Ebru Dündar Yenilmez

Ultrastructural evaluation of the effect of N-acetylcysteine on methotrexate nephrotoxicity in rats.

The aim of this study is to investigate the possible protective effect of N-Acetylcysteine (NAC) against the likely methotrexate (MTX) toxicity on the kidney using ultrastructural together with biochemical data. Moreover, the immunohistochemical detection of Ki67 nuclear antigen is to be evaluated. Fifteen male Wistar albino rats, weighing 240-290 g, were divided into three equal groups: Rats receiving MTX alone, rats receiving MTX plus NAC treatment, and rats comprising the control group. MTX (18 mg/kg/day, body weight) in dissolved physiologic saline was administered intraperitoneally to rats during 3 days. For the MTX plus NAC group, N-Acetylcysteine (300 mg/kg/day, body weight) was administered together with MTX. At the end of the third day, all the rats were killed with cervical dislocation to obtain blood and tissue samples. Application of MTX principally induced prominent large vacuolization in the proximal convoluted tubule cells, and focal thickening in the glomerular basal lamina of some glomeruli. A decrease in tissue SOD (superoxide dismutase) and GSH-Px (glutathione peroxidase), and an increase in serum urea nitrogen and creatinine and in tissue MDA (malondialdehyde) levels were also seen in the MTX group. These changes were significantly reversed in the MTX-plus-NAC-treated group. Most of the vacuoles in the proximal convoluted tubule cells disappeared. Furthermore, an increase in antioxidant enzyme activities, a decrease in serum urea nitrogen and creatinine, and tissue MDA levels were all significant. Additionally, an increase in the number of Ki67 positive-stained cells in proximal tubules was also noted. In conclusion, NAC may be a promising substance against MTX-induced renal damage. It might be useful to use NAC supplementally to minimize MTX-induced nephrotoxicity.

Noninvasive prenatal diagnosis experience in the Çukurova Region of Southern Turkey: detecting paternal mutations of sickle cell anemia and β‐thalassemia in cell‐free fetal DNA using high‐resolution melting analysis

This study used a high‐resolution melting (HRM) technique to detect paternal mutations for the noninvasive prenatal diagnosis (NIPD) of β‐thalassemia and sickle cell anemia (HbS). We also determined the levels of cell‐free fetal DNA and total cell‐free DNA.

New Perspectives in Prenatal Diagnosis of Sickle Cell Anemia

Hemoglobin disorders such as thalassemias and sickle cell anemias can be avoided by detecting carriers, ensuring genetic counseling and prenatal diagnosis. Nowadays Chorionic villus sampling (CVS amniocentesis, and cordocentesis are still the most widely used invasive sampling methods for prenatal diagnosis of the fetus. These traditional methods are associated with a risk of fetal loss. The revelation of cell-free fetal DNA (cffDNA) in maternal plasma and serum provides the opportunity of noninvasive prenatal diagnosis (NIPD). Different encouraging clinical applications have arose such as noninvasive identification of fetal sexing, fetal Rhesus D, and the determination of the paternal alleles in maternal plasma. The determination of the presence or absence of paternally inherited alleles in maternal plasma of sickle cell disease (SCD) and β-thalassemia would allow the diagnosis of autosomal dominant diseases or the exclusion of autosomal recessive diseases of the fetuses, respectively. prenatal diagnosis of genetic diseases. Analysis of cffDNA in maternal plasma for NIPD has the advantage of being safer versus the invasive methods. Different technologies were used since the discovery of cffDNA for NIPD—especially high-resolution melting (HRM) analysis is one of those methods. Genotyping can be done with HRM without using labeled probes and more complex regions can be analyzed with unlabeled hybridization probes. High-resolution melting is a rapid and useful method to detect paternal alleles for the NIPD of SCD and thalassemias when the fetus has a risk for double heterozygote.

Effects of vitamin D on ovary in DHEA-treated PCOS rat model: A light and electron microscopic study

ABSTRACT Aim: The aim of this study was to investigate the effects of vitamin D treatment on ovary in experimentally designed polycystic ovary syndrome of female rats using light and electron microscopic techniques. Methods: Twenty-four female pre-pubertal rats were divided into control, DHEA and DHEA+Vit.D groups. In DHEA group, the PCOS rat model was developed by 6mg/kg/day dehydroepiandrosterone administration as subcutaneously injections. In DHEA+Vit.D group, 6 mg/kg/day DHEA and 120ng/100g/week 1,25(OH)2D3 was performed simultaneously. Controls were injected with vehicle alone. At the end of the 28 days, blood samples were collected and the ovarian tissues were taken for histological examinations. Results: FSH, LH levels, LH/FSH ratio, and testosterone levels showed a significant increase in DHEA group when compared with the control group. Moreover, these measurements were lower in the treatment group than the DHEA group. In DHEA group, increased number of atretic follicles and cystic follicles were seen with light microscopic analysis. Cystic follicles with attenuated granulosa cell layers and thickened theca cell layers and lipid accumulation in interstitial cells were observed by electron microscope. It is observed that atretic and cystic follicles were decreased as a result of vitamin D treatment. Conclusion: Our results indicate the curative role of vitamin D treatment on the androgen excess in PCOS rat model which causes abnormalities in ovarian morphology and functions. Vitamin D has positive effects on the hormonal and structural changes observed in PCOS, but it has been concluded that long-term use may be more beneficial.

