Pramanicin-a induces apoptosis in hct116 colon carcinoma cells: activation of jnk, p38, erk1/2 and induction of oxidative stress

Pramanicin (PMC) is a novel anti-fungal agent. In this study, among eight analogues screened by MTT cell proliferation assay for their potential cytotoxic effect on HCT116 colon cancer cells, Pramanicin-A and Pramanicin-F were found to be the most effective candidates at the concentration of 25uM and 75uM respectively. Flow-cytometric analyses with Annexin-V staining and M30 apoptosense eliza assay confirmed that PMC-A is a more effective apoptotic agent compared to PMC-F, thus PMC-A was selected for further studies. Moreover, no difference in cytotoxicity was observerved in puma and bax defficient cell lines, therefore further studies were conducted with HCT116 wild type cells. In order to get insight into the mechanism of apoptotic response by PMC-A, we followed MAP kinase pathways with using specific MAPK antibodies and inhibitors. Our immunoblotting data reveals that PMC-A induced the activation/phosphorilation of c-jun terminal kinase (JNK), p38 and extracellular signal-regulated kinases (ERK1/2) in different time kinetics. Inhibition of caspase-3, and caspase-9 with their specific inhibitors prevent apoptosis. Interestingly inhibition of JNK and p38 activations/phosphorilations potentiated the apoptotic response. These data indicate that PMC-A induced apoptosis is mediated by caspase dependent pathways, activation of JNK and p38 but not ERK 1/2 may have a pro-survival role.Finally our data from flow-cytometric and flourometric analyses with D2CDF-DA staining revealed induction of reactive oxygen species (ROS) acting as second messengers which may activate MAPK signaling pathways as well as other signaling pathways in apoptotic response of cells to PMC-A at early (1h, 2h) and late (24h) time points

Nitrogen source, an important determinant of fatty acid accumulation and profile in scenedesmus obliquus

The potential of algae-based fuel technologies for manufacturing renewable biofuels has been attracting interest from the scientific community. Biomass productivity and cellular lipid content are important parameters affecting the feasibility of using algae oil for biodiesel production. This study compares utilization of NaNO3, NH4Cland urea as different nitrogen sources in terms of their effects on biomass productivity, fatty acid profile and accumulation in Scenedesmus obliquus. Cellular lipid accumulation was analyzed by gravimetric, fluorometric, and flow-cytometric methods, besides collecting spectrophotometric data for biomass productivity analysis. In addition, fatty acid profiles were compared by using gas chromatography–mass spectrometry. The alga can utilize all tested nitrogen sources successfully however growth rates demonstrate differences. Gravimetric lipid content analysisshowed approximately a 1.5-fold increase in total lipid accumulation under NH4Cl regime when compared tothat of NaNO3 and a 2-fold increase when compared to that of urea at the end of ten days cultivation course. Fatty acid profiles under different nitrogen regimes present variations especially under NH4Cl regime. Moreover,all lipid extracts mostly consist of saturated, straight- and branched-chain hydrocarbons of different chain lengths ranging from C16–C20 which grant a suitable profile for biodiesel production. Scenedesmus obliquus is a suitable species for biodiesel production. The results obtained from this study provide a better understanding of cultivation characteristics of this important species and support potential, future biodiesel production

Oxidative stress is a mediator for increased lipid accumulation in a newly isolated Dunaliella salina strain

Green algae offer sustainable, clean and eco-friendly energy resource. However, production efficiency needs to be improved. Increasing cellular lipid levels by nitrogen depletion is one of the most studied strategies. Despite this, the underlying physiological and biochemical mechanisms of this response have not been well defined. Algae species adapted to hypersaline conditions can be cultivated in salty waters which are not useful for agriculture or consumption. Due to their inherent extreme cultivation conditions, use of hypersaline algae species is better suited for avoiding culture contamination issues. In this study, we identified a new halophilic Dunaliella salina strain by using 18S ribosomal RNA gene sequencing. We found that growth and biomass productivities of this strain were directly related to nitrogen levels, as the highest biomass concentration under 0.05 mM or 5 mM nitrogen regimes were 495 mg/l and 1409 mg/l, respectively. We also confirmed that nitrogen limitation increased cellular lipid content up to 35% under 0.05 mM nitrogen concentration. In order to gain insight into the mechanisms of this phenomenon, we applied fluorometric, flow cytometric and spectrophotometric methods to measure oxidative stress and enzymatic defence mechanisms. Under nitrogen depleted cultivation conditions, we observed increased lipid peroxidation by measuring an important oxidative stress marker, malondialdehyde and enhanced activation of catalase, ascorbate peroxidase and superoxide dismutase antioxidant enzymes. These observations indicated that oxidative stress is accompanied by increased lipid content in the green alga. In addition, we also showed that at optimum cultivation conditions, inducing oxidative stress by application of exogenous H2O2 leads to increased cellular lipid content up to 44% when compared with non-treated control groups. Our results support that oxidative stress and lipid overproduction are linked. Importantly, these results also suggest that oxidative stress mediates lipid accumulation. Understanding such relationships may provide guidance for efficient production of algal biodiesels

