Screening of polymorphisms in the folate pathway in Turkish pediatric Acute Lymphoblastic Leukemia patients

Background and aim: Folate metabolic pathway plays a significant role in leukemogenesis because of its necessity for nucleotide synthesis and DNA methylation. Folate deficiency causes DNA damage. Thus polymorphisms of folate-related genes may affect the susceptibility to childhood Acute Lymphoblastic Leukemia (ALL). MTHFR (Methylenetetrahydrofolate Reductase), DHFR (Dihydrofolate reductase), CBS (Cystathionine b-synthase) and TYMS (Thymidylate Synthase) have an important role in folate pathway because their activated variants modulate synthesis of DNA and levels of folate. In this study, we aimed to investigate whether polymorphisms in genes related to folate metabolic pathway influence the risk to childhood ALL.Subject and methods: The patient groups who were diagnosed with childhood ALL at Losante Pediatric Hematology-Oncology Hospital and healthy control groups were included in the study. MTHFR 677 CT, MTHFR 1298 A-C, CBS 844ins68, DHFR 19-bp and TYMS 1494del6 polymorphisms were screened. Genotyping of these polymorphisms was performed by Restriction Fragment Length Polymorphism (RFLP) analysis and Real Time Polymerase chain Reaction (Real Time-PCR).Results: In total, we have screened 5 polymorphisms in the studied genes. The results were compared between childhood ALL patients and healthy groups. Genotype frequencies of MTHFR 677 C-T, MTHFR 1298 A-C, CBS 844ins68 and DHFR 19-bp del were similar for childhood ALL patients and healthy groups. However, statistical results showed that TYMS 1494del6 may be associated with ALL pathogenesis (p < 0.001).Conclusion: We showed that TYMS polymorphism (rs2853542) may be associated with ALL pathogenesis. In addition, our results demonstrated that MTHFR, DHFR and CBS do not affect development of leukemia. Our study displays also importance as it is the first screening results to identify association with the studied polymorphisms in Turkish patients with childhood ALL and determination of the frequency in Turkish population

Štetni učinci pušenja tijekom trudnoće na DNA i razine reaktivnih oblika kisika (ROS) u krvi majke i novorođenčeta

Some of the genotoxic/carcinogenic substances or metabolites in cigarette smoke are capable of passing through the placenta and harming a newborn’s health. Smoking is also known as a factor in the formation of oxidative damage and the main mechanism involved in the carcinogenic process. Predetermining this genotoxic risk can be successfully achieved by measuring certain parameters of oxidative stress. The comet assay is considered an important biomarker for the evaluation of genotoxic substances and is effective for detecting DNA damage caused by smoking. This study examined third trimester bloods and the cord blood of 28 actively smoking and 22 non-smoking mothers in terms of DNA damage and oxidative stress parameters. Cu/Zn superoxide dismutase (CuZn-SOD), malondialdehyde (MDA), catalase (CAT), plasma nitrite/nitrates (NO2•/NO3•), selenium-dependent glutathione peroxidase (Se-GPx), Cu, and Zn levels were measured as indicators of oxidative damage. There were no significant increases in DNA damage of the actively smoking pregnant group in comparison with the non-smoking pregnant group, either in the third trimester or cord blood. Oxidative stress parameters of smoker and non-smoker groups were statistically different for MDA (p<0.05), CuZn-SOD (p<0.01), Se-GPx (p<0.05) values while the difference was not significant for NO2•/NO3•, CAT, Zn, and Cu values. The same values were also investigated in cord blood, and only NO2•/NO3• (p<0.01), Se-GPx (p<0.01 and CAT (p<0.001) values were found statistically different. Smoking mothers may have been exposed to more oxidative stress than non-smoking mothers.Pojedine genotoksične/kancerogene supstancije ili metaboliti u cigaretnom dimu mogu proći kroz posteljicu i naštetiti zdravlju novorođenčeta. Pušenje je također poznato kao čimbenik pri nastanku oksidacijskog oštećenja DNA i u procesu kancerogeneze. Ovaj genotoksični rizik može se uspješno odrediti mjerenjem određenih parametara oksidacijskog stresa. Komet-test smatra se važnim biološkim biljegom pri evaluaciji genotoksičnih supstancija i iznimno učinkovitim sredstvom pronalaženja oštećenja DNA uzrokovanih pušenjem. Ova studija proučava krv trudnica u trećem tromjesečju trudnoće i fetalnu krv 28 majki aktivnih pušačica te 22-ju majki nepušačica vezano za oksidacijska oštećenja DNA i parametre oksidacijskog stresa. Razine Cu/Zn superoksidne dismutaze (CuZn-SOD), malondialdehida (MDA), katalaze (CAT), nitrita/nitrata u plazmi (NO2-/NO3-), selenijeve glutation peroksidaze (Se-GPx), Cu i Zn mjerene su kao pokazatelji oksidacijskog oštećenja. Nije bilo značajnih povećanja oštećenja DNA u skupini trudnica aktivnih pušačica u usporedbi sa skupinom trudnica nepušačica, ni u krvi iz trećeg tromjesečja ni u fetalnoj krvi. Parametri oksidacijskog stresa pušačke i nepušačke skupine bili su statistički različiti za vrijednosti MDA (p<0,05), CuZn-SOD (p<0,01), Se-GPx (P<0,05), dok razlika nije bila značajna za vrijednosti NO2-/NO3-, CAT, Zn i Cu. Iste su vrijednosti ispitane i u fetalnoj krvi, a jedino su vrijednosti NO2-/NO3- (p<0,01), Se-GPx (p<0,01) i CAT (p<0,001) bile statistički različite. Vjerojatno je da su majke pušačice bile izložene većem oksidacijskom stresu od majki nepušačica

