You are on page 1of 32

Trace Elements and Electrolytes, Vol. 27 - No.

3/2010 (150-180)

7th International symposium on trace

elements in human: new perspectives
October 13th - 15th 2009, Athens, Greece
2010Dustri-veriag Dr. K. Feiste

O r g a n i z e r s . Pollet a n d S. Ermidou-Pollet Athens, G r e e c e

ISSN 0946-2104

The diagnosis of trace element

deficiency in iiumans in developing countries
M. Abdulia' and S. Shukla^
'Trace Element Institute for
UNESCO, Lyon Cedex, France and
^Department of Zoology, Jiwaji
University, Gwalior, India
From a public health point of view,
it is essential to assure the general
population of all countries in the
world that the intake of all nutrients,
including trace elements, is adequate
in the average, normal daily diet. At
the same time, the ideal diets should
not contain more than the permitted
levels of toxic heavy mlais. Except
for occupational exposure, the main
pathway through which trace elements enter the human body is via the
food chain. Severe deficiency that requires immediate medical care occurs
rarely in both developed and developing countries. On the other hand, there
are clear indications that marginal deficiencies of trace elements such as
iron, iodine, zinc and selenium are
fairly common in fast growing countries in Asia, Africa and South America. According to a recent World
Health Organization (WHO) report,
nearly 2 billion people in developing
countries are affected by iron and iodine deficiency alone. The prevalence
of zinc and selenium deficiency may
be of the same magnitude, but unrecognized. Iron and iodine deficiency
can easily be detected by simple laboratory tests and clinical signs. Such
tests are not available for the detection of other essential trace elements
such as zinc. Even when the dietary
intake is restricted due to starvation
and other socio-economic conditions,
normal body functions are maintained

for a certain period of time by homeostatic mechanisms and by making use

of the body reserves. Plasma/serum
levels of trace elements may not always indicate the body status. Specific enzyme assays that may reflect
the status of certain trace elements
may not be ideal for the poor population of developing countries. Population explosion, poverty, pollution and
illiteracy are the major problems facing the developing world. Trace element problems, as a major health issue have a low priority in many poor
countries. An ideal approach to study
the long-term effect of poverty and
malnutrition that influence trace element status is to follow vulnerable
groups such as children and fertile
women in the general population for
extended periods for possible signs
and symptoms of deficiency. Another
important vulnerable group is the elderly. By 2050, a third of the population of affluent countries will be aged
over 60 years. Once these people retire, they can expect to live for another
20 years or even more. Growing rank
of senior citizens is a sign of progress
but they also bring a range of problems. Deficiency of a number of
micronutrients including trace elements in the elderly has been extensively studied in Sweden by the current authors. The intake of a number
of vitamins and trace elements are low
in the elderly in Sweden compared
with the current recommendations.
There are a number of social and economic problems associated with low
intake levels. Another practical approach of detecting the existence of
trace element deficiencies rests in
therapeutic trials. Tbe response to
iron supplementation in iron-deficiency anemia is a good illustration
showing the impact of supplemen-

tation. This presentation is aimed at

discussing practicable approaches in
detecting trace element deficiencies
at an early stage in vulnerable groups
in the general population, especially
in the fast-developing countries in Africa, Asia and South America.

Use E-iearning technology and

cybernetic methodology for
modern education in the area
of prevention of environmental
health hazard based on
sustainable development
R. Tadeusiewicz and
J.W. Dobrowolski
AGH University of Science and
Technology, Krakow, Poland
Prevention of environmental healtii
hazard as a top priority of sustainable
development requires common action
of experts and knowledge-based society. Modem e-leaming is a necessary
tool for adequate life-long learning
about innovative methods of early detection e.g. deficiency or excess of
some trace elements in the human environment, diet and body, as well as
for dissemination of innovative technologies for more effective primary
prevention. The principles of modem
distance education-including cybernetic methodology, modeling leaming, relation between knowledge
sources, leaming environment and
knowledge representation language,
notion and structure are the basic subject of this report.


Ethical sustajnability in the

research environment
L.M. Gallicchio
Rutland Institute for Ethics,
Clemson University, Clemson, SC,
Human values are inextricably
part of the successes in scientific research, yet they are not often appreciated enough and focused on. These
values constitute the ethos of the research enterprise within which you
operate your research program. Ethical sustainability, or EtS, is a way to
enhance your program to assure that it
endures over time. EtS also helps you
impact the next generation of scientific leaders by involving your team
members in defming, maintaining
and teaching EtS to others who join,
or work with, your research program.
F.O.O.D. is a tool to assist in achieving EtS. Fidelity, Openness, Ownership and Diligence are key values in a
healthy ethos. EtS and F.O.O.D. are
essential to organizational health the
same way trace elements are essential
to human health. Without them, your
research program may not die immediately, but it is more prone to "illness" and a shorter, less optimal life.
Taking the initiative to focus on EtS in
your program will contribute to the
continued longevity of the larger scientific research enterprise that has unlocked innumerable secrets to our existence.

Metaliomics: a fundamentai
approach in mechanisticaiiybased metai toxicoiogy
E. Sabbioni
Ageing Research Center (CeSI),
"G d'Annunzio" University
Foundation, Chietl, Italy
Scientific research on biometals
has generally been carried out separately in many fields such as biochemistry, nutritional science., medicine,
pharmacy, toxicology, agriculture and
environmental science, with "metals"
as common factor. However, a better
promotion of the biometals science as
interdisciplinary field to integrate the
metal-related scientific areas requires
a more general approach, in this context, metaliomics has been recently

proposed as a new frontier in the investigation of trace elements in biology [1]. Metal ions are utilized by
fundamental cellular processes. In
this context, transcription, translation
and incorporation of the adequate
metal to function are regulated by the
organism, being synthesis and metabolic function of genes (DNA, RNA)
and proteins not possible without the
aid of metal ions and metalloenzymes. Thus, the biochemistry of a cell
needs to be characterized not only in
its peculiar genetic (genome) and protein (proteome) context but also by
the distribution of all chemical forms
of the metals/metalloids (metallome)
at different levels of complexity
(whole body, organ., cell and individual biomoIecule).Thus, the purpose
of metaliomics (study of metals and
metal species and tbeir interactions,
transformations and functions in biological systems) is to extend and
broaden the bioinorganic chemistry
as a "systematic scientific field" and
an interdisciplinary science complementary and in symbiosis with other
"omics" such as genomics, proteomics and metabolomics. From the experimental point of view, metaliomics
involves the determination of a metal
in whole cells, its distribution over
different cell compartments, its association with particular pools (macromolecules, small molecules, free
ions) and the binding to a specific
class of biomolecules. These versatile
measurements require the availability
of advanced and sophisticated analytical tools to reach an adequate specificity of the analytical response with
regard to the identification of a particular metal species, and enough sensitivity to allow the ultratrace detection
and quantitation of covalently bound
metals in microsamples of proteins
and metal-protein complexes. As
such, metaliomics, referred to cellular
biochemistry, can be considered a
subset of speciation analysis (distribution of an element among defined
chemical species in a system). The
objectives of this work are to present
examples of an experimental in vitro
mechanistically-based dynamic metaliomics research. In particular, the
aims are (i) to highlight the role of nuclear and radio analytical techniques
(neutron activation analysis and use
of radiotracers with high specific radioactivity) in combination with advanced spectochemical (GF-AAS,

HPLC-ICPMS and EPR techniques)

and bioanalytical techniques (gel filtration, differential centrifugation, ion
exchange chromatography, ultrafiltration) that allow overcoming of
great analytical difficulties related to
the study of metallomes in microsamples of cell culture media, intact
cells, subcellularfractionsand isolated
biomolecules (ii) to show typical results concerning: (a) cytotoxicity and
carcinogenic potential induced in
mouse fibroblasts Balb/3T3 cell line
by different chemical species Cd, Cr,
Pt and V (b) neurotoxic effects induced by inorganic As in rat brain reaggregates and their individual cell
types as well as by Mn species in rat
pheocromocytoma PC12 cell line in
relation to their uptake, intracellular
distribution, and binding to biomolecules. The examples reported show
tbe complexity of metaliomics research and its strong multidisciplinary


Haraguchi H. Metaliomics as inlegrated

biomdical science, J Anal At Spectrom.
2004, 19:5-14.

The forensic pharmacoiogy of

inorganic poisons used in the
19th and 20th centuries
N.J. Birch'-^
'Academic Consultancy Services
Ltd, Codsall, Staffordshire and
-University of Wolverhampton,
Inorganic substances have been
used as poisons since early in man's
history. Poisons have always been a
means of injuring or killing an opponent without apparent contact or connection between the assailant and his
(or very often her) victim. Physical
strength is not required and the attack
can be planned according to opportunity, which may be remote in time or
place. Poisoning is, by definition, premeditated. The pattern of criminal
poisoning has changed over the last
two centuries largely as a consequence of the developments in analytical techniques, systematic forensic
investigation, knowledge and recognition of natural and unnatural causes
of death and increased regulation of
the supply of poisonous materials.

7th International symposium on trace elements in human: new perspectives, October 13th - 15th 2009, Athens, Greece

This largely mirrors the progress in

conventional medicine itself where
increasing regulation ofthe practitioners and their materials and techniques has led to an immeasurable reduction in adverse effects and deaths
caused by the treatment rather than
the disease itself At the commencement ofthe 19th Century poisoning
was commonplace using cheap, easily available materials such as arsenic,
sold widely for the killing of vermin,
and detection was unlikely because
systematic investigation of disease
and ofthe causes of death had hardly
begun. A decedent was as likely to be
wrongly diagnosed as poisoned.,
when dying of natural causes, as the
converse. The chances of escaping
justice were significant since many
deaths were ascribed to common infectious diseases which were then untreatable. However, those punished
were equally likely to be innocent and
wrongly accused of what was, in fact,
a natural death. Despite the major
changes in the detection and prevention of poisonings there are still in
present times, very occasionally, notorious cases which highlight specific
weaknesses in our present justice and
medical systems, I shall discuss some
interesting cases past and present to
exemplify how regulatory and investigational change has tried to keep
up with novel poisons and how
poisoners have adapted their methods

Trace elements and alternative

medicine - an awareness
S.N. Rehman
The University of Texas,
Brownsville, TX, USA
Long before the creation of the
modem medical universities in the
West, the ancient civilizations in
China, India, Southern Europe, Africa, and the Central Americas have
been preventing and treating disease
with natural foods, herbs, roots,
shrubs, and methods other than surgery. This paper compares some of
these ancient practices (now known
as alternative medicine) with our
"standard" Western medical practices. Not only has Western medicine
ignored the ancient medical practices
and wisdom but has also labeled it as

"ahemative medicine" thus giving it

the second class citizen status. Primarily, this paper attempts to accomplish three goals: one, it describes the
importance of trace elements in human health; two, it offers a short description of alternative medicine in
general and herbal medicine in particular; and three, it presents the findings
of a pilot study about awareness of alternative medicine in the United
States of America. It is the hope of this
writer that a research agenda would
emerge for developing promotional
campaigns to increase awareness of
trace elements and benefits, risks, and
availability of herbal alternatives.


Mineralimbalancen group in Berlin a

significantly increased risk for stroke
(16 %) was observed. In addition
myocardial infarction (5 %) was less.
A substitution with magnesium improved clinical symptmes in those
magnesium deficient patients. In a recent study of our group the benefit of
sartans and the new renin inhibitor
aliskiren was described while showing no negative effect on magnesium
metabolism in hypertensive patients.

Lithium - still interesting after

all these years
V.S. Gallicchio

Magnesium metabolism and

cardiovascular diseases
K. Kisters', D. Liebscher' and
M. Hausberg'
'Medizinische Klinik I, St. Anna
Hospital, Heme, -SHO
Mineralimbalanzen, Berlin, and
^Nphrologie, Stadt. Kliniken,
Karlsruhe, Germany
An overview of magnesium metabolism and the role of magnesium in
development of primary hypertension
is given. Magnesium is an essential
electrolyte in living organisms, which
has to be substituted in a sufficient
amount. The clinical importance of
magnesium overload or magnesium
intoxication is seldom. However, magnesium deficiency is of special importance in humans, despite normal magnesium supplementation. The primary
effect of a magnesium deficiency results in a reduction of several enzyme
activities in metabolism and energy
production. The reduced energy production can result in a disturbed membrane function, calcium magnesium
antagonism and cell dysfunction.
Thereby consequences may result in
an organ dysfunction and an altered
answer to extemal and internal stress.
The reduced energy status is responsible for the recovery of unhealthy individuals, e. g. cardiac arrhythmias, primary hypertension, pre-eclampsia,
cramps, allergic reactions etc. The
special importance of an oral or intravenous supplementation of magnesium has often been discribed in a variety of diseases. In an epidemiologic
investigation of 38 patients with magnesium deficiency of the SHO-

Institute for Environmental

Toxicology, Department of
Biological Sciences, Clemson
University, Pendleton, SC, USA
Lithium (Li) salts have been
widely used in psychiatry as mood
stabilizing agents for 60 years. Li is
found in variable amounts in foods,
especially grains, vegetables, and in
some areas, the drinking water provides a significant source of the element. Therefore, dietary intake in humans depends on location, type of
foods consumed, and fiuid intake.
Traces of Li have been detected in human organs and tissues, leading speculation that the element was responsible for specific functions in the human body. It was not until the 20th
century with studies performed in the
197O's and I990's, primarily in rats
and goats, that maintained on Li-deficient those diets demonstrated higher
mortality, altered reproductive and
behavioral abnormalities. Such deficiencies have not been detected in humans; however, studies peribmied on
populations living in areas with low
Li levels in water supplies have been
associated with higher rates of suicides, homicides, and the arrest rate
for drug abuse and other crimes. Li
appears to play a significant role in
early fetal development as evidenced
by high Li levels during the early gestational period. Biochemically, the
mechanism of Li action involves
multifactor and interconnected pathways with enzymes, honnones, vitamins, and growth and transforming
factors. This body of evidence now
appears sufficient to label Li as an essential element with the recommended RDA for a 70 kg adult of



1,000 mg/day. Of extreme importance

for the future is the growing body of
evidence indicating Li can be used effective for the treatment of acute brain
injuries, e.g., ischemia and chronic
ncurodegenerative diseases such as
Alzheimer's disease, Parkinson's disease, Tauopathies, and Huntington's
disease. This conclusion is based
upon evidence showing Li is important in neurogenesis as well as protecting neurons from neurotoxicity.
Li influences stem cells, both neuronal and marrow derived, thus additional therapeutic implications for
this element in clinical medicine may
be to treat disorders associated with
the faulty production of blood and
nerve cells or as a tool to enhance
blood stem cell mobilization for

Selenium and human cancer current status and future

W. Wasowicz
The Nofer Institute of Occupational
Medicine, Department of Toxicology and Carcinogenesis, Lodz,
There is a growing interest in the
biological role of selenium (Se) with
respect to both protection of human
health and prevention of diseases.
Food is a major source of Se and researchers bave a vested interest in Se
status in various populations not only
because of its deficiency or toxicity
symptoms, but also in view of its potential beneficial effect contributing,
among others, to cancer prevention.
Epidemiological studies, including
retrospective., prospective and also intervention, sbow that a low Se level
may increase the risk of certain cancers. However, it should be noted that
there is also a relatively large number
of studies, in which no effect of Se on
cancer has been observed. In the light
of recent studies, it may be assumed
that not only low but also high Se status may be associated with an elevated cancer risk. Thus, evidence for
thechemopreventiveroleof Se, based
on the current epidemiological data,
seems to be conflicting. In order to
find an accurate explanation of this
divergence, researchers have begun
to study the role of Se in the development of cancer at the molecular level.

As selenium is present in the human

organism mainly in the selenoproteins (in the form of the selenocysteine - 21st amino acid encoded
by DNA), the studies on genetic polymorphism of these proteins in terms
of cancer risk assessment are particularly attractive to be conducted. It has
been found that certain genetic variants of the selenoproteins' encoding
genes (e.g., GP x i, GP x 4, SeP) may
modify cancer risk. However, the results of these studies also remain confiicting, and no general conclusions
can be reached on the diet related
carcinogenesis and antioxidant status
(including intervention study) on cancer risk. To sum up, the majority of
studies on the relationship between
selenium and cancer, focus either on
the association between Se status and
cancer risk or on the association between genetic polymorphism of
selenoproteins' genes and cancer risk.
Combining both types of data (concerning both dietary and genetic factors) would be more informative and
valuable in the assessment of cancer
risk development, which was indicated in recent studies. On the basis of
recent findings, I would like to conclude that studies on the relationship
between diet and cancer should focus
on the interactions between dietary
and genetic factors rather than on the
study of each factor separately. Data
from such studies would be especially
interesting in view of the intervention
trial planning. Prior to supplementation, DNA genotyping should be
first performed in order to select individuals with certain genetic background. This would allow to avoid (at
least to some extent) the study bias associated with genetic variation and to
identify individuals who, due to the
specific gene and nutrient interaction,
are more or less susceptible to cancer.

This study was supported by
ECNIS Network of Excellence (Contract No. 513943/UE).

Trace elements in health and

diseases: cancer/inflammation
T. Westermarck', T. Tallberg-,
V.M. Abdulla\ M. Crohns' and
F. Atroshi"
'Rinnekoti Research Centre, Espoo,
'The Helsinki Institute for
Bio-Immunotherapy, Helsinki,
Finland, 'UNESCO, Lyon, France,
and -^Pharmacology and Toxicology
(ELTDK), University of Helsinki,
Helsinki, Finland
Inflammation and cancer. New
cancer incidence remains high worldwide. Oncologists are paying more attention to the role of inflammation in
chronic diseases. The possibility that
mutation and inflammation are mutually reinforcing processes that, left
unchecked, can transform normal
cells into potentially deadly tumors
through macrophages and other inflammatory cells with oxygen free
radicals. Inflammation may develop
into a systemic response characterized by changes that include fluctuations in the levels of plasma proteins,
trace metal, hormones, and electrolytes. A number of pathologic, environmental, psychological, and nutritional factors have been associated
with chemotherapy-related fatigue. In
people with cancer, three major
mechanisms may be involved: alteration in the body's ability to process
nutrients efficiently, increase in the
body's energy requirements, and decrease in intake of energy sources.
Tbe trace elements play their most important role as cofactors in enzymes.
It can play a role in tumor suppression
by stimulating an antitumor immune
response, but more often it appears to
stimulate tumor development. Cancer patients should avoid ingesting
certain natural dietary components. Certain amino acids and essential metal ions may act as growth factors for primitive malignant cells.
Cancer patients should therefore
avoid ingesting everything bought
from the health food shop since that
could introduce specific cancer
growth factors. The metabolic deficiency to be corrected in cancer is not
caused by starvation but actually due
to the lack of a few very specific dietary components forming the regulatory complexes in the patients' serum
(Figure). Recommended duration
for dietary measures and the use of
vaecines are long. The chronic meta-

7th Internationai symposium on trace elements in human: new perspectives, October 13th - 15th 2009, Athens, Greece
chmlcBl groups of amIno add
wed as dietary supplment:
Ala, It, Lflu. Val I Arg, His, Lye I
TTir, Sir I Glu, Atp) Met, Cye I Try. Phe I Pro

Practicable Radioisotopes
N.V. Phlllps-Duphar, compiled by J.JJ^men







Tl V Cr Mn Fe,Co






Figure 1. (Westermarck et ai.)

bolic deficiency causing cancer

seems to exacerbate as a function of
time, consequently requiring slightly
increased dosages ofthe dietary supplementation, for every decade in
therapy. This may then sustain the patients' health. Efforts to compensate
the etiologic metabolic deficiency in
cancer patients' should preferably
start before surgery, since it can prevent spread and implantation mtastases. It also seems to shorten the convalescence. The administration of a
small surplus of certain essential natural food components can also compensate genetic weaknesses, as has
been shown in cases suffering from
e.g. HNPCC, or BRCA. The possibility to compensate an inherited weakness by biological means should actively be taken into account, and acted
on. Active biological compensation
trials, instead of attempts to correct
inherited genetic ailments, deserve
more clinical studies. If gene therapies ever become successful in adults
it would be too expensive of a treatment modality for world populations,
presently suffering from ever increasing cancer incidences [I]. Explanation for the complete regression of tumors following biotherapy. No consistent tumor-specific immunological reaction could be
found which could explain the CR observed. Inconclusive evidence was
obtained from extensive immunological screening for circulating and/or
cell-bound immunity, cytolytic antibodies, effector T-cells, macrophage

inhibitor factor, natural killer cells,

killer cell indexes, tumor cell lysis,
apoptosis, phospholipase or complement activation, enhanced interferon
production or chalone reactions etc.
During the healing reaction obtained,
it was found that organ-specific mitochondria were activated [1]. Certain
test-animals have not been able to
sustain health if fed only specific gene
manipulated food-items possibly
since mitochondria do not recognize
them as belonging to their phylogenetic repertoire. Radiation as a means
to preserve food may also be deleterious since it kills the mitochondria in
our diet - the grass-roots on which we
thrive? A low temperature inactivates
also regulatory mitochondria. In former times, blood donors were transferring warm blood to the recipient,
and surgeons observed then that the
cancer growth stopped for a while. As
we are wann-blooded the mitochondrial function also seems to require a body temperature to act properly?


