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Composition of peripheral blood progenitor cell components collected from healthy donors.

BACKGROUND: Peripheral blood progenitor cell (PBPC) components are being collected from healthy donors for allogeneic transplantation, but the quantity, quality, composition, and variability of PBPCs collected from healthy people given granulocyte-colony-stimulating factor (G-CSF) have not been evaluated. STUDY DESIGN AND METHODS: PBPC components were collected from 150 healthy people who were given G-CSF (5, 7.5, or 10 microg/kg/day) for 5 days. The components were evaluated for white cell (WBC), mononuclear cell, CD34+ cell, neutrophil, platelet, and red cell (RBC) composition. RESULTS: The quantities collected were: WBCs, 35.0 +/- 16.4 x 10(9) (range, 11.9-163.3 x 10(9)); mononuclear cells, 33.3 +/- 14.4 x 10(9) (range, 11.9-139.6 x 10(9)); CD34+ cells, 412 +/- 287 x 10(6) (range, 70-1658 x 10(6)); neutrophils, 1.71 +/- 3.59 x 10(9) (range, 0-27.6 x 10(9)); RBCs, 7.2 +/- 4.0 mL (range, 0-22.1 mL); and platelets, 480 +/- 110 x 10(9) (range, 250-920 x 10(9)). PBPC components collected from people given G-CSF at 7.5 or 10 microg per kg per day contained significantly more CD34+ cells (respectively, 428 +/- 300 x 10(6); range, 70-1658 x 10(6) and 452 +/- 294 x 10(6); range, 78-1380 x 10(6)) than those from people given G-CSF at 5 microg per kg per day (276 +/- 186 x 10(6); range, 91-767 x 10(6)) (p = 0.007 and p = 0.002). When 10 microg per kg per day of G-CSF was given, 50 percent of the components contained enough CD34+ cells for transplantation to a 75-kg recipient (375 x 10(6) CD34+ cells), but 10.6 percent of the components contained less than 150 x 10(6) CD34+ cells and thus would provide a transplantable dose only for a 30-kg patient. CONCLUSION: One PBPC component collected from a healthy donor given 7.5 or 10 microg per kg per day of G-CSF should contain 70 to 1660 x 10(6) CD34+ cells, with 0 to 22 mL of RBCs. Because of the great variability in the number of CD34+ cells collected, the quantity of CD34+ cells in each component should be measured after each procedure to ensure that sufficient quantities of cells are present for a successful transplant.

Antigens, CD34↗

Lipolytic enzyme and phospholipid level changes in intraoperative salvaged blood.

Autotransfusion is becoming increasingly popular, mainly because it eliminates the risk of disease transmission. One of the techniques available is intra-operative blood salvage and retransfusion with or without washing of the collected blood. The blood collected during this process is subjected to a variety of chemical and physical insults which can alter the normal composition of the plasma by activating plasma and cellular homeostatic mechanisms. In this study, we measured the plasma levels of total phospholipids, lysolecithin and non-esterified fatty acids, and the lipolytic enzymes phospholipase A2 (PLA2) and lipase in the salvaged blood before and after washing. In the unwashed salvaged blood the mean levels of PLA2, non-esterified fatty acids and lysophospholipids increased by 144, 96 and 149%, respectively, while those of total phospholipids and lipase did not change to any extent. All these substances were reduced to well below the patients circulating plasma levels by washing the collected blood. The changes indicate that the lipid profile of salvaged blood is significantly altered and that potentially dangerous substances such as PLA2 and its metabolites, lysolecithin and non-esterified fatty acids, are present in increased amounts. Washing the blood is recommended prior to reinfusion.

Blood Loss, Surgical↗

Autologous blood transfusion.

Autologous blood transfusion is a procedure in which blood is removed from a donor and returned to his circulation at some later time. Autologous transfusion can be performed in three ways: (1) preoperative blood collection, storage, and retransfusion during surgery; (2) immediate preoperative phlebotomy with subsequent artificial hemodilution and later return of the phlebotomized blood; and (3) intraoperative blood salvage and retransfusion. All three methods of autologous transfusion offer a potentially superior method of blood transfusion which eliminates many of the problems and complications associated with the banking and administration of homologous donor blood.