Chemical composition and antimicrobial activity of Foeniculum vulgare Mill. essential oil

Foeniculum vulgare Mill. (fennel) is a member of the Apiaceae plant family and is used as an antiinflammatory, analgesic, carminative, diuretic, and antispasmodic agent. Nowadays, there is a growing interest in the antioxidant potential and antimicrobial activities of fennel fruit extracts and essential oil [1]. There are many reports on the essential-oil composition of F. vulgare. Previously, the composition and significant antimicrobial activity of the essential oil of F. vulgare from Pakistan was reported with trans-anethole (70.1%) as the main compound [2]. An essential oil of the fruits of F. vulgare, containing trans-anethole (68.5%) and estragole (10.4%), showed antibacterial activity against Staphylococcus albus, Bacillus subtilis, Salmonella typhimurium, Shigella dysenteriae, and Escherichia coli [3]. In the current study, the essential oil of the aerial parts of F. vulgare was obtained by hydrodistillation (3 h). The essential-oil composition was analyzed by means of gas chromatography-mass spectrometry (GC-MS). The main components of the essential oil from the aerial parts were estragole (33.6%), limonene (24.7%), and α-pinene (19.1%). Additionally, the antimicrobial activity of the essential oil was investigated against Gram-negative (Escherichia coli DH5α) and Gram-positive (Staphylococcus aureus) bacteria. The essential oil showed a growth inhibitory activity against E. coli DH5α (69.3%), tested at 80 mg/mL. However, no activity of the oil was detected in the case of Gram-positive bacteria

Chemical composition and antimicrobial activity of Glebionis coronaria (L.) Cass. ex Spach essential oil

Glebionis coronaria (L.) Cass. ex Spach (syn. Chrysanthemum coronarium L.) is a member of the Asteraceae plant family which has extensive edible, folk medicinal, insecticidal uses. Previously, an essential oil of flowerheads of C. coronarium from Spain was reported to contain camphor (29.2%), α-pinene (14.8%), β-pinene (9.5%), and lyratyl acetate (9.8%). The oil was shown to possess significant antifungal activity [1]. Also, the essential-oil composition and antimicrobial properties of C. coronarium from Ukraine were investigated. The major constituents were found to be chrysanthemyl acetate (24.4%), chrysanthemol (21.8%), chrysanthenyl acetate (7.6%), camphor (7.3%), β-farnesene (5.9%), and α-bisabolol (5.6%). An ethanolic extract of the plant showed antimicrobial activity against Gram-positive (S. aureus) bacteria. [2]. Biological activities of the essential oil of C. coronarium from Jordan were also reported. The essential oil showed a significant antimicrobial activity against Gram-positive bacteria. Also, the oil showed moderate antioxidant activity, weak acetylcholinesterase-inhibitory and potent antiproliferative activities [3]. In the current study, the essential oil of the aerial parts of G. coronaria was obtained by hydrodistillation (3 h). The essential-oil composition was analyzed by means of gas chromatography-mass spectrometry (GC-MS). The main components of the essential oil from the aerial parts were capillene (54.5%) and caryophyllene oxide (9.8%). The current composition differed from the previously reported ones. Thus, G. coronaria originating from Istanbul belongs to a new chemotype of this species. Additionally, the antimicrobial activity of the oil was investigated against Gram-negative (Escherichia coli DH5α) and Gram-positive (Staphylococcus aureus) bacteria. The essential oil showed a growth-inhibitory activity against E. coli (53.3%) and S. aureus (17.1%), tested at 75 mg/mL. The antimicrobial effects of the essential oil were more pronounced against Gram-negative bacteria

