Bente Gammelgaard
Lektor
E-mail: bg(at)farma.ku.dk
Telefon: 35 33 64 15
Rum: 13/510

Selenium metabolism
Selenium is an essential element with cancer protective effects. It is constantly debated if populations should be supplied with selenium and if so – which chemical form of selenium is the more efficient as it has been shown that the cancer cells react differently to different selenium compounds. However, the metabolism of different selenium compounds and the influence of metabolism on the cancer protective effect are still not completely understood.
The aim of this ongoing project is to elucidate how selenium is metabolised by healthy individuals as well as cancer cells and elucidate the influence of metabolism on the cancer protective effects. The thesis projects may involve sample preparation, separation and identification of metabolites in plasma, tissue samples or cell lines. The applied methods will be HPLC in combination with MS like ICP-MS and ESI-MS.
Supervisor: Bente Gammelgaard (Max. 2 students)

Characterisation of the interaction of platinum-based drugs with cell membrane transporters
Cisplatin is one of the most widely used anticancer drugs but the applicability of the drug is limited to a relative narrow range of tumour types. Numerous platinum-based drug candidates have been examined with the purpose to minimize side effects and avoid drug resistance. The aim of this project is to improve our knowledge on the resistance mechanism of platinum compounds using selected platinum drug compounds in an in vitro model. The strategy is to use Caco-2 cell cultures as a model for the cancer cells and use competition assays to investigate the role of absorptive/exorptive membrane transporters in development of drug resistance.
Supervisors: Bente Gammelgaard and Bente Steffansen

External projects:
It is also possible to do your thesis work outside FARMA in an industrial or public sector laboratory. Some laboratories may prefer that students spend more than 6 month on the project.

The Department of Forensic Medicine:  Development of new methods for the analysis of drugs and drugs of abuse in biological matrices
The Forensic Chemistry section in Copenhagen has the latest technology in analytical chemistry, and is determined to continuously develop and validate new and improved methods for analysis of pharmaceuticals and drugs of abuse in biological matrices.
Fully automated robotic solid phase extraction (SPE) is frequently used on biological samples, LC-TOF-MS is used for qualitative analysis, and LC-MS/MS is used for quantitative analysis.
Contact: Bente Gammelgaard, external supervisor Sys Stybe Johansen.

Novozymes: Characterisation of peptides and proteins.
Peptide and protein molecules are covalently modified during production and storage and a large number of different modifications may be identified of which some may affect the product stability and performance. The product related impurities are usually characterised by chromatography using several different detection methods like mass spectrometry, UV and pulsed amperometric detection.
The aim of this project is to develop and optimise a generic method for detection of modified amino acid residues. During the project you will use techniques like UPLC, high resolution MS and advanced software tools and get a broad introduction to analytical protein chemistry.
Contact: Bente Gammelgaard, external supervisor: Ulrik Sidenius.

Neurosearch: Microsomes from various tissues
The metabolic stability of new chemical entities is often estimated in microsomes isolated from the liver. However, it has been shown that some compounds, e.g. terfenadine, are metabolised very fast by the metabolising enzymes in the gut wall. Furthermore, it has been shown that the phase II glucuronidation of raloxifene is faster in human intestinal microsomes when compared with liver microsomes. Therefore, lead compounds should be evaluated in a metabolic stability assay, which uses microsomes from other tissues.
The aim of the project is to set-up an assay for evaluation metabolic stability of a range of compounds in intestinal and kidney microsomes isolated from various species. Furthermore, the aim is to set-up an assay for phase II glucuronidation of new chemical entities in intestinal and kidney microsomes.
Contact: Bente Gammelgaard, external supervisor: Kim G. Madsen

Neurosearch: Electrochemical generation of iminium ions and trapping of those
The formation of reactive drug metabolites has been associated with observed toxicity for some compounds. New chemical entities are typically screened for their ability to form conjugates with glutathione (an endogenous tripeptide), which is an indicator for the presence of reactive metabolites. However, not all reactive metabolites can react with glutathione and iminium ions are such type of reactive metabolite. Iminum ions can react with cyanide and thus, identification of cyanide conjugates are an indicator for reactive metabolites. However, it has never been shown which biologic nucleophile, the cyanide ion mimics.
The aim of the project is to set up an electrochemical method for generating reactive iminium ions and to react those with cyanide and biological relevant nucleophiles. Furthermore, the results should be compared with existing in vitro metabolic assays (e.g. by incubation in liver microsomes).
Contact: Bente Gammelgaard, external supervisor: Kim G. Madsen.

