Background/aim: Biotinidase deficiency (BTD) is characterized by broad genotypic variants and uns... more Background/aim: Biotinidase deficiency (BTD) is characterized by broad genotypic variants and unsteady biotinidase activity. Increasing enzyme activity and maturation throughout childhood. This study aims to reveal the course of biotinidase activities in a long-term follow-up period and concordance with their genotypes .Participants/Methods: A total of 1,773 biotinidase enzyme (BT) activity measurements were performed in 711 newborns with variants in the BTD gene over a 4-year follow-up period. Biochemical phenotyping was classified into four groups based on the highest measured enzyme activity level during the follow-up: Profound(≤10%), Partial(10.1-30%), Heterozygous(30.1-66.5%), and Normal(>66.6%). Results: The number of participants with BTD in the biochemical phenotype groups assigned based on the first measurement was 59, 217, 314, and 121 in the profound, partial, heterozygous, and normal groups respectively. Based on the highest measurement value during follow-up, the num...
Although it is known that there is generally a good correlation between genotypes and phenotypes,... more Although it is known that there is generally a good correlation between genotypes and phenotypes, the number of studies reporting discrepancies has recently increased, exclusively between milder genotypes and their phenotypes due to the complex nature of the CYP21A2 gene and methodological pitfalls. This study aimed to assess CYP21A2 genotyping in children with 21-hydroxylase deficiency (21-OHD) and establish their predictive genotype–phenotype correlation features using a large cohort in Southeastern Anatolia’s ethnically diverse population. The patients were classified into three groups: salt-wasting (SW), simple virilizing (SV) and non-classical (NC). The genotypes were categorized into six groups due to residual enzyme activity: null–A–B–C–D–E. CYP21A2 genotyping was performed by sequence-specific primer and sequenced with next generation sequencing (NGS), and the expected phenotypes were compared to the observed phenotypes. A total of 118 unrelated children with 21-OHD were included in this study (61% SW, 24.5% SV and 14.5% NC). The pathogenic variants were found in 79.5% of 171 mutated alleles (60.2%, 22.2%, and 17.6% in SW, SV and NC, respectively). Patient distribution based on genotype groups was as follows: null—16.1%, A—41.4%, B—6.0%, C—14.4%, E—22%). In2G was the most common pathogenic variant (33.9% of all alleles) and the most common variant in the three phenotype groups (SW—38.8%, SV—22.2% and NC—23.3%). The total genotype–phenotype correlation was 81.5%. The correlations of the null and A groups were 100% and 76.1%, respectively, while it was lower in group B and poor in group C (71.4% and 23.5%, respectively). This study revealed that the concordance rates of the severe genotypes with their phenotypes were good, while those of the milder genotypes were poor. The discrepancies could have resulted from the complex characteristics of 21-OHD genotyping and the limitations of using NGS alone without integrating with other comprehensive methods.
Maturity‐onset diabetes of the young (MODY) is often misdiagnosed as other types of diabetes beca... more Maturity‐onset diabetes of the young (MODY) is often misdiagnosed as other types of diabetes because it is overlooked due to atypical clinical presentations. This study aims to reveal the clinical and laboratory clues and examine their compatibility with MODY genotypes.
Acute Myeloid Leukemia (AML) is a hematopoietic stem cell disorder. Many of the tumor development... more Acute Myeloid Leukemia (AML) is a hematopoietic stem cell disorder. Many of the tumor development steps are processes regulated by Ca2+ dependent signaling pathways found in tumor growth. Calcium channels, which can be blocked by pharmacological agents such as verapamil, are important in the regulation of intracellular Ca2+ homeostasis. The failure to stop cell cycle and to achieve programmed cell death are the most important factors in AML or other types of cancers and are considered to be the real cause of cancer. Our aim in this study was to investigate the effect of verapamil on apoptosis and cell cycle in AML cell line, HL-60, using flow cytometry. The effective dose of verapamil was determined as 50 mg / ml by MTT method, and was applied to HL-60 cells, subsequently, they were assessed for apoptosis and cell cycle. After 50 mg / ml of verapamil application, apoptotic cell ratio was found to be 77.7% after 24 hours (p< 0.05). In cell cycle analysis, the ratio of diploid cell...