Complementary and alternative

medicine (CAM) used in terminal ill patients
J. Bntzel'^ H. BntzeF, O. Micke^^K. Kisters^^F. Bruns-^,
M.GIatzel''\ R.Mcke',
K.G Schnekaes** and U. Schfer'-^
'Department of Otolaryngology,
Head Neck Surgery,
^Department of Palliative Medicine,
Sdharz-Krankenhaus Nordhausen
gGmbH, Nordhausen, 'Department
of Radiotherapy, Franziskus-Hospital, Bielefeld, ""Department of Internal Medicine I, St. Anna-Hospital,
Heme, ^Department of Radiotherapy, Special Oncology, Medical
School Hannover, Hannover,
""Department of Radiotherapy,
Zentralklinikum Suhl gGmbH, Suhl,
^Department of Radiotherapy,
Klinikum Lemgo-Lippe, Lemgo,
and "AKTE - The German Working
Group "Trace Elements and
Electrolytes in Oncology"

Objective: The use of complementary and alternative methods is common in oncology, but nothing is known
about the role of CAM in the last days
of a patient's life. Material and Methodx Between 1-2007 and 10-2008 the
palliative care team treated 85 patients (50 male, 35 female, median
age 62 years) on out-door basis. All
families were asked to complete a
structured questionnaire regarding
the use of CAM during the last period
of a patient's life. Patients suffered
from cancer in 83/85 patients (98%),
two patients had neurologic disorders. Results: Only 20 families (31 %)
have not used any CAM. Spiritual
support (praying, hospice services)
was for asked by 60/85 families
(71 %). 25/85 families (29%) reported
drug use. Favorite methods were vitamins (n = 11, 13%), trace elements
(n - 9, 11%), and mistletoe (n - 8,
9%). 30 patients (35%) were treated
by physiotherapy (28 oncology patients, 2 neurology patients). LogoReferences
[ 1 ] Tallberg Th. Westermarck T, Atroshi F. A pedy and ergotherapy were used in 10
patients (12%). 12 families (14%) repossibility to prevent recurrent melaported aroma-therapy in patient's care.
noma.renal. breast and prostate cancer J.
Main information source was the GP
Trends in Biomedicine. 2008; 3: 44-54.
(house physician) (45/85, 53%)), followed by the palliative care team
(30/85, 35%) and pharmacies (27/85,
32%). Internet, journals or self-service information were used by only
25/85 families (29%). The effectivity
of the categorized methods was esti-


mated between I and 5 (very good worthy) by the patients or their families. The following ranking was registered: spiritual support 2.4; physical
therapy 2.7, Iogopedy/ergotherapy
3.1, drugs 3.3. 65/85 families (76%)
were satisfied with the information
received from the health care practitioners. Conclusion: Complementary
methods are often practiced during
the last days of a patient. Families and
friends are looking for valid information and need help by the professionals. The different methods have to be
evaluated regarding effectivity as
well as acceptance by the patients.

Trace elements selenium and

zinc as tumour markers in patients witii advanced head and
neck cancer
J.Bntzel'^U. Knolle',
A. Garayev', R.
U. Schfer^\K.
K.G Schnekaes^ R. Hunger**,
F. Bruns''', M. GlatzeH" and
O. Micke"' '
'Department of Otolaryngology,
Head Neck Surgery, SdharzKrankenhaus Nordhausen gGmbH,
Nordhausen, -Department of Radiotherapy, Klinikum Lippe-Lemgo,
Lemgo, ^Department of Intemal
Medicine I, St Anna Hospital,
Heme, ^AKTE - The German
Working Group "Trace Elements
and Electrolytes in Oncology",
^Department of Radiotherapy,
Special Oncology, Medical School
Hannover, Hannover, ''Department
of Radiotherapy, Zentralklinikum
Suhl gGmbH, Suhl, and ^Department of Radiotherapy, FranziskusHospital, Bielefeld, Germany
During the last years we have collected data showing a relation between the status of the disease and the
serum concentrations of different
trace elements. Are they able to monitor the disease as tumor markers? In a
first study 100 untreated head/neck
cancer patients were characterized by
decreased levels of selenium, zinc and
iron. Only copper was increased.
During the radiotherapy no changes
of serum concentrations were seen. A
further analysis had shown the differences between resectable and nonresectable tumors. The larger the primary tumors were, the more de-

creased the concentration of selenium
in the serum of our patients was.
These fmdings are supported by new
data of NMR-tumor volumetry and
trace elements concentrations in 21
patients. The relation between serum-selenium and tumor volume is
stronger than between classical tumor
markers (SCC; CEA, CYFRA 21-1)
and the tumor volume. A last investigation has suggested that a destroyed
homeostasis of trace elements is a
sign of pre-terminal ill patient. When
the destroyed homeostasis is observed, the patient has a window of
4 - 6 weeks of remaining life time.
Conclusions: Trace element status offers a lot of information about the
course of the disease and the individual. But environmental factors have
more impact on this trace element status than on other classical tumor

Rare-Earth metal compiexes

with 4-hydroxycoumarins are
more active against HIV-1 in
cell culture
P. Genova-Kalou', S. Raleva^,
A. Hinkov\ 1. ManoloV,
S. Gurkova^ and R. Argirova^
'National Centre of Infectious and
Parasitic Diseases, Department of
Virology, Laboratory of Cell cultures, -National Centre of Infectious
and Parasitic Diseases, Department
of Virology, Laboratory of
Retroviruses, ^Sofia University
"St. Kliment Ohridski", Faculty of
Biology, Laboratory of Virology
and, 'Sofia Medical University,
Faculty of Pharmacy, Department of
Chemistry, Sofia, Bulgaria
Introduction: Coumarine derivatives have a broad spectrum of biological activities, including anti-HIV effect (anti-integrase-lN) recently reported. Their potential to form complexes, esp. with some lanthanides
showed antitumor and other biological activities. Seventeen bis-(4-hydroxycoumarin) (4-hc) derivatives
were synthesized in Bulgaria and later
used as ligands. First anti-HIV assays
were performed in cell culture in
non-toxic concentrations showing
that three out of seventeen 4-hc had
anti-HIV activity (IM-7, IM-8 and
IM-10)(IM-7, IM-8 and lM-10). This
prompted us to prepare complexes

with - Ce(III), lanthanum - La(III)

and neodymium - Nd(III) and to evaluate the cytotoxicity of the active derivatives and their complexes on
MT-4 cells and to study for anti-HIV
activity in cell cultures. Materials and
methods: Cell toxicity (CC50) and
maximal non-toxic concentrations
(MNCs) were performed in MT-4
cells on 72h by MTT-test. The MNC
and CC50 for each compound were
calculated from dose-response curves.
Reverse transcriptase (RT) activity
and RT processing (PCR) were studied to see the impact on early stages of
HIV-1 replication. Results: The less
cytotoxic compound, according to
^4NC, was IM-8, whose MNC values
were higher than those of IM-7 and
IM-10. The less cytotoxic compound,
according to CC50 values, was IM-8
for all derivatives. Six out of seventeen complexes showed higher HIV-1
inhibition compared to that of the ligands. Ce(lll) complexes were more
prone to inhibit the HIV-activity in
cell culture than La(I[I) and Nd(III)
complexes. Conclusions: Rare-earth
metal complexes are readily formed
with 4-hc. Complexes with Ce(III)
show higher anti-HIV activity than
those with La(llf) and Nd(III). Further studies are in progress to detect
the target and mechanism of action.

Trace element disturbances in

platelets of juvenile and infantiie NCL patients
T. Westermarck', E. Johansson' and
F. Atroshi^
'Rinnekoti Research Center, Espoo,
and -Dep. Pharmacology and
Toxicology, ELTDK, University of
Helsinki, Finland
Depletion of ATP/PC stores is an
essential trigger for all subsequent
cytotoxic events in focal cerebral
ischemia formed e.g. during seizures.
Maintenance of ionic gradients requires significant energy; with the reduction in ATP, there is a gradual increase in intracellular Na+ and Ca-H-,
resulting in cell depolarization, and
release of glutamate, which secondarily stimulates the AMPA and
NMMDA receptors, opening ionic
channels allowing a rapid influx of
Ca++ into the cell. This increase stimulates the initiation of proteolysis,
lipolysis, and DNA degeneration, that

7th International symposium on trace elements in human: new perspectives, October 13th - 15th 2009, Athens, Greece

if continued, produces cell death and

neurological injury associated with
cerebral ischemia, and probably preaging degenerative changes seen in
Batten's disease (NCL). Disorders in
which free radicals and antioxidants
are implicated include Batten s disease. Recent evidence has shown the
importance of diet in delaying the aging process and in protecting against
several degenerative and chronic diseases. Recent cortieosteroid trial
(Prednison 40 mg x 1 ) ofNCL patients
has increased the need of checking the
trace element levels of the treated
NCL patients, when corticosteroids
may result in increased need of
supplementation of calcium, magnesium and zinc; and speed up the cognitive deterioration. We have examined trace element levels in platelets
of infantile and juvenile NCL patients. Metal ions from platelets were
analyzed by micro-PIXE; metal ions
from erythrocytes by ICP-MS. The
concentration of Ca was significantly
increased, and Zn significantly decreased in the throbomcytes in juvenile, but not in infantile NCLpatients.

Therapeutic metabolic and

kinetic aspects of ianthanum
carbonate: a new phosphate
binder in chronic renal failure
PC. D'Haese, A.J. Bervoets,
M.E. De Broe and GJ. Behets
Laboratory of Pathophysiology,
Faculties of Medicine and Biomedical and Pharmaceutical and
Veterinary Sciences, University of
Antwerp, Antwerp, Belgium
In the search for safe alternatives
for calcium- and aluminum-based
agents lanthanum carbonate has recently been introduced as a new phosphate binder in chronic renal failure
patients. Clinical studies showed the
compound to be highly efficient for
phosphate binding. In view ofthe past
experience with aluminum however,
some concerns have been raised as to
whether lanthanum treatment might
go along with some toxic side effects,
particularly at the level of the bone.
Clinical as well as experimental studies indicated that although there is
some minor deposition ofthe element
in bone, long-term treatment is not accompanied by any mineralization de-

fect or the development of a low bone

turnover disease. Moreover, in contrast to aluminum, ultrastructural localization studies revealed lanthanum
deposition not to be associated with
any type of renal bone disease. As lanthanum is mainly excreted by tbe
liver, patients with chronic renal failure are not at an increased risk for accumulation of the element as compared to subjects with nonnal renal
function. After oral treatment however, we and others noticed 2 - 3 fold
higher lanthanum levels in the livers
of rats with chronic renal failure compared to rats with normal renal function. Studies dealing with the kinetics
and tissue distribution, absorption,
and subcellular localization of lanthanum in the liver using transmission
electron microscopy, electron energy
loss spectrometry, and X-ray fiuorescence showed that in the liver lanthanum was located in lysosomes and in
the biliary canal, but not in any other
cellular organelles. This suggests that
lanthanum is transported and eliminated by the liver via a transcellular,
endosomal-lysosomal-biliary canicular transport route. Feeding rats with
chronic renal failure orally with Ianthanum resulted in a doubling ofthe
liver levels compared to rats with normal renal fiinction, but the serum levels were similar in both animal
groups. These levels plateaued after 6
weeks at a concentration below 3 jig/g
in both groups. When lanthanum was
administered intravenously, thereby
bypassing the gastrointestinal tractportal vein pathway, no difference in
liver levels was found between rats
with and without renal failure. This
suggests that there is an increased
gastrointestinal permeability or absorption of oral lanthanum in uremia.
Lanthanum levels in the brain and
heart fluctuated near its detection
limit with long-term treatment (20
weeks) having no effect on organ
weight, liver enzyme activities, or
liver histology. We suggest that the kinetics of lanthanum in the liver is
consistent with a transcellular transport pathway, with higher levels in the
liver of uremic rats due to higher
intestinal absorption.


Se, Pb and Cd ieveis in renal

failure patients in China
B.Chen'% L.V.Lamberts',
G.J. Behets', T.-t. Zhao^ L. Gang-,
X.-h. Hou'% G-j. Guan^ and
PC. D'Haese'
'Laboratory of Pathophysiology,
Faculties of Medicine and Biomedical and Pharmaceutical and
Veterinary Sciences, University of
Antwerp, Antwerp, Belgium,
-Department of Nephrology,
The Second Affiliated Hospital,
Shandong University, and
'Department of Respiratory
Medicine, Shandong Provincial
Chest Hospital Jinan, P. R. China
Whole blood and serum samples
of Chinese stable chronic renal failure
(CRF) patients (n ^ 81 ), hemodialysis
patients (n = 135), post-transplant patients (n = 60) and subjects with normal renal function (NRF) (n = 42)
were collected, as well as water and
dialysate samples from 5 dialysis centers. The concentration of selenium
(Se), lead (Pb) and cadmium (Cd) was
measured by atomic absorption spectrometry. The mean serum Se levis in
patients with different degrees of renal failure were significantly lower
than those of subjects with NRF (p <
0.01). Pb levels were not increased in
renal failure patients while the Cd levels in patients with various degrees of
renal failure were higher than in subjects with NRF (p < 0.05). After correcting the result of Pb and Cd for
hematoerit (Hct) however, Pb levels
of dialysis patients were also increased. In the dialysis population under study, blood Pb and Cd levels
were closely related to the time on dialysis, while contamination ofthe final dialysate may also contribute to
the increased blood Cd and to a less
extent Pb levels. Correction for Hct
may be recommended to accurately
compare blood Pb and Cd levels in dialysis patients and CRF patients with
varying degrees of anemia to those of
subjects with normal renal function.



Diabetes Type 2:
A new perspective
D. Oberleas
Department of Food and Nutrition,
Texas Tech. University, Lubbock,
There is mounting evidence that
chromium'^ is required as a cofactor
for the normal function of insulin. An
adequate supply of insulin is synthesized and circulating in most Type 2
diabetics. For insulin to function
properly it must attach to an insulin
receptor in the cell wall and form a
stable attachment for a short period of
time. However, in the absence of
disulfide cross-linking, protein:protein interactions are not stable. A
small peptide with molecular weight
of 1500 has been isolated and functions like a chaperone by complexing
chromium'* to insulin receptors and
subsequently to insulin to form a stable union, fhc stable union creates a
quantum of light and production of
HiO:. Other symptoms associated
with Type 2 diabetes are quite variable among patients and are a result of
the variety of sub-clinical deficiencies described previously. Zinc is involved in every normal cell division
in the body. Zinc is also important in
the maintenance of normal thymus
morphology and function. Thus, secondary infections have an increased
prevalence and extended infectivity
that may result in physiological crises. Copper deficiency symptoms are
accentuated and may result in increased hemorrhagic strokes, aortic
aneurisms and cardiac aneurisms.
Many diabetics suffer from leg
cramps and have cholesterol quality
problems that may be effectively
treated with magnesium supplementation. Nickel, boron and other ultra-trace elements may also have minor contributions towards normal
physiology associated with Diabetes
II. It is not clear why these symptomologies may be drastically accentuated by the elevation of circulating
glucose whose major effect is
increased osmotic pressure.

Chronic diabetes mellitus and

level of trace elements in
A. Skesters', A. Silova',
N. Rusakova' and N. Savickiene'Laboratory of Biochemistry of the
Riga Stradins University, Riga, and
-Department of Pharmaceutical
chemistry and Pharmacognosy of
Kaunas Universitate of Medicine,
Kaunas, Lithuania
Diabetes Mellitus (DM) is a serious
and costly disease which is becoming
increasingly common, especially in
developing countries and disadvantaged minorities. Recently compiled
data of WHO shows that approximately 246 million people have diabetes worldwide, and that this number
may increase to 380 million by the year
2025. In Latvia on 1st January 2007
53,997 DM patients have been
reorded (about 2.57% of the population), however the opinions of the International Association of Diabetes,
is that in Latvia 9.9% (207,900) of the
population can be patients. DM is a
chronic disease that occurs when the
pancreas does not produce enough insulin (Type 1), or alternatively, when
the body cannot effectively use the insulin it produces (Type 2). In diabetes,
an altered oxidative metabolism is a
consequence either of the chronic
exposure to hyperglycemia of or of the
absolute or relative insulin deficit; insulin regulates several reactions involved in oxido-reductive metabolism, besides, abnormalities in the regulation of peroxide, transition metals
and trace elements metabolisma re
postulate to result in establishment of
the disease as well as its longer term
complications. DM associated with
oxidative reactions, particularly those
which are catalyzed by decompartmentalized transitions metals, but their
causative significance is diabetic tissue damage remains to be established.
On the other hand, trace elements,
such Se, Zn, Mg, Cr, Mn ect., are essential constituents of a number of enzymes, some of which have antioxidant functions. In the present study
we have detected levels ofsome trace
elements using fluoremetric method
for Se and Randox laboratories Ltd.
(UK) kits for Zn (ZN 2341) and Mg
(MG 3880) in blood of patients with
chronic DM before supplementation
with natural antioxidants. Our results
showed no considerably diferences

from normal value of Se, Zn and Mg

content in blood. Thus it can be concluded that in chronic DM patients
during the stage when the first neiro-,
retino-, and nepbropathy characteristic
mark appear, content of trace elements
do not change.

Macro and trace elements in

overweight and obesity
M.G. Skalnaya
ANO Centre for Biotic Medicine,
Moscow, Russia
Interconnection of trace element
profile and metabolic disorders causing overweight individuals are reported. In this connection, the aim of
our study was to investigate association of hair elemental profile with
obesity and Type 2 diabetes In
women. 1470 women 46 - 60 years
were subjected to study. They were
divided in three groups: control (n ^
1236, BMI 18 - 25 kg/m-), obese
women with BMl > 30 kg/m- (n =
141) and diabetic women (n = 93,
BMI 18 - 24 (19%), 25 - 30 (38%)
and > 30 (43%)). Multielement hair
analysis was conducted by ICP-AES,
ICP-MS methods. Obese women
were found to have elevated hair Hg,
K, and decreased Ca, Mg, Zn and iodine as compared to control subjects.
In the diabetic group similar deviations were found: elevated hair K, Hg,
Na and decreased Ca, Mg, Zn, Co
compared to control. The only difference between obese and diabetic
women was Na level, which was significantly higher in Type 2 diabetic
patients as compared to the control
and obese ones. It is interesting to
note that hair iodine was lower only in
obese patients, but not in diabetics.
Thus, in Type 2 diabetes there are
more numerous and serious changes
in mineral metabolism as compared to
obesity impairing anabolic processes
and water-salt balance regulation; Zn
imbalance can play an important role
in pathogenesis of both obesity and
Type 2 diabetes. The obtained data
also suggests that muhielement hair
analysis is tbe useful diagnostic tool
in prevention and treatment of excessive weight gain, obesity and metabolic syndrome in women.

7th International symposium on trace elements in human: new perspectives, October 13th - 15th 2009, Athens, Greece

Trace element imbalances role

in pathogenesis of fetal
alcohol syndrome
A.V. Skalnya
ANO Centre for Biotic Medicine,
Moscow, Russia
Alcoholism and related hereditary
consequences, including fetal alcohol
syndrome, are one of the dramatic
problems in Russian public health.
The severity of alcohol related derangements are also due to malnutrition, micronutrient deficiencies and
ecological pollution. In a series of experimental investigations, the role of
Cu/Zn imbalance in the whole body
and numerous deficiencies of macro
and trace elements with simultaneous
Pb, Cd accumulation in central nervous
system were demonstrated (Skalny,
1985 - 2008). We have found the correlation between trace element imbalances and behavioral deviations in experimental animals, and protective effects of zinc in offspring of alcoholized rats due to normalization of
trace element ratios in brain structures. Preliminary data from clinical
nutritional studies, which are ongoing
as a part of the international Collaborative Initiative on Fetal Alcohol
Spectrum Disorders (CIFASD, Prof
C.L. Keen et al.) suggests a potential
role of zinc and choline in development/prevention of fetal alcohol spectrum disorders.

