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Start Over Author "Perugini L" Journal haematologica
3 results on '"Perugini L"'

1. Recovery of cord blood hematopoietic progenitors after successive freezing and thawing procedures

Author

Timeus, F, Crescenzio, N, Saracco, P, Doria, A, Fazio, L, Albiani, R, CORDERO DI MONTEZEMOLO, Luca, Perugini, L, and Incarbone, E.

Subjects
Cryopreservation, Cell Survival, Feasibility Studies, Humans, Cord Blood Stem Cell Transplantation, Fetal Blood, Hematopoietic Stem Cells, Blood Cell Count
Abstract

Cord blood (CB) is a valuable source of stem cells. Most CB units are still cryopreserved in single bags in the world's CB banks. Thawing a single CB unit, dividing it into two parts, expanding the smaller one and refreezing the other would optimize ex vivo expansion of CB progenitors prior to transplantation: expanded and unexpanded cells could be infused together to accelerate early engraftment.The feasibility of refreezing CB samples was investigated by evaluating the effect of 3 successive cryopreservation procedures in 9 CB units. The number and viability of WBC, BFU-E, CFU-GM, CFU-MIX, LTC-IC, and the absolute CD34+ cell count were assessed at time 0 and after each thawing. The percentage of CD34 cells expressing CD38, L-selectin, VLA-4, VLA-5, H-CAM, LFA-1 and CXCR4 was also evaluated.After three freezing and thawing procedures, WBC counts decreased, while lymphocytes were unchanged. Viability was 90% of basal values after the first thawing and did not change. BFU-E decreased significantly only after the third thawing. CFU-GM and CFU-MIX did not change significantly, nor did LTC-IC, CD34+ cell counts and CAM and CXCR4 expression on CD34+/ CD38-- cells.These data show that two successive freeze-thaw procedures do not significantly affect the clonogenic potential and CAM expression of cord blood progenitors. This information could be exploited to devise new options in ex vivo expansion procedures and quality controls prior to transplantation.

Published
2003

2. Carrier detection for prenatal diagnosis of hemophilia A in Italian families.

Author

Cappello N, Restagno G, Garnerone S, Gennaro C, Perugini L, Rendine S, Piazza A, and Carbonara A

Subjects
Alleles, DNA Mutational Analysis, Factor VIII analysis, Female, Fetal Diseases diagnosis, Fetal Diseases genetics, Genetic Markers, Haplotypes, Hemophilia A diagnosis, Hemophilia A epidemiology, Hemophilia A genetics, Humans, Incidence, Italy epidemiology, Lod Score, Male, Pedigree, Polymorphism, Restriction Fragment Length, Risk, Software, Genetic Carrier Screening, Hemophilia A prevention & control, Prenatal Diagnosis
Abstract

Background: The results obtained from a comparative analysis between phenotypic bioassays as the ratio of factor VIII: C clotting activity to factor VIII: C-related antigen, and DNA haplotypes from RFLP's TaqI/St14 and BclI/F8A in 12 hemophilia A (HeA) families are described., Methods: DNA from HeA patients and related at-risk women has been analyzed by Southern blotting with two probes: the intragenic F8A and the extragenic St14. Factor VIII: C coagulant activity was measured by a one-stage method, and the Factor VIII-related antigen (FVIII: RAg) was assayed with bidimensional electrophoresis. Linkage analysis was performed with the LINKAGE computer programs; in particular, the risks of carrying HeA were calculated using the MLINK program., Results: The observed heterozygosity for the flanking marker DXS 52 (TaqI/St14 RFLP) in combination with intragenic BclI/F8A polymorphism was 0.94. A statistically significant difference in frequency was detected at the DXS 52 locus (allele 4) in comparison with other Caucasian populations. Linkage analysis made it possible to combine the plasma bioassay values with the DNA marker haplotypes to determine the probability of carriership; 22 females at risk were investigated: 4 of them were identified as carriers and 18 were excluded. The risk of carrying hemophilia A for some women at risk in six families is reported., Conclusions: This study compares a classic method and DNA analysis in genetic counselling for hemophilia A. In some cases the two methods may give different results when identifying carriers in at-risk families. From these data it is possible to conclude that DNA analysis combined with the phenotypic bioassays for carrier detection gives more information that the two analyses taken separately.

Published
1992

3. Pyrimidine 5'-nucleotidase and oxidative damage in red blood cells transfused to beta-thalassemic children.

Author

David O, Sacchetti L, Vota MG, Comino L, Perugini L, and Pescarmona GP

Subjects
5'-Nucleotidase deficiency, Blood Transfusion, Child, Child, Preschool, Creatine blood, Erythrocyte Aging, Erythrocyte Transfusion, Glucosephosphate Dehydrogenase blood, Glutathione blood, Humans, Lipid Peroxidation, Oxidation-Reduction, Pentose Phosphate Pathway, Pyruvate Kinase blood, Thalassemia therapy, 5'-Nucleotidase blood, Erythrocytes enzymology, Thalassemia blood
Abstract

Pyrimidine 5' nucleotidase (P5'N) acquired deficiency has been found in several hematologic disorders including beta-thalassemia. Our previous studies suggested that the aldehydes produced during membrane lipid peroxidation could play a role in P5'N inactivation in thalassemia. To evaluate the effects of the thalassemic "environment" on transfused red blood cells, we tested P5'N, pyruvate kinase (PK), glucose 6-phosphate dehydrogenase (G-6PD) activity, creatine content, reduced glutathione (GSH) levels and the hexose monophosphate shunt (HMS) in the red cells of homozygous transfusion-dependent thalassemic children, immediately following and again one month after transfusion. In red cells aged in thalassemic plasma, P5'N activity, creatine level, GSH stability and stimulated HMS flux were significantly decreased. These results fit in with the presence in thalassemic plasma of molecules interfering with antioxidant red cell defenses. Normal red cells incubated in thalassemic plasma display a significant stimulation of the basal HMS (p less than 0.01). Transfused red cell metabolic alterations could be explained by the plasma pro-oxidant activity and may contribute to reducing red cell survival in the host plasma.

Published
1990

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