As a bioengineer with expertise in cell treatments, I have been involved in the establishment of four distinct GMP facilities dedicated to the manufacturing of advanced therapeutic medical products. I am presently employed as the manager overseeing the operations of the Cell and Biological Products Manufacturing Center at Life Park Hospital in Cyprus.
Purpose Autism spectrum disorder (ASD) is a neurodevelopmental condition that affects patients' a... more Purpose Autism spectrum disorder (ASD) is a neurodevelopmental condition that affects patients' ability to communicate, engage with others, and behave in certain ways. Despite the existence of several therapy possibilities, an effective treatment for ASD has not yet been identified. Cell therapies have been becoming increasingly recognized in recent years as a potential therapeutic approach for the management of ASD. Different types of cellular products are transplanted using different delivery methods as part of cell therapy, which has the ability to regulate the immune system, demonstrate paracrine, neuro-regenerative, anti-inflammatory, and anti-oxidative stress effects, as well as transfer healthy mitochondria. We have compared the results and findings of completed cell therapy clinical trials for the treatment of ASD in this systematic review. Methods A total of 547 studies were identified, in which 11 studies were found to be eligible to be included in this review as they were completed cell therapy clinical trials or clinical applications with quantitative results for the treatment of ASD patients. Results This systematic review provides an overview of clinical trials conducted with different types of cell therapy strategies for the treatment of ASD and their potential mechanisms of action. The limitations and future possibilities for this field of study, as well as the safety and efficacy of cell treatments in ASD, were reviewed. Conclusion Overall, the evidence suggests that various cell therapy methods may offer a novel and effective treatment option for individuals with ASD, although further research is needed to fully understand the optimal treatment strategy and therapeutic potential.
Genetic variants contribute to the pathogenesis of bronchopulmonary dysplasia (BPD). The aim of t... more Genetic variants contribute to the pathogenesis of bronchopulmonary dysplasia (BPD). The aim of this study is to evaluate the association of 45 SNPs with BPD susceptibility in a Turkish premature infant cohort. Infants with gestational age <32 weeks were included. Patients were divided into BPD or no-BPD groups according to oxygen need at 28 days of life, and stratified according to the severity of BPD. We genotyped 45 SNPs, previously identified as BPD risk factors, in 192 infants. A total of eight SNPs were associated with BPD risk at allele level, two of which (rs4883955 on KLF12 and rs9953270 on CHST9) were also associated at the genotype level. Functional relationship maps suggested an interaction between five of these genes, converging on WNT5A, a member of the WNT pathway known to be implicated in BPD pathogenesis. Dysfunctional CHST9 and KLF12 variants may contribute to BPD pathogenesis through an interaction with WNT5A. We suggest investigating the role of SNPs on different genes which are in relation with the Wnt pathway in BPD pathogenesis. We identified eight SNPs as risk factors for BPD in this study. In-silico functional maps show an interaction of the genes harboring these SNPs with the WNT pathway, supporting its role in BPD pathogenesis. NCT03467828. It is known that genetic factors may contribute to the development of BPD in preterm infants. Further studies are required to identify specific genes that play a role in the BPD pathway to evaluate them as a target for therapeutic interventions. Our study shows an association of BPD predisposition with certain polymorphisms on MBL2, NFKBIA, CEP170, MAGI2, and VEGFA genes at allele level and polymorphisms on CHST9 and KLF12 genes at both allele and genotype level. In-silico functional mapping shows a functional relationship of these five genes with WNT5A, suggesting that Wnt pathway disruption may play a role in BPD pathogenesis. It is known that genetic factors may contribute to the development of BPD in preterm infants. Further studies are required to identify specific genes that play a role in the BPD pathway to evaluate them as a target for therapeutic interventions. Our study shows an association of BPD predisposition with certain polymorphisms on MBL2, NFKBIA, CEP170, MAGI2, and VEGFA genes at allele level and polymorphisms on CHST9 and KLF12 genes at both allele and genotype level. In-silico functional mapping shows a functional relationship of these five genes with WNT5A, suggesting that Wnt pathway disruption may play a role in BPD pathogenesis.
Tropical theileriosis is a disease caused by infection with an apicomplexan parasite, Theileria a... more Tropical theileriosis is a disease caused by infection with an apicomplexan parasite, Theileria annulata, and giving rise to huge economic losses. In recent years, parasite resistance has been reported against the most effective antitheilerial drug used for the treatment of this disease. This emphasizes the need for alternative methods of treatment. Enolase is a key glycolytic enzyme and can be selected as a macromolecular target of therapy of tropical theileriosis. In this study, an intron sequence present in T. annulata enolase gene was removed by PCR-directed mutagenesis, and the gene was first cloned into pGEM-T Easy vector and then subcloned into pLATE31 vector, and expressed in Escherichia coli cells. The enzyme was purified by affinity chromatography using Ni-NTA agarose column. Steady-state kinetic parameters of the enzyme were determined using GraFit 3.0. High quantities (~65 mg/l of culture) of pure recombinant T. annulata enolase have been obtained in a higly purified form (&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;95 %). Homodimer form of purified protein was determined from the molecular weights obtained from a single band on SDS-PAGE (48 kDa) and from size exclusion chromatography (93 kDa). Enzyme kinetic measurements using 2-PGA as substrate gave a specific activity of ~40 U/mg, K m: 106 μM, kcat: 37 s(-1), and k cat/K m: 3.5 × 10(5) M(-1) s(-1). These values have been determined for the first time from this parasite enzyme, and availability of large quantities of enolase enzyme will facilitate further kinetic and structural characterization toward design of new antitheilerial drugs.