The author gratefully acknowledges Dr. Christina D. Chambers,
Prof Carl. L. Keen (University of
California) and Elena S. Vyatchanina
(Russian Society of Trace Elements in
Medicine) for collaboration in FASD

The content of trace elements

in aerial parts of plants of
Genus Pulmonaria
D. Kj-uglov
Novosibirsk State Medical University, Department of Pharmacognosy,
Novosibirsk, Russia
Herbal remedies produced from
the aerial parts of plants of the genus
Pulmonaria have anti-anemic activity. A similar pharmacological activity is evoked by trace elements con-

taining an herbal raw. The trace elements composition of five species of

Pulmonaria (Pulmonaria officinalis
L., P. obscura Dumort., P mollis
Wulf ex Homem., P angustifolia L.
and P. rubra Schott) was researched
by means of inductively coupled
plasma mass-spectroscopy. It was established that trace elements composition of researched plants is similar.
Biogenic trace elements (B, K, P, V,
Ca, Co, Cu, Fe, Mg, Mn, Mo, Na, Si,
Zn) have a smaller variability in contrast to 47 others trace elements. Evidently, the content of biogenic trace
elements define the homeostasis of
vegetable organism, and variability of
this content has to be smaller than the
other elements. Accumulating of the
other elements has a random character. Different parts of a plant (generative shoots and rosellate leaves) have
a different relative amount of biogenic trace elements, and different ratio of contents of manganese and iron.
Therefore, herbal remedies produced
from different parts of a plant can
have a different pharmacological activity - antianemic and expectorant

Magnesium deficiency and increased fractional magnesium

excretion in insulin-dependent
diabetes mellitus - magnesium
loading test and blood
K. Kisters', M. Schildheuer^
A. Bauer^, K. Dick-, A. Peters^,
B. Gremmler*, M. Hausberg\
M. Quang Nguyen' and P.M. Rob^
'Medical Clinic I, St. Anna-Hospital,
^Medical Clinic !, Universittsklinikum Lbeck, IMedical Clinic I,
Stadtisches Klinikum, Karlsruhe,
and ""Cardiology, Marienhospital,
Bottrop, Germany
Background: There is experimental and clinical evidence giving importance to the relationship between
magnesium (Mg) and diabetes mellitus. The American Diabetes Society
suggested that attention be given to
the diagnostic significance of a Mg
deficiency in diabetes mellitus. Because of regional differences in Mg
suppply, we studied Mg status in the
surroundings of Lbck in healthy persons as compared to diabetics. Patients
and methods: 27 healthy controls (C)


and 27 inpatients with an insulindependent diabetes mellitus (DM) and

bad metabolic control were studied.
Mg++ was measured by atomic absorption spectroscopy (Video 12 apparatus) in serum and erythrocytes; basic
24-hour urinary Mg excretion
(b-Mg-u), fractional Mg excretion
(f-Mg-u) were calculated. In addition
the Mg loading test (Mg-It) was applied. Results: In diabetics a hypomagnesaemia was observed as compared to healthy subjects both in serum (0.73 0.04 vs. 0.83 0.04
mmol/l, means SD, p < 0.05) and in
red blood celIs(1.430.16vs.l.84
0.10 mmol/l) (p < O.OI). The b-Mg-u
was 2.75 3.62 mmol/d in DM as
compared to 5.84 2.21 mmol/d in
controls (p < 0.05); f-Mg-u was 5.83
2.73 % in DM and 3.46 1.33 % in
controls. The percent retention of the
given Mg load was 24.5 18 in DM
a n d - 4 1 12 in controls (p < 0.001).
Discussion: In DM with bad metabolic
control there is convincing evidence of
a Mg-deficiency. Higher f-Mg-u in the
case of lower S-Mg in diabetes indicate an inadequate renal Mg loss
which significantly correlates with
the amount of glucosuria as one of the
relevant pathophysiologicai meachanisms.

Trace elements in
B.J. Bolann and R.J. Ulvik
Institute of Medicine, University of
Bergen, and Laboratory of Clinical
Biochemistry, Haukeland University
Hospital, Bergen, Norway
Hemochromatosis is a hereditary
disorder characterized by increased
iron uptake fiom the gut, which over
the years may result in iron accumulation in the body and iron overload.
The condition can be treated successfully by bloodletting. Bleeding of 450
ml of blood is performed every 1 - 2
weeks until the patient's iron status is
normalized. Then bloodletting 2 - 5
times a year is continued lifelong to
prevent new accumulation of iron.
Some uptake, transport and storage
mechanisms used by iron are shared
with trace elements. The increased
iron absorption, iron overload in the
tissues, and the removal of trace elements with the bloodletting, may all
afTect trace element metabolism.


However, whether changes in trace

element metabolism or distribution
participate in the pathogenesis of iron
overload tissue damage is not known.
The aim of this work is to study how
trace elements homeostasis is affected
by the bloodletting used to treat
heniochromatosis. We recruited outpatients from the Hemochromatosis
Clinic at the Department of Medicine.
After genotyping we measured iron
status and common clinical chemistry
tests, and trace element profiles were
studied before and after treatment for
iron overload. According to preliminary results, serum concentrations of
aluminium, boron, selenium, strontium and zinc decline after bloodletting. The pathophysiological consequence of this is not known.

Serum trace elements and

brain atrophy in aicohoiics
E. Gonzalez-Reimers',
C. Martin-Gonzalez',
L. G alindo-Martin',
E. Garcia-Valdecasas',
I. Hemndez-Betancor',
C. Fernandez-Rodriguez',
P. Abre-Gonzlez'
and N. Faria
'Servicio de Medicina Interna,
Hospital Universitario de Canarias,
^Dpto de Quimica Analitica, and
^Dpto. De Fisiologia, Universidad de
La Laguna, La Laguna, Tenerife,
Canary Islands, Spain
Chronic alcoholics are at risk of
developing several neurological alterations, the so-called alcoholic dementia, related to brain atrophy, and
cerebel lar atrophy, together with thiamine deficiency related Wemieke'sKorsakoff encephalopathy, being the
most frequently reported alterations.
Although pathogenesis is still obscure, cytokine-mediated neuroinflammation and oxidative damage
may play a role in brain atrophy of uncomplicated alcoholics. Therefore,
alterations in trace elements, such as
selenium and zinc deficiency, involved in antioxidative capacity, or
iron excess, frequently observed in alcoholics, acting as a pro-oxidant, may
be all involved as pathogenic factors.
In order to deepen our knowledge
about the relation between brain atrophy and trace elements and oxidative
damage in alcoholics we performed

the present study, including 61 alcoholic patients and 12 age and sexmatched controls, all of them sanitary
workers who had undergone a cranial
scan because of minor symptoms. Patients showed a more intense degree
of atrophy, differences being observed in Huckmann (t ^ 2.95), bicauda (t - 3.93) and celia index (t 3.75) and cortical atrophy (t ^ 5.45,
p < 0.001 in all cases). Statistically
significant differences between patients with cerebellar atrophy and
those without cerebellar atrophy were
observed regarding copper (854 |ig/I
181 ^ig/l vs. 1088 [ig/\ 269 ng/l,
p = 0.014), and zinc (971 ^g/1 696
Hg/1 vs. 1817 \ig/\ 1298 \ig/lp =
0.03). On the contrary, serum ferritin
was higher among patients with cerebellar atrophy (529 370 ng/ml vs.
308 209 ng/ml p - 0.044). Serum
ferritin also showed a significant correlation with bicaudate index (rho ^
0.3, p = 0.029) and Huckmann's index (rho = 0.32, p = 0.02), and serum
transferrin, an inverse one with Huckmann s index (rho ^ -0.30, p = 0.031 ).
However, selenium showed a direct
(opposite of what was expected) significant correlation with Huckmann
index (rho = 0.35, p - 0.046). No relation was observed between trace elements and age, amount of ethanol
consumed, years of consumption,
malondialdehyde (only 12 cases), or
proinfiammatory cytokines (TNF-a,
IL-6). Thus, besides ferritin, higher
among those with more intense atrophy, trace elements are only poorly related to brain atrophy, but patients
with cerebellar atrophy do have lower
serum copper and zinc levels.

Dietary bio-moduiation
schedule for treatment of
prostate cancer patients
T. Tallberg', T. Westermarck' and
F. Atroshi"
'The Helsinki Institute for
Bio-Immunotherapy Ltd., and
^Pharmacology and Toxicology,
ELTDK, University of Helsinki,
Helsinki, Finland
Prostate cancer (CaP) is a hormone-dependent disease based on
Huggins and Hobbs orchiectomy
studies in 1941 [1]. As CaP regularly
recurred, it led to bilateral adrenaiectomies [2], but in spite of this all pa-

tients died in a short time since adrenal glands actually had a central regulatory position in this endocrine disease [3]. CaP still lacks a definitive
comprehensible etiology. It is in many
cases a potentially incurable agelinked disease. Several clinical observations designate that a regulatory
function is performed by the adrenal
glands linked to a feed-back effect activated by its zona-reticularis (ZR)
cells. This positive clinical effect is
promulgated by specific dietary metabolic factors in all stages of CaP [4].
These nutritional factors have been
found to cure CaP even in cases suffering from multiple bone mtastases
[5] - to avoid hormone refractory
states (HRPC), and stop the progress
of the disease in patients on Watchful
Waiting [6]. We have used dietary
bio-modulation schedule for treatment of prostate cancer patients (Table 1). This complex dietary formula
has been developed empirically over
two decades according to the aenocarcinoma code for CaP [4, 6]. The
amino acids, trace-element ions and
CNS lipids tested were fed according
to the code while constantly monitoring the effect on the patients' hormone balance and clinical features.
Dietary bio-modulation schedule for
treatment of prostate cancer patients.
1) Oral administration of each ( 2 - 5
g/day) of respective L-amino acids;
Arg, Asp, Glu, Gly, Lys, and Ser,
eventually also His, all in connection
with meals. 2) Essential trace-element
salts prescribed orally as biologically
active ions, at dose levels of some
milligrams ( 1 - 3 mg/day); Chromium
(CrCl2.6H:O) 6 mg (- 1.17 mg Cr),
Tinn (SnCl4.5H2O) 4 mg (= 1.35 mg
Sn), Strontium (SrCb) 1 - 7mg {- 1
mg Sr), Vanadine (Na: VO.,. 4 H2O), 6
mg (- 2.5 mg V), Wolfram (Na. WO4.
2 H:0), 4 mg (- 2.3mg W). 3) Small
physiologic amounts of vitamins; A,
B, C, D, E, K, folie acid (2 mg/d) and
lycopene. To improve lymphopoiesis
and the immune-defence of patients a
diet containing prion-free neurogenic
lipids (Neurofood Ltd. Finland). All
these dietary ingredients can be mixed
together in yogurt or ice-cream, or as a
daily nutritional ration utilizing prepacked powders. Dose-levels are adjusted based on the clinical response
as measured during the therapy, and
correlated to the patients' body
weight. A good prognosis caused by
adrenal ZR feedback activation can

7th International symposium on trace elements in human; new perspectives. October 13th - 15th 2009, Athens, Greece

rapidly be accessed by standard laboratory tests, evidenced by increased

FSH, PRL, SHBG levels and the free
% of PSA, while a decrease is seen in
the levels of; DHEA, DHEAS, and total PSA-levels.


Huggins C. Hodges CV. Studies on prostate cancer: I. Effect of castration, estrogeti. and androgen injection on serum
phosphatases in metastatic carcinoma of
the prostate. Cancer Research. !')4I; I:


Huggins C. Scon WW. Bilateral adrenalectomy iti prostate cancer. Ann Surg.
\945\ 122: 1031-1041.
Tallherg T. Cancer treatment, based on
active nutritional bio-modulation, hormottal therapy and specific autologous
immunotherapy. J Aust Coll Nutr and
EnvMed. N96; 15:5-2^.
Tallbei-g T. Studies on cancer of tlie Prostate Gland, a search for Aetiological and
Prognostic Factors. J Aust Coll Nutr and
EnvMed. 2003; 22.- 11-16.
Tallherg T Regulation of cancer by therapeutic vaccination and dietary bio-modulation involving organ specific mitochondria. Int J Biotechnology. 2007; 9:
Tallherg T. Dahek M. Prostate cancer,
aetiological, therapeutic, prognostic and
prophylactic factors. Anticancer Research. 2008; 28: 3507-3508.





Bio-Immunotherapy can
prevent recurrent prostate renal cancer, and melanoma
T. Tallberg, T. Westermarck and
F. Atroshi
The Institute for Bio-Immunotherapy,
Helsinki, Finland
Tumors may spontaneously regress which implies that mammals
possess a natural intrinsic regulatory
capability to control the de-differentiation of specialized organ cells, "The
metabolic triumph of the host" (Dr.
W.M. Cole 1974). Thus this novel
paradigm is that cancer represents a
complex chronic metabolic deficiency disease which can be compensated by biological dietary means.
The aim was consequently to analyze
if the disturbed interior milieu can be
corrected /compensated and lead to a
biological cure without side-effects in
prostate cancer patients. Randomized
clinical studies with hundreds of cancer patients, over 35 years, showed
highly significant (p - 0.001) im-

provement in the survival rate by

feeding renal cancer, and melanoma
patients a specific combination of nutritional amino acids, trace-element
ions, inductional central nervous system lipids (CNS), plus supporting the
hormonal balance, and specifically in
renal cancer and melanoma by activation of the patients autologous immune-defense. The composition of
these natural etioiogical bio-modulating factors regulating our three major
forms of cancer; leukemia, adenocarcinoma, and sarcoma have primarily been outlined. Significantly better
disease-fiee intervals extending over
decades, were achieved utilizing powders containing these vital dietary supplements to correct the complex etioiogical metabolic deficiency causing
cancer. This is the "intellectual property" of this Institute. Ready-made
powders complied to treat prostate
cancer, (decreasing Gleason scores)
are available from our Institute, at a
cost of only 2 - 4 /day. In a randomized study with 127 patients suffering
from metastasized renal cancer, dietary supplements could also arrest
recurrent disease. Pertinent bio-modulating dietary components were;
L-amino acids; Ala, Arg, Asp, Lys,
plus trace-elements; Cr, Mo, Se, Sn,
V, and central nervous system CNSlipids, and physiologic doses of vitamins. Significantly improved clinical
results were also obtained with cutaneous (103 cases) and Uveal melanoma (54 patients) ingesting; Gty,
Glu, Ala, Asp, He, Lys, + Cr, Se, Sn,
V, W, and CNS-lipids. Strontium is
important to cure prostate cancer. In
bio-immunotherapy significant clinical improvement has been achieved
for dietary correction alone in renal
cancer (p ^ 0.04), and high risk (T3)
uveal melanoma (p ^ 0.001 ), but clinical results are usually further improved if the therapy is combined
with active specific immunotherapy
utilizing polymerized autologous tumor tissue. Tumor tissue should
therefore always be saved at surgery
to facilitate the preparation of individual vaccines since patients' malignant
cells contain a fingerprint of antigenic
tumor-markers. Similar good therapeutic results have also been obtained
with other forms of cancer. This healing reaction does not involve
apoptosis or lysis of tumor cells as
they regain normal healthy function,
with complete regression {CK), even


of big tumors, without a scar. Regular

immune reactions do not have such a
capacity. Actually activated regulatory organ-specific mitochondria
have been found to be involved in the
healing process. Metabolic bio-modulation can also prevent recurrent
cancer because it actively strives to
correct the etioiogical deficiency. Our
standard therapy is primarily extracting only symptoms of this metabolic
deficiency disease. It is as if only the
loose teeth of the scurvy patient were
removed instead of giving him vitamin-C. Malignant transformation
caused by genetic weakness (e.g.
HNPCC) is out of reach for gene therapy, since it involves aberrations in
several genes, but malignant transformation can be prevented by dietary
supplementation administering small
amounts of pertinent essential metabolic components aimed to regain the
physiologic internal milieu in the patients' body. Healthy persons attain
this balance from their normal diet.
All bio-modulating components involved are natural substances, and
thus ethical, inexpensive, easy to administer and have a long shelf life.
Bio-immunotherapy entails no side
effects, but also harbor prophylactic

Cancer is regulated by organspecific mitochondria via

genomics, proteomics and
T. Tallberg', T Westermarck' and
F. Atroshi'
'The Helsinki Institute for
Bio-Immunotherapy, Helsinki,
-Rinnekoti Research Centre, Espoo,
and 'Pharmacology and Toxicology
(ELTDK), University of Helsinki,
Mammalian cancer control is dependent on a complex metabolic physiologic bodily balance. Normalization of malignant gene transcription
in an organ requires dietary correction
of the etiologic long-standing metabolic deficiency involving six or more
inter-linked natural factors aided by
hormonal equilibrium, enhanced by
specific autologous immunotherapy.
In bio-immunotherapy this therapeutic bio-modulation is aimed to simulate specific leukemia, adenocarcinoma or sarcoma regulatory codes, lead-



ing to a cancer cure by forcing tumor

cells back into healthy gene transcription, without apoptosis. Mitochondria
are thus transformed and become
electron-dense as their cristae and
matrix gather metal-enzymes (Cr, Fe,
Ti, Zn) and appear in the vicinity of
the intact tumor cell nucleus. This
inductional signal is transferred to
chromosomes by strontium thru the
nuclear membrane. This physiologic
mammalian healing in hundreds of
patients resembles mysterious spontaneous cures. Immunobiology forms a
vital paradigm shift over prevailing
cancer therapies; "to kill cancer cells".
Mammalian genes (22 - 60.000) are
too few to explain the complex,
time-linked physiological expression
leading to health and senescence. The
human chromosomal nucleotide sequence analogy with monkeys and
rats (99%; 96%) is surprising, suggesting that chromosomes were created by mitochondria during eons of
their phylogenetic toil, representing a
"memory of evolution" rather than
the "secret of life". Mitochondrial action in cancer seems primarily to be
gene-regulatory and reparative, plus
its energy production for the nucleus.
They are also involved in digestive
and decay processes, with programmed cell death and ending in senescence. Organ-specific mitochondria
act as one operator in the supercomputer every cell symbolizes, where
chromosomal DNA forms the memory unit. The billions of nucleotide
chemical letters between genes are
not "junk-sequences", but they pinpoint species' specific genes during
embryogenesis. Proteomies shapes a
hundred-thousand proteins conformed
into active quaternary molecules. The
importance of the lipidome, with its zillions of "functional lipids" in our central nervous system (CNS) linked to
gestational embryonic induction, is
usually overlooked. Inductional Control and Lipidomics also functions in
the adult body but less conspicuously.
Blood-brain barrier lesions with CNSlipid loss curtail axonal cell-control in
that enervated area, espousing malignant transformation. Ingested CNSlipids stimulate lymphopoiesis, and can
alleviate diverse neurogenic ailments;
stress and pain ^ the warning signal
that CNS cannot cope with demands
from its crucial inductional signalsystem for a steady state healthy function etc. A marked depletion of CNS

lipids in affecting three specific segmentai areas of the spinal cord is possibly also linked to different forms of
clinical atherosclerosis. Mitochondria! regulation is related to lipidomics, genomics and proteomies, and
it may lead to the future use of
schooled organ-specific mitochondria as genuine cancer remedies.

Prostate cancer depends on

adrenal gland functions and
mitochondrial regulation. A
study over three decades,
analyzing etiological,
therapeutic and prognostic
T. Tallberg', T. Westermarck^ and
Faik Atroshi^
'The Institute for
Bio-Immunotherapy, Helsinki,
^Rinnekoti Research Centre, Espoo,
and 'Pharmacology and Toxicology,
University of Helsinki, Helsinki,
Epidemiology studies indicate that
dietary habits have a pronounced effect on the incidence of prostate cancer (CaP). Latent disease, which in
autopsies is found in 80% of male patients over 80 years, suggests that
metabolic factors are involved. The
aim for this study, over 30 years, was
therefore to search for metabolic
components which could reverse, and
arrest the progress of CaP. Methods:
To facilitate the dietary treatment and
improve compliance of ready-made
powders containing all pertinent metabolic factors procured and fed to patients. These powders contain specific amino acids, trace element ions
and central nervous system lipids. In
search for etiological factors 70 different cases of CaP were followed,
with informed consent, for over 10
years, recounting their clinical and
laboratory responses to this biological treatment modality. As only physiologic natural non-toxic and inexpensive metabolic components were utilized, no side-effects were encountered, but prognostic clinical features
were found. Clinical results: Following this biological treatment, transformed organ-specific mitochondria
were seen to surround the tumor cell
nucleus and empty their electron-dense material irough the nuclear membrane while malignant cells

were cured without apoptosis. Consequently "Watchful waiting" even as

active surveillance is unethical since
no attempt is made to stop CaP from
becoming more aggressive. In a case
found by screening the Gleason score
declined from 8 - 4 in six years. Multiple bone mtastases regressed in a
patient while strontium was administered, now without recurrence for
over 18 years. Hormone refractory
(HRPC) states have been evaded by,
"short time" intermittent LHRH treatment (3.6 mg Zoladex + Androcur), in
synergy with CaP powders. Further
hormone injections have been possible to be extended to intervals of 3 - 6
months. Prognostic features were defined based on uctuations in FSH,
DHEAS, PRLand PSA-levels. Additional clinical cases will be described
in detail. Conclusions: This biological non-invasive therapy is an altemative to our standard treatment modalities as it does not cause grave side-effects and deterioration of the patients'
life-quality. Activated transformed
mitochondria are involved in the healing. Improved diagnostic tests should
decrease the need for excessive
( 1 2 - 24) biopsy cores. If the Gleason
score is 7 - 10. bloody biopsies may
actually spread malignant cells into the
circulation and cause the high incidence of recurrent disease (30 - 40%)
following prostatectomy. Screening
for CaP is important since there is
now a realistic possibility to stop the
progress of CaP in the early phase of
the disease. The clinical effect of dietary supplementation, activating adrenal zona reticularis cells, can rapidly be evaluated using standard laboratory tests, and the prognostic management of prostate cancer assessed.

