UNESCO Trace element satellite centre at Charles University in Prague
Department of Analytical Chemistry, Faculty of Science,
Albertov 2030, 128 43 Prague 2, CZECH REPUBLIC,
Phone: +420-2-2195 2295, Fax: +420-2-24913538,
E-mail: barek@natur.cuni.cz

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Steering committee
Research Program

Steering committee

Prof. RNDr. J.Barek, PhD., FRSC., C.Chem., EurChem.
Head of UNESCO Trace element satellite centre at Charles University in Prague
Dept.Anal.Chem.,Charles University, Faculty of Science, Albertov 2030, 128 43 Prague 2
CZECH REPUBLIC, Phone: +420-2-2195 2295, Fax: +420-2-24913538,

Prof. Ing. K.Štulík, D.Sc.
Head of Department of Analytical Chemistry
Charles University, Faculty of Science, Albertov 2030, 128 43 Prague 2,
CZECH REPUBLIC, Phone: +420-2-2195 2434, Fax: +420-2-24913538,
E-mail: stulik@natur.cuni.cz

Prof. V.Bencko, M.D., D.Sc.
Head of the Institute of Hygiene and Epidemiology, The First Faculty of Medicine, Charles University,
Studnickova 7, 128 00 Prague 2, CZECH REPUBLIC, Phone: +420-2-292 431,
E-mail: Vladimir.Bencko@lf1.cuni.cz

Prof. O.Topolcan, M.D., PhD.
Head of the 2nd Intern Clinic, Faculty Hospital Plzen, Tr. E.Benese 13, 305 99 Plzen
tel/fax: +420 19 27 38 08, E-mail: topolcan@fnplzen.cz, www.fnplzen.cz

Prof. RNDr. PhMr. R. Kalvoda, D.Sc.
J. Heyrovsky Institute of Physical Chemistry, Academy of Science, Czech Republic, Dolejskova 3, 183 23 Praha 8
tel: +420 2 6605 3875

Research Program

  1. The study of the Influence of trace elements on human fertility
  2. The significance of the assessment of trace elements for the epidemiology, etiopathology and prognosis of breast and colorectal tumours
  3. Investigation of metallothioneins using modern electrochemical methods (differential pulse voltammetry, cyclic voltammetry, adsorptive stripping voltammetry and high performance liquid chromatography with electrochemical detection)
  4. Development of new analytical methods for trace elements
  5. Training researchers from other UNESCO Trace element satellite centres in modern methods of trace analysis, organisation of short and medium length courses and PhD study in trace element analysis
  6. Organisation of visits for the representatives of the UNESCO Trace element satellite centre at Charles University in Prague to the headquarters of the Institute in Lyon, France and to the other UNESCO Trace element satellite centres

1. The objective of the recent investigation is to trace the presence of cadmium and zinc in the organism of sterile women included in an IVF program: in blood and follicular fluid (i.e. in a medium which surrounds the gamete- the oocyte- and to follow up their concentrations in relation to achievement of pregnancy. Cadmium and zinc in blood and follicular fluid is assessed on a mass spectrometer with induction bound plasma as the source of ions (ICP/MS,Varian). The potential cause of fertility disorders in conjunction with toxic trace elements is investigated. In cooperation with the Gynecological-maternity clinic of the 1st Faculty of Medicine of Charles University subject of our present interest is a male fertility in the same context.

2. The significance of the assessment of trace elements and biologically active substances for the epidemiology, etiopathology and prognosis of breast and colorectal tumours

(O. Topolčan, V. Bencko, J. Fínek, R. Pikner, L. Holubec, J. Rameš, I. Němcová, P. Rychlovský, M. Mihaljevič, M. Šťastná, Š. Svobodová)

Today tumour diseases are the second most frequent cause of death on industrially developed countries. In the Czech Republic, breast and colorectal carcinomas are two most widespread types of tumours. In the inception and development of tumour diseases a combination of internal, genetically conditioned and external factors are present. In these factors, apart from exposure to carcinogenic substances, life style plays an important role1. The task of epidemiology is to contribute to the detection of risk factors and, with appropriate intervention, to eliminate them by means of primary prevention, or, at least, to limit their effect in the population. Exposure to trace elements represents one of the important risk factors of the environment. Some trace elements are recognised by IARC among chemicals carcinogenic to human: arsenic, beryllium, cadmium and their compounds and chromium [VI] compounds, nickel compounds2. On the other hand, certain trace elements (e.g. selenium), playing an important role in our nutrition, have been investigated as regards their effect for cancer prevention3. Epidemiological studies have consistently shown that human population having a low selenium intake and correspondingly low plasma or serum selenium levels have an increased incidence of a variety of cancers4. These findings are of great importance since the inhabitants of the Czech Republic have been shown to have one of the lowest selenium status in Europe5,6.

