromes (two, petechial purpura; one, intracranial bleeding). The frequency of neonatal thrombocytopenia was higher in mothers with deep thrombocytopenia and in those who had not responded to corticosteroid treatment following diagnosis. No prognostic value could be assigned to the other maternal characteristics studied, such as a history of splenectomy, maternal treatment at the time of delivery, or the presence of platelet autoantibodies evaluated either with the platelet immunofluorescence test or the platelet Western blot immunoassay.

[Evaluation of the organization of pre and post transfusion screening of viral infection markers in blood recipients in France in 2001]. / Evaluation de l'organisation du dépistage sérologique pré- et post-transfusionnel en France en 2001.

In France, data collection related to blood recipient's viral infectious disease markers pre and post-transfusion is a legal requirement for hospitals. Our study aimed to evaluate the actual modalities of this extensive screening in 2001, six years after the Ministry of health issued recommendations. A questionnaire was sent to the haemovigilance correspondents in hospitals having transfused labile blood products (LBP) in 2001. A total of 1463 hospitals having transfused 85% of LBP in France responded. 82.4% of hospitals have written guidelines for pre-transfusion screening of viral markers, mainly for HIV and hepatitis C. A frozen repository storage is held by 23.9% of hospitals with storage durations between 1 to 40 years. 84% of hospitals have written guidelines for post-transfusion screening. The test prescriptions are mostly done by physicians from clinical services and they include in more than 80% of cases, HIV and HCV markers. Only 12% of hospitals recontact the patient in case of a no show. Even though 77.5% of responding hospitals have labile blood products recipients follow up processes, their effectiveness remains quite low, only 16% of recipients having test results available at the hospital.

T-cell depletion of bone marrow transplants: assessment of standard immunological methods of quantification.

The efficacy of bone marrow transplant (BMT) T-cell depletion for the prevention of acute graft-versus-host disease (GVHD) has been demonstrated in animal models and in clinical studies. The importance of T-cell depletion has to be evaluated with standardized methods suitable for routine purposes. We report herein an in vitro mature T-cell depletion using a cocktail of three monoclonal antibodies (CD2, CD5, and CD7) and baby rabbit complement in 38 histocompatibility leucocyte antigen (HLA)-identical BMT with no more than grade II acute GVHD. The T-cell depletion was quantified using three prestandardized immunological methods: immunofluorescence (IF) analysis, SRBC-rosetting assay, and PHA proliferation assay. A mean of 97.5% IF-assessed T-cell depletion was achieved in the 38 BMT. The immediate IF analysis using three distinct sets of anti-T-cell monoclonal antibodies allowed us to detect a mean of 1.2% residual T cells. The SRBC-rosetting assay was not useful to quantify T-cell depletion because no residual SRBC-rosette-forming cells could be detected in every case. The results of a prestandardized PHA-induced proliferation assay gave a mean 96.7% inhibition of proliferation, and they were correlated with the IF results although the IF threshold of detection was higher. From these data we conclude that our in vitro T-cell-depletion procedure is reproducible and that standardized simple immunological methods such as immediate immunofluorescence analysis and PHA proliferation assay provide good tools to assess a T-cell depletion effective in the prevention of acute GVHD.

Hepatitis C virus infection and allogeneic bone marrow transplantation.

Serum antibodies to hepatitis C virus (HCV) were tested for inpatients undergoing allogeneic BMT to determine the risk of acquiring HCV infection and the role of HCV in posttransplant liver complications. The HCV seroconversion rate was evaluated according to the date of BMT and blood donor screening at the time. Anti-HCV antibodies (anti-HCV) were detected with a second-generation ELISA and confirmed with a second-generation radioimmunoblot assay. All patients received leukocyte-depleted blood products and most received apheresis platelet concentrates. One hundred twenty of 181 consecutive patients transplanted from January 1987 to December 1991 were anti-HCV-negative before BMT, had at least 6 months of follow-up, and were thus evaluated for the seroconversion rate. Before screening for non-A, non-B hepatitis, 14% of the patients seroconverted to HCV (0.44% per unit transfused). After introduction of screening for alanine aminotransferase and antibodies to hepatitis B core antigen the risk of seroconversion was 4% per patient (0.26% per unit). When, in addition, blood was screened for anti-HCV the risk fell to 1.6% (0.03% per unit). Positive anti-HCV status before and after BMT was not predictive of veno-occlusive disease, liver graft-versus-host disease (GVHD), or death due to liver dysfunction. In contrast, the risk of chronic hepatitis was significantly increased.