7th Intemational symposium on trace elements in human: new perspectives, October 13th - 15th 2009. Athens, Greece

Selenium in radiation oncology

- an update
O. Micke', L. Schomburg^,
J. Buentzel\ K. Kisters" and
R. Muecke^
'Department of Radiotherapy and
Radiation Oncology, Franziskus
Hospital, Bielefeld, ^Institute for
Experimental Endocrinology,
Charit Berlin, 'Department of
Otolaryngology, Sdharz Hospital
Nordhausen, ""Department of Internal
Medicine, St. Anna Hospital, Heme,
and ^Department of Radiotherapy,
Lippe Hospital Lemgo, Germany
(on behalf of the German Working
Group Trace Elements and
Electrolytes in Oncology (AKTE))
Tbe essential trace element Selenium, wbich is a crucial cofactor in
the most important endogenous antioxidative systems of the human body,
attracts more and more attention of
lay and expert groups. The interest of
oncologist mainly focuses in tbe
following clinical aspects: Radioprotection of normal tissues, radiosensitizing in malignant tumors, antiedematous
effect, prognostic impact of selenium
and effects in primary and secondary
cancer prevention. Selenium is a constituent of the small group of selenocysteine-containing selenoproteins
and elicits important structural and
enzymatic functions. Selenium deficiency has been linked to increased
infection risk and adverse mood
states. Se has been shown to possess
cancer-preventive and cytoprotective
activities in both animal models and
humans. It is well established that Se
has a key role in redox regulation and
antioxidant function, and hence in
membrane integrity, energy metabolism and protection against DNA
damage. Recent clinical trials showed
tbe importance of selenium for clinical
oncology. Our own clinical study involving 48 patients suggest that selenium has a positive effect on radiation-associated secondary lymphedema
in patients with limb edemas as well
as in the head and neck region, including endolaryngeal edema. Another
randomized Phase III study of our
group was performed to examine the
cytoprotective properties of selenium
in radiation oncology. The aim was to
evaluate, whether sodium selenite is
able to compensate a preexisting selenium deficiency and to prevent radiation induced diarrhea in adjuvant ra-

diotherapy for pelvic gynecologic

malignancies. By this study, for the
first time, the significant benefit of sodium selenite supplementation with
regard to selenium deficiency and radiotherapy induced diarrhea in patients with cervical and uterine cancer
has been shown in a prospective randomized trial. Disease free survival
and overall survival data implies tbat
supplementation with Se does not interfere witb the positive biological effects of radiation treatment, and might
constitute a valuable adjuvant therapy
option especially in marginally supplied individuals. More recently there
were emerging concerns coming up
from two large clinical prevention trials (NPC, SELECT) that selenium increases the possible risk of developing diabetes Type 2. Despite obvious
flaws of both studies and good
countearguments, a controversial debate remains on the possible advantage and risks of selenium in cancer
prevention. However, in the light of
the recent clinical trials, the potential
benefits of selenium supplementation
in tumor patients are undeniable even
so further research is needed.


Selenium substitution during

radiotherapy in head and neck
J. BntzeI'^O.
M.GlatzeP\U. Schfer'^
D. Riesenbeck\ K. Kisters''\
F. Bmns^\ K.G Schnekaes",
H. Dawczynski^ and R. Mcke'*'*
'Department of Otolaryngology,
Head Neck Surgery, SdharzKrankenhaus Nordhausen gGmbH,
Nordhausen, -Department of Radiotherapy, Franziskus-Hospital,
Bielefeld, -Department of Radiotherapy, Zentralklinikum Suhl
GmbH, Suhl, "^Department of Radiotherapy, Klinikum Lippe-Lemgo,
Lemgo, "^Department of Radiotherapy, Prosper-Hospital, Recklinghausen, ^Department of Internal
Medicine I, St Anna Hospital,
Heme, ^Department of Radiotherapy, Special Oncology, Medical
School Hannover, Hannover,
"AKTE, Tbe German Working
Group "Trace Elements and
Electrolytes in Oncology", and
^Pharmaceutical company biosyn,
Fellbach, Germany
Objective: The substitution of selenium effects in activation of the selenium dependent enzyme glutathione
peroxidase is important for scavenging free radicals. Until today only limited data was available about the clinical impact of selenium regarding tbe
toxicities due to free radical producing therapies, e.g. irradiation or chemotherapy. Material and methods: 39
patients (8 female, 31 male) witb advanced head/neck cancer were included in a randomized Phase II
study. The mean age was 63.52 9.31
years. Tumor localizations: oral cancer 15 patients, oropharynx 19 patients, hypopharynx 5 patients, CUP 1
patient. Group A (n ^ 22) received
500 |ig sodium selenite at the days of
radiotherapy and 300 ^g sodium selenite at holidays or weekend. Group B
(17) was irradiated without any selenium substitution. Both groups were
well balanced according age, gender,
localization and stage of the tumor.
We evaluated the RTOG grade of
radit ion-associated toxicities once
per week. Results: We observed the
following serious toxicities (Group A
versus Group B): dysphagia 22.7%
vs. 35.3%, loss of taste 22.7% vs.
47.1%, dry mouth 22.7% vs. 23.5%,
and stomatitis 36.4% vs. 23.5%. A


statistical trend {Fisher's exact-test) is

only seen in the area of loss of taste
(p = 0.172). The analysis per week
(Student t-test) had shown a significant reduction of dysphagia in the selenium group at the last week of irradiation. Conclusion: The small randomized trial has shown limited effects of selenium in the prevention of
ageusia (loss of taste) and dysphagia
due to radiotherapy hecause of head
and neck cancer.

Our results indicate that tobacco
smoking during pregnancy affected
pro-hepcidin levels in serum of mothers and their newboms. Low concentrations of some iron markers in umbilical cord blood suggest that mother's
smoking could lead to subclinical iron
deficiency in fetus.

Androgenetic alopecia and

trace element status in fertile
Tobacco smoking during pregnancy affected Pro-Hepcidin
levels in serum
M. Chelchowska', J. Ambroszkiewicz',
J. Gajewska', T. Maciejewski- and
T. Laskowska-Klita'
'Department of Newborn Screening,
and -Department of Obstetrics and
Gynecology, Institute of Mother and
Child, Warsaw, Poland
Maternal smoking is associated,
probably through a hypoxic effect on
the fetus, with stimulation of fetal
erythropoiesis and with increased fetal iron requirements. Recently as a
central mediator of iron metabolism, a
pcptidc hormone produced by the hver
known as pro-hepcidin has emerged. It
is known that anemia and hypoxia suppress hepcidin mRNA expression. The
effect of tobacco smoking on serum
concentration of pro-hepcidin and
some iron parameters in pregnant
women and their newboms were studied. Healthy matched-maternal cord
pairs (n ^ 60) were divided into nonsmoking and smoking groups according to questionnaire declaration. In
the smoking group, lower concentrations of pro-hepcidin in serum of
mothers and in cord blood were observed. It amounted respectively tto
only 85% (p < O.OI) and 70% (p <
0.0001) ofthat found in non-smoking
pairs. In both studied groups, levels of
pro-hormone in serum of women correlated positively with that in cord
Mood (r - 0.68, r = 0.54, p < 0.05).
The smoking mothers' total iron concentration was 20% lower than in
nonsmoking ones. In cord blood of
their children not only total iron but
also fenitin and transferin concentrations were decreased by 20%, 30%
and 13% respectively in comparison
to the tobacco abstinent (p < 0.05).

M.G Skahiaya and V.P. Tkachev

ANO Centre for Biotie Medicine,
Moscow, Russia
About 50% of women under 60
years old suffer from androgenetic alopecia (AA). A study of hair morphometry, clinical and laboratory parameters of blood and hair in 153 AA
women 16 - 45 years was carried out.
32 women with no alopecia manifestations were used as control. All the
women were not administrated by
trace element containing medicines
before and during the study. The
women were subjected to clinical blood
assay, hormone assay and multielement
ICP-MS hair analysis. It was found
that density of main hair (> 30 jam in
diameter) in AA women was lower
than in healthy subjects (265 3.4 vs.
320 2.5, p < 0.001). In AA group,
20 3.4% of hair in parietal zone was
fuzz-like vs. 120.5% in control. AA
women were also characterized by increased level of androstendion (24%
vs. 18% in control, p < 0.001), DHT
(47.7% vs. 25%, p < 0.001) and prolactin (3.9% vs. 0%). Multielement
hair analysis showed no significant
difference between elemental content
of parietal and occipital hair both in
AA and control groups with the exception of copper in AA women; in
this group copper level in occipital
hair was higher (16.3 0.4 \ig/g vs.
13.1 0.6 [ig/g in parietal hair, p <
0.001 ). On the other hand, within the
group of AA women, androstendion
level was found to be connected with
differences in hair elemental profile:
increased androstendion was associated with higher Cu in occipital hair,
lower Mn, Zn, Se (p < 0.05), and imbalance in Cu/Mn, Cu/Zn ratios.
Thus, the main markers for early diagnostics of androgenetic alopecia in
women are low hair density, low hair

diameter in parietal zone comparing

to occipital zone, high proportion of
fiazz-like hair in parietal zone. To reveal peculiarities of mineral exchange
in androgen-dependent and androgen-independent parts of scalp, and
for its subsequent correction, hair elemental content is advisably to be investigated simultaneously in parietal
and occipital zones of scalp.

Content of heavy metals in

thyroid glands of patients
diagnosed with nodular goitre
A. Blazewicz, G Orlicz-Szczesna,
A. Prystupa, S. Sivsammye,
A. Deol, R. Randhawa
Medical University of Lublin, Chair
of Chemistry, Department of
Analytical Chemistry, Lublin,
A nodular goitre is the most common pathology of the thyroid gland.
According to epidemiolgica! data
the annual incidence of a nodular goitre is 29 cases per 100,000 individuals. In order to study the role of a particular element in thyroid pathology
development it is necessary to undergo the comparative examinations
in healthy subjects and well-diagnosed patients. The aim of this study
was to examine the correlations between the content of selected metals
in nodular goitre (66 subjects) and
healthy human thyroid tissues (50
subjects), ion ehromatography (IC)
method was applied for the simultaneous determination of Fe' , Cu-^,
Ni^% Zn-\ Co-% C^\ Mn-- and Pb^^
in human thyroid samples, after applying microwave-assisted high pressure digestion system. Abifunctional
ion-exchange column and two complexing eluents conjointly post-column
derivatization and absorbance detection allowed quantitative determination ofthe listed metal ions separated
in isocratic conditions. The results
obtained in the present study reveal
some quantitative alterations concerning the content of certain metals
in healthy and pathological human
thyroid glands. The present work also
presents the procedure necessary to
determine the basic validation parameters of determining the transition
metal ions with the use of the IC

7th International symposium on trace elements in human: new perspectives, October 13th - 15th 2009, Athens, Greece

The significance of blood,

urine and hair multielement
analysis in estimation of
human elemental status
A.V. Skalny and M.G Skalnaya
ANO Centre for Biotic Medicine,
Moscow, Russia
The estimation of buman elemental status is a very important diagnostic problem because of differences in
reflection of macro and trace elements metabolism in different diagnostic biosubstrates (laboratory samples). The comparative study of simultaneous determination of ca. 20
macro and trace elements in scalp
hair, whole blood and urine of practically healthy persons was conducted
using ICP-OES/ICP-MS. Positive
correlations between the majority of
trace elements, Ca and Mg were
found, but negative correlations in the
case of K and Na in human hair and
wbole blood samples were found. The
strongest correlations were observed
for heavy metals. The anthropomtrie
and physiological parameters of investigated individuals correlated much
better to whole blood macro and trace
element concentrations as compared
to bair content, probably because of
strong homeostatic regulation. So, simultaneous determination of macro
and trace elements in different diagnostic biosubstrates provides more
proper information about elemental
status of humans, useful for nutritional, ecological and physiological

Hair trace element profiles in

autistic children
T.G Vizel' and A.A. Skalny^
'Research Institute of Psychiatry and
^People Friendship University of
Russia, Moscow, Russia
Examined clinically and laboratory were 38 children (30 boys and 8
girls) with autism (varying degrees of
severity) and 38 healthy children (20
boys and 18 girls) aged 2-10 years.
ICP-MS analysis of autistic children's
hair sbowed the decreased content of
Se (0.36 0,3 vs. 1.05 0.2 mcg/g in
control, p < 0.001 ), Ni (0.23 0.02 vs.
0.49 0.10 mcg/g, p < 0.001), Al
(10.820,84vs. 18.0 I.9mcg/g,p<

0.001), As(0.080.01 vs.0.470.27

mcg/g, p < 0.001 ), Cd (0.09 0.01 vs.
0.3 0.18 mcg/g, p < 0.01) and a tendency to reduce the level of Mg, Cu,
Zn demonstrated. However, children
with autism have an elevated hair
level of V (0.1 0.01 vs. 0.07 0.01
mcg/g, p < 0.05) and a tendency to an
increase in Sn (0.29 0.06 vs. 0.19
0.03 mcg/g) and Hg (0.44 0.09 vs.
0.28 0.05 mcg/g) content. When
comparing the relative performance
(frequency of deviations fi-om the
rules on the content of chemical elements in bair), found that in children
with autism, a low content of copper
(< 10 ppm) was found in half tbe cases
(44.7%), whereas in the control group
- only in 2.63%. The upper physiological hair Hg level (the hypothesis
on the etiological significance of Hg
excess in autism is actively discussed
in scientific literature) was found in
only 4% of autistic children vs. 0% in
control group. So, the obtained data
suggests, that trace element imbalances can be involved in ethiopathogenesis of autism.


mass fraction of Li (milligram per kilogram of dry bone) in rib-bone of females, males, and females and males
taken together were as follows:
0.0360 0.0024, 0.0376 0.0020,
and 0.0369 0.0015, respectively. No
clear correlation was found between
the Li content and age and gender. A
comparison of the Li mass fraction in
the rib-bone witb the Li whole blood
mass fraction for reference of men
shows that the bone accumulates at a
significantly higher (almost in 50
times) level than blood does. TTie results obtained may serve as indicative
normal values for the lithium content
in human rib-bone.

Structural speciation and

biological activity of binary
and ternary compounds of
insulin mimetic vanadium with
physiological substrates
A. Salifoglou
Department of Chemical Engineering,
Aristotle University of Thessaloniki,
Thessaloniki, Greece

The effect of age and gender

on the lithium content in rib
bone of healthy humans
S. Zaichick', V. Zaichick',
V. Karandashev', S. Ermidou-PoUet*
and S. Pollet^
'Northwestern University, Chicago,
IL, USA, ^Medical Radiological
Research Centre of Russian Academy of Medical Sciences, Obninsk,
'Institute of Microelectronics Technology and High Purity Materials,
Chemogolovka, Russia, and
''University of Athens, Athens,
Information on chemical elements
of human bones and of their body burdens may be useful for assessing nutrition and for prevention and control
of various disease states caused by
mineral or trace element imbalance.
Therefore, we studied the effect of
age and gender on the lithium content
in rib bone of healthy humans. Contents of Li in intact rib-bone of 80 relatively healtby 1 5 - 5 8 years old
women (n - 38) and men (n = 42)
were detennined by inductively coupled plasma atomic mass spectrometry. Mean values (M SEM) for the

Vanadium has been widely known

for its insulin mimetic activity. The
key to understanding tbe specific
role(s) that vanadium plays in such a
biochemical process, leading up to
the uptake and catabolism of glucose,
is the interaction chemistry emerging
between the biologically relevant oxidation states V(1V,V) and low as well
as high molecular mass molecules in
biological fluids. Prompted by that
need, the aqueous synthetic chemistry
of V(IV, V) with the low molecular
mass physiological binder a-hydroxycarboxylic (citric and malic) acids was
investigated. A pH-dependent synthetic strategy was employed on the binary V([V,V)-citric acid and the ternary V(V)-citric or malic acid-H2O2,
ultimately leading to the isolation of a
family of binary and ternary pHstructural variants for specific biologically relevant oxidation states of vanadium. The analytical, spectroscopic
and structural characterization of tbe
isolated complexes in the solid state
and in solution point out unique similarities and differences among the various anionic complexes, such as
[V^O^Li]'" (n = 2,4,6), [V'^2O2L2](n - 3,4) and [V^2O2(O2)2L2]'' (n =
2,4,6). Chemical reactivity studies



combined with in vitro biological culture work on all of these compounds

project a) to them as competent participants ofthe requisite aqueous speciation schemes, b) project the chemical
reactivity of vanadium, at different
oxidation states, toward biotargets
linking its insulin mimetic activity
with specific, albeit ill understood, biological processes of glucose metabolism, and c) contributes significantly
to the understanding of the physicochemical profiles that characterize
potentially bioavailable species of vanadium poised to promote insulin mimetic activity associated with Diabetes mellitus.
Acknowledgments: The authors
would like to acknowledge the financial support to this project by a
"FENED" grant co-financed by the
E.U.-European Social Fund (75%)
and the Greek Ministry of Development-GSRT (25%).

Blood samples were taken on Day 8

after the first inoculation. Increased
MDA and CAT, reduced SOD and
GPx and Se content, and hypovitaminoses A, C and E in the infected
chicks, compared to the healthy controls were observed. The changes in
the small intestine, lesion scores and
oocyst index, and body weight were
indicative for a severe E, acervulina
infection. Inorganic Se supplementation increased vitamin E and Se content and pGPx-activity, and improved
body weights but it did not influence
the parasitolgica! indices.

Selenoenzymes iodothyronine
A complex molecular machinery to specificaliy incorporate
selenocysteine into proteins
important for health and
L. Wurth, A. Takeuchi,
M. Rederstorff, A. Leseare,
C. Amang and A. Kro!

Antioxidant status in Eimeria

Acervulina infected chickens
after dietary selenium
N.V. Georgieva', M. Gabrashanska-,
V. Koinarski' and S. Ermidou-Pollet'

(a protein discovered in the laboratory) develop congenital muscular

dystrophies. We found that the protein resides in the endoplasmic reticulum and is highly expressed at early
stages of embryonic development. In
an attempt for a gene correction strategy, we successfully established an ex
vivo approach to restore expression
of a full-length selenoprotein N in
cultured patient-derived skin fibroblasts.

Architecture et Ractivit de !'ARN,

Universit de Strasbourg, CNRS,
!nstitut de Biologie Molculaire et
Cellulaire, Strasbourg Cedex, France

The major biolgica! form of selenium is the 21st amino acid selenocysteine. It sets apart from the other
' Faculty of Veterinary Medicine,
amino acids because of its peculiar
Trakia University, Student's Cambiosynthesis pathway and incorporapus, Stara Zagora, 'Institute of
tion into selenoproteins in response to
Experimental Pathology and
a reprogrammed UGA codon. SelenoParasitoiogy, BAS, Sofia, Bulgaria
proteins constitute a family of oxidaand 'Medical School, University of
tion-reduction enzymes involved in a
Athens, Greece
variety of well-established important
The effect of sodium selenite on
biological functions, such as reductheantioxidantstatus in broiler chick- tion of reactive oxygen species, thyens infected with Eimeria acervulina roid hormone matia^tion, sperm matwas studied. Antioxidant status was uration and muscle development. A
measured via blood plasma malonyl complex machinery, comprising at
dialdehyde (MDA), serum glutathione least eight gene products, participate
peroxidase (GPx), erythrocyte catalase in this process. The essential compo(CAT) and Superoxide dismutase nents are the SECIS element, an RNA
(SOD) activities and plasma levels of stem-loop structure in the 3' untransselenium (Se), vitamins A, C and E lated region of selenoprotein mRNAs,
with respect to parasitolgica! and and the SECIS-binding protein SBP2.
production traits. The study was per- In addition to identifying the SBP2
formed on 100, 20-day old broiler amino acids and SECIS bases imporchickens. Four groups of chickens tant for SBP2-SECIS RNA complex
were formed: the 1st - control, the formation, we discovered that the as2nd - control and received Na^SeO^ sembly of selenoprotein mRNAs with
(0.3 mg/kg diet for 10 days), the 3rd - SBP2 and other factors important for
infected with 3 >< 105 sporulated E. recoding the UGA codon requires an
acerx'uUna oocysts and the 4th - in- adaptor protein and the Hsp90
fected with E. acefvidina oocysts and chaperone complex. Patients carrying
received the same dose N
mutations in the selenoprotein N gene

S. Pavelka'^
'Institute of Biochemistry, Faculty of
Science, Masaryk University, Brno,
and -Department of Radiometry,
Institute of Physiology, Acad. Sei.
CR, Prague, Czech Republic
We describe in general properties
of selenoenzymes iodothyronine
deiodinases (IDs) of Types 1, 2 and 3
(Dl, D2 and D3, respectively) and
their fianctions in the metabolism of
iodothyronines (thyroid hormones).
In particular, we describe several applications of our newly developed radiometric enzyme assays for determination of Dl, D2 and D3 activities in
specific samples of biological material. These assays were used to follow
the alterations in IDs activities caused
by short-time incubation of cultured
astroglia! ce!ls in a chemically defined medium with different concentrations of several effectors, such as
retinoic acid, adrenergic and purincrgic agonists, substances modulating
intracellular Ca^"^ concentration, etc.
Further, it was used to determine
changes in IDs activities in various rat
tissues as a response to the effects of
some exogenous factors, such as
subchronic treatment of the animals
with ami depressant fiuoxetine. With
the aid of our new enzyme assays, we
also measured the activity of D3 in
several depots of murine white adipose tissue to fo!low development of
dietary obesity in mice fed a high-fat
diet, compared to standard !ow-fat
Acknowledgements: This work
was supported by the Academy of
Sciences ofthe CR (Research project
No. AV0Z50110509), by the Ministry
of Education of the CR (Research

7th intemationai symposium on trace elements in human: new perspectives. October 13th - 15th 2009, Athens, Greece

project No. MSM0021622413) and

by the Czech Grant Agencies GA CR
(Grant No. 304/08/0256) and GA AS
CR (Grant No. KJB401630701 ).

