10th International Magnesium Symposium Schedule of Events

INTERNATIONAL SOCIETY FOR THE DEVELOPMENT OF
MAGNESIUM RESEARCH
The 10th International Magnesium Symposium is one of a series of International
Magnesium Symposia organised every three years by the International Society for
Development of Magnesium Research (SDRM).
HONORARY PRESIDENT
Jean Durlach (France)
HONORARY SCIENTIFIC COMMITTEE
K.W. Beyenbach (U.S.A.)
H.G. Classen (Germany)
R.J. Elin (U.S.A.)
Y. Itokawa (Japan)
M.S. Seelig (U.S.A.)
Y. Rayssiguier (France)
HONORARY SCIENTIFIC ADVISORY COMMITTEE
L. Alouane (Tunisia) F.J. Alvarez-Leefmans (Mexico)
P. Bac (France) A. Cittadini (Italy)
F. Corica (Italy) I.H. De Leeuw (Belgium)
S. Djurhuus (Denmark) J.F. Escanero (Spain)
P.W. Flatman (U.K) A. Flynn (Ireland)
M.J. Halpern (Portugal) S.A. Kiss (Hungary)
A. Kobayashi (Japan) T. Kozielec (Poland)
B. Lasserre (Switzerland) A. Mazur (France)
R. Mettey (France) R. Mittendorf (U.S.A.)
H. Mori (Japan) P.J Porr (Romania)
G.A. Quamme (Canada) A.J. Reyes (Uruguay)
P.M. Rob (Germany) R.K. Rude (U.S.A.)
R. Rylander (Sweden) N.E. Saris (Finland)
K. Shivakumar (India) R.H. Smetana (Austria)
G. Stendig-Lindberg (Israel) T. Theophanides (Greece)
R. Vink (Australia) W.B. Weglicke (U.S.A.)
C.Y. Yang (Taiwan)

The 10 th International Magnesium Symposium is sponsored by:
Young Investigator Awards:
Blaine Pharmaceuticals:
The Magnesium Resource since 1955
http://www.blainepharma.com
Mineral Water:
Magnesia Mineral Water:
The power of natural magnesium
http://www.mattoni.com
Financial Management:
The University of Adelaide:
Australia's third oldest university
and known internationally for excellence
in research and teaching
http://www.adelaide.edu.au

Schedule of Events

10 th International Magnesium Symposium
Schedule of Events
SUNDAY, SEPTEMBER 7 TH
14:00 – 18:00 Registration Pre-function Area
18:30 – 20:30 OPENING CEREMONY AND WELCOMING
RECEPTION
MONDAY, SEPTEMBER 8 TH
Session 1 PLENARY LECTURES: The clinical relevance of
magnesium from gestation to old age
Chair: Dr Robert Vink (Australia)
8:30 New data on the importance of gestational Mg deficiency
Dr Jean Durlach (France)
9:00 Mental benefits of estrogen replacement therapy (ERT) in
postmenopausal women; risks of cognitive loss and
dementia intensified by ignoring importance of
magnesium
Dr Mildred S. Seelig (USA), Burton M. Altura, Bella T. Altura
9:50-10:20 COFFEE BREAK
Poster set-up (anytime after 9:00)
Poolside Terrace
Plaza Ballroom A
1
2
Pre-function
Session 2 NEUROSCIENCE
Chair: Dr Ibolja Cernak (USA)
Plaza Ballroom A
10:20 The influence of some antipsychotic drugs on erythrocyte
and plasmatic magnesium level and of other bivalent
cations
Dr Mihair Nechifor (Romania), C.Vaideanu, I.Palamaru,
C.Borza, I.Mindreci
3
10:40 Effects of magnesium on blood-brain barrier breakdown
and brain edema In septic rats
Dr Tulin Erdem (Turkey), Damla Aktan, Ayse Erturer,
Mukadder Orhan, Nahit Cakar, Figen Esen
4
11:00 Magnesium-Vit B6 intake reduces central nervous
hyperexcitability in children
Dr Marianne Mousain-Bosc (France), Roche M., Rapin J.R.,
Bali J.P.
5
11:20 Effect of magnesium on neural activities in rat cortex and
hippocampus in vitro
Dr Keiichi Torimitsu (Japan), Nahoko Kasai, Yasuhiko Jimbo,
Yuriko Furukawa
6
11:40 Magnesium gluconate offers no more protection than
magnesium sulphate following diffuse traumatic brain
injury in rats
Dr Robert Vink (Australia), Renee J. Turner, Katherine W.
DaSilva, Christine O’Connor, Corinna van den Heuvel
7
12:00-13:15 BUFFET LUNCH Poolside

Session 3 GENETICS
Chair: Dr Richard Gardner (New Zealand)
Plaza Ballroom A
13:15 Magnesium transport genes in yeast and plants
Dr Richard Gardner (New Zealand), Jong-min Lee, Keith
Richards, Salam Salih, Paul Donaldson, Van Kelly, Revel
Drummond, Ana Tutone
8
13:45 Molecular biology of magnesium transport proteins
Dr Matin Piskacek (Austria), Jochen Stadler, Martin Kolisek,
Gabor Zsurka, Julian Weghuber, Monika Schweigel, Rudolf J.
Schweyen
9
14:15 Mg2+ transport proteins of the yeast plasma membrane
and cytoplasmic homeostasis factors
Jochen Stadler, Anton Graschopf, Stefan Köstler, Dr Rudolf J,
Schweyen (Austria)
10
14:35 The pathogenesis of Machado-Josephs Disease; a high
manganese / low magnesium induced CAG expansion
mutation in susceptible genotypes?
Dr Mark Purdey (UK)
11
15:00-15:20 COFFEE BREAK Pre-function
Session 4 TRANSPORT
Chairs: Dr Masato Konishi (Japan); Dr Charles Coudray
(France)
15:20 Interest of stable isotopes use in the evaluation of Mg
metabolism
Dr Charles Coudray (France), Christine Feillet-Coudray,
André Mazur, Yves Rayssiguier
15:50 Effects of intracellular concentrations of Na+ and Mg2+ on
Mg2+ transport in rat ventricular myocytes
Dr Masato Konishi (Japan), Michiko Tashiro and Pulat
Tursun
16:10 Depletion of intracellular Mg2+ via Na+-independent
passive Mg2+ pathways
Dr Shinsuke Nakayama (Japan), Hideki Nomura, Lorraine M.
Smith, Joseph F. Clark, Tadayuki Uetani, Tatsuaki Matsubara
16:30 Set point shift of the intracellular Mg2+ concentration with
amiloride and KB-R7943 in vascular smooth muscle
Mr Tadayuki Uetani (Japan), Hamaguchi Yukihisa,
Tatematsu Yasushi, Nakayama Shinsuke
17:00-18:00 Poster Session A
17:30-19:30
Cocktails
Plaza Ballroom A
12
13
14
15
Pre -function

TUESDAY, SEPTEMBER 9 TH
Session 5 PLENARY LECTURES
Chair: Dr Robert Vink (Australia)
Plaza Ballroom A
8:30 Magnesium and the inflammatory response: potential
physiological and clinical implications
Dr Yves Rayssiguier (France), Andrzej Mazur
16
9:15 Irukandji Syndrome – another clinical role for magnesium
sulfate?
Dr Michael Corkeron (Australia)
17
9:45-10:05 COFFEE BREAK Pre-function
Session 6 YOUNG INVESTIGATORS AWARD SYMPOSIUM
(Sponsored by Blaine Pharmaceuticals)
Chair: Dr Robert Vink (Australia)
10:05 The effect of magnesium deficiency on primary tumour
growth and the effectiveness of antitumour treatment in
transplantable murine cancers
Ms Anna Nasulewicz (Poland), Joanna Wietrzyk, Yves
Rayssiguier, Andrzej Mazur, Adam Opolski
10:30 Investigation of intracellular magnesium mobilization
pathways in PC12 cells by simultaneous Mg-Ca
fluorescent imaging
Mr Takeshi Kubota (Japan), Hirokazu Komatsu, Yutaka
Shindo, Kentaro Tokuno, Yoshiichiro Kitamura, Hiroto Ogawa,
Koji Suzuki, Kotaro Oka
10:55 Magnesium attenuates post-traumatic depression/anxiety
following diffuse traumatic brain injury in rats
Ms Lisa Fromm (Australia), Deanne L. Heath, Robert Vink,
Alan J. Nimmo
11:20 The CorA-Mrs2 superfamily of Mg2+ transport proteins in
bacteria and mitochondria
Ms Elisabeth Froschauer (Austria), Martin Kolisek, Martin
Piskacek, Gabor Zsurka, Julian Weghuber, Monika Schweigel,
Rudolf J. Schweyen
11:55-12:30 STANDING SANDWICH LUNCH
12:30
AFTERNOON EXCURSIONS
Plaza Ballroom A
18
19
20
21
Poolside
Hotel Entrance

WEDNESDAY, SEPTEMBER 10 TH
Session 7 CARDIOVASCULAR I
Chair: Dr Mildred Seelig (USA)
Plaza Ballroom A
8:30 Effects of oral magnesium therapy on exercise tolerance,
exercise-induced chest pain, and quality of life in patients
with coronary artery disease Dr Michael Shechter (Israel),
C. Noel Bairey Merz, Hermann-Georg Stuehlinger, Joerg
Slany, Otmar Pachinger, Burton Silver and Babeth
Rabinowitz
22
9:00 A prospective non-randomized, open-labeled pilot study
investigating the use of magnesium in patients
undergoing non-acute percutaneous coronary intervention
with stent implantation Vladimir Rukshin, Raul Santos, Mitch
Gheorghiu, Prediman K. Shah, Saibal Kar, Sriram
Padmanabhan, Babak Azarbal, Vivian T. Tsang, Raj Makkar,
Bruce Samuels, Norman Lepor, Ivor Geft, Steve Tabak,
Mehran Khorsandhi, Neil Buchbinder, Neil Eigler, Bojan
Cercek, Keta Hodgson, Dr Sanjay Kaul (USA)
23
9:20 Optimal administration dosage of magnesium sulfate for
torsades de pointes in children with long QT syndrome
Dr Kenji Hoshino (Japan), Kiyoshi Ogawa, Takashi Hishitani,
Yoshikatsu Etoh
24
9:40 Effects of Mg2+ on cardiac excitation-contraction coupling
Dr Anouchka Mihaylova (USA), Belik M.E., McCulloch A.D.
25
10:00-10:15 COFFEE BREAK Pre-function
Session 8 CARDIOVASCULAR II
Chair: Dr Larry Resnick (USA)
Plaza Ballroom A
10:15 Intracellular magnesium assay correlations with: oral &
transdermal absorption, endothelial function, exercise
tolerance, and chronic disease depletion
Dr Burton B. Silver (USA), C. Norman Shealy, M Shechter,
Mark C.P Haigney, Allison J. Stewart, Kim Brower
26
10:45 Intracellular magnesium in furosemide treated patients
with congestive heart failure suffering from diabetes
mellitus and/or renal dysfunction Dr Irena Alon (Israel), D.
Almoznino-Sarafian, S. Berman, O. Gorelik, M. Shteinshnaider,
D.Modai, N.Cohen
27
11:05 Long-term outcome of intravenous magnesium therapy in
thrombolysis-ineligible acute mayocardial infarct patients
Dr Michael Shechter (Israel), Hanoch Hod, Babeth
Rabinowitz, Valentina Boyko, Pierre Chouraqui
28
11:25 In vitro application of endotoxin to thoracic aortas from
magnesium-deficient rats enhances vascular
hyporeactivity to phenylephrine
Dr Atsushi Miyamoto (Japan), K. Moriki., S. Ishiguro, A.
Nishio
29
11:45 Case for subcutaneous magnesium product for space
missions
Dr William J. Rowe (USA)
30
12:05-13:10 BUFFET LUNCH Poolside

Session 9 CARDIOVASCULAR III
Chair: Dr Irena Alon (Israel)
Plaza Ballroom A
13:10 Relation of hypertension and atherosclerosis: 1H- and
31P-NMR spectroscopy of free Mg, plasma membrane and
circulating lipids in hypertension
Dr Lawrence M. Resnick (USA), Raj K. Gupta
31
13:45 Functional effects of magnesium and statin
pharmaceuticals compared
Dr Andrea Rosanoff (USA), Mildred S. Seelig
32
14:05 Role of magnesium in pathogenesis of essential
hypertension Dr Genel Sur (Romania), Oana Maftei
33
14:25 Cardiovascular and skeletal benefits of estrogen
replacement therapy in healthy postmenopausal women;
risks intensified by ignoring importance of magnesium
Dr Mildred S. Seelig (USA), Burton M. Altura, Bella T. Altura
34
15:00-15:20 COFFEE BREAK Pre-function
Session 10 SKELETAL
Chair: Dr Juergen Vormann (Germany)
15:20 Osteoblastic cell growth as a function of Ca 2+ /Mg 2+ ratio
O. Theodorakopoulou, D. Deligianni, Dr Jane
Anastassopoulou (Greece)
15:50 Bone mineral density, serum albumin and serum
magnesium
Dr Noboru Saito (Japan), Naoto Tabata, Toshiaki Setoguchi,
Saburou Saito, Harumi Sayama, Toshiko Kannagi
16:20 Magnesium, magnesium deficiency and interaction with
aminoglycoside and quinolone antibiotics
Dr. Juergen Vormann (Germany)
16:50-18:00 Poster Session B
(All posters to be removed at 18:00)
Plaza Ballroom A
35
36
37
Pre-function
19:30- GALA DINNER Plaza Ballroom

THURSDAY, SEPTEMBER 11 TH
Session 11 SPORT
Chairs: Dr Stefano Iotti (Italy); Dr Frank Mooren (Germany)
Plaza Ballroom A
9:00 Magnesium in sports
Dr Frank C. Mooren (Germany)
38
9:30 Cytosolic free [Mg2+] in the human calf muscle in
different metabolic conditions
Dr Stefano Iotti (Italy)
39
10:00-10:20 COFFEE BREAK Pre-function
Session 12 MEASUREMENT IN RELATION TO PUBLIC HEALTH
Chair: Dr Kay Franz (USA)
Plaza Ballroom A
10:20 A functional biological marker is needed for diagnosing
magnesium deficiency
Dr Kay B. Franz (USA)
40
10:50 The relation of birth weight to intracellular magnesium of
cord-blood platelets
Dr Junji Takaya (Japan), Fumiko Kotera and Yohnosuke
Kobayashi
41
11:15 Balance of Mg positively correlates with that of Ca
Dr Mamoru Nishimuta (Japan), Naoko Kodama, Eiko
Morikunii, Yayoi H. Yoshioka, Hidemaro Takeyama, Hideaki
Yamada, Hideaki Kitajima and Kazumasa Suzuki
42
11:40 Developed determination method of ultra-trace elements
and ultra-trace element levels in plasma of rat fed low
magnesium diet
Dr Mikeo Kimura (Japan), Kazuto Honda, Atsuko Takeda,
Masayo Imanishi, Takahisa Takeda
43
12:00-13:00 BUFFET LUNCH Poolside
Session 13 OTHER APPLICATIONS TO CLINICAL MEDICINE
Chair: Dr Federica Wolf (Italy)
13:00 Magnesium and cancer in clinical practice (update).
Mr Amid Reba (France), François Goldwasser
13:25 Intracellular magnesium is independent from extracellular
availability during proliferation
S. Fasanella, A. Torsello, B. Tedesco, A. Cittadini, Dr
Federica Wolf (Italy)
13:45 Magnesium, insulin resistance and body composition in
healthly posmenopausal women
Maria Jose Laires (Portugal), Moreira H, Monteiro CP,
Sardinha L, Veiga L, Gonçalves A, Bicho M
14:05 Clinical efficacy of magnesium supplementation in
patients with type 2 diabetes Dr K.Yokota (Japan), M.Kato,
H. Ii, M. Shiraishi, T. Hayakawa, T. Kikuta, F. Lister, H.
Akiyama, S.Kageyama and N.Tajima
14:25 Post-cholecystectomy syndrome and magnesium
deficiency
Dr Paul J. Porr (Romania), J. Szántay, M. Rusu
14:45 CLOSING CEREMONY
YOUNG INVESTIGATOR AWARDS
FAREWELL DRINKS
Plaza Ballroom A
44
45
46
47
48
Plaza Ballroom A

