Muhsin Konuk

Determination of genotoxic effects of copper sulphate and cobalt chloride in Allium cepa root cells by chromosome aberration and comet assays

We used the anaphase-telophase chromosome aberration and comet (Single Cell Gel Electrophoresis, SCGE) assays to evaluate the genotoxic effects of copper sulphate (CS) and cobalt chloride (CC) chemicals prepared in two concentrations (EC(50), 2xEC(50)), using methyl methanesulfonate (MMS) as a positive control and untreated cells as a negative control. In Allium root growth inhibition test, EC(50) values for CS and CC are 1.5 and 5.5 ppm, respectively. Mitotic index (MI) decreased in all concentrations tested of CS and CC compared to the control at each exposure time. The bridge, stickiness, vagrant chromosomes, fragments, c-anaphase and multipolarity chromosome aberrations were observed in anaphase-telophase cells. The total chromosome aberrations were more frequent with an increasing in the exposure time and the concentrations of both chemicals. The genotoxicity of CS and CC in Allium cepa root cells was analyzed using a mild alkaline comet assay at pH 12.3, which allows the detection of single strand breaks. In all the concentrations, CS and CC induced a significant increase (P<0.05) in DNA damage. No significant difference was found between positive control (300+/-5.81) and 3 ppm CS (280+/-4.61). The methods used are applicable for biological monitoring of environmental pollutants.

Toxic chemicals-induced genotoxicity detected by random amplified polymorphic DNA (RAPD) in bean (Phaseolus vulgaris L.) seedlings.

Assessment of genotoxins-induced DNA damage and mutations at molecular level is important in eco-genotoxicology. In this research, RAPD was used to detect DNA damage in the roots and leaves of bean (Phaseolus vulgaris L.) seedlings exposed to toxic chemicals of Hg, B, Cr and Zn (HgCl(2), H(3)BO(3), K(2)Cr(2)O(7) and ZnSO(4)7H(2)O) at concentrations of 150 and 350 ppm for 7 d. Inhibition of shoot and root growth and increase of Hg, B, Cr and Zn element contents in the roots and leaves were observed with an increase in the concentration. For the RAPD analyses, 12 RAPD primers of 60-70% GC content were found to produce unique polymorphic band profiles and subsequently were used to produce a total of 120 bands of 263-3125 bp in the roots and leaves of untreated and treated seedlings. Polymorphisms became evident as disappearance and/or appearance of DNA bands in 150 and 350 ppm treatments compared with untreated control treatments. The DNA changes in RAPD profiles were more in the roots than in the leaves. The highest polymorphism was observed in boric acid treatments among the toxic chemicals. In a dendrogram constructed based on genetic similarity coefficients, the treatments were grouped into three main clusters: (a) root-B-150 ppm treatment grouped alone, (b) root-350 ppm-Hg, B, Cr and Zn treatments clustered together, and (c) the others including untreated control treatments merged together. We concluded that DNA alterations detected by RAPD analysis offered a useful biomarker assay for the evaluation of genotoxic effects of Hg, B, Cr and Zn pollutions on plants.

Testing of the mutagenicity and genotoxicity of metolcarb by using both Ames/Salmonella and Allium test.

Mutagenic and genotoxic effects of metolcarb were investigated by both bacterial reverse mutation assay in Salmonella typhimurium TA97, TA98, TA100 and TA102 strains with or without metabolic activation system (S9) and Allium cepa root meristematic cells, respectively. Metolcarb was dissolved in DMSO in Ames/Salmonella test system. 0.1, 1 and 10 microg/plate doses of metolcarb were found to be mutagenic S. typhimurium TA98 without S9. In Allium root growth inhibition test, EC50 value was determined 200 ppm and 0.5xEC50, EC50 and 2xEC50 concentrations of metolcarb were introduced to onion tuber roots and distilled water used as a negative control. Mitotic index (MI), increased in all concentrations compared to control at each exposure time. While disturbed anaphase-telophase, chromosome laggards, stickiness and bridges were observed in anaphase-telophase cells, pro-metaphase, C-mitosis, polyploidy, binuclear cells and disturbed nucleus were observed in other cells. The results were also analyzed statistically by using SPSS for Windows, Mann-Whitney test and Duncans multiple range tests were performed respectively.