Protective effects of selenium,

zinc or their combination on
cadmium-induced oxidative
J. El Heni', I. Messaoudi^,
M. Banni', K. Said- and A. Kerkeni'
'Unit de Recherche, Elments
trace radicaux libres, systmes
antioxydants et pathologies
humaines et environnement. Facult
de Mdecine, Monastir, -Unit de
Recherche, Gntique, Biodiversit
et Valorisation des Bioressources,
Institut Suprieur de Biotechnologie
de Monastir, and 'Unit de Recherche: Biochimie et Toxicologie
Environnementale. ISA,
Chott-Mariem, Sousse, Tunisia
A major aim of our study is to
evaluate the potential benefit of combined treatment with zinc (Zn) and selenium (Se) in reversing cadmium
(Cd) induced histological changes
and oxidative stress in some tissues
(liver, kidney and erythrocytes). compare to Se or Zn treatment alone in rats
exposed to Cd. For this purpose, male
rats received either, tap water. Cd,
Cd+Zn, Cd+Se or Cd+Zn+Se in their
drinking water, for 35 days. The expression analysis of the two gene
classes MTl and MT2 in the liver by
means of quantitative RT-PCR in the
liver was also reported. Light microscopic examination indicated severe
histological changes in the two organs
under Cd influence. Se or Zn partially
alleviated the damage observed in the
liver. The same effect was remarked
in the kidney with Se, but no differences in the renal histological structure have been observed between the
Zn-Cd and the control groups. With
Se and Zn simultaneous treatment
during Cd exposure, the observed
morphological changes had practically disappeared from the liver, but
were only reduced in the kidney.
Marked alterations of antioxidative
system were found in Cd-treated rats.
With Se or Zn administration during
exposure to Cd, only partial corrective effects on Cd-induced oxidative
stress in the studied tissues have been

observed, while Se and Zn together

assured a more efficient protection
against the observed oxidative stress.
The molecular assay showed a decreasing trend of MT-1 relative gene
expression levels in animals supplemented with Zn, Se and their combination when compared to Cd-treated
animals. Changes of the MT-2 expression were recorded in all conditions, although fold induction levels
were less pronounced than MT-1
ones. Results demonstrated beneficial
effects of combined Se and Zn treatment in Cd-induced oxidative stress
in kidney and suggest that Se and Zn
can have a synergistic role against Cd

Action of Zn(li) and seienite on

mitochondriai function: a
comparison with Cd^^ and Ca^^
E.A. Belyaeva
Sechenov Institute of Evolutionary
Physiology and Biochemistry of
Russian Academy of Sciences,
St. Petersburg, Russia
Mitochondria are found to be target organelles for many environmental pollutants, including heavy metals; however, molecular mechanism(s) accounting for most of the biological effects of such heavy metals
as cadmium are not well understood.
Not long ago we have shown that Cd-^
induces both necrotic and apoptotic
death in different types of cells that is
accompanied by increased formation
of reactive oxygen species (ROS) at
the mitochondriai respiratory chain
complex III level and opening of mitochondriai permeability transition
pore. There is also a lot of evidence
about protective effects of zinc and
selenium, which are essential trace elements for animals and humans,
against the heavy metal-induced cytotoxicity in vitro and in vivo. Besides,
our recent data indicates that high
concentration of Zn-" or seienite are
strongly cytotoxic, producing oxidative stress and mitochondriai dysfunction via the respiratory chain disturbance, mitochondriai membrane
potential dissipation and the permeability transition induction. In tbe
present work we continued to study
molecular mechanism(s) underlying
the heavy metal-induced mitochon-


driai dysfunction and compared the

effects of Zn-* and seienite with those
of Cd- and Ca-* on isolated rat liver
mitochondria energized by different
types of respiratory substrates. Using
selective electrodes, fluorescent
probes, swelling technique and various incubation media we revealed
similarities and differences in the action
of the abovementioned metals on the
mitochondriai ftinction. Sequences of
events manifested in the mitochondriai dysfunction produced by these
metals are discussed as well. This
study was supported by the Russian
Foundation for Basic Research (grant
No. 07-04-00722 to E.A.B.)

Zinc, copper and magnesium

status in heaithy Tunisian
S. Sfar', A. Jawed^ H. Braham\
S. Amor^, F. Laporte* and A. Kerkeni'
'Unit de recherche: Elments trace,
radicaux libres, systmes
antioxydants et pathologies
humaines. Facult de Mdecine de
Monastir, Monastir,-Service de soins
de sant de base, direction rgionale
de la sant de Mahdia, Mahdia,
'Laboratoire d'analyses biologiques,
hpital universitaire "Tahar Sfar",
Mahdia, Tunisia, and ^Dpartement
de biologie intgre, centre
hospitalier universitaire de
Grenoble, Grenoble, France
Vitamins, minerais and trace elements play an important role in the
prevention of many age-associated
diseases and in maintaining normal
immune and cognitive functions. Zinc
is a structural constituent of many proteins, hormones and hormone receptors. Copper is a potent antioxidant
involved in cellular defense against
oxygen free radicals; whereas magnesium is expected to be involved in inhibition of the membrane fat including poiyunsaturated acids. For a
better understanding of the infiuencing parameters on these elements" statuses during aging and their related
metabolisms it is worthwhile to establish normal values in healthy populations. Hence, minerals and trace elements concentration can vary with
age, gender, dietary intake or even
ethnic origins. The objective of the
present study is therefore to establish


age related variations of zinc, copper

and magnesium in the healthy Tunisian population. The study included
194 healthy Tunisian subjects (99
women and 95 men) aged between 27
and 82 years old. All subjects were in
good health condition and without
chronic disease or functional impairment, according to inclusion criteria
defined for the current investigation.
Zinc, copper and magnesium concentrations were assayed in both plasma
and erythrocytes using flame atomic
absorption spectrometry method. The
obtained results showed a significant
age-related decrease in mean zinc and
magnesium total concentration (respectively for women and men, p <
0.01 : zinc (r = -0.45; -0.55), magnesium (r = -0.21; -0.31)). Furthermore, main variations are obtained in
erythrocyte levels. Mean erythrocyte
zinc level has been shown in fact to be
lower in aged subjects (over 60 years
old) (women/men: 1I.5 1.77/11.22
2.28 mg/l) in comparison to adults
(aged between 27 and 55 years old)
(women/men: 15.44 3.28/14.39
2.8 mg/1). The same variations have
been obtained in mean magnesium
erythrocyte concentration for both
women and men: 41 7.44/38.5
8.18 mg/1 and 45.93 6.11/47.21
7.08 mg/I respectively for elderly
subjects and adults. No significant
difference has been shown in copper
concentrations according to age or
gender in both plasma and erythrocytes. Sign! ficant decrease in zinc and
magnesium erythrocyte concentrations for aged subjects can be explained by age-dependence of food
absorption, some enzyme activities or
their related metabolisms. Different
aspects of trace elements metabolisms (dietary intake, blood concentration., zinc or magnesium-dependant enzyme activities...) should be
therefore investigated in the same
population and then related to health

Affects setenium indeed
concentrations of serum
autoantibodies TPOAb and
J. Kvicala', P. Hrd', V. Zamrazil',
J. Nemecek', M. Hill' and V. Jiranek'Institute of Endocrinology, and
'DataPro, Praha, Czech Republic
Many factors take part in the development of autoimmune thyroiditis
(AIT) - predominantly the genetic
predisposition, but also environmental etiologic causes. One of them
might be selenium. Selenium (Se) is
an essential component of enzymes
with an extensive regulatory and protective effect in an organism. Immunological effects of Se are documented and are distinct even above
concentrations necessary for maximal
activity of selenoenzymes. Effect of
supplementation by 100 ^g of yeastbound Se on concentrations of thyroid autoantibodies (Ab) TPOAb and
TgAb in the starting group of 349 seniors living in the Asylum Houses of
South Bohemia was monitored in the
course of three years. Se was analyzed
by Instrumental Neutron Activation
Analysis (INAA), Abs by ELISA,
both at the beginning of the trial and
after each year. Se increased from
59.9- 146.0 (Ig Se/I serum in supplemented group during the first year,
with subsequent maintaining of this
level during the next two years. Statistical evaluation of results in groups
regardless of increased Ab by ANOVA
manifested significant decrease of
TPOAb and TgAb in the course of the
first year. Decrease of TGAb continued while concentrations of TPOAb
increased during the next two years in
both supplemented and non-supplemented group. Changes were time-related. Evaluation of seniors with increased Ab revealed a similar time-dependent course but TPOAb was marginally affected by supplement. Recent results of possibility to affect serum concentration of TPOAb by Se
proved effect only for high TPOAb
concentrations. From this point of
view, it seems necessary to conduct
long-term trials with the patients with
different levels of Abs.
The work was partly supported by
grant IGA M2 CR No. NR/9160-3.

Mitochondria are criticaity

invoived in Cd (il)-induced
E.A. Belyaeva, T.V. Sokolova and
Y.A. Vlasova
Sechenov Institute of Evolutionary
Physiology and Biochemistry of
Russian Academy of Sciences,
St.-Petersburg, Russia
The present study was conducted
to elucidate the role of mitochondria
in Cd--induced neurotoxicity. We
showed that afler 3 h incubation, 500
)j.M Cd'" already produced a twofold
decrease in the viability of neuronlike PC 12 cells compared to control.
100 (JM Cd- ' became effective after 5
h incubation, whereas 50 \iM Cd2+
induced the same decrease in the cell
viability (close to 25 %) only after 24
h incubation with the cells. 10 jaM of
Cd' was not effective at all times of
incubation used. Besides, Cd'* evoked
dose- and time-dependent changes in
intracellular reactive oxygen species
(ROS) formation and respiration of
PC 12 cells. In particular, 100 and 500
(iM of Cd- produced a twofold increase in the ROS production in 30
min of incubation with the cells. After
3 h, all concentrations of the metal under test already produced an enhanced
intracellular ROS generation. However, after 24 h, 10 ^M Cd'" did not produce any significant changes in tbe
ROS formation while 50 - 500 \xM
Cd-^ sharply decreased the ROS production compared to control. In addition. Cd-', beginning from 10 ^M and
already at short incubation time (3 h),
produced a significant inhibition of
CCCP-uncoupled respiration of
PC 12 cells, i.e. the respiration with
maximal rate limited only by efficiency of the respiratory chain. An inhibition of the uncouplcr-stimulated
respiration rate is considered a marker
of the disturbance of mitochondria!
respiratory chain components. In our
hands, its complete inhibition was
reached after 3 h incubation of PC 12
cells with 500 ^iM Cd-* and after 24 h
incubation with 100 \xM Cd-\ In opposite, the restoration of the uncoupled respiration rate up to the control
level took place in the presence of 10
HM Cd-' after 24 h of incubation. At
last, we found that against Cd'-induced ROS-dependent cell injury not
only well-known antioxidants, such
as N-acetylcysteine, vitamin E, butyihydroxytoluene, and the mitochon-

7th International symposium on trace elements in human: new perspectives, October 13th - 15th 2009, Athens, Greece

drial permeability transition pore inhibitors - cyclosporine A, bongkrekic

acid and Ru-360 were effective but
also the mitochondrial respiratory
chain effectors such as CCCP and
stigmateltin (complex III inhibitor).
We concluded that reactive oxygen
species, mitochondrial respiratory
chain and mitochondrial permeability
transition pore are as the key players
in Cd (ll)-induced neurotoxicity. This
work was supported by the Russian
Foundation for Basic Research (grant
No. 07-04-00722 to E.A.B.)

Effects of cadmium and lead

ions on protein synthesis in
mouse organs
J. Sulinskiene', R. Bemotiene',
I. Sadauskiene' ^ L. Ivanoviene^
A. Liekis' and L. Ivanov' 'Institute for Biomdical Research,
and ^Department of Biochemistry,
Kaunas University of Medicine,
Kaunas, Lithuania
The present study was conducted
to investigate the effect of Cd and Pb
ions on the total protein synthesis in
mouse organs as well as on the rate
and the level of translation in cell-free
system from mouse liver. Two weeks
of mice intraperitoneal treatment with
CdClj solution resulted in the opposite effect on protein synthesis in mice
tissues - a decrease by 26% in kidney
versus 27% activation in the liver.
Meanwhile, protein synthesis in the
mouse heart was at the control level.
Mice treated with Pb(CH3COO)2 solution of the same duration had a decrease of protein synthesis in kidney
and heart by 32% and 54% respectively. Both the rate and the level of
translation were determined in cellfree systems containing S30 fraction
isolated from the liver of the following groups of experimental mice: a)
control mice; b) mice injected with
CdCl; solution for 2 weeks; c) mice
injected with Pb(CH.,C00)2 solution
for 2 weeks. These experiments
showed that Cd-^ decreased the rate of
translation by 23% and the level of
translation by 18%. Pb-' reduced
aforementioned parameters by 35%.
We also evaluated in vivo effects of
Cd and Pb ions on activity of mice liver
isoleucyl-tRNAsynthetase. Following
2 weeks of mice intraperitoneal treat-

ment with CdCb solution, the activity

of isoleucyl-tRNA synthetase was 25%
higher than in the control group. Treatment with Pb(CH3COO)2 solution activated isoleucyl-tRNA synthetase even
by 124%. This might be a part of cellular compensatory mechanisms maintaining synthesis of protein in the normal level under extreme conditions.

Oxidative activity of some iron

compounds on colon tissue
homogenates from mice after
administration of green tea,
white tea and Pelargonium
A.E. Koutelidakis', M. Serafini\
M. Komaitis' and M. Kapsokefalou'
'Department of Food Science and
Technology, Agricultural University
of Athens, Athens, Greece, and
^Antioxidant Research Laboratory,
Unit of Human Nutrition, National
Institute for Food and Nutrition
Research, Rome, Italy,
In mice, we tested the hypothesis
that iron fortifcants may exert oxidative activity on colon tissue homogenates (CTH), depending on the antioxidant capacity of infusions received with tiieir diet. CTH were obtained from mice receiving daily by
gavage 0.1 ml of infusion (8 g/100 ml
water) from green tea or white tea or
Pelargonium purpureum or catechin
(0.01 g/100 ml) or water (control) for
five days. All CTH had higher total
antioxidant capacity than the control
and lower susceptibility to oxidation
induced by the retentates of in vitro
digests of ferrous lactate, ferrous
gluconate, ferrous sulfate and
NaFeEDTA. Ferrous sulfate and
NaFeEDTA exhibited a higher oxidative effect on CTH than ferrous
gluconate or ferrous lactate. These
results are in support of our hypothesis and suggest that infusions received with diet may protect the colon from a potential oxidative effect
of non absorbed iron.


Interaction of antidepressant
drug fiuoxetine with the
metabolism of triiodothyronine
S, Pavelka'-^
'Institute of Biochemistry, Faculty of
Science, Masaryk University, Bmo,
and ^Department of Radiometry,
Institute of Physiology, Acad. Sei.
CR, Prague, Czech Republic
In the present study, we followed
the effects of subchronic administration (for 20 days) of antidepressant
drug fluoxetine, alone or in combination with 3,5,3'-triiodothyronine (T3),
on the metabolism of iodothyronines
(thyroid hormones) in the rat. In rats
treated with fiuoxetine together with
T3, we previously encountered a profound decrease in the preceding, experimentally elevated T3 plasma levels. Here, we intended to unravel this
observation. We used radiometric enzyme assays for iodothyronine sulfotransferases (ST) and UDP-giucuronyltransferase (GT), as well as our
newly developed radiometric assays
for iodothyronine deiodinases of
types 1,2 and 3 (D1, D2 and D3). The
measurements of ST and GT activities
in different fractions of peripheral tissues did not demonstrate any significant effects of the administration of
fiuoxetine, alone or together with T3,
on the induction of these enzymes.
However, treatment of rats with
fiuoxetine alone caused a moderate
increase in D2 and, in turn, a slight decrease in D3 activities in cerebellum
and some other regions of the CNS.
No significant changes in Dl activity
were detected. On the other hand, the
administration of T3 alone caused, in
accordance with our expectation, a
substantial decrease in pituitary D2
activity and a simultaneous increase
in Dl and D3 activities practically in
all tissues studied. In contrast with the
effects of T3 by itself, treatment of the
rats with the combination of fiuoxetine
plus T3 did not cause any significant
elevation of Dl and D3 activities.
Acknowledgements: This work
was supported by the Academy of
Sciences of the CR (Research project
No. AV0Z50110509), by the Ministry
of Education of the CR (Research
project No. MSM0021622413) and
by the Czech Grant Agencies GA CR
(Grant No. 304/08/0256) and GA AS
CR (Grant No. KJB401630701 ).



Zinc status (biood and hair

zinc) in pregnant Turitish
women and its reiation to
A.O. avdar'^ F. Sylemez-\
F. Aydemir% M. Baheci-,
M. Erdogan"*, I. stner* and
E. Babacan'^
'Coordinator of Trace Elements
Group (former) and Member of
Turkish Academy of Sciences
(TUBA), ^Satellite Trace Element
Center (UNESCO), 'Departments of
Obstetrics-Gynecology, 'Public
Health, and -Pediatrics of Ankara
University, Ankara, Turkey
The essential trace element Zinc
(Zn) is particularly important in periods of rapid growth including pregnancy in humans. We have been studying Zn in pregnant Turkish women for
almost three decades. In this presentation we try to summarize the results of
these studies. Serum and plasma Zn
levels measured by atomic absorption
spectrophotometry over three trimesters in 845 Turkish women (680 pregnant and 165 control ones) between
the ages of 17 to 40 years by "cross
sectional" and "longitudinal studies".
In addition, erythrocytes (RBC) and
hair Zn levels were measured in 120
of the pregnant women. They were divided into two subgroups as poorly
nourished (PN) and well nourished
(WN) ones on the basis of "nutritional
history" and "socio-economic" status
in each study. However, 290 pregnant
women in the last study were divided
into 3 nutritional groups, including
the 3rd group with "moderate nutrition". Our studies revealed lower
plasma, serum and hair Zn levels in
PN pregnant women as compared to
WN counterparts in 3 trimesters. Interestingly, plasma Zn levels were
also found to be decreased in "moderately nourished" pregnant women. In
conclusion, nutrition was found to be
the most important factor during
pregnancy in Turkey.

Iron supplementation of
fuii-term healthy breast fed
J.K. Friel, M. Moffatt and A. Miller
Departments of Pediatrics and
Human Nutritional Sciences,
University of Manitoba, Winnipeg,
Manitoba., Canada
Full-term healthy breast-fed infants have previously been shown to
benefit fiom iron supplements early
in infancy. To test this hypothesis in
another population, 82 infants were
randomized to receive 7.5 mg of supplemental iron as ferrous sulphate (1)
or placebo (P) once a day from 1 - 6
months of age. All infants were breastfed at one month of age and were encouraged to continue exclusive breastfeeding to 6 months before introducing solid foods. Blood samples were
collected by venipuncture at 4 and 12
months of age. Anthropomtrie and
demographic data was collected at 1,
4, 6 and 12 months of age when a
Bayley Developmental assessment
was conducted. Although infants
weighed more at birth (P: 3.63 0.4
vs. 1: 3.42 0.5 kg p < 0.05) this difference did not persist after one
month of age. At 4 months ferritin (1:
110 91 vs. P: 75 48) was higher
and transferrin receptors (I: 20 4.4
vs. P: 22 6) were lower in the ironsupplemented group. At 4 months
GPx was higher (I: 10.7 3 vs. P:
9.3 2) which did not persist to 12
months. Bayley MDI (P: 99 9 vs. 1:
97 10) and PDI (P: 91 14 vs. 1:
92 14) did not differ. These results
suggest that full-term healthy breast
fed infants from an advantaged population will increase iron stores when
given iron supplements with no adverse effects on growth but no advantage for developmental outcome at
one year. Supported by CIHR and

Reported intake of heme and

nonheme iron or intake of
dietary factors that can affect
iron absorption in normal,
overweight or obese children
11 - 1 2 years old in Greece
O. Malisova, G Pounis, P. Farajian,
G Risvas, A. Zmpelas and
M. Kapsokefalou
Unit of Human Nutrition, Department of Food Science and Technology, Agricultural University of
Athens, Greece
Iron deficiency is more frequent
in obese than in normal weight people. This may be related to various
physiological factors but also to dietary choices that lead to both obesity
and iron deficiency, namely foods that
are high in fat or sugars and low in
iron or other nutrients that may enhance iron absorption. There is insufficient information on the prevalence
of obesity or iron deficiency among
children in Greece or the eating habits
of children associated with the development of these problems. The objective of this study was to compare iron
intake in obese and normal weight
children. Students (n - 2553) attending the fifth or sixth grade in primary
schools in the prefectures of Attica, or
Thessaloniki, or Evia, or Viotia, or
Larissa, or Karditsa, or Rethymnon or
Chania participated to the study. Demographic and socio-economic characteristics, body mass index, daily dietary intake using a semiquantitative
food frequency questionnaire and the
trends and perception for food were
evaluated. The average daily intake of
iron was 10 10 mg iron. Children
that had high intake of iron also had
higher intake of calcium, zinc, ascorbic acid, folie acid and fiber (p <
0.001 ). Children who had low iron intake had higher BMI (p < 0.001),
higher average circumference (p ^
0.002) and higher percentage body
fat, (p ^ 0.001). The study suggests
that children that are obese may not
include in their diets foods that are
high in iron or other nutrients. Nevertheless the population tested appears
to receive adequate iron in their diets.
It may be of interest to further explore
the association between obesity, iron
intake and iron deficiency in children
in line with recent findings reported in
the literature.