Genetic variants contribute to the pathogenesis of bronchopulmonary dysplasia (BPD). The aim of t... more Genetic variants contribute to the pathogenesis of bronchopulmonary dysplasia (BPD). The aim of this study is to evaluate the association of 45 SNPs with BPD susceptibility in a Turkish premature infant cohort. METHODS: Infants with gestational age <32 weeks were included. Patients were divided into BPD or no-BPD groups according to oxygen need at 28 days of life, and stratified according to the severity of BPD. We genotyped 45 SNPs, previously identified as BPD risk factors, in 192 infants. RESULTS: A total of eight SNPs were associated with BPD risk at allele level, two of which (rs4883955 on KLF12 and rs9953270 on CHST9) were also associated at the genotype level. Functional relationship maps suggested an interaction between five of these genes, converging on WNT5A, a member of the WNT pathway known to be implicated in BPD pathogenesis. Dysfunctional CHST9 and KLF12 variants may contribute to BPD pathogenesis through an interaction with WNT5A. CONCLUSIONS: We suggest investigating the role of SNPs on different genes which are in relation with the Wnt pathway in BPD pathogenesis. We identified eight SNPs as risk factors for BPD in this study. In-silico functional maps show an interaction of the genes harboring these SNPs with the WNT pathway, supporting its role in BPD pathogenesis. TRIAL REGISTRATION: NCT03467828.
Tropical theileriosis is a disease caused by infection with an apicomplexan parasite, Theileria a... more Tropical theileriosis is a disease caused by infection with an apicomplexan parasite, Theileria annulata, and giving rise to huge economic losses. In recent years, parasite resistance has been reported against the most effective antitheilerial drug used for the treatment of this disease. This emphasizes the need for alternative methods of treatment. Enolase is a key glycolytic enzyme and can be selected as a macromolecular target of therapy of tropical theileriosis. In this study, an intron sequence present in T. annulata enolase gene was removed by PCR-directed mutagenesis, and the gene was first cloned into pGEM-T Easy vector and then subcloned into pLATE31 vector, and expressed in Escherichia coli cells. The enzyme was purified by affinity chromatography using Ni-NTA agarose column. Steady-state kinetic parameters of the enzyme were determined using GraFit 3.0. High quantities (~65 mg/l of culture) of pure recombinant T. annulata enolase have been obtained in a higly purified form (&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;95 %). Homodimer form of purified protein was determined from the molecular weights obtained from a single band on SDS-PAGE (48 kDa) and from size exclusion chromatography (93 kDa). Enzyme kinetic measurements using 2-PGA as substrate gave a specific activity of ~40 U/mg, K m: 106 μM, kcat: 37 s(-1), and k cat/K m: 3.5 × 10(5) M(-1) s(-1). These values have been determined for the first time from this parasite enzyme, and availability of large quantities of enolase enzyme will facilitate further kinetic and structural characterization toward design of new antitheilerial drugs.
Amac: Bu calismada Turkiye’de erken dogan bebeklerde gorulen BPD insidansi ile IL-18RAP uzerinde ... more Amac: Bu calismada Turkiye’de erken dogan bebeklerde gorulen BPD insidansi ile IL-18RAP uzerinde bulunan rs3771150 polimorfizmi arasindaki iliski arastirilmistir. Materyal ve Metod: Bu polimorfizmin alel ve genotip frekanslari multipleks reaksiyonlar ve MALDI-TOF kutle spektrometresi yontemi ile elde edilen genotipleme sonuclari kullanilarak analiz edilmistir. Buna ek olarak hastalardan elde edilen klinik ve demografik veriler de BPD insidansi ile iliskileri acisindan degerlendirilmistir. Bulgular: Turk populasyonu icin rs3771150 polimorfizminin yabanil genotipi GG olarak belirlenmistir (n=49 kontrol, n=46 BPD hastasi). Diger genotipler (AG ve AA) analiz edildiginde BPD insidansi ile anlamli bir iliski kurulamamistir (p>0.05). Sonuc: Erken dogan 192 Turk bebekte rs3771150 polimorfizmi ile BPD insidansi arasinda anlamli bir iliski bulunamamistir.
ABSTRACT
Introduction: Bronchopulmonary dysplasia (BPD) is one of the most encountered lung disea... more ABSTRACT Introduction: Bronchopulmonary dysplasia (BPD) is one of the most encountered lung diseases seen in preterm infants. Bronchopulmonary dysplasia disease is mostly seen in preterm infants born between the 23rd-28th weeks of gestation which is the late canalicular/early saccular phase of lung development. Lungs of infants born in this phase, stop developing and the number of alveoli stays lower than mature babies. Today, discontinuance of alveoli development, surfactant deficiency and prematurity of the chest wall caused by inflammation and especially prematurity are seen as the major etiological factors of BPD. Aim: rs3771150 polymorphism on IL-18RAP was chosen to be investigated in terms of determining a relationship with BPD incidence seen in preterm infants born in Turkey. Materials and Method: Allele and genotype frequencies of this polymorphism was analyzed from genotyping results obtained by multiplex reactions and MALDI-TOF mass spectrometry method. In addition, clinical and demographic data obtained from the patients were also inquired in this study to determine any relationship with BPD incidence. Results: GG genotype has been determined as the wild type genotype of rs3771150 polymorphism for the Turkish population (n=49 in control group, n=46 in BPD group). When other genotypes (AG and AA) were analyzed no significant relationship was found with BPD incidence (p>0.05). Conclusion: No significant relationship could be determined between rs3771150 polymorphism and BPD incidence in 192 preterm Turkish infants.