ABSTRACT Purpose: Pterygium, one of the most common ocular surface diseases, is characterized by ... more ABSTRACT Purpose: Pterygium, one of the most common ocular surface diseases, is characterized by inflammatory infiltrates, proliferation, angiogenesis, fibrosis, and extracellular matrix breakdown. The objective of this study was to elucidate the levels of the intercellular adhesion molecule (ICAM)-2, and ICAM-3 gene and protein expressions in pterygium. Methods: A total of 59 patients with pterygium were included in this study. mRNA from pterygial and conjunctival autograft tissues were extracted, and real-time polymerase chain reaction on the BioMark HD dynamic array system was performed for the ICAM-2 and ICAM-3 gene expressions. ICAM-2 and ICAM-3 protein expressions using western blot and immunohistochemistry methods were also investigated in pterygial and conjunctival autograft tissues. Results: ICAM-2 and ICAM-3 gene expressions were markedly augmented in pterygial tissues (P = 0.0018 and P = 0.0023, respectively). Significant increases in protein expressions in pterygial tissues were also detected for ICAM-2 and ICAM-3 (P = 0.0116 and P = 0.0252, respectively). In the immunohistochemical studies, there was a marked increase in ICAM-3 (P = 0.0152), but not in ICAM-2 (P = 0.1041), protein expressions in pterygial tissues. Significant positive correlations between pterygia grading with ICAM-2 protein expression (P = 0.0398) and ICAM-3 immunohistochemical scores (P = 0.0138) were observed. Conclusion: These results demonstrate, for the first time, the expressions of ICAM-2 and ICAM-3 in the pterygium. These findings may help to understand the signal transduction mechanisms in the pterygium formation and provide a new therapy strategy for pterygium treatment.
Pterygium is a benign fibrovascular proliferation that develops from the conjunctiva and invades ... more Pterygium is a benign fibrovascular proliferation that develops from the conjunctiva and invades the cornea. The etiology of this disorder remains unclear. Current treatment of pterygium is surgical. The postoperative recurrence rate of pterygium is reported be high. To the best of our knowledge, these results are the first to demonstrate the contribution of proteins expressions of the small GTPase Rho proteins in patients with pterygium. Our data showed that leukocyte RhoA, RhoB, RhoD, and RhoE protein expressions were markedly elevated in primary pterygium. However, no significant modifications were noted in pterygial tissues.
Pterygium is one of the most common ocular surface diseases, and characterized by inflammatory in... more Pterygium is one of the most common ocular surface diseases, and characterized by inflammatory infiltrates, proliferation, fibrosis, angiogenesis, and extracellular matrix breakdown. We investigated the association of polymorphisms in the RHO genes RHOA, RHOB, RHOC, RHOD, and RND3 (RHOE). The results of this study demonstrate for the first time the association of RHO genes with the pterygium. We displayed that the RHO gene polymorphisms were significantly associated with pterygium in the Turkish population.
Arteriyel hipertansiyon, obezite, dislipidemi veya diabetes mellitus gibi kardiyovasküler hastalı... more Arteriyel hipertansiyon, obezite, dislipidemi veya diabetes mellitus gibi kardiyovasküler hastalık (CVD) risk faktöleri ve buna ek olarak, miyokard infarktüsü, koroner arter hastalığı veya inme, tekrarlayan düşüklerin de dahil olduğu CVD’ler en yaygın hastalıklardır ve tüm dünyadaki ana ölüm sebeplerini oluşturmaktadırlar. Bu çalışmaya 2010-2013 yılları arasında Türkiye’nin güneydoğu bölgesinde CVD paneli uygulanmış birbirinden bağımsız 4.709 hasta dahil edilmiştir. Metodolojik olarak, bu hastalardan elde edilen kan örneklerinden DNA izolasyonu gerçekleştirilmiştir. Mutasyon analizi, CVD panelinde bulunan altı yaygın mutasyonu (Faktör VG1691A, Protrombin G20210A, Faktör XIII V34L, MTHFR A1298C, MTHFR C677T ve PAI-1 -675 4G/5G) taramak amacıyla real-time PCR reaksiyonu (RT-PCR) metodu kullanılarak yapılmıştır. Bu mutasyonların sıklığı homozigot formda sırasıyla sırasıyla % 0,57, % 0,25, % 2,61, % 13,78, % 9,34 ve % 24,27 olarak tespit edildi. Benzer şekilde, bu mutasyonların heterozi...