Blood Preservation↗

Cardiovascular, haematological and biochemical responses after large volume blood collection in horses.

To determine whether removal of 20 mL/kg of blood (approximately 25% of blood volume) resulted in adverse physiological effects in donor horses, we removed this volume of blood from five horses and selected cardiovascular, haematological and biochemical variables measured during collection and for 31 days thereafter. We found that alteration in most variables occurred, although the changes in values usually remained within published reference ranges. Also, recovery of these alterations to pre-collection values was rapid, occurring within 24--48 h in most instances. We concluded that volumes of blood less than or equal to 20 mL/kg when collected appropriately from healthy donor horses result in no adverse acute or chronic physiological changes. These results suggest that horses undergo adequate physiological compensation when approximately 25% of blood volume is removed for the purposes of blood donation or production of plasma.

Acid-Base Equilibrium↗

Autologous placental blood transfusion for the therapy of anaemic neonates.

Almost 65% of all premature neonates with a birth weight <1,500 g receive at least one erythrocyte transfusion during their first weeks of life. In the present study, we examined the feasibility of autologous transfusions in neonates, using placental blood. Placental blood was obtained from 131 of 141 preterm and term infants using a special placental blood collecting system. Approximately 20 ml of placental blood per kilogram body weight could be harvested, irrespective of birth weight. One placental blood sample was contaminated with maternal erythrocytes; aerobe or anaerobe contamination was observed in any of the stored placental blood products (n = 119) after 35 days of storage. 19 of the 141 newborns needed allogeneic erythrocyte transfusions during the first 12 weeks of life. In 5 of these 19 patients, the amount of placental blood collected would have been enough to dispense with further allogeneic blood transfusions. After completion of the preclinical study, we transfused a total of 22 children, using autologous placental blood. 8 of the 10 infants with a birth weight between 1,000 and 2,000 g and 3 of 5 infants requiring surgical intervention directly after birth needed no further allogeneic blood transfusions. We, therefore, conclude that the collection and preparation of placental blood is feasible for clinical use. The target groups of neonates who are most likely to benefit are infants with a birth weight between 1,000 and 2,000 g and neonates requiring surgical intervention directly after birth.

Anemia↗

Effects of high platelet concentration in collecting and freezing dry platelets concentrates.

OBJECTIVE: To evaluate, in vitro, the effects of collecting and cryopreserving fresh dry platelet concentrates (PCs). MATERIAL AND METHODS: Standard and dry PCs were collected in the same apheresis procedure. PCs were evaluated by mean platelet volume (MPV), pH, glucose and LDH levels. Activation was examined by flow cytometry using anti-CD41, anti-CD42 and anti-CD62p monoclonal antibodies and annexin binding assay. Platelet function was assessed by aggregation using ADP, collagen and arachidonic acid as agonists. Dry PCs were compared to standard PCs and to cryopreserved dry PCs. We also compared the use of ThromboSol to 5% DMSO as cryoprotectives. RESULTS: Dry PCs presented a significantly reduced pH and glucose (p<0.001), increased LDH levels and CD62p expression (p<0.001) and diminished aggregation response to ADP (p<0.001). Platelet cryopreservation was associated with platelet lysis, activation and loss of function. Dry PCs cryopreserved with TS were associated with statistically higher LDH levels (p<0.001) and a higher percentage of annexin binding (p=0.005), in addition to a lower number of CD42 positive platelets (p=0.01). CONCLUSION: Dry PCs should be rapidly frozen after collection to avoid a fall in pH and platelet activation. 5% DMSO performed better than TS to cryopreserve dry PCs.

Blood Platelets↗

Clinical application of quantitative analysis for detection of hematogenous spread of hepatocellular carcinoma by real-time PCR.

