Distribution of indications and procedures within the framework of centers participating in the WAA apheresis registry.

The WAA apheresis registry was established in 2003 and an increasing number of centers have since then included their experience and data of their procedures. The registry now contains data of more than 74,000 apheresis procedures in more than 10,000 patients. This report shows that the indications for apheresis procedures are changing towards more oncological diagnoses and stem cell collections from patients and donors and less therapeutic apheresis procedures. In centers that continue to register, the total extent of apheresis procedures and patients treated have expanded during the latest years.

Adverse events in apheresis: An update of the WAA registry data.

Apheresis with different procedures and devices are used for a variety of indications that may have different adverse events (AEs). The aim of this study was to clarify the extent and possible reasons of various side effects based on data from a multinational registry. The WAA-apheresis registry data focus on adverse events in a total of 50846 procedures in 7142 patients (42% women). AEs were graded as mild, moderate (need for medication), severe (interruption due to the AE) or death (due to AE). More AEs occurred during the first procedures versus subsequent (8.4 and 5.5%, respectively). AEs were mild in 2.4% (due to access 54%, device 7%, hypotension 15%, tingling 8%), moderate in 3% (tingling 58%, urticaria 15%, hypotension 10%, nausea 3%), and severe in 0.4% of procedures (syncope/hypotension 32%, urticaria 17%, chills/fever 8%, arrhythmia/asystole 4.5%, nausea/vomiting 4%). Hypotension was most common if albumin was used as the replacement fluid, and urticaria when plasma was used. Arrhythmia occurred to similar extents when using plasma or albumin as replacement. In 64% of procedures with bronchospasm, plasma was part of the replacement fluid used. Severe AEs are rare. Although most reactions are mild and moderate, several side effects may be critical for the patient. We present side effects in relation to the procedures and suggest that safety is increased by regular vital sign measurements, cardiac monitoring and by having emergency equipment nearby.

A prospective, active haemovigilance study with combined cohort analysis of 19,175 transfusions of platelet components prepared with amotosalen-UVA photochemical treatment.

BACKGROUND AND OBJECTIVES: A photochemical treatment process (PCT) utilizing amotosalen and UVA light (INTERCEPT(™) Blood System) has been developed for inactivation of viruses, bacteria, parasites and leucocytes that can contaminate blood components intended for transfusion. The objective of this study was to further characterize the safety profile of INTERCEPT-treated platelet components (PCT-PLT) administered across a broad patient population. MATERIALS AND METHODS: This open-label, observational haemovigilance programme of PCT-PLT transfusions was conducted in 21 centres in 11 countries. All transfusions were monitored for adverse events within 24 h post-transfusion and for serious adverse events (SAEs) up to 7 days post-transfusion. All adverse events were assessed for severity (Grade 0-4), and causal relationship to PCT-PLT transfusion. RESULTS: Over the course of 7 years in the study centres, 4067 patients received 19,175 PCT-PLT transfusions. Adverse events were infrequent, and most were of Grade 1 severity. On a per-transfusion basis, 123 (0.6%) were classified an acute transfusion reaction (ATR) defined as an adverse event related to the transfusion. Among these ATRs, the most common were chills (77, 0.4%) and urticaria (41, 0.2%). Fourteen SAEs were reported, of which 2 were attributed to platelet transfusion (<0.1%). No case of transfusion-related acute lung injury, transfusion-associated graft-versus-host disease, transfusion-transmitted infection or death was attributed to the transfusion of PCT-PLT. CONCLUSION: This longitudinal haemovigilance safety programme to monitor PCT-PLT transfusions demonstrated a low rate of ATRs, and a safety profile consistent with that previously reported for conventional platelet components.

Complement activation products in liquid stored plasma and C3a kinetics after transfusion of autologous plasma.

BACKGROUND AND OBJECTIVES: Keeping a small stock of liquid plasma readily available for transfusion is common practise in Sweden. We report data on complement activation markers in plasma components during storage in the liquid state and the kinetics of C3a-(desArg) after transfusion of autologous plasma with high content of C3a-(desArg). MATERIAL AND METHODS: Plasma components were prepared by apheresis or from whole blood. C3 fragments (C3a-(desArg), C3d,g, iC3), and soluble terminal complement complex (sC5b-9) were investigated. C3a-(desArg) kinetics was investigated in regular apheresis donors. RESULTS: Apheresis plasma prepared by membrane centrifugation had significantly higher level of C3a-(desArg), C3d,g and sC5b-9 from day 0 and low iC3, than plasma prepared by other methods. By storage day 7, C3a-(desArg)-levels were above the reference value in 88% of all components. After re-infusion of autologous plasma with high C3a-(desArg) content, there were rapid a(1) and a(2)-distribution followed by a slower b-elimination phase. CONCLUSION: Plasma components prepared by different methods and stored in the liquid phase differ significantly in the amount and timing of complement activation. C3a-(desArg) present in plasma is rapidly eliminated after transfusion. Autologous plasma could be used to study complement kinetics in different clinical situations.