Keywords: Bronchopulmonary dysplasia; multiplex PCR; single nucleotide polymorphism; SNP genotyping.
Tropical theileriosis is a disease caused by infection with an apicomplexan parasite, Theileria a... more Tropical theileriosis is a disease caused by infection with an apicomplexan parasite, Theileria annulata, and giving rise to huge economic losses. In recent years, parasite resistance has been reported against the most effective antitheilerial drug used for the treatment of this disease. This emphasizes the need for alternative methods of treatment. Enolase is a key glycolytic enzyme and can be selected as a macromolecular target of therapy of tropical theileriosis. In this study, an intron sequence present in T. annulata enolase gene was removed by PCR-directed mutagenesis, and the gene was first cloned into pGEM-T Easy vector and then subcloned into pLATE31 vector, and expressed in Escherichia coli cells. The enzyme was purified by affinity chromatography using Ni-NTA agarose column. Steady-state kinetic parameters of the enzyme were determined using GraFit 3.0. High quantities (~65 mg/l of culture) of pure recombinant T. annulata enolase have been obtained in a higly purified form (&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;95 %). Homodimer form of purified protein was determined from the molecular weights obtained from a single band on SDS-PAGE (48 kDa) and from size exclusion chromatography (93 kDa). Enzyme kinetic measurements using 2-PGA as substrate gave a specific activity of ~40 U/mg, K m: 106 μM, kcat: 37 s(-1), and k cat/K m: 3.5 × 10(5) M(-1) s(-1). These values have been determined for the first time from this parasite enzyme, and availability of large quantities of enolase enzyme will facilitate further kinetic and structural characterization toward design of new antitheilerial drugs.
ABSTRACT Background and objectives: The objective of this study is to determine the prevalence of... more ABSTRACT Background and objectives: The objective of this study is to determine the prevalence of factor V Leiden (G1691A), prothrombin (G20210A) and MTHFR (C677T) gene mutations in 201 Turkish patients who were referred to our clinic with venous thrombosis complications such as deep venous thrombosis, ischemic complications, thromboembolism and coronary artery disease. Methods: After isolation of genomic DNA from peripheral blood samples, polymerase chain reaction (PCR) and restriction fragment length polymorphism techniques were used for analysis. Results: Among patients with venous thrombosis complications, allelic frequencies were 0.33, 0.17 and 0.04 for MTHFR (C677T), factor V Leiden (G1691A) and prothrombin (G20210A) mutations respectively. Conclusion: Homozygosity for the MTHFR C677T mutation and/or presence of at least one copy of the A allele of the Factor V Leiden G1691A mutation was found to be associated with increased incidence of venous thrombosis complications in patients (p&lt;0.01). The combined impact of these mutations on venous thrombosis should also be taken into consideration. In our study, prothrombin (G20210A) mutation was found not to be associated with venous thrombosis complications. We also found that the prevalence of factor V Leiden (G1691A), prothrombin (G20210A) and MTHFR (C677T) gene mutations in Turkish patients with venous thrombosis are comparable to results of other studies performed in Turkish and Caucasian populations. We did not observe any significant gender dependency for the factor V Leiden (G1691A), prothrombin (G20210A) and MTHFR (C677T) gene mutations in venous thrombosis complications.
Background and objectives: The objective of this study is to determine the prevalence of factor V... more Background and objectives: The objective of this study is to determine the prevalence of factor V Leiden (G1691A), prothrombin (G20210A) and MTHFR (C677T) gene mutations in 201 Turkish patients who were referred to our clinic with venous thrombosis complications such as deep venous thrombosis, ischemic complications, thromboembolism and coronary artery disease. Methods: After isolation of genomic DNA from peripheral blood samples, polymerase chain reaction (PCR) and restriction fragment length polymorphism techniques were used for analysis. Results: Among patients with venous thrombosis complications, allelic frequencies were 0.33, 0.17 and 0.04 for MTHFR (C677T), factor V Leiden (G1691A) and prothrombin (G20210A) mutations respectively. Conclusion: Homozygosity for the MTHFR C677T mutation and/or presence of at least one copy of the A allele of the Factor V Leiden G1691A mutation was found to be associated with increased incidence of venous thrombosis complications in patients (p<0.01). The combined impact of these mutations on venous thrombosis should also be taken into consideration. In our study, prothrombin (G20210A) mutation was found not to be associated with venous thrombosis complications. We also found that the prevalence of factor V Leiden (G1691A), prothrombin (G20210A) and MTHFR (C677T) gene mutations in Turkish patients with venous thrombosis are comparable to results of other studies performed in Turkish and Caucasian populations. We did not observe any significant gender dependency for the factor V Leiden (G1691A), prothrombin (G20210A) and MTHFR (C677T) gene mutations in venous thrombosis complications.
