Serap Yalin

Biomechanical evaluation in osteoporosis : ovariectomized rat model

The aim of our study was to investigate the effects of ovariectomy on rat femur biomechanical parameters. Bone mineral density (BMD) and histological investigation were also evaluated. Fourteen female Sprague–Dawley rats (seven ovariectomized, seven control) were used. BMD was measured by dual-energy X-ray absorbsiometry. Bone biomechanical parameters were measured in femoral midshaft with tensile test using a biomaterial testing machine and maximum load, stiffness, energy absorption capacity (structural properties), ultimate stress, ultimate strain, and elastic modulus (material properties) were calculated. Diaphyseal cortical bone thickness was measured by using histological method. The ovariectomized (OVX) rat femur’s BMD was 14% lower than control rats (p=0.006). Mean maximum load was 55% less than the control group’s (p=0.0001). Stiffness was 72% less in OVX rats (p=0.05). Femurs of rats with OVX had 32% less absorbed energy than controls (p=0.09). From the stress–strain curve ultimate stress, ultimate strain and elastic modulus was calculated. Elastic modulus was 53% less than controls (p=0.05). Ultimate stress decreased 21% in OVX rats (p=0.097). Ultimate strain was 25% less than controls in OVX rats. Cortical thickness was significantly decreased in OVX rats than in controls (p<0.05). In conclusion, femur biomechanical parameters are decreased in osteoporosis.

A study on the investigation of cadmium chloride genotoxicity in rat bone marrow using micronucleus test and chromosome aberration analysis

In this study, we investigated the genotoxic and cytotoxic potential of cadmium chloride (CdCl2)in Wistar rat tibia bone marrow cells, using the structural chromosomal aberration (SCA) and micronucleus (MN) test systems. CdCl2 was administered to adult female rats as repeated i.p. doses of 0.5 mg/kg b.w. for 18 week (four months) at 48 h intervals. Mitomycin C (MMC) was used as a positive control (2 mg/kg b.w.). This study shows that cadmium chloride treatment significantly induced the frequency of micronucleus in polychromatic erythrocytes in tibia bone marrow. This increase in micronucleus frequency shows that cadmium has a genotoxic effect on bone marrow at this level. Also, in order to determine cytotoxicity in bone marrow, the ratio of polychromatic erythrocytes to normochromatic erythrocytes was calculated in bone marrow. The results of this study indicate that CdCl2 decreased this ratio. The decrease of this ratio in bone marrow shows CdCl2 may lead to cytotoxicity. We have reported that 0.5 mg/kg-level chronic exposure to cadmium (Cd) has an injurious effect on bone marrow. Our findings indicate that CdCl2 has a cytotoxic and genotoxic effect on rat bone marrow at chronic exposure.

Rationale for an international consortium to study inherited genetic susceptibility to childhood acute lymphoblastic leukemia.

Acute lymphoblastic leukemia is the major pediatric cancer in developed countries. To date most association studies of acute lymphoblastic leukemia have been based on the candidate gene approach and have evaluated a restricted number of polymorphisms. Such studies have served to highlight difficulties in conducting statistically and methodologically rigorous investigations into acute lymphoblastic leukemia risk. Recent genome-wide association studies of childhood acute lymphoblastic leukemia have provided robust evidence that common variation at four genetic loci confers a modest increase in risk. The accumulated experience to date and relative lack of success of initial efforts to identify novel acute lymphoblastic leukemia predisposition loci emphasize the need for alternative study designs and methods. The International Childhood Acute Lymphoblastic Leukaemia Genetics Consortium includes 12 research groups in Europe, Asia, the Middle East and the Americas engaged in studying the genetics of acute lymphoblastic leukemia. The initial goal of this consortium is to identify and characterize low-penetrance susceptibility variants for acute lymphoblastic leukemia through association-based analyses. Efforts to develop genome-wide association studies of acute lymphoblastic leukemia, in terms of both sample size and single nucleotide polymorphism coverage, and to increase the number of single nucleotide polymorphisms taken forward to large-scale replication should lead to the identification of additional novel risk variants for acute lymphoblastic leukemia. Ethnic differences in the risk of acute lymphoblastic leukemia are well recognized and thus in assessing the interplay between inherited and non-genetic risk factors, analyses using different population cohorts with different incidence rates are likely to be highly informative. Given that the frequency of many acute lymphoblastic leukemia subgroups is small, identifying differential effects will realistically only be possible through multi-center pooled analyses. Here, we review the rationale for identifying genetic risk variants for acute lymphoblastic leukemia and our proposed strategy for establishing the International Childhood Acute Lymphoblastic Leukaemia Genetics Consortium.

