Genetic and Bioengineering

Purpose: To investigate the total polyphenolic and flavonoid contents, antioxidant power and cytotoxic activity of ethanol extracts of Turkish propolis (EEP).
Methods: The total polyphenolic and flavonoid contents of EEP were determined by spectrometric methods. Antioxidant power and cytotoxic activity of EEP were evaluated using ferric reducing antioxidant power (FRAP) and MTT assays, respectively.
Results: The total polyphenolic and flavonoid contents, and FRAP value of EEP were 124.6 ± 1.5 mg gallic acid/g sample dry weight , 42.0 ± 0.8 mg quercetin/g sample dry weight and 311.0 ± 2.5 mg trolox/g sample dry weight, respectively. EEP exhibited powerful cytotoxic effects against the five human cancer cell lines. The highest cytotoxic activity of Turkish EEP was demonstrated on PC-3 cell line (IC50 = 20.7±3.4 μg/mL).
Conclusion: The results demonstrate that EEP is a good source of antioxidant and a natural antitumor agent capable of reducing cancer cell proliferation.

Objective: Nut consumption has beneficial effects on protection for development of atherosclerotic process. Methods: Single intervention study design was used to determine the effects of hazelnut-enriched diet (1 g/kg/day) during 4 weeks period on atherogenic tendency of low-density lipoprotein (LDL) by evaluating susceptibility of LDL to oxidation, α-tocopherol content of LDL, LDL subfractions, plasma oxidized (ox) LDL, lipid and lipoprotein levels in normolipidemic healthy subjects (n=21). Statistical analysis was performed using paired t test, ANOVA for repeated measurements test, Pearson's and Spearman correlation analyses. Results: Lag time for oxidation (baseline 54.6±12.3 min, 15 th day 59.3±13.4 min, 30 th day 65.2±17.8 min, p=0.001) and α-tocopherol content of LDL (baseline 4.82±1.2 μg/mg LDL protein, 15 th day 4.88±1.4 μg/mg LDL protein, 30 th day 5.35±1.7 μg/mg LDL protein, p=0.02) were found to be increased while ox-LDL levels (baseline 57.2±16.2 U/L, 15 th day 51.2±13.6 U/L, 30 th day 48.2±14.2 U/L, p=0.001) decreased during the study period. Total cholesterol, LDL-cholesterol, apolipoprotein (apo) B and apo B/apo AI ratio were found to be significantly lower while apo AI was higher (p<0.05). In respect to LDL subfraction, ratio of large/small LDL was significantly increased at the end of the study (baseline 3.79±1.35, 15 th day 3.41±1.60, 30 th day 4.28±2.44, p= 0.046). Conclusion: Hazelnut-enriched diet may play important role in decrease in atherogenic tendency of LDL by lowering the susceptibility of LDL to oxidation and plasma ox-LDL levels, and increasing the ratio of large/small LDL beyond its beneficial effect on lipid and lipoprotein levels. (Anadolu Kardiyol Derg 2010; 10: 28-35)

Intensive use in daily life of devices with an electromagnetic field (EMF), such as radios, televisions, computers and mobile phones, results in constant exposure. The fact that more than 80% of people in many countries use mobile phones has further increased concerns about the effect of EMFs. The aim of this study was to investigate the effect of exposure to a 900-megahertz (MHz) EMF in the prenatal term on the newborn rat testicles. Six pregnant, female Sprague Dawley rats were equally divided into control (CG) and EMF (EMFG) groups. No procedure was performed on CG. EMFG was exposed to a 900 MHz EMF for 1 h during days 13-21 of pregnancy at the same hour every day. No procedure was performed on newborn female rats after birth, and the study continued with newborn male rats. Newborn CG (NCG, n: 10) male rats were obtained from mothers in the CG, and newborn EMFG (NEMFG, n: 10) male rats from mothers in the EMFG. All animals were sacrificed and the testicles extracted at postnatal day 21. Testicular tissue samples were evaluated histopathologically under light microscope. The TUNEL technique was used to determine apoptosis in testicles. Lipid peroxidation and DNA oxidation levels were evaluated biochemically. Histopathologically, NEMFG male rats exhibited irregularities in the seminiferous tubule basal membrane and epithelium of seminiferous tubules, immature germ cells in the lumen, and a decreased diameter of seminiferous tubules and thickness of epithelium. Apoptotic index, lipid peroxidation and DNA oxidation were higher in NEMFG male rats than in NCG. Our results showed that the newborn rat testicles exposed to 900 MHz EMF in the prenatal period has been adversely affected.

