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P Rebulla

Publications and source records attributed to P Rebulla.

At least 73 records · Page 4Linked to original sources
PubMed1996

Quality assurance of clinical transfusion practice by implementation of the privilege of blood prescription and computerized prospective audit of blood requests.

Guidelines, algorithms and recommendations have been issued in the attempt to ensure appropriateness of transfusion practice, but the results are less than satisfactory, mainly due to the difficulty to turn paper procedures into actual practice. In our hospital we have tried to overcome this difficulty through the implementation of a quality assurance programme which includes giving the privilege of nonurgent blood prescription to a limited number of physicians and a computerized prospective audit of blood requests. The latter is performed through verification of the compliance of blood requests, which are designed to include a patient's laboratory and clinical data, with hospital guidelines for the proper use of blood. In the 12 months since implementation of the computerized prospective audit the transfusion service has evaluated 7884 requests. Of these, 63.4% (n = 4998) were for red blood cells, 21.1% (n = 1664) for platelets and 15.5% (n = 1222) for fresh frozen plasma. The prospective audit showed that 96.8% and 98.1% of requests for red units and platelets were appropriate, respectively. Conversely, approximately 27% of plasma requests did not comply with guidelines, mainly because the evidence of coagulopathy was missing. However, inappropriateness of plasma requests for elective general surgery decreased from 39% at the onset of the programme to 14% in the last trimester considered. Moreover, the evaluation by retrospective audit of the proportion of patients transfused with both red blood cells and plasma in the perioperative period out of those transfused with red blood cells only, as an indicator of unwanted reconstitution of whole blood, showed that this proportion decreased from 47.6% (320/672) in the 12 months before implementation of computerized audit to 37.8% (244/646) in the following 12 months (difference = -9.8%, 95% confidence interval of the difference from -4.5% to -15.1%; P < 0.005 by chi 2 test). Our initial experience, together with the present system, shows that (1) the restriction of nonurgent blood prescription to a group of clinicians more educated in transfusion medicine than average clinicians practicing in a large multispecialty hospital is feasible; (2) prospective audit is a useful tool for assuring the quality of blood requesting.

Algorithms↗
PubMed1996

Practical guidelines for process validation and process control of white cell-reduced blood components: report of the Biomedical Excellence for Safer Transfusion (BEST) Working Party of the International Society of Blood Transfusion (ISBT).

BACKGROUND: The increased use of white (WBC)-reduced blood components has prompted many institutions to develop quality assurance programs directed to such component preparation processes. For consistent preparation of WBC-reduced blood components that meet clinical needs as well as national standards, a program of process validation and control should be instituted. This involves controlling key factors that affect WBC reduction as well as periodic monitoring of the residual cellular content of components. Practical guidelines for the implementation of such a program are provided. STUDY DESIGN AND METHODS: A program involving three phases of monitoring was developed by individuals belonging to an international working party of the International Society of Blood Transfusion. RESULTS: The first phase, process validation, evaluates a minimum of 20 consecutive units (a minimum of 60 units when nonparametric measurements are used) to document the successful local implementation of a new or substantially modified process. Ongoing process control employing Levey-Jennings type control charts is used to demonstrate that the process remains stable over time. Process capability assessment and conformance with standards are evaluated once residual WBCs are counted in a sufficient number of units. This enables a facility to claim with a specified degree of confidence that a stated proportion of WBC-reduced units will meet national standards. Two approaches to determine the number of units that should be selected for counting are presented. The first approach considers units as either acceptable or not acceptable and assumes that the distribution of failed (or nonconforming) units approximates the Poisson distribution. The second approach takes into consideration the observed WBC content of the tested units, with the assumption that the residual WBC content in WBC-reduced components follows a lognormal distribution. A method to assess the lognormal distribution of residual WBCs is presented. Specific tables based on each of these approaches are provided to guide the reader in the design of a program that will verify conformance with any national standard at specific confidence levels. The approach can be generalized to other process control applications. CONCLUSION: Guidelines are presented for process validation, process control, and assessment of conformance in the production of WBC-reduced blood components. Policy makers retain the responsibility to establish, on the basis of the expected use of WBC-reduced components, requirements for the frequency of testing and for the proportion of prepared units that are expected with a stated degree of confidence to meet the standards. Facilities preparing WBC-reduced components can monitor key factors that influence the preparation of WBC-reduced blood, can periodically assess their conformance with the standards, and can intervene to correct adverse changes in the process. This approach can be used to ensure the consistent quality of WBC-reduced blood components.