Strontium ranelate treatment improves oxidative damage in osteoporotic rat model.

BACKGROUND Osteoporosis is the most common skeletal disorder and is considered a risk of fracture. Most medication used for the treatment of osteoporosis is antiresorptive; however, strontium ranelate (Sr) therapy in postmenopausal women has shown a double effect on resorption and bone formation. In this study, the effect of Sr on status of the oxidative stress and antioxidant defence system was investigated. METHODS Twenty-one adult albino female Wistar rats were used. The animals were randomly assigned into three groups, control (sham operated rats, received saline), OVX (ovariectomized rats), OVX + Sr (4 months later ovariectomy, strontium ranelate treatment was begun and continued for 120 days) each containing 7 animals. Strontium ranelate (500 mg/kg/day) and placebo (saline) were administered via oral gavage. At the end of the treatment, liver and kidney of rats were removed and malondialdehyde (MDA) level, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) activities were determined by biochemical analysis methods. RESULTS In liver, MDA levels were significantly higher in the OVX and OVX + Sr groups than the control group. GSH-Px activity decreased in OVX group and increased in OVX + Sr group compared with values of control group. CAT activity was increased in the OVX + Sr group when compared to control group. In kidney, MDA level was increased in OVX group. SOD activity was decreased in the OVX + Sr group. GSH-Px activity decreased in OVX group and increased in OVX + Sr group compared with control group. CAT activity increased in the OVX + Sr group when compared to control. CONCLUSION According to our results, Sr has preventive effect on oxidative damage in ovariectomized rats.

Exposure to gamma rays induces early alterations in skin in rodents: mechanical, biochemical and structural responses.

In this study, the effect of gamma rays has been investigated on the normal rat skin using biomechanical, biochemical and histological techniques. Seventeen male Wistar albino rats were divided into two groups (control (n=7) and irradiated (n=10)). The irradiated group was treated with a (60)Co gamma source at a dose of 10Gy at room temperature. Skin biomechanics were measured with tensile test using biomaterial testing machine and maximum load, stiffness, energy absorption capacity, ultimate stress, ultimate strain and elastic modulus were calculated. In the irradiated group, energy, strain and toughness were significantly lower than in the control group (p<0.05). However, strength, displacement, stiffness, stress and elastic modulus were similar to that of the control group (p>0.05). Catalase (CAT) activities and the levels of malondialdehyde (MDA) in the skin of rats were measured using the biochemical methods. MDA levels significantly increased whereas CAT activities decreased in the irradiated group as compared with the control group (p<0.05). Diameters of collagen fibers were measured by transmission electron microscopy. There was no significant difference (p>0.05) between control and irradiated groups for collagen fiber diameter. Thickness of epidermis was significantly lower than the control group. There were no changes in the epidermis between the irradiated group and the control group ultrastructurally. The results of this study show that the gamma irradiation has a significant effect on normal healthy skin.

