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Biomedical subjects

P Rebulla

Publications and source records attributed to P Rebulla.

At least 109 records · Page 6Linked to original sources

Evaluation of platelet concentrates prepared from buffy coats and stored in a glucose-free crystalloid medium.

Comparison was made between platelet concentrates prepared from pools of buffy coats removed from standard blood donations and stored in a glucose-free, commercially available crystalloid solution (BC-PCs) and standard platelet concentrates prepared from platelet-rich plasma (PRP-PCs). Platelet yield in BC-PCs and PRP-PCs was 59 and 75 percent of donated platelets, respectively. The number of total white cells in 1 BC-PC unit, prepared from a pool of 7 buffy coats, was 21 x 10(6), i.e., 50 times lower than that of 7 units of PRP-PCs. The in vitro values of adequate platelet quality were maintained for 10 days in BC-PCs stored in 1000-mL polyolefin bags. Prolonged bleeding times were reduced or corrected in three of three thrombocytopenic leukemic patients evaluated before and after transfusion of stored BC-PCs. Pretransfusion and 1- and 24-hour posttransfusion median platelet counts in 57 leukemic recipients during 4 months of routine transfusion of BC-PCs (n = 93) were 14, 35, and 27 x 10(9) per L, while those of PRP-PCs (n = 246) were 13, 37, and 31 x 10(9) per L, respectively. No reactions to BC-PCs were reported, but a 1.3 percent rate of reaction to PRP-PC transfusions was reported. This study indicates that BC-PCs are a good alternative to PRP-PCs for platelet support of thrombocytopenic patients.

Adult↗

Correction of the bleeding time in treated patients with severe von Willebrand disease is not solely dependent on the normal multimeric structure of plasma von Willebrand factor.

Even though it is generally held that cryoprecipitate and fraction I-O correct the prolonged bleeding time (BT) in patients with von Willebrand disease (VWD), perusal of reported data indicates that the correction is usually short-lasting and often partial. We decided to do a controlled study of the relationship between the multimeric structure of von Willebrand factor (VWF) and the BT in five patients with severe (type III) VWD after infusion of three plasma concentrates ("wet" cryoprecipitate, lyophilized cryoprecipitate, and fraction I-O) given in random order. The dosage of concentrates was tailored from in vitro measurements to achieve post-infusion levels of ristocetin cofactor above the lower normal limit (50 U/dL) for at least 3 hours. The postinfusion BT became transiently normal in only two of five patients treated with wet cryoprecipitate, whereas it remained prolonged in all five patients treated with lyophilized cryoprecipitate or fraction I-O. For all the concentrates, the proportion of large VWF multimers calculated by scanning the electrophoretic gels were the same as those for normal standard plasmas. An intact multimeric structure was recovered in postinfusion plasmas of patients treated with wet cryoprecipitate, whereas there was a postinfusion loss of large multimers after lyophilized cryoprecipitate and fraction I-O. These findings indicate that the attainment of a normal BT is the exception rather than the rule after the infusion of three plasma fractions used in the treatment of severe VWD, and that an intact multimeric structure in concentrates and postinfusion plasmas is necessary but not sufficient to sustain a normal BT.

Bleeding Time↗

The clinical importance of leukocyte depletion in regular erythrocyte transfusions.

Antilymphocyte, antigranulocyte and antiplatelet alloantibodies, T lymphocyte subsets, expression of HLA-DR antigens on T lymphocytes and NK cell function were determined in 11 homozygous beta-thalassemic children multitransfused ab initio with Erypur-filtered leukocyte-free red cell units (group A) and in 13 similar children multitransfused with standard packed red cell units (group B). No antibodies were found in group A patients, whereas 69% of group B patients were immunized. The two groups did not differ significantly with regard to the other test results. Considered together, thalassemia patients showed a percentage of T4+ cells and a NK cell function that were significantly lower than those found in a reference group of 16 healthy male blood donors. Thalassemics moreover showed a higher than normal percentage of T3+, T4+ and T8+ cells expressing HLA-DR antigens. The results indicate that leukocyte-free red cells should be the treatment of choice for prospective recipients of multiple transfusions, since they are capable of preventing (or delaying) the production of alloantibodies against leukocytes and platelets. From the data of the present study, it does not seem that the transfusion of leukocyte-free red cells is capable of preventing the abnormalities of some immunological tests that occur in some multitransfused patients. Further investigations, however, are needed to draw conclusions on this problem.

Adult↗

Autologous blood predeposit for elective surgery: a program for better use and conservation of blood.

We report the results of a pilot study on predeposit autotransfusion for elective surgery. In 2 years 319 U, each consisting of 350 ml of blood, were donated by 206 patients. Each patient gave 1, 2, or 3 U according to hematocrit levels within 10 days of undergoing surgery on the gastrointestinal tract, lung, liver, kidney, brain, thyroid, arteries, or breast. A hemoglobin drop of about 0.8 gm/dl occurred after each blood donation. A total of 259 autologous and 128 homologous U were transfused to 166 patients during surgery or within postoperative day 4; 40 patients required no blood transfusion. About 70% of patients did not need homologous blood products in addition to autologous units. The most active team of surgeons and anesthesiologists covered about 40% of the patients' blood needs during the study period with autologous units. No patient experienced untoward reactions before, during, or after surgery. Surgeons' and anesthesiologists' responses to the program improved during the study as soon as the advantages of the procedure became clearer. However, gentle pressure from the Transfusion Center was essential for the program's expansion. The patients' acceptance was excellent. We conclude that predeposit autotransfusion is a safe and feasible procedure for transfusion treatment in elective surgery.