7th International symposium on trace elements in human: new perspectives, October 13th - 15th 2009, Athens, Greece

The use of a modification of

the dialyzability method to
predict zinc and iron
K. Argiri, A. Kapna, E. Theofanidi,
C. Staikidou, C. Georgiou, M.
Komaitis and M. Kapsokefalou
Dept. of Food Science and
Technology, Agricultural University
of Athens, Athens., Greece
In vitro methods have been widely
used in order to predict zinc and iron
bioavailability from food and supplements. The dialyzability method, which
measures dialyzable zinc or iron released during a simulated gastrointestinal digestion as an index of their
bioavailability, has been extensively
used by many authors. Recently, we
have developed a new setup for the application of tlie diaiyzability method
that uses 6-well plates and a ring insert that holds the dialysis membrane
with the objective to increase efficiency and to allow testing small-volume samples. A series of solutions
and recipes that have been employed
in studies on zinc or iron absorption
published in the literature were digested in vitro using the new setup.
Dialyzability was measured and correlated with published values on absorption. Zinc or iron dialyzability
measured with the proposed setup
correlated well with zinc or iron absorption published in the literature.
These results suggest that the new
setup can be employed in the future
applications for the prediction of zinc
or iron bioavailability.

Influence of selenium species

and age on selenium status in
the course of selenium
J. Kvicala', V. Zamrazil',
J. Nemecek' and V. Jiranek^
'Institute of Endocrinology and
-DataPro, Praha, Czech Republic
Selenium (Se) is essential trace element with many regulatory and protective fiinctions - to mention some of
them: antioxidative protection; detoxification of carcinogens and heavy metals, control of metabolic pathways for
biosyntheses of teucotrienes, prostaglandins, prostaeyclins, thromboxanes.

immunomodulation, regulation of
thyroid hormones metabolism and effects. The aims of the study was 1 ) to
compare the effect of supplementation by various Se species (yeastbound Se and selenite) on the Se status (serum Se, urine Se, daily urine Se
excretion., urine Se/creatinine ratio)
of seniors from 3 houses for seniors in
South Bohemia and 2) to compare effect of biologically bound Se on seniors and adults. Serum Se was analyzed by neutron activation analysis
(NAA) and urine Se by fiuorimetry,
both analyses with the use of biological standards. Analyses before the intervention trials proved low Se status
and intake both in seniors and adults.
Curves obtained by statistical evaluation of analyses proved different effects of organically and inorganically
bound Se. Higher increase of serum
Se in the course of intervention by
yeast-bound Se agrees with the theory
of partial nonspecific binding of
selenomethionine (semet) to any protein and the increase of serum Se concentration. Low urine Se concentration and mainly low urinary Se excretion in the group supplemented by selenite does not fit well with the theory
of increased excretion of inorganic
forms of Se by urine. Differences in
the curves of all measured and calculated Se indices in the trial of yeastbound Se intervention of seniors and
adults are indicative of differences in
regulatory mechanisms for Se metabolism in adults and seniors.

Daily intakes of minerals and

heme iron from meat-based
dishes typical of Italian
culinary tradition
M. Lucarini, L. D'Evoli,
P. Gabrielli, A. Aguzzi and
G Lombardi-Boccia
Istituto Nazionale di Ricerca per gli
Alimenti e la Nutrizione, Roma,
The daily intake of meat in Italy is
of 130.6 g/day and bovine meat is the
most consumed meat in Italy, it (beef
plus veal) accounts for 39.4% of the
total meat and meat products consumption. Meat consumption also
greatly contributes to the daily intakes
of minerals and trace elements in the
diet; about 45% of the zinc intake.


19% of iron intake and 18% of the

Copper intake in the Italian diet is
provided by meat [1]. Furthermore,
meat represents the main source of
heme iron, the main source of highly
available iron in the diet and to get a
correct prediction of iron bioavailability from diets strictly depends on
the carefijl determination of their
heme iron content. Therefore having
data on minerals and trace elements of
cooked meat-based dishes offers reliable information on their true intake
and can represent a proper and useful
tool in getting correct calculation of
the actual nutrient intake at consumer
level and guides consumers and nutritionists in the formulation of more accurate diets. Nevertheless data on the
concentration of these nutrients in
meats-based dishes are limited. This
study was designed for providing data
on minerals and heme iron content in
meat-based dishes among the most
widespread ones in Italy. The recipes
selected were typical of culinary
preparations based on bovine meat
(^beef and veal): in pan. hamburger,
pizzaiola, cutlet, meat ball, escalope,
saltimbocca, broiled, roasted with bacon, fillet with green pepper, stew,
vitel tonne. Preparation and cooking
methods of food, especially for complex meals, lead to a modification of
Its basic composition. Minerals and
trace elements content varied depending on the ingredients utilized (ham,
parmesan, egg, discretionary salt). Ingredients induced significant increases in Ca and Na concentration
compared to raw meat. Total iron content did not show marked differences
in most of the meat based dishes compared to raw meat. By contrast losses
in heme-iron concentration were detected depending on the severity of
heating treatments. Our findings suggested that heme-iron. because of its
important health aspects, might be a
useful index of nutritional quality of
cooked meats. Finally, some micronutrients can be used as a biochemical
index predictive of the nutritional
quality of cooked foods. In this study
data on heme-iron content in meat
based dishes pointed out that hemeiron concentration varied greatly
among the recipes analyzed and that
the knowledge of the degree of hemeiron degradation in cooked meats is
determinant for accurately predicting
the level of iron availability.



Neutron activation anaiysis for

assessing trace element status
of orange juices
C. Turra', E.A. DeNadai
Femandes', M.A. Bacchi',
G.A. Sarris" and A.E. Lai Reyes^
'Nuclear Energy Center for Agriculture (CENA), University of Sao
Paulo (USP), -College of Agriculture "Luiz de Queiroz" (ESALQ),
University of Sao Paulo, and
'Informatic Center "Luiz de
Queiroz" (CIAGRl), University of
Sao Paulo, Piracicaba, SP, Brazil
Tbis work focuses on the determination of trace elements in orange
juices produced in Brazil in both conventional and organic systems. A
comprehensive sampling was performed in organic certified and conventional farms in the state of Sao
Paulo producing the sweet orange variety Valencia. Also different types
(whole, reconstituted and frozen concentrated) of commercial oranges
juices were purchased in the retail
market of Sao Paulo state. Sample
preparation consisted in freeze-dryingthejuices for the chemical characterization accomplished by instrumental neutron activation analysis
(INAA). The trace elements Br, Ca,
Co, Cs, Fe, K, La, Na, Rb and Zn
could be detennined in all samples.
The data was modeled statistically by
ANOVA and MANOVA. Br content
was significantly (p < 0.05) higher in
organic orange juices, acting as a
discriminator of production systems.
The reconstituted orange juices had
significant differences (p < 0.08) between the two production systems for
Ca, Fe and K, with higher contents in
organic orange juices. Considering
the contents of Na, the orange juices
not industrialized are healthier than
the commercial orange juices.

Conditions of the natural environment components according to ecological - geochemical monitoring and population
health data (Tomsk region,
L.P. Rikhvanov', N.V. Baranovskaya',
A.M Betyaeva', L.V. Zhomyak',
A.V. Talovskaya', O.A. Denisovaand E.G Yazikov'
'Tomsk Polytechnic University, and
^Siberian State Medical University,
Tomsk, Russia
Scientific research in the Tomsk
region has been conducted by the staff
of the Geoecology and Geochemistry
Department, Tomsk Polytechnic University. The objects of the following
research in ecological geochemical
zoning of Tomsk region are soil, solid
precipitate of snow samples (dustaerosol formations of snow surfaces),
salt formations from crockery; (scale),
as well as children hair and human
thyroid gland. In addition, there is
sufficient information of such natural
environment sources as peat, lake
bottom sediments, tree age-rings which
are referred to as sfratified formations
indicating the chemical component
intake dynamics and further retrospective evaluation of natural environment transformations in time. Tbe
scientific research shows an intensive
flux of specific radionuclides and
other elements in the natural environment of the southern Tomsk region
during the second half of the 20th century (for example, U and Pu). Many
natural and anthropogenic characteristic factors influence microelements
accumulation level in the research
material. Data anaiysis shows specific
technogenic impact in oil and gas recovery regions (Sb, Br), coal and nuclear energy enterprises (La, U, Th).
Special attention was paid to the following fact - significant U and La accumulation level in children hair and
significant uranium accumulations in
thyroid gland pathology of the population. Ranging administrative areas on a
parameter of the general complex environmental contamination, and hygienic
ranging of territory of area on degrees
of a pressure of a sanitary-and-hygienic
situation and interrelation of these parameters with the population's health is

Element composition of
human organs and tissue
N.V. Baranovskaya, L.P. Rikhvanov,
T.N. Ignatova, I.A. Matveenko, GP.
Sandimirova and N.N. Pakhomova
Tomsk Polytechnic University,
Tomsk, Russia
In the present state of biogeochemical investigations in a man, the
problem of quantitative determination of maximum possible concentration for chemical elements and their
ratios in human organs, tissues, and
organism both in normal and in pathologic state, has become utterly urgent.
Of no less importance is the revealing
of regional, national, sex-age differences in their accumulation. To
solve this problem we have attempted
to study chemical composition of a
man's organs and tissue living in
Eastern Siberia. Application of the
two high-precision methods of analysis: instrumental neuron-activation
and induction-connected plasma and
permits for determination of accumulation levels and characteristics of
distribution over the human organs
and tissue - inhabitant of Tomsk region
- 56 chemical elements. Distribution of
these elements in human organism is
extremely irregular and their variations
in concentration range up to several orders. Element accumulation in human
organs and tissues follows the general
geochemical laws of Klark and OddoGarkins for distribution of chemical
elements in the Universe. The accumulation level of elements in human
organism is determined by not only
biochemical and biophysical properties of their inctions in living tissue
but also sexual and presumably age
characteristics, pathological changes
as well as the factors of natural environment of a man. Information on accurate quantitative element composition of a man, in terms of the regional
and some other characteristics, could
contribute to more efficient correction of human health and prophylaxis
of different diseases.

7th International symposium on trace elements in human: new perspectives, October 13th - 15th 2009, Athens, Greece

On the problem of rare earth

element distribution in human
organs and tissues (by the
example of the inhabitants of
Tomsk Region)
T.N. Ignatova, N.V. Baranovskaya,
L.P. Rikhvanov and I.A. Matveenko
Tomsk Polytechnic University,
Tomsk, Russia
At present (XXI century) the problem of accurate quantitative element
structure of the human organism is
still rather topical. For this reason in
the given investigation we focus on
the processes of accumulation and
distribution of rare earth elements
(REE) in human organs and tissues.
Examination of migration, distribution and concentration of rare earth elements is impossible without having
an idea of the relation between chemical, physical-chemical properties of
these elements and biochemical
behavior of the organism. Biotic samples presenting removed pieces of human organs and tissues selected for
postmortem examination from the
men died as a result of accidents
served as a material for our study. In
general, having studied the REE distribution in organs and tissues, one
can conclude that when entering human organism, REE are involved into
a number of complex reactions. Having analyzed the results obtained, it
was revealed that accumulation of
REE spectrum occurs in the female
ovary, spleen, and adrenal gland, and
in male skin, bronchi, and lungs. It
should be noted that the maximum
concentrations of the studied elemental group are typical. First of all these
are organs of breath system that can
be explained by the rare earth element
capability to form complexes and precipitate directly in the given system,
and this system is like a sponge where
elements including rare earth elements are accumulated. It is also of
particular importance that smoking is
a factor provoking and contributing to
REE concentration in breath system.
On the whole, accumulation of rare
earth elements in human organs and
tissues follow, the general geochemical taws of Klark and Oddo-Garkins
of chemical element distribution in
the universe.

Lead in the renal calculi

GS. Pintilie (Popescu)', L. Simoiu\
I. Ionescu^ I. Gergen',
M. Harmanescu-, D. Bordean',
A. Preda^ and A. Ienciu^
'Faculty of Food Products Technology, ^Faculty of Agriculture, Banat's
University of Agricultural Science
and Veterinary Medicine Timisoara,
Timisoara, ^University of Craiova,
Faculty of Chemistry, Department of
Phisical-Chemistry, Craiova,
"National Center for NeuroMuscular Diseases "Dr. Horia
RADU", Valcele, and ^National
RandD Institute for Cryogenics and
Isotopic Technologies - ICIT
Rm.-Valcea, Valcea, Romania
Heavy metals contents in the renal
calculi might refiect a level of contamination in human environments
because it is well know that heavy
metals such as lead, cadmium, copper,
and zinc is absorbed from the intestine and excreted in the urine and even
bile through the urinary and respectively biliary tracts. The present study
is focused on the concentration of
lead in human renal calculi. In the
present study a total number of 85 urinary calculi was investigated. The
samples of renal calculi (extracted
surgically and spontaneously eliminated) were collectedfromthe adult patients of different sex and age admitted
to the Clinic of Urology Timisoara. We
established their type according to the
qualitative composition, by Fourier
transfer infrared spectroscopy. In the
next step, we investigated the distribution of lead in calculi of welldefined composition by means of
atomic absorption spectroscopy. The
environment influences the lead concentration in kidney stones. The lead
concentration is considerably increased
in the case of polluted environment.

Why are our agricultural products facing zinc deficiency?

M.j. Malakouti
Soil Science Department, Tarbiat
Modares University, Tehran, Iran
Zinc (Zn) deficiency is widespread in plants, animals, and humans
in the Middle East countries including
Iran due to the soil calcareousness.


high pH, low organic matter, coarse

texture, continuous drought, high levels bicarbonates in the irrigation water, imbalanced application of fertilizers and overuse of P-fertilizers. Zinc
deficiency is common in agricultural
products where bread and rice are the
main staples of the people. Zinc is one
of the essential elements for plants,
animals and humans, but it is deficient
(less than 1.00 mg kg ' DTPA-extractable Zn) in most calcareous soils
and consequently in plant, animal and
human diets. Zinc acts like a traffic
policeman in plants and humans.
Based on the analysis of more than
10,000 soil samples collected from
most provinces of Iran, results revealed that more than 90% of the cultivated calcareous soils are Zn deficient. Deficiency of Zn in soils on
such a large scale, and thus in plant
foods, has been suggested to be one of
the major causes of the widespread
occurrence of Zn deficiency in animals and humans. The absence of
Zn-fertilizers causes low levels of absorption of metal elements by crops. It
causes substantial yield losses in different crops and forages, and eventually disturbs animal and human
health. The importance of Zn for crop
production was recognized only during the past decade although the importance of Zn in human nutrition was
recognized much earlier. The effects
of Zn deficiency on cereal production
in Iran and Turkey have only received
attention since the early 1990s. Application of Zn-fertilizers to the calcareous soils with zinc deficiency problems has been associated with improved yield and crop quality for cereals, com, sorghum, beans, forages and
the enhancement of domestic animals
and human health.

Trace elements accumulation

in the soils in the area of Narva
power plants (Estonia)
L. Bityukova
Institute of Geology at Tallinn
University of Technology, Tallinn,
Estonia is a unique country in the
world where about 95% of electric
and thermal power is generated from
oil shale. Long-term oil shale combustion at two Narva (Baltic and Esto-



nian) power plants poses significant

negative effect on the environment.
Emission of fmest fly ash with flue
gases into the atmosphere can be a potentially significant source of atmospheric discharge of many trace elements including hazards in the soils.
Geochemical mapping of topsoils in
the densely populated Narva region
was carried out in order to evaluate
the soils contamination in the area of
Narva Power Plants. 93 topsoils collected using a regular square grid were
analyzed by the ICP-MS for 54 major
and trace elements. The spatial distribution of elements concentrations in
soils in the Narva region was estimated and the local anomalies of the
content of chemical elements in soils
on the studied area were revealed. The
statistical treatment of the data and
principal component analysis (PCA)
used for interpretation of geochemical data allow us to reveal tbe elements associations which concenfrations in the soils are generally controlled by lithology (Al, Fe, K, Ti, La,
Ni and V, P, As, Mn, Mo), and elements group (Cr and Cu) which contribution in the soils could be, at least
partially, caused by atmospheric deposition fk)m Power Plants.

Accumulation of heavy metals

in Arundo and SaUx energy
plants treated with pig slurry,
municipal sewage sludge and
inorganic fertilizers
L. Simon', B. Kovacs-, S. Bama',
C. Varga' and Z. Dinya'
'Department of Land Management
and Rural Development. Technical
and Agricultural Faculty, College of
Nyregyhza, Nyiregyhza and
^University of Debrecen, Centre of
Agricultural Sciences. Institute of
Food Safety, Quality Control and
Microbiology, Debrecen, and
^Agricultural and Molecular
Research Institute, College of
Nyiregyhza, Nyiregyhza. Hungary
A pot experiment was set up to
study the heavy metal accumulation
in giant reed (Arnndo donax L.) energy plant. Plants were grown in uncontaminated brown forest soil, in pig
slurry (excess of Cu and Zn) or municipal sewage sludge compost (excess of Cd; Cu; Pb and Zn) amended
brown forest soil for 10 weeks. Tbe

rate of Cr or Pb accumulation was low

in all plant organs. In control cultures
0.16 |ig/g Cd, in 2.5% (m/m) pig slurry
amended cultures 0.17 |4.g/g Cd, in 5%
(m/m) pig slurry amended cultures
0.16 ^g/g Cd, and in 2.5% sewage
sludge compost treated cultures 0.21
^g/g Cd was detected. In case of Cu
these values were 8.3, 15.2, 8.2, 10.4
and 10.0 |ag/g. In shoots o Arundo
39.5, 191, 44.7, 53.9 or 58.3 ^ig/g Zn
was found. Energy willow {Salix
viminalis van gigantea) was grown in
pots for 6 weeks in uncontaminated
(Cd-0.2, Cr-10.2, Cu-I2, Pb-12.5.
Zn-36.1 mg/kg) or 10% (m/m) municipal sewage sludge compost
(Cd-2.0, Cr-20.6, Cu-91, Pb-101,
Zn-1018 mg/kg) amended brown forest soil. Cd, Cr, Cu, and Pb concentrations were similar in control or sewage sludge compost treated cultures.
Zn concenfration (47.6 ng/g) was
slightly higher in treated than in control (44.9 |ig/g) plants. Open-field experiment was organized to study the
heavy metal accumulation rate of
Arundo donax and Salix viminali.s var.
gigantea energy plants. The brown
forest soil was treated with 50 t/ha or
100 t/ha municipal sewage sludge
compost in June 2008 containing <
0,5 mg/kg Cd, 14 mg/kg Cr, 140
mg/kg Cu, 19 mg/kg Pb, and 440
mg/kg Zn. There was no significant
accumulation of Cd, Cr, Cu or Pb in
Arundo or Salix leaves at the harvest
(October 2008). In the leaves of
Arundo 16.7 ng/g (control), 20.8 ^g/g
Zn (50 t/ha sewage sludge compost
treatment) or 16.9 ^lg/g Zn Zn (100
t/ha sewage sludge compost treatment) was detected at the harvest. Although the zinc content (44.4 |ag/g)
was higher in leaves of Salix treated
with 50 t/ha of municipal sewage
sludge compost than in control (21.9
^g/g), this increment proved to be statistically insignificant. Application of
ammonium nitrate fertilizer has not
affected the Cd, Cr, Cu, Pb or Zn accumulation in Arundo or Salix leaves.
Based on these results it can be supposed, that the short term hazard of
the heavy metal accumulation in
Arundo and Salix energy plants is low,
if plants are grown in soil amended
with moderately contaminated pig
slurry, municipal sewage sludge compost or inorganic fertilizer. Tbe only
exception could be zinc, which accumulation rate was higher than that of
Cd, Cr, Cu or Pb.