Effects of irradiation on human platelets stored at 20 degrees C.

The morphologic and functional properties of platelets after irradiation with 2500 rads and storage, in first-generation containers, for 48 h in a liquid phase at 20 degrees C with continuous horizontal agitation have been analyzed and compared with a control group of the same platelets which were not irradiated. The preservation technique induced changes in the morphology and aggregation stimulated by ADP and collagen. However, no significant differences were found between the irradiated and non-irradiated groups. Irradiation is not a conditioning factor to add to the hazards of preserving platelets in a liquid phase.

Hemopoietic reconstitution after repeated autologous transplantation with mafosfamide-purged marrow.

Twenty-nine children (median age: 41 months) with advanced solid tumors received, as consolidation therapy, two consecutive courses of high-dose chemotherapy (HDC) followed by mafosfamide-purged autologous marrow transplantation (ABMT) with a 3- to 4-month interval between each course. The malignancies were neuroblastoma (n = 22), Ewing's sarcoma (n = 5) and rhabdomyosarcoma (n = 2). Patients received a preparatory regimen consisting of combined high-dose melphalan before each ABMT, with the exception of five patients who received busulfan and cyclophosphamide before the second ABMT. Prior to HDC, bone marrow sufficient for two transplantations was harvested in remission, treated with mafosfamide (50 micrograms/ml) and cryopreserved. Following incubation with the drug, a consistent inhibition (greater than 99%) of granulocyte and macrophage colony-forming units was observed. Despite the elimination of measurable hematopoietic progenitors, all patients underwent engraftment within a similar period of time after the first and the second ABMT. However, peripheral leukocyte and granulocyte recovery was delayed (median 26 and 28 days, respectively, after the first graft; median 27 and 28 days after the second graft). No difference was observed in the bacterial infections following the first and second ABMT. One patient died after the second transplant with diffuse aspergillosis. Recovery to 50 x 10(9) platelets/l occurred after a median 42 days (range 19-71) after the first ABMT and 43 days (range 14-110) after the second. Two patients died of recurrent disease before attaining a normal platelet level. One patient remained thrombocytopenic and died from visceral failure at day 200. These results demonstrate the feasibility of repeated ABMT with mafosfamide-treated marrow.(ABSTRACT TRUNCATED AT 250 WORDS)

A simple, efficient washing procedure for cryopreserved human hematopoietic stem cells prior to reinfusion.

We describe our experience with a washing procedure used on cryopreserved, thawed bone marrow (BM) and peripheral blood stem cell (PBSC) grafts prior to autologous transplantation in 50 and 12 patients respectively. The procedure consists of a stepwise dilution with 2% human serum albumin and centrifugation performed either manually or using a blood cell processor (Cobe 2991). In vitro studies showed mean recoveries of 80.8% for BM nucleated cells and 73.9% for BM hematopoietic progenitors (CFU-GM). The corresponding recoveries for PBSC were 89.1 and 93.9%. After 4 h storage at +20 degrees C of the manipulated grafts, no significant loss of CFU-GM was observed. We conclude that the technique is simple and efficient for washing large numbers of hematopoietic stem cells. This method may avoid the clinical complications often arising with unwashed grafts.

Successful engraftment after autologous transplantation of 10-day cultured bone marrow activated by interleukin 2 in patients with acute lymphoblastic leukemia.

