Kıymet Aksoy

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.

Prenatal Diagnosis of Sickle Cell Anemia and β-Thalassemia in Southern Turkey

Hemoglobinopathies are the most common genetic diseases in Turkey. The incidence of sickle cell trait is 10.0% and β-thalassemia (β-thal) trait is 3.7% in the Çukurova region of southern Turkey. Sickle cell anemia is prevalent in the Çukurova region, but β-thal is seen all over the country. A prenatal diagnosis center was established in 1992 at Adana, Turkey, for the prevention of sickle cell anemia and β-thal. Fifteen hundred and seventy-five fetuses were examined at the Çukurova University Hospital, Adana, Turkey. Three hundred and eighty-six fetuses were diagnosed as homozygous or compound heterozygous for sickle cell anemia and β-thal. A total of 15 different β-thal mutations were characterized in the parents. The incidence of the IVS-I-110 (G > A) mutation accounted for about 50.0% of the parents with β-thal trait. Twenty-four different genotypes were observed in this study. A total of 286 fetuses were diagnosed with homozygous sickle cell disease, 57 fetuses were β-thal homozygotes, 25 fetuses were compound heterozygotes with Hb S [β6(A3)Glu→Val, GAG > GTG], and 18 of the fetuses were double heterozygotes for β-thal mutations.

The effect of immediate decompression on the optic nerve in retrobulbar hematoma

We produced retrobulbar hematoma in both orbits of 10 pigs in order to assess the effects of blood elements and pressure created by the hematoma on the optic nerves. Ten other pigs were used as a control group. Following decompression in the right orbits, ocular movements, fundi, and intraocular pressure were evaluated for 6 weeks. At the end of the 6th week the optic nerves of 20 pigs were dissected bilaterally for measurements of ATP-ase activity and ultrastructural examination. The results of the ultrastructural examination of the optic nerves of the control group were normal. Optic nerves with decompressed retrobulbar hematoma showed minimal degeneration, whereas the nerves subjected to retrobulbar hematoma with no decompression showed significant degenerative changes. For all groups ATP-ase activities were measured and evaluated. Na+, K+ATP-ase activities decreased, while Ca++, Mg++ ATP-ase activities increased with the extent of degeneration. Optic nerve damage can develop after trauma. Decompression procedures are not among the causes of optic nerve degeneration but retrobulbar hematoma can result in optic neuropathy caused by the compression from the hematoma and the direct effect of blood waste products on the optic nerve.

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

Detection of 1311 polymorphism in the glucose-6-phosphate dehydrogenase gene by microarray technique.

Introduction Nucleotide 1311 polymorphism at exon 11 of the glucose-6-phosphate dehydrogenase (G6PD) gene is fairly common in various populations worldwide, especially among Mediterranean populations. In this study, 1311 polymorphism in G6PD-deficient cases was identified by microarray technique. Material and methods Four hundred and fifty clinically healthy subjects were screened and 32 cases were found to have G6PD deficiency (7.11%). Our analysis of genomic DNA samples from 32 G6PD-deficient individuals revealed that the number and percentage of subjects who had a C-to-T alteration at nucleotide 1311 were 21 and 4.7% respectively. Given that the frequency of 1311 polymorphism has been reported in previous studies to be fairly high among G6PD-deficient people with the Mediterranean mutation, our data seem to be inconsistent with what we would expect for this particular region. Results The highly diverse ethnic background of the Adana population which probably results from the high level of immigration into this part of Turkey may be one of the most sensible explanations for this unexpected finding. Nevertheless, it seems that our results need to be confirmed in larger studies. Conclusions The polymorphism studies in the G6PD gene may help us to illuminate the genetic basis of the G6PD deficiency in different regions and in various ethnic groups, and also to discover the influence of a specific polymorphism on the clinical course of the deficiency.

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.

Hemoglobinopathies in the Çukurova Region and Neighboring Provinces.