Neurosearch: Alternative metabolism reaction
One of the early screening assays in modern drug discovery is metabolic stability in liver microsomes from various species. Only compounds with a reasonable metabolic stability are taken into further development. Hereby, other elimination mechanisms might be relevant for the excretion of the drug and such mechanisms are typically less well-described. Examples of such are the metabolism by either aldehyde oxidase and xanthine oxidase both found in the liver cytosol but also in other tissues. The aim of the project is to set-up assays for other phase I metabolising enzymes.
Contact: Bente Gammelgaard, external supervisor: Kim G. Madsen.

Lundbeck A/S: Immaging mass spectrometry
MALDI-imaging is a powerful technique to visualize the distribution of drugs and metabolites in biological tissues. Over the years there has been increased interest in the analysis of spatial distribution of small molecules in tissues for drug discovery applications, disease diagnosis or biomarker discovery. Especially, the localization or spatial identification of dosed drugs and its metabolites may be critical to understand the mechanism of target-organ toxicity and its relevance to clinical safety. The MALDI-mass spectrometry signal may be obtained directly from tissue sections. The resulting three-dimensional image may become very useful for the investigation of localization of dosed drugs and its metabolites in tissues. However, the technique is highly dependant on a correct preparation of the tissues with matrix modifiers. This project will focus on how to prepare samples with different types of matrix modifiers and solvents in order to obtain the best structural and spatial information for drugs and metabolites in tissue cross-sections
Contact: Bente Gammelgaard, external supervisor: Lars Bendahl

The National Research Centre for the Working Environment:  Biological indicators and stress
Our main purpose is to understand the bodily reactions to work related psychosocial stressors and the influence on health and well-being. The project will establish and evaluate a LC-MS/MS method for determinations of melatonin, cortisol and testosterone in blood. Another project is to establish and evaluate a LC-MS/MS method for determination of catecholamines in urine. The master project will be closely related on-going scientific and will probably also include testing in real samples.
Contact: Bente Gammelgaard, external supervisors: Åse Marie Hansen, Anne Helene Garde.

The National Research Centre for the Working Environment:
Biological indicators in saliva
Our main purpose is to understand the bodily reactions to work related psychosocial stressors and the influence on health and well-being. What can we measure in saliva? To establish methods on LC-MS/MS systems such as Q-TOF and triple quad for qualitative characterization of components in natural saliva samples.
Contact: Bente Gammelgaard, external supervisors: Åse Marie Hansen, Per Axel Clausen.

The National Research Centre for the Working Environment:
Toxicity of carbon nanotubes (CNTs)
CNTs are a large group of compounds which exclusively consists of carbon. CNTs are hollow tubes built of carbon atoms arranged in hexagons. They have diameters from 1 nm to several nm and lengths of several µm. They have tensile strength of 10-100 times the strongest steel and they may be metallic or semiconducting. They are predicted a great future e.g. as molecular fibres in polymer composite materials which are used in wind turbine blades and aeroplanes. However, CNTs have shown to possess different toxic effect such as cytotoxicity, genotoxicity, and carcinogenicity. Some researchers have suggested that the observed cytotoxicity may be due simply to removal of nutrients from the cell media by adsorption and in this way reduces the growth of the cells. This is the hypothesis we want to test. For this purpose we have to establish methods for analysis of vitamins and possibly amino acids in cell media by HPLC and mass spectrometry (MS). The project is related to on-going research projects.
Contact: Bente Gammelgaard, external supervisors: Per Axel Clausen, Asger Nørgaard.