POSTERS Plaza Ballroom
Pre-function area
Effect of magnesium diets in ischemic stroke rats
49
Céline Demougeot, Sylvie Bobillier Chaumont, Claude Mossiat, Christine
Marie and Alain Berthelot.
A substance P antagonist increases brain intracellular free
50
magnesium concentration after diffuse traumatic brain injury in rats
Robert Vink, Maria I. Cruz, James J. Donkin, Alan J. Nimmo and Ibolja
Cernak
Amiloride increases neuronal damage after traumatic brain injury in
51
rats
Renee J. Turner, Corinna van den Heuvel and Robert Vink
Propofol attenuates the neuroprotective effects of magnesium in
52
experimental traumatic brain injury
Tulin Erdem, Damla Aktan, Mehmet Kaya, S.Murat Imer, Nahit Cakar, Lutfi
Telci, Figen Esen
Simultaneous monitoring of extracellular magnesium and zinc in
53
gerbil cortex during focal cerebral ischemia by microdialysis-graphite
furnace atomic absorption spectroscopy
FU-Chou Cheng
Determination of extracellular magnesium in brains of gerbils
54
subjected to cerebral ischemia by an on-line microdialysis and
graphite furnace atomic absorption spectrometry
Jen-Bin Lee
Microdialysis coupled with graphite furnace atomic absorption
55
spectrometry in the determination of blood magnesium levels in
gerbils subjected to cerebral ischemia/reperfusion
Ming-Cheng Lin
Contribution of the Na+-Mg2+ exchanger on insulin-induced
56
modulation of the intracellular Mg2+ concentration in rat hearts
Yasushi Tatematsu, Shinsuke Nakayama, Tetsuya Amano, Kenji Imai,
Manabu Kokubo, Tadayuki Uetani, Takaaki Yamada, Yukihisa Hamaguchi,
Toyoaki Murohara, Tatsuaki Matsubara
A study on spontaneously obese rats (Minko rat) with abnormal lipid
57
metabolism -strength and mineral concentrations in bone
Ryuji Takeda, Takashi Nakamura, Masayo Imanishi, Madoka Ishida,
Fumiko Yano, Takahisa Takeda, Mieko Kimura
Modulation of tyrosine kinases and phosphatases by Mg2+ ions in
58
human red blood cells
Alexander Barbul, Yehudit Zipser, Nechama S. Kosower, Rafi Korenstein
Is determination of Mg pool size useful to assess Mg status
59
C. Feillet-Coudray, C. Coudray, E. Gueux, A. Mazur and Y. Rayssjguier
Effects of reduced magnesium availability and mild oxidative stress
60
on aging of WI-38 human diploid fibroblasts
B.Tedesco, S. Fasanella, A.Torsello, A.Cittadini and F.I. Wolf.
Food intake and magnesium intake affect true absorption and
61
endogenous fecal excretion of magnesium in rats
Hideyuki Ohmori, Tohru Matsui and Hideo Yano
Serum magnesium levels and dependency/disability in hospitalised
62
elderly patients
M Barbagallo, LJ Dominguez, A Ferlisi, A Galioto, A Pineo, C Aglialoro
Absorption and effect of the magnesium content of a mineral water in 63
the human body
Sándor A. Kiss, Tamás Forster, Ágnes Dongó,
About the misdiagnostics of magnesium deficiency
64
Dierck-H.Liebscher, Dierck-E.Liebscher.

Magnesium in asthma attack
G. Sur, N. Miu, Lucia Burac, Oana Maftei.
Experimentally induced prolonged magnesium deficiency causes
osteoporosis in the rat
Stendig-Lindberg G, Koeller W, Bauer A, Rob PM
Modifications of the plasmatic and salivary magnesium
concentrations and of other bivalent cations in patients with
suppurations of the oro-maxilar area
Nechifor Mihai, Gradinaru Irina, Mîndreci Ioan, Tatarciuc Monica,
Gogalniceanu Dan
Lyme disease and magnesium deficiency
V. Cristea, M. Crian and V Crian
Effect of magnesium on essential oil formation of genetically
transformed and non-transformed chamomile cultures
Szokee, É., Máday, E., Kiss, S.A. And Lemberkovics, É.
Magnesium in animal nutrition
Katalin Kovacsne Gaal, Orsolya Safar, Laszlo Gulyas, Petronella Stadler
Mg-content of Rhizobium nodules in different plants and the
importance of Mg in N2-fixation of nodules
Kiss S.A., Stefanovits-Bányai É., Takács-Hájos M.
65
66
67
68
69
70
71

1
New data on the importance of gestational Mg deficiency
Jean Durlach. Society for the Development of Magnesium Research, Neuilly-sur-
Seine, France. Jean.Durlach@wanadoo.fr
Chronic primary Mg deficiency is frequent. Around 20% of the population consumes
less than two-thirds of the RDA for Mg, in both genders and in women in particular.
For example, in France, this applies to 23% of women and 18% of men. Primary Mg
deficiency may intervene in fertile women. Gestational Mg deficiency may induce
maternal, fetal and pediatric consequences that may last throughout life.
Experimental studies of gestational Mg deficiency have shown that Mg deficiency
during pregnancy may have marked effects on the processes of parturition and
postuterine involution. It may intervene on fetal growth and development from
teratogenic effects to morbidity, i.e. hematological effects and disturbances in
temperature regulation. Clinical studies on the consequences of maternal primary
Mg deficiency in women have been insufficiently investigated. To check the validity
of the role of this frequent gestational Mg deficiency, the protocol of a long-term,
multi-center, placebo controlled, prospective study on the effects of maternal
nutritional Mg supplementation on lethality and morbidity in the fetus, neonates,
infants, children and adults should be carried out not only during pregnancy and the
first year of life, but throughout life.
Two forms of chronic gestational Mg deficiency have been stressed: (i) premature
labor when chronic maternal Mg deficiency is involved in uterine hyperexcitability,
and (ii) Sudden Infant Death Syndrome (SIDS) when it is caused by either simple Mg
deficiency or various forms of Mg depletion.
(i) Nutrional Mg treatment of premature labor. If gestational Mg deficiency is the
only cause for uterine overactivity, nutritional Mg supplementation constitutes the
etiopathogenic atoxic tocolytic treatment. But though it is an adjuvant factor in
premature labor, it is only a useful accessory treatment, devoid of toxicity but which
increases effectiveness and safety of the associated tocolytic drugs such as beta-2
mimetics.
(ii) SIDS due to gestational Mg deficit: Mg deficiency or various forms of Mg
depletion? SIDS may be caused by the fetal consequences of the maternal Mg
deficiency through an impaired control of brown adipose tissue (BAT)
thermoregulation, mechanisms leading to a modified temperature set point. SIDS
may result from dysthermias: hypo- or hyperthermic forms. A possible prevention
could rely on simple nutritional Mg supplementation. Various stresses in pregnant
women or in the infant may transform a simple Mg deficiency into Mg depletion:
stress in baby care such as bedding in the prone position and environmental factors
such as parental smoking. But the role of chronopathological stress in particular
appears to be too often neglected as it constitutes a clinical form of primary
hypofunction of the biological clock (with its anatomical and clinical stigma such as
reduced production of melatonin (MT) and of its urinary metabolite, 6 sulfatoxylmelatonin
(6-MT). SIDS might be linked to an impaired maturation of both the
photoendocrine system and BAT. The preventative treatment of this form of SIDS
should associate atoxic nutritional Mg therapy for pregnant women with total light
deprivation at night for the infant. The place of Mg therapy for the infant and of MT,
L-tryptophan and taurine is uncertain yet.

2
Mental benefits of estrogen replacement therapy (ERT) in postmenopausal
women; risks of cognitive loss and dementia intensified by ignoring
importance of magnesium
Mildred S. Seelig, Burton M. Altura, and Bella T. Altura. Department of Physiology
and Pharmacology, State University of New York, Downstate Medical Center,
Brooklyn 11203, USA. mgseelig@comcast.net
Better cognition and lower incidence of Alzheimer’s dementia among
postmenopausal women receiving ERT than in those not so treated, is explicable by
ERT’s normalization of brain activation patterns in some women performing verbal
and figural memory tasks. Memory, learning, encoding, retrieval, reasoning, and
other higher cognitive functions were superior among many women receiving
estrogen. Cessation of estrogen secretion is associated with losses in those
capacities. Estrogen acts on transmitter mechanisms in the brain, increasing
monoamine and neuropeptide receptors and binding sites in areas that control
emotion, behavior, and cognition. It helps to preserve and regenerate neuronal
elements, protects against inflammatory reactions and apoptosis, and contributes
anti-oxidant activity within the brain, partially by increasing brain magnesium.
Estrogen deficiency reduces cerebral blood flow (CBF), which is corrected by ERT,
and by physiologic levels of progesterone, which enhance cerebral artery
magnesium uptake. Poor CBF can decrease clearance of beta-amyloid, a pathogenic
factor in Alzheimer’s disease. Magnesium’s anticonstrictive effect is mediated by
several mechanisms. It is a natural calcium blocker, it increases prostacyclin
secretion, and it modulates nitric oxide synthesis and release. It also has
neuroprotective effects that have improved recovery in brain-traumatized animals
and that are being applied in stroke victims. Brain trauma or ischemia lower
magnesium brain levels, resulting in malfunction of key enzymes involved in energy
and nucleic acid metabolism, and in protein synthesis. Magnesium stabilizes
phospholipid components of cell membranes, protects against cellular disruption, and
counteracts initiation of secondary neurochemical changes and altered
neurotransmitter release, and free radical production.

3
The influence of some antipsychotic drugs on erythrocyte and plasmatic
magnesium level and of other bivalent cations
M. Nechifor 1 , C. Vaideanu 1 , I. Palamaru 2 , C. Borza 3 , I. Mindreci 4 , 1.Department of
Pharmacology; 2.Institute of Hygiene Public Health; 3.Department of Psychiatry; 4.
Department of Biophysics, Iasi, Romania. nechifor@umfiasi.ro
The aim of the study was to investigate the influence of haloperidol (4-10 mg/day)
and risperidone (Rispolept) (4-6 mg/day) on the plasmatic concentrations of
magnesium, calcium, copper and zinc, and on the erythrocyte magnesium level. We
determined the cations levels before the hospitalized adult patients received any
therapy and after 3 weeks of therapy. Our data indicate a lower erythrocyte
magnesium in schizophrenics before treatment compared to the level found in
controls (4,82±0,02 mg/dl in schizophrenic patients before treatment versus
5,82±0,11 mg/dl in control group, p

4
Effects of magnesium on blood-brain barrier breakdown and brain edema in
septic rats
Tulin Erdem M.D., Damla Aktan M.D., Ayse Erturer M.D., Mukadder Orhan M.D.,
Nahit Cakar M.D, Figen Esen M.D. University Of Istanbul Medical Faculty,
Department Of Anesthesiology And Intensive, Care Istanbul, Turkey.
Erdentn@yahoo.com
INTRODUCTION: Encephalopathy is a common complication of sepsis whose
severity correlates with mortality. Its now accepted that changes in the blood-brain
barrier (BBB) permeability plays a critical role in the pathophysiology of septic
encephalopathy. In this study, we determined the BBB permeability and brain edema
formation after abdominal sepsis produced by intraperitoneal implantation of a fibrin
thrombin cloth containing Escherichia coli, and effects of magnesium administration
on blood-brain barrier (BBB) and brain edema in septic rats.
METHODS: Experimental sepsis was induced in Sprague-Dawley rats by fibrinthrombin
cloth infected with 1,8x109 cfu/ml by McFarland standart 6 E.coli, injected
into the peritoneal cavity. The animals were randomly assigned to receive an
intramuscular bolus of either 270 mg/kg magnesium sulphate (G-5, n=22) or equal
volume of saline (G-2, n=22) 30 minutes after the induction of sepsis or 270 mg/kg
magnesium sulphate (G-4, n=23) 30 minutes before the induction of sepsis.
Nonseptic animals received saline (G-1, n=15) or magnesium sulphate (G-3, n=14)
with an identical protocol. BBB permeability was evaluated quantitatively 24 hours
after the induction of sepsis by spectrophotometric assay of Evans Blue dye (EBD)
extravasations. Specific gravity as an indicator of brain edema was measured 24
hours after the induction of sepsis.
RESULTS: Extravasation of EBD was evident in both hemispheres in three sepsis
groups (in group G-2, in group G-4, and in group G-5). Evans Blue dye content of
both hemispheres revealed a significant decrease in magnesium treated sepsis
group compared to saline treated sepsis group (in group G-4, and in group G-5)
(p0,05).
CONCLUSION: The present results indicate that BBB permeability defect occurs
after an intraperitoneal infected fibrin-thrombin cloth delivered model of sepsis and
magnesium seems to attenuate this defect

5
Magnesium-Vit B6 intake reduces central nervous hyperexcitability in children
*Mousain-Bosc M., **Roche M., °Rapin J.R., **Bali J.P. Departments of *Pediatry
and
**Biochemistry, CHU Nimes, 30029 NIMES CEDEX, France, °Department of
Pharmacology, Faculty of Medecine, University of Bourgogne, DIJON, France.
jean.pierre.bali@chu-nimes.fr
It has long been shown that Mg2+ depletion in mice causes hyperexcitability with
convulsive seizures; these effects were reversed by treatment with association of
magnesium-vit B6. Besides metabolic disorders, some genetic alterations could be
suspected in this pathology in which intracellular Mg2+ transport and storage may be
reduced without change in serum Mg2+ concentrations. In order to evaluate the
incidence of Mg2+/vitamin B6 regimen in the behaviour of hyperexcitable children,
we investigated on 52 children [0-15 years] and their family. To assess intracellular
Mg2+, we measured intra-erythrocyte Mg2+ levels (ERC-Mg). In our hands,
reference values for normal subjects were in the range [2.46 - 2.72 mmol/L]. In these
children, ERC-Mg values were lowered in 30/52 cases (2.041+/-0.279 mmol/L) and
Mg2+/vit B6 treatment (100 mg/day) for some weeks (3 to 24) restored normal ERC-
Mg values (2.329+/-0.386 mmol/L). In all patients, symptoms of hyperexcitability
(physical aggressivity, instability, scholar attention, hypertony, spasm, myoclony)
were reduced after 1 to 6 months of treatment. In addition, it must be noted that other
members of the same family shared similar symptoms with low ERC-Mg values and
Mg2+/vit B6 treatment also improved their clinical state. The case of two typical
families will be described. In conclusion, it can be postulated from this open study
that hyperexcitable children showed low ERC-Mg with normal serum Mg2+ values;
Mg2+/vit B6 intake restores normal ERC-Mg levels and improves the abnormal
behaviour of these children.

6
Effect of magnesium on neural activities in rat cortex and hippocampus in vitro
Keiichi Torimitsu, Nahoko Kasai, Yasuhiko Jimbo, Yuriko Furukawa, NTT Basic
Research Laboratories, Atsugi, Kanagawa, Japan. torimitu@will.brl.ntt.co.jp
Effect of magnesium on neural activities was investigated in rat cortex and
hippocampus in vitro. Measurements have been carried out for dissociated cell
cultures and slice preparations under low magnesium conditions. Electrical bursts,
calcium transients and glutamate release were investigated. 64-channel ITO
microelectrode array (electrode size: 10-20 µm square) and fluorescent
potentiometry with flow-cytometer were used for electrical measurement. Fluo-4 was
used for intracellular calcium measurement with two photon laser microscope.
Transient changes of glutamate release were investigated using an enzyme-based
electrochemical detection method. The method includes glutamate oxidase,
horseradish peroxidase and electron transfer mediator with the microelectrode array
described above. Embryonic cortical neurons (E18) were dissociated with papain and
cultured for 2-3 weeks in the medium containing DMEM and heat-inactivated serum.
Hippocampal slices were obtained from a P8 Wistar rat and cultivated on a porous
membrane for 7days. Spontaneous activities of the neurons were activated by a low
magnesium medium (LMGM). The LMGM induced a transient increase in glutamate
release and frequency modulation of intracellular Ca transients. Spatial distribution of
their activities was measured and imaged. In hippocampal slices, the responses
were differed among the area. Synaptic activities seem to be modulated by the
extracellular magnesium concentrations. Role of NMDA receptor and the mechanism
of magnesium modulation of synaptic activity have been considered. ATP
involvement in the synaptic modulation will also be discussed.