Cardiovascular disease (CVD) risk factors, such as arterial hypertension, obesity, dyslipidemia o... more Cardiovascular disease (CVD) risk factors, such as arterial hypertension, obesity, dyslipidemia or diabetes mellitus, as well as CVDs, including myocardial infarction, coronary artery disease or stroke, are the most prevalent diseases and account for the major causes of death worldwide. In the present study, 4,709 unrelated patients subjected to CVD panel in south-east part of Turkey between the years 2010– 2013 were enrolled and DNA was isolated from the blood samples of these patients. Mutation analyses were conducted using the real-time polymerase chain reaction (RT-PCR) method to screen six common mutations (Factor V G1691A, PT G20210A, Factor XIII V34L, MTHFR A1298C and C677T and PAI-1 -675 4G/5G) found in CVD panel. The prevalence of these mutations were 0.57%, 0.25%, 2.61%, 13.78%, 9.34% and 24.27% in homozygous form, respectively. Similarly, the mutation percent of them in heterozygous form were 7.43%, 3.44%, 24.91%, 44.94%, 41.09% and 45.66%, respectively. No mutation was d...
Familial mediterranean fever (FMF) is an autosomal recessive autoinflammatory disorder (MIM# 2491... more Familial mediterranean fever (FMF) is an autosomal recessive autoinflammatory disorder (MIM# 249100), particularly common in populations of Mediterranean extraction. MEFV gene, responsible for FMF, encoding pyrin has recently been mapped to chromosome 16p13.3. In the present study, 3,341 unrelated patients with the suspicion of FMF in south-east part of Turkey between the years 2009 and 2013 were enrolled and genomic sequences of exon 2 and exon 10 of the MEFV gene were scanned for mutations by direct sequencing. We identified 43 different type of mutations and 9 of them were novel. DNA was amplified by PCR and subjected to direct sequencing for the detection of MEFV gene mutations. Among the 3,341 patients, 1,598 (47.8 %) were males and 1,743 (52.1 %) were females. The mutations were heterozygous in 806 (62.3 %), compound heterozygous in 188 (14.5 %), homozygous in 281 (21.8 %) and mutations had complex genotype in 17 (1.32 %) patients. No mutation was detected in 2,051 (61.4 %) patients. The most frequent mutations were M694V, E148Q, M680I(G/C) and V726A. We could not find any significant differences between the two common mutations according to the gender. Molecular diagnosis of MEFV is a useful tool in clinical practice, thus a future study relating to genotype/phenotype correlation of FMF in more and larger group in Turkish population involving the whole MEFV gene mutations is necessary.
Background/aim: Biotinidase deficiency (BTD) is characterized by broad genotypic variants and uns... more Background/aim: Biotinidase deficiency (BTD) is characterized by broad genotypic variants and unsteady biotinidase activity. Increasing enzyme activity and maturation throughout childhood. This study aims to reveal the course of biotinidase activities in a long-term follow-up period and concordance with their genotypes .Participants/Methods: A total of 1,773 biotinidase enzyme (BT) activity measurements were performed in 711 newborns with variants in the BTD gene over a 4-year follow-up period. Biochemical phenotyping was classified into four groups based on the highest measured enzyme activity level during the follow-up: Profound(≤10%), Partial(10.1-30%), Heterozygous(30.1-66.5%), and Normal(>66.6%). Results: The number of participants with BTD in the biochemical phenotype groups assigned based on the first measurement was 59, 217, 314, and 121 in the profound, partial, heterozygous, and normal groups respectively. Based on the highest measurement value during follow-up, the num...
Although it is known that there is generally a good correlation between genotypes and phenotypes,... more Although it is known that there is generally a good correlation between genotypes and phenotypes, the number of studies reporting discrepancies has recently increased, exclusively between milder genotypes and their phenotypes due to the complex nature of the CYP21A2 gene and methodological pitfalls. This study aimed to assess CYP21A2 genotyping in children with 21-hydroxylase deficiency (21-OHD) and establish their predictive genotype–phenotype correlation features using a large cohort in Southeastern Anatolia’s ethnically diverse population. The patients were classified into three groups: salt-wasting (SW), simple virilizing (SV) and non-classical (NC). The genotypes were categorized into six groups due to residual enzyme activity: null–A–B–C–D–E. CYP21A2 genotyping was performed by sequence-specific primer and sequenced with next generation sequencing (NGS), and the expected phenotypes were compared to the observed phenotypes. A total of 118 unrelated children with 21-OHD were included in this study (61% SW, 24.5% SV and 14.5% NC). The pathogenic variants were found in 79.5% of 171 mutated alleles (60.2%, 22.2%, and 17.6% in SW, SV and NC, respectively). Patient distribution based on genotype groups was as follows: null—16.1%, A—41.4%, B—6.0%, C—14.4%, E—22%). In2G was the most common pathogenic variant (33.9% of all alleles) and the most common variant in the three phenotype groups (SW—38.8%, SV—22.2% and NC—23.3%). The total genotype–phenotype correlation was 81.5%. The correlations of the null and A groups were 100% and 76.1%, respectively, while it was lower in group B and poor in group C (71.4% and 23.5%, respectively). This study revealed that the concordance rates of the severe genotypes with their phenotypes were good, while those of the milder genotypes were poor. The discrepancies could have resulted from the complex characteristics of 21-OHD genotyping and the limitations of using NGS alone without integrating with other comprehensive methods.
