Reactions of Chlorambucil and its main metabolite, Phenylacetic Acid Mustard, with 2’-deoxyribonucleosides and Calf Thymus DNA

Chlorambucil is an anticancer agent used in the treatment of a variety of cancers, especially in chronic lymphocytic leukemia, and autoimmune diseases. Nevertheless, chlorambucil is potentially mutagenic, teratogenic and carcinogenic. The high antitumor activity and high toxicity of chlorambucil and its main metabolite, phenylacetic acid mustard, to normal tissues have been known for a long time. Despite this, no detailed chemical data on their reactions with biomolecules in aqueous media have been available. The aim of the work described in this thesis was to analyze reactions of chlorambucil with 2’-deoxyribonucleosides and calf thymus DNA in aqueous buffered solution, at physiological pH, and to identify and characterize all adducts by using modern analyzing methods. Our research was also focused on the reactions of phenylacetic acid mustard with 2’-deoxynucleosides under similar conditions. A review of the literature consisting of general background of nucleic acids, alkylating agents and ultraviolet spectroscopy used to identify the purine and pyrimidine nucleosides, as well as the results from experimental work are presented and discussed in this doctoral thesis.Siirretty Doriast

Precipitation of hydroxyapatite at 37 °C in a meso oscillatory flow reactor operated in batch at constant power density

A meso oscillatory flow reactor (OFR) was successfully applied for the precipitation of hydroxyapatite (HAp) nanoparticles. Mixing efficiency of the mesoreactor operated batchwise in a vertical tube was evaluated at constant power density, by monitoring variation of hue values upon mixing both spatially and temporally. The best operating conditions for fast mixing and a more homogeneous reaction medium were verified for f = 0.83 Hz and x0 = 4.5 mm. HAp precipitation was then carried out under these conditions at 37 °C for different mixing Ca/P molar ratios. HAp nanoparticles with a mean size (d50) of about 67 nm and a narrow size distribution were obtained. Furthermore, the obtained results show the advantages of the meso-OFR over a stirred tank batch reactor due to the production, about four times faster, of smaller and more uniform HAp nanoparticles. © 2013 American Institute of Chemical Engineers AIChE J, 2013This work was supported by the Portuguese Foundation for Science and Technology (SFRH/BD/42992/2008) through the MIT-Portugal Program, Bioengineering Systems Focus Area. The authors are thankful to Dr Jorge Ferreira from LNEG (Laboratorio Nacional de Energia e Geologia, Portugal) for carrying out the X-ray measurements and his help with the interpretation of results

Photo-responsive polymeric structures based on spiropyran

Spiropyrans are one of the most popular classes of photochromic compounds that change their optical and structural properties in response to external inputs such as light, protons and metal ions, making them ideal molecules for the fabrication of multifunctional stimuli-responsive materials. Nowadays, the emphasis in polymeric materials incorporating spiropyran units, focuses on the effectiveness of their reversible response to external photonic stimuli. Photo-control of a range of key characteristics for flow systems, such as wettability, permeability, photo-modulation of flow by photo-actuation of valves, photonic control of uptake and release of guests using films and coatings, and colorimetric sensing of various species, are highlighted and discussed

Surface faceting and reconstruction of ceria nanoparticles

The surface atomic arrangement of metal oxides determines their physical and chemical properties, and the ability to control and optimize structural parameters is of crucial importance for many applications, in particular in heterogeneous catalysis and photocatalysis. Whereas the structures of macroscopic single crystals can be determined with established methods, for nanoparticles (NPs), this is a challenging task. Herein, we describe the use of CO as a probe molecule to determine the structure of the surfaces exposed by rod-shaped ceria NPs. After calibrating the CO stretching frequencies using results obtained for different ceria single-crystal surfaces, we found that the rod-shaped NPs actually restructure and expose {111} nanofacets. This finding has important consequences for understanding the controversial surface chemistry of these catalytically highly active ceria NPs and paves the way for the predictive, rational design of catalytic materials at the nanoscale.Postprint (author's final draft

Disposable electrochromic polyaniline sensor based on a redox response using a conventional camera: A first approach to handheld analysis

We present a disposable optical sensor for Ascorbic Acid (AA). It uses a polyaniline based electrochromic sensing film that undergoes a color change when exposed to solutions of ascorbic acid at pH 3.0. The color is monitored by a conventional digital camera working with the hue (H) color coordinate. The electrochromic film was deposited on an Indium Tin Oxide (ITO) electrode by cyclic voltammetry and then characterized by atomic force microscopy, electrochemical and spectroscopic techniques. An estimation of the initial rate of H, as ΔH/Δt, is used as the analytical parameter and resulted in the following logarithmic relationship: ΔH/Δt = 0.029 log[AA] + 0.14, with a limit of detection of 17 μM. The relative standard deviation when using the same membrane 5 times was 7.4% for the blank, and 2.6% (for n = 3) on exposure to ascorbic acid in 160 μM concentration. The sensor is disposable and its applicability to pharmaceutical analysis was demonstrated. This configuration can be extended for future handheld configurations.We acknowledge financial support from the Junta de Andalucía (Proyecto de Excelencia P10-FQM-5974) and from the Ministerio de Economía y Competitividad (CTQ2013-44545-R). These projects were partially supported by European Regional Development Funds (ERDF)

Comparative cytotoxicity of artemisinin and cisplatin and their interactions with chlorogenic acids in MCF7 breast cancer cells.