Enhanced growth of Staphylococcus aureus after nitric oxide supplementation during simulated extracorporeal circulation.

BACKGROUND: Several factors contribute to postoperative bacterial infections in cardiac surgery. Long operation times and the use of extracorporeal circulation increase the risk of infection. Nitric oxide has been shown to possess a broad spectrum antimicrobial effect. METHODS: In this study, we investigated the effect of nitric oxide on S. AUREUS growth in whole blood during simulated extracorporeal circulation. RESULTS: S. AUREUS growth increased 6.2-fold after 180 min SECC in the presence of nitric oxide. Leukocyte counts remained unchanged without any differences between the groups. We observed a steady increase in markers of oxidative stress and activity of the innate immune system. Myeloperoxidase levels increased 8-fold, and C3a and terminal complement complex by 2-fold after 180 min. CONCLUSION: S. AUREUS growth is not due to the effect of nitric oxide on the innate immune system but from its effect on the bacteria itself. It has been shown that nitric oxide stimulates the expression of inducible lactate dehydrogenase, specific to S. AUREUS, which improves its resistance to oxidative stress, and may give S. AUREUS a survival advantage resulting in increased growth.

World apheresis registry 2003-2007 data.

OBJECTIVES: Seventy-five centers from many countries have applied for a login code to the WAA apheresis registry. Fifteen centers from 7 countries have been actively entering data at the internet site from 2003 until 2007. We report on data from the registry so far. METHODS: This is a web-based registry. A link is available from the WAA homepage (www.worldapheresis.org). So far data from 2013 patients (12,448 procedures) have been included. A median of 6 treatments have been performed (range 1-140). Mean age 51 years (range 1-94 years; 45% women). Seven percent of the patients were < or = 21 years and 4% were < or = 16 years. RESULTS: The purpose of the apheresis procedure was therapeutic in 67% and retrieval of blood components in 33%. Main indications: neurological and hematological diseases, lipid apheresis and stemcell collection (autologous, and some allogeneic). Blood access: peripheral vessels (71%), central dialysis catheter through jugular (6.5%) or subclavian veins (6.7%), femoral vein (8%) and AV fistula (4%). ACD was used for anticoagulation in 73% of the procedures. Albumin was mainly used as replacement fluid. Adverse events (AE) were registered in 5.7% of the procedures. AE was graded as mild (2.5%), moderate (2.7%) or severe (0.5%). No death occurred due to treatment. The procedures were interrupted in 2.6%. Most frequent AEs were blood access problems (29%), tingling around the mouth (20%), hypotension (18%), and urticaria (9%). There were significant differences between the centers regarding mild and moderate AEs. Data indicate that centers using continuous infusion of calcium had fewer AEs. CONCLUSION: There was a limited number of severe AEs. Centers use various standard procedures for apheresis. By learning from the experience of others the treatment quality will improve further. In the near future, an update of the registry will enable more extensive evaluation of the data.

Ependymoma of the ovary. A clinico-pathologic, ultrastructural and immunohistochemical investigation. A case report.

A case of primary malignant ovarian ependymoma is described. The course of the tumor disease was extremely prolonged with a 50 year history. The diagnosis is supported by immunohistochemical and ultrastructural evidence of ependymal differentiation. The histogenesis and the origin from a possible preexisting ovarian teratoma are discussed.

Steroid production in vitro of a hilus cell tumor of the human ovary.

A 69-year-old heavily virilized woman with an androgen-producing tumor of the right ovary is described. After tumor removal, plasma testosterone levels fell from 5 to less than 0.6 ng/ml. Serum gonadotropins were low prior to surgery and rose to high levels postoperatively. Histologic examination of the right ovary revealed a hilus cell tumor. Incubation of small specimens of tumor tissue for 2 hours in oxygenated Krebs bicarbonate buffer containing glucose and bovine serum albumin yielded a release of predominantly testosterone and androstenedione into the medium. Human chorionic gonadotropin (hCG) added to the medium had no effect on steroid release. Incubation of tumor tissue in vitro may provide a useful functional adjuvant to the morphologic characterization of hormone-producing ovarian tumors. Such combined studies may increase our knowledge of the much-debated histogenesis of these tumors.

