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F Bertolini

Publications and source records attributed to F Bertolini.

At least 73 records · Page 4Linked to original sources
PubMed1994

A multicenter evaluation of reproducibility of swirling in platelet concentrates. Biomedical Excellence for Safer Transfusion (BEST) Working Party of the International Society of Blood Transfusion.

BACKGROUND: Eleven laboratories participated in a study investigating whether swirling can be used in monitoring platelet discoid morphology in platelet concentrates (PCs). STUDY DESIGN AND METHODS: In each laboratory, two readers observed the swirling pattern in 1- to 5-day-old PCs, in PCs containing spherical platelets obtained by 4 degrees C storage (spherical PCs), and in PCs containing known proportions of spherical and discoid platelets. Swirling was reported as positive, intermediate, or negative. Moreover, platelet morphology was evaluated by oil-phase microscopy. RESULTS: Swirling of spherical PCs was found to be positive and negative in 5 and 68 percent of cases, respectively. Conversely, swirling of fresh PCs was positive and negative in 83 and 2 percent of cases, respectively. Results in PCs containing known amounts of added spherical platelets were intermediate between those in fresh and spherical PCs. The observed agreement between the two readers (number of concordant readings/total readings) was 67 percent. Most disagreements concerned positive versus intermediate or intermediate versus negative evaluations, whereas a positive versus negative disagreement was observed in only 1 percent of cases. The percentages of discs found by microscopy showed greater variation among the different laboratories, indicating that this measurement should be better standardized. CONCLUSION: Evaluation of swirling seems promising for large-scale quality control of PCs. Further studies are needed to confirm this hypothesis.

Blood Platelets↗
PubMed1994

Retrovirus-mediated transfer of the multidrug resistance gene into human haemopoietic progenitor cells.

We report the utilization of cord blood (CB) or bone marrow (BM) derived low density or purified CD34+ cells as a target for human multidrug resistance (MDR1) gene transfer. Cells were cocultivated for 48 h with an irradiated MDR1 retroviral producer line. Since some degree of MDR1 gene expression has been reported to occur in haemopoietic progenitor cells and in peripheral blood cells, efficiency of MDR1 gene transfer was assessed by: (1) Drug selection and culture in presence of 50 ng/ml doxorubicin, 10 ng/ml colchicine and 0.85 micrograms/ml taxol. In uninfected control, 1-2% of CFU-GM and CFU-GEMM were found to be drug-resistant, while 14-31% of original clonogenic activity was found after 2 weeks of culture of transduced cells. Efficiency of MDR1 transfer was significantly enhanced by prestimulation with cytokines, and found to be significantly superior in CB-derived compared to BM-derived progenitors. (2) Analysis of MDR1 gene expression by evaluating MDR1 mRNA through polymerase chain reaction. MDR1 expression was very low in cultures of uninfected controls, whereas, after drug selection, MDR1 mRNA levels in transduced cells was as high as in the MDR1 retroviral producer line (positive controls). (3) Flow cytometric analysis of the expression of CD34 and P-glycoprotein, the product of the MDR1 gene. After MDR1 transduction and 2 weeks of culture, membrane expression of P-glycoprotein was found on 17-25% of viable CD34+ cells. (4) Cytochemical localization by APAAP staining of P-glycoprotein. No specific localization was found in untransduced controls, whereas transduced and cultured CB-cells expressed P-glycoprotein on plasma and nuclei membrane. In conclusion, MDR1 gene transfer into CB- and BM-derived progenitor cells seems a feasible and attractive approach to generate a drug-resistant haemopoiesis.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1994

Cord blood plasma-mediated ex vivo expansion of hematopoietic progenitor cells.