Background/aim: Propolis is a bee product with antioxidative, antimutagenic, and other beneficial properties, and it is used as a natural drug. It is rich in polyphenolic compounds. Its composition varies depending on the particular geographical region. Oxidative stress is caused by an imbalanced free radical production and antioxidant system. The effects of flavonoids on the expression of DNA repair enzymes have been examined previously; however, no study has investigated the effects of propolis. This study investigated the effects of ethanolic extracts of Turkish propolis (EEP) on the expression of DNA repair enzymes.

Genotypes have to be controlled periodically and necessary precautions should be taken in order to ensure that animal biodiversity is not lost. In this study, the genotypes of the Akkaraman, Karayaka and Morkaraman species from among Turkey's local sheep were examined at 7 different microsatellite loci. The average allele counts are close to each other and were determined as 8,905±0,483. Whereas the PIC values ranged from between 0,507-0,794 at an average of 0,705. The He and Ho values used to calculate genetic diversity were determined on average as 0,734±0,023 and 0,509±0,038 respectively. According to the Fisher-Exact test result, the populations are not in HWE (P<0.001). Whereas the F IS values that show interpopulation genetic diversity were determined as 0,353±0,092 (KRY), 0,328±0,095 (MRK) and 0,237±0,059 (AKK) respectively. In addition, Population Assignment and Principal Coordinate Analysis graphs were formed. When the pairwise F ST and N m , NeiuD, S H UA and G Statistics values used to calculate the genetic similarities and differences are examined, it is observed that the closest relations in terms of genetics is among AKK-KRY followed respectively by AKK-MRK and KRY-MRK.

Study objective: The primary aim of this study was to investigate whether IMA levels are helpful in the diagnosis of pulmonary embolism (PE). The secondary aim was to determine whether IMA was more effective alone or in combination with clinical probability scores in the diagnosis of PE. Thirdly, the sensitivity and specificity of IMA is compared with D-dimer both with and without clinical probability scores in patients with suspected PE. Methods: Consecutive patients presenting to the emergency department with suspected PE were prospectively recruited, and healthy volunteers were also enrolled as controls. D-dimer and IMA levels were measured for the entire study group. Wells and Geneva scores were calculated and s-CTPA was performed on all suspected PE patients. Results: The study population consisted of 130 patients with suspected PE and 59 healthy controls. Mean IMA levels were 0.362 ± 0.11 ABSU for Group A, the PE group (n = 75); 0.265 ± 0.07 ABSU for Group B, the non-PE group (n = 55); and 0.175 ± 0.05 ABSU for Group C, the healthy control group (p < 0.0001). At a cutoff point of 0.25 ABSU, IMA was 93% sensitive and 75% specific in the diagnosis of PE. PPV was 79.4% and NPV was 78.6%. Mean D-dimer levels were 12.48 ± 10.88 μg/ml for Group A; 5.36 ± 7.80 μg/ ml for Group B and 0.36 ± 0.16 μg/ml for Group C (p < 0.0001). The D-dimer cutoff point was 0.81 μg/ ml with a sensitivity of 98.9% and a specificity of 62.7%, PPV of 69.4% and NPV of 83.3%. The use of IMA in combination with Wells and Geneva clinical probability scores was determined to have a positive impact on these scores' sensitivity and negative predictive values. Conclusion: IMA is a good alternative to D-dimer in PE diagnosis in terms of both cost and efficiency. Used in combination with clinical probability scores, it has a similar positive effect on NPV and sensitivity to that of D-dimer. The PPV of IMA is better than D-dimer, but it is still unable to confirm a diagnosis of PE without additional investigation.

Background: Pulmonary embolism (PE) is a common condition, but the diagnostic strategy for the evaluation of suspected PE is somewhat controversial. Despite the use of various biochemical markers (such as D-dimer and C-reactive protein) and various probability calculation algorithms based on clinical findings for that purpose, there is still a need for more specific and practical markers in PE diagnosis. The aim of this study was to investigate the diagnostic value of ischemia-modified albumin (IMA) levels in the diagnosis of PE. Methods: This case-control study was performed in the emergency department between March and September 2006. The serum IMA levels of a total of 60 individuals, consisting of 30 PE patients who had been definitively diagnosed via spiral computed tomographic angiography and 30 healthy volunteers, were examined. Results: The measurement of IMA levels in patient plasma yielded mean values of 0.724 F 0.122 absorbance unit (ABSU) in the PE group and 0.360 F 0.090 ABSU in the control group. When plasma IMA levels in the PE group were compared with those in the control group, statistically significant increases in IMA were observed in the former (t = 13.19, df = 56, P b .0005). The value of 0.540 ABSU was calculated as the upper limit of reference interval. In the PE group, 97.7% (n = 29) had values exceeding 0.540 ABSU; none of the control subjects had values exceeding this cutoff value. Conclusions: In conclusion, our data suggest that IMA levels may be useful as a discriminative marker to exclude pulmonary embolism.