DNA methyltransferase expression differs with proliferation in childhood acute lymphoblastic leukemia

Ezer, Ustun/0000-0002-1536-7898; KARABULUT, HALIL GURHAN/0000-0002-2842-0029WOS: 000279308400044PubMed: 19763880…The Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK)We acknowledge to Timur Tuncali, Ongun Onaran and Uygar Tazebay for their technical support and to Kenan Kose for performing statistical analysis. This study was granted by The Scientific and Technological Research Council of Turkey (TUBITAK)

Mycoplasma pneumoniae Infection Associated With Pancytopenia A Case Report

Immune hemolytic anemia is a rare condition in childhood. Cold agglutinins have been implicated in the etiology of the hemolysis and frequently observed during Mycoplasma pneumoniae infections. We present here a case of cold agglutinin-related hemolytic anemia, thrombocytopenia, and leukopenia secondary to M. pneumoniae associated pneumonia. It is suggested that even though very rare, M. pneumoniae infection should be considered as the underlying disease in a patient presenting with pancytopenia

Detection of TET2, KRAS and CBL variants by Next Generation Sequencing and analysis of their correlation with JAK2 and FLT3 in childhood AML

Background: Acute myeloid leukemia (AML) is a heterogeneous clonal disorder in terms of cytogenetic and molecular aberrations. Ten-Eleven-Translocation 2 (TET2), Kirsten rat sarcoma viral oncogene homolog (KRAS), and Casitas B-cell lymphoma (CBL) have an important role pathogenesis of acute myeloid leukemia (AML) and their activated mutations confer proliferative and survival signals. Aim: In this study, we aimed to find possible genetic markers for molecular analysis in childhood AML by screening hot-spot exons of TET2, KRAS, and CBL using Next Generation Sequencing (NGS) analysis. In addition, association between found variants and mutations of Januse Kinase-2 (JAK2) and Fms-Related Tyrosine Kinase (FLT3) were analyzed which are important prognostic risk factors for AML. Methods: Eight patients who were diagnosed with pediatric AML at Losante Pediatric Hematology–Oncology Hospital were included to the study. Hot-spot exons of TET2, KRAS and CBL genes were screened using the NGS method. Furthermore, FLT3-Internal Tandem Duplicate (FLT3-ITD) and JAK2-V617F were analyzed by Real Time Polymerase chain Reaction (Real Time-PCR). Results: In total, we identified 20 variants in studied genes by NGS. In our patient group, 16 variants in the TET2 (seven novel, seven missense and two silent), two variants in the KRAS (one missense and one intronic) and two variants in the CBL (two novel) were found. All of AML patients were found negative for JAK V617 F. Three of the eight patients (37.5%) showed mutations of both FLT3-ITD and TET2, KRAS, CBL. Conclusion: We found novel mutations for TET2, KRAS, and CBL. The detected variants in this article seem to be the first screening results of genes studied by NGS in childhood AML patients. Our results also showed some degree of association between FLT3-ITD and TET2, KRAS, CBL mutations

Malondialdehyde, Antioxidant Enzymes, and Renal Tubular Functions in Children with Iron Deficiency or Iron-Deficiency Anemia

We aimed to investigate the effects of iron deficiency (ID) or iron-deficiency anemia (IDA) on oxidative stress and renal tubular functions before and after treatment of children. A total of 30 children with a diagnosis of IDA constituted the IDA group and 32 children with a diagnosis of ID constituted the ID group. Control group consisted 38 age-matched children. Serum ferritin, soluble transferrin receptor (sTfR), serum, and urinary sodium (Na), potassium (K), calcium (Ca), phosphorus (P), creatinine (Cr), uric acid (UA), urinary N-acetyl-beta-d-glucosaminidase (NAG) levels, and intra-erythrocyte malondialdehyde (MDA), catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) levels were measured before and after iron therapy in the IDA and ID groups, whereas it was studied once in the control group. We have divided the study group in groups according to age (infants < 2 years, children 3-9 years, and adolescents 10-15 years). Patients with IDA (infant, adolescent) and ID (infant, children, and adolescent) had a significantly high level of MDA in post-treatment period in comparison to those of healthy control. Patients with IDA (children, adolescent) and ID (infant, children) had a significantly high level of pre-treatment GSH-Px than controls. Post-treatment SOD was lower in IDA (children and adolescent) groups than control and post-treatment CAT was lower in IDA and ID (adolescent) groups than control. These findings show that ferrous sulfate used in the treatment of ID or IDA could lead to oxidative stress; however, a marked deterioration of in proximal renal tubular functions was not seen