One area of interest in present-day studies of the etiopathogenesis of tumour diseases is an attempt to establish which biologically active substances will facilitate the determination not only of the earliest possible diagnosis but more particularly the prognosis of further development of the tumour disease. Nowadays there are several method used based on histological and histochemical examinations, determining and assessing the dynamics of tumour marker levels in biological fluids and determining of specific cellular proteins and enzymes in the cytosol of tumour cells. Moreover, it has been observed that certain trace element concentrations in biological fluids and cancerous tissues of patients are altered as secondary effects of tumour growth. Knowledge about trace element metabolism and its disturbances in tumor tissue or in the organism carrying a malignancy, might help to elucidate the relation of the elements to cancer and to decide whether these elements could be used as additional biochemical markers for the diagnosis or prognosis of tumour development.

It is known that many trace elements participate in biological processes in the organism, which include the activation or inhibition of enzymatic reactions, competition between elements and metalloproteins for binding positions, and modifications in the permeability of cellular membranes. It can therefore be presumed that these elements might influence carcinogenic processes. There are few studies concerning the determination of trace element levels in blood (urine, hair) of breast or colorectal cancer patients. Most of them deal with zinc, copper and selenium levels because of the role of these elements in the antioxidant defence system of the organism. The results are contradictory and not consistent.

Some authors7-10 found the mean serum copper level and the mean Cu/Zn ratio in patients with breast cancer significantly higher than the control group, and the mean serum zinc level in patients significantly lower than the controls. Garofalo et al.11 and Piccinini et al.12 reported that neither the serum Cu nor Zn levels are of value in discriminating between the patients with benign and malign breast cancer and the healthy controls, and in determining the severity of the disease. On the contrary, Cavallo et al.13, in a two-centred study performed in Northern Italy and Southern France observed serum zinc levels to be significantly higher in patients in both centres than in controls, whereas serum copper levels were found to be significantly high in controls in Italy and in patients in France. As for selenium, several authors reported decreased selenium level in patients with cancers breast and colorectal cancers14-16. However, this drop could be interpreted as either the consequence or even the cause of the disease. It could be due to the selenium needs of tumour, to the metabolic consequences of the tumour or even to a low dietary intake of Se.

Multielemental techniques such as neutron activation (NAA) and inductively coupled plasma mass spectroscopy (ICP-MS), make possible the simultaneous determination of ultra-trace quantities of elements in human tissues and body fluids. By using such techniques, it is possible to determine whether the simultaneous monitoring of the less abundant trace metals has diagnostic or prognostic significance. Such systematic studies which include a broad spectrum of trace elements with large patient populations are very scarce. They concern trace element levels in cancerous and normal breast tissue17-19. Again, data reported by different authors are not consistent although they agree as to the significant enhancement of certain elements in the cancerous vs adjacent normal tissue. According to the authors, the elevations of concentrations of elements are probably due to the increased cellular activity in malignant tissue and active enzymatic systems leading to increased amounts of trace elements.

The presented project aims to help to elucidate the role of trace elements status of patients with breast and colorectal carcinomas with respect to their environment and life style. We will investigate trace element levels in blood and cancerous tissues of the patients together with the assessment of life style risk factors (using an epidemiological questionnaire) and the determination of cytosolic and serial examination of tumour markers. In our knowledge, such complex approach has not been undertaken so far and it will consist in the co-operation of experts from different fields - epidemiology, oncology, immunology, analytical chemistry. The project will be the pilot study for further research on the significance of trace elements in carcinogenesis.

During the preliminary study, the sampling protocol will be established, trace elements selected and this study will also lead to the optimisation of the ICP-MS technique. Aside from essential trace elements such as zinc, copper and selenium, we aim to determine elements such as Rb, Sr, Pb, Tl, ..., whose function in the carcinogenesis has not been taken into account so far.

