摘要:

AbstractPrenatal diagnosis was carried out on a woman who had previously given birth to a son with a spontaneous mutation of C → T transition at nt 31133 of the factor IX (F.IX) gene. The diagnosis was performed on chorionic villi sampling by the method of amplification‐created restriction site (ACRS). It revealed a female fetus with a normal F.IX gene, as confirmed by DNA sequencing after delivery. Meanwhile, a survey using the ACRS method to evaluate the inheritance of 63 individuals from 8 hemophilia B families was done. A different single‐point mutation in each family was proved by DNA sequencing. One individual had a mutation with a naturally‐created restriction site. In each of the remaining patients, we were able to show an enzyme‐cutting site in their DNA amplification product for ACRS with the designed mutagenesis primers. All patients and carriers could be diagnosed accurately by comparing ACRS results with clinical and laboratory findings. There were new novel mutations among the patients. © 1996 Wiley

ISSN:0361-8609    

DOI:10.1002/(SICI)1096-8652(199608)52:4<243::AID-AJH1>3.0.CO;2-S

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1996

数据来源: WILEY

摘要:

AbstractType 2B von Willebrand disease is characterized by an abnormal von Willebrand factor molecule with increased affinity for the platelet glycoprotein (GP) lb‐IX receptor. A diagnostic feature of type 2B von Willebrand disease is a characteristic loss of von Willebrand factor high molecular weight multimers. In vitro, the soluble interaction of normal von Willebrand factor with platelets can be initiated with exogenous modulators, the most common being the antibiotic ristocetin. The variant molecules resulting in type 2B von Willebrand disease can sustain binding to platelets at subnormal levels of ristocetin. We characterized the von Willebrand factor gene of an individual with type 2B von Willebrand disease and identified a nucleotide transition resulting in an Arg543→Trp amino‐acid substitution within the GP lb‐IX binding domain of von Willebrand factor. In this study we demonstrate that a recombinant plasmid capable of expressing the isolated GP lb‐IX binding domain of von Willebrand factor, and containing the Arg543→Trp amino‐acid substitution, secretes a dimeric molecule that supports platelet agglutination using subnormal levels of ristocetin. These results demonstrate that the mutation at position 543 increases the affinity between the variant molecule and platelet GP lb‐IX as an intrinsic feature of the isolated von Willebrand factor domain. Thus, structural perturbations within the GP lb‐IX binding domain that are independent of the von Willebrand factor multimeric structure can sufficiently increase the affinity of von Willebrand factor to sustain platelet aggregation, using subnormal levels of ristocetin. © 19

ISSN:0361-8609    

DOI:10.1002/(SICI)1096-8652(199608)52:4<248::AID-AJH2>3.0.CO;2-S

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1996

数据来源: WILEY

摘要:

AbstractThe sciatic nerve of C57Bl mice was examined with a transmission electron microscope to study the ultrastructural alterations in Schwann cells following treatment with escalating doses of vincristine. Results indicated that the drug exerts a dose‐related effect. Total doses up to 8 µg/mouse did not cause any visible damage to Schwann cells. Higher doses induced not only damage to individual cells, but also affected a greater percentage of them. The myelin sheath was the most affected organelle. Schwann cells of myelinated fibers showed greater damage than those of unmyelinated fibers. © 1996 Wiley‐Liss

ISSN:0361-8609    

DOI:10.1002/(SICI)1096-8652(199608)52:4<254::AID-AJH3>3.0.CO;2-R

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1996

数据来源: WILEY

摘要:

AbstractUsing a reverse transcription‐polymerase chain reaction (RT‐PCR) technique we determined the α2/α1, α/β, and γ/β mRNA ratios in reticulocytes of 11 patients with seven different unstable β chain variants, of 4 patients with two unstable α chain variants, in hemoglobin (Hb) D, Hb Porto Alegre, and Hb E heterozygotes, and in 8 patients with Hb X‐β°‐thalassemia (thal) (three D‐β°‐thal, one Porto Alegre‐β°‐thal, one Lulu Island‐β°‐thal, and three E‐β°‐thal). In addition, we determined the βx/βAmRNA ratios (X = unstable) in some Hb D heterozygotes and in 6 subjects with an unstable β chain variant. Normal α/β and βx/βAmRNA ratios were found in all heterozygotes tested, indicating that the respective mutations did not alter the stability of the mRNAs. The α/β mRNA ratio in four Hb E heterozygotes averaged 4.21 (normal, 4.47, and that in 2 patients with Hb E‐β°‐thal and four α‐globin genes (αα/αα) averaged a high 22.4. The γ mRNA level in the Hb E heterozygotes was<1% but varied greatly in patients with Hb E‐β°‐thal; the α/(γ + β) mRNA ratios in the 2 patients were 15.5 and 16.7, respectively. The large differences in α/β and α/(γ + β) mRNA ratios in reticulocytes of subjects with AE and with E‐β°‐thal may be due to differences in the levels of normally‐spliced βEand abnormally‐spliced βEmRNAs. Only the latter is unstable and is preferentially produced in bone marrow and re