Purpose Autism spectrum disorder (ASD) is a neurodevelopmental condition that affects patients' a... more Purpose Autism spectrum disorder (ASD) is a neurodevelopmental condition that affects patients' ability to communicate, engage with others, and behave in certain ways. Despite the existence of several therapy possibilities, an effective treatment for ASD has not yet been identified. Cell therapies have been becoming increasingly recognized in recent years as a potential therapeutic approach for the management of ASD. Different types of cellular products are transplanted using different delivery methods as part of cell therapy, which has the ability to regulate the immune system, demonstrate paracrine, neuro-regenerative, anti-inflammatory, and anti-oxidative stress effects, as well as transfer healthy mitochondria. We have compared the results and findings of completed cell therapy clinical trials for the treatment of ASD in this systematic review. Methods A total of 547 studies were identified, in which 11 studies were found to be eligible to be included in this review as they were completed cell therapy clinical trials or clinical applications with quantitative results for the treatment of ASD patients. Results This systematic review provides an overview of clinical trials conducted with different types of cell therapy strategies for the treatment of ASD and their potential mechanisms of action. The limitations and future possibilities for this field of study, as well as the safety and efficacy of cell treatments in ASD, were reviewed. Conclusion Overall, the evidence suggests that various cell therapy methods may offer a novel and effective treatment option for individuals with ASD, although further research is needed to fully understand the optimal treatment strategy and therapeutic potential.
Genetic variants contribute to the pathogenesis of bronchopulmonary dysplasia (BPD). The aim of t... more Genetic variants contribute to the pathogenesis of bronchopulmonary dysplasia (BPD). The aim of this study is to evaluate the association of 45 SNPs with BPD susceptibility in a Turkish premature infant cohort. Infants with gestational age <32 weeks were included. Patients were divided into BPD or no-BPD groups according to oxygen need at 28 days of life, and stratified according to the severity of BPD. We genotyped 45 SNPs, previously identified as BPD risk factors, in 192 infants. A total of eight SNPs were associated with BPD risk at allele level, two of which (rs4883955 on KLF12 and rs9953270 on CHST9) were also associated at the genotype level. Functional relationship maps suggested an interaction between five of these genes, converging on WNT5A, a member of the WNT pathway known to be implicated in BPD pathogenesis. Dysfunctional CHST9 and KLF12 variants may contribute to BPD pathogenesis through an interaction with WNT5A. We suggest investigating the role of SNPs on different genes which are in relation with the Wnt pathway in BPD pathogenesis. We identified eight SNPs as risk factors for BPD in this study. In-silico functional maps show an interaction of the genes harboring these SNPs with the WNT pathway, supporting its role in BPD pathogenesis. NCT03467828. It is known that genetic factors may contribute to the development of BPD in preterm infants. Further studies are required to identify specific genes that play a role in the BPD pathway to evaluate them as a target for therapeutic interventions. Our study shows an association of BPD predisposition with certain polymorphisms on MBL2, NFKBIA, CEP170, MAGI2, and VEGFA genes at allele level and polymorphisms on CHST9 and KLF12 genes at both allele and genotype level. In-silico functional mapping shows a functional relationship of these five genes with WNT5A, suggesting that Wnt pathway disruption may play a role in BPD pathogenesis. It is known that genetic factors may contribute to the development of BPD in preterm infants. Further studies are required to identify specific genes that play a role in the BPD pathway to evaluate them as a target for therapeutic interventions. Our study shows an association of BPD predisposition with certain polymorphisms on MBL2, NFKBIA, CEP170, MAGI2, and VEGFA genes at allele level and polymorphisms on CHST9 and KLF12 genes at both allele and genotype level. In-silico functional mapping shows a functional relationship of these five genes with WNT5A, suggesting that Wnt pathway disruption may play a role in BPD pathogenesis.
Tropical theileriosis is a disease caused by infection with an apicomplexan parasite, Theileria a... more Tropical theileriosis is a disease caused by infection with an apicomplexan parasite, Theileria annulata, and giving rise to huge economic losses. In recent years, parasite resistance has been reported against the most effective antitheilerial drug used for the treatment of this disease. This emphasizes the need for alternative methods of treatment. Enolase is a key glycolytic enzyme and can be selected as a macromolecular target of therapy of tropical theileriosis. In this study, an intron sequence present in T. annulata enolase gene was removed by PCR-directed mutagenesis, and the gene was first cloned into pGEM-T Easy vector and then subcloned into pLATE31 vector, and expressed in Escherichia coli cells. The enzyme was purified by affinity chromatography using Ni-NTA agarose column. Steady-state kinetic parameters of the enzyme were determined using GraFit 3.0. High quantities (~65 mg/l of culture) of pure recombinant T. annulata enolase have been obtained in a higly purified form (&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;95 %). Homodimer form of purified protein was determined from the molecular weights obtained from a single band on SDS-PAGE (48 kDa) and from size exclusion chromatography (93 kDa). Enzyme kinetic measurements using 2-PGA as substrate gave a specific activity of ~40 U/mg, K m: 106 μM, kcat: 37 s(-1), and k cat/K m: 3.5 × 10(5) M(-1) s(-1). These values have been determined for the first time from this parasite enzyme, and availability of large quantities of enolase enzyme will facilitate further kinetic and structural characterization toward design of new antitheilerial drugs.