The project is considered to 3 years. The study group will consist of 100 patients with breast cancer and 100 patients with colorectal cancer from Oncological centre of Faculty Hospital Plzeň. In 1st and 2nd year there will be the examination of patients with first diagnosis of tumour. Each patient will undergo:

During the project, patients will be followed in 3-month intervals and there will be the assessment of the same tumour markers and trace elements as during the first examination. Trace elements levels will be correlated with tumour marker levels and with the evolution of the disease and treatment.


  1. Holcátová I., Bencko V.: Environmental epidemiology of malignancies. The Central European perspective. Centr.eur.J.publ.Hlth 6, No 1, 13-17 (1998).
  2. IARC Monographs. Overall Evaluation of Carcinogenicity to Humans. IARC (1998).
  3. Clark L.C., Combs G.F., Turnbull B.W et al.: Effects of selenium supplementation for cancer prevention in patients with carcinoma of the skin. JAMA, 276, No 24, 1957-1963 (1996).
  4. Clark LC. The epidemiology of selenium and cancer. Fed Proc 1985, 44(9): 2584-2589.
  5. Kvíčala J., Zamrazil V., Čeřovská J. et al.: Evaluation of selenium supply and status of inhabitants in three selected rural and urban regions of the Czech Republic. Biol. Trace Elem. Res., 47, 365-375 (1995).
  6. Kvíčala J., Zamrazil V., Čermák S.: Levels of some trace elements in serum of Prague inhabitants measured by INAA. Biol Trace Elem Res ;43-45, 497-502(1994).
  7. Yücel I, Arpaci F, Özet A, Döner B, Karayinoglu T, Sayar A, and Berk Ö. Serum copper and zinc levels and copper/zinc ratio in patients with breast cancer. Biol. Trace Element Res. 1994, 40:31-38.
  8. Gupta SK, Shukla VK, Vaidya MP, Roy SK, and Gupta S. Serum trace elements and Cu/Zn ratio in breast cancer patients. J. Surg. Oncol. 1991, 46:178-182.
  9. Capel ID, Pinnock MH, Williams DC, Hanham IW. The serum levels of some trace and bulk elements in cancer patients. Oncology 1982, 39:38-41.
  10. Borella P, Bargellini A, Caselgrandi E, and Piccinini L. Observations on the use of plasma, hair and tissue to evaluate trace element status in cancer. J. Trace Elem. Med. Biol. 1997, 11: 162-165.
  11. Garofalo JA, Ashiraki H, Menendez-Botet C, Cunningham-Rundles S, Schwartz MK, and Good RA. Serum zinc, copper, and the Cu/Zn ratio in patients with benign and malignant breast lesions. Cancer 1980, 46:2682-2685.
  12. Piccinini L, Borella P, Bargellini A, Medici CI, and Zoboli A. A Case -control study on selenium, zinc, and copper in plasma and hair of subjects affected by breast and lung cancer, Biol. Trace Element Res. 1996, 51:23-30.
  13. Cavallo F, Gerber M, Marubini E, Richardson S, Barbieri A, Costa A, DeCarli A, and Pujol H. Zinc and copper in breast cancer. Cancer 1991, 67(3):738-745.
  14. Psathakis D, Wedemeyer N, Oevermann E, Krug F, Siegers CP, and Bruch HP. Blood selenium and glutathione peroxidase status in patients with colorectal cancer, Dis. Colon Rectum 1998, 41:328-335.
  15. Nelson RL, Davis FG, Sutter E, Kikendall JW, Sobin LH, Milner JA, and Bowen PE. Serum selenium and colonic neoplastic risk, Dis. Colon Rectum 1995, 38: 1306-1310.
  16. Pawlowicz Z, Zachara BA, Trafikowska U, Maciag A, Marchaluk E, Nowicki A. Blood selenium concentrations and glutathione peroxidase activities in patients with breast cancer and with advanced gastrointestinal cancer. J Trace Elem Electrolytes Health Dis 1991, 5(4): 275-277.
  17. Rizk SL and Sky-Peck HH. Comparison between concentrations of trace elements in normal and neoplastic human breast tissue, Cancer Res. 1984, 44:5390-5394.
  18. Garg AN, Singh V, Weginwar RG, and Sagdeo VN. An elemental correlation study in cancerous and normal breast tissue with successive clinical stages by neutron activation analysis. Biol. Trace Element Res. 1994, 46:185-202.
  19. Ng KH, Bradley DA, and Looi LM. Elevated trace element concentrations in malignant breast tissues, Br. J. Radiol. 1997, 70:375-382.

