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In this study, firstly, five novel vitamin K3 analogues were synthesized by the reactions of vitamin K3 (2-methyl-1,4-naphthoquinone, also known as menadione) with some aromatic and heterocyclic ring substituted nucleophiles such as 2,4-dimethoxyaniline, 4-methoxyaniline, 4-benzylpiperidine and 1-(2-aminoethyl)piperazine in ethanol/Na2CO3, and 1-(diphenylmethyl)piperazine in chloroform/triethylamine (TEA) at room temperature. Their structures were elucidated by Fourier transform infrared spectroscopy (FT-IR), 1H nuclear magnetic resonance (1H NMR), attached proton test nuclear magnetic resonance (APT-NMR) and mass spectrometry (MS). Secondly, in vitro cytotoxic effects of vitamin K3 analogues were investigated by MTT assay against three cancer cell lines (HeLa, U87 MG, and MCF-7) to evaluate their anticancer activity and a human embryonic kidney cell line (HEK-293) to check their cancer cell selectivity. One of the compounds, namely 2-((2,4-dimethoxyphenyl)amino)-3-methylnaphthalene-1,4-dione, was found to inhibit the growth of HeLa cervical cancer cells selectively, even better than vitamin K3, at a non-toxic concentration for healthy cells. The selectivity index of this compound for HeLa cells was calculated approximately as 3. Vitamin K3 was more effective against U87 MG and MCF-7 cells than its derivatives, moreover it was the only compound, which was significantly toxic to breast cancer cells, but its selectivity was poor. Furthermore, anticancer properties of piperazine derivatives of vitamin K3 were investigated by us for the first time in this study.

The prevalence of allergic asthma is increasing on a global scale, reflecting changes in air pollution, climatic changes, and other environmental stimulants. In allergic conditions, oxidative stress occurs as a result of immune system activation. Oxidation of cholesterol leads to the formation of oxysterols. The main purpose of the study was to compare plasma levels of two oxysterols, namely 7-ketocholesterol (7-KC) and cholestane-3β, 5α, 6β-triol (C-triol), and a lipid peroxidation product, malondialdehyde (MDA) in allergic asthma patients with those of healthy controls, in order to provide information about the involvement of lipid peroxidation in allergic asthma. Oxysterols were quantified by LC-MS/MS in plasma samples of 120 asthma patients (90 females + 30 males) and 120 healthy controls (matched by age and sex). Plasma MDA level was analyzed by a spectrophotometric method. Plasma 7-KC (39.45 ± 20.37 ng/mL) and C-triol (25.61 ± 10.13 ng/mL) levels in patients were significantly higher than in healthy subjects (17.84 ± 4.26 ng/mL and 10.00 ± 3.90 ng/mL, respectively) (P < 0.001). Plasma MDA levels were also higher in asthmatic patients (4.98 ± 1.77 nmol/mL) than in healthy controls (1.14 ± 0.31 nmol/mL) (P < 0.001). All data support that lipid peroxidation products are involved in allergic asthma. Oxysterols were quantified for the first time in allergic asthma. Since the high plasma 7-KC and C-triol levels of allergic asthma patients correlate with high IgE levels, detection of these oxysterols by LC-MS/MS may be helpful in the clinical monitoring of allergic asthma. Current data may also lead to new approaches for the prevention, diagnosis, and treatment of the disease. Supplemental data for this article is available online at at.

The Jurinea Cass. is one of the most important genera within Asteraceae and it comprises about 250 species in total. This genus is known for its numerous biological activities such as antioxidant, antimi-crobial, antilipid peroxidation, anticholinesterase, antileishmanial activities. The aim of this study was to determine chemical composition and biological activities of ethanol and n-hexane extracts of three differ-ent Jurinea species. For this purpose, different parts of J. mollis, J. cadmea and J. pontica were extracted and totally six n-hexane and six ethanol extracts were obtained. Fatty acid content of n-hexane extracts was determined by GC-FID whereas phenolic and flavonoid content of ethanol extracts by LC-HRMS. Palmitic acid (16:0) was detected as the most abundant fatty acid in all n-hexane extracts with the rates rang -ing from 42.16%-55.08%, except flowers of J. mollis (JMF) and J. cadmea (JCF). LC-HRMS analysis showed the rutin content of all extracts was higher than other flavonoids, except of J. cadmea flowers, whereas apigenin-7-glucoside was found the most abundant in JCF. Cytotoxic effects of the extracts on HeLa and HEK-293 cells were determined by MTT method, and antioxidant activities were evaluated by DPPH and CUPRAC assays. Ethanol extract ofJ. mollis flowers significantly inhibited cancerous HeLa cells, with the IC50 value of 9.683 mu g/mL while it was more less toxic on healthy HEK-293 cells. Ethanol extracts of J. mollis flowers and J. mollis steams-leaves (JMSL) showed the highest antioxidant activity by a DPPH inhibition % of 45.516 +/- 2.497 and 56.671 +/- 1.496, respectively. JMF and JMSL have also the highest CUPRAC values (0.880 +/- 0.067 and 1.085 +/- 0.152 mmol TR/g DWE, respectively). Total flavonoid content was determined using aluminum chloride colorimetric assay while total tannin and phenolic content by Folin Chiocalteu's reagent. Results showed that JMSL has the highest total phenolic (108.359 +/- 6.241 mg GAE/ G DWE) and flavonoid (32.080 +/- 4.385 mg QE/ g DWE) contents whereas JMF has the highest tannin content (121.333 +/- 17.889 mg TAE/ g DWE). In the light of these results, various parts of Jurinea species may be regarded as alternative sources for cytotoxic and/or antioxidant flavonoids, phenolics and unsaturated fatty acids that can arouse the interest of pharmaceutical, food and cosmetic industries. (C) 2021 Elsevier B.V. All rights reserved., National Natural Science Fund for Distinguished Young Scholars [30325044]; Key Grant of Chinese Ministry of Education [306010]; ChinaOcean Resource RD Association [DYXM-115-02-2-13]; 973 Program [2010CB833802]; Research Council of Turkey, This work was partially supported by the National Natural Science Fund for Distinguished Young Scholars to Y.S. (30325044), the Key Grant of Chinese Ministry of Education (306010), ChinaOcean Resource R&D Association (Grant DYXM-115-02-2-13) and 973 Program (2010CB833802). The authors also would like to thank Ege University, Research and Application Center of Drug Development and Pharmocokinetics for GC-FID analysis. G. Sumer Okkali isthankful to Research Council of Turkey for scholarship.

