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Immunohistochemical detection of breast cancer cells in paired peripheral blood progenitor cell specimens collected after cytokine or cytokine and myelosuppressive chemotherapy.

Mobilized peripheral blood progenitor cells (PBPC) from 30 patients with advanced breast cancer were studied for the presence of tumor cell contamination using a highly sensitive immunohistochemical technique with the capacity to detect one tumor cell in one million mononuclear cells. Aliquots of PBPC were obtained after 4 days of G-CSF and/or GM-CSF and again during G-CSF-stimulated recovery from myelosuppressive doses of cyclophosphamide. The overall incidence of tumor cell contamination was 23%, occurring in PBPC specimens from seven of 30 patients. All four cases in which tumor cells were detected after mobilization with cytokine alone also had tumor cells detected in PBPCs collected following chemotherapy and G-CSF. There were three cases in which malignant contamination was detected only in the specimens collected after cyclophosphamide. There was a greater frequency of tumor cell contamination in aphereses performed during G-CSF-stimulated recovery from cyclophosphamide than in collections primed by cytokine alone (13% vs 23%; P = 0.08), although this did not reach statistical significance. This trend suggests that collection of PBPC during cytokine-stimulated recovery from myelosuppressive chemotherapy may be associated with a greater risk of contamination with malignant cells than apheresis during mobilization with cytokines in the steady state.

Adult↗

Autologous umbilical cord blood transfusion.

The purpose of this study was to examine some aspects of umbilical cord blood collection for autologous transfusion in premature infants. All 120 microbacterial cultures (aerobic and anaerobic) of cord blood samples as well as 30 cultures of mycoplasma were treated. Cord prothrombin fragment (F 1 + 2) concentrations were quantified at one and 10 minutes after clamping of the cord. F 1 + 2 concentrations assessed on 25 newborn infants were similar and no linear association with time of clamping could be drawn. This means that cord blood thrombosis is not activated for at least 10 minutes following clamping of the cord. As far as is known, the first newborn infant to benefit from this method of transfusion is reported here. The premature infant received two portions of autologous blood (on days 5 and 7). No untoward effects were noted. Blood, collected from the umbilical cord, is a safe source for autotransfusion, provided that bacteriological testing has been carried out.

Blood Coagulation↗

Current performance of patient sample collection in the UK.

Collection of the patient sample for pretransfusion testing begins a complex chain of events in the transfusion process. Hospitals in England and North Wales were surveyed to compare local policies against recommendations of the British Committee for Standards in Haematology (BCSH). Hospitals also measured the frequency of rejected and miscollected samples [designated as wrong blood in tube (WBIT)]. 185 of 360 (51.4%) hospitals returned questionnaires and 182 of 185 (98%) hospitals reported that a policy for sample collection existed. Apart from frequent omission of the gender of the patient, there was 96% compliance with all mandatory identifiers of the BCSH guidelines. Practice allowing additions or changes to labelling on sample tubes and request forms varied. 3.2% (14 114/445 726) of samples submitted were rejected for various reasons, the most frequent being incomplete or missing information (49.5% of the total rejected samples). The corrected mean frequency for WBIT in the 27 hospitals with one or more observed WBIT was 1 in 1501 samples (95% CI: < or =1129.09 to < or =1872.91), and the median corrected frequency for WBIT was 1 in 1303 samples. This study has identified great variation in the policy and practice for sample collection for pretransfusion testing. Regular tracking of the rates of sample rejection and WBIT could be used to identify poor performance in individual hospitals requiring investigation and action.

Blood Specimen Collection↗

The Tecumseh study. XII. Enteric agents in the community, 1976-1981.

Enteric illnesses in residents of Tecumseh, Michigan, were studied from 1976 to 1981. The frequency of illness among adults and children increased each year in the late autumn, usually in November. This peak of illness preceded the main period of appearance of the rotaviruses, which were less regularly associated with another peak of illness. Rotaviruses were identified in 3.8% of all stool specimens collected; in specimens from children under two years of age, the annual rate of rotavirus identification was 10.4%. All rotaviruses were identified during the period from late December to early April. Bacterial pathogens were isolated from 3.3% of stool specimens, with no concentration in any age group or season. However, most enteric illnesses in the community were not associated with recognized pathogens. Testing of blood specimens collected in 1976-1978 by complement fixation confirmed the seasonal pattern of rotavirus activity. Rotavirus infections were documented in all age groups, even in older adults, and were associated with symptomatic illness.