7
Magnesium gluconate offers no more protection than magnesium sulphate
following diffuse traumatic brain injury in rats
Renee J. Turner, Katherine W. DaSilva, Christine O’Connor, Corinna van den Heuvel
and Robert Vink. Department of Pathology, University of Adelaide, Adelaide, SA,
Australia Robert.Vink@adelaide.edu.au
Previous studies have demonstrated that magnesium salts, including the sulphate
and chloride forms, are neuroprotective following traumatic brain injury (TBI) 1 .
Recently, studies in cardiac ischaemia/reperfusion injury have demonstrated that the
gluconate salt of magnesium may provide superior protection against oxidative
damage and postischaemic dysfunction than MgSO4 2 . We have therefore compared
the efficacy of both MgSO4 and magnesium gluconate (MgGl2) on outcome following
diffuse TBI in rats. Adult male Sprague-Dawley rats were injured using the 2-metre
impact acceleration model of diffuse TBI as previously described 1 . At 30 min after
injury, animals were administered with either 250µmoles/kg i.v. MgSO4, MgGl2, or
equal volume saline vehicle. Thereafter, animals were assessed for motor and
cognitive outcome using the rotarod and Barnes maze, respectively. Treatment with
either magnesium salt significantly improved functional outcome as compared to
vehicle treated controls. There were no significant differences between the
magnesium treated groups. We conclude that MgSO4 and MgGl2 are equally
neuroprotective following TBI. Our results suggest that MgGl2 may only be more
effective in conditions that produce ischaemia, where high concentrations of reactive
oxygen species are generated.
1. Heath, D.L. and Vink, R. (1999) J. Pharmacol. Exp. Ther. 288: 1311-1316.
2. Murthi, S.B., Wise, R,M., Weglicki, W.B., Komarov, A.M. and Kramer, J.H.
(2003) Mol. Cell. Biochem. 45: 141-148.

8
Magnesium transport genes in yeast and plants
Richard Gardner, Jong-min Lee, Keith Richards, Salam Salih, Paul Donaldson*, Van
Kelly, Revel Drummond, 'Ana Tutone, School of Biological Sciences (or *Faculty of
Medical and Health Sciences), University of Auckland, Private Bag 92019, Auckland,
New Zealand. r.gardner@auckland.ac.nz
The magnesium uptake system in yeast consists of two homologous genes, ALR1
and ALR2 (named because overexpression confers aluminium resistance). The ALR
genes encode proteins homologous to the bacterial CorA family of magnesium
transporters. Mutagenesis of ALR1 has shown that the region of the protein
containing the three putative transmembrane domains is essential for its function, in
accord with results from the bacterial CorA gene. Initial electrophysiological analysis
of ALR1 suggests that it acts as a channel and that it mediates both inward and
outward movement of magnesium ions. Mg-dependent inward currents are transient
and operate only at high negative membrane potential. We previously showed that
aluminium ions (Al3+) block uptake of cations by ALR1, and suggested that this may
be the mechanism by which this environmental toxin inhibits growth of yeast cells.
Comparison of aluminium treatment and magnesium starvation of yeast has provided
experimental support for this idea. Recently a gene family of ten CorA homologues
have been identified in the higher plant Arabidopsis thaliana. Initial evidence
suggests that the different proteins may be localised to different cellular
compartments.

9
Molecular biology of magnesium transport proteins
Matin Piskacek*, Jochen Stadler*, Martin Kolisek*, Gabor Zsurka*, Julian
Weghuber*, Monika Schweigel**, Rudolf J. Schweyen*. * Vienna Biocenter, Institute
of Microbiology and Genetics, University of Vienna, Dr. Bohrgasse, A-1030 Vienna,
Austria; ** Free University Berlin, Institute of Veterinary-Physiology, Oertzenweg 19b,
D-14163 Berlin, Germany. rudolf.schweyen@univie.ac.at
We have recently characterized a mitochondrial protein (named Mrs2p) as
constituting the major mitochondrial Mg2+ transport protein in the yeast
Saccharomyces cerevisiae. Combined physiology and molecular studies suggest
that it forms a channel in the inner mitochondrial membrane mediating high capacity
Mg2+ influx, driven by the inside negative membrane potential. This protein is a
distant homologue of the bacterial CorA Mg2+ transport protein and it has well
conserved homologues in all eukaryotes. The human genome encodes a single Mrs2
homologue. Work is in progress to characterize its role in Mg2+ influx into
mitochondrial DNA by using cell cultures overexpressing hsMrs2p and cell cultures
with knock-down of its mRNA.

10
Mg2+ transport proteins of the yeast plasma membrane and cytoplasmic
homeostasis factors
Jochen Stadler, Anton Graschopf, Stefan Köstler, Rudolf J, Schweyen. Max Perutz
Laboratories, Department of Microbiology and Genetics / University of Vienna,
Campus Vienna Biocenter Dr. Bohrgasse 9, A-1030 Vienna, Austria.
rudolf.schweyen@univie.ac.at
The major Mg2+ influx into yeast cells is mediated by Alr1p, an integral protein of the
plasma membrane. Alr1p is a distant relative of the bacterial CorAp and
mitochondrial Mrs2p Mg2+ transport proteins. Like these proteins, Alr1p appears to
form oligomeric complexes in the plasma membrane, which probably constitute
Mg2+ channels. Regulation of Mg2+ fluxes into the cell is mediated by transcriptional
control of the ALR1 gene and, more importantly, by Alr1p endocytosis and
degradation in the vacuole. A novel factor, Cos16p, has been identified as being an
integral protein of the endoplasmic reticulum (ER) and essential for tolerance of
yeast cells to elevated extracellular Mg2+ concentrations. Unlike Alr1p, which occurs
in lower eukaryotes only, Cos16p is well conserved from yeast to man. Its
overexpression or absence in yeast cells causes a decrease or increase,
respectively of total cellular Mg2+ concentrations. We speculate on the involvement
of Cos16p in the Mg2+ exchange between the endoplasmic reticulum and the
cytoplasm. As revealed from whole genome expression profiling, Mg2+ starvation of
yeast cells elicits a highly significant upshift of gene activities by the calcineurin
pathway, similar to Ca2+ overload. Work is in progress to learn which regulatory
mechanisms are involved in this response.

11
The pathogenesis of Machado-Josephs Disease; a high manganese / low
magnesium induced CAG expansion mutation in susceptible genotypes?
Mark Purdey. High Barn Farm, Elworthy, Taunton, Somerset, Ta43px, UK.
Tsepurdey@aol.com
The origins of the progressive spinocerebellar ataxic disorder ‘Machado Josephs
Disease’ (MJD) has been solely attributed to an autosomal dominant inheritance of
an expansion mutation in the CAG repeat in chromosome 14q32.1; a mutation that
has purportedly been disseminated widely since the Middle Ages into Azorean,
Dutch and Makassan communities by an international trading community of
Sephardic Jews based in NE-central Portugal. However, the majority of the families
implicated in the increasing number of familial MJD clusters being observed around
the world - in Aboriginal, African, Asian and Yemeni families - cannot be connected
back to the Portuguese originators. An analytical study of the isolated ecosystems
supporting both the Portuguese and non-Portuguese MJD affected communities
demonstrate a common abnormal hallmark of high manganese / low magnesium
status; suggesting that this aberrant mineral ratio inactivates the Mn / Mg catalysed
endonuclease 1 enzyme in the biosystems of those residing in these ecosystems.
Endonuclease activity is crucial for protecting against the expansion / contraction of
the trinucleotide repeats in the genes that encode for proteins such as Ataxin 3 - the
mutant protein that is central to the pathogenesis of MJD. It is proposed that MJD,
and, possibly the other more common expansion mutation diseases such as
Friedrich's Ataxia / Huntingdon's Chorea, are multifactorial diseases caused by a
hitherto unrecognised autosomal dominant inherited failure to regulate Mn / Mg
metabolism in populations dependent upon high Mn / Low Mg ecosystems. Mg
supplementation of the 'at risk' populations during the 'in utero' developmental stages
could be all that is required to maintain healthy endonuclease turn over, thereby
protecting MJD susceptible genotypes against this fatal, progressive
neurodegenerative disease.

12
Interest of stable isotopes use in the evaluation of Mg metabolism
Charles Coudray, Christine Feillet-Coudray, André Mazur and Yves Rayssiguier,
INRA, Clermont-Ferrand/Theix France. coudray@clermont.inra.fr
Magnesium (Mg) is a biologically essential mineral and Mg deficiency is known to
lead to severe biochemical and symptomatic disorders. Radioactive isotopes and,
more recently stable isotopes, have been used as research tools to determine
intestinal Mg absorption in man and animal under different nutritional and
physiological conditions. Mg isotopes are given orally or orally plus intravenously and
determined in feces and/or in plasma and urine to calculate intestinal Mg absorption
and possibly endogenous Mg excretion. Mg isotopes have been used to assess Mg
exchangeable pools under nutritional and physio-pathological conditions. Mg
isotopes are given intravenously and determined in plasma and urine to calculate the
size and the half-life of the different Mg exchangeable pools. Mg isotopes have been
also used to study the mechanisms of Mg cellular exchanges. For that, erythrocytes
or other types of cells is loaded with Mg isotope or incubated in isotope-rich medium
to study the flux of Mg and its mechanisms. More recently, new in vitro tests have
been developed to study the avidity of cells for Mg in different nutritional and genetic
conditions. Whole blood is incubated with Mg isotopes and isotopic blood cell
enrichment is measured which reflects the need of cell to Mg and thus its initial
status. The present paper is a report concerning the use of Mg stable isotopes and
their advantages in these different fields of Mg absorption and metabolism. Available
studies demonstrated clearly that stable isotopes are a useful research tool for
determining intestinal Mg absorption and a precious research tool for Mg cellular
exchanges and exchangeable Mg pools exploration to evaluate Mg status.

13
Effects of intracellular concentrations of Na+ and Mg2+ on Mg2+ transport in
rat ventricular myocytes
Masato Konishi, Michiko Tashiro and Pulat Tursun. Department of Physiology, Tokyo
Medical University, Shinjuku-ku Tokyo, Japan. mkonishi@tokyo-med.ac.jp
To study cellular Mg2+ transport, cytoplasmic Mg2+ concentration ([Mg2+]i) of rat
ventricular myocytes was measured with a fluorescent indicator furaptra (mag-fura-2)
under Ca2+-free conditions at 25°C. After [Mg2+]i was increased in the solution
containing 24-93 mM Mg2+, the reduction of extracellular Mg2+ to 1 mM caused a
rapid decrease in [Mg2+]i only in the presence of extracellular Na+ (extracellular
Na+-dependent Mg2+ efflux). The rate of decrease in [Mg2+]i (d[Mg2+]i) was
significantly greater at the higher initial levels of [Mg2+]i, and was nearly zero at the
basal levels; the efflux was half activated at 1.4 mM [Mg2+]i with a maximum
d[Mg2+]i of 0.0022 mM/s. We then studied intracellular Na+-dependence of the Mg
efflux by comparing d[Mg2+]i under different levels of intracellular Na+ concentration
([Na+]i). After 1 mM ouabain treatment in the solution containing 106 mM Na+ for 3
hours, average [Na+]i estimated with a fluorescent indicator SBFI was increased
from 12 mM to 48 mM (Na+-loaded cells). On the other hand, low extracellular Na+
concentration (1.3 mM) caused a decrease in [Na+]i to about 0 mM in 3 hours (Na+depleted
cells). The average d[Mg2+]i was significantly smaller in the Na+-loaded
cells. In some experiments, the myocytes were voltage clamped at -80 mV using the
perforated patch-clamp technique with amphotericin B. We found that the increase
in pipette Na+ concentration from 0 mM to 130 mM significantly decreased d[Mg2+]i.
The results are consistent with the Mg2+ efflux activity driven by the [Na+] gradient
across cell membrane, or the Na+-Mg2+ exchange.

14
Depletion of intracellular Mg2+ via Na+-independent passive Mg2+ pathways
Shinsuke Nakayama, Hideki Nomura, Lorraine M. Smith, Joseph F. Clark*, Tadayuki
Uetani# & Tatsuaki Matsubara, #Department of Cell Physiology, and #Department of
Metabolic Diseases, Nagoya University Graduate School of Medicine, Aichi, Nagoya
466-8550, Japan, and *Department of Neurology, University of Cincinnati Medical
Center, Cincinnati OH 45267-0525, USA. h44673a@nucc.cc.nagoya-u.ac.jp
Epidemiological studies suggest that magnesium deficiency is correlated to many
important diseases, e.g. diabetes mellitus, hypertension, cardiovascular and
cerebrovascular diseases. We employed 31P-nuclear magnetic resonance in the
guinea-pig taenia caeci, which is a smooth-muscle-rich tissue and can be used a
good model for studying regulatory mechanisms for the intracellular free Mg2+
concentration ([Mg2+]i). [Mg2+]i was estimated from the chemical shifts of the ATP
peaks. Simultaneous removal of Mg2+ and Ca2+ from the extracellular medium,
rapidly decreased [Mg2+]i (by ~30-fold after 100 min) even when extracellular Na+
was replaced with K+. This suggests the presence of Na+-independent Mg2+
pathways (i.e. Mg2+ pathways other than Na+-Mg2+ exchange) blocked by divalent
cations. In contrast to K+ substitution, when extracellular Na+ was substituted with
large monovalent cations, such as NMDG (N-methyl-D-glucamine) or choline,
simultaneous removal of divalent cations decreased [Mg2+]i only very slowly. In Li+
substitution for Na+, changes in the chemical shifts of the ATP peaks were also
small. Application of calibration curves for a Li+-rich solution however reveled a
significant [Mg2+]i fall comparable to that in the K+ substitution. The properties of
LTRPC7, newly identified as a Mg2+- and Ca2+-permeable divalent cation channel,
can account for the depletion of intracellular Mg2+ seen in this study under Na+-free
conditions.

15
Set point shift of the intracellular Mg2+ concentration with amiloride and KB-
R7943 in vascular smooth muscle
Uetani Tadayukik, Yukihisa Hamaguchi, Yasushi Tatematsu, Shinsuke Nakayama.
Department of Cell Physiology, Nagoya University Graduate School of Medicine,
Nagoya, Japan. h44673a@nucc.cc.nagoya-u.ac.jp
It has been suggested that intracellular free magnesium concentration ([Mg2+]i) is
regulated by Na+-Mg2+ exchange. It is interesting to examine the effect of amiloride
which inhibits many Na+-dependent transporters, in terms of [Mg2+]i regulation in
vascular smooth muscle. We measured the intracellular free Mg2+ concentration
([Mg2+]i) using 31P NMR in pig carotid artery smooth muscle. In normal solution,
application of amiloride (1 mM) decreased [Mg2+]i by~12% after 100 min.
Subsequent washout tended to further decrease [Mg2+]i. In contrast, application of
amiloride significantly increased [Mg2+]i (by~13% after 100 min) under Ca2+-free
conditions, where passive Mg2+ influx is facilitated. The treatments had little effect
on intracellular ATP and pH (pHi). Essentially the same results were reproduced with
KB-R7943, a selective blocker of reverse mode Na+-Ca2+ exchange. Application of
dimethyl amiloride (0.1 mM) in the presence of Ca2+ hardly changed [Mg2+]i,
although it inhibited Na+-H+ exchange at the same concentration. Removal of
extracellular Na+ caused a marginal increase in [Mg2+]i after 100~200 min, as seen
in intestinal smooth muscle in which Na+-Mg2+ exchange is known to be the primary
mechanism of maintaining a low [Mg2+]i against electro-chemical equilibrium. In
Na+-free solution (containing Ca2+), amiloride did not increase, but rather increased
[Mg2+]i. The results suggest that Na+-Mg2+ exchange play a crucial role in
determining [Mg2+]i in vascular smooth muscle cells. We supposed that the
decrease in [Mg2+]I caused by amiloride is associated with inhibition of Mg2+
uptake. It has been suggested that KB-R7943 inhibits not only Na+/Ca2+ exchange
but also Na+/ Mg2+ exchange.

16
Magnesium and the inflammatory response: potential physiological and
clinical implications
Rayssiguier Yves, Mazur André. INRA, UMMM Theix, St Genes Champanelle,
France. yves.rayssiguier@clermont.inra.fr
Inflammation is a biological response of the immune system to a variety of
aggressions but a damaging effect occurs when the initial response becomes
amplified and then dysregulated. Excessive response may be, for instance, involved
in diseases such as atherosclerosis, diabetes, cerebral and myocardial ischemia,
septic shock syndrome, severe trauma or skeletal fragility. Experimental magnesium
deficiency in the rat induces after few days a clinically inflammatory syndrome
characterized by PMN and macrophage activation, release of inflammatory cytokines
and acute phase proteins, and excessive production of free radicals. Mg-deficient
females have a greater degree of protection against inflammation and oxidative
damage than Mg-deficient males suggesting that different physiological background
created by the female sex steroid hormones may offer some protection against these
conditions. The effect of various Mg concentrations on ROS production was
investigated in vitro. Increasing concentration of Mg was shown to decrease PMN
respiratory burst. Similar observations were made using rat and human PMN.
Moreover, in vitro cytokine production by whole blood from human volunteers was
decreased by high Mg concentration. Thus, increasing external Mg concentration
decreases inflammatory response while a reduction in the extracellular Mg
concentration results in cell activation. Because Mg acts as a natural Ca antagonist,
the molecular basis for inflammatory response in Mg deficiency is probably the result
of modulation of intracellular Ca concentration. This review will summarize recent
findings supporting the role of inflammatory processes in Mg deficiency and will
discuss why Mg is therapeutically effective.