Evaluation of genotoxic and mutagenic effects of aqueous extract from aerial parts of Linaria genistifolia subsp. genistifolia

Genotoxic and mutagenic effects of aqueous extract from aerial parts Linaria genistifolia (L.) Mill. subsp. genistifolia, Plantaginaceae (Lg-ext) were investigated by using both Allium cepa root meristematic cells and bacterial reverse mutation assay in Salmonella typhimurium TA98 and TA100 with or without metabolic activation system (S9), respectively. In Allium root growth inhibition test, EC50 value was determined approximately 15 g/L and 0.5xEC50, EC50 and 2xEC50 concentrations of Lg-ext were introduced to onion tuber roots and distilled water and methyl methane sulfonate (MMS, 10 ppm) used as a negative and positive control, respectively. The characteristic effect caused by tested preparations was an increase of mitotic index (MI) in 7.5 g/L and 15 g/L (except 24 and 96 h) and simultaneous decrease of MI in 30 g/L and in MMS. While stickiness, bridges, chromosome laggards and disturbed anaphase-telophase were observed in anaphase-telophase cells, c-metaphase, pro-metaphase, polyploidy and binuclear cells were observed in other cells. Lg-ext was not found to be mutagenic on S. typhimurium TA 98 and TA100 with or without S9. The results were also analyzed statistically by using SPSS for Windows, and Duncans multiple range tests were performed respectively. These results indicate that Lg-ext exhibits genotoxic activity in A. cepa root meristematic cells but not mutagenic activity in Ames test system

Antioxidant Enzyme and Element Status in Heroin Addiction or Heroin Withdrawal in Rats: Effect of Melatonin and Vitamin E Plus Se

Heroin use, withdrawal syndrome, and heroin-related deaths are still the most serious public health problems. Antioxidants and bio-elements are essential for metabolism in living organisms. To our knowledge, there are no data about the effect of antioxidant therapy on the levels of bio-elements and antioxidant enzymes in the naloxone (NX)-induced heroin withdrawal syndrome. Therefore, in the present study for the first time, we have investigated the role of antioxidant therapy, melatonin, and vitamin E plus Se, on the trace and major elements and antioxidant enzymes in the heroin addiction or heroin withdrawal in rats. Glutathione peroxidase levels were increased and catalase levels were decreased in the all study groups when compared to the sham group. The level of superoxide dismutase (SOD) in the fixed dose of heroin (FDH) given group was lower; however, in the variable doses of heroin (VDH) given group SOD level was higher. Furthermore, in withdrawal syndrome, Fe, Mg, Mn, and Ti levels were diminished and Al, Ca, and Cu levels were increased in the FDH+NX group. Moreover, Mg, Mn, and Se levels were also diminished and Al level was increased in the VDH+NX group. In conclusion, our results obviously indicated that heroin effected both bio-element status and antioxidant enzyme activities and, exogenous melatonin or vE+Se therapy might relieve on the element and antioxidant enzyme the destructive activity caused by heroin.

In vitro propagation of an endangered plant species, Thermopsis turcica (Fabaceae)

This report deals with micropropagation of the critically endangered and endemic Turkish shrub, Thermopsis turcica using callus, root and cotyledonary explants. Callus cultures were initiated from root and cotyledon explants on MS medium supplemented with 0.5–20 µM NAA or 2,4-D. The root explants were found to be better in terms of quick responding and callusing percentages as compared to the cotyledons. Organogenic callus production with adventitious roots and shoots were obtained on MS medium with only NAA. The calli obtained with NAA, root and cotyledonary explants were cultured with BA and kinetin (2–8 µM) alone or in combination with a low level (0.5 µM) of 2,4-D or NAA. The best regeneration of shoots from root explants was observed on hormone-free MS medium. NAA with BA or kinetin in the medium improved shoot induction from the calli obtained with NAA. Maximum percentage of shoots (93.3%), maximum number of shoots (6.2) and maximun length of shoots (8.22 cm) were achieved from cotyledonary explants at 4 µM BA and 0.5 µM NAA. The presence of 0.5 µM or higher levels of 2,4-D in shoot induction medium inhibited the regeneration in T. turcica explants. 83% of in vitro rooting was attained on pulsed-IBA treated shoots. The regenerated plants with well developed shoots and roots were successfully acclimatized. Application of this study’s results has the potential to conserve T. turcica from extinction.

Antifungal Activity and Chemical Composition of Essential Oil of Origanum Hypericifolium

Endemic oreganos, Origanum hypericifolium O. Schwartz and P.H. Davis, essential oil was extracted to exert its biological activity in vitro. Fifteen components in its extracts performed by hydro distillation. The major components in the fruit and flower volatiles of O. hypericifolium were p-cymene (34.33 g/100 g oil), carvacrol (21.76 g/100 g oil), thymol (19.54 g/100 g) and γ-terpinene (13.91 g/100 g oil). The antifungal activity of O. hypericifolium’s oil was evaluated against 14 fungi isolated from hazelnut and walnut. Nuts are capable of harboring toxigenic fungi and the threat of mycotoxin contamination on them exists. Essential oil of O. hypericifolium was found to be active both in contact and headspace assays in vitro producing hyphal growth inhibition. In the contact assay, P. frequentans was found to be the least sensitive species. The more sensitive species were P. castellonense, P. verrucosum. var. cyclopium, C. globosum, and A. kiliense. Their growth was completely (100%) inhibited at days 3 and 6. In the volatile assay, all the mycelial growth of all tested fungi was completely inhibited at day 3. The volatile activity was found to be highly efficient than that of contact activity assay. This could be because of the aromatic contents of Origanum, such as monoterpenes, carvacrol, thymol, and p-cymene.