HLA-matched family hematopoetic stem cell transplantation in children with beta thalassemia major: The experience of the Turkish Pediatric Bone Marrow Transplantation Group

WOS: 000310804300021PubMed ID: 22931438From January 1991 to June 2009, 245 children with beta thalassemia major who underwent their first allogeneic HSCT in Turkey and who were followed for a minimum of one yr post-transplantation were enrolled this study. The median age of the patients was 6.6 yr old (range, 122 yr). The distribution of Pesaro risk class I, II, and III categories was 41, 130, and 63 children, respectively. The median serum ferritin level was 2203 ng/mL. Eighty-eight patients received bone marrow (BM) stem cells; 137, peripheral blood (PB) stem cells; and 20, cord blood (CB) stem cells. The donors were HLA-matched siblings or parents. Median engraftment times were shorter in PBSCT patients compared with the BMT group (p < 0.001). Grade II-IV acute GvHD was observed in 33 children (13.5%), while cGvHD was observed in 28 patients (12.5%), eight of whom had the extensive form. Thalassemic reconstitution was observed in 43 (17%) of the transplant patients. Post-transplant aplasia occurred in three patients, and the TRM rate was 7.75%. Seventeen patients were lost after 100 days. The thalassemia-free survival and OS rates were 68% (95% CI, 61.874.2) and 85.0% (95% CI, 80.289.8), respectively. We believe that this study is important because it is the first multicenter national data for children with beta thalassemia major receiving HSCT

G-CSF-mobilized haploidentical peripheral blood stem cell transplantation in children with poor prognostic nonmalignant disorders

Haploidentical hematopoletic stem cell transplantation (HSCT) is currently one of the alternative curative treatment options for some nonmalignant but also for malignant diseases. However, concerns regarding its safety cause delays in time and a successful outcome. Between 2000 and 2005, twenty-one children with poor prognostic nonmalignant disorders, 13 boys and 8 girls, with a median age of 12 months, underwent 28 haploidentical peripheral HSCT. Immunomagnetic bead depletion device (CliniMACS) was used for indirect T-cell depletion. Indications for transplant were severe combined immunodeficiency (n = 16), osteopetrosis (n = 2), MDS (n = 1), amegakaryocytic thrombocytopenia (n = 1), and aplastic anemia (n = 1). Five patients (24%) had lung infection at the time of transplantation. The patients received a median of 25.67 x 10(6) G-CSF-mobilized peripheral CD34(+) progenitor cells and a median of 4.19 x 10(4) T-lymphocytes per kilogram of body weight with a T-cell depletion rate of median 4.59 logs. The rate of total engraftment was 66.6%. Median times for leukocyte and platelet engraftment were 14 and 16 days, respectively. The 6-year projected survival was 32% for all patients and 29.76% for patients with severe combined immunodeficiency (SCID). The rates of transplant-related mortality, graft failure, and severe GvHD were 14.2, 33.4%, and 8.3%, respectively. Infection was the main cause of death. The poor outcome may be explained with the poor prognostic factors of our patients such as the type of SCID in most cases (T-B-SCID), the median age over 6 months and the presence of lung infection in some children at the time of transplantation

Determinaton of Depression, Anxiety and Hopelessness Situations at Parents whose Children Are Followed in Gulhane Military Medical Faculty, Pediatric Hematology and Oncology Clinics Due to Any Malignancy or Chronic Disease

Introduction: Chronic systemic diseases in childhood have negatively affecting the quality of life and debilitating effects for both children and parents. In our study, we investigated depression, anxiety and hopelessness situations at parents of children with these diseases. Materials and methods: The study was done at parents of children diagnosed with malignancy or chronic disease in GATA Department of Pediatrics Heath and Disease, Pediatric Hematology and Oncology Clinics. Beck Depression Scale, Beck Anxiety Scale and Beck Hopelessness Scale were applied to the participants. Results: Parents of children, who are followed due to malignancy or chronic disease in department of pediatrics heath and disease, pediatric hematology and oncology clinics, constituted the study group. 60 mothers and 51 fathers as study group and 64 mothers and 45 fathers as control group were enrolled in the study between 1st July 2009 and 1st June 2010. The mean age of the parents in study group was 35,7&#177;5,1 and 33,3 5,6 age in control group. The depression score was significantly higher statistically in study group (p=0,035). No difference was fond for the anxiety and hopelessness scores between the groups (p=0,064 and p=0,695 respectively). There was no difference for depression, hopelessness and anxiety scores between mothers and fathers of the children (p=0,217, p=0,447, p=0,102, respectively). Conclusion: Without gender discrimination the parents of children with malignancy and chronic disease are in the risk group for depression. It is necessary to support the parents both socially and psychologically. [TAF Prev Med Bull 2012; 11(5.000): 577-582