17
Magnesium Sulfate- a role in the management of Irukandji Syndrome?
Michael Corkeron, Intensive Care Unit, The Townsville Hospital, Townsville, Qld,
4814 Australia. Michael_Corkeron@health.qld.gov.au
Irukandji Syndrome is characterised by pain, a hyperadrenergic state and not
infrequently, markers of myocardial injury. It affects approximately 100 individuals per
year in Northern Australia and its severity ranges from troublesome to lifethreatening;
two individuals are known to have died from complications of severe
hypertension. Treatment is symptomatic, and often less than satisfactory for victims
and their carers.
The culprit organisms are known to be carybdeid jellyfish; the best known of these is
Carukia barnesi, though several species are thought to be implicated. The
mechanisms of venom action in irukandji syndrome are incompletely understood and
there remains considerable debate as to these. In a piglet model noradrenaline levels
increase several hundredfold after experimental envenoming by C. barnesi tentacle
extract.
Magnesium sulfate has been used in a number of patients with Irukandji Syndrome
during 2003, either as monotherapy or in conjunction with other agents. Experience
with this treatment and future research directions in this area are described.

18
The effect of magnesium deficiency on primary tumour growth and the
effectiveness of antitumour treatment in transplantable murine cancers
Anna Nasulewicz 1,2 , Joanna Wietrzyk 1 , Yves Rayssiguier 2 , André Mazur 2 , Adam
Opolski 1 , 1 Institute of Immunology and Experimental Therapy, Polish Academy of
Sciences, 12, R. Weigla St., 53-114 Wroclaw, Poland, 2 CRNH d'Auvergne, Unité
Maladies Métaboliques et Micronutriments, INRA, Theix, 63122 St Genès
Champanelle, France. nasulewicz@iitd.pan.wroc.pl
The aim of this work was to examine the relationship between Mg status and tumour
growth in mice. The results show a significant retardation of primary tumour growth in
mice receiving Mg-deficient diet by: 60% for Lewis lung carcinoma (LLC), 57% for
colon adenocarcinoma (C38), 75% for mammary carcinoma (16/C) and 30% for
melanoma (B16) comparing to the control diet fed group. Mg depletion followed by
repletion leads to the significant increase of primary LLC tumour burden (142% of the
control tumour weight). Even if long-term Mg deficiency results in marked decrease
of Mg concentration in plasma, Mg concentration in tumour cells is weakly affected.
Mg depletion leads to changes in gene expression in LLC cancer cells. The study of
differential gene expression by cDNA array has shown that several genes are
regulated by Mg deficiency (up-regulated: vitamin D 24-hydroxylase, glycoprotein V,
titin and gelsolin; down-regulated: APC, fyn, JNK 3, metalloproteinase 9, laminin
subunit, integrin and FGFR1). Moreover, we have shown that Mg depletion enhances
the efficacy of antitumour treatment. The life span of P388 leukaemia-bearing mice,
kept on Mg-deficient diet and treated with doxorubicin, was higher then those of
identically treated animals, which were fed control diet (133% vs. 99% as compared
to the control untreated group kept on Mg-sufficient diet).

19
Investigation of intracellular magnesium mobilization pathways in PC12 cells
by simultaneous Mg-Ca fluorescent imaging
Takeshi Kubota 1 , Hirokazu Komatsu 4 , Yutaka Shindo 2 , Kentaro Tokuno 2 , Yoshiichiro
Kitamura 1 , Hiroto Ogawa 3 , Koji Suzuki 4 , and Kotaro Oka 1 , 1 Center for Life Science
and Technology, Keio University, Yokohama, Japan, 2 Department of System Design
Engineering, Keio University, Yokohama, Japan, 3 Department of Biology, Saitama
Medical School, Saitama, Japan, 4 Department of Applied Chemistry, Keio University,
Yokohama, Japan. kubota@bpni.bio.keio.ac.jp
Intracellular Mg2+ concentration shows dynamic change in response to various
extracellular stimuli in excitable cells, and it suggests that the concentration change
plays important roles in intracellular signal transduction. To clarify the role of
intracellular Mg2+ and Ca2+ during stimulation, we examined the correlation of these
divalent cations in PC12 cells by fluorescent imaging techniques. At first, we
developed a novel fluorescent Mg indicator KMG-104 (Ex = 504 nm, Em = 523 nm).
The dissociation constant (Kd) of KMG-104 is 3 mM, near to the intracellular Mg2+
concentration (about 1 mM), and not affected by Ca2+ disturbance. PC12 cells were
double-stained by KMG-104 and a Ca2+ indicator, Fura-Red, and the fluorescences
exited at 480 nm from these dyes were acquired simultaneously with a W-View
optical system with a 535/55 BP (KMG-104) and a 600 LP (Fura-Red) filters,
respectively. When the cells were stimulated with a mitochondrial uncoupler, FCCP
(3 µM), both Mg2+ and Ca2+ increased. After washing the FCCP, Mg2+
concentration returned to the basal level, but Ca2+ concentration did not recover. To
investigate the correlation of Ca2+ and Mg2+ mobilization directly, intracellular Ca2+
concentration was increased by photolysis of caged-Ca2+. Transient increase of
Ca2+ caused no Mg2+ concentration change. From these results, we presume Mg2+
mobilization pathway does not correspond with Ca2+ cascades. A plausible source
of Mg2+ mobilization is Mg2+ release from intracellular Mg-ATP complex by ATP
consumptions.

20
Magnesium attenuates post-traumatic depression/anxiety following diffuse
traumatic brain injury in rats
Lisa Fromm 1 , Deanne L. Heath 1 , Robert Vink 2 and Alan J. Nimmo 1 . 1 School of
Pharmacy and Molecular Sciences, James Cook University, Townsville, QLD;
2 Department of Pathology, University of Adelaide, Adelaide SA, AUSTRALIA.
Alan.Nimmo@jcu.edu.au
Magnesium has been shown to decline after traumatic brain injury (TBI), and this
decline is thought to be associated with the ensuing neuronal cell death and
subsequent functional impairment. While the effects of magnesium on motor and
cognitive deficits have been well studied following TBI, few studies have addressed
post-traumatic depression as an outcome parameter, despite it being a major clinical
problem with an incidence of between 6 and 77%. This study investigated the
incidence of post-traumatic depression/anxiety in an animal model of diffuse TBI, and
also explored the use of magnesium sulphate as an interventional treatment. Diffuse
TBI was induced in anaesthetised, adult, male Sprague-Dawley rats (n=32) using the
2 m impact-acceleration model of injury. At 30 min after injury, half of the animals
received 250µmol/kg i.v. MgSO4 while the other half served as non-treated controls.
Before and for 6 weeks after injury, the open-field, spontaneous activity test was
used to determine post-traumatic depression/anxiety relative to pre-injury. In this
test, animals are placed in a 1 metre square box with 100 squares marked on the
base. The number of squares entered in a 5-min period is recorded. Incidence of
post-traumatic depression/anxiety was defined as the number of animals
demonstrating a reduction in spontaneous activity to less than 100 squares in 5 min.
Prior to injury, rats typically entered a mean of 201 ± 12 (SEM) squares over a 5 min
observation period. At 1 week after injury, non-treated animals had a mean score of
62 ± 13. The incidence of post-traumatic depression/anxiety in these animals was
61%, which is similar to that observed clinically. In contrast, animals treated with
MgSO4 had a mean activity score of 144 ± 23 at 1 week after TBI and an incidence of
depression/anxiety of less that 30%. The significant difference between groups
persisted for the entire 6-week observation period. The improvement in posttraumatic
depression/anxiety conferred by magnesium adds further weight to
available evidence of magnesium’s benefit as a neuroprotective agent after TBI.

21
The CorA-Mrs2 superfamily of Mg2+ transport proteins in bacteria and
mitochondria
Elisabeth Froschauer 1 , Martin Kolisek 1 , Martin Piskacek 1 , Gabor Zsurka 1 , Julian
Weghuber 1 , Monika Schweigel 2 , Rudolf J. Schweyen 1 . 1 Max Perutz Laboratories,
Department of Microbiology and Genetics / University of Vienna, Campus Vienna
Biocenter Dr. Bohrgasse 9, A-1030 Vienna, Austria; 2 Free University Berlin,
Institute of Veterinary-Physiology, Oertzenweg 19b, D-14163 Berlin, Germany.
rudolf.schweyen@univie.ac.at
The CorA proteins of bacteria and the Mrs2 proteins of mitochondria are
characterized by the presence of two adjacent transmembrane domains. The only
sequence conserved among them (Y/F-G-M-N) is a short motif at the end of one of
these domains. Functional homology of these proteins is evident from the fact that
mitochondrial defects of Mrs2p lacking yeast cells (mrs2D mutants) are cured by the
expression of either the bacterial CorA protein or the human Mrs2 protein. Growth of
CorA mutant bacterial cells are Mg2+ -dependent, mrs2D yeast cells show
unconditional mitochondrial defects, but stay viable, while human mrs2D cells are
unviable.
Similarity between CorA and Mrs2 proteins further extends to the Mg2+ fluxes across
membranes. By use of the fluorescent, Mg2+ -sensitive dye mag-fura 2 we have
recently determined Mg2+ fluxes across bacterial plasma membranes as well as
across yeast and human mitochondrial membranes. The CorA and Mrs2 proteins
mediate rapid, high- capacity influx into bacterial cells and into isolated mitochondria,
respectively. Addition of 10 mM Mg2+ to isolated mitochondria can result in an
increase of intramitochondrial free Mg2+ by as much as 50% per second, with a
pronounced saturation at about 5 mM in yeast mitochondria. The driving force for
Mg2+ influx by both, CorA and Mrs2 proteins is the high (inside negative) membrane
potential of mitochondria and bacteria. Mg2+ influx studies suggest the presence of
Mg2+ channels in the bacterial and mitochondrial membranes. This notion is
supported by cross-linking studies in yeast and human mitochondrial membranes,
revealing the presence of homo-oligomeric Mrs2 protein complexes.

22
Effects of Oral Magnesium Therapy on Exercise Tolerance, Exercise-Induced
Chest Pain, and Quality of Life in Patients with Coronary Artery Disease
Michael Shechter, MD, MA 1,2, C. Noel Bairey Merz, MD 2, Hermann-Georg
Stuehlinger, MD 3, Joerg Slany, MD 4, Otmar Pachinger, MD 5, Burton Silver,
PhD6, and Babeth Rabinowitz, MD 1 From 1 The Heart Institute, Sheba Medical
Center, Tel Hashomer and Tel Aviv University, Israel, 2 Preventive & Rehabilitative
Cardiac Center, Division of Cardiology, Department of Medicine, Cedars-Sinai
Medical Center and the UCLA School of Medicine, Los Angeles, California, USA, 3
Department of Emergency Medicine, General Hospital, Vienna, 4 2nd Department
of Medicine, KA Rudolfstiftung, Vienna, and 5 Department of Cardiology, University
Hospital, Innsbruck, Austria, 6 IntraCellular Diagnostics, Inc., Foster City,
California, USA. shechtes@netvision.net.il
Previous work has demonstrated that magnesium supplementation improves
endothelial function in patients with coronary artery disease (CAD). However the
impact on clinical outcomes, such as exercise-induced chest pain, exercise
tolerance and quality of life, has not been established. In a multi-center, multinational,
prospective, randomized, double-blind and placebo-controlled trial, 187
CAD patients (151 men, 36 women, mean ± SD age 63 ± 10 years, age range 42
to 83 years), were randomized to receive either oral magnesium 15 mmol twice
daily (Magnosolv Granulat, total magnesium 365 mg provided as magnesium
citrate, Asta Medica, Vienna, Austria) (n=94) or placebo (n=93) for 6 months.
Symptom-limited exercise testing (Bruce protocol) and quality of life questionnaires
were the outcomes measured. Intracellular magnesium levels ([Mg]i) were
assessed in a sub-study of 106 consecutive patients from sublingual cells by x-ray
dispersion (EXATM) (normal mean ± SD values 37.9 ± 4.0 mEq/L). Magnesium
therapy significantly increased [Mg]i in the sub-study at 6 months compared to
placebo (35.5 ± 3.7 versus 32.6 ± 2.9 mEq/L, p=0.0151). Magnesium treatment
significantly increased exercise duration time compared to placebo (8.7 ± 2.1
versus 7.8 ± 2.9 minutes, p=0.0075). At 6 months, exercise induced chest pain
was notably less in patients who had received magnesium compared to placebo
(8% versus 21%, p=0.0237), causing severe enough angina to terminate the test in
7% of those receiving magnesium, in contrast to 17% of placebo group. Quality of
life parameters had significantly improved in the magnesium group. These findings
suggest that oral magnesium supplementation in CAD patients for 6 months,
results in a significant improvement in exercise tolerance, exercise-induced chest
pain, and quality of life, suggesting a potential mechanism whereby magnesium
could beneficially alter outcomes in CAD patients.

23
A prospective non-randomized, open-labeled pilot study investigating the use
of magnesium in patients undergoing non-acute percutaneous coronary
intervention with stent implantation
Vladimir Rukshin, M.D., Raul Santos, M.D., Mitch Gheorghiu, M.D., Prediman K.
Shah, M.D., Saibal Kar, M.D., Sriram Padmanabhan, M.D., Babak Azarbal, M.D.,
Vivian T. Tsang, B.S., Raj Makkar, M.D., Bruce Samuels, M.D., Norman Lepor, M.D.,
Ivor Geft, M.D., Steve Tabak, M.D., Mehran Khorsandhi, M.D., Neil Buchbinder,
M.D., Neil Eigler, M.D., Bojan Cercek, M.D., Keta Hodgson, R.N., and Sanjay Kaul,
M.D.
From the Vascular Physiology and Thrombosis Research Laboratory of the
Atherosclerosis Research Center, the Burns and Allen Research Institute, the
Division of Cardiology and the Department of Medicine, Cedars-Sinai Medical Center
and UCLA School of Medicine, Los Angeles, California, USA. kaul@cshs.org
BACKGROUND - Magnesium has recently been shown to inhibit acute stent
thrombosis in animal models. This study tested the feasibility of magnesium
administration in patients undergoing non-acute percutaneous coronary intervention
(PCI) with stent implantation.
METHODS - Twenty-one patients undergoing non-emergent PCI were enrolled and
received intravenous MgSO4 (2g bolus over 20 minutes pre-PCI followed by 14g
over 12 hours infusion). Endpoints: safety outcomes - hypotension, bradycardia,
bleeding complications and heart block within first 24 hours; angiographic outcomes -
acute thrombotic occlusion and need for platelet GP IIb/IIIa inhibitor (GPI) bailout;
and clinical outcomes - death, myocardial infarction (MI), recurrent ischemia and
need for urgent revascularization at 48 hours and 30 days.
RESULTS - No significant effects on heart rate or blood pressure, major bleeding
complication or new ECG changes were observed. Angiographic thrombus was
visualized in 2 cases, and coronary artery dissection in 1 case post-stent
deployment. None of these cases required the use of GPI for bailout. Death, MI,
recurrent ischemia and need for urgent revascularization were not observed. Serum
magnesium level increased from 2.1+0.3 at baseline to 3.5+0.8 mg/dL at the end of
infusion (P

24
Optimal administration dosage of magnesium sulfate for torsades de pointes
in children with long QT syndrome
Kenji Hoshino, Kiyoshi Ogawa, Takashi Hishitani, Yoshikatsu Etoh*. Department of
Pediatric Cardiology, Saitama Children's Medical Center, Iwatsuki City, Saitama,
Japan, * Department of Pediatrics, the Jikei University School of Medicine.
a0182608@pref.saitama.jp
BACKGROUND: Administration of magnesium sulfate (MgSO4) is a very effective
and safe treatment for torsades de pointes (TdP) associated with acquired long QT
syndrome (LQTS) in adults. We discussed the efficacy of MgSO4 for TdP in children
with congenital and acquired LQTS. We also discussed the optimal administration
dosage and serum magnesium (SMg) concentration during MgSO4 therapy.
METHODS: We studied 7 consecutive LQTS children undergoing MgSO4 therapy for
TdP. Of the 7 children, five were congenital LQTS and two were acquired LQTS. A
bolus injection of MgSO4 was given intravenously until the TdP was completely
abolished or the maximum bolus dosage (12mg/kg) over 1 to 2 minutes. Continuous
infusion of MgSO4 was given over 1 to 7 days.
RESULTS: Of the 7 patients, six responded completely to the initial bolus. The bolus
dosage was 5.9±3.8mg/kg (range, 2.3-12mg/kg) in these six, and the other remaining
one (neonate with congenital LQTS) required a total of 30mg/kg until the complete
abolishment. The continuous infusion was given at rates of 0.3 to 1.0mg/kg/hr and
patients did not show recurrence of TdP. The SMg concentration was 3.9±0.9mg/dl
(2.9-5.4mg/dl) immediately after bolus injection. The mean corrected QT (QTc)
interval before and after bolus injection did not show significant difference.
CONCLUSIONS: Intravenous infusion of MgSO4 was effective for TdP in children
with LQTS, and MgSO4 abolished TdP without shortening QTc interval. The optimal
bolus dosage, infusion rates and SMg concentration were 3 to 12mg/kg, 0.5 to
1.0mg/kg/hr and 3 to 5mg/dl, respectively.