Blood Component Removal↗
PubMed1996

Optimal conditions for white cell reduction in red cells by filtration at the patient's bedside.

BACKGROUND: A quality control program of white cell (WBC) reduction in red cells at the bedside was implemented, based on postfiltration counting in a Nageotte chamber of the residual WBCs from samples taken from a segment of the transfusion set, after 1-in-10 sample dilution with Türks's solution. During a 1-year quality control program, 5.1 percent of counted units had apparent filtration failures, that is, WBC counts exceeding 5 x 10(6) per unit. The cause(s) for these apparent failures were investigated. STUDY DESIGN AND METHODS: In Study 1, residual WBCs from 150 buffy coat-free red cells filtered through one type of filter at 4 degrees C, 20 to 24 degrees C, or 27 degrees C in 5 to 10 minutes, 50 to 100 minutes, or 100 to 200 minutes were counted as described above. In Study 2, residual WBCs in samples collected from segments of the transfusion set and from the postfiltration bags were counted in parallel by a new, more sensitive counting method. In this method, 5 mL of filtered red cells was diluted with 20 mL of 3-percent paraformaldehyde and centrifuged, the pellet was resuspended to 500 microL with a lysis solution, and the WBCs were counted in a Nageotte chamber. In Study 3, residual WBCs were counted by the 3-percent paraformaldehyde method in samples from postfiltration bags of 1- to 2-day-old buffy coat-rich red cell units filtered through a second type of filter. Filtration was started within 30 minutes of the removal of the unit from the refrigerator, ambient temperature was 20 to 24 degrees C, and the median filtration time was 90 minutes per unit. RESULTS: Study 1: Median WBC counts per unit increased progressively from 51,000 at 4 degrees C to 934,000 at 27 degrees C, with intermediate values at 20 to 24 degrees C. In no unit did the WBC count exceed 5 x 10(6) if filtration at 20 to 24 degrees C was completed within 100 minutes, while counts in excess of 50 x 10(6) were found at 20 to 24 degrees C and at 27 degrees C with filtration times of 100 to 200 minutes, and 50 to 100 minutes, respectively. Study 2: The relation between segment and postfiltration bag WBC counts obtained by the 3-percent paraformaldehyde method was poor, with the latter being almost always lower than the former. Study 3: None of the 120 units filtered through the second type of filter at 20 to 24 degrees C in 50 to 100 minutes contained more than 3.2 x 10(6) WBCs; the median value was 147,000 WBCs per unit. CONCLUSION: On the basis of the results with the 3-percent paraformaldehyde method, which showed the unreliability of segment counts, a new policy was adopted for quality control of bedside WBC reduction, based on controlling the time of and temperature at transfusion. Bedside WBC reduction in 1- to 2-day-old red cells performed with the second type of filter at 20 to 24 degrees C in less than 100 minutes per unit allowed the preparation of units that meet the standard of fewer than 5 x 10(6) WBCs in all tested cases. Bedside WBC reduction with the second type of filter and under the controlled conditions reported seems effective.

Erythrocyte Transfusion↗
PubMed1996

The Milan Cord Blood Bank and the Italian Cord Blood Network.