IntroductionMorus alba (white mulberry) pollen is an aero-allergen source that can trigger allergic diseases. Cobalamin-independent methionine synthase (MetE) in M. alba pollen has been proved to be one of the major allergens for some patients living in Istanbul (Turkey). The aim of the present study was the recombinant production and identification of MetE (Mor a 2), a novel allergen from M. alba pollen. The IgE binding reactivity of rMor a 2 produced for the first time was evaluated and some structural features were investigated by in silico methods to better understand its immunogenicity.Material and methodsThe gene encoding Mor a 2 was cloned in fission yeast, Schizosaccharomyces pombe ura4-D18h- strain, using pSLF1073 vector. This is the first report of the production of recombinant pollen allergen in S. pombe. After the purification, immunoreactivity of rMor a 2 was confirmed by immunoblotting using sera of patient allergic to M. alba pollen. Besides, B-cell epitopes of rMor a 2 were predicted using various bioinformatic tools, namely Bioinformatics Predicted Antigenic Peptides, BepiPred 2.0 and Immune Epitope Database whereas T-cell epitopes were estimated using NetMHCIIpan-3.2 and NetMHCII 2.3 servers.ResultsThe immunoblotting analysis yielded 11 of 11 positive reactions to rMor a 2. In silico predictions exerted seven B-cell epitopes (22-33, 384-394, 407-423, 547-553, 571-577, 671-678, 736-741) and seven T-cell epitopes (54-62, 161-170, 197-205, 347-358, 622-630, 657-665, 756-764).ConclusionsThese findings may help the use of rMor a 2 in the diagnosis and treatment of allergic diseases associated with M. alba and/or MetE.

There is evidence that reactive oxygen species, especially free radicals, produced during the immune and inflammatory response may play important roles in the development of asthma.We aimed to evaluate the levels of certain oxidative stress biomarkers and antioxidant capacity in asthma patients with different asthma control levels in comparison to healthy subjects.A total of 120 adult allergic asthma patients and 120 healthy individuals were included in this study. Using spectrophotometric methods, we analyzed two oxidative stress markers, levels of malondialdehyde (MDA) and protein carbonyls (PC), as well as reduced glutathione (GSH), total antioxidant capacity (FRAP) and catalase activity as critical antioxidant defense parameters in the blood samples of allergic asthma patients and healthy controls. The patients were divided into 3 subgroups according to asthma control test (ACT) results: totally controlled (TCG), partially controlled (PCG) and uncontrolled (UCG) subgroups. All biomarkers were compared between the three patient subgroups, as well as between total asthma patients and control subjects.There were remarkable differences between the control group and the combined patient group for all parameters. A significant increase in MDA and PC, especially in the UCG (This research demonstrates the presence of severe oxidative stress, considering the increase in lipid peroxidation and protein oxidation, in patients with allergic asthma, even under controlled conditions.

Methotrexate (MTX), an analog of folic acid (FA), is a drug widely used in cancer treatment. To prevent its potential toxicity and enhance therapeutic efficacy, targeted drug delivery systems, especially nanotechnology-folate platforms, are a central strategy. Gold nanoparticles (AuNPs) are promising candidates to be used as drug delivery systems because of their small particle sizes and their inertness for the body. In this study, glutathione (GSH)-coated FA-modified spherical AuNPs (5.6 nm) were successfully synthesized, and the anticancer activity of novel MTX-loaded (MTX/Au-GSH-FA) NPs (11 nm) was examined. Dynamic light scattering (DLS) and transmission electron microscopy (TEM) results showed that MTX/AuNPs possess spherical morphology, nanoscaled particle size, narrow size distribution, and good stability. In vitro studies showed that cytotoxicity of MTX/Au-GSH-FA to folate receptor-positive (FR+) human brain (U-87 MG) and cervical (HeLa) cancer cells enhanced significantly (∼3 and ∼10 fold, respectively) compared to free MTX while there was no significant effect in FR-negative human cell lines A549 (lung carcinoma), PC3 (prostate carcinoma), HEK-293 (healthy embryonic kidney). Moreover, the receptor specificity of the conjugate was shown by fluorescent microscopic imaging. In conclusion, these results indicate that the synthesized novel MTX/Au-GSH-FA NP complex seems to be a good candidate for effective and targeted delivery in FR+ cancer therapy.

Gliomas from glial cells of the brain are highly aggressive neoplasms. Traditional treatment has very limited effectiveness, and the prognosis is still poor. One of the major obstacles to therapy of gliomas is the resistance to treatment and the increased heat shock protein (Hsp) levels have a great effect on this. Overexpression of many Hsp, especially Hsp27, Hsp70 and Hsp90, is observed in gliomas. There is a relationship between Hsp expression levels and the tumor grade, and their functions aides the advancement of cancer. Therefore, it has become important to target Hsp in glioma treatment. Today, the disciplines of gene therapy (GT) have been experiencing a revolutionary growth in the field of cancer treatment. Developments in the field of genome-editing have made the GT methods a powerful potential tool for downregulation of Hsp and the breakdown of gliomas’ treatment resistance. In this respect, the GT appears to be a promising innovative approach against gliomas. In this chapter, we reviewed the role of overexpressed Hsp in gliomas and the importance of the GT methods for silencing of them. We emphasized the principles, expectations and limitations of the methods, and highlighted the potential of the GT methods for Hsp targeted treatment of gliomas.

