Martine Aubry

THROMBOTIC EVENTS IN COMPOUND HETEROZYGOTES FOR A HIGH AFFINITY HEMOGLOBIN VARIANT: Hb MILLEDGEVILLE [α44(CE2)Pro→Leu (α2)] AND FACTOR V LEIDEN

During routine studies of a patient who presented with venous thrombosis and erythrocytosis, a high affinity hemoglobin (Hb) variant, Hb Milledgeville [a44(CE2)Pro!Leu (a2)], was found. The patient was a 57-year-old French Caucasian man and originated from southeast France. He had had deep venous thrombosis, complicated by pulmonary embolism, four years ago. Medical treatment included heparin followed by an oral anticoagulant for four months. After this last treatment, the patient took daily low doses of aspirin. He was referred to us for biological reevaluation. The hematological parameters are shown in Table 1. Neither spleen nor liver enlargement were found at physical examination, and echographic investigations were negative. Bone marrow biopsy ruled out the occurrence of myeloproliferative disorders. The serum erythropoietin level was normal: 11.2 IU=L (normal range: 3.7–16 IU=L). Routine Hb analyses were performed by standard procedures. Cation exchange high performance liquid chromatography (HPLC) using the VARIANT System with the b-Thalassemia Short Program (Bio-Rad Laboratories, Hercules, CA, USA) revealed the presence of an abnormal Hb which eluted as a shoulder just before Hb A, accounting for 23% of total Hb. Isoelectrofocusing

Two New α Chain Variants: Hb Die [α93(FG5)Val→Ala (α1)] and Hb Beziers [α99(G6)Lys→Asn (α1)]

The first new a chain variant here reported, Hb Die [a93(FG5)Val!Ala (a1)], was found during investigations of an anemia prior to surgery for correcting spina bifida. The patient was a 7-year-old girl, living near the town of Die in Southeast France; her parents were of French Caucasian origin. The hematological findings were as follows: RBC 5.8 10/L, hemoglobin (Hb) 11.9 g/dL, MCV 65.5 fL and MCH 20.5 pg; the serum iron level was 6.6 mmol/L (normal values 9–30 mmol/L). Iron treatment associated with erythropoietin was prescribed. The family refused all transfusion therapy. Unfortunately, no hematological data were available after this treatment and no family studies were possible. The second new a chain variant, Hb Beziers [a99(G6)Lys!Asn (a1)], was found during the determination of Hb A1c by high performance liquid chromatography (HPLC) in a diabetic patient. The new Hb variant was detected in a 72-year-old woman, who was also of French Caucasian origin and lived in the city of Beziers in the south of France. The hematological data were as follows: RBC 4.77 10/L, Hb 13.6 g/dL, MCV 86 fL and MCH 28.5 pg; physical examination was normal, without hepatomegaly or splenomegaly.

Hb AUBAGNE [β64(E8)Gly→Ala]: A NEW UNSTABLE β CHAIN VARIANT FOUND IN A FRENCH FAMILY

Only one hemoglobin (Hb) variant has been reported at the internal position 64(E8)Gly of the b-globin chain: Hb J-Calabria, also known as Hb J-Bari and Hb J-Cosenza (!Asp). We here report a new Hb variant at the same position, namely, Hb Aubagne (!Ala), that was found in a young woman suffering from an anemia. The mutation is close to the distal histidine b63(E7) and leads to moderate instability. Routine Hb analyses were performed by isoelectrofocusing (IEF) and high performance liquid chromatography (HPLC) of Hbs and globin chains. Molecular characterization of the mutation was achieved by polymerase chain reaction (PCR), followed by direct DNA sequencing of the PCR products. PCR amplification of exons 1 and 2 of the b-globin gene was carried out using the following primers: 50-TGTACTGATGGTATGGGGC-30 (primer 1) located 225 nucleotides upstream to the Cap site, and 50-AAACGATCCTGAGACTTCC-30 (primer 2) located in the second intron. PCR amplification of exon 3 was performed using: 50-TATCATGCCTCTTTGCACTC-30 (upstream primer 3)

A new alpha chain hemoglobin variant: Hb Al-Hammadi Riyadh [alpha75(EF4)Asp-->Val (alpha2)].

A new hemoglobin (Hb) variant in the heterozygous state, Hb Al-Hammadi Riyadh [codon 75 (GAC→GTC); α75(EF4)Asp→Val (α2)] corresponding to an A→T transversion on the second exon of the α2-globin gene, is described. The variant was characterized by DNA sequencing and mass spectrometry (MS). The variant was found during a routine Hb analysis for anemia in a 16-month-old boy who lived in Riyadh, Kingdom of Saudi Arabia.

Two new hemoglobin variants: Hb Brem-sur-Mer [beta9(A6)Ser-->Tyr] and Hb Passy [alpha81(F2)Ser-->Pro (alpha2)].