We describe the activities of the Milan Cord Blood (CB) Bank and of the Italian Cord Blood Network. By October 31, 1995, 763 units were banked in Milan. Of these, 8 units were used to perform 4 related and 4 unrelated transplants in 5 children and 3 adult patients. Early cord clamping after delivery was found to be crucial to increase the volume of CB collected. This procedure does not seem to be detrimental to the newborn. Of various red cell sedimenting agents used to reduce the unit volume and concentrate progenitor cells, 3% gelatin seems to be associated with the best yields. After a preliminary experience of 2 years, the Italian Cord Blood Network (Gruppo Italiano Amplificazione Cellule Emopoietiche, GRACE) was founded in 1995. The initial activities of GRACE are aimed at the development at the national level of CB banking standard operative procedures in agreement with the draft issued by The North American Task Force for The Development of Standards for Hematopoietic Cell Transplantation. Moreover, a wet workshop has been organized to standardize colony-forming unit (CFU) evaluation. The main goal in Milan is the collection of 5000 CB units. Other issues of interest include CB volume reduction, hematopoietic progenitor purification, ex vivo expansion prior to transplantation, and experimental protocols for gene transfer, such as those related to the multidrug resistance (MDR) gene.

Adolescent↗
PubMed1995

Blood transfusion in beta thalassaemia major.

Conventional treatment of beta thalassaemia major is based on regular blood transfusion from early childhood. Maximum effectiveness of transfusion therapy depends on the following. (1) Availability of safe blood. Donation programmes should aim at retaining repeat donors, who carry decreased risk of transmitting blood-borne infections. Donors should be screened with laboratory tests performed to the highest possible standard of quality. Selection of safe donors can be improved by the adoption of questionnaires containing direct questions on risk factors for transfusion transmissible infections. (2) Use of good quality red blood cells, which should be leucodepleted, preferably by filtration, that can be carried out at the bedside. (3) Regular evaluation of blood transfusion indices, including mean level of haemoglobin maintained, annual blood requirement, daily haemoglobin fall, mean transfusion interval, transfusion reaction rate. This can be assisted by the use of a computerized patient record. (4) Maintenance of a permanent record of the patient's blood group genotype (including at least Rh, Kell, Kidd and Duffy systems) and any red cell antibodies that develop. This is mandatory to ensure optimal survival of transfused red cells. (5) Continuous monitoring of transfusion transmissible infections. (6) Vaccination against hepatitis B of all suitable patients. (7) Intensive iron chelation. This should be done by regular subcutaneous administration of desferrioxamine B. Oral chelators, which are currently under laboratory and clinical evaluation, are not yet available for general use.

Blood Donors↗
PubMed1994

Quality control of red cell filtration at the patient's bedside.

BACKGROUND: Concern has been raised about the quality of white cell (WBC) reduction in blood components when it is performed by filtration at the patient's bedside. Thus, the quality of red cell (RBC) filtration performed at the bedside through two new flatbed WBC-reduction filters was evaluated. STUDY DESIGN AND METHODS: In the laboratory, 25 and 10 RBC units suspended in additive solution were stored for 1 to 2 and 14 to 21 days, respectively, and filtered through each filter. At the end of filtration, an automated complete blood count and a manual WBC count (Nageotte chamber) were determined in two samples collected from 1) a segment clamped at 5 and 25 cm below the filter along the line connecting prefiltration and postfiltration bags and 2) the postfiltration bag. In addition, 10 of the 11 nurses of the hematology outpatient clinic administered to hematologic patients 25 RBC units stored for 1 to 2 days through each type of filter. At the end of transfusion, a segment was collected from the transfusion set and a WBC count was performed as described above. No filter priming or rinsing with saline was done. RESULTS: WBC counts obtained after laboratory filtration in the segments were similar to those obtained from the postfiltration bags and from the segments collected at the bedside in all cases, with the exception of 14- to 21-day-old RBCs filtered in the laboratory through one of the filters, which produced slightly higher WBC counts in the segments than were seen in the postfiltration bags. The difference was not significant. In no case was the count in the postfiltration bag higher than that in the segment. Nurse training was easy, and bedside filtration was associated with no untoward effects. CONCLUSION: The RBC filtration at the patient's bedside can be equal in quality to that performed in the laboratory, at least in such clinical settings as hematology and oncology departments in which blood transfusion is common practice, and if simple training is provided to the nursing staff. Under the conditions of this study, it seems that quality control of RBC bedside filtration is feasible and simple.