25
Effects of Mg2+ on cardiac excitation-contraction coupling
Michailova AP, Belik, ME, and McCulloch, AD. Department of Bioengineering,
University of California, San Diego, USA. amihaylo@bioeng.ucsd.edu
Magnesium regulates a large number of cellular processes. Small changes in
intracellular free Mg2+ ([Mg2+]i) may have important effects on cardiac excitability
and contractility. For this reason, we used our ionic-metabolic model (1) - that
incorporates equations for Ca2+ and Mg2+ buffering and transport by ATP and ADP
and equations for MgATP regulation of ion transporters (Na+-K+ pump, sarcolemmal
and sarcoplasmic Ca2+ pumps) – to investigate the effects of [Mg2+]i on cardiac
excitation-contraction coupling. Model results indicate that variations in cytosolic
Mg2+ level might sensitively affect diastolic and systolic Ca2+, sarcoplasmic Ca2+
content, Ca2+ influx through L-type channels, efficiency of the Na+/Ca2+ exchanger
and action potential shape. The analysis suggests that the most important reason for
the observed effects is a modified normal function of sarcoplasmic Ca2+-ATPase
pump by altered diastolic MgATP levels. The model is able to reproduce qualitatively
a sequence of events that correspond well with experimental observations during
cardiac excitation-contraction coupling in mammalian ventricular myocytes (2).
1. Michailova AP, and McCulloch AD. (2001) Model Study of ATP and ADP
Buffering, Transport of Ca2+ and Mg2+, and Regulation of Ion Pumps in
Ventricular Myocyte. Biophysical Journal 81:614-629.
2. Agus ZA, Kelepouris, E, Dukes I, and Morad M. (1989) Cytosolic magnesium
modulates calcium channel activity in mammalian ventricular cells. American
Journal Physiology 256:C425-C455.

26
Intracellular magnesium assay correlations with: oral & transdermal
absorption, endothelial function, exercise tolerance, and chronic disease
depletion.
Burton B. Silver#, C. Norman Shealy*, Michael Shechter*^, Mark C.P Haigney**,
Allison J. Stewart ##, Kim Brower*. ** The Department of Medicine, Division of
Cardiology, Uniformed Services University of the Health Sciences, 4301 Jones
Bridge Rd. Bethesda, MD, # IntraCellular Diagnostics, Inc. 553 Pilgrim Dr (B) Foster
City, CA, *^Division of Cardiology, Dept Medicine, Cedars-Sinai Medical Center, Los
Angeles, CA, * Holos Institutes of Health, Springfield, MO, ## Dept of Veterinary
Med., Ohio St U. Columbus. OH, USA. DocBurt@aol.com
Energy dispersive X-ray microanalysis (EXAtm) for non-invasive intracellular
measurement of magnesium has been proven to be a valuable tool in multiple
aspects of normal as well as pathological magnesium metabolism. Over the last 20
years data has accumulated using this unique method, which is now available to
accurately evaluate tissue Mg levels. While serum levels of Mg give little information
about a patients overall status with respect to this essential mineral, utilizing
intracellular analysis it has been determined that Mg levels are significantly reduced
in multiple patho-physiological states. Description of the methodology and
experimental data will focus on intracellular [Mg2+]i determinations obtained from
subjects repleted with Mg or with syndromes related to Mg deficiency. Intracellular
analysis studies (EXAtm) of tissue magnesium levels include:
• Effects of oral and transdermal Mg on tissue levels in CAD patients.
• Effects of intracellular Mg on exercise and endothelial function.
• Intracellular magnesium’s role in dysrhythmias, heart failure, and infarction
patients.
• Correlations of intracellular Mg with cardiac tissue.
• Depletion studies of intracellular Mg in human subjects and large animals.
RATIONALE: Magnesium’s role in intracellular metabolism is second only in
potassium as the most common intracellular cation in human physiology. Multiple
normal functions, as well as pathological syndromes, are affected by the cellular
levels and availability of the magnesium stores within tissues. Logically, measuring
magnesium accurately and easily within tissues is of immense use to the
understanding and treatment of a host of diseases in the human condition.
Applications of intracellular magnesium analysis will be reported from investigational
data incorporating this unique method.

27
Intracellular magnesium in furosemide treated patients with congestive heart
failure suffering from diabetes mellitus and/or renal dysfunction
I. Alon, D. Almoznino-Sarafian, S. Berman, O. Gorelik, M. Shteinshnaider, D.Modai,
N. Cohen, Department Of Internal Medicine "F" And Nephrology Division, Assaf
Harofeh Medical Center, Zerifin, Beer Yaakov, Israel. Internal6@Asaf.Health.Gov.Il
Hypomagnesemia and especially intracellular Mg depletion are frequent in
congestive heart failure (CHF). In many such patients furosemide treatment and
comorbidity with diabetes mellitus (DM) and/or renal dysfunction (RD) are common.
DM and furosemide may aggravate hypomagnesemia, while renal dysfunction may
cause hypermagnesemia. Since normal serum Mg concentration may coexist with
intracellular Mg aberrations, we studied peripheral blood mononuclear cell Mg
content in four relevant normomagnesemic patient subgroups. Patients: Included
were 71 (33 males, 39 females, age 72.3±9.7 years) furosemide treated patients with
symptomatic CHF, classified as follows: A. 12 patients without DM or RD; B. 21 with
DM; C. 14 with RD; D. 24 with DM and RD.
RESULTS: Whole group Intracellular Mg content was 1.66±0.29 mg/mg cell protein.
Individual values in the 4 subgroups A; B; C and D were 1.69±0.37; 1.81±0.26;
1.59±0.29; 1.57±0.22 respectively. When compared with DM subgroup, respective Pvalues
for subgroups RD, RD+DM and no DM/RD were 0.04, 0.002 and 0.6 (N.S).
Thus, only RD is associated with a significantly lower Mg content, and overrides the
tendency, albeit not reaching statistical significance, of DM for elevated values.
CONCLUSIONS: Within a group of normomagnesemic furosemide treated CHF
patients, intracellular Mg content differed in various subgroups in parallel with
comorbidities. RD as opposed to DM is associated with lower Mg content.
Assessment of intracellular Mg in relevant patient subgroups is essential for
evaluation of risks and preventive measures.

28
Long-Term Outcome of Intravenous Magnesium Therapy in Thrombolysisineligible
Acute Myocardial Infarction Patients
Michael Shechter, MD, MA; Hanoch Hod, MD; Babeth Rabinowitz, MD; Valentina
Boyko, MS; Pierre Chouraqui, MD From the Heart Institute, Sheba Medical Center,
Tel-Hashomer, and Sackler School of Medicine, Tel-Aviv University, Israel.
shechtes@netvision.net.il
Background. The use of magnesium in treating acute myocardial infarction (AMI)
remains controversial. We previously reported that, compared to placebo, 48-hour
intravenous magnesium sulfate (22 g) given to 194 consecutive thrombolysisineligible
AMI patients, significantly reduced in-hospital mortality rate with an odds
ratio of 0.21 (0.07-0.64) (p=0.01).
Methods and Results. The aim of our current study was to analyze long-term
survival and cardiac function of the same study population. Rest radionuclide
ventriculography tests for left ventricular ejection fraction (LVEF) were assessed in
surviving patients up to completion of follow-up. All-cause mortality was significantly
lower after a mean 4.8-year follow-up (18 vs 33 patients, p

29
In vitro application of endotoxin to thoracic aortas from magnesium-deficient
rats enhances vascular hyporeactivity to phenylephrine
A. Miyamoto, K. Moriki, S. Ishiguro and A. Nishio, Department of Veterinary
Pharmacology, Faculty of Agriculture, Kagoshima University, Kagoshima, Japan.
miyamoto@vet.agri.kagoshima-u.ac.jp
Endotoxin-induced vascular hyporeactivity to phenylephrine (PE) is well described in
rat aortas, but has not been investigated in those from magnesium (Mg)-deficient rats
in vitro. Segments of thoracic aorta from control and Mg-deficient rats were incubated
in culture medium for 6 h in the presence or absence of bacterial lipopolysaccharide
(LPS; 0.001-10ug/ml). Contractions to PE were measured with or without a nitric
oxide synthase (NOS) inhibitor (1400W;1,5,10uM), a guanylyl cyclase inhibitor
(ODQ;1,5,10uM), or a potassium channel inhibitor (TEA;1,10mM). LPS induced
hyporeactivity in a concentration-dependent manner under relatively low
concentrations (0.001-0.1ug/ml), however, there was no significant difference among
0.1, 1 and 10ug/ml. LPS-induced hyporeactivity was not significantly affected by
endothelium-denudation. The hyporeactivity was enhanced in thoracic aortas from
Mg-deficient rats by LPS (0.01, 0.1 and 1ug/ml). LPS (1ug/ml)-induced hyporeactivity
was reversed with 1400W, ODQ or TEA in both aortas in a concentration-dependent
manner, however the degree of reversal was weaker in Mg-deficient rat aorta than in
control rat one. Inducible NOS (iNOS) mRNA level was increased by LPS (0.1ug/ml),
and the increment was significantly high in Mg-deficient rat thoracic aorta. From
these results it is clearly demonstrated that LPS-induced vascular hyporeactivity to
PE is enhanced in thoracic aorta from Mg-deficient rats, and suggested that LPSinduced
NO production might contribute to the enhancement via stimulation of NOcyclic
GMP-potassium channel pathway.

30
Case for subcutaneous magnesium product for space missions
William J. Rowe M.D. Former Assist. Clin. Prof. Med. Medical College of Ohio at
Toledo, USA. rowerun@aol.com
Space flight (SF) cardiovascular (CV) complications are caused by microgravity,
hypokinesia, and radiation, particularly beyond earth orbit, with all 3 conducive to
oxidative stress. Pharmaceuticals, except for emergencies, appear in general
contraindicated, because of unpredictable side effects related to malabsorption (M)
and potential hepatic and renal impairment. Magnesium (Mg) depletion and
elevations of cytokines (interleukin 6) occur on SF, conducive to self-sustaining
vascular inflammation mechanisms. There are potential endothelial injuries (EI) and
reductions of cyclic GMP (a second messenger of nitric oxide) (NO) and elevated
urinary excretion of C-peptide (insulin resistance)(IR). Recently vascular endothelial
growth factor (VEGF) reductions have been shown, which may be a result of SF -
related thrombocytopenia since platelets (P) are the major source of VEGF and P
may be derived partially from NO. Both VEGF and Mg are vital for angiogenesis,
endothelial function and reendothelialization. Insulin is necessary for VEGF
expression. To prevent SF-related CV complications in the presence of IR and M and
with the potential for renal insufficiency, closely monitored subcutaneous (SC) Mg
must be administered. The dosage can be monitored by sublingual intracellular Mg
assays. A SC Mg reservoir device should be developed, which can be replenished
before extra- vehicular activities (EVA) and which must be reliable despite vigorous
movements during EVA, lasting up to 8 hours. This could also be protective against
decompression sickness and EVA-related 100% oxygen requirements prior and
during this activity, both of which predispose to EI. CONCLUSIONS To prevent CV
complications with EI and because of SF-related pharmaceutical constraints and M, it
is vitally important to develop a SC Mg product and a reliable device for its delivery,
with close monitoring of the dosage.

31
Relation of hypertension and atherosclerosis: 1H- and 31P-NMR spectroscopy
of free Mg, plasma membrane and circulating lipids in hypertension
Lawrence M. Resnick 1 , Raj K. Gupta 2 . 1 New York Presbyterian Hospital / Cornell U.
NY, NY, USA and 2 Albert Einstein College of Medicine, Bronx, N.Y.
LMResnick@aol.com
To study the linkage between lipids and ions in hypertension we utilized 1H- and
31P-NMR spectroscopy to measure RBC intracellular free Mg (Mgi), compared with
the degree of fatty acid unsaturation (DB/CH3), delta-5-unstaurated fatty acids (delta-
5/CH3, including arachidonate, DHA, EPA), fatty acid chain length (CL), and relative
cholesterol content (Chol/PC) in lipids extracted from RBC membranes and plasma
of normal (Nl), untreated (EH) and treated (EH-Rx) essential hypertensive subjects.
Compared to Nl, EH had lower Mgi (168 vs 202 mcM, p=0.01), and greater chol/PC
content (p=0.03), with lower fatty acid CL (p=0.05), delta-5/CH3 (p=0.02) and
DB/CH3 in membranes (p=0.006) and plasma (p=0.008). DB/CH3 (r=0.795, p=0.001)
and chol/PC (r=0.566, p=0.04) in plasma vs membrane fractions were closely
related. Altogether, Mgi was directly related to CL (r=0.711, p=0.002) and DB/CH3
(r=0.54, p=0.02), and inversely related to chol/PC (r=-0.563, p=0.01). Clinically, DBP
was inversely related to Mgi (r=-0.622, p=0.002) and DB/CH3 (plasma-r=0.735,
p=0.005; membranes r=-0.8, p=0.001); and positively related to chol/PC (plasmar=0.689,
p=0.02). These data demonstrate significant linkage of deficient Mgi with
shorter, more saturated fatty acids and increased cholesterol content in EH. These
data suggest how altered lipid metabolism may influence Mg, an established
determinant of arterial stiffness and blood pressure, and thus also the relation of
atherosclerosis and hypertension.

32
Functional effects of magnesium and statin pharmaceuticals compared
Andrea Rosanoff and Mildred S. Seelig, M.D., Independent Scholar, Pahoa, Hawaii,
USA. nutrient@gte.net
Mg2+-ATP controls the rate-limiting enzyme in the cholesterol biosynthesis sequence
targeted by the statin pharmaceuticals. In cholesterol biosynthesis, a series of
enzymatic reactions convert HMG-CoA to cholesterol. The rate-limiting reaction of
this pathway is the enzymatic conversion of HMG CoA to mevalonate via HMG CoA
Reductase (Reductase). Statin drugs inhibit Reductase and lower LDL cholesterol
levels by 35 to 65%. Magnesium supplementation can lower total cholesterol by 6 -
23%, and LDL-cholesterol 10 - 18%. Statins also reduce total, cardiac and stroke
mortality as well as incidence of heart attacks, angina and other nonfatal cardiac
events. These pleiotropic effects derive from statins' reduction of mevalonate
production. Reduced mevalonate production by statins inhibits proliferation,
inflammation and migration of vascular smooth muscle cells and macrophages by
decreasing interleukin-6 and inducible nitric oxide synthase, effects that are
completely blocked by exogenous mevalonate. As a result, mevalonate reduction by
statin drugs promote plaque stabilization and regression, reduce inflammation and
coagulation, and improve endothelial and anti-oxidant function in smooth muscle
cells. Statins reduce incidence of coronary events by 24-34%, stroke by 10-31%.
Mg2+-ATP deactivates Reductase. With a cellular magnesium deficiency, active
Reductase is less apt to be deactivated, and more cholesterol and more mevalonate
synthesized. Magnesium is a co-factor for other lipid enzymes, such as LCAT.
Magnesium supplements raise HDL-cholesterol by 4 - 11% and lower triglycerides by
10 - 31%, actions not as reliably effected by statin, which only act on Reductase.
Magnesium supplements achieve all beneficial corrections of dyslipidemia without
adverse side-effects. Statin drugs mainly lower LDL- cholesterol, and in 2%
adversely affect muscle and/or liver. Statins inhibit the enzyme responsible for
mevalonate and cholesterol production while normal cellular magnesium levels
regulate this pathway important for biosynthesis of CoQ10, bile salts, steroidal
hormones and other substances. Magnesium at optimal cellular concentration is well
accepted as a natural calcium channel blocker. More recent work shows that
magnesium also acts as a statin.

