Ayfer Atalay

High Incidence of Hb D-Los Angeles [β121(GH4)Glu→Gln] in Denizli Province, Aegean Region of Turkey

Denizli Province is located in the inner part of the Aegean region of Turkey and is one of the target areas for premarital screening. Here we report the abnormal hemoglobins (Hbs) observed during a premarital screening program in our region. According to our results, Hb D-Los Angeles [β121(GH4)Glu→Gln (GAA→CAA] (also known as D-Punjab, D-North Carolina, D-Portugal, Oak Ridge and D-Chicago), is the most frequent abnormal Hb in this region.

Genetic Origin of Hb D-Los Angeles [β121(GH4)Glu→Gln, GAA→CAA] According to the β-Globin Gene Cluster Haplotypes

Hb D-Los Angeles (also known as D-Punjab, D-North Carolina, D-Portugal, D-Chicago and Oak Ridge) is an abnormal hemoglobin (Hb) with an amino acid substitution of glutamine for glutamic acid at codon 121 of the β-globin gene. The origin and spread of Hb D-Los Angeles is not known. This is due to lack of information and remains to be elucidated. According to published reports, the Hb D-Los Angeles mutation is mostly linked with Mediterranean haplotype I [+ − − − − + +]. Besides the Mediterranean haplotype, a novel haplotype was also reported from Thailand [− − + + − − + + +]. Here we report a new haplotype from Turkey [− + −− + + +] that has not been described before. These results suggest that the Hb D-Los Angeles mutation has at least three different genetic origins.

Angiotensin-Converting Enzyme I/D Polymorphism in Behçet’s Disease

Objective: To investigate a potential relationship between I/D polymorphism within intron 16 of the angiotensin-converting enzyme (ACE) gene located on human chromosome 17 and Behçet’s disease. Materials and Methods: Genomic DNA was obtained from 35 Turkish patients diagnosed with Behçet’s disease according to the International Study Group criteria and 150 healthy individuals. Polymerase chain reaction was used to detect the presence of I and D (insertion and deletion) alleles in intron 16 of the ACE gene in these DNA samples. Results: We found differences in ACE I/D polymorphism between Behçet’s disease and healthy controls (χ2 = 4.61, d.f. = 1, p = 0.044). In Behçet’s disease patients, the D allele frequency was 84.3% and I allele frequency 15.7%. Conclusion: An association between Behçet’s disease and ACE polymorphism may provide a useful basis for future molecular studies and therapeutic approaches in this complex disease.

HLA-B51 gene and its expression in association with Behçet’s Disease in Denizli Province of Turkey

Behçet’s Disease (BD) is a multisystemic inflammatory disorder as a triad of symptoms including recurrent oral and genital aphthous ulceration, and uveitis with unknown pathogenesis. Many researchers have tried to investigate the association of HLA-B51 gene with the BD. We aimed to investigate the association of the HLA-B51 gene and its expression, also polymorphic structure by PCR, RT-PCR and sequence specific oligonucleotide primers and probes in BD patients (n: 35) and control group (n: 50). According to our results, we did not observe any association in between HLA-B51 gene, its polymorphism, expression and BD patients.

Rare hemoglobin variant Hb Yaizu observed in Turkey.

Objective: To determine the characteristic features of the rare hemoglobin (Hb) variant Hb Yaizu to enable laboratory diagnosis of the hemoglobin variants during screening programs. Materials and Methods: Genomic DNA was obtained from the 4 members of a family living in Denizli province, an Aegean region of Turkey. Blood cell counts, hemoglobin composition, hemoglobin electrophoresis (both alkaline and acid), HPLC analysis, DNA sequencing and beta globin gene cluster haplotypes were done. Results: Hb Yaizu carriers were apparently healthy individuals. Hb Yaizu was slightly faster than Hb S at alkaline pH, but slower than Hb S at acidic pH in hemoglobin electrophoresis. An abnormal hemoglobin peak was observed with a retention time of 4.77 min in HPLC analysis attributed to Hb Yaizu. Two members of the family were heterozygous Hb Yaizu [beta 79(EF3) Asp>Asn] confirmed by DNA sequencing. The mutation was found to be linked with the Mediterranean haplotype I [+––––++]. Conclusion: We have presented the details of Hb Yaizu, a rare hemoglobin variant that may be important to hemoglobinopathy screening programs, although its clinical significance is unclear.