Cord blood (CB) plasma has previously been found to augment the replating capacity of CB-derived hematopoietic progenitors. In the present study, we observed a 2.8 +/- 0.3 and a 1.8 +/- 0.2-fold expansion of CFU-GM in cultures of CB cells without growth factors but supplemented by CB plasma or maternal blood (MB) plasma, respectively. In the absence of growth factors, CFU-GM expansion did not occur in CB cell cultures supplemented with fetal calf serum (FCS), peripheral blood (PB) plasma, PB plasma plus concentrations of IL-6 similar to those previously reported in CB plasma and in bone marrow (BM) cell cultures supplemented with FCS, CB, MB or PB plasma. In the presence of stem cell factor (SCF), IL-3 and IL-11, an expansion of CFU-GM was observed in CB and BM cell cultures supplemented by either CB, MB or PB plasma or FCS. Nevertheless, progenitor expansion was significantly superior in CB cell cultures supplemented by CB plasma (8.7 +/- 0.7, 2.4 +/- 0.3 and 2.6 +/- 0.3-fold expansions for CFU-GM, BFU-E and CFU-Mix, respectively). In conclusion, an unknown factor(s) present in CB plasma, probably capable of crossing the placenta, has an effect alone and in the presence of growth factors in supporting ex vivo expansion of CB progenitors.

Adult↗
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↗
PubMed1993

Role of lactate in platelet storage lesion.

It is known that lactate accumulation may cause a pH fall in platelet concentrates (PC) during storage, and this phenomenon causes platelet morphological lesions and loss of platelet in vivo viability. In this study, we added increasing amounts of lactate to identical PC in order to evaluate the role of hydrogen ion accumulation in determining platelet activation and lesion during storage. Six hours after PC preparation, lactate was added to PC1 and PC2 at 20 and 12 mM final concentrations, respectively, while PC3 served as control. In PC1, pH was lower than 6.3, and platelet function and discoid morphology were lost. PC2 were stored for 7 days at pH values ranging from 6.4 to 6.6, and most results of in vitro measurements reflecting platelet function such as osmotic reversal, ATP release and aggregation in response to different stimuli were not significantly inferior when compared to controls. The addition of lactate had no apparent effect on the rise of platelet activation markers P-Selectin, lysosome-like protein gp 53, platelet-bound fibrinogen and granulophysin, while a reduction of borderline significance was observed in glycoprotein Ib expression after pH reduction to values lower than 6.6. It is concluded that the rise of platelet activation markers during storage reflects platelet lesions different from those determined by lactate per se.

Blood Platelets↗
PubMed1993

Cord blood banking for stem cell transplant.

Umbilical cord blood has been recently used as a source of hematopoietic progenitor cells for transplantation of pediatric patients. This study was performed to evaluate the feasibility of a cord blood bank for unrelated transplant. When the umbilical cord was clamped within 20 seconds after delivery, it was possible to collect 86 +/- 25 ml of cord recipients with more than 2000 CFU-GM/kg; 53% of cord blood samples were found to contain enough CFU-GM for engraftment in 50-70 kg adult patients.

Adult↗
PubMed1993

Single-donor platelet concentrates stored in synthetic medium. In vitro and in vivo studies.

Single-donor platelet concentrates (PC) were prepared in 80-120 ml plasma and stored in two polyolefin bags after addition of 250 ml plasmalyte, a simple, glucose-free synthetic medium that was previously used for platelet storage; when compared to PC stored in plasma, PC stored in plasmalyte, showed similar platelet quality, morphology and function after 5 days of storage. In vivo increments observed after transfusion of PC stored for 5 days in plasmalyte were similar to those observed after transfusion of 1-2 day old PC stored in plasma. Moreover, transfusion of 5-day old PC stored in plasmalyte was associated with correction of prolonged bleeding times in all 3 of the 3 patients evaluated. It is concluded that plasmalyte seems to be promising as a medium for single-donor PC storage.

Blood Donors↗
PubMed1992

Platelet quality after 15-day storage of platelet concentrates prepared from buffy coats and stored in a glucose-free crystalloid medium.