33
Role of magnesium in pathogenesis of essential hypertension
G. Sur and Oana Maftei. 2 nd Pediatric Clinic Cluj-Napoca, Romania
University of Medicine and Pharmacy “Iuliu Hatieganu”, Cluj-Napoca, Romania.
oana_maftei@yahoo.com
Magnesium seems to be involved in the pathogenesis of essential hypertension by
decreasing the sympathetic reactivity, cardiac excitability and vascular tone,
contractility and reactivity. The authors of this study proposed to assess the
reduction of blood pressure values after magnesium supplementation in children with
essential hypertension. We made a retrospective study on 17 children of 10 years to
18 years diagnosed with essential hypertension in 2 nd Pediatric Clinic of Cluj-Napoca,
across 2 years (January, 2001 – December, 2002). In these children we excluded all
the causes of a secondary hypertension. There were 8 children (47%) with a deficit
of magnesium. We administered magnesium (Magnerot 15 mmol/24 hours), to all the
children with essential hypertension, for 2 months, in association with a saltrestrictive
diet and physical exercises. To those patients without hypomagnesiemia,
we gave just half of the dose of magnesium. Most patients with primary hypertension
were adolescents (there were 9 patients over 15 years). Their blood pressure values
were just 5-10 mmHg over the normal values for their age, but because their familial
history was positive for hypertension, we recommended magnesium, diet and sport.
There were 7 girls (41%) and 10 boys (59%) with hypertension. Eight of these
children had hypomagnesiemia (3 girls and 5 boys). After 10 days of magnesium
supplementation, diet and sport, both systolic and diastolic blood pressure values
decreased by 10mmHg, palpitations disappeared, sleep became still. The
improvement of the patients’ status and the decline of blood pressure values were
more evident in those patients with hypomagnesiemia. The role of magnesium
supplementation in patients with high blood pressure is beneficial in most cases. But
it is also important to follow a salt-restrictive diet and to practice physical exercise.

34
Cardiovascular and skeletal benefits of estrogen replacement therapy in
healthy postmenopausal women; risks intensified by ignoring importance of
magnesium
Mildred S. Seelig, Burton M. Altura, and Bella T. Altura. Department of Physiology
and Pharmacology, State University of New York, Downstate Medical Center,
Brooklyn 11203, USA. mgseelig@comcast.net
Numerous controlled clinical investigations have found that estrogen replacement
therapy (ERT) alone or combined with progestogen: hormone replacement therapy
(HRT), protects postmenopausal women against cardiovascular disease (CVD) and
osteoporosis, to which they are much more prone than are young women. The
Women’s Health Initiative (WHI) of the NIH, planned to run for six years (to obtain
statistically significant benefits to risks data) was ended early because there were
more coronary events, strokes and other embolic phenomena, as well as invasive
breast cancers in HRT women than in controls (about eight more each). Women
receiving HRT had several fewer hip fractures than did those taking placebo.
Paradoxical effects of female sex hormones on CVD are partially ascribable to the
cardioprotective effects of estrogen, yet adverse microvascular actions, but more to
their effects on magnesium distribution - lowering its levels in blood, but increasing
that in soft tissues. Magnesium exerts beneficial effects by increasing resistance to
arrhythmias and coronary arterial constriction, and exerting antioxidant activities -
partly by reducing formation of reactive oxygen species. At optimal levels magnesium
inhibits the coagulation cascade and platelet aggregation/ adhesion. Decreased
plasma magnesium contributes to coronary vasospasm and intensifies estrogen’s
induction of thrombotic events, which has complicated former use of high dose
estrogen in men with prostatic cancer, and to achieve womens’ cardioprotection in
men with coronary disease. Magnesium’s protection against oral estrogencontraceptive
induction of thromboses was first noted in 1970. Estrogen and
progesterone have recently been shown to affect magnesium and calcium differently,
depending on hormonal levels. Estrogen’s shift of magnesium from blood to bone
matrix is contributory to its anti-osteoporotic effect. Osteoporosis is a risk in patients
with alcoholism or diabetes, which are associated with severe Mg depletion.

35
Osteoblastic cell growth as a function of Ca 2+ /Mg 2+ ratio
O. Theodorakopoulou 1 , D. Deligianni 2 and J. Anastassopoulou 1, 1 National Technical
University of Athens, Chemical Engineering School, Radiation Chemistry and
Biospectroscopy, Zografou Campus, 15780 Zografou, 2 University of Patras,
Department of Mechanical & Aeronautical Engineering, 26500 Patras, Greece.
theophan@central.ntua.gr
Magnesium ions play a crucial role in living cells and are involved in almost all living
processes. Magnesium ions exist in aqueous solutions and in biological systems as
hydrated hexaaquated cations Mg(H2O)6 2+ . Magnesium ions stabilize the structure of
DNA in a conformation, which depends on Mg 2+ concentration 1 . Increasing the
concentration of magnesium ions in DNA inverts the B- form to Z-DNA 2,3 . Both cell
growth and carcinogenesis are magnesium concentration dependent. Magnesium
deficiency antagonizes tumor implication and inhibits growth of tumors in rats 4 . This
could be due to a disturbance in magnesium homeostasis in carcinogenesis. It was
found that hypomagnesemia is associated with disturbance in Ca 2+ -homeostasis 4 .
Thus, in this work we used human osteoblastic bone marrow cells to examine the
influence of the magnesium and calcium ratios to the bone growth. The process of
the cells was compared with unmodified control cell. The cells were monitored for
differences in their proliferation, morphology and viability. It was found that the
survival time was highest for a ratio [Ca 2+ /Mg 2+ ]=3:1. The results on the mortality of
the cells as a function of the ratio Ca 2+ /Mg 2+ will be presented and discussed in the
meeting.
References
1. J. Anastassopoulou and T. Theophanides, Magnesium - DNA interactions
and the possible relation of magnesium to carcinogenesis. Irradiation and free
radicals Oncology and Heamatology, 42, 79-91, 2002
2. T. Theophanides and H.A. Tajmir-Riahi, “Flexibility of DNA and RNA upon
binding to different Metal Cations. An Investigation of the B to A to Z
Conformational Transition by Fourier Transform Infrared Spectroscopy” J.
Biomolecular Structure and Dynamics Vo 2, 995-1004 (1985).
3. J. Anastassopoulou and T. Theophanides, Free radicals and magnesium
concentration effects on DNA structure, in Advances in Magnesium:
Pharmacology, Metabolism and Nutrition,Ed. J.F. Escanero, John Lybbey,
Chapt 5, 1-5, In press.
4. KS Kasprzak, Effects of calcium, magnesium zinc and iron on nickel
carcinogenesis: Inhibition versus enhancement, in Cytotoxic, Mutagenic and
carcinogenic potential of heavy metals related to human environment,
Dordrecht: Kluwer Academic Publishers, 93-106 (1997).

36
Bone mineral density, serum albumin and serum magnesium
Noboru Saito, Naoto Tabata, Toshiaki Setoguchi, Saburou Saito, Harumi Sayama 1),
Toshiko Kannagi 1) Dept. of Internal Medicine and Lifestyle-related diseases,
Miyazaki Medical Center Hospital, Miyazaki, Japan, 1) Cardiovascular Division,
Kyoto Prevention Medical Center, Kyoto, Japan. hit.yama@d6.dion.ne.jp
This study aimed to investigate how bone mineral density (BMD) was related to
serum albumin or magnesium (Mg).
1) In this study 217 subjects (159 men, 58 women) aged mean 57.7 years were
recruited, who were ambulant and diagnosed about diabetic type through 75gOGTT.
BMD was measured by DXA method using lumbar vertebra and expressed as
percentages against young adult mean (YAM).
2) 122 inpatients (36 men, 86 women) aged mean 80.1 years were adopted, who
suffered from cerebrovascular diseases, hypertension and so on. BMD of forearm
bone was measured by DXA method.
In these above cases fasting blood sample was obtained early in the morning for
biochemical data. Serum Mg was measured with colorimetry of xylidil blue method.
1) When the percentages of BMD against YAM became low, serum albumin and Mg
decreased significantly in male subjects, and body mass index (BMI) decreased in
both sexes. During observation period of mean 22.7 months BMD and serum
albumin increased significantly.
2) In almost elderly inpatients, BMI, serum albumin and Mg decreased significantly,
when the percentage of BMD became low.
From these findings it was concluded that BMD could be related to serum albumin
and Mg.

37
Magnesium, magnesium deficiency and interaction with aminoglycoside and
quinolone antibiotics
Prof. Dr. Juergen Vormann, Institute for Prevention and Nutrition, Ismaning/Munich,
Germany. vormann@ipev.de
The interaction with magnesium respectively magnesium deficiency is of
considerable interest in the use of various antibiotics. The most important side effects
of aminoglycoside antibiotics are oto- and nephrotoxicity, occurring in over 2% of
patients. In animal experiments it could be shown that magnesium deficiency
increased the ototoxic effects of gentamicin significantly. Aminoglycoside antibiotics
themselves induce magnesium deficiency by increasing renal magnesium wastage.
Quinolone antibiotics have the potential to cause joint cartilage damage in juvenile
animals. Due to these arthropathogenic effects use of these antibiotics is
contraindicated in children. Lesions resembling those observed after quinolone
treatment can also be induced by feeding immature rats a magnesium deficient diet
alone. Chondrotoxicity and tendopathy is additionally increased by combining
magnesium deficiency and quinolone treatment. On the other hand supplementation
with magnesium diminishes quinolone-induced arthropaty in growing rats. In murine
limb bud cultures quinolones impaired limb development; an effect that was
enhanced by culture in magnesium-deficient medium. The toxicity of quinolones is
probably caused by forming stable chelate complexes with magnesium ions, leading
to a relative magnesium deficit in the weakly perfused connective tissues of the
musculoskeletal system.

38
Magnesium in sports
Frank C. Mooren, MD, Institute of Sports Medicine, Muenster University-Hospital,
Muenster, Germany. mooren@uni-muenster.de
Although the regulation of magnesium (Mg2+) during exercise has attracted
scientist's interest for many years our view is still far from being complete. Two
reasons are largely responsible for this situation. First, most studies have focused on
the magnesium concentration in the extracellular compartment only. Secondly, most
of the currently available data on the regulation of Mg2+ during exercise are
unfortunately based on determination of total magnesium and did not include
measurements of ionized Mg2+ levels. Nevertheless data available so far clearly
demonstrate that exercise results in alterations of Mg2+ homeostasis, which seems
to depend on type, duration and intensity of exercise. After short-term, high-intensity
exercise the majority of studies indicated an increase of extracellular Mg2+, which
seemed to be caused predominantly by an exercise induced decrease in plasma
volume. The exercise-induced changes in ionized extracellular magnesium were
measured so far only in one study. After anaerobic exercise an increase in ionized
Mg2+ was found, which we could confirm in our studies. After prolonged submaximal
exercise most studies reported a hypomagnesaemia. It seems unlikely that sweat
Mg2+ losses and/or enhanced renal Mg2+ excretion account for this decrease in
plasma Mg2+. Indeed, during moderate exercise a slightly decreased urine excretion
of Mg2+ was reported. Some authors suggested that during prolonged exercise a
shift of Mg2+ into the cellular compartment occurs. In contrast, longitudinal and
cross-sectional studies suggest that intensive training may be followed by Mg2+
depletion and that athletes are prone to Mg2+ deficiency.

39
Cytosolic free [Mg2+] in the human calf muscle in different metabolic
conditions
Associate Professor Stefano Iotti, University of Bologna, Bologna, Italy.
iotti@med.unibo.it
Accurate knowledge of intracellular [Mg2+] is crucial for a deeper understanding of
cellular bioenergetics, and detecting changes of [Mg2+] that may alter critical
regulatory mechanisms causing abnormal metabolism. In the skeletal muscle,
variations of pH, phosphocreatine and inorganic phosphate concentrations influence
the complex multi-equilibrium system of the molecular species binding magnesium.
As a consequence [Mg2+] can change considerably in different metabolic conditions
such as rest, exercise and recovery. We compared the [Mg2+] assessed in vivo in
the human calf muscle by 31P MRS and calculated by a computer simulation in
different metabolic conditions. We studied 42 controls by a G.E. 1.5 T Signa System.
[Mg2+] was assessed from the chemical shift of beta-ATP (1). Computer simulation
was performed, by HYSS (HYperquad Simulation and Speciation) (2). We found by
31P-MRS a mean resting [Mg2+] of 0.32 mM. This value was used to calculate by
HYSS the total amount of magnesium in the muscle cells, which was 7 mM. The in
vivo assessment showed that during exercise and recovery [Mg2+] remarkably
changed, due to the predominant effect of pH. The plot of [Mg2+] as a function of pH
showed an exponential pattern with a sharp increase of [Mg2+] below pH 6.5.
Simulation by HYSS showed a much smaller increase of free [Mg2+] in the same pH
region. Results show that our model well describes the in vivo [Mg2+] of muscle cells
under different metabolic conditions, suggesting the existence of more Mg-binding
sites releasing Mg2+ at low pH.
1 Iotti S. et al. MRI 18, 607, 2000
2 Gans P. et al. Talanta 43, 1739, 1996

40
A functional biological marker is needed for diagnosing magnesium deficiency
Kay B. Franz, Department of Nutrition, Dietetics and Food Science, Brigham Young
University, Provo, Utah 84602, USA. kay_franz@byu.edu
Functional biological markers (FBM) are now available for many nutrients and are
used as nutritional status markers. Examples include ferritin for iron stores, serum
transferrin receptor for early iron deficiency, methylmalonic acid for vitamin B-12
deficiency, and selenium-dependent glutathione peroxidase for selenium deficiency.
At this time we do not have FBM for Mg beyond the measurement of total or ionized
Mg in serum, plasma, cellular components, urine or Mg retention from a load test.
FBM for Mg would reflect changes in biochemical processes where Mg is involved.
Sources of FBM are products or precursors of enzymatic processes that can be
measured in serum and plasma or the activity of a Mg-requiring enzyme in a blood
cell. FBM for Mg need to be identified and evaluated in both animals and humans,
with a determination of possible factors that may affect the reaction and the FBM
concentration. Some fundamental processes and possible FBM for Mg include the
following: pumps in the plasma membrane, Na/K ATPase; platelet activity and
thrombus, thromboxane B2; acute phase protein, C-reactive protein; and endothelial
damage, endothelin-1. Other possible FBM need to be identified. Activity of
erythrocyte Na/K ATPase is decreased in Mg deficiency, while the serum/plasma
concentrations of thromboxane B2, C-reactive protein, and endothelin-1 have been
manipulated by Mg deficiency or Mg supplementation. Measurement of Na/K
ATPase is laborious, while kits are available for each of the other three. These
possible FBM need to be carefully considered.

41
The relation of birth weight to intracellular magnesium of cord-blood platelets
Junji Takaya, Fumiko Kotera and Yohnosuke Kobayashi, Department of Pediatrics,
Kansai Medical University, Moriguchi, Osaka, Japan. takaya@takii.kmu.ac.jp
Magnesium (Mg2+), the second most abundant intracellular cation, is a critical
cofactor in numerous enzymatic reactions. Diabetic patients and obese subjects are
reported to have an intracellular Mg2+ ([Mg2+]i) deficiency. Many epidemiological
studies have disclosed that restricted fetal growth has been associated with an
increased risk of insulin resistance in adult life. Our purpose was to determine the
correlation between birth weight and [Mg2+]i. The study group consisted of 58 infants
with gestational ages ranging from 36-41 weeks, and birth weights ranging from
1,798-4,030 g. The subjects were 13 infants with intrauterine growth retardation
(IUGR) and 45 infants with normal birth weight. By using a fluorescent probe, magfura-2,
we examined the basal and insulin-stimulated [Mg2+]i of platelets in the cord
blood. The mean basal [Mg2+]i, but not plasma magnesium, was lower in IUGR than
in the control group ( 335±162 umol/L vs. 487±184 umol/L, p

42
Balance of Mg positively correlates with that of Ca
Mamoru Nishimuta(1), Naoko Kodama(1,2), Eiko Morikunii(1), Yayoi H. Yoshioka(1),
Hidemaro Takeyama(1,3), Hideaki Yamada(1,4), Hideaki Kitajima(1,5) and
Kazumasa Suzuki(1), (1)The National Institute of Health and Nutrition (Tokyo) Japan,
(2)Medical University of Yamanashi, (3)Nagoya City University, (4)Mimasaka
Women's University, (5)Taisho Pharmaceutical Co. Ltd. nisimuta@nih.go.jp
From 1986 to 2000, 109 volunteers (23 males, 86 females), ranging from 18 to 28
years old, took part in mineral balance studies after written informed consent was
obtained. The duration of the study periods ranged from 5 to 15 days, with 2-4 days
of adaptation. Foodstuffs used in each study were selected from those commercially
available. The minerals present in diet, feces, urine and sweat were measured by
atomic absorption spectrophotometer (Ca, Mg) or spectrophotometer (P). The dietary
intakes of Ca, Mg and P ranged from 4.83-23.58, 2.44-7.83, and 13.46-45.69
mg/kgBW/d, respectively. Dietary intake (Intake) of Ca was positively correlated to
apparent absorption (A. A.) (r2=0.425), which was also correlated with urine
excretion (Urine) (r2=0.327) and balance (Bal) (r2=0.382). Intake of Ca was slightly
but significantly correlated with Bal (r2=0.036, p=0.048). Intake of Mg was positively
correlated to A. A. (r2=0.451), which was also correlated with Urine (r2=0.486) and
Bal (r2=0.349). However, intake of Mg was not correlated with Bal. Intake of P was
positively correlated with A. A. (r2=0.959), which was also correlated with both Urine
(r2=0.908) and Bal (r2=0.135). Intake of P was slightly but significantly correlated
with Bal (r2=0.103, p=0.0013). Values of Intakes of Ca, Mg and P when balance is
equal to zero were 11.752, 4.395 and 22.584 mg/kgBW/d, respectively. Intakes of
Mg and P correlated negatively with their respective A. A. rates (%)(r2=0.120 for Mg,
r2=0.109 for P). Balance of Ca was positively correlated with that of Mg, (r2=0.541).