Unexpected effects of donor gender on the storage of liquid plasma.

BACKGROUND: Swedish regulations in effect since 2006 allow the storage of plasma for transfusion up to 14 days at 2-6 degrees C and for 3 years at < or = -30 degrees C. In this study, the quality of currently used plasma components was investigated. MATERIALS AND METHODS: Plasma components, prepared from whole blood or by apheresis, either leucocyte depleted or not leucocyte depleted, were stored at 2-6 degrees C as liquid plasma or as thawed fresh-frozen plasma; 31% were from female donors. Concentration, function and activation markers of the plasma coagulation systems were investigated during storage for up to 42 days. RESULTS: Cold-induced contact activation was the dominant storage lesion, occurring earlier and at higher frequency in plasma from females. Increased kallikrein-like activity led to changes in activated partial thromboplastin time, prothrombin time, protein C and C1 inhibitor (C1INH). C1INH function dropped to 53% on Day 14 in cold-activated plasma components. CONCLUSION: Contact activation may be triggered before Day 14, especially in plasma from females, and may progress as a result of the consumption of C1INH. The data suggest that lack of cold-induced contact activation may be an important quality criterion. To achieve this, plasma from male donors could be selected for transfusion and the storage time limited to 7 days.

Renal infection after ileal conduit urinary diversion. An autopsy study.

Autopsy records from cases with urinary bladder carcinoma, consisting of patients operated upon with conduit urinary diversion and controls without diversion, were studied. We found macroscopical evidence of upper urinary tract infection (renal abscesses and/or pus-filled pelves/ureters and/or reddish mucosa in pelves/ureters) in 18 of 21 conduit cases without evidence of cancer at autopsy and in 14 of 50 control cases. The difference is statistically significant (p less than 0.0005). Abscesses were found more often in conduit cases (10/21) than in non-diverted cases (8/50) (p less than 0.01). The known connection between dilated ureters and/or pelves and urinary tract infection was demonstrated in non-diverted cases.

Standardized units of RBCs: is it time for implementation?

BACKGROUND: Current practice for the preparation of RBCs from whole blood for transfusion results in poorly standardized contents of RBC Hb. The principle of apheresis, metering the anticoagulant into the collected blood, which is pumped into an empty container, allows variation in the collected volume according to properties of the donor. STUDY DESIGN AND METHODS: The total Hb mass of each person in a representative group of Swedish blood donors was evaluated by using Hb concentration and blood volume (BV), with the latter calculated from each donor's weight and height. The number of blood units that could be collected without exceeding 13 percent of the BV was estimated at a standardized content of RBC Hb set at 40, 45, and 50 g. RESULTS: With Hb standards of 45 and 50 g per unit of RBCs, it would be possible to collect 1 unit, but not more, from 93 female donors in the study; with 40 g of Hb as the standard, 2 units could be collected from 6 percent of the donors. Using a standard of 40 g of Hb, it would be possible to collect 2 units or more from 95 percent of 121 male donors. The corresponding figures at Hb standards of 45 and 50 g were 81 and 50 percent, respectively, of the male donors. The largest number of units that could be collected would thus be obtained at a 40-g Hb standard. However, the greatest total mass of RBC Hb would have been obtained at 45 g. Even the yield of plasma would reach a maximum at this RBC Hb standard. CONCLUSION: Depending on the donor's Hb and BV, it is possible to collect either 1 or 2 units of RBCs without exceeding 13 percent of any donor's BV, provided the collected volume of blood in each unit is less than the current standard. Such practice would allow better use of the donor population. Two-unit blood collections may reduce donor exposure in transfusions. Applying a standard at 45 g of RBC Hb per unit was found to permit the collection of maximum RBC Hb and plasma in the evaluated population of Scandinavian donors. Perhaps it is time to discuss a change in current rules for the preparation of RBCs for transfusion.

Increased mucosal synthesis of rheumatoid factor (RF) in coeliac disease.