Adenoviruses, Human↗

Restriction fragment length polymorphism and polymerase chain reaction-HLA DQ alpha analysis of casework urine specimens.

DNA was isolated from casework urine samples previously submitted for toxicological analysis. The quality and quantity of DNA isolated was determined by spectrofluorometry and agarose yield gel electrophoresis. Hae III restricted samples were then resolved by analytical agarose gel electrophoresis, transferred to a membrane by Southern blotting and hybridized with a chemiluminescently-labelled (D2S44) probe. The DNA fragment banding patterns were indistinguishable from the DNA banding patterns of blood specimens collected from the same donor. Only 5 of 20 samples yielded banding patterns and the banding intensity relative to background was low. Genomic DNA was also obtained from casework samples by Chelex extraction, amplified by polymerase chain reaction (PCR) and then genotyped for human leucocyte antigen (HLA) DQ alpha. Of 20 specimens, 13 (65%) were typed correctly producing identical results for urine and blood specimens obtained from the same donor. Aging studies of casework samples and normal samples (from a non-drug using population) were also conducted with PCR-HLA DQ alpha analysis. Results of these studies indicate that amplification by PCR was more likely to produce positive results. Based on these findings, we conclude that PCR-initiated analysis is more suitable than RFLP analysis for individualization of urine samples.

DNA↗

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.

Blood Donors↗

Monitoring blood loss with near infrared spectroscopy.

Experimental research has shown correlation between near infrared spectroscopy (NIRS) and blood loss, but these findings have not been validated in man. Ten blood donors were monitored before, during and for 10 min after blood collection (470 ml) with NIRS. A Somanetics INVOS 4100 oximeter monitored regional haemoglobin saturation in the cerebral cortex (cSO(2)-left frontal area) and from the left calf (pSO(2)). A Critikon 2001 Cerebral Redox Model monitored total (tHb), oxygenated (O(2)Hb) and deoxygenated (HHb) haemoglobin from the right calf. The oxygenation index [HbD]=[O(2)Hb]-[HHb] was derived from the data. cSO(2) (P<0.001), pSO(2) (P<0.001) and HbD (P=0.001) decreased during blood collection. Maximum changes occurred 10 minutes after collection for cSO(2), with a mean fall (95% C.I.) of 2.5 (-0.06-4.86)%, at the end of blood collection for pSO(2), with a mean fall (95% C.I.) of 3 (0.74-5.26)% and after 8% of blood volume loss for HbD, with a mean fall (95% C.I.) of 7.2 (2.25-12.16). Cerebral and peripheral oxygenation did not recover after blood collection. There was good correlation between NIRS parameters and blood loss. NIRS is a potentially useful technique for monitoring blood loss in humans. Further research is needed to define its role in clinical practice.

Adult↗

Autologous red cells derived from cord blood: collection, preparation, storage and quality controls with optimal additive storage medium (Sag-mannitol).

To investigate whether packed red cells (PRCs) prepared from autologous cord blood-packed red cells (AC-PRCs) could be used as an alternative for homologous-packed red cells (H-PRCs), we developed a system to collect and prepare AC-PRCs and determined standard storage parameters during 35 days of storage in extended storage medium (Sag-mannitol). We collected and fractionated cord blood from 390 newborns. The amount and quality of the AC-PRCs were analysed. The bacterial contamination rate was 1.84%. Twelve AC-PRCs were stored for 35 days, and standard laboratory parameters were measured at day 1 and day 35. The initial laboratory parameters of the AC-PRCs were similar to the parameters of the H-PRCs. After 35 days, the AC-PRCs displayed an increased haemolysis rate compared to H-PRCs (1.1 versus 0.2%) and also a significant decreased adenosine triphosphate value (1.2 versus 2.3 micromol L(-1)). Haemoglobin, haematocrit and pH were comparable in both groups. AC-PRCs meet the quality criteria for H-PRCs after 35 days. Utilizing a closed collection system for cord blood and an extended storage medium will increase safety and quality and facilitate the routine transfusion of autologous red cells derived from cord blood.