Blood Transfusion↗
PubMed1994

Dose of desferrioxamine and evolution of HIV-1 infection in thalassaemic patients.

To study the relationship between the dose of desferrioxamine (DFX) and the progression of the HIV-1 disease in thalassaemia major patients (TMP), 64 seropositive TMP patients were studied. Cumulative incidence of CDC stage IV was calculated using a non-parametric life-table method. The association with the mean daily dose of DFX was tested with a Cox proportional hazards model which was also used to adjust for confounding variables. The median of the mean daily dose of DFX over the seropositive period was 40 mg/kg (range 0-65 mg/kg). Age at seroconversion (P < 0.02) and splenectomy (P < 0.03) were found to be associated with the mean daily dose of DFX. 6.5 years after seroconversion, 11% of those who had been prescribed more than 40 mg/kg of DFX daily had entered stage IV versus 35% of those who had been prescribed a lower dose (P < 0.01). When the dose was taken as a continuous variable it was found that the rate of progression was significantly smaller in TMP receiving a higher dose (P < 0.002), even after adjusting for age and splenectomy (P < 0.02). Although it should be noted that these results were obtained in an observational study, possibly biased by a non-random allocation of the DFX dose, we believe that they are striking enough to support the claim that the role of DFX in the progression of HIV disease should be further evaluated.

Acquired Immunodeficiency Syndrome↗
PubMed1994

Multicenter evaluation of methods for counting residual white cells in leukocyte-depleted red blood cells. The Biomedical Excellence for Safer Transfusion (BEST) Working Party of the International Society of Blood Transfusion.

Two wet workshops were conducted involving 20 laboratories to determine optimum methods for counting residual white blood cells (WBC) in leukocyte-depleted red blood cells (RBC). In both studies, calibration samples containing known concentrations of WBC were prepared by diluting 1-day-old EDTA blood in RBC virtually free of WBC. Study No. 1 was aimed at determining the lower limit of detection and at examining advantages and disadvantages of two methods based on flow cytometry (FC) and on the Nageotte chamber (NC), respectively. This study showed that the lower detection limits for FC and NC were 0.1 and 1 WBC/microliters, respectively. Methods based on FC showed less variability at 0.1-1 WBC/microliters but were equal in performance to counting procedures based on the NC above 1 WBC/microliters. We then designed study No. 2 to evaluate three different protocols using the NC. In protocol 1, samples were diluted 1:10 with Plaxan and with Türk's solution; in protocol 2 a mixture of Türk's solution and Zap-oglobin, a reagent used in manual hemoglobin determinations, was used to dilute the samples 1:5; and protocol 3 involved initial 1:10 dilution of a 1-ml sample with Plaxan followed by WBC concentration into the original 1-ml volume by centrifugation. In each participating laboratory, two technologists independently processed the samples and read the results with an NC. Plaxan and Türk's solution gave comparable results. Interobserver variability was not critical to the results. All three NC methods were valid for counting at concentrations of > or = 1 WBC/microliters.(ABSTRACT TRUNCATED AT 250 WORDS)

Calibration↗
PubMed1994

Frozen platelet plates for platelet antibody detection and cross-match.

