Güneş T. Yüregir

βs haplotypes in various world populations

SummaryWe have determined the βs haplotypes in 709 patients with sickle cell anemia, 30 with SC disease, 91 with S-β-thalassemia, and in 322 Hb S heterozygotes from different countries. The methodology concerned the detection of mutations in the promoter sequences of the Gγ- and Aγ-globin genes through dot blot analysis of amplified DNA with 32P-labeled probes, and an analysis of isolated Hb F by reversed phase high performance liquid chromatography to detect the presence of the AγT chain [Aγ75 (E19) Ile→Thr] that is characteristic for haplotype 17 (Cameroon). The results support previously published data obtained with conventional methodology that indicates that the βs gene arose separately in different locations. The present methodology has the advantage of being relatively inexpensive and fast, allowing the collection of a vast body of data in a short period of time. It also offers the opportunity of identifying unusual βs haplotypes that may be associated with a milder expression of the disease. The numerous blood samples obtained from many SS patients living in different countries made it possible to compare their hematological data. Such information is included (as average values) for 395 SS patients with haplotype 19/19, for 2 with haplotype 17/17, for 50 with haplotype 20/20, for 2 with haplotype 3/3, and for 37 with haplotype 31/31. Some information on haplotype characteristics of normal βA chromosomes is also presented.

Molecular characterization of β-thalassemia in Azerbaijan

We have analyzed the β-thalassemia mutations in 99 chromosomes of 49 adults with β-thalassemia major and of one with Hb S-β-thalassemia, who are regular patients at a large hematology clinic in Bakü, Azerbaijan. A total of 20 different mutants were identified; three [frameshift at codon 8 (-AA); IVS-II-I (G→A); IVS-I-110 (G→A)] were present in about two-thirds of all chromosomes. Most alleles are the same as found in Mediterranean populations; a few have an Asian origin or come from Kurdistan, Lebanon, Saudi Arabia, or a black population. One mutant [frameshift at codons 82/ 83 (-G)] might be specific for the Azerbaijanian population. Nearly all patients were transfused, which made quantitation of Hb F impossible; highGγ values were present in the Hb F of those patients whose β-thalassemia chromosome carried the C → T mutation at position — 158 in the promoter of the Gγ-globin gene.

Studies on Red Cell Glucose-6-Phosphate Dehydrogenase: Evaluation of Reference Values

The analytical, intra-individual, inter-individual variation and reference values were determined for red cell glucose-6-phosphate dehydrogenase (G6PD). Different procedures for the conditions for storage of red blood cells and the preparation of haemolysates were investigated. A total of 2170 samples of blood were taken from apparently healthy persons—1212 males and 958 females—from randomly selected villages and city centres in the southern part of Turkey. Analytical variation, intra-individual variation and inter-individual variation were 8·67%, 32·8% and 31·8%, respectively. The mean (SD) for G6PD was 8·6 (3·3) IU/gHb. The index of individuality, 1·03, showed that the reference intervals could be used for diagnostic purposes. Whole blood or a red cell pellet could be stored in physiological saline for one week at 4°C or −20° with little loss of activity. Two of three different procedures for the preparation of haemolysate gave data that showed no statistical difference and were equally satisfactory.

GENETIC HETEROGENEITY OF β-THALASSEMIA AT ÇUKUROVA IN SOUTHERN TURKEY

β-Thalassemia is the most common genetic abnormality causing health problems worldwide.Ç ukurova, in the southern part of Turkey, being on the Mediterranean, is in the thalassemic belt. Since there is no cure for the disease at present, the frequency of the mutation types of β-thalassemia must first be identified to aid in clinical follow-up and prenatal diagnosis. Carriers identified during a screening survey and patients referred to our laboratory were studied for this purpose. After routine hematological analysis molecular screening was performed by the amplification refractory mutation system and DNA sequencing. The frequency of the common mutations were: IVS-I-110 (G→A) 57.3%, IVS-I-1 (G→A) 8.3%, codon 39 (C→T) 6.4%, IVS-I-6 (T→C) 5.7%, frameshift codon 8 (–AA) 5.7%, –30 (T→A) 4.7%, IVS-II-1 (G→A) 3.4%, IVS-II-745 (G→C) 2.8%, and frameshift codon 5 (–CT) 1.1%. Some rare mutations (1%) such as frameshift codon 44 (–C) 0.7%, frameshift codons 74/75 (–C) 0.7%, IVS-I-5 (G→C) 0.7%, frameshift codons 8/9 (+G) 0.4%, frameshift codons 36/37 (–T) 0.4%, frameshift codons 22/23/24 (–AAGTTGG) 0.4%, IVS-I-130 (G→C) 0.4%, IVS-I-5 (G→T) 0.2%, –28 (A→C) 0.2%, codon 15 (TGG→TGA) 0.2%, and frameshift codons 82/83 (–G) 0.2%, were detected by sequence analysis. The codon 15 (TGG→TGA) and frameshift codons 82/83 (–G) mutations were seen in Turkey for the first time.