In order to detect a hematogenous spread of tumor cells in patients with hepatocellular carcinoma, reverse transcription-polymerase chain reaction assay has been used. In this study, we quantified alpha-fetoprotein (AFP) messenger RNA by real-time PCR approach using LightCyclertrade mark technique. AFP messenger RNA in the blood from 23 hepatocellular carcinoma patients undergoing hepatic resection, 31 healthy volunteers, 10 patients with liver cirrhosis and 5 patients underwent hepatectomy except for hepatocellular carcinoma was quantitated. In the real-time PCR, fluorescence was undetectable in any of the controls. On the contrary, fluorescent signals were detected in 10 out of 39 blood specimens collected from 23 HCC patients. AFP-positive status was significantly associated with the existence of multiple intrahepatic nodules. Out of 8 cases with AFP-positive status, intra- and/or extra-hepatic recurrence has been observed in 3 cases. The quantities of AFP messenger RNA in these 3 cases were relatively high among 8 cases with AFP-positive status. AFP messenger RNA was detectable by newly developed real-time PCR approach with LightCycler and it is suggested that this approach could be applicable in detection of small amounts of tumor cells in the blood of HCC patients.

Biomarkers, Tumor↗

The impact of intrapartum factors on umbilical cord blood stem cell banking.

AIMS: Umbilical cord blood can be used as an alternative source for related and unrelated allogeneic stem cell transplantation. This study was undertaken to determine whether intrapartum factors have an influence on the hematopoietic cell compartment of cord blood. METHODS: Cord blood samples were obtained from 102 normal full-term deliveries for the banking of stem cells. We analyzed the influence of intrapartum factors on the count of CD34+ cells, total nucleated cells, colony forming units and total volume of collection. Fluorescence-activated cell sorting was used to measure CD34+ cell numbers. Statistical analysis was undertaken using Pearson correlation test and multiple regression analysis. RESULTS: The higher the infants' birthweight the larger was the volume. A lower arterial umbilical pH and a larger blood volume resulted in an increased number of CD34+ cells. A large blood volume, long duration of labor, lower arterial and venous pH were correlated with more nucleated cells. A higher birthweight, larger blood volume and lower arterial pH resulted in an increased number of colony forming units. CONCLUSIONS: Some intrapartum factors have an impact on the characteristics of collected cord blood cells. Stress during delivery may influence the number of hematopoetic cells, through altered cytokine production. This knowledge may facilitate the selection of optimal cord blood samples for unrelated banking and the early discarding of suboptimal cord blood samples thus resulting in the saving of costs related to expensive further processing.

Antigens, CD34↗

[Acquired immunodeficiency syndrome. Measures to be taken in blood transfusion establishments in order to protect the personnel].

There is presently no evidence of AIDS transmission to laboratory workers. Because transmissible agent is suspected, precautions should be taken in laboratories. As a general rule, procedures appropriate for hepatitis B prevention are advised. These procedures concern blood collection, carrying, and processing. Measures should be taken in order to avoid blood spilling and work surfaces should be decontaminated. Disposable material should be preferred, and all potentially contaminated material should be decontaminated before disposal or reprocessing. All personnel should be aware of usual laboratory procedures concerning pipetting, hand washing, protective clothing and of general precautions against viral contaminations.

Acquired Immunodeficiency Syndrome↗

Collection of fetal stem cells and newborn effects.

The aim of this work was to test fetal stem cells (FSC) number modification in relation to clamping time and newborn effect. The results show that a fast sample, between 20 and 40 seconds, from umbilical cord after fetus birth and before placental detachment assured a greater quantity of blood useful for the transplants; and that it was necessary to enrich the collected blood in CD34+ cells with specific clonogenic culture, as this is otherwise a small number for a donation to an adult. In the "new donors" the effects of the unconscious donation always depend on the clamping time, which should be the shortest possible to avoid blood overload, which is very dangerous in the presence of heart malformation.

Antigens, CD34↗

Approaches to plasma fractionation for improved recovery and the development of potentially useful clinical factors.

World-wide use of large amounts of highly purified Factor VIII and Factor IX concentrates has increased steadily over the last decade. The increase has resulted in more effective treatment of patients with haemophilia, especially those patients on home care and 'demand' therapy, those on prophylactic therapy for chronic bleeding or those on maintenance therapy for the suppression of inhibitors. Changing socioeconomic conditions have also contributed to greater use of the concentrates. The trend is putting an increasing strain on conventional methods for collecting and fractionating plasma. These include the donor, the nature of the anticoagulant, methods for purification of the clotting factors, procedures for denaturation and/or separation of viral contaminants and patterns of clinical usage. As a result, it is essential that we upgrade the quantity and quality of plasma collected and utilize the most effective new methods for increasing the yield and purity of the plasma fractions. Of special significance are recently described procedures for heat inactivation and/or separation of viral contaminants from the fractions and the long-term promise of genetically engineered proteins.

