Characterization of Genetic Variants of Blood Proteins

Smithies, O.

doi:10.1159/000384432

Cited by 31 publications

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“…Aliquots of 3 ml of blood were directly analyzed using the model 1312 Automatic Blood Gas Manager (Instrumentation Laboratory, Usselstein). Red cell lysates were examined on starch gel electrophoresis at pH 8.6 [23] and on isoelectrofocusing, as previously described [9]. The Hb A 2 fraction was estimated by ion-exchange column chromatography [1].…”

Section: Methodsmentioning

confidence: 99%

Hb Malmö [β-97(FG-4)His→Gln] leading to polycythemia in a Dutch family

Giordano

Harteveld

Brand

et al. 1996

Annals of Hematology

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We have examined six individuals from a two-generation Dutch family for a suspected hemoglobin (Hb) abnormality. The propositus presented with polycythemia and complained of persistent weakness, headache, and epistaxis. All family members initially showed a normal Hb-electrophoretic pattern, but on isoelectric focusing, three of them displayed a fast-moving band associated with high packed red cell volumes (PCV) and increased red blood cell count. The Hb mutant was analyzed at the DNA level by specific gene fragment amplification (PCR), followed by direct DNA sequencing, and the mutation was confirmed by restriction enzyme analysis. We found a C-->G transversion (CAC-->CAG) at codon 97 of the beta-chain, which corresponded to the His-->Gln amino acid substitution previously described as Hb Malmö. We report here the clinical history of the patient, the effects of phlebotomy treatment, and the effect of subnormal iron conditions on the erythropoietic recovery after phlebotomy. The mechanism responsible for the induction of the higher oxygen affinity is discussed, as are some aspects concerning the occurrence, pathology treatment, and the genetic risk of Hb variants with high O2 affinity.

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Section: Methodsmentioning

confidence: 99%

Hb Malmö [β-97(FG-4)His→Gln] leading to polycythemia in a Dutch family

Giordano

Harteveld

Brand

et al. 1996

Annals of Hematology

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“…The in vitro synthetic ratio between the α-and non α-globin chains was measured according to Weatherall et al (1969) using a modified system, which includes the elimination of white cells and the enrichment of the reticulocyte fraction. Red cell lysates were examined by starch gel electrophoresis at pH 8.6 to test for the presence of hemoglobin variants (Smithies 1965).…”

Section: Methodsmentioning

confidence: 99%

α-Thalassemia in The Netherlands: a heterogeneous spectrum of both deletions and point mutations

Harteveld

Losekoot²,

Heister

et al. 1997

Human Genetics

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In this article we describe the molecular characterization of 104 independent alpha-thalassemia patients identified by hematological analysis and family studies. During the study, another six chromosomes were identified with rearrangements of the alpha-cluster or point mutations in the alpha 2-globin gene, not associated with alpha-thalassemia, in healthy relatives of the patients. The molecular defects were established by Southern blot analysis and, if no deletions could be identified, the alpha-globin genes were investigated by denaturing gradient gel electrophoresis and single strand conformation analysis for the presence of point mutations. Following this strategy, we were able to identify the molecular basis of 131 independent alpha-thalassemia chromosomes. In two individuals, the alpha-thalassemia determinant could not be demonstrated at the molecular level. We identified eight different deletion and five non-deletion alpha-thalassemias, three rearrangements in the alpha-cluster, two alpha-chain variants, and a silent mutation in the alpha 2-globin gene not associated with alpha-thalassemia. The large heterogeneity of alpha-thalassemia mutations seen in the Dutch population might be typical for northern European countries where, besides the more common mutations introduced by migration, a variety of sporadic mutations was also found in the autochthonous population. The screening strategy as described here, capable of identifying a wide spectrum of both deletions and point mutations, identified 98% of the alpha-thalassemia determinants present in 133 chromosomes.

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“…Routine laboratory procedures were used for haematological investigations [Dacie and Lewis, 1966], haemoglobin H inclusion bodies were demonstrated by incubating erythrocytes in a 1-per cent solution of brilliant cresyl blue for tw'o hours at 37° C. Haemoglobin F was estimated by the method of Singer et al [1951]. Haemoglobin electrophoresis was carried out on starch gel, using a tris ETDA-horate buffer system at pH 8.6 [Smithies, 1965] and further characterization of fast haemoglobins in a phosphate buffer pH 7.0 [Hueiins et al,I960]. Quantitation of A,A, and H was performed after electrophoresis on cellulose acetate at pH 8.6 for 60 min by elution of the different fractions and spectrophotometric estimation [Marengo-Rowe, 1965].…”

Section: Methodsmentioning

confidence: 99%

The Genetical Interpretation of Haemoglobin H Disease

Kattamis¹,

Lehmann²

1970

Hum Hered

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The clinical and haematological data of nine patients with haemoglobin H disease, as well as of 16 members of their families, are reported. Pedigree study and haematological data permit the assumption that in an individual four genes could be responsible for α-chain synthesis. Based on this hypothesis, four variants of α-thalassaemia are expected to exist. The first, in which only one gene is affected has no, or only minor, red cell changes and an increase in Hb Bart’s at birth, the second with two genes affected has in addition Hb H inclusion bodies in a few red cells, the third, with three genes affected represents haemoglobin H disease, and the fourth, with all four genes affected, represents homozygous α-thalassaemia, which is incompatible with life and results in a stillborn child with hydrops foetalis.

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Characterization of Genetic Variants of Blood Proteins

Cited by 31 publications

References 0 publications

Hb Malmö [β-97(FG-4)His→Gln] leading to polycythemia in a Dutch family

Hb Malmö [β-97(FG-4)His→Gln] leading to polycythemia in a Dutch family

α-Thalassemia in The Netherlands: a heterogeneous spectrum of both deletions and point mutations

The Genetical Interpretation of Haemoglobin H Disease

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