Mertoglu, Elif/0000-0002-3606-4722; ARDA, NAZLI/0000-0002-1043-5652; Aydinli, Serdar Goksin/0000-0002-2681-6011; UCAR, Evren ONAY/0000-0003-1152-4881; Sayil, Makbule Cigdem/0000-0001-9611-3533; Kacmaz, Aysecik/0000-0001-8420-6855 WOS:000471119600003 In the course of biological properties of quinone derivatives, the N(H)-, S- and S,S-substituted-1,4-naphthoquinones were synthesized by reactions of 2,3-dichloro-1,4-naphthoquinone with different amines (2-morpholinoaniline, tert-butyl 4-aminobenzoate, 4-tert- butyl-benzylamine, N-(3-aminopropyl)-2-pipecoline, 2-amino-5,6-dimethylbenzothiazole, N,N'-diphenyl- p-phenylenediamine) and thiolat (sodium 2-methyl-2-propanethiolate). All new products were characterized by MS-ESL UV-Vis, FT-IR, H-1 NMR, C-13 NMR. The antiproliferative activities of these compounds on human cervical cancer (HeLa) cells were evaluated by MTT (3[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay. Although all derivatives inhibited cell growth, the most active compound was 2-(tert-butylthio)-3-chloronaphthalene-1,4-dione 5 (IC50 =10.16 mu M) against the HeLa cells. Scientific Research Projects Coordination Unit of Istanbul University-Cerrahpasa [36017, YADOP-32026] The authors would like to express their gratitude to Scientific Research Projects Coordination Unit of Istanbul University-Cerrahpasa for financial support (Project Numbers: 36017, YADOP-32026).

Ucar, Evren Onay/0000-0003-1152-4881; Akdeniz, Nilgun/0000-0002-6208-3193; ARDA, NAZLI/0000-0002-1043-5652; KARLIOGLU KILIC, Nurgul/0000-0002-6255-6819; Bourgoin-Voillard, Sandrine/0000-0001-6511-0612 WOS:000506666400003 PubMed ID: 30447650 Background: Tree pollens are well-known aeroallergens all over the world. Little is known about the allergenicity of Morus alba (white mulberry) pollen. Objective: We aimed to explore the potential allergens of this pollen and its clinical relevance in tree pollen allergic patients living in Istanbul, Turkey. Methods: Twenty three seasonal allergic rhinitis patients with a confirmed tree pollen allergy and 5 healthy control subjects underwent skin prick and nasal provocation tests with M.alba pollen extract. The pollen extract was then resolved by gel electrophoresis, and immunoblotted with sera from patients/control individuals to detect the potential allergenic proteins. The prevalent IgE binding proteins from ID-gel were analyzed by MALDI-TOF/TOE Results: Eleven out of 23 patients were reactive to the extract with skin prick tests. Seven of those patients also reacted positively to the nasal provocation tests. The most common IgE-binding pollen proteins were detected between 55-100 kDa, and also at molecular weights lower than 30 kDa for some patients. Mass spectrometry analyses revealed that the principal IgE-binding protein was methionine synthase (5-methyltetrahydropteroyltriglutamate homocysteine methyltransferase), which is then proposed as a novel allergen in M.alba pollen. Conclusion: This study provides the first detailed information for the potential allergens of Morus alba pollen of Istanbul. Methionine synthase with an apparent molecular weight of 80 to 85 kDa has been recognized as one of the allergens in Morus alba pollen for the first time. Research Fund of Istanbul UniversityIstanbul University [3063, 30217]; Bio-Health Computing Erasmus Mundus program [512383-1-2010-1-FR] This study was supported by the Research Fund of Istanbul University (Project no. 3063 and Project no. 30217) and the Bio-Health Computing Erasmus Mundus program (512383-1-2010-1-FR).

Tree pollens are well-known aeroallergens all over the world. Little is known about the allergenicity of Morus alba (white mulberry) pollen.We aimed to explore the potential allergens of this pollen and its clinical relevance in tree pollen allergic patients living in Istanbul, Turkey.Twenty three seasonal allergic rhinitis patients with a confirmed tree pollen allergy and 5 healthy control subjects underwent skin prick and nasal provocation tests with M.alba pollen extract. The pollen extract was then resolved by gel electrophoresis, and immunoblotted with sera from patients/control individuals to detect the potential allergenic proteins. The prevalent IgE binding proteins from 1D-gel were analyzed by MALDI-TOF/TOF.Eleven out of 23 patients were reactive to the extract with skin prick tests. Seven of those patients also reacted positively to the nasal provocation tests. The most common IgE-binding pollen proteins were detected between 55-100 kDa, and also at molecular weights lower than 30 kDa for some patients. Mass spectrometry analyses revealed that the principal IgE-binding protein was methionine synthase (5-methyltetrahydropteroyltriglutamate homocysteine methyltransferase), which is then proposed as a novel allergen in M.alba pollen.This study provides the first detailed information for the potential allergens of Morus alba pollen of Istanbul. Methionine synthase with an apparent molecular weight of 80 to 85 kDa has been recognized as one of the allergens in Morus alba pollen for the first time.

Heat shock response is one of several survival pathways that protects cells against harsh conditions. This response mechanism, which is evolutionarily protected in all organisms, enhances the expression of heat shock proteins (HSP) that show protective properties for cells under stress conditions. High expression of many HSP is observed in cancer, and their functions aides the advancement of disease. It is known that overexpression of HSP70, a member of HSP family, in cancerous cells has been closely associated with tumor cell proliferation, apoptosis inhibition, enhanced migration and metastasis and drug resistance promotion. Therefore, targeting HSP70 in cancer treatment is very important. One of the best-studied inhibitors known for HSP70 is quercetin that is widely distributed flavonoid in the plant kingdom. Several in vivo and in vitro studies have reported the efficacy of quercetin in reducing elevated HSP70 levels in cancer therapy. It has become a focal point as an anticancer agent because of the induction of apoptosis in many different cancer cells. In this chapter, we reviewed the role of HSP70 in different cancer types and the suppressive effect of quercetin on expression of HSP70 family members. Moreover, we emphasized molecular mechanisms targeted by quercetin in cancer and its relationship to Hsp70.

Beta cell mass regulation represents a critical issue for understanding and treatment of diabetes. The most important process in the development of diabetes is beta cell death, generally induced by glucotoxicity or glucolipotoxicity, and the regeneration mechanism of new beta cells that will replace dead beta cells is still not fully understood. The aim of this study was to investigate the generation mechanism of new beta cells by considering the compensation phase of type 2 diabetes mellitus. In this study, pancreatic islet derived mesenchymal stem cells (PI-MSCs) were isolated from adult rats and characterized. Then, beta cells isolated from rats were co-cultured with PI-MSCs and they were exposed to glucotoxicity, lipotoxicity and glucolipotoxicity conditions for 72 hr. As the results apoptotic and necrotic cell death were increased in both PI-MSCs and beta cells especially by the exposure of glucotoxic and glucolipotoxic conditions to the co-culture systems. Glucotoxicity induced-differentiated beta cells were functional due to their capability of insulin secretion in response to rising glucose concentrations. Moreover, beta cell proliferation was induced in the glucotoxicity-treated co-culture system whereas suppressed in lipotoxicity or glucolipotoxicity-treated co-culture systems. In addition, 11 novel proteins, that may release from dead beta cells and have the ability to stimulate PI-MSCs in the direction of differentiation, were determined in media of glucotoxicity or glucolipotoxicity-treated co-culture systems. In conclusion, these molecules were considered as important for understanding cellular mechanism of beta cell differentiation and diabetes. Thus, they may be potential targets for diagnosis and cellular or therapeutic treatment of diabetes.