Genetic variants contribute to the pathogenesis of bronchopulmonary dysplasia (BPD). The aim of t... more Genetic variants contribute to the pathogenesis of bronchopulmonary dysplasia (BPD). The aim of this study is to evaluate the association of 45 SNPs with BPD susceptibility in a Turkish premature infant cohort. METHODS: Infants with gestational age <32 weeks were included. Patients were divided into BPD or no-BPD groups according to oxygen need at 28 days of life, and stratified according to the severity of BPD. We genotyped 45 SNPs, previously identified as BPD risk factors, in 192 infants. RESULTS: A total of eight SNPs were associated with BPD risk at allele level, two of which (rs4883955 on KLF12 and rs9953270 on CHST9) were also associated at the genotype level. Functional relationship maps suggested an interaction between five of these genes, converging on WNT5A, a member of the WNT pathway known to be implicated in BPD pathogenesis. Dysfunctional CHST9 and KLF12 variants may contribute to BPD pathogenesis through an interaction with WNT5A. CONCLUSIONS: We suggest investigating the role of SNPs on different genes which are in relation with the Wnt pathway in BPD pathogenesis. We identified eight SNPs as risk factors for BPD in this study. In-silico functional maps show an interaction of the genes harboring these SNPs with the WNT pathway, supporting its role in BPD pathogenesis. TRIAL REGISTRATION: NCT03467828.
Tropical theileriosis is a disease caused by infection with an apicomplexan parasite, Theileria a... more Tropical theileriosis is a disease caused by infection with an apicomplexan parasite, Theileria annulata, and giving rise to huge economic losses. In recent years, parasite resistance has been reported against the most effective antitheilerial drug used for the treatment of this disease. This emphasizes the need for alternative methods of treatment. Enolase is a key glycolytic enzyme and can be selected as a macromolecular target of therapy of tropical theileriosis. In this study, an intron sequence present in T. annulata enolase gene was removed by PCR-directed mutagenesis, and the gene was first cloned into pGEM-T Easy vector and then subcloned into pLATE31 vector, and expressed in Escherichia coli cells. The enzyme was purified by affinity chromatography using Ni-NTA agarose column. Steady-state kinetic parameters of the enzyme were determined using GraFit 3.0. High quantities (~65 mg/l of culture) of pure recombinant T. annulata enolase have been obtained in a higly purified form (&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;95 %). Homodimer form of purified protein was determined from the molecular weights obtained from a single band on SDS-PAGE (48 kDa) and from size exclusion chromatography (93 kDa). Enzyme kinetic measurements using 2-PGA as substrate gave a specific activity of ~40 U/mg, K m: 106 μM, kcat: 37 s(-1), and k cat/K m: 3.5 × 10(5) M(-1) s(-1). These values have been determined for the first time from this parasite enzyme, and availability of large quantities of enolase enzyme will facilitate further kinetic and structural characterization toward design of new antitheilerial drugs.
Amac: Bu calismada Turkiye’de erken dogan bebeklerde gorulen BPD insidansi ile IL-18RAP uzerinde ... more Amac: Bu calismada Turkiye’de erken dogan bebeklerde gorulen BPD insidansi ile IL-18RAP uzerinde bulunan rs3771150 polimorfizmi arasindaki iliski arastirilmistir. Materyal ve Metod: Bu polimorfizmin alel ve genotip frekanslari multipleks reaksiyonlar ve MALDI-TOF kutle spektrometresi yontemi ile elde edilen genotipleme sonuclari kullanilarak analiz edilmistir. Buna ek olarak hastalardan elde edilen klinik ve demografik veriler de BPD insidansi ile iliskileri acisindan degerlendirilmistir. Bulgular: Turk populasyonu icin rs3771150 polimorfizminin yabanil genotipi GG olarak belirlenmistir (n=49 kontrol, n=46 BPD hastasi). Diger genotipler (AG ve AA) analiz edildiginde BPD insidansi ile anlamli bir iliski kurulamamistir (p>0.05). Sonuc: Erken dogan 192 Turk bebekte rs3771150 polimorfizmi ile BPD insidansi arasinda anlamli bir iliski bulunamamistir.
ABSTRACT
Introduction: Bronchopulmonary dysplasia (BPD) is one of the most encountered lung disea... more ABSTRACT Introduction: Bronchopulmonary dysplasia (BPD) is one of the most encountered lung diseases seen in preterm infants. Bronchopulmonary dysplasia disease is mostly seen in preterm infants born between the 23rd-28th weeks of gestation which is the late canalicular/early saccular phase of lung development. Lungs of infants born in this phase, stop developing and the number of alveoli stays lower than mature babies. Today, discontinuance of alveoli development, surfactant deficiency and prematurity of the chest wall caused by inflammation and especially prematurity are seen as the major etiological factors of BPD. Aim: rs3771150 polymorphism on IL-18RAP was chosen to be investigated in terms of determining a relationship with BPD incidence seen in preterm infants born in Turkey. Materials and Method: Allele and genotype frequencies of this polymorphism was analyzed from genotyping results obtained by multiplex reactions and MALDI-TOF mass spectrometry method. In addition, clinical and demographic data obtained from the patients were also inquired in this study to determine any relationship with BPD incidence. Results: GG genotype has been determined as the wild type genotype of rs3771150 polymorphism for the Turkish population (n=49 in control group, n=46 in BPD group). When other genotypes (AG and AA) were analyzed no significant relationship was found with BPD incidence (p>0.05). Conclusion: No significant relationship could be determined between rs3771150 polymorphism and BPD incidence in 192 preterm Turkish infants.