3. Investigation of metallothioneins using modern electrochemical methods

(I. Šestáková)

(differential pulse voltammetry, cyclic voltammetry, adsorptive stripping voltammetry and high performance liquid chromatography with electrochemical detection)

Metallothioneins (MT) are low molecular mass proteins (6-7 kDa), characterised by high cysteine content and ability to bind metals , most important being Zn, Cd, Cu, Hg. Formation of metal - thiolate bonds possessing specific spectroscopic features and arranged in metal- thiolate clusters is important characteristic of metallothioneins, which is studied extensively and summarised in several books and monographs1-5. Besides most frequently studied mammalian MT, metallothioneins isolated from mussels, fungi and plants have been reported. At the end of 1997, more than 170 amino acid sequences of metallothionein from 50 species have become known and analysed5.

Metallothioneins are considered to be important in homeostatic control, metabolism and detoxification of several essential or toxic trace metals. Also, they act as antioxidants and radical scavengers, protecting DNA damage. Several studies also suggest that metallothionein can protect DNA damage caused by metals and other chemicals e.g. chromium, nickel, alkylating agents4-6.. Cd and Zn containing metallothioneins are studied extensively on mercury electrodes7-15 and also on carbon electrodes16.

The aim of the research in this field is:

  1. Use of electrochemical methods for speciation of cations bound to cysteine moiety
  2. estimation of apparent stability constants of complexes in different media (pH, presence of other ions etc).
  3. Combination of electroanalytical methods with separation techniques, e.g. HPLC, capillary electrophoresis.
  4. The investigation of the interactions of MT in systems producing radicals or containing other species inducing damage of DNA.
  5. Environmental monitoring - MT proposed as a tool for trace metal monitoring - marine environment, different types of algae.
  6. Development of new types of electrodes and electrochemical software suitable for electrochemical investigation of MT


  1. M. J. Stillman, C. F. Shaw III, K. T. Suzuki (Eds.), Metallothioneins, VCH, New York 1992
  2. M. J. Stillman, Coordination Chem. Rev. 1995, 144, 461.
  3. J.F. Riordan, B. L. Valee (Eds.), Metallobiochemistry, part B- Metallothionein and Related Molecules. Methods in Enzymology, vol. 205, Academic Press, New York, 1991.
  4. K. T. Suzuki, N. Imura, M. Kimura (Eds.) Metallothionein III, Biological Roles and Medical Implications. Birkhauser Verlag Basel, Switzerland, 1993.
  5. C. Klaassen (Ed.), Metallothionein IV, Birkhauser Verlag Basel, Switzerland, 1999.
  6. M. G. Cherian, P. J. Ferguson, in Cytotoxic, Mutagenic and Carcinogenic Potential of Heavy Metals Related to Human Environment (N. D. Hadjiliadis, Ed.), 217-229, Kluwer Academic Publishers, Dordrecht, The Netherlands, 1997.
  7. R. F. Olafson, Biochem. Bioenerg. 1988, 19, 11.
  8. A. Munoz, A. R. Rodríguez, Electroanalysis 1995, 7, 674.
  9. C. Ruiz, J. Mendieta, A. R. Rodríguez, Anal. Chim. Acta 1995, 305, 285.
  10. M. Fedurco, I. Šestáková, Bioelectrochem. Bioenerg. 1996, 40, 223.
  11. C. Riutz, A. R. Rodríguez, Anal. Chim. Acta 1997, 350, 305.
  12. Ch. Harlyk, O. Nieto, G. Bordin, A. R. Rodríguez, J. Electroanal. Chem. 1998, 458, 199.
  13. Ch. Harlyk, G. Bordin, O. Nieto, A. R. Rodríguez, J. Electroanal. Chem. 1998, 446, 139.
  14. I. Šestáková, H. Vodičková, P. Mader, Electroanalysis 1998, 10, 764.
  15. O. Nieto, A. R. Rodríguez, Electroanalysis 1999, 11, 175.
  16. I. Šestáková, D. Miholová, H. Vodičková, P. Mader, Electroanalysis 1995, 7, 237-245.

4. Development of new analytical methods for trace elements

New spectrometric (AAS, ICP/MS), separation (HPLC,CZE) and electrochemical methods (differential pulse polarography, differential pulse voltammetry at mercury, glassy carbon and carbon paste electrodes, adsorptive stripping voltammetry etc.) suitable for the determination of trace elements in various matrices will be developed.