ISSN:0361-8609    

DOI:10.1002/(SICI)1096-8652(199608)52:4<258::AID-AJH4>3.0.CO;2-R

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1996

数据来源: WILEY

摘要:

AbstractTo characterize the persistent abnormalities of hematopoiesis in aplastic anemia (AA) after immunosuppression with antilymphocyte globulin (ALG), we analyzed the quantity, phenotype, and growth properties of hematopoietic progenitor cells in 13 patients who received ALG treatment. Flow cytometry (FACS) revealed a deficiency of CD34+cells in bone marrow (BM) of all patients. This deficiency was most severe (40‐fold) in 4 patients in AA relapse. In 9 patients in remission, CD34+cells were reduced 2–10‐fold and showed no correlation with the ALG‐induced improvement of peripheral blood cell counts. The proportion of CD34+cells carryingc‐kitreceptors was abnormally low (2–10‐fold below normal) in 5 of 13 AA patients. These patients also displayed low levels ofc‐kitmRNA by reverse transcription‐polymerase chain reaction (RT‐PCR). Furthermore, the CD34+cell population was almost completely depleted of CD34+CD38‐early hematopoietic progenitors in all AA patients. The proportion of CD34+cells expressing lineage differentiation antigens CD33, CD71, and CD45RA in AA was increased, as compared to control BM. Formation of hematopoietic colonies by FACS‐purified CD34+cells was nearly absent in 4 relapsed patients, normal in 4 of 9, and decreased (up to 10‐fold) in 5 of 9 patients in remission. The degree of impairment of colony‐forming ability by AA progenitors correlated well with the reduction of CD34+c‐kit+cells. The best proliferative response of CD34+cells was observed in the presence of stem cell factor and, in some cases, flt3 ligand. Our results indicate that the disease process in AA depletes immature BM progenitors, thus providing a plausible explanation for persistent defects in colony‐forming ability and long‐term regenerative capacity of AA marrow after immunosuppression. Analysis of the immunophenotypes and the proliferative properties of purified progenitors may be useful for estimating degree of hematopoietic recovery in ALG‐treat

ISSN:0361-8609    

DOI:10.1002/(SICI)1096-8652(199608)52:4<264::AID-AJH5>3.0.CO;2-Q

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1996

数据来源: WILEY

摘要:

AbstractAn intensive brief chemotherapy and radiotherapy regimen including high doses of cyclophosphamide (5 g/m2), etoposide (1 g/m2), epirubicin (180 mg/m2), and ifosfamide (5 g/m2) administered in a period of 30 days followed by involved field radiotherapy to sites of initial bulky disease was administered to 46 untreated patients with high‐intermedium and high‐risk malignant lymphoma. G‐ or GM‐CSF were used as hematological support instead of bone marrow transplantation. All patients had more than 3 adverse prognostic factors at diagnosis.Forty‐one patients (89%) achieve complete response (33 after chemotherapy and 8 partial responses were converted to complete response after adjuvant radiotherapy). Actuarial failure‐free survival at 3 years is 83% and 37 of all patients started on therapy remain alive and in first remission at a median of 24.3 months from completion of treatment. Nearly all patients developed granulocytopenia grade IV; only 13 episodes of bacterial infection were documented. Because hematological recovery was very short (mean 13.6 days) no death related treatment and opportunistic infections were observed. Other non‐hematological toxicities were scarce and well tolerated. No decrease>10% was observed in the left ventricular ejection fraction. None have developed clinically evident congestion heart failure or other late side effects. These results showed that G‐ or GM‐CSF can act as hematological support instead of bone marrow transplantation during intensive and brief chemotherapy. These regimens produce higher complete remission rate, and adjuvant radiotherapy will improve the outcome in patients with bulky disease. © 19

ISSN:0361-8609    

DOI:10.1002/(SICI)1096-8652(199608)52:4<275::AID-AJH6>3.0.CO;2-P

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1996

数据来源: WILEY

摘要:

AbstractChimerism can be monitored after HLA‐matched allogeneic bone marrow transplantation (BMT) or allogeneic peripheral blood stem cell transplantation (PBSCT) by detecting polymorphisms in short tandem repeats (STR). The purpose of our study was to document early complete chimerism in BMT and PBSCT recipients using STR, and to determine whether the initial WBC recovery correlated with the days required to attain complete chimerism. A total of 5 patients (2 PBSCT and 3 BMT) were followed by STR after transplantation. Peripheral blood obtained prior to transplantation was used to determine the 2 most informative STR probes for each donor/recipient pair. STR were amplified by polymerase chain reaction (PCR) with 8 commercial probes, and PCR products were visualized with silver staining. Peripheral blood was evaluated daily post‐transplantation for WBC counts and to identify the presence of mixed or full chimerism by STR. The sensitivity of the STR technique varied from 0.05 to 1%, depending on the probe. Full chimerism was documented between day 9 and 14 in PBSCT recipients and on day 14 and 16 in BMT recipients. The initial rise in WBC occurred within 3 days of the onset of full chimerism, indicating that full chimerism is a more sensitive indicator of early engraftment. Periodic recipient monitoring using STR after complete chimerism identifies those patients who revert to mixed chimeras. The STR method may be useful in future studies to determine the significance of early engraftment and the clinical implications of sustained complete chimerism or mixed chimerism. © 1996 Wiley‐Lis