A new biosensor for noninvasive determination of fetal RHD status in maternal blood of RhD negative pregnant women

Prenatal detection of the fetal RHD status can be useful in the management of RhD incompatibility to identify fetuses at risk of hemolytic disease. Hemolytic disease causes morbidity and mortality of the fetus in the neonatal period. The routine use of antenatal and postnatal anti-D prophylaxis has reduced the incidence of hemolytic disease of the fetus and newborn. This study describe the detection of fetal RhD antigens in blood of RhD negative pregnant women using a nanopolymer coated electrochemical biosensor for medical diagnosis. Cell free fetal DNA in maternal plasma was also used to genotyping fetal RHD status using multiplex real-time PCR. Twenty-six RhD negative pregnant women in different gestational ages were included in the study. RhD positive fetal antibodies detected with a developed biosensor in maternal blood of RhD negative mothers. The electrochemical measurements were performed on a PalmSens potentiostat, and corundum ceramic based screen printed gold electrode combined with the reference Ag/AgCl electrode, and the auxiliary Au/Pd (98/2%) electrode. Fetal RHD genotyping performed using fluorescence-based multiplex real-time PCR exons 5 and 7 of the RHD gene. The fetal RHD status of 26 RhD negative cases were detected 21 as RhD positive and 5 as RhD negative with electrochemical biosensor. Fetal RHD status confirmed with extracted fetal DNA in maternal plasma using multiplex real-time PCR RHD genotyping and by serological test after delivery. The new method for fetal RhD detection in early pregnancy is useful and can be carry out rapidly in clinical diagnosis. Using automated biosensors are reproducible, quick and results can be generated within a few minutes compared to noninvasive fetal RHD genotyping from maternal plasma with real-time PCR-based techniques. We suggest the biosensor techniques could become an alternative part of fetal RHD genotyping from maternal plasma as a prenatal screening in the management of RhD incompatibility.

Beneficial effects of rolipram, a phosphodiesterase 4 specific inhibitor, on testicular torsion-detorsion injury in rats

INTRODUCTION The aim of the study is to investigate the effect of Rolipram, a selective phosphodiesterase 4 inhibitor, on testicular torsion - detorsion injury. METHODS Sixty young male rats were divided into five groups. In each group, the right testes of six rats were removed four hours after detorsion for biochemical analysis, and the right testes of the remaining six rats were removed 24 h after detorsion for pathological analysis. In group 1 (sham-operated) right orchiectomy was performed without torsion, and right testes were sent to the laboratory for biochemical and pathologic analyses. In group 2 (control) torsion was applied to the right testes for 60 min, and detorsion was performed without the administration of Rolipram. In group 3 torsion was applied to the right testes for 60 min. 1 mg/kg Rolipram was administered 30 min before detorsion. In group 4 torsion was applied to the right testes for 60 min, and 1 mg/kg Rolipram was administered during detorsion. In group 5 torsion was applied to the right testes for 60 min. 1 mg/kg Rolipram was administered 30 min after detorsion. The malondialdehyde and nitric oxide levels were determined. The rates of necrosis and apoptosis were evaluated by histopathological examination. RESULTS The level of malondialdehyde was higher in the torsioned groups (Group 2, 3, 4, 5) than that in group 1 (p = 0.004). There was no statistically significant difference between the groups regarding the level of nitric oxide (p = 0.182). Apoptosis was higher in groups 2, 3 and 4 than in group 1; however, apoptosis was similar in group 1 and group 5 (p = 0.122). The level of necrosis in group 1 was similar to that in groups 4 and 5 (p = 0.194 and p = 0.847, respectively). CONCLUSION We suggest that the administration of Rolipram can decrease the rate of necrosis and apoptosis in testicular ischaemia-reperfusion injury.

Characterization CYP1A2, CYP2C9, CYP2C19 and CYP2D6 Polymorphisms Using HRMA in Psychiatry Patients with Schizophrenia and Bipolar Disease for Personalized Medicine