Increased serum levels of RF have been reported in patients with gluten sensitivity. The objective of this study was to investigate the in vivo secretion of different isotypes of RF in the small bowel in coeliac disease. Nineteen patients were investigated by perfusion of a defined jejunal segment, and the jejunal perfusion fluid was analysed for the presence of IgA and IgM anti-Fc (IgG). Five of the patients studied had serum IgA deficiency. Patients with partial/subtotal villous atrophy but no IgA deficiency (n = 7) had a four-fold increase of IgM-RF (P < 0.001) and a three-fold increase of IgA-RF (P < 0.001) compared with healthy controls (n = 29). Patients with normal jejunal mucosa but no IgA deficiency (n = 7) had similar IgA-RF and IgM-RF concentrations to healthy controls. Patients with serum IgA deficiency had no IgA-RF detectable in jejunal fluid but the highest IgM-RF concentrations, in particular in active disease. The coeliac patients had serum levels of IgA-RF and IgM-RF within the reference ranges. Jejunal fluid levels of IgA-RF and IgA-anti-gliadin antibodies were significantly correlated (P < 0.001). The data indicate that enhanced jejunal mucosal production of RF occurs above all in active coeliac disease. The findings suggest that the immune response to gluten induces a mucosal RF synthesis.

HLA antigen sharing in preeclampsia.

The association between human leukocyte antigens (HLA) and the development of preeclampsia (PE) was studied in 48 women with PE and their partners. In 20 of the families, the HLA antigens of the child were also determined. In 8 of 15 cases (53%) of primigravidae with PE, the child shared at least 2 paternal antigens with the mother, whereas this was the case in only 1 of 26 (4%) children of healthy mothers (p < 0.001). In contrast to this, none of 6 children of multigravidae with PE shared 2 paternal antigens with the mother. An increased homozygosity at the HLA-B locus was also seen in the children of primigravidae with PE (40 vs. 4%, p = 0.01).

Pre-separation storage of whole blood: the effect of temperature on red cell 2,3-diphosphoglycerate and myeloperoxidase in plasma.

BACKGROUND: Although whole blood intended for component preparation is commonly left to cool at ambient temperature, knowledge is insufficient concerning what effects this may have on red blood cell (RBC) quality, in particular after a prolonged hold. STUDY DESIGN AND METHODS: Whole blood collected in CPD was incubated at 20 degrees C and 28 degrees C for 6 h designed as a paired study. Blood components were prepared and the red blood cell concentrates (RBCs) were stored for 28 days at 4 degrees C +/- 2 degrees C. Blood gases, pH, glucose, lactate, adenosine triphosphate (ATP), 2,3-diphosphoglycerate (2,3-DPG) and plasma myeloperoxidase (MPO) were investigated. RESULTS: After 6 h the 2,3-DPG concentrations had lowered to 88% (20 degrees C) and 54% (28 degrees C) of initial levels, respectively. The difference was significant and was maintained for 28 days, although, at low levels from day 7 (28 degrees C) and day 14 (20 degrees C) of storage. ATP was maintained at the initial level in both groups during the first 6 h of storage but after component separation the levels were significantly higher in the 28 degrees C group during the first 5 days. The release of myeloperoxidase (MPO) was significantly higher in the non-cooled group than in the cooled group. CONCLUSIONS: Pre-separation holding for 6 h of whole blood at temperatures of 28 degrees C causes a great and rapid loss of 2,3-DPG and considerable formation of acid metabolites resulting in clearly subnormal 2,3-DPG levels even on day 1. Active pre-separation cooling to 20 degrees C is to be recommended.

Increased anticoagulant osmolality improves separation of leukocytes from red blood cells (RBC).