Heavy metals in the hostparasite system of hares in

M. Gabrashanska', V. Nanev',
M. Anisimova', V. Ermakov- and
S. Tyutikov^
'Institute of Experimental Pathology
and Parasitology, BAS, Sofia, Bulgaria, and "V I. Vemadsky Institute
of Geochemistry and Analytical
Chemistry of Russian Academy of
Sciences, Moscow, Russia
Our study was to assess tbe accumulation of heavy metals in hares
{Lepus europeaus Pallas, 1778) and in
their helminths. The effect of endohelminths (cestodes or nematodes) on
heavy metal concentrations in tissues of
hares was established under industrial
polluted field conditions as well. Content of Cd, Pb, Co, Hg, Zn and Cu was
determined in the liver, kidney and
muscle in hares infected with Trichostrongylus retortaefarmis (Nematoda)
or with Mosgovayia pectinata (Cestoda) in comparison to their helminths.
Heavy metals were determined using
atomic absorption spectrometry. The
bioaccumulation capacity of the helminths established a bioaccumulation
factor. We found out that hosts infected with cestodes had lower levels
of heavy metals Cd and Pb than hosts
infected witb nematodes. Levels of
Co, Zn and Cu were similar in the
hosts infected with cestodes or nematodes. Higher accumulation of Zn.
Hg, Pb and Cd was found in the
cestodes compared to their host and
nematodes. Our results suggest that
the model hare - M pectinata presents the promising bioindication system to evaluate environmental heavy
metals exposure in field conditions.

Effect of vanadium in ground

water from Mt. Fuji on inhabitants in vanadium rich area in
Yamanashi prefecture, Japan.
Y. Seko, M. Togawa, and
T. Hasegawa
Yamanashi Institute of Environmental Sciences, Fujiyoshida. Japan
Ground waters from Mt. Fuji have
relatively high concentration of trace
element vanadium (about 50 ^g/I) than
those in other areas (about I |ag/l). This
vanadium comes from basalt of Mt.

7th International symposium on trace elements in human: new perspectives, October 13th - 15th 2009, Athens, Greece

Fuji. The waters are widely utilized

for tap water around the mountain,
and inhabitants are ingesting the water through drinking and cooking.
Some reports suggest beneficial
health effects ofthe ground water, especially on the diabetes. If the effect is
large enough to improve the health
condition of inhabitants, some health
indicator may be better in the area
than in other areas. In order to investigate the health condition of the area,
this ecological study was carried out.
Health check data and urine samples
of subjects with informed consent
were collected at health check venues
of Fujiyoshida city, Fujikawaguchiko
town, and Hokuto city. Fujiyoshida
city and Fujikawaguchiko town are
vanadium rich areas, and Hokuto
town is not. The total number of subjects was about 800. Blood glucose, total cholesterol, high density lipoprotein
(HDL), low density lipoprotein (LDL),
percent of body fat, visceral fat level,
body mass index were not different
among the areas both in male and female. HbAlc of female was lower in
Fujiyoshida city and Fujikawaguchiko town than in Hokuto city. Urinary
salt concentration was higher in
Fujiyoshida city than in the other region both in male and female. These
regional differences will be discussed
with references to vanadium and mortality rates of these areas.

Increased rates of fatal

diseases in Kuwait as an
aftermath of the iraqi
invasion of Kuwait
L. Hayat
Kuwait University, Faculty of
Science, Biological Science
Department, Biochemistry Program,
Safat 13060
During Iraqi invasion of Kuwait
Aug/1991,730 oil wells were burned,
that discharged 7 million barrels
(MB)/ day of incompletely combusted fuel discharged to the atmosphere.
The daily emissions were CO2 ^ 1 - 2
million Tons (MT), CO = 250 T; SO2
- 65000T, NO,; 3000T and Soot:
3000T. Poly aromatic hydrocarbons
(PAH) discharged in the air was comprised of all carcinogenic PAHs (reported by the 7 international agencies
concerned with standard measure-

ments of carcinogens). Nine MB of

crude oil was dumped into the sea.
The concentration of vanadium and
Nickel in Ku crude oil is 35 and 9
mg/kg oil successively. The poly-aromatic hydrocarhons discharged contained large quantities phenanthrene,
that can be completely ingested and
transformed in the blood, The heat of
the combustion and sea water used to
extinguish the fires oxidized the metals into their soluble salts. The soot
particle carried both the PAHs and the
minerals salts. The dominant direction ofthe wind in Kuwait N and NW
contaminated the whole environment
in Kuwait. Kuwait Institute for Scientific Research (KISR) studied % of
pollution relative to the folds of contamination in local plants and animals. The results showed that the soot
fallouts increased the levels of [V]
(mg/kg) rang in Sweet Basils and
Garden Rockets: 3 and 9 fold. The
twelve PAHs studied all increased
drastically in ehicken eggs, sheep
milk and cow milk. The fallouts on
the sea caused 10 and 5 fold elevation
in the [V] and [Ni] ofthe surface sea
soil. The increased concentrations of
minerals reflected negatively on the
marine creatures. It is reported by L.
Hayat (2009) that V and Ni deposited
in the gills; liver and flesh of benthic
organisms and fish. A study by KiSR
(1999) reported that an adult in Kuwait consumes 19.6 g/day offish and
shrimps, therefore according to L.
Hayat calculation, an adult will ingest
16.3 ng Ni/day and 24.6 |ig V/day,
plus further ingested V and Ni from
local vegetables and fruits. Inhalation
of polluted air, synergistically increased the levels of V and Ni in the
blood of patients with brain tumors.
In a span of 5 years (1990 - 1995),
brain tumors increased from 50 to 365
cases/million of population. Cancers
in general increased in Kuwait after
oil combustion and sea spillage from
0.05% to 0.1 % cases in Kuwait population. Immediately after the invasion, early maturity between girls
aged 7 - 9 years, sudden death ofthe
fetus in pregnant women, kidney failure, multiple sclerosis, diabetes; high
cholesterol levels and strokes low Vitamin D and osteoporosis increased
drastically in Kuwait population.
During the Gulf war in 1991 more
than 300 tones of D U was used by aircrafts and tanks in Kuwait and Southem Iraq over an area about 20.000


km^. Large fraction of DU bullets

(-90%) fired from aircrafts missed
their intended targets. They compromise about 250 tons of DU, buried at
various levels in the ground. With
time, chemical weathering oxidized
metallic DU into soluble chemical
forms that contaminated the soil and
ground water. It eventually incoq^orated into the human food chain. The
Gulf population was exposed to the
depleted uranium in a number of
ways, 1) exposure to the radiation,
emitted from uranium isotopes, 2) the
inhalation of depleted uranium particles, and 3) the ingestion of contaminated water or food. Systematic study
of Kuwait soil started in 1993. Samples were collected from the whole
territory (83 sites) and sediments (43
sites) from Arabian Gulf ground. The
radioactivity measurements showed
that the ratio of isotopic rate of
235(j/u23 is 20 times greater than the
nonnal range: Normal range U-'^/U^^**
- 0.03%/99.97% - 0.0003%. Du was
detected in the hair and nails of 385
Kuwaiti Citizen. The percentage and
degree ofthe contamination range was:
10% high. 60% medium and 30% low
(S. Al Hashim 2006). People exposed
to radiation of DU suffered from joint
pain, kidney failure and cancers, respiratory diseases and lung cancer, skin
diseases and genetic deformatities, and
liver cancers. Al! the above mentioned
diseases drastically increased in Kuwait
and Iraq - especially the southern part
(AL hasra).

Biomonitoring of trace elements

in the Czech population
A. Krskov, M. Cem,
V. Spevckov, J. Smid,
M. Cejchanov and K. Wranov
National Institute of Public Health,
Prague, Czech Republic
In the Czech Republic, selected
metals in human body fluids are monitored within (1) the "Czech Human
Biomonitoring Project" (CZ-HBM)
and (2) the "Public health impact of
long-term, low-level mixed element
exposure in susceptible population
strata" project (PHIME) within the
6th Framework Programme of EU. In
the CZ-HBM, toxic (lead, cadmium,
mercury) and essential (selenium,
copper, zinc) trace elements are moni-


tored in blood, urine and hair of adult

and child population. These activities
routinely run since 1994 and are divided into two periods (first period:
1996 - 2003; second period; since
2005). Selected important outputs
are: (1) higher blood lead levels in
men and boys vs. women and girls,
(2) declining trend of blood lead levels in time, (3) significant difference
in blood mercury levels between fish
eaters and fish non-eaters, (4) blood
selenium levels have slightly upward
trend but they did not reach optimal
values yet, (5) assessment and revision of reference values for different
metals and population groups [1, 2].
The PHIME project will run over five
years (2006 - 2011 ). PHIME contains
37 different work packages or subprojects. 28 of these are organized
into four different Pillars. Czech Republic is involved in Pillar III - Human Biomonitoring. Selected metals
are examined in the blood of the children, occupational ly exposed workers and postmenopausa! women. Preliminary results are: (1) slightly
higher blood lead levels in children
from the rural area (29.7 ^ig/l) in comparison with city and local-industrial
locality and the capital city (24, 25
and 25 ^g/l. respectively), (2) no child
has blood lead level higher than 100
)ig/l [3], (3) significantly higher blood
lead levels in exposed workers than
those of controls or the Czech general
population. The correlation of lead in
whole blood and plasma is still under
This work was supported by the
EU through its Sixth Framework
Programme for RTD (contract no
FOOD-CT-2006-016253). Reflects
only the author's views. The Community is not liable for any use that may
be made of the information contained


Cenia M. Spevckov V. Bettes B.

C'ejchani>v M. SmidJ. Reference values
for lead and cadmiiiin in biood of Czech
populalion, Inl J Occup Med Environ
Health, 2001;/4,- 189-192.
Bariov A. Spevckov V. Benes B.
Cejchanov M. Smid.l, Cerna M. Blood
and urine levels of Pb, Cd and Hg in the
general population of the Czech Republic and proposed reference vaiues, Int J
Hyg Environ Health. 2006; 209:


Ewers U. Kraus C. Schulz Ch. Wilhelm

M. Reference values and human biological monitoring values for environmental
toxins, Int Arch Occup Envirun Health,
1999; 72, 255-260.

Element content in children

hair samples (Tomsk Region,
N.V. Baranovskaya, D.V. Shvetsova
and L.P Rikhvanov
Tomsk Polytechnic University,
Tomsk, Russia
It is a well- known fact that human
hair is the most preferable medium for
different experimental researches, as
its composition reveals the level and
changes of those numerous miero-element traces in a human organism
over a long period of time. Hair chemical content reflects the general human organism state and, in its turn, is
widely studied to give an ecological
description of the population and as a
diagnosis for the population health
level. Human hair element content
changes in accordance to race health
level (physiological state), diets, hair
color, medicine application, cosmetics, geochemical conditions of habitat
and work-place. Human hair is considered to be an excellent indicator of the
environment influence on man i.e.
heavy metal emission (fimes), industry and heat-power plant discharge, incineration emission. Thus, the analysis of tbe human hair is an essential
moment in monitoring different environmental conditions. The research
subject was hair of children living in
Tomsk Region (N = 568). Hair samples of children from 3 - 1 5 (boys and
girls) from Tomsk oblast were investigated. For comparative analysis hair
samples from two Russian regions
and other territories- Irkutsk oblast
(36 samples), Cheljabinsk oblast (44
samples), Stolin, Belorussia (3 samples), and Pavlodar, Kazakhstan ( 120
samples) were also investigated.
Above-mentioned comparative selection is based on the following fact that
they are all considered to be environment-trouble territories (due to existing
nuclear-iel cycle plants, petrochemical, chemical, engineering plants, as
well as, three heat-power plants). The
geochemical element spectrum in the
hair of children from Tomsk oblast is

rather diverse. This indicates that the

hair element content in the population
of Tomsk region is complex and predominantly includes specific technogenic features. The specific feature of
the chemical content in children's hair
(Tomsk oblast) is the increased concentration of rare earth elements La,
Ce, Sm and radioactive elements Th,
U. There is a rather high concentration of Na, Ca, Sc, Cr, Fe, Co and also
some rare earth elements La, Ce, Hf in
hair samples of children living in
Tomsk oblast. A significant high content level of La, Lu, Sm and Th was
found in the hair samples of children
living, not only in Tomsk oblast, but
also in Cheljabinsk oblast. Such a
high concentration in hair is due to the
impact of nuclear-fuel cycle enterprise (Seversk, Ozersk). There is a
rather high U concentration in children's hair samples from Irkutsk
oblast and Pavlodar, Kazakhstan. Another important factor is the significantly low index ratio of Th to U in
these regions (Th/U = 0.06), while in
otber regions it includes 0.4 - 0.5. It
should also be noted that a high Fe
content in hair is specific for Tomsk
and Irkutsk oblasts, while selenium
content only for Irkutsk oblast and
Kazakhstan. In conclusion, Tomsk
oblast was conventionally divided
into five zones in accordance to the
technogenic environment transformations: (1) area with high technogenic environment transformations;
(2) regions with highly- developed
petroleum production; (3) Tomsk region; (4) Seversk, where the Siberian
Chemical Plant (SCP) is located; (5)
populated areas located in the northeastern direction from the Siberian
Chemical Plant. The element content
in children's hair in above-mentioned
zones showed that the concentration
of most elements increases from Zone
1 to Zone 5. However, such a tendency was characteristic only for the
following elements- Co, La, U, and,
in some cases - Cr, Sc. There is a high
Au content level in the hair of those
children living in oil regions; high Na,
Sb, Ag, Yb content in hair for Tomsk
obiast in general, while a concentration of radioactive elements (U, Th) is
found in the hair of those children living in the populated areas located in
the north-eastern zone of the Siberian
Chemical Plant.

7th International symposium on trace elements in human; new perspectives, October 13th - 15th 2009, Athens, Greece

Concentrations of metals in
welder's blood
1. Martinsone
Riga Stradins University, Riga,
In Latvia, approximately 2.4% of
the workforce is involved in metal
processing industry. According to
laboratory measurements of metal
processing industry workplace air by
Riga Stradin's university Institute of
Occupational Safety and Environmental Health Laboratory of Hygiene
and Occupational Diseases, working
environment was estimated to be of
poor quality. The aim of this study
was to investigate metals concentrations in welder's blood, because it is a
widely used biomarker in different
countries. Overall 104 welders were
sampled for metals (manganese, chromium, nickel, and zinc, copper) concentrations in blood. The concentrations of metals in blood are very varied;
welders had higher metal levels of manganese and chromium in blood than the
recommended reference interval.

Present methods for

biogeochemical indication of
microelementhoses of animals
S. Tyutikov', V. Ermakov',
M. Gabrashanska^ and
M. Anisimova'V.I Vemadsky Institute of Geochemistry and Analytical Chemistry
of Russian Academy of Sei.
Moscow, Russia, and 'Institute of
Experimental Pathology and Parasitology of Bulgarian Academy of
Sei., Sofia, Bulgaria
At the present stage of development of biological indication methods,
they are subdivided both from the
type of parameter used, and considering the organization levels observed
(e.g., molecular, cellular, organismic
or systemic). The purposes of bioindication also differ as well as its
methods. Besides its abovementioned
application in exploration geology,
nowadays bioindication approach is
widely used in ecological monitoring
of regions. There the original method
of biological indication of chronic
microelementhoses in animals through
chemical elemental composition of

hair covering was developed. The

methods that we have developed
when free-ranging wide animals or
livestock are used, are deprived the
drawbacks of the above-stated estimation methods. On the contrary,
they provide determination of environmental pollutions by toxic elements and pesticides, while enabling
differentiation of the pollutions for
their intensity and duration. A way of
integral evaluation of microelemental
ecological status for the environment
and agrocenoses with an opportunity
of subsequent correction of the microelementoses is offered.
Supported by grant of RFFR
09-05-0008 la.

About strontium and selenium

in soil of East Siberia in connection with manifestation of
Urov disease
V.V. Ermakov
V.l. Vemadsky Institute of Geochemistry and Analytical Chemistry
of the Russian Academy of Sciences, Moscow, Russian Federation
In the period of 1998 to 2007
some comparative biogeochemical
researches have been conducted in
East Zabaikalia (namely, the basins of
the Shilka, the Unda, the Gazimur, the
Urjumkhan and the Urov rivers), including both endemic and non-endemic territories. Se was determined
in soils by means of spectrofluorimetric method with 2,3-dianiinonaphtaline and Sr was detected by
AAS. High contents of Sr in soils of
Urov disease-affected endemic areas
have been observed in Proterozoic
granitoid out zone more frequently.
Now for carbonate soils of this area
Se mobility rates are sufficiently
higher, and such areas are free from
any endemic disease. Nevertheless,
parameters of Se mobility in the soils
of the Urov disease-affected area of
the East Zabaikalia are comparable to
those for some other Se-deficient areas of the Chita Region and other areas of Russia. Taking into account existence the Urov disease-affected area
with the normal status of selenium
also, the etioiogical role of the last,
apparently, is not main.
Supported by grant of RFFR
09-05-0008 la.


The possibility of reducing the

environmental risk with water
treatment sludge In heavy
metal contaminated soil
S. Bama', L. Simon' and A. Anton^
'Department of Land Management
and Rural Development, College of
Nyiregyhza, Nyiregyhza and
Research Institute for Soil Science
and Agricultural Chemistry of the
Hungarian Academy of Sciences,
Budapest, Hungary
Chemical stabilization is an effective in situ soil remediation technology for soils contaminated with toxic
metals. The water treatment sludge as a potential chemical stabilizing additive - was tested in soil incubation
models. Soil samples were collected
from the neighborhood of an abandoned lead-zinc sulfide ore mining
area (Gyngysoroszi). It was contaminated with mine spoil, which contained various heavy metals (e.g. cadmium, copper, lead, zinc and nickel).
Stabilizing substrate was given in 2 or
5% (m/m) to the contaminated soil
and the heavy metal fractions dissolved by nitrohydrochlorie acid,
Lakanen-Ervi buffer, acetate buffer
and distilled water were measured.
With the aqua regia extraction we
have determined the "pseudototal"
metal content in soil. The extraction
with Lakanen-Ervi buffer characterized the metal fraction, which is lose
in medium period, and is available for
plants. The acetate extract is available
for plants directly, while the distilled
water extract characterized the most
mobile fraction of metals in soil. Risk
assessment was made with RISC 4.0
software which is a software package
for performing human health risk assessments for contaminated sites. In
aqua regia extract there was no considerable reduction or change in risk,
since metals in water treatment sludge
are also extracted. Treatments have
not changed the heavy concentration
in "plant available" fraction, but the
cumulative environmental risks was
slightly reduced by 1.90 - 4.35%. The
stabilizing ability of the substrate was
confirmed - the water treatment
sludge decreased mobile fraction of
toxic metals in the soil, thus the cumulative environmental risk was also reduced in soil, and in groundwater,
which is potentially endangered. The
environmental risk was reduced in
case of soil by 16.94 - 40.38% for



adults and 15.86 - 40.92% for children, while in case of groundwater

this reduction was 15.15 - 41.21%
and 16.17 - 41.25%. In distilled water
the water treatment sludge application
reduced the heavy concentrations by
13.29-41.2%, while the reduction of
environmental risk in contaminated
soil was 6.14 - 11.31 %, in groundwater 5.68-11.49%.

ples. Instrumental developments of

accelerators (with a higher stability in
energy) permit us to obtain nanobeams in the order of 100 - 200 nm for
PiXE-RBS and 50 nm for STIM.

Commercial hair analysis:

science or quackery?
S. Zaichick' and V. Zaichick^

Micro and Nano-ion beam

analysis of trace elements at
biomaterials/living tissues
E. Jallot
Clermont Universit, Universit
Blaise Pascal, Laboratoire de Physique Corpusculaire de
Clermont-Ferrand CNRS/IN2P3
UMR 6533, Aubire, France
To better understand the specific
role of relevant biological trace elements, it is important to study at the
micrometer and nanometer scales
bioceramics, cells and interfaces that
bioceramics develop in vitro and in
vivo. Ion beams methods of analysis
permit us to obtain simultaneously
multi-elemental maps and concenfration measurements of major, minor
and trace elements with a spatial resolution from micrometer to nanometer.
These techniques are very useful to
determine local concentration gradients at biomaterials/cells interface.
Ion beams methods of analysis like
PIXE (Particle Induced X-ray Emission) or RBS (Rutherford Backscattering Spectroscopy) are based on interactions between charged particles
and atoms present in the samples.
PIXE method generally uses proton
beams with energies between 1.5 and
3 MeV to ionize atoms and to generate
characteristic X-rays. The limit of detection of PIXE is several ppm. A further very important advantage is that
PIXE can be used as a micro-beam
technique, which allows elemental
mapping with a good spatial resolution (1 ^m) and to quantify local elemental concentrations. Backscattered
particles spectroscopy (RBS) permits
us to obtain stoichiometric information for light elements and local
charge measurements to quantify
PIXE data. Scanning Transmission
Ion Microscopy (STIM) allow us to
obtain images of density in thin sam-

'Northwestern University, Chicago,

IL, USA, and -Medical Radiological
Research Centre of Russian Academy of Medical Sciences, Obninsk,
Disregarding art paper advertisements of the laboratories, which offer
hair "mineral" analysis, there is a
question of reliability of the "hair
mineral test" and the scientific validity of the "body mineral status" estimation using the "hair mineral test"
results. Therefore we performed the
following study with two objectives.
The first objective was to evaluate
inter- and intra-laboratory agreement
concerning hair chemical element
analysis. The second objective was to
check correlations between chemical
element contents in hair and bone tissue. Comparison to bone was made
because many chemical elements
have a high affinity to bone tissue,
thus a skeleton is a target organ for
them. Contents of chemical elements
in intact scalp hair and rib-bone of 80
relatively heaithy 1 5 - 5 8 years old
women (n ^ 38) and men (n ^ 42)
were determined by ICP-AES. Identical collections of hair samples were
analyzed in two commercial Moscow
laboratories, which offer "hair mineral" analysis today. Rib bone samples were analyzed in the Analytical
Center of the Institute of Microelectronics, Technology and High Purity
Materials. It was shown that "hair
mineral" analysis from contemporary
laboratories is unreliable. Moreover,
"hair mineral" analysis does not reflect
content of bone-seeking elements. It
means that "hair mineral" analysis
does not reflect content of boneseeking elements in the human body,
because the skeleton is the biggest
pool of such elements. We must recommend refraining from using "hair
mineral" analysis to assess a "body
mineral status", individual nutritional
status, or suspected individual environmental exposure.