Maturity‐onset diabetes of the young (MODY) is often misdiagnosed as other types of diabetes beca... more Maturity‐onset diabetes of the young (MODY) is often misdiagnosed as other types of diabetes because it is overlooked due to atypical clinical presentations. This study aims to reveal the clinical and laboratory clues and examine their compatibility with MODY genotypes.
Acute Myeloid Leukemia (AML) is a hematopoietic stem cell disorder. Many of the tumor development... more Acute Myeloid Leukemia (AML) is a hematopoietic stem cell disorder. Many of the tumor development steps are processes regulated by Ca2+ dependent signaling pathways found in tumor growth. Calcium channels, which can be blocked by pharmacological agents such as verapamil, are important in the regulation of intracellular Ca2+ homeostasis. The failure to stop cell cycle and to achieve programmed cell death are the most important factors in AML or other types of cancers and are considered to be the real cause of cancer. Our aim in this study was to investigate the effect of verapamil on apoptosis and cell cycle in AML cell line, HL-60, using flow cytometry. The effective dose of verapamil was determined as 50 mg / ml by MTT method, and was applied to HL-60 cells, subsequently, they were assessed for apoptosis and cell cycle. After 50 mg / ml of verapamil application, apoptotic cell ratio was found to be 77.7% after 24 hours (p< 0.05). In cell cycle analysis, the ratio of diploid cell...
ABSTRACT Purpose: Pterygium, one of the most common ocular surface diseases, is characterized by ... more ABSTRACT Purpose: Pterygium, one of the most common ocular surface diseases, is characterized by inflammatory infiltrates, proliferation, angiogenesis, fibrosis, and extracellular matrix breakdown. The objective of this study was to elucidate the levels of the intercellular adhesion molecule (ICAM)-2, and ICAM-3 gene and protein expressions in pterygium. Methods: A total of 59 patients with pterygium were included in this study. mRNA from pterygial and conjunctival autograft tissues were extracted, and real-time polymerase chain reaction on the BioMark HD dynamic array system was performed for the ICAM-2 and ICAM-3 gene expressions. ICAM-2 and ICAM-3 protein expressions using western blot and immunohistochemistry methods were also investigated in pterygial and conjunctival autograft tissues. Results: ICAM-2 and ICAM-3 gene expressions were markedly augmented in pterygial tissues (P = 0.0018 and P = 0.0023, respectively). Significant increases in protein expressions in pterygial tissues were also detected for ICAM-2 and ICAM-3 (P = 0.0116 and P = 0.0252, respectively). In the immunohistochemical studies, there was a marked increase in ICAM-3 (P = 0.0152), but not in ICAM-2 (P = 0.1041), protein expressions in pterygial tissues. Significant positive correlations between pterygia grading with ICAM-2 protein expression (P = 0.0398) and ICAM-3 immunohistochemical scores (P = 0.0138) were observed. Conclusion: These results demonstrate, for the first time, the expressions of ICAM-2 and ICAM-3 in the pterygium. These findings may help to understand the signal transduction mechanisms in the pterygium formation and provide a new therapy strategy for pterygium treatment.
Pterygium is a benign fibrovascular proliferation that develops from the conjunctiva and invades ... more Pterygium is a benign fibrovascular proliferation that develops from the conjunctiva and invades the cornea. The etiology of this disorder remains unclear. Current treatment of pterygium is surgical. The postoperative recurrence rate of pterygium is reported be high. To the best of our knowledge, these results are the first to demonstrate the contribution of proteins expressions of the small GTPase Rho proteins in patients with pterygium. Our data showed that leukocyte RhoA, RhoB, RhoD, and RhoE protein expressions were markedly elevated in primary pterygium. However, no significant modifications were noted in pterygial tissues.