Ethnobotanical survey of plants used in Afyonkarahisar-Turkey

BackgroundThe traditional knowledge about plants and their uses in Turkey is disappearing in recent years because the new generations of villagers migrate to big cities for a better life. Afyonkarahisar located at the intersection of roads and phytogeographical regions (Mediterranean, Iran-Turan, and Euro-Siberian) has more than 2500 plant species. This richness of plant diversity promotes the indigenous commuity for the traditional use of wild plants. The aim of the study is to show wild plants’ ethnobotanical usages associated with medicinal, food, fodder, and household goods in 31 settlements within the boundaries of Afyonkarahisar province.MethodsThe ethnobotanical data were collected from 46 informants by means of semi-structured interviews from 2012 to 2014. Ethnobotanical uses of plants of the study area were conducted in the vicinity of Afyonkarahisar (5 districts, 8 towns, 15 villages, and 3 neighborhood centers).ResultsOne hundred and thirty plant taxa belonging to 39 families were recorded and collected. Hundred and seventy-eight different uses of these plants were documented and used generally for medicinal (84), food (68), fodder (16), household goods (3), dyes (3), handicrafts (3) and religious (1).ConclusionThis study provides interesting uses of plants in the local community of Afyonkarahisar and its surrounding area, in what purpose they make use of plants, how they make use of them and obtained results will contribute to economy of villagers. Since the local people, especially in villages, are poor and do not have health care, they use the plants to treat illnesses, food, fodder, household goods and other uses (evil eye). Also this study will light the way for posterity for next generations.

Seasonal Variations in Fatty Acid Composition of Different Tissues of Mirror Carp, Cyprinus Carpio, in Orenler Dam Lake, Afyonkarahisar, Turkey

The seasonal variations of fatty acids compositions in the muscle, liver, and ovarian tissues of mirror carp (Cyprinus carpio) living in Örenler Dam Lake were determined by using gas chromatography. The results obtained were subjected to statistical analyses by employing SPSS software and p < 0.05 was accepted as significant value. In the statistical analyses, multiple comparisons tests were performed. The results showed that monounsaturated fatty acids levels were found to be higher than that of polyunsaturated fatty acids and saturated fatty acids in all seasons. Palmitic and stearic acid levels were the highest in ovaries (19.95% in winter and 7.79% in summer, respectively). Oleic and palmitoleic levels were at the highest levels in liver tissues (34.53% in summer and 18.37% in winter, respectively). Docosahexaenoic (C22:6ω3, DHA), eicosapentaenoic (C20:5ω3, EPA), eicosatrienoic acid, and arachidonic (C20:4ω6, AA) levels were at the highest level as 17.27, 2.54, and 8.41% in ovaries in the winter and 6.37% in muscle tissues in the spring, respectively. ω3/ω6 ratios were 0.62–0.98 in muscle, 1.54–2.82 in the liver, and 2.82–3.89 in the ovaries. From the point of total fatty acid variations, the highest variations were observed as follows: muscle in spring (26.73%), liver in winter (33.85%), and ovaries in winter 36.38%.

Biosynthesis of nebularine (purine 9-β-D-ribofuranoside) involves enzymic release of hydroxylamine from adenosine

Abstract Biosynthesis of the antibiotic nebularine (purine-9-β- d -ribofuranoside) by Lepista nebularis and Streptomyces yokosukanensis has been studied and a novel enzymic activity is described which deaminates adenosine to release hydroxylamine. Use of 14 C-labelled nucleosides showed that adenosine was the more immediate precursor of nebularine. That formation of nebularine involves direct incorporation of adenosine and does not involve prior catabolism and re-use of catabolic fragments, was shown by locating 82% of the incorporated radioactivity from [8- 14 C]adenosine in C-8 of nebularine. A crude nebularine-forming enzymic extract was fractionated by (NH 4 ) 2 SO 4 precipitation and the activity recovered in the 100% satn supernatant; it was not sedimented by centrifuging for 90 min at 113 000 g. Further purification was achieved by chromatography of Sephacryl S-200 (173-fold) and on BrCN-activated Sepharose 4B (320-fold). Lability of the enzyme during concentration, by various techniques, obviated sequential use of these steps. Activity was not stimulated by pyridine nucleotides or flavins, and a range of metal ions were without effect. Various purine riboside analogues were not inhibitory, although some end-product inhibition was seen with nebularine. Gel-filtration and SDS-PAGE indicated a M r of 9500–10000 for the enzyme. That hydroxylamine is a product of the catalysed reaction was demonstrated chemically and by MS. Use of [ 15 N-amino]adenosine confirmed that the hydroxylamine originates from the 6-amino group of adenosine. The quantitative relationship between nebularine production and metabolism of adenosine to other compounds was studied. Of the total radioactivity from [8- 14 C]adenosine recovered, 3% was in nebularine. The work describes the first reported natural occurrence, in a free state, of purine and its 5′-ribotide.