Disease recurrence remains the major problem in autologous bone marrow transplantation (BMT) for hematologic malignancies. To improve the therapeutic efficiency of autologous BMT, we investigated the use of autologous marrow activated in vitro with interleukin 2 (IL-2) to generate killer cells for in vivo purging. A feasibility trial was initiated in 5 patients with poor prognosis acute lymphoblastic leukemia, who were transplanted, after marrow ablative therapy, with autologous marrow cultured for 10 days with 10(3) units of IL-2/ml. A highly significant increase in NK activity and an induction of LAK activity were observed after incubation. Patients received 0.64 to 1.56 X 10(8) cultured BM cells/kg and 1.87 to 44.8 x 10(4) CFU-GM/kg. Four patients engrafted and achieved granulocyte counts > 0.5 x 10(9)/l on days 35, 24, 36 and 22 after transplant. Three of these patients showed platelet recovery to > 50 x10(9)/l on days 25, 42 and 40 after transplant. One patient remained thrombocytopenic until relapse. One patient died on day 12 after transplant. This study demonstrates that cultured BM activated with IL-2 can be used successfully for hematological rescue in the clinical setting.

Effect of human granulocyte colony stimulating factor on neuroblastoma cell growth.

We have studied the effect of recombinant human granulocyte colony-stimulating factor (G-CSF) on the growth of three different human neuroblastoma (NB) cell lines, using a clonal cell culture and a short proliferation test. The results have shown a lack of effect of G-CSF on the three cell lines. These data promote the use, in clinical trials, of G-CSF as adjunctive treatment in children with NB receiving intensive chemotherapy.

Successful infusion of 40 cryopreserved autologous bone-marrows. In vitro studies of the freezing procedure.

Cryopreservation of human bone marrow may be helpful to use supralethal chemoradiotherapy in order to cure malignant diseases. We report here a cryopreservation procedure from large volumes of human bone marrow which can be applied in routinal use. Whole bone marrow in 10% Dimethylsulfoxide (ME2SO) was frozen at an 1 degree C/min. controlled rate and was stored into liquid nitrogen. After thawing and before infusion, both ME2SO and free hemoglobin were removed. The in vitro recovery of nucleated cells and the myeloid stem cell assay (CFC) were performed as quality control. About 60% of the marrow cells and 40% of the total CFC number were recovered at the end of the procedure. Using this technique, 40 patients (25 children with malignant lymphoma and 15 adults with lymphoma and solid tumors) were transplanted with autologous cryopreserved bone marrow after receiving intensive chemotherapy. Three of them received both chemotherapy and a total body irradiation. Engraftment was achieved in all patients. Rise in leucocyte count (greater than 1.10(9)/l) occurred within an average of 17 days. In conclusion, in autologous bone marrow transplantation, this method of cryopreservation is effective to obtain rapid hematological reconstitution in patients treated for malignant diseases by intensive cytoreductive regimens.

[Evaluation of pre- and post-transfusion serological screening in France in 2001]. / Evaluation du dépistage sérologique pré- et post-transfusionnel en France en 2001.