Pterygium is one of the most common ocular surface diseases, and characterized by inflammatory in... more Pterygium is one of the most common ocular surface diseases, and characterized by inflammatory infiltrates, proliferation, fibrosis, angiogenesis, and extracellular matrix breakdown. We investigated the association of polymorphisms in the RHO genes RHOA, RHOB, RHOC, RHOD, and RND3 (RHOE). The results of this study demonstrate for the first time the association of RHO genes with the pterygium. We displayed that the RHO gene polymorphisms were significantly associated with pterygium in the Turkish population.
Arteriyel hipertansiyon, obezite, dislipidemi veya diabetes mellitus gibi kardiyovasküler hastalı... more Arteriyel hipertansiyon, obezite, dislipidemi veya diabetes mellitus gibi kardiyovasküler hastalık (CVD) risk faktöleri ve buna ek olarak, miyokard infarktüsü, koroner arter hastalığı veya inme, tekrarlayan düşüklerin de dahil olduğu CVD’ler en yaygın hastalıklardır ve tüm dünyadaki ana ölüm sebeplerini oluşturmaktadırlar. Bu çalışmaya 2010-2013 yılları arasında Türkiye’nin güneydoğu bölgesinde CVD paneli uygulanmış birbirinden bağımsız 4.709 hasta dahil edilmiştir. Metodolojik olarak, bu hastalardan elde edilen kan örneklerinden DNA izolasyonu gerçekleştirilmiştir. Mutasyon analizi, CVD panelinde bulunan altı yaygın mutasyonu (Faktör VG1691A, Protrombin G20210A, Faktör XIII V34L, MTHFR A1298C, MTHFR C677T ve PAI-1 -675 4G/5G) taramak amacıyla real-time PCR reaksiyonu (RT-PCR) metodu kullanılarak yapılmıştır. Bu mutasyonların sıklığı homozigot formda sırasıyla sırasıyla % 0,57, % 0,25, % 2,61, % 13,78, % 9,34 ve % 24,27 olarak tespit edildi. Benzer şekilde, bu mutasyonların heterozi...
Cardiovascular disease (CVD) risk factors, such as arterial hypertension, obesity, dyslipidemia o... more Cardiovascular disease (CVD) risk factors, such as arterial hypertension, obesity, dyslipidemia or diabetes mellitus, as well as CVDs, including myocardial infarction, coronary artery disease or stroke, are the most prevalent diseases and account for the major causes of death worldwide. In the present study, 4,709 unrelated patients subjected to CVD panel in south-east part of Turkey between the years 2010– 2013 were enrolled and DNA was isolated from the blood samples of these patients. Mutation analyses were conducted using the real-time polymerase chain reaction (RT-PCR) method to screen six common mutations (Factor V G1691A, PT G20210A, Factor XIII V34L, MTHFR A1298C and C677T and PAI-1 -675 4G/5G) found in CVD panel. The prevalence of these mutations were 0.57%, 0.25%, 2.61%, 13.78%, 9.34% and 24.27% in homozygous form, respectively. Similarly, the mutation percent of them in heterozygous form were 7.43%, 3.44%, 24.91%, 44.94%, 41.09% and 45.66%, respectively. No mutation was d...
Familial mediterranean fever (FMF) is an autosomal recessive autoinflammatory disorder (MIM# 2491... more Familial mediterranean fever (FMF) is an autosomal recessive autoinflammatory disorder (MIM# 249100), particularly common in populations of Mediterranean extraction. MEFV gene, responsible for FMF, encoding pyrin has recently been mapped to chromosome 16p13.3. In the present study, 3,341 unrelated patients with the suspicion of FMF in south-east part of Turkey between the years 2009 and 2013 were enrolled and genomic sequences of exon 2 and exon 10 of the MEFV gene were scanned for mutations by direct sequencing. We identified 43 different type of mutations and 9 of them were novel. DNA was amplified by PCR and subjected to direct sequencing for the detection of MEFV gene mutations. Among the 3,341 patients, 1,598 (47.8 %) were males and 1,743 (52.1 %) were females. The mutations were heterozygous in 806 (62.3 %), compound heterozygous in 188 (14.5 %), homozygous in 281 (21.8 %) and mutations had complex genotype in 17 (1.32 %) patients. No mutation was detected in 2,051 (61.4 %) patients. The most frequent mutations were M694V, E148Q, M680I(G/C) and V726A. We could not find any significant differences between the two common mutations according to the gender. Molecular diagnosis of MEFV is a useful tool in clinical practice, thus a future study relating to genotype/phenotype correlation of FMF in more and larger group in Turkish population involving the whole MEFV gene mutations is necessary.
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Papers by Gülper Nacarkahya