Abstract To contribute to the creation of a mutation map of the region, we aimed to determine the mutation spectrum of thalassemias and abnormal hemoglobins (Hbs) in the Çukurova region and surrounding provinces. In this study, a total of 8135 samples from Adana, Hatay, Mersin, Konya and Kayseri provinces between 1993 and 2014 were analyzed. Complete blood cell (CBC) counts and Hb typing were carried out using automatic cell counters, cellulose acetate membrane electrophoresis and high performance liquid chromatography (HPLC), respectively. For the molecular analyses, genomic DNA was extracted using both manual and automated DNA extraction devices. Determination of Hb mutations were done by microarray, restriction fragment length polymorphism (RFLP), amplification refractory mutation system (ARMS) and gap-polymerase chain reaction (gap-PCR) methodologies. Samples were analyzed for abnormal Hb and thalassemia mutations. Out of 8135 samples, 1382 were observed to be carrying Hb mutations. It was identified that 826 mutation carriers included abnormal Hbs with a frequency of 59.7%, 416 carriers included β-thalassemia (β-thal) mutations with a frequency of 30.7% and 136 carriers included α-thalassemia (α-thal) mutations with a frequency of 9.9%. In this study, the most frequently observed abnormal Hb in the region was Hb S [β6(A3)Glu→Val (GTG > GAG), HBB: c.20T > A], whereas the most commonly observed mutations were the IVS-I-110 (G > A) (HBB: c.93–21G > A) point mutation in β-thal and the 3.7 kb deletion in α-thal.

Early Ultrastructural Findings and Superoxide Dismutase Levels in Experimental Ischemic Optic Neuropathy: Effect of Hypertension and Hypotension on Ischemic Changes

We induced ischemia, hypertension and hypotension in 15 rabbits in order to evaluate the ischemic changes in the optic nerve and the effect of hypertension and hypotension on ischemia. We cauterized the right internal and external carotid arteries of 15 rabbits and applied dopamine hydrochloride and glycerol trinitrate to 5 each of these rabbits. Two rabbits were used as controls. We enucleated both eyes of all animals at the 24th hour. All of the optic nerves underwent biochemical, histopathological and ultrastructural examination. Histopathological and transmission electron-microscopic changes were found to be more prominent in the hypotensive group. We observed decreased superoxide dismutase levels in all groups, but it was more evident in the third group. In comparison to hypertension, hypotension is found to be a more important factor in the development of early degenerative changes.

Effect of Oxidative Stress on Membrane Proteins in Thalassemia and Iron Deficiency Anemia

ObjectiveTo evaluate the effect of oxidative stress on membrane proteins according to relationship between malondialdehyde (MDA) levels and membrane protein ratios in different two patient groups.MethodsFifty-seven healthy control cases that had no symptoms of inherited blood diseases, 50 diagnosed β-thalassemic patients, and 30 patients with iron deficiency anemia were enrolled. Erythrocyte membrane samples were prepared according to Dodge method and proteins were separated using 8.3% SDS-PAGE. Gels were stained with Coomassie Brillant Blue, then fraction quantities were determined by dansitometric method. MDA levels were measured by thiobarbituric acid method in plasma. One-way ANOVA test was used to compare parameters between groups.ResultsSignificant difference in protein deficiency rates for spectrin and band 4.2 in group I, band 6 in group II and band 3 in group I and II when compared with control group (group III) were found. The significant difference between group I and II wasn’t determined in spite of the fact that MDA levels of patients were different from the control group statistically.ConclusionsAny significant relationship statistically was not found. This consequence may be due to priority removal of oxidatively injured cells from the circulation in thalassemic patients and therefore, significant increase wasn’t determined in the concentration of MDA.

Ultrastructural and Biochemical Effects of Trauma on Normal and Dehydrated Brain: A Research Study and Review

BackgroundIn hypertonic dehydration, fluid loss is known to be from the intracellular compartment. Also fluid loss in vascular bed increases the osmotic pressure and results with decrease of fluid flow to these compartments. We applied hypertonic dehydration by fluid restriction to evaluate the effects of dehydration on posttraumatic brain edema and swelling. MethodsFour groups, each including 10 pigs composed the study group. Group 1: we performed craniectomy and followed up for 1 hour for intracranial pressure (ICP) measurement. Group 2: we performed craniectomy and hypertonic dehydration by fluid restriction for 3 days and followed up for 1 hour for ICP measurement. Group 3: we performed brain trauma with Madsen trauma model. Group 4: we performed brain trauma after dehydration. FindingsICP was found to be 10 mm Hg at the first, 6.5 mm Hg at the second group but increased gradually after trauma in group 3. At the fourth group ICP was high at the first 30 minutes. After the experiment brain fluid level was 80.9% at the first, 77.9% at the second, 83.5% at the third, and 82.8% at the fourth groups. Ultrastructural examination revealed normal brain tissue at the first, light degeneration at the second, advanced degeneration at the third group. The fourth group was also highly degenerated but was found to be better than the third group. Brain superoxide dismutase and malondialdehyde levels were found to be increased. InterpretationEven though hypertonic dehydration causes minimal cellular degeneration it increases the resistance of brain to trauma and high ICP especially at the first 30 minutes.