Evaluation of reference values for erythrocyte glutathione

The analytical, intra-individual and inter-individual variations as well as the best storage conditions were determined for erythrocyte glutathione, and the reference values were established. A total of 396 apparently healthy people, 206 male, and 190 female, were randomly selected from villages and cities of the southern part of Turkey. The distribution was Gaussian and no significant difference was observed between the male and the female subjects. The mean (standard deviation) of the population investigated for glutathione was 6.9 (1.0) mumol/gHb. The analytical, intra-individual and inter-individual variations were assessed in 20 apparently healthy subjects and were found to be 4.63%, 13.67% and 11.16%, respectively. Whole blood stored at -70 degrees C for up to 10 days was shown to be the best storage condition for erythrocyte glutathione determination. The results of the index of individuality showed that glutathione reference values could be used for diagnostic purposes.

Intracellular glutathione content in leukemias

The intracellular glutathione (GSH) content was measured in 73 patients with leukemia and compared with controls. GSH content was between 1.16 and 5.55 mumol/g protein (mean 2.96 +/- 0.86) in the study group and between 0.5 and 1.48 mumol/g protein (mean 1.31 +/- 0.27) in the control group, statistically significant difference (p = 0.0000). There was no significant difference between acute and chronic leukemias, lymphoid and myeloid leukemias and, more importantly, newly diagnosed and relapsed patients. GSH content did not change significantly with clinical and hematologic parameters such as age, sex, and initial hematologic findings. In addition, variable changes were detected over 24 h in 9 patients. It can be concluded that GSH content in leukemic cells was higher than in controls and showed a wide range. The absence of a relationship between GSH content and clinical and laboratory parameters suggested that GSH is not the sole determinant of response to cytotoxic drugs. GSH variation over a 24-hour period may be important in the timing and success of chemotherapy for leukemias.

Glucose 6-Phosphate Dehydrogenase Deficiency both in Red Blood Cells and Lenses of the Normal and Cataractous Native Population of Çukurova, the Southern Part of Turkey

Glucose 6-phosphate dehydrogenase (G6PD) activity was measured in both red blood cells and lenses of persons with cataracts living in Cukurova, the southern part of Turkey, where G6PD deficiency is well documented. The incidence of red blood cell G6PD deficiency among the patients with cataract (33.3%) was significantly higher than that in individuals with clear lenses (8.2%). The incidence of lens G6PD deficiency in a total of 52 patients with cataract was 52%. Of the lens G6PD-deficient cases, 46.2 and 28.6% of female and male patients, respectively, also showed red blood cell G6PD deficiency.

Prenatal diagnosis of Hb H disease caused by a homozygosity for the alpha2 poly A (AATAAA-->AATAAG) mutation.

Institute of Molecular Medicine and Genetics, Room CB-2803, 1120 15th Street, Medical College of Georgia, Augusta, GA 30912 USA Department of Pediatric Hematology, CË ukurova University Medical Faculty, 01330 Adana, Turkey Department of Gynecology and Obstetrics, CË ukurova University Medical Faculty, 01330 Adana, Turkey Department of Biochemistry, CË ukurova University Medical Faculty, 01330 Adana, Turkey

Three new G6PD variants, G6PD Adana, G6PD Samandağ, and G6PD Balcali in Cukurova, Turkey.

SummaryGlucose-6-phosphate dehydrogenase (G6PD) enzyme from cases known to be completely or mildly deficient were analyzed. The enzymes were purified from blood samples by utilizing DEAE-52 cellulose pH 7.0 column chromatography and ammonium sulphate precipitation. Biochemical and electrophoretic properties of G6PD were studied in these partially purified enzymes. In this study wer report three new variants from Çukurova, named Adana, Samandağ, and Balcali. Variant I (G6PD Adana) had a high Km for G6P (210 μM) and NADP (13μM). Utilization of 2d-G6P was 38%. It had a slow electrophoretic mobility, a biphasic pH optimum curve, and abnormal heat stability. Variant II (G6PD Samandağ) had a low Km for G6P (25μM) and a high Km for NADP (18μM). The rate of utilization of 2d-G6P was normal. G6PD Samandağ deviated from the normal enzyme by its biphasic pH optimum curve and its slow electrophoretic mobility. Variant III (G6PD-Balcali) had a normal Km G6P, NADP and rate of utilization of 2d-G6P. However, it showed a biphasic pH optimum curve and slow electrophoretic mobility.

Serum copper and zinc values compared with serum iron, total iron-binding capacity, and transferrin saturation in sickle cell trait : A preliminary report.

Reliable data for the trace element values of the biological systems in some diseases are still very rare. Sickle cell trait is one of them.For this purpose, serum iron, zinc, and copper values, together with the total iron binding capacity and saturation percent, were determined in cases with sickle cell trait, eliminating all the sources contributing to deviations from the normal values by choosing a control group from the relatives of the cases.In this study, the values of two groups were compared on the basis of the difference in hemoglobin type, which was the only parameter affecting the trace element analysis.