Association of CYP2B6 G15631T polymorphism with acute leukemia susceptibility

The Cytochrome P450 (CYP) enzymes constitute one of the biggest gene families and play a vital role in the metabolism of endogenous biomolecules, drugs and xenobiotics. One of the members of this family, CYP2B6, plays a very important role in metabolizing carcinogens and medications. CYP2B6G15631T gene polymorphism reduces CYP2B6 enzyme activity. In this study, we aimed to determine whether any association exists between genetic polymorphism in CYP2B6G15631T and individual susceptibility to acute leukemia. Our study group consisted of 80 acute leukemia patients and 100 unrelated healthy volunteers as a control group. 44 of the acute leukemia patients were diagnosed with acute lymphoblastic leukemia (ALL) and 36 patients with acute myeloid leukemia (AML). Genomic DNA was isolated from peripheral blood and genomic DNA samples were assayed for restriction fragment length polymorphism in the CYP2B6 loci by PCR amplification followed by digestion with BsrI. The data were analyzed statistically employing chi-square and logistic regression analyses. The frequencies of GG genotype (wild type) were 40.9%, 50% and 67% in ALL, AML and control groups, respectively. The frequencies of polymorphic GT genotype (heterozygous variant) were found to be 59.1% in ALL patients, 50% in AML patients and 33% in controls. The TT genotype (homozygous variant) was not observed in either control or leukemia cases. Logistic regression analyses showed a significant correlation between the CYP2B6 G15631T polymorphism (GT) and acute leukemia patients (OR=2.481, 95% CI=1.353-4.551, p=0.003). Our findings indicate that GT genotype may be an important genetic determinant for acute leukemias. According to our knowledge, this is the first report of an association between acute leukemia cases and the CYP2B6 G15631T polymorphism.

Electrophysiological, biochemical and ultrastructural effects of radiotherapy on normal rat sciatic nerve.

Abstract Purpose: The aim of the present study was to evaluate the electrophysiological, biochemical and ultrastructural changes on the rat sciatic nerve after radiotherapy. Material and Methods: Thirty male Wistar albino rats were divided into three groups as: Control group (n = 10), Group I: 3 months after radiotherapy (n = 10), and Group II: 6 months after radiotherapy (n = 10). Groups I and II were irradiated with a 60Co gamma source. A dose of 20 Gy in 10 fractions was applied to Groups I and II. Compound motor action potentials (CMAP) were recorded in all groups. Superoxide dismutase (SOD) and catalase (CAT) activities and malondialdehyde (MDA) levels were measured in the sciatic nerve of rats using the biochemical methods. Ultrastructural changes were determined by electron microscopy. Results: In Groups I and II, the amplitude of CMAP was significantly lower and the latency was significantly higher than that of the control group. There were no significant differences between Groups I and II regarding the CMAP amplitude and latency. The MDA levels were significantly increased, whereas the SOD and CAT activities were significantly decreased in experimental groups when compared with the control group. However, there were no significant changes in these parameters between Groups I and II. Degeneration in myelinated nerve fibers was observed ultrastructurally only in the experimental groups. Significant changes were observed between the control group and experimental groups in terms of ultrastructural myelin grading score and axonal damage score. No significant differences were found between Groups I and II. Conclusions: These findings indicated that the dose of 20 Gy in 10 fractions radiotherapy caused neuropathic damages in normal rat sciatic nerve 3 and 6 months after irradiation.

N-Acetylcysteine-induced vasodilatation is modulated by KATP channels, Na+/K+-ATPase activity and intracellular calcium concentration: An in vitro study