BACKGROUND The interindividual genetic variations in drug metabolizing enzymes effects the impact and toxicity in plenty of drugs. OBJECTIVE CYP1A2, CYP2C9, CYP2C19 and CYP2D6 gene polymorphisms were characterized using high resolution melting analysis (HRMA) in follow-up patients in psychiatry clinic as a preliminary preparation for personalized medicine. METHOD Genotyping of CYP1A2*1F, CYP2C9 *2, *3, CYP2C19 *2, *3 and *17 and CYP2D6 *3, *4 was conducted in 101 patients using HRMA. Genotype and allele frequencies of the CYP variants were found to be in equilibrium with the Hardy-Weinberg equation. RESULTS The frequency of the CYP1A2*1F allele in schizophrenia and bipolar disease was 0.694 and 0.255, respectively. The CYP2C9 allele frequencies were 0.087 (CYP2C9*2), and 0.549 (CYP2C9*3) for bipolar; 0.278 (CYP2C9*2) and 0.648 (CYP2C9*3) in schizophrenias. The CYP2C19*2 and *17 allele frequencies was 0.111 and 0.185 in schizophrenia and variant *2 was 0.117 and variant *17 was 0.255 in bipolar group. The frequency of the CYP2D6*3 allele was 0.027 in schizophrenias. The frequencies for the CYP2D6*4 variant were 0.092 and 0.096 in schizophrenia and bipolar groups, respectively. CONCLUSION The knowledge in pharmacogenomic and also the developments in molecular genetics are growing rapidly. In future, this can be expected to provide new methodologies in the prediction of the activity in drug metabolizing enzymes. The HRMA is a rapid and useful technique to identify the genotypes for drug dosage adjustment before therapy in psychiatry patients.

The effects of fetal hemoglobin levels on sickle cell anemia patients bone biochemistry / Fetal hemoglobin seviyesinin orak hücre anemisi hastalarının kemik biyokimyası üzerine etkisi

Abstract Objective: The aim of this study was to evaluate the possible roles of fetal hemoglobin levels on bone parameters in sickle cell anemia (SCA) patients. Methods: Blood samples taken from 56 SS and 47 control totally 103 subjects were included in this research work according to their fetal hemoglobin levels. Fetal hemoglobin, bone mineral density, bone specific alkaline phosphatase, calcium, osteocalcin, 25-OH vitamin D and hematological parameters were measured and analyzed in the study. Results: Statistical analysis showed that, lower bone mineral density and biochemical bone parameters significantly correlated with low fetal hemoglobin levels at SCA patients. Conclusion: It was concluded that fetal hemoglobin level is a good index for bone status in sickle cell anemia patients. Özet Amaç: Bu calışmanın amacı fetal hemoglobin seviyelerinin, orak hücre anemisi hastalarının kemik parametreleri üzerine olası etkisinin değerlendirilmesidir. Metod: 56’sı SS, 47’si kontrol grubu olmak üzere 103 kan örneği, fetal hemoglobin düzeylerine göre çalışmaya dahil edildi. Çalışmada fetal hemoglobin, kemik mineral yoğunluğu, kemik spesifik alkalen fosfataz, kalsiyum, osteokalsin, 25-OH vitamin D ve hematolojik parametreler analiz edildi. Bulgular: İstatistiksel analizler, duşuk kemik mineral yoğunluğu ve biyokimyasal kemik parametrelerinin orak hücre anemisi hastalarında fetal hemoglobin seviyeleri ile yakından ilişkili olduğunu göstermiştir. Sonuç: Orak hücre anemisi hastalarının kemik durumunu için fetal hemoglobin seviyesinin iyi bir parametre olduğu belirlenmiştir.

İnsan Lökosit Antijenleri, Yapı ve İşlevleri

Doku Uygunluk Antijenleri Kompleksi (MHC) insanda, Insan Lokosit Antijenleri (HLA) adini alir. Dort ana gruba ayrilan bu bolgede MHC Sinif I (HLA-A, -B, -C, -E, -F, -G), MHC Sinif II (HLA-DR, -DP, -DQ, -DO, -DN), MHC Sinif III (C2, C4A, C4B, PF, TNF-α, -β) ve MHC Sinif IV (SK12W, Hsp70, AIF-I, IC7, B144, LTB, TNF, LTA, IkBL, BATI, MICA, MICB) antijenleri yer almaktadir. Ayni zamanda MHC Sinif IV bolgesi, inflamatuar bolge olarak adlandirilmaktadir. HLA molekullerinin temel gorevi antijeni T lenfositlerine sunmak ve spesifik immun cevabi baslatmaktir. Sinif I HLA molekulleri CD8+ sitotoksik T hucrelere, Sinif II HLA molekulleri ise CD4+ yardimci T hucrelerine antijen sunarlar. Sinif III HLA molekulleri antijen sunumunda gorev almaz. Sinif III ve IV HLA molekulleri asil olarak enflamasyonda ve otoimmun hastaliklarda onemlidirler. HLA genleri, genomdaki en polimorfik genlerdir. En onemli kullanim yeri doku ve organ transplantasyonlarinda, doku uygunlugunun arastirilmasidir. Son zamanlarda uzerinde en cok calisilan konu HLA antijenlerinin hastaliklarla olan baglantisidir. Belirli HLA tipleri bazi hastaliklarda yuksek siklikta gorulmektedir. Bu derlemede HLA molekullerinin yapisi, islevleri ve klinikte kullanim alanlari hakkinda bilgi verilmistir. Ayni zamanda HLA polimorfizmleri ve bu antijenlerin hastalikla olan baglantisi hakkinda ozet bilgilerin sunulmasi amaclanmistir.