43
Developed determination method of ultra-trace elements and ultra-trace
element levels in plasma of rat fed low magnesium diet
Mieko Kimura, *Kazuto Honda, Atsuko Takeda, Masayo Imanishi, Takahisa Takeda,
Takeda Research Institute of Life Science, Kyoto 600-8813 Japan, *Perkin-Elmer
Japan Co. Ltd., Suita Osaka 564-0051 Japan. mkimura@mbox.kyoto-inet.or.jp
The mineral concentrations in plasma are unchangeable, for the most part, by
homeostatic mechanisms and not always reflected in the nutritional status. For that
reason, nutritional assessment is difficult and another biological value is sometimes a
good indication. Recently, determination of ultra-trace element concentration has
become possible by improvements in the function of the measuring instrument. A
determination method for trace and ultra-trace element in the rat plasma without pretreatment
using an inductively coupled plasma mass spectrometry (ICP-MS Elan-
6000, Perkin Elmer,Co.,USA) was developed and basic data of trace element
concentration was studied for the first time. To investigate 10 trace and ultra-trace
elements status in low magnesium, 12 male Wistar young rats (3-weeks-old) were
divided into two groups and fed a normal diet or low magnesium diet for 4 weeks.
The plasma was diluted by ultra pure water (TAMAPURE-AA:Tama Chemical
Co.,Japan). Ten ultra-trace element (As: 75, Se:82, Rb:85, Sr:87, Mo:95, Ba:137,
Ta:181, Ir:193, Au:197, Pb:208) concentrations in the plasma were measured with
ICP-MS Elan 6000. As, Rb, Sr, Ta, Ir and Au concentrations were significantly high in
the plasma of low magnesium rats compared to those in the normal rats. Essentiality
for mammals was certified for Se, Rb, Mo, As and Pb, and not yet for Sr, Ba, Au, Ir
and Ta. This result proved that ultra-trace element concentrations in plasma was
changeable by nutritional conditions and possibilities to be finding new physiological
function and to be prove essentiality for mammal in these elements were suggested.

44
Magnesium and cancer in clinical practice (update).
Amid Reba*, MD, FACN, François Goldwasser**. MD, PhD. Hospital Cochin
Universite Paris V, 27 rue du faubourg St Jacques 75014 Paris France.
AMIDREBA@AOL.COM
The regulation of the cell cycle control and the apoptosis, of the DNA repair, depends
on many enzymatic reactions which are modulated by magnesium. As a
consequence, the relations between magnesium and cancer were the topic of
numerous studies. In vitro, its role on cell proliferation is negligible and depends on
the model investigated and on the experimental conditions. However, the Mg has a
protective effect on some DNA lesions, and can have carcinogenic inhibition in some
models. Otherwise, from animal studies, low magnesium diets do not inhibit growth
of tumor xenograft. Hypomagnesemia may become symptomatic in cancers.
Cytotoxic chemotherapy frequently induces hypomagnesemia and renal magnesium
loss through its renal tubular toxicity. The clinical studies show that it can be
dangerous if no corrective measures of this hypomagnesemia are initiated. Mg
supplementation could compensate or prevent the effects of anticancer
chemotherapy. In conclusion, the relationship between magnesium and neoplastic
disease is rather complex and not directly applicable in clinic. On the other hand, it is
demonstrated that the correction of hypomagnesemia is a therapeutic measure
susceptible to improve the treatment safety and the patients comfort.

45
Regulation of intracellular magnesium content during cellular proliferation
S. Fasanella, A. Torsello, B. Tedesco, A. Cittadini and F.I. Wolf. Institute of General
Pathology and Giovanni XXIII Cancer Research Center, Catholic University of
Sacred Heart School of Medicine, Rome Italy. fwolf@rm.unicatt.it
It is well known for decades that magnesium (Mg) availability is required for
proliferation, but the underlying regulatory mechanism has not been described in
detail. In order to investigate this important event, we measured intracellular Mg
content during cell growth in culture under different conditions. To this aim we used
immortalized mammary
epithelial cells (HC11) chronically grown in non-physiological high and low
Mg concentrations (45 mM high Mg and 0.025 mM low Mg). We observed that
during the exponential growth of the cells in culture, Mg content increased
by about 6 fold in control and high Mg media. In low Mg medium, the increase of
intracellular Mg, even though smaller, was also statistically significant. The increase
of intracellular Mg did not depend on adenine nucleotide pools but rather paralleled
DNA duplication. Intracellular Mg content of cells grown in non-physiological Mg
concentrations were balanced by membrane potential and pH: hyperpolarization
associated with unfavourable transmembrane chemical Mg gradient and reduced pH
associated with high transmembrane chemical Mg gradient. These data suggest that
intracellular Mg content is modulated by Mg-requiring metabolic activities, such as
DNA duplication, and also by the balance of different ionic species. We also
investigated how Mg moves through plasma membranes in the presence of modified
electrochemical
gradients by measuring uptake of the stable isotope 25Mg. At present, our
data do not rule out the role of specific transport mechanisms.

46
Magnesium, insulin resistance and body composition in healthly
posmenopausal women
Laires, M.J. 1 ; Moreira, H. 2 ; Monteiro, C.P. 1 ; Sardinha, L. 3 ; Veiga, L. 1 ; Gonçalves, A. 3 ;
Bicho, M. 4 1- Laboratório de Bioquímica, Faculdade de Motricidade Humana, U.T.L. 2-
Universidade de Trás-os-Montes e Alto Douro 3- Laboratório de Exercício e Saúde,
Faculdade de Motricidade Humana, U.T.L. 4- Laboratório de Genética, Faculdade de
Medicina de Lisboa, U.L. mjlaires@fmh.utl.pt
This study aims to associate body composition, magnesium levels and insulin
resistance. In 76 Caucasian sedentary postmenopausal healthy women (50-77
years) were evaluated: red blood cell's (RBC-Mg) and plasma (P-Mg) magnesium,
glucose, insulin, body mass index (BMI), body fat percent (BF%), abdominal fat (AF)
and fat free mass (FFM). Glucose intolerance was derived from fasting glucose and
insulin by the formula for the HOmeostasis Model Assessment (HOMA). Women
were divided according to BMI>25 (obese-GO) and BMI

47
Clinical efficacy of magnesium supplementation in patients with type 2
diabetes
K. Yokota, *M. Kato, **H. Li, M. Shiraishi, T. Hayakawa, **T. Kikuta, ***F. Lister,
****H. Akiyama, S. Kageyama and N. Tajima, Jikei University School Of Medicine,
Minato-Ku, Tokyo, Japan, *Kato Medical Clinic, **Matier Co.,Ltd., ****Srl Co.,Ltd.
Tokyo, Japan. ***WA Salt Supply, Australia. Yokota@jikei.ac.jp
Aims, Subjects and Methods. The effects of magnesium (Mg) supplementation on
type 2 diabetes were examined. Nine type 2 diabetic patients (mean age: 51.6 yrs)
with stable glycaemic control were enrolled. Water from a salt lake with a high Mg
content (7.1%) (MAG21) was used for supplementation after dilution with distilled
water to 100 mg/100 ml, and 300 ml/day was taken for 30 days.
Results. The serum Mg concentration, urinary Mg concentration, and urinary Mg
excretion rate increased significantly after supplementation. Fasting immunoreactive
insulin decreased significantly, as did HOMA-R (both p

48
Post-cholecystectomy syndrome and magnesium deficiency
P. J. Porr, J. Szántay, M. Rusu, 3rd Medical Clinic, Cluj-Napoca, Romania.
pjporr@umfcluj.ro
In 20-30% of cholecystectomized patients a biliary syndrome reappears after some
weeks or months, called post-cholecystectomy syndrome (PCES). The etiology of
this is in certain cases an anatomic one (choledochal lithiasis or stricture, obstructive
papillitis, pancreatic duct stenosis a.o.), but there are a lot of cases in which all
organic causes are excluded. The aim of this study was to analyze the correlation
between these functional disturbances and a magnesium deficiency (MD). We
analyzed 52 patients with PCES and MD, in which organic lesions of the remaining
bile ducts were excluded by imaging and endoscopic methods. MD was confirmed by
serum and erythrocytic low Mg levels. 82,6% of patients were women. The
substitution therapy was performed with Tiomag (Mg gluconate and methionine), B6
vitamin and Ca lactate for 6 weeks or more. In 50 patients the symptomatology of
PCES disappeared after this treatment. In 14 cases some symptoms appeared again
after a few weeks - months, but after repetition of the same therapy they disappeared
definitively. Our results demonstrate the dependence of PCES functional
manifestations on MD, especially the recurrence of symptoms, subsided again after
the Mg substitution therapy was resumed.

49
Effect of magnesium diets in ischemic stroke rats
Céline Demougeotº, Sylvie Bobillier Chaumont*, Claude Mossiatº, Christine Marieº
and Alain Berthelot*. º Laboratoire de Pharmacodynamie et Physiologie
Pharmaceutique, UFR SMP Dijon; * Laboratoire de Physiologie Pharmacologie
Nutrition Préventive Expérimentale, UFR SMP Besançon. alain.berthelot@univfcomte.fr
In order to study the impact of magnesium (Mg) dietary intake on the vulnerability of
the brain to ischemia, the photothrombotic ischemic stroke model was induced in rats
fed with low (0.015 %), normal (0.08 %) or high (0.32 %) Mg diet for 5-6 weeks.
Infarct volume and plasma total Mg level were measured 4h and 24h after irradiation.
As compared to normal dietary Mg intake, low Mg diet was associated with
decreased plasma Mg level at 4h and 24h and increased infarct volume at 24h-postirradiation.
Conversely, whereas high Mg diet increased plasma Mg level at 4h and
24h without accompanying hyperglycemia and hypotension, it failed to reduce infarct
volume. Variations in Mg dietary intake Mg level were not associated with
modification of plasma antioxidant capacity. However, plasma Mg level correlated
with plasma antioxidant capacity, at least in the early phase of ischemia (4h postirradiation).
Our data demonstrate that brains from rats fed with low Mg diet are more
susceptible to permanent focal ischemia. They may have important implications for
future clinical stroke trials.

50
A substance P antagonist increases brain intracellular free magnesium
concentration after diffuse traumatic brain injury in rats
Robert Vink, Maria I. Cruz*, James J. Donkin, Alan J. Nimmo† and Ibolja Cernak*
Department of Pathology, University of Adelaide, Adelaide SA, Australia;
*Department of Neuroscience, Georgetown University, Washington DC, USA;
†School of Pharmacy and Molecular Sciences, James Cook University, Townsville
QLD, Australia. ifc@georgetown.edu
Magnesium deficiency has been shown to increase substance P release and induce
a pro-inflammatory response that can be attenuated with the administration of a
substance P antagonist 1 . Neurogenic inflammation has also been implicated in
traumatic brain injury (TBI), a condition where brain intracellular free magnesium
(Mgf) decline is known to occur and has been correlated with functional outcome 2 .
We therefore examined whether a substance P antagonist restores brain intracellular
free magnesium concentration following TBI. Male, adult Sprague-Dawley rats were
injured using the Cernak rodent model of diffuse TBI 3 . At 30 min after injury, animals
were administered either 0.25 mg/kg i.v. n-acetyl tryptophan or equal volume saline.
Prior to and 4 h after induction of injury, phosphorus magnetic resonance spectra
were acquired using a 7-tesla magnet interfaced with a Bruker spectroscopy console.
Mgf was calculated from the chemical shift of the beta ATP. Prior to injury, Mgf was
0.51 ± 0.05 mM (SEM). By 4 hr after injury, Mgf had significantly declined to 0.27 ±
0.02 mM in saline treated animals. In contrast, animals treated with n-acety
tryptophan had a Mgf of 0.47 ± 0.06 mM at 4 h after injury, which was not significantly
different from preinjury values. There were no significant differences in pH between
the treatment groups. We conclude that any beneficial effect of a substance P
antagonist on functional outcome following TBI may be related to the improvement in
brain magnesium homeostasis induced by the compound.
1. Kramer, J.H., Phillips, T.M. and Weglicki, W.B. (1997). J. Mol. Cell. Cardiol. 29:
97-110.
2. Vink, R. and Cernak, I. (2000). Front. Biosci. 5: 656-665.
3. Cernak, I., Vink, R., Cruz, M.I. and Faden, A.I. (2002). J. Neurotrauma 19: 1397.

51
Amiloride increases neuronal damage after traumatic brain injury in rats
Renee J. Turner, Corinna van den Heuvel and Robert Vink. Department of
Pathology, University of Adelaide, Adelaide, SA, Australia.
Robert.Vink@adelaide.edu.au
It is well known that traumatic brain injury (TBI) decreases brain free magnesium
concentration, while administration of magnesium salts after TBI restores the
concentration and improves functional outcome 1 . In the presence of haemorrhage,
administration of exogenous magnesium salts exacerbates the injury process and
worsens outcome 2 . An alternative to administration of magnesium after injury may
be to prevent cellular loss of magnesium with the use of amiloride, which inhibits
Na/Mg exchange. In the present study, male, adult Sprague-Dawley rats were
injured using the impact acceleration model of diffuse TBI 2 and administered either
100µmoles/kg i.v. amiloride or equal volume 50% DMSO/saline at 30 min after injury.
Amiloride did not improve functional outcome (motor or cognitive outcome) after TBI
relative to vehicle treated controls. Histologically, treatment with amiloride
significantly increased hippocampal caspase-3 expression (apoptosis), axonal
swellings in the medulla, and the degree of neuronal dark cell change (cell stress) in
the cortex. Phosphorus NMR demonstrated that amiloride did not increase free
magnesium concentration after TBI. We conclude that amiloride is ineffective in
preventing magnesium loss after TBI when administered at 30 min after trauma.
Moreover, by administering amiloride after the TBI related magnesium decline has
already been initiated, it may exacerbate injury by, in part, inhibiting Na/Mg antiport
and preventing entry of magnesium back into the cell, but also by inhibiting other Na
linked transporters.
1. Vink, R. and Cernak, I. (2000). Front. Biosci. 5: 656-665.
2. Heath, D.L. and Vink, R. (2001) J. Neurotrauma 18: 465-469.

52
Propofol attenuates the neuroprotective effects of magnesium in experimental
traumatic brain injury
Tulin Erdem*, Damla Aktan*, Mehmet Kaya**, S.Murat Imer***, Nahit Cakar*, Lutfi
Telci*, Figen Esen*. *University of Istanbul, Istanbul Medical Faculty Department of
Anesthesiology & Reanimation, Istanbul-Turkey; **University of Istanbul, Istanbul
Medical Faculty Department of Physiology, Istanbul-Turkey *** University of Istanbul,
Istanbul Medical Faculty Department of Neurosurgery, Istanbul-Turkey.
erdentn@yahoo.com
OBJECTIVES: Propofol is a popular nonbarbiturate anesthetic agent. Its
neuroprotective effects are controversial. The neuroprotective effects of magnesium
salts have been documented. We aimed to examine the neuroprotective effects of
propofol alone and with magnesium on brain edema and blood-brain barrier (BBB)
breakdown after experimental traumatic brain injury (TBI) in rats.
METHODS: Experimental closed head trauma was induced on Spraque-Dawley rats
by allowing 450-gm weight falling from a 2-m height onto a metallic disc fixed to the
intact skull. Rats were assigned into four groups to receive intraperitoneally 1 ml/kg
saline in the control group (C, n=10), 10 mg/kg propofol in the propofol group (P,
n=10), 750 µmol/kg magnesium sulphate (MgSO4) in the magnesium group (M,
n=10), 10 mg/kg propofol and 750 µmol/kg MgSO4 in the magnesium-propofol group
(PM, n= 10) 30 minutes after TBI. Brain water content (BWC) and specific gravity
(SG), as indicators of brain edema were measured 24 hours after TBI. BBB
breakdown was evaluated quantitavely 24 hours after TBI by fluorometric assay of
Evans blue dye (EBD) extravasations.
RESULTS: The increase in BWC, the reduction in SG and EBD content in the group
P was statistically significant when compared to the group C. In the group PM, BWC
was significantly higher and SG was significantly lower than group M. EBD content in
the brain tissue was also significantly increased in the group PM when compared to
group M.
CONCLUSIONS: These experimental data have shown that although propofol has
neuroprotective effects on TBI, it is not as effective as magnesium, and it attenuates
the neuroprotective effects of magnesium sulphate on secondary injury factors
following traumatic brain injury.