Indomethacin is a member of the non-steroidal anti-inflammatory drug (NSAID) class, which has great potential for use in the treatment of glioma. However, it induces the generation of reactive oxygen species (ROS) and causes molecular damage while inducing its effects. Vitamin E is widely used in the complementary therapy of cancers. The main goal of the present study was to investigate the effects of α-tocopheryl succinate (α-TOS) against the oxidative damage induced by indomethacin in C6 glioma cells. Cells were treated with 10 μM α-TOS alone or in combination with 200 μM indomethacin for two days. The intracellular ROS level, molecular damage as revealed by lipid peroxidation and protein carbonyl formation, and the COX activity in C6 glioma cells were measured. Treatment of the cells with α-TOS and indomethacin, alone or in combination, caused the levels of ROS generation and protein damage to increase, but protected against lipid peroxidation and reduced COX activity.

WOS: 000349005800018 PubMed ID: 25266234 Oxysterols, oxygenated derivatives of cholesterol, are found abundantly in the plasma and atherosclerotic plaques, a common risk factor for thoracic aortic aneurysms (TAAs). Among the oxysterols, namely 7-ketocholesterol (7-KC) and 25-hydroxycholesterol (25-OHC), lead both to induction of reactive oxygen species (ROS) in cells and to apoptosis in smooth muscle cells (SMCs) probably due to increased oxidative stress. Since loss of SMCs through apoptosis is a major event in TAA formation, it is important to understand the molecular pathways of apoptosis in response to ROS in TAAs. Very little is known about the effect of oxysterols on TAA SMCs. Therefore, we investigated molecular pathways participating in the oxysterol induced cell death of TAAs. Our results showed that TAA SMCs died mainly as a result of apoptosis as suggested by cellular shrinkage, blebbing, DNA condensation/fragmentation in response to oxysterol treatment. There was no significant difference in oxysterol induced cell death between TAA and control SMCs. Addition of antioxidant molecules prevented cell death, hence ROS appears to be involved in the apoptosis of these cells. While oxysterol treatment increased caspase 3 activity, cell death was not rescued in its absence. Efficient silencing of other targets including apoptotic proteins (p53, Bax), and survival proteins (Akt1, Akt2) showed that apoptosis can occur through p53, and Bax independent pathways. Silencing Akt1 or Akt2 did not lead to further cell death. These results indicate that oxysterols can induce several cell death pathways in TAA SMCs. FP7 Project entitled "Fighting Aneurysmal Disease" (FAD) [200647] Istanbul University Department of Scientific Research Projects [10241] This work was supported by the FP7 Project entitled "Fighting Aneurysmal Disease" (FAD), Grant No: 200647 and Istanbul University Department of Scientific Research Projects, entitled "Investigation of Gene Expressions Related to Oxidative Stress in Human Thoracic Aort Aneurysms", Grant No: 10241.

Whey is used as an additive in food industry and a dietary supplement in nutrition. Here we report a comparative analysis of antioxidant potential of whey and its fractions. Fractions were obtained by size exclusion chromatography, before and after enzymatic digestion with pepsin or trypsin. Superoxide radical scavenging, lipid peroxidation inhibition and cupric ion reducing activities of different fractions were checked. Peptides were detected by SDS-PAGE and GC-MS was used to determine carbohydrate content of the fractions. All samples showed antioxidant activity and the second fraction of the trypsin hydrolysate showed the highest superoxide radical scavenging activity. CUPRAC value of this fraction was two-times higher than that of whey filtrate. The first fraction of the pepsin hydrolysate was the most effective inhibitor of lipid peroxidation. Each sample exhibited a different polypeptide profile. Different percentages of carbohydrates were identified in whey filtrate and in all second fractions, where galactose was the major component. (C) 2013 Elsevier Ltd. All rights reserved.

Initially discovered as a group of proteins showing significantly higher expression in response to heat stress, Heat shock proteins (HSPs) have gained considerable appreciation. Overwhelmingly increasing scientific evidence has highlighted the role of these proteins as molecular chaperones which trigger protein holding and folding thus facilitating freshly synthesized protein/s to achieve mature and biologically active conformation. It is becoming progressively more understandable that HSPs are involved in post-translational modification of proteins of signaling cascades, modulation of apoptosis related proteins, assembly and disassembly of transcriptional machinery. Recently emerging functional and structural data has provided new insights related to biochemical regulation of HSPs and the structural dynamics used by these proteins to act on a diverse client repertoire.

Objectives: Palladium complexes are potent and less toxic molecules in comparison to other metal based agents. Here, we characterized two palladium(II) saccharinate complexes with terpyridine for their cell cycle specificity. Materials and methods: Cells were arrested at Gl, Gl/S boundary or mitosis using mimosine, double-Thymidine block, aphidicolin, nocodazole or colcemid, and evaluated based on morphology and flow cytometry. Synchronized cells were treated with the Pd(II) complexes, and viability was measured via MTT assay. Results: While treatment of arrested cells with the Pd(II) complexes resulted in no significant change in cell death in HCT-116 and MDA-MB-231 cells, HeLa cells were more sensitive in S/G1. The main form of cell death was found to be apoptosis. Conclusions: Pd(II) complexes appear to be cell-cycle non-specific, while cell line dependent differences may be observed. Cells die through apoptosis regardless of the cell cycle stage, which makes these complexes more promising as anti-cancer agents. İstanbul Üniversitesi - 36696

Oxidative DNA damage is an inescapable consequence for cells constantly exposed to oxidative stress derived from normal metabolic processes and from environmental factors. Phenolic compounds, which have strong antioxidant activity, prevent DNA damage by protecting the cells against harmful effects of oxidative stress. In this study, the effect of virgin olive oil phenolic extract (OOPE) was investigated on H2O2-induced mitochondrial DNA (mtDNA) and nuclear DNA (nDNA) damage in HeLa cells. DNA damage was assessed in mitochondria and two nuclear regions by using quantitative PCR (QPCR) assay. The cells were pre-treated with non-cytotoxic doses of OOPE for 4 h, and DNA damage was determined. OOPE alone does not change the steady-state level of DNA damage. The oxidative stress generated with 750 μM H2O2 caused two times greater damages in mtDNA compared to nDNA, which included the nonexpressed β-globin region (1.507±0.110 lesions/10 kb) and the expressed APEX1 gene (1.623±0.243 lesions/10 kb) with respect to the control region. When cells were preincubated with OOPE for 4 h, nDNA damage under stress condition was completely inhibited; however, mtDNA damage was not affected by this procedure. These results suggest that OOPE has a protective effect against nDNA damage in HeLa cells.