BACKGROUND: The bottom-and-top (BAT) procedure separates the buffy coat (BC) from plasma and red blood cells (RBC). The contents of mononuclear cells (MNC) remaining in the RBC are about 1 x 10(6) cells/unit, whereas the granulocytes are removed less effectively, 500-800 x 10(6) or more remaining in the RBC unit. The aim was to improve the separation efficacy by collecting the blood in an hyperosmolar anticoagulant, followed by BAT separation. It was expected that the red cells would shrink, thereby increasing their density, while the granulocytes would not change volume and density. STUDY DESIGN AND METHODS: 18 donors were included in the study, 12 in the test group and 6 in the control group. CPD-SAGM bags were used, with a modification of the anticoagulant by removal of 20-ml CPD from all units and addition of 20-ml mannitol (test group) or 20 ml of isotonic saline (control group). The collected blood units were cooled on butanediol plates for 2-4 h, then centrifuged and separated into components. The levels of leukocytes in the whole blood, the BC and the RBC were determined by flow cytometry gated for intact CD45+ cells. A number of other tests were performed during 42-day storage. RESULTS: The plasma yield was slightly higher in the test group than in the control group (ns). The contents of leukocytes (CD 45+ intact cells) in the RBC units were 32 +/- 20 x 10(6) in the test group and 573 +/- 241 x 10(6) in the control group. The numbers of MNC were 1.2 +/- 0.6 x 10(6) and 2.6 +/- 1.8 x 10(6), respectively. The RBC 2,3-DPG concentration was slightly better maintained in the test group at day 7 of refrigerated storage (p = 0.0027), but most other tested parameters showed no difference during 42-day storage. It was possible to prepare platelet concentrates with good yield using the pooled-BC method. CONCLUSION: This study indicates that considerable improvement in the BAT procedure can be obtained if the anticoagulant is made hypertonic by the addition of mannitol. This is achieved without altering the already low levels of MNC and keeping the same quality.

Storage of whole blood before separation: the effect of temperature on red cell 2,3 DPG and the accumulation of lactate.

BACKGROUND: Although whole blood intended for component preparation is commonly left to cool at ambient temperature, knowledge is insufficient as to the effects this may have on red cell quality, in particular after a prolonged hold. STUDY DESIGN AND METHODS: Whole blood collected in ACD-A (7% wt/wt) and CPD (12% wt/wt) was incubated at 4, 10, 15, 20, 25, and 30 degrees C for 24 hours. Blood gases, pH, bicarbonate, glucose, lactate, and red cell 2,3 DPG were investigated. RESULTS: When the blood was stored at 30 degrees C, the 2,3 DPG concentration decreased within 4 hours from 858 +/- 106 to 316 +/- 172 mmol per mol of hemoglobin (a 63% decrease); 99 percent was lost within 18 hours. At 25 degrees C, 46 percent was lost within 4 hours and 94 percent within 18 hours; at 20 degrees C, the decrease at 18 hours was 62 percent and that at 15 degrees C was 24 percent. No loss of 2,3 DPG was observed at 4 degrees C and 10 degrees C storage. No difference was attributable to the anticoagulant used. After 24 hours, the lactate concentration at 15 degrees C was 2.9 times the original, that at 20 degrees C was 3.8 times the original, that at 25 degrees C was 7.0 times, and that at 30 degrees C was 9.2 times. CONCLUSIONS: With current anticoagulants, storage of whole blood at temperatures of 25 to 30 degrees C before separation causes a great and rapid loss of 2,3 DPG and an accumulation of acid metabolites. In a hold of blood for >4 hours, rapid cooling is desirable to avoid initial loss of 2,3 DPG.

In vitro production of cyclic AMP and steroids from an ovarian Sertoli-Leydig cell tumor. Notes on clinical management.

A 27 year old nulliparous woman with a history of chronic anovulation and signs of virilization with a markedly elevated serum level of testosterone, underwent a laparotomy with peroperative bilateral ovarian vein catheterization and bilateral bisection of both ovaries. A solid, 1.5 cm, well delimited tumor located centrally in the right ovary, was excised. Testosterone levels in ovarian venous blood from the tumor bearing side, were 88.4 nmol/l and from the contralateral ovary 3.9 nmol/l. Histopathological examination showed a Sertoli-Leydig cell tumor which was radically extirpated. Postoperatively, the serum levels of androgen normalized, the woman had regular cycles, became pregnant and delivered a normal female baby. Pieces of tumor tissue were incubated for 2 h, with and without addition of gonadotropins and adrenocorticotropic hormone (ACTH). Human chorionic gonadotropin (CG), follicle stimulating hormone (FSH) and adrenocorticotropic hormone (ACTH) caused significant increases in cyclic monophosphate (cAMP) production in tumor tissue in vitro, as compared to controls. Furthermore, ACTH also significantly stimulated 17 beta-estradiol production. In tumor cells cultured for 48 h, FSH slightly, but not significantly, increased the production of progesterone. In the cell culture, [3H]-thymidine incorporation into deoxyribonucleic acid (DNA) was stimulated by IGF1 alpha but not by hCG and FSH. It is concluded that Sertoli-Leydig cell tumors may be sensitive to gonadotropins and ACTH and that their small size, solid shape and intra-ovarian localization can cause diagnostic difficulties.