Adenosine Triphosphate↗

The need for routine rubella antibody testing of women.

Results of rubella antibody tests performed by the California State Viral and Rickettsial Disease Laboratory on blood specimens collected in 1968 and 1969 from school children and women of childbearing age showed a slightly lower prevalence of rubella antibody in California than reported from most other areas of the United States. Among women of childbearing age, rubella hemagglutination-inhibition (hi) antibody was found in 72 percent of those tested in California compared with 80 percent to 90 percent in other areas of the country. Rubella antibody testing services offered by the State Virus Laboratory included situations in which a pregnant woman was exposed to a suspected case of rubella. It was shown that very few of these exposures constitute a significant risk to the fetus as most of the women already possessed antibody to rubella from past infection and in many instances the exposures were not to actual cases of rubella. The results of this study emphasized the urgency of obtaining blood specimens from pregnant women as soon as possible after exposure to rubella or development of symptoms of rubella. The urgency and anxiety attending these situations can largely be obviated if routine rubella antibody testing of women is carried out prior to pregnancy or at the first prenatal visit.

Adolescent↗

Preparation and storage characteristics of white cell-reduced high-concentration platelet concentrates collected by anapheresis system for transfusions in utero.

BACKGROUND: Important concerns with regard to in utero platelet transfusions are avoidance of volume overload and the immunomodulatory effects of residual white cells (WBCs). This study evaluated a modification of a leukocyte-reduction system (LRS, Spectra, COBE BCT) for apheresis, which collects high-concentration WBC-reduced platelets (HCPs) for in utero transfusion. STUDY DESIGN AND METHODS: The LRS procedure was modified by running the platelet collection pump at specified low flow rates (Q(col)) for the first part of the procedure, collecting HCPs by gently purging them from the LRS chamber into a designated collection bag and then restoring the original LRS procedure settings to collect a second standard apheresis platelet concentrate (PC). Two centers carried out 32 procedures. Platelet yield, residual WBCs, and in vitro platelet function studies were evaluated. RESULTS: Platelet concentrations in 60 mL of HCPs were predictable according to Q(col) (r(2) = 0.735). HCP yields varied from 0.9 to 3.2 x 10(11), depending on the desired final platelet concentrations in 60 mL, with an overall average of 1. 92 x 10(11) (n = 32). Apheresis PCs had a mean platelet yield of 2.9 x 10(11) (1.3-4.4 x 10(11), n = 20) and 3.9 x 10(11) (2.2-5.8 x 10(11), n = 12) at concentrations of 1.3 x 10(12) per L for single-needle and dual- needle procedures, respectively. Median WBC counts were 5.6 x 10(3) for HCPs and 2.0 x 10(4) for apheresis PCs, with >99 percent expected to be less than 1 x 10(6). HCP in vitro characteristics were equivalent to those of apheresis PCs at 24 hours after collection. In vitro performance declined over storage as a function of HCP yield. HCP pH at 22(o)C was maintained at a level of >6.2 for more than 3 days for yields >1.6 x 10(11), less than 2 days for yields 1.6 to 2.2 x 10(11), and less than 24 hours for yields >2.2 x 10(11). HCPs showed good in vitro characteristics and could be stored for 1 to 3 days, depending on the total number of platelets collected. CONCLUSION: A standard apheresis PC and an HCP requiring no secondary processing can be collected with the Spectra LRS. The platelet concentration may be determined by clinical need. HCPs meet the requirements for components that are transfused in utero.

Blood Platelets↗

An international study of the performance of sample collection from patients.