Extracts of mistletoe (Viscum album) are intensively used in complementary medicine, but their mechanisms are not fully understood in most cases, and the effects on metabolism have not been investigated in detail. However, some biologically active natural products are well known to provoke unexpected cellular responses. They reduce overexpression of heat shock proteins (Hsps) in cancer cells. The aim of the current study was to determine whether methanolic extract of V. album, which possesses antioxidant activity, has an effect on expression levels of Hsp27 and 14-3-3 proteins in a C6 glioma cell line. For the first time, the apoptosis-inducing effect of this extract was also determined via caspase-3 activation in the cells. Overexpression of Hsps was induced by heat shock at 42°C for 1 h. Expression levels of Hsp27 and 14-3-3 proteins were determined using Western blot analysis. The apoptosis-inducing effect was also evaluated via caspase-3 activation in C6 glioma cells. Pretreatment of the cells with a nontoxic dose (100 μg/mL) of V. album extract before heat shock significantly reduced expression levels of Hsp27 (73%) and 14-3-3β (124%), 14-3-3γ (23%), and 14-3-3ζ (84%) proteins. Pretreatment with the extract before heat shock increased apoptosis via caspase-3 activation (60%) in C6 glioma cells. This result suggested that the methanolic extract of V. album downregulates expression of Hsp27 and 14-3-3 chaperone proteins and induces apoptosis, which warrants further exploration as a potential bioactive compound for cancer therapy.

Background: Many species of the genus Centaurea have been reported to be used in Anatolian folk medicine. In the present study, methanolic and aqueous extracts of three species of Centaurea L. (C. calcitrapa subsp. calcitrapa, C. ptosimopappa and C. spicata) have been assessed for antioxidant and cytotoxic activities. Materials and Methods: Antioxidant activity was investigated by DPPH and superoxide anion scavenging and anti-lipid peroxidation assays. Cytotoxic effect on HeLa and Vero cells were investigated by MTT assay. Results: Superoxide anion scavenging activities of aqueous extracts were higher than those of methanolic extracts, and the highest activity was detected in the aqueous extracts of C. calcitrapa subsp. calcitrapa with the IC 50 value of 25.91 μg/mL. The methanolic extract of C. ptosimopappa showed the strongest anti-lipid peroxidation activity. Total phenolic contents (mg gallic acid equivalent per gram) were ranged between 17.25 and 120.50 and the highest value was detected in aqueous extract of C. ptosimopappa. The highest cytotoxic activity was observed in the methanolic extract of C. calcitrapa subsp. calcitrapa on both cell lines with the IC 50 values of Conclusions: According to the results of activity tests, the selected Centaurea species, especially C. calcitrapa subsp. calcitrapa can be regarded as a potential source of natural antioxidants in the prevention and treatment of diseases in which oxidants or free radicals are implicated.

Cu,Zn superoxide dismutase (SOD1) is an antioxidant enzyme that catalyzes the removal of superoxide radicals generated in various biological oxidations. Amyotrophic lateral sclerosis (ALS) is one of the most common neurodegenerative disorders, occurring in families (FALS) and sporadically (SALS). FALS and SALS are distinguishable genetically but not clinically. More than 100 point mutations in the human SOD 1 gene have been identified that cause FALS. In order to determine the effects of mutant SOD protein, we first cloned wild-type and A4V mutant human SOD1 into Schizosaccharomyces pombe. This study shows viabilities and some antioxidant properties including SOD, catalase, proteasomal activity, and protein carbonyl levels of transformants in SOD1 deleted strain (MN415); and its parental strain (JY741) at different stress conditions. There was no more oxidative damage in the human mutant SOD carrying the transformant strain compared with other strains. These results may help to explain whether ALS progresses as a consequence of cellular oxidative damage.

The apolar constituents of four Dianthus (Caryophyllaceae) species were determined by GC-MS. Palmitic, linoleic, and oleic acids were detected as dominant components in all species. D. elegans d’Urv. var. elegans had the highest antioxidant activity. All four species also showed considerable antimicrobial activity against S. epidermidis and C. albicans.

Heterologous expression systems can be utilized to great advantage in the study of cytochrome P450 enzymes. P450 3A4 is one of the major forms of cytochrome P450 found in liver. It is also involved in the metabolism of numerous widely used drugs and xenobiotics. In the present study human liver cytochrome P450 3A4 gene was transferred into the fission yeast Schizosaccharomyces pombe via two different S. pombe expression vectors carrying thiamine repressible promoter — nmt1 (pREP42) and constitutive promoter — adh1 (pART1). Heterologously expressed cytochrome P450 3A4 was detected in the cells grown in minimal (EMM) or rich medium (YEL) containing 0.5% (w/v) glucose. A typical cytochrome P450 peak for 3A4 was observed at 448 nm in microsomal fraction. The presence of heterologous expression of 3A4 form was also determined by SDS-PAGE and it molecular mass was identified as 52 kDa. The enzyme activity was confirmed by HPLC analysis, using testosterone as substrate.