Background: Previous studies have demonstrated that ischemia-modified albumin (IMA) is a useful marker for the diagnosis of ischemic events. It was also recently demonstrated that IMA levels increase in the acute phase of cerebrovascular diseases. Yet the data regarding IMA levels in various types of cerebrovascular events are insufficient. The aim of this study was to evaluate IMA levels in various types of cerebrovascular events such as ischemic stroke, subarachnoid hemorrhage (SAH), and intracranial hemorrhage. Methods: This case-controlled study consisted of 106 consecutive patients, 43 with brain infarction (BI), 11 with brain hemorrhage (ICH), 52 with SAH, and a 43-member control group. We investigated whether there was a statistical correlation between these 3 groups and the control group. The relations among the 3 groups were also examined. Comparisons among groups were done with analysis of variance. Results: Mean serum IMA levels were 0.280 ± 0.045 absorbance units (ABSU) for BI patients, 0.259 ± 0.053 ABSU for ICH patients, 0.243 ± 0.061 ABSU for SAH patients, and 0.172 ± 0.045 ABSU for the control group.There was a statistically significant difference between the mean IMA levels of BI, ICH, and SAH patients and the mean control patient IMA levels (P b .0001). Conclusions: Ischemia-modified albumin levels are high in cerebrovascular diseases. Ischemiamodified albumin measurement can also be used to distinguish SAH from BI during the acute phase of cerebrovascular event in the emergency department.

Ischemia-modified albumin (IMA) is a sensitive marker of myocardial ischemia, skeletal muscle ischemia, pulmonary embolism, and stroke. However, there are no studies showing whether IMA increases in mesenteric ischemia. The aim of this study was to determine whether IMA was elevated in acute mesenteric ischemia. This case-controlled study was performed in an emergency department of a university hospital. The measurement of IMA levels in patient plasma yielded means of 0.264 ± 0.057 absorbance units (ABSU) in the thromboembolic occlusion of the superior mesenteric artery (SMA) group and 0.163 ± 0.025 ABSU in the control group. When plasma IMA levels in the thromboembolic occlusion SMA group were compared with those in the control group, statistically significant increases in IMA were observed in the occlusion group (P = .003). Findings indicating that IMA may have a place in the diagnosis of acute mesenteric embolism were obtained in this preliminary study. Further prospective studies are needed to see if IMA is clinically useful in the early detection of thromboembolic occlusion of the SMA.

Laurocerasus officinalis Roem. (Prunus laurocerasus l.) belongs to the genus Prunus and the family Rosaceae. It is a popular dark blue or black fruit growing particularly in the Black Sea region of Turkey. The plant is also known as cherry laurel in the English-language literature, and is grown in some Balkan, Western Europe, Middle Eastern and Mediterranean countries. This fruit is very popular in Turkey as a foodstuff and is consumed not only in fresh form, but also in the forms of jam, marmalade or fruit juice (1-4). It is regarded as a good source of nutrients due to the content of monosaccharides, ascorbic acid, dietary fiber, various minerals, and phenolics, such as chlorogenic, benzoic, vanillic, and caffeic acid (5). This fruit is used to treat several diseases in Turkish traditional medicine, such as eczema, sore throat, asthma, coughs, stomach ache, and hemorrhoids (1-4, 6). Recent studies have reported anti-inflammatory, antinociceptive, antioxidant, anti-atherosclerotic, and antidiabetic activities of L. officinalis, and these beneficial activities have been attributed to the compounds listed above (1, 4-7).