Antibody Formation↗

Diagnosis of hydatidiform mole and persistent trophoblastic disease: diagnostic accuracy of total human chorionic gonadotropin (hCG), free hCG {alpha}- and {beta}-subunits, and their ratios.

OBJECTIVE: Human chorionic gonadotropin (hCG) is widely used in the management of hydatidiform mole and persistent trophoblastic disease (PTD). Predicting PTD after molar pregnancy might be beneficial since prophylactic chemotherapy reduces the incidence of PTD. DESIGN: A retrospective study based on blood specimens collected in the Dutch Registry for Hydatidiform Moles. A group of 165 patients with complete moles (of which 43 had PTD) and 39 patients with partial moles (of which 7 had PTD) were compared with 27 pregnant women with uneventful pregnancy. METHODS: Serum samples from patients with hydatidiform mole with or without PTD were assayed using specific (radio) immunoassays for free alpha-subunit (hCGalpha), free beta-subunit (hCGbeta) and 'total' hCG (hCG + hCGbeta). In addition, we calculated the ratios hCGalpha/hCG + hCGbeta, hCGbeta/hCG + hCGbeta, and hCGalpha/hCGbeta. Specificity and sensitivity were calculated and paired in receiver-operating characteristic (ROC) curve analysis, resulting in areas under the curves (AUCs). RESULTS: hCGbeta, hCGbeta/hCG + hCGbeta and hCGalpha/hCGbeta show AUCs ranging between 0.922 and 0.999 and, therefore, are excellent diagnostic tests to distinguish complete and partial moles from normal pregnancy. To distinguish partial from complete moles the analytes hCGbeta, hCG + hCGbeta and the ratio hCGalpha/hCGbeta have AUCs between 0.7 and 0.8. Although hCGalpha, hCGbeta and hCG + hCGbeta concentrations are significantly elevated in patients who will develop PTD compared with patients with spontaneous regression after evacuation of their moles, in predicting PTD, these analytes and parameters have AUCs <0.7. CONCLUSIONS: Distinction between hydatidiform mole and normal pregnancy is best shown by a single blood specimen with hCGbeta, but hCGbeta/hCG + hCGbeta and hCGalpha/hCGbeta are also excellent diagnostic parameters. To predict PTD, hCGalpha, hCGbeta, hCG + hCGbeta and hCGalpha/hCGbeta are moderately accurate tests, although they are not accurate enough to justify prophylactic chemotherapy treatment for prevention of PTD.

Adolescent↗

Collection of lymphocytes on a continuous-flow blood cell separator.

Lymphocytes have been collected by continuous flow leukapheresis from cancer patients (pre-treatment) and stored at -190 degrees C in liquid nitrogen as immunological support for intensive chemotherapy. Described here are methods for obtaining maximum yield and viability. Assessments are made of cell count, both during and after leukapheresis, and of viability just prior to storage.

Blood Specimen Collection↗

Cryopreserved red blood cells for pediatric transfusion. Frozen storage of small aliquots in polyvinyl chloride (PVC) plastic bags.

Human nonrejuvenated and rejuvenated red bood cells were prepared for cryopreservation and subsequent pediatric transfusion. Glycerol was added to the red blood cells in the primary polyvinyl chloride plastic collection bag to achieve a concentration of 40 per cent W/V. The red blood cells were concentrated by centrifugation, and the supernatant glycerol was discarded. Each glycerolized unit was divided into four equal aliquots in the individual 600-ml bags of a dry quadruple polyvinyl chloride plastic system, and each aliquot was frozen and stored at -80 C. After thawing, sodium chloride solutions were used to wash the aliquots in the IBM Blood Processor 2991-1 or 2991-2 or the Haemonetics Blood Processor 115, and the washed aliquots were stored in a sodium chloride-glucose-phosphate solution at 4 C for 24 hours. Freeze-thaw recovery of the red blood cells was about 97 per cent, and freeze-thaw-wash recovery was about 84 per cent. Twenty-four-hour posttransfusion survival values were about 92 per cent for both nonrejuvenated and indated-rejuvenated red blood cells. Nonrejuvenated red blood cells, those frozen within three to five days of collection without biochemical modification, had normal oxygen transport function at the time of transfusion; rejuvenated red blood cells, those biochemically treated with PIGPA Solution A after three to five days of storage at 4 C, had improved oxygen transport function at the time of transfusion.