Screening of labile blood products recipients for HIV and HCV has been performed in France since a government recommendation was issued in 1996. It has been designed to get transfusion related contamination of recipients through pre- and post-transfusion serological tests. Since then, residual risk has decreased dramatically and it was suspected that recommendations might sometimes be ignored. A nationwide survey has been done to measure the real screening rate and its cost efficacy ratio. In addition accuracy of tracability and recipients mortality has also been evaluated. A random sample of 1115 labile blood products among all the 1203 378 distributed during first semester of 2001 in France has been drawn. They have been matched with test results obtained in hospital files. Tracability has been considered accurate if name, surname and birth date of recipients were exactly the same both in hospital file and in the file of the Etablissement Français du Sang. A total of 1092 hospital files has been retrieved. Pre transfusion HIV and HCV tests have been performed in 58.5 % of cases, 95 % CI [55.6-61.5], and post-transfusion tests in 30.5 % [28.5-35.5] of cases. Only 19.5% [16.6-22.6] of recipients, not known to be dead 6 months after transfusion, have had both pre and post-transfusion tests. No HIV or HCV contamination has been notified by the Haemovigilance network during the same period. Accuracy rate of tracability was 96.25% [94.9-97.3]. Furthermore 35.8% [33-38.7] of recipients were found dead within 6 months after transfusion. A logistic regression analysis showed that the hospital area, the hospital size (more than 300 beds) and the annual amount of blood bags transfused in it (less than 5000) were factors independently associated with having a full pre and post-transfusion screening. Currently, the screening program may detect 0.14 cases of HIV and 0.05 HCV transfusion related contamination of recipient every year. The total cost of this program is about 20 million euro and the cost per case exceeds 110 million euro. The program will be of no use in case of an emerging transmitable disease. This program does not comply to any evaluation criteria of screening programs and its cost efficacy ratio is very poor.

[Regional program to improve the quality of blood transfusions in hospitals: experience of the Poitou-Charentes region]. / Programme régional d'amélioration de la qualité transfusionnelle dans les établissements de santé: expérience de la région Poitou-Charentes.

In the Poitou-Charentes area, a regional pilot program was implemented over a two year-period to improve transfusion safety in public and private hospitals. This program consisted in: (i) an evaluation of the transfusion chain in hospitals; (ii) a regional program to guide hospitals in improving the quality process. Five workgroups were set up. Three persons in each hospital should participate in the workgroup: one representing the administration, one the medical staff and one the nursing staff. After a six months follow-up several hospitals were prompted to implement corrective and preventive measures to improve transfusion safety; (iii) a letter was regularly published to contribute to set-up a regional haemovigilance network. Such a quality improvement program revealed to be a relevant method to steer the changing blood transfusion process in hospitals.

[Transfusion and alloimmunization in sickle cell anemia patients]. / Transfusion et alloimmunisation chez les patients drépanocytaires.

Transfusion therapy for sickle cell anemia is limited by the development of antibodies to red cell antigens. The aim of this study was to evaluate whether transfusion of blood matched for antigens Rh and Kell would reduce the incidence of alloimmunization. We determined the transfusion history, red cell phenotype and development of alloantibodies in 173 patients with sickle all anemia who received transfusions. Forty nine patients were transfused exclusively with frozen red blood cells (RBL) matched for antigens Rh and Kell; the rate of alloimmunization was 8.2%; antibodies to the Jkb, Jka, Fya and S were developed; 1 patient developed 2 antibodies. In a control group of 124 patients who received standard red blood cells, the rate of alloimmunization was significantly increased to 30.6% (p < 0.05); antibodies against C, E, K, Fya were the most frequently developed and 19 patients (16%) developed antibodies reacting with different antigens. In the 2 groups, alloimmunization occurred after receiving a significantly different number of transfusions: mean 9 in the patients transfused with matched RBC and 32 in the control group. The influence of the kinetics of transfusion was not demonstrated. To assess the effect that racial differences might have on alloimmunization, comparison of the red cell phenotype of patients with that of a panel of unselected blood bank donors was performed: the patients had a significant decrease in the frequency of red cell antigens corresponding to most of the detected alloantibodies JkB, C, S. Fyb, Fya and Kell.(ABSTRACT TRUNCATED AT 250 WORDS)

[Post-transfusion purpura. An unknown cause of acute immune thrombocytopenia. 4 new cases]. / Le purpura post-transfusionnel. Une cause méconnue de thrombopénie aiguë immunologique. Quatre nouveaux cas.