WOS: 000379860300018 Over the last decade, comparative molecular profiling studies between compatible and incompatible plant-pathogen interactions have shown that susceptible response of the host to a pathogen requires factors that promote disease development. In this study, we examined proteome profiles during a compatible interaction between wheat and stripe rust. A 2D-LC system (ProteomeLab PF2D) was used for protein separation and to compare the proteome from infected and control samples. More than 700 protein peaks at each time point were compared between pathogen- and mock-inoculated samples. Selected proteins, with significant differences in abundance were identified by nanoLC-ESI- MS/MS and generated spectra were searched against the wheat protein databases from UniProt, and NCBI and the Puccinia database from The Broad Institute. In total, the identified proteins comprised of 62 % wheat and 38 % Pst proteins. All identified proteins were searched by bioinformatics-based algorithms to detect their subcellular localization and signal peptide motifs which have the potential to catch the candidate effector proteins. The wheat proteins were classified based on their function. Although a compatible interaction, many wheat proteins, such as antioxidants, PRs and cold-responsive proteins, are implicated in defense and stress tolerance. On the pathogen side, 64 proteins were identified, and included some important pathogenicity proteins that can play role in pathogen virulence and suppress the host defense. In addition, we discovered that nine proteins have a signal sequence and three of the hypothetical fungal proteins, PGTG_11681T0, PGTG_07231T0 and CBH50687.1, have been tentatively identified as candidate effectors. TUBITAKTurkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [109 T293] This study was supported by TUBITAK, COST programme 109 T293 project. We thank Kadir Akan, Aysc Yildiz and Lutfi Cetin for their help during plant inoculation and sampling. We also thank Abdulmecit Gokce and Yavuz Ozturk for their technical support for PF2D as well as Konca Bulut and Rahmi Buyukkeskin for their experimental assistance.

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Zinc is a metal which is a cofactor in many enzymes and a structural element in zinc finger motifs those are important in relation between DNA and regulator proteins. Little is known about uptake, distribution, toxicity and detoxification of zinc ions in cells. In this study, zinc toxicity and detoxification levels have been compared in wild type and Cu/Zn superoxide dismutase mutant (sod1Δ) cells of the fission yeast Schizosaccharomyces pombe. We evaluated the toxic levels of zinc, total zinc content, lipid peroxidation levels and catalase activities for both strains which were grown in medium containing different concentrations of zinc. sod1Δ mutant showed important growth retardation and has higher lipid peroxidation and catalase activities than wild type. Cu/Zn superoxide dismutase (SOD1) activity of wild type cells was markedly increased when they were treated with elevated levels of zinc. SOD1 mRNA level also significantly increased when the cells treated with higher concentrations of zinc. These results indicate that the mutant cells were more sensitive to zinc stress and seemed to have more oxidative intracellular environment than wild type cells. Our results support the idea that superoxide dismutase is an important factor for zinc detoxification in eukaryotes. (© 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

9th Eurasia Conference on Chemical Sciences -- SEP 09-13, 2006 -- Antalya, TURKEY, WOS: 000251582100017, The hexane extracts of eight Cephalaria (Dipsacaceae) species, which were collected from southwestern Anatolia, were obtained by Soxhlet apparatus. The fatty acids were derived to methyl esters and determined by gas chromatography/flame ionization detector (GC/FID) and gas chromatography/mass spectrometry (GC/MS) systems. The dominant fatty acid components and maximum percentages were detected as myristic [in C. joppica (17.48 %)], palmitic [in C. cilicica, C. elmaliensis, C. isaurica, C. scoparia (19.51 %), and C. gazipashaensis], linoleic [in C. joppica (33.02 %), C. elmaliensis, C. dipsacoides, and C. gazipashaensis], (x-linolenic (ALA) [in C. cilicica, C. elmaliensis, C. isaurica, C. scoparia, C. lycica, and C. gazipashaensis (47.95 %)] and oleic [in C. isaurica and C. dipsacoides (40.66 %)] acids. The antioxidant activity of all hexane extracts was evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH), ferric thiocyanate (FTC), and thiobarbituric acid (TBA) methods. The results indicate that hexane extracts of Cephalaria species possess considerable antioxidant activity. The highest radical scavenging activity was detected in C. isaurica (IC50 = 741 mu g/mL). The most effective species on lipid peroxidation are C. lycica and C. gazipashaensis in FTC and TBA assays, respectively. This study reveals that Cephalaria species are attractive sources of fatty acid components, especially the essential ones, as well as of effective natural antioxidants., IUPAC

Geranylgeranyl pyrophosphate synthase (GGPPS) is a key enzyme in carotenoid biosynthetic pathway, catalyzing the synthesis of its C20 precursor. The gene encoding GGPPS from bell pepper (Capsicum annuum) has been cloned in Schizosaccharomyces pombe and its heterologous expression analyzed in this model organism for eukaryotes. We aim to partially direct the ergosterol pathway to carotenoid biosynthesis in this yeast. This yeast is not able to synthesize carotenoids naturally, but it produces farnesyl pyrophosphate (FPP), an intermediate metabolite for both ergosterol and geranylgeranyl pyrophosphate (GGPP) biosynthesis. S. pombe transformants carrying the pepper GGPPS cDNA were named KTG. In one of the transformant (KTG1), heterologous expression of GGPPS gene has been analyzed. The GGPPS gene was shown by dot and Northern hybridization to be successfully transcribed. When water soluble proteins isolated from the transformant cells have been analyzed on SDS-denaturating gels, a ~37 kDaprotein band...

The gua1 gene encoding inosine monophosphate dehydrogenase (IMPDH), which catalyses the first step in de novo biosynthesis of guanosine monophosphate (GMP), was cloned in the yeast Schizosaccharomyces pombe by functional complementation of a gua1ura4-D18 mutant strain from a S. pombe DNA genomic library. Complementation analysis revealed a 1.2 kb fragment which segregation analysis confirmed did not code for a suppressor gene. Only 446 nucleotides of the gua1 gene encoding the IMPDH C-terminal residues were found within this 1.2 kb sequence (GenBank, AJ293460). The comparison of this wild-type fragment with the same fragment from the gua1ura4-D18 mutant revealed that there was a point mutation at position 1261 (guanine -> adenine) from the 5' end, corresponding to the amino acid residue 421 (glycine -> serine) of the enzyme. Dot and Northern analyses showed that the gua1 gene was expressed in transformants as well as in the wild-type and the gua1ura4-D18 mutant, but enzyme activity was only detected in wild-type and transformant cells. It seems likely that a 446 bp fragment from the 3' end of the gua1 gene abolished the point mutation in the mutant strain, suggesting that this fragment participates in the sequences encoding the active domain of IMPDH in S. pombe.