First observation of Hb Tunis (beta124(H2) Pro>Ser) in Turkey

Hb Tunis [beta124(H2)Pro>Ser] was reported from Tunisia in 1988. This hemoglobin variant was detected by isoelectric focusing moving just ahead of Hb A. It cannot be identified by standard hemoglobin electrophoresis due to its similar mobility to Hb A. It has normal stability and oxygen affinity and does not produce any clinical symptoms. Here, we report a heterozygous Hb Tunis [beta124(H2)Pro>Ser] case discovered for the first time in Turkey in a premarital screening program. This hemoglobin variant can be identified with high performance liquid chromatography analysis confirmed with DNA sequencing. We emphasize in our study the importance of an interdisciplinary collaborative study at the provincial basis for the success of the hemoglobinopathy control program.

The IL-8 Gene Polymorphisms in Behçet's Disease Observed in Denizli Province of Turkey.

ABSTRACT Behçet’s disease is a multisystemic inflammatory disorder as a triad of symptoms including recurrent oral and genital aphthous ulceration and uveitis with unknown pathogenesis. IL-8, a proinflammatory cytokine, has been found increased in the active stage of BD. DNA samples were obtained from 88 patients with BD and 112 healthy control subjects in Denizli province of Turkey. All genotyping experiments of SNPs in IL-8 gene were performed using polymerase chain reaction-restriction fragment polymorphism. We found that IL-8 −845 T > C and −738 T > A sites are non-polymorphic. There were no differences in the polymorphisms of IL-8 +396 G/T, +781 C/T, and +1633 C/T sites except IL-8 −251 T > A in between patients and healthy controls. Analysis of IL-8 polymorphisms indicates that the distribution of frequencies seems to be associated with −251 T > A and gender, −251 T > A and erythema nodosum, −251 T > A and ocular involvement, +781 C > T and erythema nodosum, +396 G > T and pathergy positivity, and +1633 C > T and papulopustular lesion. We demonstrated that the frequencies of IL-8 haplotypes were significantly different with BD patients than control group. We found that the distribution of IL-8 haplotypes was significantly different with genital ulcers, ocular involvement, positive pathergy test, erythema nodosum, papulopustular lesions, and arthritis with BD patients than healthy control individuals. Our study suggests that IL-8 gene polymorphisms may affect susceptibility to BD and increase the risk of developing disease. In order to confirm and assess the association of IL-8 and other cytokine gene polymorphisms in the pathophysiology of BD, large cohort studies are needed.

Hb Beograd [β121(GH4)Glu→Val, GAA→GTA] in the Turkish Population

Hb Beograd [β121(GH4)Glu→Val, GAA→GTA] is a rare variant first reported in Yugoslavia and then in Turkey, Australia and New Zealand. We report two further unrelated cases from Turkey. The importance of identifying Hb Beograd at the molecular level, especially in regions where Hb D-Los Angeles [β121(GH4)Glu→Gln, GAA→CAA] is prevalent, is emphasized.

Analysis of the population genetic structure of Hb D‐Los Angeles [β121 (GH4) Glu→Gln GAA→CAA] in Denizli, Turkey; genetic diversity, historical demography and estimation of the mutation rates based on haplotype variation

Understanding the genetic origin of the Hb D‐Los Angeles hemoglobin may elucidate population interactions such as movements, migrations, and environmental effects on mutation mechanisms in human biology throughout history. Our study aimed to understand the genetic origin of Hb D‐Los Angeles based on haplotype data, observed in the Denizli province of Turkey.

Molecular Studies on the Origin of the Hb G-Coushatta Mutation in Denizli Province of Turkey

Hb G-Coushatta [b22(B4)Glu?Ala] was reported for the first time by Schneider et al. (1964) in an American Indian family. This variant hemoglobin migrates like HbS at alkaline pH but shows no clinical effects, and it is observed in geographically separated ethnic groups (Li et al. 1999). Several researchers also reported Hb G-Coushatta from unrelated families residing in various regions of Turkey (Li et al. 1999; Dincol et al. 1989; Sozmen et al. 1990; Yenice et al. 2000; Atalay et al. 2005). DNA polymorphisms have been used to define the origin of common mutant alleles of the b-globin gene. According to Orkin et al. (1982) and Antonarakis et al. (1982), the microhaplotype polymorphism of the b-globin gene designated the ‘‘framework’’ (FW) is defined by five single nucleotide polymorphisms (SNPs). FW1 and FW2 are common worldwide; FW3 is frequent in Africans, Europeans, and West Asians. FW3A, an intermediate structure between FW2 and FW3, is characteristic for East Asian populations (Antonarakis et al. 1982). Regarding the Hb G-Coushatta mutation, Li et al. (1999) examined the haplotype of the b-globin gene cluster in the Coushatta Tribe of Louisiana and native Chinese Hb G-Coushatta carriers to determine the genetic origin of this variant; they found different genetic origins in association with the haplotypes and frameworks in the Louisiana and Chinese cases. In our study, we examined the haplotypes and frameworks of the Hb G-Coushatta carriers residing in Denizli Province of Turkey.