BACKGROUND AND OBJECTIVES: Collection of a blood sample from the correct patient is the first step in the process of safe transfusion. The aim of this international collaborative study was to assess the frequency of mislabelled and miscollected samples drawn for blood grouping. MATERIALS AND METHODS: Hospitals in 10 countries provided data on sample error rates during a period of at least 3 months, including the last quarter of 2001. Mislabelled samples were defined as those not meeting local criteria for acceptance by the laboratory. Miscollected samples [wrong-blood-in-tube (WBIT)] were defined as samples in which the blood group result differed from the result on file from prior testing. WBIT rates were corrected for the proportion of repeat samples and for undetectable errors occurring as a result of chance collection of blood from the wrong patient with the same ABO group. Participants also completed a questionnaire on current policies regarding sample collection. RESULTS: A total of 71 hospitals completed surveys describing policies related to sample collection. Sixty-two hospitals provided usable data on the frequency of mislabelled and miscollected samples. Mislabelled and miscollected samples were common. Based on results from over 690,000 samples, the median hospital performance resulted in a rate for mislabelling of 1 in every 165 samples (6.1 per 1000; interquartile range 1.2-17 per 1000). The presence of national patient identification systems in Sweden and Finland was associated with rates of miscollected samples that were too low to estimate. Outside these nations, miscollected samples demonstrating WBIT occurred at a median rate of 1 in every 1986 samples (0.5 per 1000; interquartile range <0.3-0.9 per 1000). There was great variation worldwide in the reported frequency of mislabelled samples, probably resulting from variation in policies for sample acceptance. Miscollected samples occurred at a more constant rate. CONCLUSIONS: The rate of mislabelled samples and miscollected samples is 1000-10,000-fold more frequent than the risk of viral infection. Rates of mislabelled samples and WBIT can be tracked as key indicators of performance of an important step in the clinical transfusion process. WBIT episodes represent important 'near-miss' errors. By providing baseline performance data for the collection of patient blood samples, this study may be useful in formulating future national standards of performance for sample collection from patients.

Blood Group Incompatibility↗

Accuracy of two systems for blood glucose monitoring without a meter (Chemstrip/Visidex).

Blood specimens collected from 159 campers were tested using Visidex (VD) (Ames Company, Elkhart, Indiana) and Chemstrip (CS) (Bio-Dynamics, Boehringer Mannheim, Indianapolis, Indiana) compared with glucose analyzer (GA) results. Readers were physicians, two with no prior experience using either strip and two with some experience using CS. Subsequently, 30 clinic patients and 4 staff contributed random specimens that were read by a trained novice as 68 discrete samples and compared with GA results. In both studies readers did not place blood on the strips and were thus blinded as to source and the possible previous reading with the other strip. Correlations of VD and CS with GA for camp and clinic data were calculated. Data were compared for reliability and errors greater than 20% of reference values (GA) were compared for VD and CS. CS correlated better with GA than did VD estimates. Reliability was also significantly greater for CS than for VD readings. Both of these visually read methods provided clinically useful estimates of glycemia.

Autoanalysis↗

Allogeneic blood progenitor cell collection in normal donors after mobilization with filgrastim: the M.D. Anderson Cancer Center experience.

BACKGROUND: Information on the safety and efficacy of allogeneic peripheral blood progenitor cell (PBPC) collection in filgrastim-mobilized normal donors is still limited. STUDY DESIGN AND METHODS: The PBPC donor database from a 42-month period (12/94-5/98) was reviewed for apheresis and clinical data related to PBPC donation. Normal PBPC donors received filgrastim (6 microg/kg subcutaneously every 12 hours) for 3 to 4 days and subsequently underwent daily leukapheresis. The target collection was > or =4 x 10(6)CD34+ cells per kg of recipient's body weight. RESULTS: A total of 350 donors were found to be evaluable. Their median age was 41 years (range, 4-79). Their median preapheresis white cell count was 42.8 x 10(9) per L (range, 18.3-91.6). Of these donors, 17 (5%) had inadequate peripheral venous access. Leukapheresis could not be completed because of apheresis-related adverse events in 2 donors (0.5%). Of the 324 donors evaluable for apheresis yield data, 221 (68%) reached the collection target with one leukapheresis. The median CD34+ cell dose collected (first leukapheresis) was 462 x 10(6) (range, 29-1463). The main adverse events related to filgrastim administration in donors evaluable for toxicity (n = 341) were bone pain (84%), headache (54%), fatigue (31%), and nausea (13%). These events were rated as moderate to severe (grade 2-3) by 171 (50%) of the donors. In 2 donors (0.5%), they prompted the discontinuation of filgrastim administration. CONCLUSION: PBPC apheresis for allogeneic transplantation is safe and well tolerated. It allows the collection of an "acceptable" PBPC dose in most normal donors with one leukapheresis, with minimal need for invasive procedures.