We performed 2 studies aimed at developing a frozen platelet panel suitable for platelet cross-matching. The stability of the most important platelet membrane glycoproteins and the reactivity of antigens of the human platelet antigen (HPA) and of the human leukocyte antigen (HLA) systems were evaluated with the platelet suspension immunofluorescence test (PSIFT) in a panel of platelets frozen in microplates with 6% dimethylsulfoxide. In study No. 1 we evaluated platelet reaction with a broad-spectrum weak anti-HLA and a potent anti-HPA-1a antiserum and the expression of glycoproteins Ib and IIb/IIIa complex on platelet membrane before freezing and after 0.5, 1, 2, 3, 4, 5, 6 and 12 months of storage at -80 degrees C. In study No. 2 we examined platelet reactivity with anti-HPA-1b, -HPA-2a, -HPA-3a, -HLA-A2, -HLA-A3 of platelets stored frozen for 12 months in parallel with fresh platelets from the same donors. Study No. 1 showed that glycoprotein expression was stable and that the weak anti-HLA and the potent anti-HPA-1a antibodies were clearly detected during 12 months at -80 degrees C. Of the 35 paired PSIFT performed in study No. 2 with fresh and frozen/thawed platelets incubated with anti-HPA-1b, -HPA-2a, -HPA-3a, -HLA-A2, -HLA-A3 antisera and AB serum, concordant reactions were obtained in all cases with the exception of 1 case of HLA-A3-positive platelets incubated with anti-HLA-A3 antiserum, that was reactive with frozen/thawed platelets but nonreactive with fresh platelets from the same donor.(ABSTRACT TRUNCATED AT 250 WORDS)

Antigens, Human Platelet↗
PubMed1993

Platelet concentrates from buffy coats: improved conditions for preparation and evaluation in routine clinical use.

Since 1990, platelet concentrates prepared by soft centrifugation of buffy-coat pools diluted with a glucose-free, commercially available crystalloid solution (BC-PC) are the first choice product for all platelet recipients in our institution. Numerous in vitro and in vivo observations from our own and other laboratories indicate that BC-PC compare favorably to PC prepared from platelet-rich plasma (PRP). In the present in vitro study we evaluated traditional and bottom-and-top bags and modified centrifugation conditions with the aim of increasing in vitro platelet yield in BC-PC. This was 14-18% higher compared with our previous protocol when prolonged centrifugation and bottom-and-top bags were used. In addition, we evaluated post-transfusion platelet count increments in 42 unselected adult hematological patients routinely transfused with 703 1-5 day-old BC-PC pools. Transfusion data were managed with PLATELET, an MS-DOS compatible program which includes automated calculation of transfusion efficacy and periodic patient reports. Mean pre-, 1 h and 24 h post-transfusion platelet counts were 16, 38 and 28x10 9/L, respectively. Mean 1 h and 24 h post-transfusion platelet count increments, expressed as percentage of expected, were 40 and 24%, respectively. These data were similar to those obtained previously in 189 unselected hematological patients given 2432 PRP-PC transfusions (mean 1 h post-transfusion increment 46% of expected). The present in vitro study confirms that similar platelet yields can be obtained with the BC and PRP methods. In vivo findings show that also in routine conditions post-transfusion increments of PRP-PC and BC-PC are similar.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1993

White cell-reduced red cells prepared by filtration: a critical evaluation of current filters and methods for counting residual white cells.

White cell (WBC) reduction, red cell (RBC) recovery, and filtration time were determined in 1-day-old standard and buffy coat-depleted RBCs filtered in the laboratory through six commercial filters for WBC reduction. Residual WBCs were counted with a Bürker chamber (BC), with a Nageotte chamber (NC), and by flow cytometry (FC). Results show that BC counts were 0 in several cases in which WBCs were detected with NC and FC, which indicated that the traditional BC method is too insensitive in use with currently available filters. Calibration curves performed by FC and with NC with samples containing known concentrations of WBCs from 1000 to 1 per microL showed that both FC and NC detected, on average, 67 percent of WBCs present in the samples (efficiency). However, the efficiency of FC showed small variability (61-70%) at different WBC levels, whereas the variability with NC was large (39-91%). This greater variability prevented the correction of NC counts by using a single factor and indicated difficulty in NC standardization. Therefore, because our main aim was to compare different filters rather than to define absolute levels of WBC contamination, uncorrected FC and NC counts were chosen to be reported. True WBC counts per unit should not exceed values that can be obtained by dividing uncorrected counts by the lowest efficiencies (61% for FC and 39% for NC). Uncorrected NC and FC counts were below 2 x 10(6) per unit in all units processed through three of the filters and below 5 x 10(6) per unit in all units processed through the other three.(ABSTRACT TRUNCATED AT 250 WORDS)

Erythrocytes↗