In parts of Africa and Asia, self-medication with a hot water infusion of Artemisia annua (Artemisia tea) is a common practice for a number of ailments including malaria and cancer. In our earlier work, such an extract showed better potency than artemisinin alone against both chloroquine-sensitive and -resistant parasites. In this study, in vitro tests of the infusion in MCF7 cells showed high IC50 values (>200 μM). The combination of artemisinin and 3-caffeoylquinic acid (3CA), two major components in the extract, was strongly antagonistic and gave a near total loss of cytotoxicity for artemisinin. We observed that the interaction of 3CAs with another cytotoxic compound, cisplatin, showed potentiation of activity by 2.5-fold. The chelation of cellular iron by 3CA is hypothesized as a possible explanation for the loss of artemisinin activity.This study was funded by the Engineering and Physical Sciences Research Council (EPSRC, UK) and SensaPharm Ltd. via an Industrial CASE PhD studentship. The award was allocated competitively by the Chemistry Innovation Knowledge Transfer Network (CIKTN, UK).This is the final published version. It was originally published by Wiley at http://onlinelibrary.wiley.com/doi/10.1002/cmdc.201402285/abstract

Contrasting cellular uptake pathways for chlorido and iodido iminopyridine ruthenium arene anticancer complexes

The pathways involved in cellular uptake and accumulation of iminopyridine complexes of general formula [Ru(η6-p-cymene)(N,N-dimethyl-N′-[(E)-pyridine-2-ylmethylidene]benzene-1,4-diamine)X]PF6 bearing two different halido ligands X = Cl or I, have been explored. The ratio of passive/active cellular accumulation of Ru in A2780 human ovarian cancer cells is compared and contrasted with cisplatin. Also, saturation of cellular uptake, time-dependence of cellular influx/efflux equilibria, together with endocytotic pathways such as caveolae and facilitated diffusion are investigated and discussed. Temperature dependence studies of Ru accumulation in the A2780 cells show that in contrast to cisplatin (CDDP) and chlorido complex 1, which are taken up largely through active transport, the iodido complex 2 enters cells via passive transport. The cellular efflux of Ru is slow (ca. 25% retained after 72 h) and is partially inhibited by verapamil, implicating the P-gp protein in the efflux mechanism. Ouabain inhibition experiments suggest that the cellular uptake of these ruthenium complexes relies at least in part on facilitated diffusion, and in particular is dependent on the membrane potential. In addition the finding that depletion of cellular ATP with antimycin A had little effect on cellular Ru accumulation from iodido complex 2 is consistent with passive diffusion. In contrast, ATP depletion caused a major increase in cellular accumulation of ruthenium from chlorido complex 1

LHC String Phenomenology

We argue that it is possible to address the deeper LHC Inverse Problem, to gain insight into the underlying theory from LHC signatures of new physics. We propose a technique which may allow us to distinguish among, and favor or disfavor, various classes of underlying theoretical constructions using (assumed) new physics signals at the LHC. We think that this can be done with limited data $(5-10 fb^{-1})$, and improved with more data. This is because of two reasons -- a) it is possible in many cases to reliably go from (semi)realistic microscopic string construction to the space of experimental observables, say, LHC signatures. b) The patterns of signatures at the LHC are sensitive to the structure of the underlying theoretical constructions. We illustrate our approach by analyzing two promising classes of string compactifications along with six other string-motivated constructions. Even though these constructions are not complete, they illustrate the point we want to emphasize. We think that using this technique effectively over time can eventually help us to meaningfully connect experimental data to microscopic theory.Comment: 50 Pages, 13 Figures, 3 Tables, v2: minor changes, references adde

Pharmacological Effects of Asiatic acid in Glioblastoma Cells under Hypoxia

Glioblastoma multiforme (GBM) is the most common and malignant primary brain tumor in adults. Despite current treatment options including surgery followed by radiation and chemotherapy with temozolomide (TMZ) and cisplatin, the median survival rate remains low (<16 months). Combined with increasing drug resistance and the inability of some compounds to cross the blood brain barrier (BBB), novel compounds are being sought for the treatment of this disease. Here, we aimed to examine the pharmacological effect of Asiatic acid (AA) in glioblastoma under hypoxia. To investigate the effects of AA on cell viability, proliferation, apoptosis and wound healing, SVG p12 fetal glia and U87-MG grade IV glioblastoma cells were cultured under normoxic (21% O2) and hypoxic (1% O2) conditions. In normoxia, AA reduced cell viability in U87-MG cells in a time and concentration-dependent manner. A significant decrease in viability, compared to cisplatin, was observed following 2hrs of AA treatment with no significant changes in cell proliferation or cell cycle progression observed. Under hypoxia, a significantly greater number of cells underwent apoptosis in comparison to cisplatin. While cisplatin showed a reduction in wound healing in normoxia, a significantly greater reduction was observed following AA treatment. An overall reduction in wound healing was observed under hypoxia. The results of this study show that AA has cytotoxic effects on glioma cell lines and has the potential to become an alternative treatment for glioblastoma