It has been reported previously that platelet concentrates (PCs) prepared from buffy coat pools diluted in a simple, glucose-free medium (BC-PCs) are effective in thrombocytopenic patients after 4 to 12 days of storage. Such preparations produce platelet increments similar to those of traditional PCs prepared from platelet-rich plasma (PRP-PCs) stored for 1 to 3 days. The purpose of this study was to obtain a series of in vitro measurements during storage to allow a more detailed characterization of BC-PCs and a more detailed comparison of BC-PCs with PRP-PCs. At the beginning of storage, the level of (alpha granule membrane protein-140 (GMP-140), a marker of platelet activation, was significantly higher on PRP-PC platelets, and BC-PCs were superior in measurements reflecting platelet function, such as osmotic reversal, ATP release, and aggregation with collagen. Compared to PRP-PCs, BC-PCs were superior in the percentage of discs, total ATP, and glycoprotein lb expression by Day 7. This superiority became more striking on Day 11. Overall, 15-day-old BC-PCs compared favorably to 7-day old PRP-PCs: BC-PCs were superior in ATP release and aggregation with collagen, but they were not significantly different for all other measurements reflecting platelet quality and function. Thus, the quality of platelets in BC-PCs was superior on Day 1, and this superiority progressed as storage continued. In addition, the metabolism of BC-PCs was favorable.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood Platelets↗
PubMed1992

Role of acetate during platelet storage in a synthetic medium.

It has previously been shown that buffy coat platelet concentrates (BC-PCs) stored in a medium made up of approximately 70 percent platelet storage medium (Plasma-Lyte A, PL) and 30 percent plasma (BC-PC-P) are effective in vivo after 9 to 12 days of storage. In addition to sodium, potassium, magnesium, and chloride, PL contains 27 mM (27 mmol/L) sodium acetate and 23 mM (23 mmol/L) sodium gluconate. This study investigated the effect of acetate and gluconate on platelet metabolism. Identical BC-PCs were stored at 22 +/- 2 degrees C in PL (BC-PC-P); PL with gluconate but without acetate, termed PL-A (BC-PC-A); or PL with acetate but without gluconate, termed PL-G (BC-PC-G). On Day 1 of storage, no significant differences were seen between the three groups of BC-PCs. In both BC-PC-P and BC-PC-G, pH and bicarbonate were stable at 7.0 +/- 0.03 and 8.4 +/- 0.9 mEq per L throughout 10 days of storage, whereas in BC-PC-A, they fell to 6.7 +/- 0.05 and 5.5 +/- 0.8 mEq per L on Day 5 (p less than 0.01 vs. Day 1) and to 6.1 +/- 0.1 and 1.2 +/- 0.4 mEq per L, respectively, on Day 10. The buffering capacities of 70 percent PL, PL-A, or PL-G and 30 percent plasma were similar in a platelet-free setting when incremental additions of lactic acid were made. The role of acetate was further studied by adding 14C- or 3H-labeled acetate to BC-PC-P.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetates↗
PubMed1992

Platelet concentrates stored in synthetic medium after filtration.

Platelet concentrates prepared from pooled buffy coats (BCPC) were stored in Plasma-Lyte A, a glucose-free synthetic medium, after leukocyte depletion by filtration through Pall PL 50, and compared to paired unfiltered BCPC stored in the same medium. Each pair of BCPC units was prepared from a pool of 10 buffy coats split into two identical units. Platelet and leukocyte counts per unit of BCPC were 2.70 +/- 0.19 x 10(11) and 3.8 +/- 2.8 x 10(6) (filtered BCPC), 2.59 +/- 0.27 x 10(11) and 79 +/- 56 x 10(6) (control BCPC), respectively. Filtration procedures did not affect in vitro parameters of platelet quality and function such as osmotic reversal, ATP release and aggregation in response to collagen and ADP during 15-day storage. A similar decrease of platelet membrane glycoprotein Ib and a similar rise of activation markers GMP-140, gp 53 and platelet-bound fibrinogen were observed during storage of filtered and control BCPC. Our study indicates that storage of BCPC after filtration is feasible and that a reduction in leukocyte content by filtration to mean cell counts of less than five millions per unit has probably no effect on platelet storage lesion.

Adenosine Diphosphate↗
PubMed1992

[Hormonal changes during surgical stress. Comparison of isoflurane anesthesia, neuroleptanalgesia, propofol].

Our purpose is to verify if different techniques of general anaesthesia can modify the patient's hormonal response to surgical stress. For this extent 35 patients, undergoing Cottle's septoplasty, were randomly divided into three groups, treated using different anaesthesiological techniques (forane, NLA and propofol). No variation was noted between the three groups regarding cortisol increments, while the patients, in whom forane was used for anaesthesia maintenance, manifested less significant variations in prolactin levels. This may be indicative of good forane protection for surgical stress.

Adult↗