Keywords: Bronchopulmonary dysplasia; multiplex PCR; single nucleotide polymorphism; SNP genotyping.
Tropical theileriosis is a disease caused by infection with an apicomplexan parasite, Theileria a... more Tropical theileriosis is a disease caused by infection with an apicomplexan parasite, Theileria annulata, and giving rise to huge economic losses. In recent years, parasite resistance has been reported against the most effective antitheilerial drug used for the treatment of this disease. This emphasizes the need for alternative methods of treatment. Enolase is a key glycolytic enzyme and can be selected as a macromolecular target of therapy of tropical theileriosis. In this study, an intron sequence present in T. annulata enolase gene was removed by PCR-directed mutagenesis, and the gene was first cloned into pGEM-T Easy vector and then subcloned into pLATE31 vector, and expressed in Escherichia coli cells. The enzyme was purified by affinity chromatography using Ni-NTA agarose column. Steady-state kinetic parameters of the enzyme were determined using GraFit 3.0. High quantities (~65 mg/l of culture) of pure recombinant T. annulata enolase have been obtained in a higly purified form (&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;95 %). Homodimer form of purified protein was determined from the molecular weights obtained from a single band on SDS-PAGE (48 kDa) and from size exclusion chromatography (93 kDa). Enzyme kinetic measurements using 2-PGA as substrate gave a specific activity of ~40 U/mg, K m: 106 μM, kcat: 37 s(-1), and k cat/K m: 3.5 × 10(5) M(-1) s(-1). These values have been determined for the first time from this parasite enzyme, and availability of large quantities of enolase enzyme will facilitate further kinetic and structural characterization toward design of new antitheilerial drugs.
ABSTRACT Background and objectives: The objective of this study is to determine the prevalence of... more ABSTRACT Background and objectives: The objective of this study is to determine the prevalence of factor V Leiden (G1691A), prothrombin (G20210A) and MTHFR (C677T) gene mutations in 201 Turkish patients who were referred to our clinic with venous thrombosis complications such as deep venous thrombosis, ischemic complications, thromboembolism and coronary artery disease. Methods: After isolation of genomic DNA from peripheral blood samples, polymerase chain reaction (PCR) and restriction fragment length polymorphism techniques were used for analysis. Results: Among patients with venous thrombosis complications, allelic frequencies were 0.33, 0.17 and 0.04 for MTHFR (C677T), factor V Leiden (G1691A) and prothrombin (G20210A) mutations respectively. Conclusion: Homozygosity for the MTHFR C677T mutation and/or presence of at least one copy of the A allele of the Factor V Leiden G1691A mutation was found to be associated with increased incidence of venous thrombosis complications in patients (p&lt;0.01). The combined impact of these mutations on venous thrombosis should also be taken into consideration. In our study, prothrombin (G20210A) mutation was found not to be associated with venous thrombosis complications. We also found that the prevalence of factor V Leiden (G1691A), prothrombin (G20210A) and MTHFR (C677T) gene mutations in Turkish patients with venous thrombosis are comparable to results of other studies performed in Turkish and Caucasian populations. We did not observe any significant gender dependency for the factor V Leiden (G1691A), prothrombin (G20210A) and MTHFR (C677T) gene mutations in venous thrombosis complications.
Background and objectives: The objective of this study is to determine the prevalence of factor V... more Background and objectives: The objective of this study is to determine the prevalence of factor V Leiden (G1691A), prothrombin (G20210A) and MTHFR (C677T) gene mutations in 201 Turkish patients who were referred to our clinic with venous thrombosis complications such as deep venous thrombosis, ischemic complications, thromboembolism and coronary artery disease. Methods: After isolation of genomic DNA from peripheral blood samples, polymerase chain reaction (PCR) and restriction fragment length polymorphism techniques were used for analysis. Results: Among patients with venous thrombosis complications, allelic frequencies were 0.33, 0.17 and 0.04 for MTHFR (C677T), factor V Leiden (G1691A) and prothrombin (G20210A) mutations respectively. Conclusion: Homozygosity for the MTHFR C677T mutation and/or presence of at least one copy of the A allele of the Factor V Leiden G1691A mutation was found to be associated with increased incidence of venous thrombosis complications in patients (p<0.01). The combined impact of these mutations on venous thrombosis should also be taken into consideration. In our study, prothrombin (G20210A) mutation was found not to be associated with venous thrombosis complications. We also found that the prevalence of factor V Leiden (G1691A), prothrombin (G20210A) and MTHFR (C677T) gene mutations in Turkish patients with venous thrombosis are comparable to results of other studies performed in Turkish and Caucasian populations. We did not observe any significant gender dependency for the factor V Leiden (G1691A), prothrombin (G20210A) and MTHFR (C677T) gene mutations in venous thrombosis complications.
