Browsing by Author "Altintas, N"

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    A pilot comparative study between serological and genetic investigations in relationship to clinical outcomes on patients with cystic echinococcosis
    Altintas, N; Karamil, SA; Turkum, O; Akil, M; Sakarya, A; Bozkaya, H; Soltys, J; Altintas, N
    The aim of this study was to investigate whether Enzyme-Linked Immunosorbent Assays (ELISA) and Western Blotting (WB) methods could contribute to the assessment of clinical outcomes in genotype-defined cystic echinococcosis (CE) patients. Twenty-nine human isolates and blood samples have been taken from patients who underwent surgery or percutaneous aspiration (PAIR) for therapeutic purposes at Ege University and Manisa Celal Bayar University Hospitals. All sera of patients were screened for the presence of E. granulosus IgG antibodies using in-house approved ELISA and WB methods. According to the ELISA results, five patients had high, thirteen patients had medium and eight patients had low specific antibody level response which ranged 1/640 -1/5000. Despite confirmed WB positivity three patients were found to be negative by ELISA. Immunoblot analysis of EgAg showed many protein bands with size of 8, 12, 20, 22, 24, 36, 75 and 90 kDa. Among of them, 8 - 12 kDa bands (90 %), 20 - 22 kDa and 36 kDa bands presented strong reactivity against human serum specimens. No serum samples from healthy control reacted with EgAg. Phylogenetic analysis of resulting COX1 and NAD1sequences has revealed that all patients in our study were infected with the E. granulosus G1-G3 genotype. There was no consistent correlation between results of ELISA and WB, the number or size of cysts and genotype. Our study brings a unique contribution in terms of relationship between serological investigation, disease genotypes and clinical outcomes.
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    Molecular Analysis of Cattle Isolates of Echinococcus granulosus in Manisa Province of Turkey
    Altintas, N; Oztatlici, M; Altintas, N; Unver, A; Sakarya, A
    Echinococcus granulosus is the causative agent of cystic echinococcosis (CE) in humans and many domestic animals, and still one of the most important global health problem in the world and in Turkey. Infection with metacestode causes severe illness and high economic losses. Several strains of Echinococcus have been identified based on the epidemiological and biological characteristics of strains. In this study, a total of 18 individual hydatid cyst samples from cattle were examined. They were obtained from central slaughterhouse in the province of Manisa/Turkey between 2010-2012. The total genomic DNA (gDNA) was extracted using RTA-DNA Isolation Kit (Gebze/Kocaeli, Turkey) according to manufacturer instructions from protoscoleces and cystic germinal membranes. The aim of this study was to provide molecular characterization of E. granulosus isolates which were obtained from cattles by using polymerase chain reaction (PCR) in Manisa province of Turkey. After PCR, to investigate the genetic characteristics of isolates, deoxyribonucleic acid sequencing of the mitochondrial cytochrome c oxidase subunit 1 (CO1) and nicotinamide adenine dinucleotide dehydrogenase subunit 1 (NAD1) genes were performed with ABI Prism Genetic Analyzer 3100 instrument. As a result of our study, all (18) cattle isolates were detected as E. granulosus sensu stricto (G1-G3 complex). This is the first molecular study report genotyping of Echinococcus isolates from cattle in Manisa province.
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    Genetical and Histological Investigation of Turkish Siblings With Spina Bifida Occulta Who Had Neurosurgical Intervention
    Altintas, N; Umur, S; Vatansever, S; Temiz, C; Selçuki, M; Selçuki, D; Örenay, S; Arslan, E
    Spina bifida is the one of the most frequently occurring birth defects. More children have spina bifida than muscular dystrophy, multiple sclerosis, and cystic fibrosis combined. Occulta present type is a mild very common form of spina bifida. The reported frequency of occurrence of spina bifida occulta (SBO) varies widely, depending largely on the age groups included in a particular study. The most accurate estimate of occurrence rate is 17% of examined spines. In present study we have examined and compared two Turkish siblings with spina bifida occulta who had neurosurgical intervention. Clinical, histopathological and cytogenetical analyses had been performed on mother and both siblings (brother and sister) diagnosed with spina bifida occulta. Hypertrichosis on their low-backs was diagnosed in both siblings. Magnetic resonance imaging (MRI) studies revealed low conus medullaris and thick filum terminale in brother and sister. The brother somatosensory evoked potential (SEP) results showed lumbar conduction blockade which was not found in sister. Despite the brothers's thick and fatty filum terminale sisters's seemed to be normal. Filum terminale sections from both siblings had normal appearance but atypical structure with increased amount of connective tissue and hyalinization areas. Sections from 9 years old boy were also prominent in glial cells. Genetical analysis revealed normal caryotype in 13 years old sister (46,XX), however deletions on chromosome 17 have been detected in 9 year-old brother [46,XY/46,XY,del(17)(q25)/47,+mar] and their mother [46,XX/46, XX,del(17)(q25)]. Our results show on strong correlation between the deletion of chromosome 17(q25) with genetical and histological results in both siblings with SBO. This is the first report of chromosome 17 (q25) deletion related to the SBO and its genetic connection with neural tube defects.