BACKGROUND In this study, we aimed to investigate the role of ATP-sensitive potassium (KATP) channel, Na+/K+-ATPase activity, and intracellular calcium levels on the vasodilatory effect of N-acetylcysteine (NAC) in thoracic aorta by using electrophysiological and molecular techniques. METHODS Rat thoracic aorta ring preparations and cultured thoracic aorta cells were divided into four groups as control, 2mM NAC, 5mM NAC, and 10mM NAC. Thoracic aorta rings were isolated from rats for measurements of relaxation responses and Na+/K+-ATPase activity. In the cultured thoracic aorta cells, we measured the currents of KATP channel, the concentration of intracellular calcium and mRNA expression level of KATP channel subunits (KCNJ8, KCNJ11, ABCC8 and ABCC9). RESULTS The relaxation rate significantly increased in all NAC groups compared to control. Similarly, Na+/K+- ATPase activity also significantly decreased in NAC groups. Outward KATP channel current significantly increased in all NAC groups compared to the control group. Intracellular calcium concentration decreased significantly in all groups with compared control. mRNA expression level of ABCC8 subunit significantly increased in all NAC groups compared to the control group. Pearson correlation analysis showed that relaxation rate was significantly associated with KATP current, intracellular calcium concentration, Na+/K+-ATPase activity and mRNA expression level of ABCC8 subunit. CONCLUSION Our findings suggest that NAC relaxes vascular smooth muscle cells through a direct effect on KATP channels, by increasing outward K+ flux, partly by increasing mRNA expression of KATP subunit ABCC8, by decreasing in intracellular calcium and by decreasing in Na+/K+-ATPase activity.

IRS-2 G1057D polymorphism in Turkish patients with colorectal cancer

Introduction Gene polymorphisms have a broad range of analysis, but are of particular use in molecular medicine due to their potential in revealing the genetic tendency in diseases such as cancer, heart attack etc. These studies basically depend on mutations that can be detected by proper techniques. The genes coding the insulin receptor substrate (IRS) proteins are among the most widely analysed polymorphisms in various cancer types, in which a G1057D mutation is seen. Aim To determine the risk of colon cancer by analysing the IRS-2 gene polymorphism in Turkish patients. Material and methods A total of 161 newly diagnosed colorectal cancer patients were analysed and compared to 197 unrelated healthy controls. A polymerase chain reaction-based restriction fragment length polymorphism method was carried out. Results No differences were observed between the patient and control groups for both allele and genotype frequencies of the IRS-2 G1057D gene. Conclusions Our results demonstrated that IRS-2 G1057D polymorphism is not associated with colorectal cancer in the Turkish population. This research is a preliminary and original study in Turkish patients with colorectal cancer. It also provides population-level genetic data on IRS-2 in the Turkish population. Further studies should be performed on larger number of patients and controls for more reliable results about the genetic tendency in colorectal cancer in Turkey. The study is a collaborative work of different universities and scientists.

Is mycophenolate mofetil an alternative agent to corticosteroids in traumatic nerve paralysis

Objective The effects of an immunosuppressive agent, mycophenolate mofetil (MM), were investigated and compared with those of methylprednisolone (MP) and dexamethasone (DXM) on the traumatic nerve function. Study Design This is a randomized controlled animal study. Materials and Methods This experimental study was performed on 84 male Wistar albino rats. The rats were assigned to 12 groups each consisting of 7 animals. The groups were formed according to application of normal-dose DXM (group 1A-B), high-dose MP (group 2A-B), normal-dose MP (group 3A-B), MM (group 4A-B), and MM with high-dose MP combination therapies (group VA-B). Right sciatic nerve dissection was performed, and compound muscle action potential thresholds were recorded. The nerve was traumatized with the compression of a Jeweller forceps for 20 seconds. Posttraumatic thresholds were also recorded. The compound muscle action potential thresholds were recorded in the first and fourth weeks for the assigned groups. Then, the nerve was transected and prepared for electron microscopic and histopathologic examinations. Nitric oxide and malondialdehyde assessments were performed on both tissue and blood samples. Results Only the MM and MP+MM groups had satisfactory electron microscopic findings and were about to reach the tissue characteristics of the control animals. Despite the electrophysiologic recovery, the DXM group was found to have poor electron microscopic scoring. Conclusions Mycophenolate mofetil has been found to be beneficial in the treatment of traumatic nerve paralysis. Although a complementary investigation is needed, this immunosuppressive agent may be an alternative to corticosteroids for the selected cases where steroid therapy is contraindicated.