Giriş Pek çok yetişkin dokuda bulunan mezenkimal kök hücreler (MKH), farklılaşmamış hücreler olma... more Giriş Pek çok yetişkin dokuda bulunan mezenkimal kök hücreler (MKH), farklılaşmamış hücreler olmaları, yüksek proliferasyon kapasiteleri ile kendini yenileyebilme yetenekleri ve mezodermal farklılaşma potansiyelleri ile hasarlı dokuların rejenerasyonunda kullanılmak için dikkat çekici kök hücre kaynaklarıdır (1). Günümüzde klinik olarak daha çok yetişkin kemik iliği (Kİ) ve adipoz doku (AD) kaynaklı hücreler kullanılıyor olsa da, bu dokuların toplanması için invazif prosedürler ve donör yaşı ile ilgili katı kurallar gerekmektedir. Ayrıca hastaların yaşı ilerledikçe, bu hastalardan elde edilen MKH'lerin klinik olarak etkinliği azalmaktadır. (2, 3). Bu sebeplerden dolayı alternatif MKH kaynağı doku arayışı, plasenta, umbilikal kord, kordon kanı ve amniyon sıvısı gibi primitif veya neo-natal dokulara doğru yönelmektedir (4-6). İnsan kordon kanında kök hücrelerin ve progenitör hücrelerin varlığı ilk kez 1985 yılında Dr. Hal Broxmeyer tarafından bulunmuştur. 1988 yılında ise kordon kanı hücreleri ilk defa başarılı bir şekilde Fransa'da Fanconi anemisi endikasyonlu üç hastaya transplante edilmiş ve başarılı sonuçlar alınmıştır (7). Etik konularla ilgili sorunlardan bağımsız olması, immüno-süpresif özellik göstererek allojenik kullanıma uygun olması ve invazif olmayan yöntemler ile kolayca izole edilebilmesi kordon kanı kaynaklı MKH'leri (KK-MKH) en uygun MKH kaynaklarından biri yapmaktadır
As well as possessing the ability to proliferate, differentiate and self-renewal properties in em... more As well as possessing the ability to proliferate, differentiate and self-renewal properties in embryonic development, stem cells can also provide regeneration in adult tissues. Besides regenerating cells in damaged tissue, they contribute to the protection of cells from apoptosis, stimulate stem cell potentials in a tissue, inhibit vasculogenesis and act as a prohibitory factor of inflammation by secreting signaling molecules (such as; cytokines, growth factors, anti-apoptotic factors, cell adhesion receptors). Besides providing supportive results, cellular therapies are also potential hopes to treat diseases which cannot be treated with conventional protocols under favour of these various mechanisms of stem cells.
While investigating the potential usage possibilities of stem cells for incurable diseases, an argument of defining cancer as a stem cell disease has emerged. Even after years of recovery following treatments with conventional chemotherapy and radiotherapy, relapses still can be diagnosed. This situations brings forward the idea that cancer might be a disease of stem cells. Recent studies investigating conventional chemotherapy and radiotherapy report that these conventional therapies are just targeting cells which are in a fast division cycle but not effecting the cells which are in a slower division cycle. For instance, sleeping cancer cells under the endosteum can awake and convert into more resistant cells with aggressive proliferation even after completing conventional treatment procedures.
Using nano-scaled cell-derived particles as extracellular vehicles, which are widely termed as microvesicles, exosomes or microparticles, have gathered attention in recent years. The main cause of interest on these well-known particles which are secreted by healthy cells is that stem cell derived exosomes have demonstrated great ability to provide therapeutically benefit and they also show potential as biomarkers for the cancer diagnosis.
This chapter will emphasize on one of the most important topics of medicine; stem cells and cancer stem cells. Furthermore, the importance of exosomes for early diagnosis of uro-oncological cancers will be discussed.
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Papers by Ayberk Akat
Introduction: Bronchopulmonary dysplasia (BPD) is one of the most encountered lung diseases seen in preterm infants. Bronchopulmonary dysplasia disease is mostly seen in preterm infants born between the 23rd-28th weeks of gestation which is the late canalicular/early saccular phase of lung development. Lungs of infants born in this phase, stop developing and the number of alveoli stays lower than mature babies. Today, discontinuance of alveoli development, surfactant deficiency and prematurity of the chest wall caused by inflammation and especially prematurity are seen as the major etiological factors of BPD.
Aim: rs3771150 polymorphism on IL-18RAP was chosen to be investigated in terms of determining a relationship with BPD incidence seen in preterm infants born in Turkey.
Materials and Method: Allele and genotype frequencies of this polymorphism was analyzed from genotyping results obtained by multiplex reactions and MALDI-TOF mass spectrometry method. In addition, clinical and demographic data obtained from the patients were also inquired in this study to determine any relationship with BPD incidence.
Results: GG genotype has been determined as the wild type genotype of rs3771150 polymorphism for the Turkish population (n=49 in control group, n=46 in BPD group). When other genotypes (AG and AA) were analyzed no significant relationship was found with BPD incidence (p>0.05).
Conclusion: No significant relationship could be determined between rs3771150 polymorphism and BPD incidence in 192 preterm Turkish infants.
Keywords: Bronchopulmonary dysplasia; multiplex PCR; single nucleotide polymorphism; SNP genotyping.
(G1691A), prothrombin (G20210A) and MTHFR (C677T) gene mutations in 201 Turkish patients who were referred
to our clinic with venous thrombosis complications such as deep venous thrombosis, ischemic complications,
thromboembolism and coronary artery disease. Methods: After isolation of genomic DNA from peripheral blood
samples, polymerase chain reaction (PCR) and restriction fragment length polymorphism techniques were used for
analysis.