ISSN:0361-8609    

DOI:10.1002/(SICI)1096-8652(199608)52:4<281::AID-AJH7>3.0.CO;2-O

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1996

数据来源: WILEY

摘要:

AbstractContrary to a recent report [Rinder et al.:Blood82:505, 1993], aspirin does inhibit the release of α‐granule contents as well as inhibiting the release of dense granule contents by human platelets during ADP‐induced aggregation in citrated platelet‐rich plasma (PRP). Measurements were: percent release of14C‐serotonin from prelabeled platelets, radio‐immunoassay of β‐thromboglobulin (βTG), and expression on the platelet surface of the α‐granule constituent, P‐selectin, by flow cytometry. During the second phase of ADP‐induced aggregation, 69.0 ± 8.3% of βTG and 54.1 ± 4.6% of14C‐serotonin were released (means ± SEM, n = 13); aspirin treatment reduced these values to 6.0 ± 1.2 and 1.0 ± 0.3%, respectively. In contrast, incubation of platelets with ADP without stirring caused only 6.7 ± 1.7% release of βTG and 2.1 ± 0.4% release of14C‐serotonin; these low values were not appreciably affected by aspirin. During ADP‐induced primary aggregation in PRP anticoagulated with FPRCH2Cl (PPACK), only 4.7 ± 0.9% release of βTG and no detectable release of14C‐serotonin occurred; aspirin had no effect. In both stirred and unstirred PRP, the thrombin receptor activating peptide, SFLLRN (50 μM), caused at least 75% release of the contents of both granules, which was partially inhibited by aspirin. Upon incubation of platelets with ADP (2–10 μM), the mean fluorescence intensity due to P‐selectin was<14% of that induced by SFLLRN. In this unstirred system used for flow cytometry, aspirin treatment caused no significant inhibition of P‐selectin expression. Thus, under conditions in which ADP does not cause secondary aggregation (physiological Ca2???concentration or unstirred citrated PRP) release of the contents of both types of granules is less than 7% and aspirin is not inhibitory; the P‐selectin expression associated with this low percent release is also unaffected by aspirin. However, aspirindoesstrongly inhibit the extensive release of both α‐granule and dense granule contents during ADP‐induced second

ISSN:0361-8609    

DOI:10.1002/(SICI)1096-8652(199608)52:4<288::AID-AJH8>3.0.CO;2-O

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1996

数据来源: WILEY

摘要:

AbstractRare inherited cancer syndromes have proven invaluable for the identification of genes involved in the more frequent corresponding noninherited cases. We report on a family with an adult onset, incompletely penetrant, autosomal dominant syndrome of myelodysplasia and acute myelogenous leukemia, affecting at least eight, and probably ten, individuals from three generations. The patients have developed leukemias differing in morphologic subtype, tumor cytogenetics, and abruptness of presentation. Some have presented with acute onset and others with protracted myelodysplasia. This family does not have an unusual incidence of other malignancies; however, one person at 50% risk of inheriting this gene developed atypical mycobacterium infection in the absence of leukemia, but also without appreciable risk factors for acquired deficiencies in cellular immunity. Features common to affected family members, including the individual with mycobacterium infection, are the early presence in the bone marrow of red cell and platelet maturation defects. A search for mutations in diseased marrows fails to detect abnormalities ofp53or N‐ras. Two of the affected family members, third degree relatives, have co‐inherited a constitutional chromosomal banding variation of 9p21–22, potentially suggesting linkage to this locus. The variable penetrance and expressivity of this syndrome support a multistep model of leukemia evolution, in which the gene defined by this family's syndrome is the signal step. © 1996 Wiley‐L

ISSN:0361-8609    

DOI:10.1002/(SICI)1096-8652(199608)52:4<295::AID-AJH9>3.0.CO;2-N

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1996

数据来源: WILEY

摘要:

AbstractA 13‐year‐old African‐American female with erythrocytosis and three different β globins on electrophoresis βA, βS, and βOsler, raised the possibility that one chromosome 11 might contain a duplicated β globin gene, since there are normally only 2 β globin genes. DNA sequence analysis showed GTG at codon 6 in exon 1, corresponding to Hb S and AAT at codon 145 in exon 3, indicating a substitution of Asn for Tyr. Thus, Hb Osler undergoes spontaneous post‐translational deamidation, β 145 Asn→β 145 Asp. Unmodified Hb Osler (Asn) co‐migrates with Hb A on electrophoresis and co‐elutes with Hb A on HPLC; therefore it has not been identified previously. All previous studies have incorrectly identified the mutation as being β 145 (HC 2) Tyr→Asp

ISSN:0361-8609    

DOI:10.1002/(SICI)1096-8652(199608)52:4<305::AID-AJH10>3.0.CO;2-C

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1996

数据来源: WILEY