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    Role of the PON polymorphisms on progression of chronic hepatitis and cirrhosis
    Aldirmaz, M; Altintas, N; Var, A; Ellidokuz, E
    Aim and Background: The probable role of PON1-192, PON1-55, PON2-148, PON2-311 polymorphisms of the PON enzyme family, which are play a role in antioxidant pathways in the progression of chronic hepatitis to cirrhosis was investigated. Methods: The patient population included 64 chronic hepatitis patients without cirrhosis and 30 patients with cirrhosis which were diagnosed by biopsy compared to a control group (n=68) of normal healthy volunteers. All patients were recruited from the outpatient clinic of Gastroenterology, Celal Bayar University Faculty of Medicine. Genomic DNA of leukocytes was isolated by using a commercial isolation kit. PCR analysis was assessed for PON1-192, PON1-55, PON2-148, PON2-311 genotypes and the products digested with HinfI restriction enzyme to see the allelic polymorphisms. The PCR products were resolved on electrophoresis 2% agarose gel and visualized with a Syngene (USA) image analysis system. Odds ratios in 95% confidence interval were calculated for the two genotypes. For the statistical analyses SPSS version 10.0 was used. Results: The prevalence of PON1-192, PON1-55, PON2-148 genotypes were very different between the groups but was not statistically significant. The frequency of the SS and SC genotypes of PON2-311 in patients with hepatitis and cirrhosis was higher than the control group and their Odds Ratios were statistically significant. Conclusion: The prevalence of PON2 gene SS and SC genotypes of in patients was higher than that of healthy volunteers (ORs were 3,855 and 2,404 respectively). Similarly SS and SC genotypes of in patients with cirrhosis was higher than that of patients with hepatitis (ORs were 3,436 and 2,223 respectively) These results suggested that the SS and SC genotypes of PON2 gene might cause a susceptibility for developing of hepatitis and progression of this condition to cirrhosis. It can be also speculated that CC genotype may be protective for the progression of disease.
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    The relationship between paraoxanase gene Leu-Met (55) and Gln-Arg (192) polymorphisms and coronary artery disease
    Taskiran, P; Cam, SF; Sekuri, C; Tuzun, N; Alioglu, E; Altintas, N; Berdeli, A
    Objectives: Paraoxonase (PON1) is a high-density lipoprotein (HDL)-associated esterase that hydrolyses lipoperoxides. PON1 serves as a protective factor against oxidative modification of LDL, suggesting that it may play an important role in the prevention of atherosclerotic process. Research has focused on two polymorphisms: leucine (L allele) to methionine (M allele) substitution at codon 55, and glutamine (A allele) to arginine (B allele) substitution at codon 192. Study design: We examined amino acid changes at codon 55 and 192 in the PON1 gene by polymerase chain reaction and using restriction enzymes in 120 patients (92 men, 28 women; mean age 48.2 +/- 4.3 years) with premature coronary artery disease (CAD) and in 102 healthy subjects (80 men, 22 women; mean age 46.8 +/- 5.2 years) with no history of CAD and a normal electrocardiogram. Results: Distribution of genotypes in the patient and control groups at codon 55 was 6.7% and 4.9% for MM, 46.7% and 29.4% for LM, 46.7% and 65.7% for LL, respectively. The frequencies of genotypes at codon 192 were as follows: 4.2% and 2% for RR, 40% and 35.3% for QR, and 55.8% and 62.8% for QQ, respectively. While the frequency of PON1 55M allele was higher in the CAD group (0.3 vs. 0.2), PON1 192R allele frequency did not differ (0.2). There was a significant relationship between the PON1 M/L55 polymorphism and CAD (p=0.017), whereas the R/Q192 polymorphism was not associated with CAD (p=0.445). Conclusion: These data suggest that the PON1 M/L55 polymorphism shows a significant relationship with CAD and the Q/R192 polymorphism is not a major risk factor causing susceptibility to CAD in our population.