Adult↗

Preparation of leukocyte-free platelets for transfusion by filtration through cotton wool.

Filtration through Imugard filters of random platelet concentrates or platelets obtained by plateletpheresis allow the preparation of leukocyte-free platelets for transfusion. The procedure is simple and determines only a small platelet loss (less than 10%). Filtered platelets seem to function normally in vivo. The use of leukocyte-free red cell and platelet transfusions for the support of patients suffering from leukemia or aplastic anemia could prevent major complications, such as refractoriness to platelet transfusion and to bone marrow transplantation.

Blood Platelets↗

Effectiveness of red blood cells filtered through cotton wool to prevent antileukocyte antibody production in multitransfused patients.

The effectiveness of red blood cells made leukocyte-free by filtration through cotton wool to prevent the production of antileukocyte antibodies was evaluated in children suffering from Cooley's anemia. Two studies were performed: study I was carried out prospectively in two groups of non transfused patients, one group treated with leukocyte-free filtered red cells, the other with buffy-coat-free packed red cell units. Different types of antileukocyte antibodies were looked for in both groups and the results were compared. In study II the behavior of pre-existing lymphocytotoxic antibodies found in the serum of children previously transfused with standard or buffy-coat-free packed red cell units was followed after the patients had been passed to a program of transfusion with leukocyte-free filtered red cells. Study I showed that none of the patients transfused with leukocyte-free filtered red cell units have produced antileukocyte antibodies, while these could be found in 2/3 of the patients transfused with buffy-coat-free packed red cell units. Study II showed that the repeated transfusion of leukocyte-free filtered red cells to patients who possessed in their serum preformed lymphocytotoxic antibodies did not cause any increase in the potency or spectrum of these antibodies, but was in fact accompanied in some cases by their decrease or disappearance. It is concluded that filtration through cotton is an easy and inexpensive means of preparing leukocyte-free red blood cells for transfusion capable of preventing (or reducing) the production of antileukocyte antibodies in multitransfused patients.

Antilymphocyte Serum↗

An immunoenzymatic assay for the detection and quantitation of platelet antibodies: the platelet beta-galactosidase test (PGT).

An immunoenzyme assay for detection of platelet antibodies and suitable for routine use is described. Purified rabbit IgG anti-human IgG antibodies are conjugated to beta-galactosidase with meta-maleimidobenzoyl-hydroxysuccinimide ester as a bifunctional reagent and o-nitrophenyl-beta-galactopyranoside as a substrate to evaluate the enzymatic activity of the labeled antiglobulin. The sera of 26 patients suffering from various diseases (acute leukemia, aplastic anemia and systemic lupus erythematosus) and 40 control subjects were assayed with the enzyme-labeled reagent and, for comparison, with an indirect immunofluorescence technique. Half of these patients had never been transfused. Platelet antibodies were detected by both assays in all the transfused patients except one, and in 3 out of 13 non-transfused patients. The sera of all the control subjects were negative. Quantitation of platelet antibodies was obtained by a sensitive antiglobulin absorption technique. A method for standardization of the reagents allowing comparison of results obtained in the same patient at different times and suitable for long-term follow-up studies is also described.

Absorption↗

The first stable variant of erythrocyte glucose-phosphate isomerase associated with severe hemolytic anemia.

A new variant of glucosephosphate isomerase (GPI) associated with hemolytic anemia, mental retardation, and muscular hypotonia is described. The defective enzyme showed increased affinity for fructose-6-phosphate (F-6-P), decreased affinity for glucose-6-phosphate (G-6-P) altered electrophoretic and isoelectrofocusing patterns, and shift to the left of the precipitin curve. The enzyme was stable under all the conditions tested (heat, urea, guanidine-HCl, and storage). Optimum pH, Ki for 6-phosphogluconic acid (6-PGA) and for erythrose-4-phosphate (E-4-P), molecular weight, GPI-related antigen concentration, immunodiffusion pattern, and immunoinactivation were in the normal range. This is the first example of the association of a stable mutant GPI with severe hemolytic anemia. Enzyme instability has been present in all previously reported cases.

Anemia, Hemolytic, Congenital↗

Acidified glycerol lysis test: a screening test for spherocytosis.

Acidified glycerol lysis test (AGLT) is a screening procedure which has been developed for spherocytosis. AGLT was found positive in 100% of 48 patients suffering from hereditary spherocytosis, 100% of nine couples of affected parents, and 86% of 14 couples of clinically healthy parents. The test was positive in acquired spherocytosis and negative in normal controls. AGLT appears to have a predictive value higher than the entire battery of conventional tests. It is simple, rapid, inexpensive, gives clear-cut results and requires minute amounts of blood. It can be performed on blood stored for up to 24 h.

Adult↗