Post-transfusion purpura is characterized by the occurrence of acute immune thrombocytopenia 5 to 10 days after transfusion of platelet-containing blood products in subjects who had been alloimmunized to specific platelet antigens. Four cases are reported here. Three of these 4 patients, who had a rare PLA1 platelet phenotype, had developed, during a previous sensitization (pregnancy n = 2, transfusion n = 1), an allo antibody directed against PLA1 antigen. The fourth patient presented a specific anti-PLA2 antibody. Thrombocytopenia (platelet count between 4 and 40 x 10(9)/1) appeared 1 to 12 days after the responsible transfusion and showed as haematomas (n = 3) or haemoptysis (n = 1). One patient was asymptomatic and remained untreated. The remaining 3 patients received corticosteroids orally associated, in one case, with infusions of human immunoglobulin concentrates. Thrombocytopenia was corrected within 5 to 13 days. In such cases, whenever another blood transfusion is necessary, a preventive treatment with perfectly platelet-free blood products is mandatory. Platelet depletion by freezing-thawing of red cell concentrates is probably not always sufficient, since recurrence was observed with such a product in one of our patients. The exact cause of immune destruction of autologous platelet remains a mystery.

[The decrease of preoperative autologous transfusion in France has not been linked to an increase of homologous red cell concentrates]. / La diminution de l'autotransfusion programmée en France ne s'est pas accompagnée d'une augmentation de consommation de concentrés de globules rouges homologues.

OBJECTIVE: The use of preoperative autologous blood transfusion has dramatically decreased in France. The aim of this study was to evaluate the evolution of practice both of autologous and homologous Red Blood Cells (RBC) concentrates transfusion between 2002 and 2005, and to asses the consequences of the highlighted changes. MATERIAL AND METHODS: Data on blood transfusion are collected and validated nationwide by a network of regional coordinators of haemovigilance. For each hospital, from 2002 to 2005, the annual changes in the number of transfused homologous and autologous RBC have been evaluated. The ratio of preoperative autologous RBC, number of autologous RBC divided by the number of all RBC, has also been calculated. Hospitals have been split into three cohorts, according to their 2005 autologous RBC ratio. For each cohort, correlations between the variations of the number of autologous, homologous and total RBC in each hospital have been studied. RESULTS: Data have been validated for 22 French regions that performed 71.8% of the total French transfusion in 2004. In 2005, 1,831,544 labile blood products have been transfused in 1197 hospitals and clinics among which 379 have used preoperative autologous transfusion. A total of 37,289 autologous RBC have been transfused in 2003, 28,689 in 2004 (-23.1%) and 17,758 in 2005 (-38.1%). The first cohort of 269 hospitals had a ratio of autologous RBC under 3%, the second cohort of 38 hospitals, a ratio between 3 and 6%, while the third cohort of 72 hospitals had transfused 6% or more of autologous RBC. In the two first cohorts, non-surgical activities were so large that it was impossible to assess the changes in surgical use of transfusion. The third cohort, with a ratio of 6% or more, was essentially devoted to surgery (88% of beds). These hospitals and clinics have transfused 13,076 autologous RBC in 2002 and 8583 in 2005 (-34.4%). In this group, there was a statistical correlation between the decrease of autologous RBC and the decrease of total RBC (r(2)=0.36), and no increase in the transfusion of homologous RBC has been observed. During the same period, neither hospitals nor clinics showed any decrease of their surgical activity. The drop of autologous RBC transfusion led to a decrease of the total number of RBC transfused and thus, to a decrease of the global exposure to transfusion hazard. CONCLUSION: The present results confirmed a decline of preoperative autologous transfusion in France, between 2002 and 2005. Meanwhile no supplementary need of homologous RBC has been observed among hospitals, performing surgery that formerly had a high ratio of autologous RBC.

[Value of filariopheresis in the treatment of Loa loa filariasis]. / Intérêt de la filariophérèse dans le traitement de la filariose A Loa loa.