Objective: Cancer is one of the most deadly types of disease and evasion from apoptosis and unstoppable cell proliferation are accepted as its distinctive features. Many studies have evaluated the cytotoxic effect of different Morus species but, there is no study about cytotoxic effect of Morus rubra. In this study we aimed to evaluate phenolic composition, antioxidant properties and cytotoxic effect of acidified dimethyl sulfoxide extract of M. rubra (AMRE). Method: Antioxidant properties, phenolic composition and cytotoxic effect of AMRE were determined using spectrophotometric methods, HPLC, and MTT assay, respectively. Then, mechanisms of cytotoxic effect of AMRE on human prostate (PC-3) and lung (A549) cancer cells were examined in regard to cell cycle, apoptosis and mitochondrial membrane potential using flow cytometric methods. Results: Total phenolic content and reducing power values were 11.9 mg gallic acid equivalents and 42.9 mg trolox equivalents per g sample, respectively. Ascorbic and gallic acid were detected in AMRE as major antioxidant compounds. We determined that AMRE increased cell cycle arrest at G 1 phase and exhibited apoptotic features via decreasing mitochondrial membrane potential in both prostate and lung cancer cells. Conclusion: These findings demonstrate that M. rubra extract can affect the behavior of human prostate and lung cancer cells in vitro conditions, and this effect now needs to be investigated in vivo.

Cancer is the second leading cause of death and gastric cancer is the fourth most common cancer type worldwide. investigation of autoantibodies in cancer patients has been a popular research area in recent years. the aim of the current study was to investigate carbonic anhydrase i and ii (CA i and ii) autoantibodies in the plasma of subjects with gastric cancer based on the information and considerations of autoimmune relation of gastric cancer. Anti-CA i and ii antibody levels were investigated by elisA in plasma samples of fifty two patients with gastric cancer and thirty five healthy peers. Anti-CA i and ii antibody titers of the gastric cancer group were significantly higher compared with the control group (p = 0.004, p = 0.0001, respectively). plasma anti-CA i levels of the metastatic group were lower than the non-metastatic group and this difference was found statistically significant (p < 0.05), but there was no statistical difference between plasma anti-CA ii levels of the groups. CA i and ii autoantibody titers in patients with gastric cancer were found higher compared to healthy subjects and the results suggest that these autoantibodies may be involved in the pathogenesis of gastric cancer.

Mn(II), Co(II), Ni(II), Cu(II), and Zn(II) transition metal complexes of 2-hydroxy-5-[(E)-(4-phenyl) diazenyl] benzaldehyde oxime and 2-hydroxy-5-[(E)-(4-nitrophenyl) diazenyl] benzaldehyde oxime ligands were synthesized and characterized through NMR, IR, ESI mass, and UV analysis. DNA binding abilities of the complexes were revealed using a UV-Vis spectrophotometer with the absorption titration and competitive binding techniques. Hydrolytic and oxidative DNA cleavage activities under different conditions were also proved. Topoisomerase I inhibition efficiencies and in vitro free radical scavenging activities of all complexes were examined. Finally, the selective cytotoxic potentials of all complexes were evaluated in human colon cancer, normal colon, and fibroblast cell lines using the water-soluble tetrazolium salt (WST-1) assay. The complexes had the ability to intercalate into stacked base pairs of DNA and topoisomerase I activity was reasonably inhibited in their presence in 0.4 mM concentrations. The abilities for scavenging of DPPH and hydroxyl radicals were found to be higher than those of known standard antioxidants (ascorbic acid, butylated hydroxyanisole, and mannitol). The results obtained from the cytotoxicity experiments are especially promising for further research, which must be carried out for the evaluation of the studied complexes as anticancer drugs.

Breast cancer is the second most common cancer type among women worldwide. Chemotherapy is the most commonly used strategy against in treatment of breast cancer, but chemotherapeutics also have toxic effects on healthy cells. Natural products have low toxicity, so numbers of chemotherapeutic agents extracted from natural products for treatment of various types of cancers have been increasing for yearly. Propolis is a bee product and contains various polyphenolic compounds. It has been shown that polyphenols affect various signal pathways, such as inhibit cell proliferation, and activate apoptosis, mitochondrial dysfunction. In this study, we aimed to evaluate the effect of Turkish propolis ethanolic extract (EEP) on mitochondrial membrane potential (MMP) alterations, the levels of p53-Ser46 and p53-Ser15 protein in human breast cancer (MCF-7) cells. MMP alterations, protein levels of p53-Ser46 and p53-Ser15 were determined using spectrofluorometric analysis and western blot method, respectively. Results: EEP significantly reduced MMP in dose dependent manner in MCF-7 cells. Moreover, EEP caused increase p53-Ser46 and p53-Ser15 protein levels at 72 h in these cells. Mitochondrial dysfunction is as an important constituent of apoptosis process. EEP may have apoptotic effects in MCF-7 cells through loss of MMP depend on increased phosphorylation of p53.