Preconcentration and determination of Uranium in fertiiizers

1. Varga', N.L. Misra- and . Szles^
'Institute of Chemistry L. Etvs
University, Budapest, Hungary,
Fuel Chemistry Division, Bhabha
Atomic Research Centre, Mumbai,
India, and 'Institute of Isotopes
Hungarian Academy of Sciences,
Budapest, Hungary
Phosphate rocks, which contain
uranium 5 - 5 0 times higher concentration than uranium crustal abundance, are the raw materials of fertilizers. Uranium and its compounds
have been classified as toxic materials: So, to avoid bioaccumulation during agricultural use of phosphate containing fertilizers, efforts have been
made to increase uranium elimination
efficiency from fertilizers, for decades [1]. A remarkable increase of
uranium concentration was detected
in the plough layer of arable soils due
to long-term application of superphosphate [2]. Total reflection X-ray
fluorescence spectrometry (TXRF)
was applied for uranium determination in seawater [3] and human urine
[4]. In the present work we demonstrate that uranium content of N-P-K
fertilizers can be successfully determined by total reflection X-ray fluorescence spectrometry (TXRF). Fertilizers, manufactured in Hungary,
Germany, Austria and Denmark were
analysed applying preconcentration
methods of selective extraction and
co-precipitation. TXRF results were
compared to the data of analysis by inductively coupled plasma sector field
mass spectrometry (ICP-SFMS).





LongRS. Process for ihe recovery ofuranium from phosphatic ore, US Patent
2.882,123, 1959.
Rothbaum HP. McGaveston DA. Wall T.
Jahnston AE. Maltiiigly CEG Uranium
accutnulation in soils from long-continued applications of superphosphate. Eur
J Soil Sei, 1979; .fl,- 147-153,
Misra NL. Dhara S. Sing Mudher KD.
Uranium determination in seawater by
total reflection X-ray fluorescence spectrometry. Spectrochim Acta PartB. 2006;
61: 1166-1169.
Zarkadas Ch, Karydas AQ ParadelUs T.
Determination of uranium in human
urine by total refleclion X-ray fluorescence, Spectrochim, Acta PartB. 2001;
Jo. 2505-2511.

7th International symposium on trace elements in human: new perspectives, October 13th - 15th 2009, Athens, Greece

Assessment of 59 trace
element contents in rib bone
of healthy humans by
inductively coupled plasma
mass spectrometry

Selenium quantification and

speciation in human spermatozoa: A description of present
methods with a look at the
future development

S. Zaichick', V. Zaichick^
V. Karandashev^ and I. Moskvina^

H. Bertelsmann, D. Behne,
M. Hammadeh and
A. Kyriakopoulos

'Northwestem University, Chicago,

IL, USA, -Medical Radiological Research Centre of Russian Academy
of Medical Sciences, Obninsk, and
-'Institute of Microelectronics Technology and High Purity Materials,
Chemogolovka, Russia
The effect of age and gender on 59
trace element contents in the intact rib
bone of 80 relatively healthy 15 - 55
year old women and men was investigated by inductively coupled plasma
mass spectrometry (ICP-MS). Mean
values (M SEM) for the mass fraction of Ba, Bi. Cd, Ce, Cu, Gd, La, Li,
Mo, Nd, Pb, Pr, Rb, Sm, Sr, TI, U, and
Zn (milligram per kilogram of dry
bone) were: 2.54 0.16, 0.0153
0.0017, 0.0522 0.0071, 0.0286
0.0024, 1.35 0.22, 0.00153
0.00012, 0.0197 0.0019, 0.049
0.011, 0.11 0.03, 0.011 0.001, 2.7
0.3, 0.0032 0.0004, 1.75 0.24,
0.00016,0.0013 0.0001, and 92.8
1.5, respectively. The upper limit of
mean contents of Ag, Al, B, Be, Br,
Cr, Cs, Dy, Er, Hg, Ho, Lu, Mn, Ni,
Sb, Tb, Te, Th, Ti, Tm, Y, and Yb
were: < 0.015, < 7.2, < 0.65, < 0.0032,
< 3.9, < 0.33, < 0.008, < 0.0025,
< 0.0010, < 0.018. < 0.00053,
< 0.00024, < 0.36, < 1.05, < 0.0096,
< 0.0004, < 0.005, < 0.003, < 2.8,
< 0.00019, < 0.0047, and < 0.0008, respectively. In all rib bone samples the
contents of As, Au, Co, Eu, Ga, Hf, Ir,
Nb, Pd, Pt, Re, Rh, Sc. Se, Sn, Ta, V,
W, and Zr were under detection limits
(DL): 0.01, 0.001, 0.3, 0.0007, 0.2,
0.002, 0.0001, 0.01, 0.01, 0.002,
0.0005, 0.02, 0.1, 0.2, 0.2, 0.001,
0.03, 0.1, and 0.03, respectively. Statistically significant (p < 0.001, t-test)
tendency for the Ce, Gd, La, Nd, Pr,
and Sm content to increase with age
was found in the human rib bone, regardless of gender. It was shown that
higher Sr mass fractions were typical
of female ribs as compared to those in
male ribs.

Helmholtz-Zentrum Berlin fr
Materialien und Energie, Berlin,
Selenium in spermatozoa is linked
to male fertility. Two forms of glutathione peroxidase 4 are present in
mammalian sperm, the sperm nuclei
glutathione peroxidase (snGPx) and
the phospholipid hydroperoxide glutathione peroxidase (PHGPx). Different analytical methods including instrumental neutron activation analysis (INAA) and atomic absorption
spectrometry (AAS) were used to determine selenium in sperm. The investigation on the chemical form of
selenium in sperm is usually carried
out by immune detection on blot
membranes with an antibody suitable
for the simultaneous detection of
snGPx and PHGPx. However, this
method is in the best case a semiquantitative approach as these selenoproteins may show a different immune reactivity. For the quantification of all selenoproteins in sperm in
one experiment laser ablation coupled
to mass spectrometry or long-term irradiation with INAA, methods which
can be applied directly on gels with
separated selenoproteins, can be
useftil in future studies.

Characterization of human
selenoproteins (Sep 15 and
other) by HPLC-ICP/MS and
Specific ELISA
J. Bartel, T. Bartz, A. Sote, J. Bartel
and A. Kyriakopoulos
Helmholtz Zentrum Berlin fur
Materialien und Energie, Dept.
"Molecular Trace Element Research
in the Life Science", Berlin,
In previous decades a number of
studies established selenium as an element of fundamental importance to
human health. The trace element is a
component in selenium binding proteins and present in at least 25 selenocysteine containing proteins, summa-


rized as selenoproteins. Inadequate

supplementation has been related to
severe diseases such as cancer, rheumatoid arthritis and Alzheimer's disease while deficiency was linked to a
decrease of immune and thyroid ftinction, a loss of male fertility and a
higher susceptibility to develop viral
diseases. Selenium's occurrence within selenoproteins seems to be important for these physiological roles,
however the detailed mechanisms remain unclear. Artificial selenium deficiency by feeding a depleted diet
can help to unveil the machinery responsible for the elements essentiality
by resulting in a diminished expression of selenoproteins. Here we introduce a method suitable for the quantitative determination of selenoproteins
in human cells and cell organelles
organdes based on the separation of
tryptic peptides by HPLC and on-line
coupling to ICP/MS for the detection
of selenium. Additionally we established an enzyme-linked immunosorbent assay (ELISA) for the quantification of the 15kD-selenoprotein
and compared the results

Selenoproteins in the nuclear

A. Graebert, D. Schmidt and
A. Kyriakopoulos
Helmholtz Center Berlin for
Materials and Energy, Department
"Molecular Trace Element Research
in den Life Sciences", Berlin,
Selenium, an essential trace element in mammals, is incorporated as
selenocysteine in selenoproteins. Nutritional deficiency decreases the biosynthesis of selenoproteins comprehensive and leads to pathologic conditions. But genetic modifications can
also result in the lack of particular
selenoproteins. Alzheimer s disease,
colorectal cancer and muscular dystrophy, e.g., can be associated with genetic defects ofa selenoprotein gene.
To date about twenty-ve mammalian
selenium-containing proteins are
known, Many of the characterized
selenoproteins act in the regulation of
cellular redox processes. But the biological function of some of these proteins, especially with predicted membrane association, remains unknown.
Membranes are one of the least studied cell structures. Because they are


hydrophobic lipid bilayers and integral membrane proteins often have

pi's in the alkaline region, this inherent problem is adverse in using traditionally 2-DE technique. In this work
we separated nuclear membrane proteins from hepatoma cells by means of
an alternative electrophoretic technique for separating membrane proteins: The two dimensional 16.BAC /
SDS-PAGE. By metabolic labeling
with 75 Se and electrophoresis, the
selenoproteome ofthe nuclear membrane was analyzed. The nuclear envelope is an important cell compartment in the action of signaling and
traffic control between cytoplasm and
nuclear interior. It is known that mutations in the nuclear envelope proteins
emerin and lamin A cause a number of
human diseases including premature
aging syndromes, muscular dystrophy, and cardiomyopathy. But it is
still remained unanswered up to now
whether selenoproteins play a role in
these pathways.

Is scalp hair a valid indicator

for the assessment of lithium
content in the human body?
S. Zaichick', V. Zaichick-,
V. Karandashev', S. Ermidou-Pollef
and S. Pollet'
'Northwestern University, Chicago,
IL, USA, ^Medical Radiological
Research Centre of Russian
Academy of Medical Sciences,
Obninsk, institute of Microelectronics Technology and High
Purity Materials, Chernogolovka,
Russia, and "University of Athens,
Athens, Greece
Lithium is widely used in medicine as an antidepressive drug. Therefore we performed the following
study with two objectives. The first
objective was to determine the normal
levels and age-related changes of Li
in the scalp hair of healthy women
and men - residents of an uncontaminated area. The second objective was
to check correlations between Li contents in hair and rib-bone tissue. Comparison to bone was made because Li
has a high affinity to bone tissue, thus
a skeleton is a target organ for them.
Contents of Li in intact scalp hair and
rib-bone of 80 relatively healthy
15-58 years old women (n ^ 38) and
men (n = 42) were determined by inductively coupled plasma atomic

mass spectrometry. Mean values (M
SEM) for the mass fraction of Li (milligram per kilogram) in scalp hair of
females, males, and females and
males taken together were as follows:
0.0221 0.0035, 0.0399 0.0056.
and 0.0317 0.0036, respectively. A
tendency for an increased Li mass
fraction with age was observed in the
scalp hair of females only. It was
shown that lower Li content was typical of female scalp hair as compared
to those in male hair. No clear correlation was found between the Li content
in scalp hair and rib-bone. The results
obtained may serve as indicative
normal values for the lithium content
in human scalp hair.

Hair as an indicator of elements'

trace bioaccumulation in
human body: "optimization of
a washing procedure ofthe
hair by an endemic clay
(Rhassoul) of Morocco"
S. El Fadeli', A. Pineau- and
A. Sedki'
'Laboratoire d'Hydrobiologie,
Ecotoxicologie et Assainissement,
Marrakech, Maroc, and ^Laboratoire
de Toxicologie et d'hygine
industrielle, Nantes Cedex, France
For many years, hair analysis has
played an important role in the monitoring of toxic risks. Concentrations
of trace elements in hair reflect their
mean level in the human body. Measuring the concentrations of harmful
trace elements (Pb, Cd and At) in hair
can be used to monitor a population's
exposure to pollution or to estimate
intake from contaminated foodstuffs.
A further application is the biomonitoring of personnel occupationally
exposed to heavy metals. To ensure
the significance and reliability of results, the quality ofthe analytical procedure is paramount. Indeed, when
measuring trace element levels in hair,
only the internal (endogenous) fraction should be taken into account,
hence, the importance ofthe washing
procedure used prior to analysis. In
this context, our work explores new
ways of valorization of Moroccan clay,
locally called Rhassoul, employed traditionally for centuries for fine cosmetics. It is characterized by its absorbing
and cleaning properties as it was used
for a long time like a natural shampoo
or product of washing. The objective of

this study was to test the effectiveness

and aggressiveness of two washing
procedures to compare removal of endogenous elements and extemal contaminants. The procedure finally adopted was the one that most effectively
removed all extemal contaminants
without being too aggressive without
removing the elements incorporated
into the hair matrix during its growth,
as this fraction alone reflects the accumulation of trace elements in the organism. This study showed the effectiveness of Rhassoul in the retention
ofthe metal cations (Cd, Ph, et Al.) in
the hair.

Sensitive assay of sulfite in

environmental samples by
N-(9-acridinyl)maleimide (NAM)
M. Yamaguchi', T Kamata^ and
H. Meguro'
'Department of Health Sciences,
Tohoku Fukushi University, Sendai,
and -Jumonji University, Sugasawa,
Niiza, Japan
A highly sensitive and specific determination method for sulfite (SOr )
was developed using HPLC with
NAM fluorometry. The sulfite has
various influences on environment.
Sulfite is one ofthe common oxidized
forms of sulfur. It dissolves to a rain
drop, readily oxidized into sulfuric
acid to form acid rain. Sulfites directly cause serious damage to plants.
The final stable forms cause serious
ecological problems by increasing
acidity of the lake and pond waters.
Under anaerohic conditions, sulfite is
fonned in organic rich sediments by
biochemical reduction of sulfate by
sulfate reducing bacteria. Sulfite as a
reducing agent decreases the oxygen
content in coastal areas and natural
waters, which has a harmful effect on
aquatic life and cultivation ofthe environment. However, there is no report
conceming determination for sulfite of
the sediment origin. It should be noted
that it is difficult to determine sulfite
under dissolved oxygen. On the other
hand, sulfite and its salts are extensively utilized in various processed
foods as antioxidants and bleach.
Therefore there is a strong demand for
a simple and rapid method to determine sulfite, specifically from environmental samples. Recently, we have
reported a highly sensitive assay of

7th International symposium on trace elements in human: new perspectives, October 13th - 15th 2009, Athens, Greece

sulfite by NAM fluorometry using

HPLC. The reaction of sulfite with
NAM was successfully applied to determine sulfite in wine, serum, acid
rain and acid snow at pico mol level
by Meguro H., Akasaka K., Yamaguchi M. ( 1983 - 2006). !n this study,
we would like to report an application
of the method to determine sulfite in
marine sediment using HPLC-NAM
fluorometry. Marine sediment was
collected in Matsushima Bay. Miyagi
Prefecture Japan, in the extraction
buffer containing EDTA, which effectively prevented the loss of sulfites
by oxidation. NAM reacted with the
sulfite in marine sediment quantitatively and gave stable and strong fluorescent derivatives, The concentration range of the determination was
about 1.0 - 1000 pmol on ODS column in HPLC. The sulfite concentration of 120 - 770 ppm was detected in
marine sediment. The determination
required less than 100 mg of marine
sediment. The recovery of the added
sulfite was almost quantitative.

Extremely sensitive determination of iodothyronine

deiodinase enzyme activities
S. Pavelka'^
'Department of Radiometry, Institute
of Physiology, Academy of Sciences
of the Czech Rep., Prague, and
^Institute of Biochemistry, Faculty of
Science, Masaryk University, Bmo,
Czech Republic
Tbe aim of the present study was
to find out the proper assay conditions
and to work out reliable methods for
extremely sensitive radiometric determination of the enzyme activities
of iodothyronine deiodinases (IDs) of
types 1, 2 and 3. These enzymes catalyze selective (outer ring) 5 '- and (inner
ring) 5-monodeiodinations of iodothyronines and play, therefore, crucial
roles in the metabolism of thyroid
hormones. Our newly developed radiometric assays for IDs were based
on the use of highly specific radioactivity '"''I-labeled iodothyronines as
substrates; TLC separation under optimum conditions of radioactive products from the unconsumed substrates;
film-less autoradiography of radiochromatograms using storage phosphor screens; and quantification of
the separated compounds with a

BAS-5000 (Fujifilm Life Science

Co.) laser scanner. This methodology
enabled us to determine IDs enzyme
activities as low as 10"* katals. We
have used these enzyme assays for the
quantification of IDs activities induced in cultured astroglial cells by
several purinergic and adrenergic
agonists and agents controlling the
intracellular Ca-^ concentration, as
well as for the determination of IDs
present in microsomal fractions of
various rat tissues. The assays proved
to be very sensitive and rapid and, at
the same time, reliable and robust.


cant effect on the iodine determination result (a ^ 0.1). A good correspondence (for 10 additional determinations) between the certified (3.38
0.02 ppm with variation coefficient
(v.c.) of 0.59 % for standard reference
material (srm) and the measured iodine concentrations (3.52 0.29 ppm;
v.c. = 10 %) was achieved. The applied conditions of digestion, reinforced by the action of microwaves,
brought about a decidedly shorter
(less than 20 minutes) sample preparation time.

This work was supported by the
Academy of Sciences of the CR (Research project No. AV0Z50110509),
by the Ministry of Education of the
MSM0021622413) and by the Czech
Grant Agencies GA CR (Grant No.
304/08/0256) and GA AS CR (Grant
No. KJB401630701).

Determination of iodine ions in

human thyroid glands using
High Performance Ion Chromatography method
A. Blazewicz, G Orlicz-Szczesna,
A. Prystupa, W. Dolliver, N. Parikh
and H. Kaur
Medical University of Lublin, Chair
of Chemistry, Department of Analytical Chemistry, Lublin, Poland
High Performance Ion Chromatography (HPIC) with a pulsed amperometric detection (IC-PAD) proceeded by microwave mineralization
was applied for determination of iodides in human thyroid samples. The
aim of the study was to examine correlations between the content of iodides in nodular goiters and healthy
human thyroid tissues obtained at autopsy. A fast, accurate and precise
ic-pad method followed by alkaline
digestion in a closed system and with
the assistance of microwaves was developed and used for the comparative
analysis of the two types of human
thyroid samples. Statistical analysis
revealed over 8-foId reduction of iodine concentration in the pathological
tissues in comparison with the control
group. It was pointed out that the way
of tissue preservation (either in formalin or by freezing) had no signifi-

Effect of iron supplementation

an the iron status and the
pregnancy outcome in
pregnant women in Greece - a
prospective study
V. Papatzelou', A. Antsaklis^,
A. Protopappas', K. Sotirakoglou^
and M. Kapsokefalou'
'Department of Food Science and
Technology, Agricultural University
of Athens, ^Department of Obstetrics and Gynecology, University of
Athens, "Alexandra" Hospital and
'Department of Science, Laboratory
of Mathematics and Statistics, Agricultural University of Athens,
Athens, Greece
Iron deficiency in pregnancy is associated with consequences for mother
and baby; therefore, along with dietary
advice, iron supplementation is commonly prescribed. Its effectiveness was
investigated in 263 women. Hematocrit
(Hct) and hemoglobin (Hb) were measured bimonthly. On the 12th-15th
week of gestation, 55 women were
iron deficient (ID) (Hct < 32%) and
205 were iron adequate (IA) (Hct
> 32%) and were advised to receive
daily 200 or 100 mg iron. Women
from the ID group remained iron deficient until the end of gestation, while
women from the IA group preserved
adequate levels of iron. Area of residence or employment did not affect
the results. The iron status of the
mother did not affect the gestation period or the birth weight of the newbom. These results suggest that, in
this group, high iron supplementation
was not able to correct iron deficiency
during pregnancy and highlight the
importance of iron adequacy and
proper nutrition before entering pregnancy.

Copyright of Trace Elements & Electrolytes is the property of Dustri-Verlag Dr. Karl Feistle GmbH & Co., KG
and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright
holder's express written permission. However, users may print, download, or email articles for individual use.