To prevent the side effects as encephalitis, related with diethylcarbamazine treatment in loiasis, an exchange blood transfusion was proposed. To avoid the disadvantage of this heavy technique, the authors are proposing filariopheresis to trap the microfilariae using cytapheresis technique and filtration. They obtained a medium 75 p. cent microfilariae extraction: 55-70 p. cent by cytapheresis and 5-20 p. cent by filtration. The number of blood cells must be supervised as an important decrease of the platelets may occur temporary. This simple and not expensive technique is efficient, but diethylcarbamazine treatment must be instituted very quickly after filariopheresis session. Furthermore, the millions of microfilariae collected can provide useful antigen for immuno-parasitological tests.

Apheresis experience with a continuous flow cell separator.

The Dideco Vivacell separator is a continuous-flow centrifugation system that has only recently been used for cytapheresis. The authors' experience with this separator in 451 plateletpheresis and 164 leukapheresis procedures is presented. Platelet collection provided high platelet yields (9.53 +/- 2.85 X 10(11) with a collection efficiency of 74 +/- 14 percent for about 6 liters of total blood processed. Functional integrity was confirmed by normal in vitro tests (aggregation and response to hypotonic stress) and good in vivo recovery (55%). In leukapheresis, white cell yields were high (3.42 +/- 1.2 X 10(10) with 85 percent polymorphonuclear neutrophil cells. Their oxidative metabolism functions (generation of free oxygen radicals), investigated by chemiluminescence, were increased over donor values. Donor reactions, all of the mild citrate type, were rare.

Peripheral blood stem cell collection with a blood cell separator.

Forty-three patients with malignant nonmyeloid diseases underwent peripheral blood stem cell collections on an apheresis system (Spectra, COBE BCT, Lakewood, CO). Collections took place during the white cell (WBC) recovery phase following conditioning chemotherapy. One hundred two procedures were done after chemotherapy alone, and 72 procedures after chemotherapy plus granulocyte-colony-stimulating factor (G-CSF). Four centrifugal separation factors were tested. One and one-half patient blood volumes were processed in each procedure. The mean volume of the collected component was 158 +/- 16 mL. After chemotherapy alone, the procedures provided a mean of 0.8 x 10(8) WBCs per kg and 2.3 x 10(4) colony-forming units-granulocyte macrophage (CFU-GM) per kg of recipient body weight. The mononuclear cell percentage in the components increased with the centrifugal separation factor from 85 to 96 percent. In parallel, platelet contamination increased from 2.1 to 3.8 x 10(11). The collect hematocrit ranged from 1.0 to 2.5 percent (0.01-0.025). The collection efficiency for mononuclear cells and CFU-GM also increased with the centrifugal separation factors from 52 to 70 percent for mononuclear cells and from 55 to 68 percent for CFU-GM. Collections performed after G-CSF-stimulated mobilization were characterized by a higher neutrophil contamination independent of centrifugal separation factor, which gave a mean mononuclear cell percentage of 64 percent in the collected component. The average yield for these procedures was 2 x 10(8) WBCs per kg and 28 x 10(4) CFU-GM per kg.(ABSTRACT TRUNCATED AT 250 WORDS)

[Red cell preservation by freezing at -25 degrees C]. / Conservation des hématies par congélation à -25 degr'es C.

A method of red cell preservation by freezing at -25 degrees C is described. Glycerol is added to the red blood cells in the primary polyvinyl chloride plastic collection bag to achieve a concentration of 28 pr cent (W/V). The blood cells are concentrated by centrifugation and the supernatant glycerol is discarded. Glycerolized red cells are frozen and stored at -25 degrees C for 1 to 6 months. After thawing, Sodium chloride solutions are used to wash the red cells in the IBM Blood Processor 2991. The following parameters have been investigated before freezing, after thawing and washing and after storage of red blood cells at 4 degrees C for 24 hours: --Hemoglobin level --leukocytes and platelets --amount of 2-3 DPG and ATP. Preliminary data show that the in vitro quality of erythrocytes stored at -25 degrees C is well preserved for 4 months and that this simple method can be applied to blood preservation in any Blood Center.