Extracts from the aerial parts of Sanicula europaea L. were investigated for their anti-HN activity, and the 50% ethanolic extract was shown to exhibit the highest activity. A new triterpene saponin glycoside, 21 beta-(angeloyloxy)-3-O-[beta-D-arabinopyranosyl(1-->4)-beta-D-glucopyranosyl(1-->3)-beta-D-glucuronopyranosyl propyl ester]-3 beta,15,16,22 alpha,28 beta-pentahydroxy-Delta(12)-oleanene, saniculoside N (1), in addition to the known phenolic acids, rosmarinic acid (2), and caffeic acid(3) were isolated as major components. Rosmarinic acid was established as the principal active substance.

Metal-based compounds represent promising anticancer therapeutic agents. In this study, the mechanism of action of a novel metal-based drug, a palladium(II) (Pd) complex ([PdCl(terpy)](sac)2H(2)O, terpy=2,2':6',2 ''-terpyridine and sac=saccharinate), was elucidated. The tested compound induced cytotoxicity in nine different human cancer cell lines that originated from various organs, suggesting a broad spectrum of activity. The IC50 values were significantly higher for noncancerous cells when compared with cancer cells. We found that cells treated with the Pd(II) complex exhibited increased caspase 3/7 activities and condensed/fragmented nuclei, as demonstrated by nuclear staining and DNA laddering. Morphological features, such as cellular shrinkage and blebbing, were also observed, indicating that apoptosis was the primary mechanism of cell death. Pd(II) treatment induced DNA double-stranded breaks both in vitro and in vivo, potentially accounting for the source of stress in these cells. Although caspase 3/7 activities were elevated after Pd(II) treatment, silencing or using inhibitors of caspase 3 did not block apoptosis. Other molecules that could potentially play a role in Pd(II)-induced apoptosis, such as p53 and Bax, were also tested using silencing technology. However, none of these proteins were essential for cell death, indicating either that these molecules do not participate in Pd(II)-induced apoptosis or that other pathways were activated in their absence. Hence, this new molecule might represent a promising anticancer agent that exhibits cytotoxicity in p53-mutant, Bax-mutant, and/or caspase 3-mutant cancer cells. (C) 2013 Wolters Kluwer Health vertical bar Lippincott Williams & Wilkins.

Hereditary angio-edema (HAE), characterized by recurrent episodes of angioedema involving the skin and the mucosa of the upper respiratory or the gastrointestinal tracts, results from heterozygosity for deficiency of the serine proteinase inhibitor (serpin), C1 inhibitor (C1-INH).In this study, serum inflammatory cytokine levels and circulating endothelial cells collected from HAE patients during both acute attacks and asymptomatic periods were evaluated.Twenty-four patients with Type I and 1 patient with Type II HAE in an asymptomatic period (Group I), 8 patients with Type I HAE during a mild to moderate acute attack (Group II) and 20 healthy subjects (13 females, mean age: 32.1±8.2years) were included. Serum IL-6, IL-8, IL-1β, TNF-α, vascular endothelial growth factor (VEGF) and endothelial nitric oxide synthase (eNOS) levels were detected by ELISA. Circulating endothelial cells (CECs) and circulating endothelial progenitors (CEPs) were evaluated using Fluorescence Activated Cell Sorting (FACS).Serum eNOS levels of HAE patients were significantly higher than healthy subjects (p0.006) while mean TNF-α levels in Group I were slightly lower (p0.03) than Group II. There were no differences in terms of other inflammatory cytokines between the control subjects and HAE patients who were either in an asymptomatic period or experiencing an acute attack. CECs and CEPs were also similar.These results suggest that an inflammatory response is not necessary to trigger HAE attacks. On the other hand, increased eNOS levels might reflect a sustained hyperpermeability state in HAE patients.

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Various extracts from the leaves of mistletoe (Viscum album L. ssp. album) were investigated for their antiviral activity on human parainfluenza virus type 2 (HPIV-2) growth in Vero cells. Plant extracts were prepared using distilled water, 50% ethanol, petroleum ether, chloroform and acetone. The 50% effective dose (ED(50)) of aqueous extract for HPIV-2 replication was 0.53 +/- 0.12 micro g/mL, and the antiviral index (AI), which was based on the ratio of the 50% inhibitory concentration (CD(50)) for host cell viability to the ED(50) for parainfluenza virus replication, was 10.05. The aqueous extract was found to be the most selective inhibitor. Furthermore, the aqueous extract at a concentration of 1 micro g/mL was found to inhibit HPIV-2 replication and the virus production was suppressed to more than 99% without any toxic effect on host cells. The chloroform extract was also found to be moderately active. In an effort to further analyse the mechanism of antiviral activity, the effectiveness of the aqueous extract on different steps of virus replication was examined. The antiviral activity could neither be attributed to the direct inactivation of the HPIV-2 nor to the inhibition of adsorption to Vero cells. The active aqueous extract has shown a dose-dependent antiviral activity on virus replication.

Viscum albumL. is a semiparasitic plant grown on trees and widely used for the treatment of many diseases in traditional and complementary therapy. It is well known that some activities ofViscum albumextracts are varied depending on the host trees, such as antioxidant, apoptosis-inducing, anticancer activities of the plant. The aim of the present study is to examine the comparative effects of methanolic extracts ofV. albumgrown on three different host trees (locust tree, lime tree, and hedge maple tree) on H2O2-induced DNA damage in HeLa cells. Oxidative damage in mitochondrial DNA and two nuclear regions was assessed by QPCR assay. The cells were pretreated with methanolic extracts (10 μg/mL) for 48 h, followed by the treatment with 750 μM H2O2for 1 hour. DNA damage was significantly induced by H2O2while it was inhibited byV. albumextracts. All extracts completely protected against nuclear DNA damage. While the extract from lime tree or white locust tree entirely inhibited mitochondrial DNA damage, that from hedge maple tree inhibited by only 50%. These results suggest that methanolic extracts ofV. albumcan prevent oxidative DNA damage, and the activity is dependent on the host tree.