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    Comparison of serological and clinical findings in Turkish patients with cystic echinococcosis
    Yolasigmaz, A; Reiterová, K; Turk, M; Reyhan, E; Bozdag, AD; Karababa, AO; Altintas, N; Altintas, N
    Cystic echinococcosis (CE), caused by the cestode Echinococcus granulosus, is potentially dangerous for humans. The aim of this study was to examine serological and clinical findings regarding cysts localisation and individual responses in 54 patients with CE. The majority of patients in this study were females (63%) and the average age was 46.3 years. Most of the patients lived in rural areas or kept a dog (46%) for a long time. The most frequent symptoms were hypochondrial pain (48.9%), epigastrial discomfort (27.7%), vomiting (21.3%), minor cough (12.8%), urticaria (6.3%), weakness (4.3%), fever (2.1%), side- or backache (4.3%). However, 17% of the patients showed no symptoms. In every case, the ultrasound (USG) and/or computer tomography (CT) investigations were positive. In most cases (53.2% of the patients) a single cyst was found but 46.8% of the patients had multiple cyst formations (from 2 to 9 cysts) located in the liver. Sporadic lung, splenetic, mesenterial, tibial and cerebral localisations were also found. The patients were individually treated with albendazol (10 - 15 mg/kg) five days prior and six months after the surgical treatment. Serum samples were investigated by the serological techniques: IHAT, ELISA and Western blot using hydatid fluid antigen. In the patient sera, the specific antibody levels were mostly increased after surgery. Different results were obtained only in two patients. In the first case, seroconversion was delayed. In the other case all ELISA results were negative, however, the Western blot analysis and surgery proved the presence of CE. The results suggest that the different antibody response of patients depends on the individual immune response. Multiple localization and various stages of CE cysts demonstrate the necessity of a complex approach for the confirmation of a correct diagnosis.
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    Cystic echinococcosis in Turkey: genetic variability and first record of the pig strain (G7) in the country
    Snabel, V; Altintas, N; D'Amelio, S; Nakao, M; Romig, T; Yolasigmaz, A; Gunes, K; Turk, M; Busi, M; Hüttner, M; Sevcová, D; Ito, A; Altintas, N; Dubinsky, P
    A sample of 22 Echinococcus granulosus isolates collected from 12 sheep and ten humans from a focus of cystic echinococcosis in western Turkey was examined by DNA sequencing of four mitochondrial genes (cox1, atp6, nad1, rrnS). Results demonstrated the presence of two species of E. granulosus complex, E. granulosus sensu stricto and E. canadensis. Of E. granulosus sensu stricto, the G1 genotype (including three microvariants) was found in 17 isolates from humans and sheep, the G3 genotype and an intermediate form G1/G3 in one isolate each (both from sheep). Of E. canadensis, the pig strain G7 was found in three isolates from sheep and human. This is the first report of this strain in Turkey. Its presence has implications for local control programs due to its shorter maturation rate in dogs compared with E. granulosus sensu stricto. Goat and/or wild boar are likely reservoirs for G7 in the region. We provided further data on the pattern and frequency of nucleotide substitutions within the G1/G3 cluster. Based on our results and GenBank records, G2 (Tasmanian sheep strain) is not considered as a discrete genotypic unit, as its sequences at polymorphic sites conform to microvariants of both G1 and (more often) G3.
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    Comparison of the multi-epitope recombinant antigen DIPOL and hydatid fluid for the diagnosis of patients with cystic echinococcosis
    Ozturk, EA; Manzano-Román, R; Sánchez-Ovejero, C; Caner, A; Angin, M; Gunduz, C; Karaman, Ü; Altintas, N; Bozkaya, H; Unalp, O; Dokumcu, Z; Divarci, E; Casulli, A; Altintas, N; Siles-Lucas, M; Unver, A
    The use of serological tests containing multiple immunodominant antigens rather than single antigens have the potential to improve the diagnostic performance in Cystic Echinococcoses (CE) as a complement tool to clear the inconclusive imaging data. Here, we comparatively evaluated the diagnostic value of Hydatid Fluid (HF) and the recently described recombinant multi-epitope antigen DIPOL in IgG-ELISA in a clinically defined cohort of CE patients. The serum samples from 149 CE patients were collected just before surgical or Percutaneous-Aspiration-Injection-Reaspiration (PAIR) procedures. Additionally, serum samples of patients with other parasitic infections (n=49) and healthy individuals (n=21) were also included in the study as controls. To investigate the association between the genotype of the parasite and DIPOL, cyst materials from 20 CE patients were sequenced. In terms of overall sensitivity, HF was higher than DIPOL (82.55%,78.52%, respectively). However, while the sensitivity of HF was higher than DIPOL in patients with active and transitional cysts (83.3%, 75.4%, respectively), sensitivity of DIPOL in inactive cysts was higher compared to HF (95.6%, 78.3%, respectively). The sensitivity of DIPOL depending on cyst stage was statistically significant (P=0.041). In terms of specificity, DIPOL was found to be better than HF (97.71%, 91.43%, respectively). By genotyping, the majority of 20 patients showed G1 genotype (80%). All patients harboring G3 and G1/G3 cyst genotypes were positive with both antigens, while 87.5% of patients with G1 genotype were seropositive with HF and 75% with DIPOL. The overall sensitivity and high specificity of DIPOL suggest that this recombinant protein containing immunodominant epitopes is a potential substitute for the HF by serological tests for the diagnosis of CE.
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