Results: Among patients with venous thrombosis complications, allelic frequencies were 0.33, 0.17 and 0.04 for
MTHFR (C677T), factor V Leiden (G1691A) and prothrombin (G20210A) mutations respectively.
Conclusion: Homozygosity for the MTHFR C677T mutation and/or presence of at least one copy of the A allele
of the Factor V Leiden G1691A mutation was found to be associated with increased incidence of venous thrombosis
complications in patients (p<0.01). The combined impact of these mutations on venous thrombosis should also be
taken into consideration. In our study, prothrombin (G20210A) mutation was found not to be associated with venous
thrombosis complications. We also found that the prevalence of factor V Leiden (G1691A), prothrombin (G20210A)
and MTHFR (C677T) gene mutations in Turkish patients with venous thrombosis are comparable to results of other
studies performed in Turkish and Caucasian populations. We did not observe any significant gender dependency for
the factor V Leiden (G1691A), prothrombin (G20210A) and MTHFR (C677T) gene mutations in venous thrombosis
complications.
Introduction: Bronchopulmonary dysplasia (BPD) is one of the most encountered lung diseases seen in preterm infants. Bronchopulmonary dysplasia disease is mostly seen in preterm infants born between the 23rd-28th weeks of gestation which is the late canalicular/early saccular phase of lung development. Lungs of infants born in this phase, stop developing and the number of alveoli stays lower than mature babies. Today, discontinuance of alveoli development, surfactant deficiency and prematurity of the chest wall caused by inflammation and especially prematurity are seen as the major etiological factors of BPD.
Aim: rs3771150 polymorphism on IL-18RAP was chosen to be investigated in terms of determining a relationship with BPD incidence seen in preterm infants born in Turkey.
Materials and Method: Allele and genotype frequencies of this polymorphism was analyzed from genotyping results obtained by multiplex reactions and MALDI-TOF mass spectrometry method. In addition, clinical and demographic data obtained from the patients were also inquired in this study to determine any relationship with BPD incidence.
Results: GG genotype has been determined as the wild type genotype of rs3771150 polymorphism for the Turkish population (n=49 in control group, n=46 in BPD group). When other genotypes (AG and AA) were analyzed no significant relationship was found with BPD incidence (p>0.05).
Conclusion: No significant relationship could be determined between rs3771150 polymorphism and BPD incidence in 192 preterm Turkish infants.
Keywords: Bronchopulmonary dysplasia; multiplex PCR; single nucleotide polymorphism; SNP genotyping.
(G1691A), prothrombin (G20210A) and MTHFR (C677T) gene mutations in 201 Turkish patients who were referred
to our clinic with venous thrombosis complications such as deep venous thrombosis, ischemic complications,
thromboembolism and coronary artery disease. Methods: After isolation of genomic DNA from peripheral blood
samples, polymerase chain reaction (PCR) and restriction fragment length polymorphism techniques were used for
analysis.
Results: Among patients with venous thrombosis complications, allelic frequencies were 0.33, 0.17 and 0.04 for
MTHFR (C677T), factor V Leiden (G1691A) and prothrombin (G20210A) mutations respectively.
Conclusion: Homozygosity for the MTHFR C677T mutation and/or presence of at least one copy of the A allele
of the Factor V Leiden G1691A mutation was found to be associated with increased incidence of venous thrombosis
complications in patients (p<0.01). The combined impact of these mutations on venous thrombosis should also be
taken into consideration. In our study, prothrombin (G20210A) mutation was found not to be associated with venous
thrombosis complications. We also found that the prevalence of factor V Leiden (G1691A), prothrombin (G20210A)
and MTHFR (C677T) gene mutations in Turkish patients with venous thrombosis are comparable to results of other
studies performed in Turkish and Caucasian populations. We did not observe any significant gender dependency for
the factor V Leiden (G1691A), prothrombin (G20210A) and MTHFR (C677T) gene mutations in venous thrombosis
complications.
While investigating the potential usage possibilities of stem cells for incurable diseases, an argument of defining cancer as a stem cell disease has emerged. Even after years of recovery following treatments with conventional chemotherapy and radiotherapy, relapses still can be diagnosed. This situations brings forward the idea that cancer might be a disease of stem cells.
Recent studies investigating conventional chemotherapy and radiotherapy report that these conventional therapies are just targeting cells which are in a fast division cycle but not effecting the cells which are in a slower division cycle. For instance, sleeping cancer cells under the endosteum can awake and convert into more resistant cells with aggressive proliferation even after completing conventional treatment procedures.
Using nano-scaled cell-derived particles as extracellular vehicles, which are widely termed as microvesicles, exosomes or microparticles, have gathered attention in recent years. The main cause of interest on these well-known particles which are secreted by healthy cells is that stem cell derived exosomes have demonstrated great ability to provide therapeutically benefit and they also show potential as biomarkers for the cancer diagnosis.
This chapter will emphasize on one of the most important topics of medicine; stem cells and cancer stem cells. Furthermore, the importance of exosomes for early diagnosis of uro-oncological cancers will be discussed.
Keywords: stem cell, cancer stem cell, exosome.