Olive oil contains several phenolic compounds possessing antioxidant activity. The aim of this study was to investigate the protective effects of olive oil phenolic extract (OOPE) and one of its constituents, gallic acid (GA) against H(2)O(2)-induced oxidative stress and apoptotic cell death in HeLa cells, a model for human epithelial cells.The cells were pretreated with nontoxic doses of OOPE or GA for 4, 24 and 48 h, and the intracellular reactive oxygen species (ROS) level was determined, before and after oxidative stress induction with H(2)O(2). As an indicator of apoptosis, caspase 9 activity was measured.All pretreatments reduced ROS generation. Four hour incubation with OOPE or GA completely inhibited ROS generation. Increases in caspase 9 activity by OOPE and GA pretreatment under harsh stress conditions were inhibited 92 and 67.8%, respectively.These results suggest that OOPE and GA act as powerful antioxidants against oxidative stress and exert anti-apoptotic effects.

The invertase mutant defective in the glucose signaling pathway of Schizosaccharomyces pombe (ird11) is resistant to glucose repression. This mutant is able to consume sucrose alongside glucose and grows in glucose-containing media with a generation time close to that of the wild type. Intracellular oxidation, protein carbonyl, and reduced glutathione levels and catalase, superoxide dismutase, and glutathione peroxidase activity were investigated in ird11, to determine the relationship between oxidative stress response and glucose signaling. The expression profiles of some genes involved in regulation of glucose repression (fbp1, fructose-1,6-bis-phosphatase; hxk2, hexokinase) and stress response (atf1 and pap1 transcription factors; ctt1, catalase; sod1, Cu,Zn superoxide dismutase) were analyzed using the quantitative real-time PCR technique. Oxidative stress response in ird11 seems to be affected by glucose signaling in a manner different from that caused by glucose deprivation.

Chemical constituents of the aerial parts of four endemic Cephalaria species (Dipsacaceae) in Anatolia, C. scoparia, C. paphlagonica, C. isaurica and C. stellipilis, were investigated. Two new triterpene glycosides from C. scoparia, namely scoposides F (1) and G (2), along with 11 known compounds including phenolic, flavonoid, iridoid and triterpene-type glycosides were isolated from these species. The structures of all compounds were elucidated on the basis of spectroscopic data and chemical evidence. Antioxidant activity of the compounds was tested with CUPRAC method for the first time in addition to the DPPH by using trolox and ascorbic acid as reference antioxidants, respectively. The antimicrobial activity was assessed by MIC assay and the results were given as MIC (mu g/mL) values. New triterpene glycosides scoposides F-G (1 and 2) and the compounds paphlagonoside A (6), paphlagonoside B (7) and isacoside (9), which were not tested previously, exhibited as strong antibacterial activity against Gram (+) and Gram ( ) bacteria. (C) 2011 Phytochemical Society of Europe. Published by Elsevier B. V. All rights reserved.

The objectives of this study were to examine the free radical scavenging activity and the protective effects against macromolecular oxidation as well as the cytotoxic activity of Aphanes arvensis aqueous and methanolic extracts. Free radical scavenging activity was determined by DPPH method. The methanolic extract showed a scavenging activity nearly equivalent to Trolox and Vitamin C and has an IC(50) value of 4.54 microg/mL. Total antioxidant capacity was determined by CUPRAC method. The antioxidant capacity of aqueous and methanolic extract was 0.792 and 1.550 mmol TE/g DWE, respectively. The protective effect of A. arvensis extracts against lipid peroxidation was evaluated using a liposome oxidation system. The methanolic extract was more active than the aqueous extract. The aqueous extract possessed protective effect against protein oxidation in a dose dependent manner. Both extracts showed inhibitory effect on DNA oxidation as measured by plasmid relaxation assay. Results presented here indicate that A. arvensis possess strong antioxidant activity and protective effects with very little cytotoxic effect, and they can therefore be used as a natural additive in food, cosmetic and pharmaceutical industries.

Arda, N. and Sayar, A.A., 1992. Liquid-liquid equilibrium of water + 2-propanol+ 2,2,4-trimethylpentane ternary at 293.2 ± 0.1 K. Fluid Phase Equilibria, 73: 129-138. Tie-lines and the isothermal solubility curve of water + 2-propanol + 2,2,4-trimethylpentane were experimentally determined at 293.2 ± 0.1 K. The reliability of tie-line measurements was checked via Campbell's method and the correlation using Hand's equation. Tie-lines were predicted from the liquid-liquid equilibrium data UNIFAC model. The extracting capability of 2,2,4-trimethylpentane for separating 2-propanol from its aqueous solutions was investigated in terms of distribution coefficients, separation factors and selectivity.

Solubility curve and tie-line measurements for (water + tetrahydrofuran + 1-methylcyclohexanol) and (water + tetrahydrofuran + 2-methylbutyl ethanoate) at the temperature (293.16±0.30)K and the pressure (101.325±0.070)kPa are reported. Tie-lines were correlated using a Hand equation. Consistency of our results was ascertained through an Othmer-Tobias correlation. Distribution coefficients of tetrahydrofuran and water from aqueous solutions to selected high-boiling solvents were determined and separation factors were evaluated. It is concluded that the high-boiling solvent, 2-methylbutyl ethanoate, may be considered a separating agent for dilute aqueous tetrahydrofuran solutions.

A simple and rapid technique for the determination of furfural in the culture medium, which contains this compound as a degradation product of xylose at autoclave conditions, was evaluated. Besides, the growth and the furfural metabolism of the fission yeast Schizosaccharomyces pombe has been monitored using spectral methods.

Zinc is a metal which is a cofactor in many enzymes and a structural element in zinc finger motifs those are important in relation between DNA and regulator proteins. Little is known about uptake, distribution, toxicity and detoxification of zinc ions in cells. In this study, zinc toxicity and detoxification levels have been compared in wild type and Cu/Zn superoxide dismutase mutant (sod1Delta) cells of the fission yeast Schizosaccharomyces pombe. We evaluated the toxic levels of zinc, total zinc content, lipid peroxidation levels and catalase activities for both strains which were grown in medium containing different concentrations of zinc. sod1Delta mutant showed important growth retardation and has higher lipid peroxidation and catalase activities than wild type. Cu/Zn superoxide dismutase (SOD1) activity of wild type cells was markedly increased when they were treated with elevated levels of zinc. SOD1 mRNA level also significantly increased when the cells treated with higher concentrations of zinc. These results indicate that the mutant cells were more sensitive to zinc stress and seemed to have more oxidative intracellular environment than wild type cells. Our results support the idea that superoxide dismutase is an important factor for zinc detoxification in eukaryotes.