Two new hemoglobin (Hb) variants: Hb Brem-sur-Mer [codon 9 (TC T→TA T); β9(A6)Ser→Tyr] on the first exon of the β-globin gene and Hb Passy [codon 81 (T CC→C CC); α81(F2)Ser→Pro (α2)] on the second exon of the α2-globin gene, are described. The two variants were characterized by DNA sequencing and mass spectrometry (MS). Hematological abnormalities: microcytosis and hypochromia were found only in the carrier of Hb Passy. In the absence of an association with an α-thalassemic deletion or mutation, the mutation 81(F2)Pro could induce a possible α-thalassemia (thal).

A Mutation of the β-Globin Gene Initiation Codon, ATG→AAG, Found in a French Caucasian Man

A new mutation of the β-globin gene initiation codon, ATG→AAG (Met→Tyr), is reported in a man originating from the southeast of France. Typical hematological findings of a β-thalassemia (thal) trait were found. We emphasize the importance of characterizing uncommon β-thal mutations for genetic counseling.

Two New α Chain Variants: Hb Part‐Dieu [α65(E14)Ala→Thr (α2)] and Hb Decines‐Charpieu [α69(E18)Ala→Thr (α2)]

We report here the discovery of two new a chain variants that carry the same mutation (Ala ! Thr) located at one turn distance in helix E. The first one, Hb PartDieu [a65(E14)Ala ! Thr (a2)] was found in a systematic screening of hemoglobin (Hb) variants during Hb A1c determination. The patient was a 58-year-old man of French Caucasian origin, living in Lyon in Southeast France. The hematological findings were: RBC 4.3 10/L, Hb 12.6 g/dL, MCV 85 fL and MCH 29.2 pg. The patient suffered from a type 2 diabetes, which recently required insulin treatment. Physical examination revealed hepatomegaly and splenomegaly. Ultrasonic investigations found gall bladder with microlithiasis and portal hypertension. The patient also presented with hypercholesterolemia and hypertriglyceridemia. The second variant, Hb Decines-Charpieu [a69(E18)Ala ! Thr (a2)], was found during laboratory investigations following genetic counseling in a 34-year-old man of French Caucasian origin, who lived in the city of Decines-Charpieu in the south of France. The hematological data were: RBC 4.69 10/L, Hb 14.4 g/dL, MCV

Hb GERLAND [α55(E4)Val → Ala (α2)]: A NEW NEUTRAL α CHAIN VARIANT INVOLVING THE α2 GENE

During routine hemoglobin (Hb) investigations for a possible hemoglobinopathy, performed on a 6-year-old boy suffering from mild microcytic anemia, an abnormal Hb was detected by cation exchange high performance liquid chromatography (HPLC) using the VARIANT System with the b-Thalassemia Short Program (Bio-Rad Laboratories, Hercules, CA, USA). The proband’s parents originate from Southeast France. The abnormal variant, accounting for 31% of the total Hb, eluted as a shoulder 0.3 minutes behind Hb A. The abnormal Hb was not detected by isoelectrofocusing (IEF) or by reversed phase HPLC of globin chains (1–4). Family studies showed the presence of the abnormal Hb, in the same amount, in the proband’s sister and in a lower amount in the father, suggesting a possible interaction with an a-thalassemia (thal) gene. Isopropanol instability test, performed on the proband’s hemolysate, was found to be slightly positive at 30 minutes (5). Physical examination revealed no abnormalities. The hematological parameters of all members of this family are presented in Table 1. Serum iron and serum ferritin levels were normal. Molecular characterization of the mutation was achieved by polymerase chain reaction (PCR), followed by direct DNA sequencing of the PCR products HEMOGLOBIN, 25(4), 417–420 (2001)

Two New β0-Thalassemic Mutations: A Deletion (−CC) at Codon 142 or Overlapping Codons 142-143, and an Insertion (+T) at Codon 45 or Overlapping Codons 44-45/45-46 of the β-Globin Gene

We report here two new β0-thalassemic mutations. In the first case, a deletion of two nucleotides (−CC) at codon 142 was found in a French Caucasian woman. In the second case, an insertion of a single nucleotide (+T) at codon 45 was found in a Turkish girl. In both cases, no dominant thalassemia-like phenotype was observed.

A new G(gamma) chain variant: Hb F-Bron [gamma20(B2)val-->Ala].

A new Gγ hemoglobin (Hb) variant, Hb F-Bron [γ20(B2)Val→Ala] on the first exon of the Gγ-globin gene is described. The variant was characterized by DNA sequencing and mass spectrometry (MS). Hematological abnormalities included hypochromia and microcytosis and were probably caused by an interaction with an α-thalassemia (thal) (3.7 kb) deletion in the heterozygous state.