Adolescent↗

In vitro analysis of shed blood from patients undergoing total knee replacement surgery.

One method used to obtain autologous blood includes collection of wound drainage postoperatively. The decision to wash wound drainage before infusion is left to individual institutions. The composition of blood collected from joint spaces has not been adequately evaluated. Wound drainage from total knee replacement was collected into a cardiotomy reservoir, without anticoagulation, for 4 hours after surgery. Coagulation parameters were evaluated on the washed supernatant and unwashed supernatant. The most significant findings were the following: 1. Modified prothrombin time: washed supernatant and unwashed supernatant were substituted for tissue thromboplastin reagent. The unwashed supernatant initiated fibrin formation (mean, 108 seconds), whereas the washed supernatant did not (mean, > 150 seconds, P = .01). 2. Euglobulin lysis times: Mixtures containing 50% normal plasma and 50% washed supernatant or unwashed supernatant were used to determine plasmin activation (unwashed supernatant + normal plasma = 24 minutes; washed supernatant + normal plasma = 106 minutes; P = .03). Lower euglobulin lysis times indicates increased plasmin activity. 3. Fibrin degradation products: Concentrations were significantly elevated in unwashed supernatant (mean 10,240 micrograms/mL) versus washed supernatant (mean 5 micrograms/mL, P = .02). Fibrin degradation products are inhibitors of fibrin formation and platelets. The authors conclude the unwashed supernatant from wound drainage collected after total knee replacement contains activated components of the soluble coagulation and fibrinolytic systems, and these substances can be significantly reduced with washing.

Blood Coagulation Tests↗

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 Specimen Collection↗

The dose of granulocyte-colony-stimulating factor after chemopriming treatment does not influence apheresis yield of progenitor cells: a retrospective study of 91 cases.

BACKGROUND: The optimal dose of post-chemotherapy granulocyte-colony-stimulating factor (G-CSF) administration before peripheral blood progenitor cell (PBPC) collection has not been determined as yet, although 5 microg per kg per day has been recommended as the standard dose. This study retrospectively analyzed the effect of G-CSF dose on peripheral blood CD34+ cell collection from 91 patients with hematologic malignancies. STUDY DESIGN AND METHODS: Various doses of G-CSF were administered after several chemotherapeutic PBPC mobilization regimens. According to the dose of G-CSF administered, patients were assigned to two groups. Group 1 included 46 patients who received a low dose of G-CSF (median, 3.6 [range, 2.8-4.6] microg/kg/day). Group 2 included 45 patients who received a standard G-CSF dose of 6.0 (5.5-8. 1) microg per kg per day. Patients in the two groups were matched for age, diagnosis, previous therapy, and chemotherapeutic PBPC mobilization regimens. RESULTS: No difference was observed in the median number of CD34+ cells harvested from each group. The number of leukapheresis procedures necessary to obtain a minimum of 3 x 10(6) CD34+ cells per kg was the same in both groups, and the percentage of patients who failed to achieve adequate PBPC collections was similar in the two groups. CONCLUSION: The administration of low-dose G-CSF after chemotherapy appears equivalent to administration of the standard dose in achieving satisfactory PBPC collection. This approach could allow significant savings in medical cost. A randomized and prospective study is necessary, however, to assess the validity of these conclusions.

Adult↗

Recommendations of the task force on codes and machine-readable symbols.

The task force recommends the use of common, numeric codes for identifying blood, blood components, blood type, collection center and similar information. The task force further recommends the use of the CODABAR and/or the OCR-B symbol for representing these codes in machine-readable form. Blood product codes, label formats, symbol format control characters and a check digit system are recommended.

Blood Banks↗

Transfusions in exotic species.

There has been a considerable increase in the popularity of exotic species as pets, particularly, birds and domestic ferrets. Blood transfusions in birds and ferrets are discussed. There is a brief review of hematology, blood collection techniques, the selection and maintenance of donor animals, and the mechanics of transfusion. Because of the marked differences between birds and mammals, attention is given to avian anatomy and physiology as it applies clinically to blood transfusions.

Anemia↗