Adenosine Triphosphate↗

The changing relationships in transfusion medicine.

Maintaining quality in provision of transfusion services in the face of mergers, acquisitions, affiliations, and risk-sharing relationships between organizations that formerly conducted business in a traditional vendor-purchaser model is the ultimate challenge. Publications, both lay and professional, highlight the speed and nature of the impetus for change, especially in the United States, where managed care philosophies are driving a bottom-line mentality. Blood collection and transfusion organizations are developing new relationships, including entry of for-profit entities into a formerly virtually exclusively not-for-profit environment, provision of transfusion services by formerly exclusive blood collection entities and vice versa, outsourcing of selected portions, and other innovative relationships, with significantly more competitive marketing strategies. Measures of quality of transfusion services should benchmark current practices, if possible, before entering into new relationships to ensure that the quality of patient care remains high. Concerns about the fiscal viability of organizations should not minimize safety and availability of blood for transfusion when needed.

Blood Donors↗

Isolation of genital mycoplasmas from blood of febrile obstetrical-gynecologic patients and neonates.

In a prospective study, 1156 blood specimens collected from hospitalized febrile obstetrical-gynecologic patients and neonates with suspected sepsis, were inoculated into a conventional biphasic culture medium, Castaneda S and cultures incubated aerobically. 15-24 h later the broth cultures were subcultured to specific media for detection of mycoplasmas. Genital mycoplasmas were isolated in 15 samples (taken from 8 women) and in 2 from 1 neonate. Mycoplasmas and members of the family Enterobacteriaceae were the most frequent significant bacteria isolated from adult specimens. Mycoplasma isolations were associated with either postpartum or postabortum febrile infections in women. Four of the neonates, whose mothers were infected, showed respiratory distress at birth; 1 of them had mycoplasmas in the blood. All febrile states in obstetrical or gynecological patients, and in neonates, should routinely lead to blood cultures for detection of mycoplasmas and ureaplasmas.

Adult↗

Hematology and immunology studies: the second manned Skylab mission.

The hematologic and immunologic functions of the Skylab 3 (second manned mission) astronauts were examined during the preflight, inflight, and postflight phases of the 59-d mission in order to evaluate the response to and/or the influence of the space flight environment. Most changes observed were subtle and did not represent a threat to the health and safety of the crewmen during orbital flight. Even the most significant change observed, a reduction in the circulating red cell mass, did not have a detrimental influence on the astronaut cardiovascular or exercise responses as evaluated by other experiment protocols. Considering the facts that the data were not collected under ideally controlled conditions and that the astronauts were in excellent physical condition, the results of these studies would seem to indicate that man can function quite well in the space flight environment of the Skylab orbiting workshop for extended periods of time.

Adult↗

Feline transfusion practice in South Africa: current status and practical solutions.

Blood transfusion therapy is often under-utilised in feline practice in South Africa. However, it is a technique that can be safely and effectively introduced in practice. Cats have naturally occurring allo-antibodies against the blood type that they lack, which makes blood typing, or alternatively cross-matching, essential before transfusions. Feline blood donors must be carefully selected, be disease free and should be sedated before blood collection. The preferred anticoagulant for feline blood collection is citrate-phosphate-dextrose-adenine. Blood can either be administered intravenously or into the medullary cavity, with the transfusion rate depending on the cat's hydration status and cardiac function. Transfusion reactions can be immediate or delayed and they are classified as immunological or non-immunological. Indications, methods and techniques to do feline blood transfusions in a safe and economical way are highlighted.

Anemia↗