Genetic, physiological and biotechnological assessment of microorganisms for renewable and sustainable energy resource production

The term "algae" defines variety of photosynthetic organisms found throughout the world in various environmental conditions. Algae species are estimated to number in the tens of thousands. Because algae are photosynthetic, naturally able to replicate rapidly and produce high amount of oils, alcohols, and biomass, they have attracted the attention of researchers and industrial producers seeking alternatives to currently used fossil fuels. Algae thrive on organic carbon or CO2, nutrients such as nitrogen, phosphorus and other inorganic substances which enables algae to be used in bioremediation. Growth conditions, nutrients such as carbon and nitrogen, and many other factors affect the algal cell metabolism. Thus, manipulation of different cultivation conditions have been shown successful in increasing algal biomass and lipid productivity in order to substitute petroleum use. Algae biotechnology research goals especially include finding ways to increase the reproductive rate, improve metabolism of inputs, and enhance the production of desired oils, fuel-grade alcohols in useful species. In this thesis, newly isolated halophilic unicellular green algae species are assessed for potential renewable energy resource. Novel strategies for increasing cellular lipid production were established. Exogenous application of oxidative stress by hydrogen peroxide treatment was shown as a novel lipid accumulation inducer. Moreover, increased lipid accumulation response was also observed in heavy metal induced oxidative stress which makes combination of heavy metal bioremediation and oil production possible as a novel algae cultivation strategy. Directed evolution and natural selection strategies were applied to model organism Saccharomyces cerevisiae and Dunaliella salina for revealing underlying biochemical, genetic factors of increased cellular lipid production in order to provide useful strategies for future biofuel production

Rediscovery of historical vitis vinifera varieties from the South Anatolia region by using amplified fragment length polymorphism and simple sequence repeat DNA fingerprinting methods

Anatolia played an important role in the diversification and spread of economically important Vitis vinifera varieties. Although several biodiversity studies have been conducted with local cultivars in different regions of Anatolia, our aim is to gain a better knowledge on the biodiversity of endangered historical Vitis vinifera varieties with potential viticulture characteristics in the North Adana region of South Anatolia. We also demonstrate the genetic relatedness in a selected subset of widely cultivated and commercialized Vitis vinifera collection cultivars obtained from the National Grapevine Germplasm at the Institute of Viticulture, Turkey. In the present study,microsatellites were used for pre-selection of 27 collection varieties among 72accessionsto scale down the sample size.We used amplified fragment length polymorphisms to further determine genetic relatedness among collection and local Vitis vinifera cultivars.The UPGMA cluster and principal component analyses revealed that Saimbeyli local cultivars form a distinct group that is distantly related to a selected subset of Vitis vinifera collection varieties from all over Turkey.To our knowledge, this is the first study conducted with these cultivars. Further preservation and use of these potential viticultural varieties will be helpful to avoid genetic erosion and to facilitate agriculture in the region

Essential oil composition and antibacterial activities of Gypsophila species

Essential oil composition of Gypsophila turcica Hamzaoğlu, Gypsophila pinifolia Boiss. & Hausskn., G. tuberculosa Hub.-Mor., G. eriocalyx Boiss. and G. laricina Schreb. were analyzed by means of gas chromatographymass spectrometry (GC-MS). Thirty six, fourty four, sixty six, forty one and sixty one compounds were identified in the essential oils of G. turcica, G. pinifolia, G. laricina, G. tuberculosa and G. eriocalyx respectively. The major components were determined hentriacontane (12.93 0.4%), 1-octadecanol (8.97 0.1%), hexahydrofarnesyl acetone (6.9 0.09%) and pentacosane (6.63 0.08%) in G. turcica oil, hexadecanoic acid (17.6 0.4%), 1-tetradecanol (7.6 0.1%) and phytol (5.63 0.05%) in G. pinifolia oil, octacosane (6.83%), eicosanal (6.19%), triacontane (6.03%) and heneicosane (5.78%) for G. eriocalyx, hexadecanoic acid (25.3%, 27.0%) and hentriacontane (13.0%, 12.6%) for G. tuberculosa and G. laricina, respectively. Antibacterial activity of G. eriocalyx, G. tuberculosis and G. laricina were investigated against Gram negative (Escherichia coli) and Gram positive (Staphylococcus aureus) bacteria