53
Simultaneous monitoring of extracellular magnesium and zinc in gerbil cortex
during focal cerebral ischemia by microdialysis-graphite furnace atomic
absorption spectroscopy
Professor FU-Chou Cheng, Department of Medical Research, Taichung Veterans
General Hospital, Taichung, Taiwan. vc1035@sinamail.com
The aim of this study was to develop a microdialysis-graphite furnace atomic
absorption spectroscopy (MD-GFAAS) method for monitoring dynamic changes of
extracellular magnesium and zinc in the cortex of gerbils subjected to focal cerebral
ischemia. Focal cerebral ischemia was produced in anesthetized gerbils by occlusion
of the right common carotid artery and the right middle cerebral artery for 1 h
followed by a 3-h reperfusion. Two microdialysis probes were inserted into both sides
of the cortex to simultaneously collect dialysates during cerebral ischemia and
reperfusion. Dynamic changes in these analytes, on ipsilateral and contralateral
sides of the brain, were assayed by MD-GFAAS. Optimal conditions and analytical
precision of GFAAS were studied in the present assay. The present study
demonstrated significant decreases in magnesium (50 % of baseline) and a transient
increases in zinc (195 % of baseline) on the ipsilateral side of cortex during cerebral
ischemia in the control group. Magnesium and zinc levels gradually returned to
baseline after reperfusion.

54
Determination of extracellular magnesium in brains of gerbils subjected to
cerebral ischemia by an on-line microdialysis and graphite furnace atomic
absorption spectrometry
Jen-Bin Lee. Department of Applied Chemistry, Providence University, Taichung,
Taiwan. pianofis@seed.net.tw
An in vivo microdialysis system was constructed for the determination of extracellular
magnesium levels in gerbil brain dialysates. The microdialysis hyphenated system,
consisting a microdialysis sampling device and a graphite furnace atomic absorption
spectrometer, was developed. The linearity of magnesium concentrations ranged
from 0.50 to 5.0 ppb with a detection limit of 0.03 ppb in the present assay. Each
assay was completed within 3 min. The intra- and inter-assay precision was within 5
%. A microdialysis probe was inserted into the right cortex of a gerbil subjected to a
focal cerebral ischemia. The focal cerebral ischemia was induced by an occlusion of
the right middle cerebral artery and the right common carotid artery for 60 min
followed by additional 180 min reperfusion. Dynamic changes of brain magnesium
levels were demonstrated during cerebral ischemic and reperfusion periods. During
cerebral ischemia, magnesium levels decreased significantly to about 50 % of
baseline in the cortex and returned to the baseline within 3 h of reperfusion.

55
Microdialysis coupled with graphite furnace atomic absorption spectrometry in
the determination of blood magnesium levels in gerbils subjected to cerebral
ischemia/reperfusion
Ming-Cheng Lin, Department of Medical Technology, Chung-Tai Institute of Health
Sciences and Technology, Taichung, Taiwan. vickyliu@sunrise.hkc.edu.tw
Microdialysis coupled with graphite furnace atomic absorption spectrometry system
was developed and employed for the dynamic monitoring of blood magnesium levels
in gerbils subjected to focal cerebral ischemia. A transient focal cerebral ischemia
was induced by the occlusion of the right middle cerebral artery and the common
carotid artery for 60 min followed by 60 min of reperfusion. Blood samples were
collected via a femoral vein during and after the ischemic event. Blood samples were
pretreated and diluted with the appropriate amount of water then injected onto the
graphite furnace atomic absorption spectrometer via an autosampler for the analysis
of magnesium. The linearity of the magnesium concentrations ranged from 0.50 to
3.00 ppb and a detection limit of 0.03 ppb was obtained. The typical intra- and interassay
variation was less than 3%. Magnesium concentrations decreased during
cerebral ischemia and returned to baseline after 60 min of reperfusion.

56
Contribution of the Na+-Mg2+ exchanger on insulin-induced modulation of the
intracellular Mg2+ concentration in rat hearts
Yasushi Tatematsu, Department of Cardiology, Nagoya University Graduate School
of Medicine, Nagoya, Japan; Shinsuke Nakayama, Department of Cell Physiology,
Nagoya University Graduate School of Medicine, Nagoya, Japan; Tetsuya Amano,
Kenji Imai, Manabu Kokubo, Tadayuki Uetani, Takaaki Yamada, Yukihisa
Hamaguchi, Toyoaki Murohara, Department of Cardiology, Nagoya University
Graduate School of Medicine, Nagoya, Japan; Tatsuaki Matsubara, Department of
Internal Medicine, Aichi-Gakuin University, Nagoya, Japan. tatemat@med.nagoyau.ac.jp
In cardiac muscle, numerous intracellular mechanisms are known to be under the
influence of the Mg2+. It is recognized that insulin slowly increases intracellular Mg2+
concentration ([Mg2+]i) in cardiomyocytes. On the other hand, Na+-Mg2+ exchange
is generally considered to play a central role in regulating [Mg2+]i in many types of
cells. However there is little information concerning correlation between insulin action
and Na+-Mg2+ exchange. In the present study, we therefore examined the effects of
amiloride and imipramine on insulin-induced [Mg2+]i rise in isolated rat hearts, using
31P-nuclear magnetic resonance. [Mg2+]i was estimated from the separation of the
chemical shifts of the beta and alpha ATP peaks, using the dissociation constant of
MgATP 87 microM. [Mg2+]i was 1.00±0.08 mM after exposure to insulin (1mU/ml)
for 100 min in the absence of extracellular Mg2+. [Mg2+]i was significantly increased
to 1.21±0.03 mM in the presence of extracellular Mg2+ (1.2mM). When 1mM
amiloride, a poorly selective blocker for many Na+-coupled transporters, was
simultaneously applied with insulin, [Mg2+]i hardly changed for 100 min (in normal
solution containing 1.2mM Mg2+). Furthermore, simultaneous application of 10 mM
imipramine, a selective inhibitor for Na+-Mg2+ exchange, also prevented the insulininduced
[Mg2+]i increase as seen with amiloride. These results suggest that Na+-
Mg2+ exchange may play an important role in the insulin modulation of [Mg2+]i.

57
A study on spontaneously obese rats (Minko rat) with abnormal lipid
metabolism -strength and mineral concentrations in bone
Ryuji Takeda 1,2), Takashi Nakamura 1), Masayo Imanishi 2), Madoka Ishida 2),
Fumiko Yano 3), Takahisa Takeda 2), Mieko Kimura 2), 1) Graduate School of
Medicine, Kyoto University, Kyoto 606-8501 Japan, 2) Takeda Research Institute of
Life Science, Kyoto 600-8813, 3) School of Biology-Oriented Science and
Technology, Kinki University, Wakayama 646-6493. kimura@takedahp.or.jp
Osteoporosis is a major public health problem in many countries, including Japan.
Although bone densitometry is often used as an indicator to evaluate bone fragility,
bone mineral density isn't always reflected in bone strength. It is well known that
there are many factors that affect bone strength. The cholesterol synthetic pathway
may also be important in bone metabolism. In the present study, the relationship
between strength and mineral concentration in bone of spontaneously obese male
and female rats named "Minko rat" with abnormal lipid metabolism is studied. The
bone stiffness was tested by the three-point bending method using a mechanical
testing machine (Model TK-252C: Muromachi-Kikai Co. Ltd., Tokyo Japan), and Mg,
Ca, P, Na, S, K, Zn, Sr and Fe concentrations were determined by inductively
coupled plasma-atomic emission spectrometry (ICP-AES: Perking-Elmer Co. Ltd.,
USA) in femur of 39 weeks old rats. Mechanical studies in bone indicated that the
stiffness of male rats was significantly higher compared to that of female rats. On the
other hand, magnesium, calcium, phosphorus, sodium, potassium, zinc, strontium
and iron concentrations in male rats were significantly lower compared to that of
female rats, while sulfur and potassium concentrations in male rats were significantly
higher compared to that of female rats. These results suggested that bone strength
isn't determined only by mineral concentrations such as magnesium, calcium and
phosphorus. On the other hand, we report that bone strength of "Minko rats" was
significantly higher compared with control rats, but there was no significant difference
of calcium and phosphorus concentrations between the two groups of rats.

58
Modulation of tyrosine kinases and phosphatases by Mg2+ ions in human red
blood cells
Alexander Barbul1, Yehudit Zipser1, Nechama S. Kosower2 and Rafi Korenstein1.
1 Department of Physiology and Pharmacology, and 2 Israel Department of Human
Genetics and Molecular Medicine, Sackler Faculty of Medicine, Tel-Aviv University,
69978 Tel-Aviv, Israel. abarbul@post.tau.ac.il
The anion exchange band 3 protein is the main substrate for phosphorylation by
protein tyrosine kinases (PTKs) in red blood cells (RBC). Dephosphorylation of band
3 is carried out by phosphotyrosine phosphatase (PTP). Under normal conditions,
little if any band 3 phosphotyrosine is present in the RBC, due to higher activity of
PTP versus PTK. We previously showed that band 3 tyrosine phosphorylation was
induced upon elevation of Mg2+ concentration during deoxygenation or Mg2+loading
of intact human RBC. This effect was much more pronounced upon addition
of the PTP inhibitor vanadate. We have now found that Mg2+-induced band 3
phosphorylation is inhibited by the specific Src family kinase inhibitor PP1. Mg2+
leads to activation and autophosphorylation ofSrc-family kinases p53/56Lyn and
p59Fyn, which in turn phosphorylate and activate p72Syk tyrosine kinase. Though all
three kinases appear to be associated with band 3, only p72Syk exhibits a very
efficient direct phosphorylation of the cytoplasmic domain of. On the other hand,
band 3-associated PTP is also found to be activated by Mg2+. Moreover, the addition
of Mg2+ to RBC membranes where PTP was dissociated from \band 3 by Ca2+treatment,
causes re association of PTP and band 3 and dephosphorylation of
band3. Thus, the Mg2+ ion seems to be a significant regulator of phosphotyrosine
levels in human RBC with important consequences for RBC physiological functions
and metabolism.

59
Is determination of Mg pool size useful to assess Mg status
C. Feillet-Coudray, C. Coudray, E. Gueux, A. Mazur and Y. Rayssjguier. INRA, St
Genès Champanelle, France. coudray@clermont.inra.fr
Magnesium (Mg) is a biologically essential mineral and Mg deficiency is known to
lead to severe biochemical and symptomatic disorders. However, a sensitive and
valid marker to assess Mg status in humans is still unavailable. We hypothesised that
determination of exchangeable pools of Mg could be used to estimate tissue Mg
status and turnover. In that aim, studies were conducted in animals and in human. In
animal study, Mg pool sizes were determined in control rats, marginally Mg-deficient
rats and severely Mg-deficient rats. Rats received an intravenous injection of 25Mg,
and the plasma 25Mg disappearance curve was followed for several days.
Tracer/tracee data were analysed with a SAAM II program, using a compartmental
model based on the model of Avioli and Berman (1966). This model considered 3
exchangeable Mg pools with varied rates of turnover (pools 1 and 2. with a relatively
fast turnover. and pool 3). We observed that the size of exchangeable pools of Mg
decreased in proportion to Mg deficiency. We thus conclude that Mg pool size
measurement can constitute a good marker of Mg status in rats. To validate this
concept in human, exploration of Mg pool size was performed in 10 healthy women
before and after 8 weeks of Mg supplementation (366mg Mg/d). All subjects had an
intravenous catheter inserted into the right arm, and 25Mg was perfused over 30 min.
Each subject had also an intravenous catheter inserted into the left arm for blood
sampling at T-30 to T-600 min. On the following days blood sampling was also
performed after an overnight fast (Dl to D7). Total 24-h urinary Mg excretion was
measured for 2 days, on the day before and the day of the isotopic loading test. We
observed that Mg pool size was not affected by the Mg supplementation while urinary
Mg excretion was significantly increased. Full Mg stores prior to Mg supplementation
may explain such results. In conclusion, Mg pool size measurement may be a
promising indictor of Mg status. Further studies are necessary to determine whether
Mg pool size responds to Mg supplementation in Mg-deficient subjects.

60
Effects of reduced magnesium availability and mild oxidative stress on aging
of WI-38 human diploid fibroblasts.
B.Tedesco, S. Fasanella, A.Torsello, A.Cittadini and F.I. Wolf. Institute of General
Pathology and Giovanni XXIII Cancer Research Center, Catholic University School of
Medicine, Rome Italy. fwolf@rm.unicatt.it
Reduced Magnesium (Mg) availability due to reduced intake (diet) or increased loss,
is often found in the elderly. Hypomagnesiemia exacerbates cardiovascular diseases
and diabetes, two major diseases primarily affecting aged people. From a
biochemical viewpoint Mg influences several functions involved in the aging process,
among which proliferation rate and oxidative reactions which can cause important
impairment of
protein, lipids and DNA functions. In particular, DNA damage is involved in the
inhibition of cell proliferation associated with cell senescence. We have shown that
acute and chronic decrease of extra-cellular magnesium reduces proliferation rate of
different cells in culture. We have also shown that a mild chronic oxidative stress
(5mM H2O2) accelerates aging of WI-38 diploid human fibroblasts. In this work we
wanted to investigate the effects of the association of reduced Mg availability and
oxidative stress
on the aging of WI-38 fibroblasts. Results indicate that decreased Mg availability
(from 0.8 mM to 0.4 mM in extracellular medium) slowed down the proliferation rate
of WI-38 cells. The combination of decreased Mg availability and mild chronic
oxidative stress resulted in a reduction of oxidative DNA damage, suggesting that in
this conditions, cell are more resistant to the detrimental effects of oxidative stress.
These results can be explained in the light of the different cellular response to acute
or chronic hypomagnesiemia, and underline the importance of cell/organism
adaptation to sub-physiological Mg availability.

61
Food intake and magnesium intake affect true absorption and endogenous
fecal excretion of magnesium in rats
Hideyuki Ohmori, Tohru Matsui and Hideo Yano. Division of Applied Biosciences,
Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
yanoh@kais.kyoto-u.ac.jp
True Mg absorption and fecal endogenous Mg excretion were studied in rats given
different amounts of Mg (5, 10, 20 mg/d) in different amounts of diets (10 and 20 g/d)
except for 20 mg Mg in 10 g diet. Rats were subcutaneously infused 25Mg by an
osmotic pump and true absorption and fecal endogenous excretion of Mg were
determined after the isotopic enrichments became almost stable in urine and feces.
Feed intake did not affect the amount of Mg absorbed but the amount of Mg
absorption increased with Mg intake irrespective of feed intake. Although Mg
absorption rate (percentage of Mg intake) did not differ between rats given 5 mg Mg
and 10 mg Mg in 20 g diets, rats given 20 mg Mg showed lower absorption rate than
rats given 5 mg Mg. Fecal endogenous excretion of Mg increased with dietary Mg
level and increased with feed intake. Urinary loss of Mg was approximately 4.5 to 7fold
more than the endogenous fecal loss of Mg. These results suggest that the
efficiency of Mg absorption decreases with dietary Mg level and the endogenous Mg
excretion into feces depends on Mg and feed intakes. However, the endogenous
fecal excretion of Mg does not largely contribute to Mg metabolism in rats.

62
Serum magnesium levels and dependency/disability in hospitalised elderly
patients
M Barbagallo, LJ Dominguez, A Ferlisi, A Galioto, A Pineo, C Aglialoro. Gruppo
Italiano di Farmacovigilanza nell'Anziano, Institute of Internal Medicine and
Geriatrics, University of Palermo, Sicily, Italy. mabar@unipa.it
Magnesium deficiency has been linked to age-related chronic conditions such as
hypertension, diabetes and cognitive impairment. Since all these conditions may
favour the development of dependency and disability, we explore the association
between serum magnesium levels and disability-related conditions in the elderly. We
performed a cross-sectional case-control study in the GIFA (Gruppo Italiano di
Farmacovigilanza nell'Anziano) participants, carried out in 81 hospitals throughout
Italy. A total of 5197 elderly subjects (aged 65 to 100 years) with serum magnesium
measurements available were studied. We included in the analysis the following
variables: age, gender, body mass index, blood pressure measurements,
hypertension, diabetes mellitus, laboratory parameters, educational level,
comorbidity, number of drugs used, cognitive impairment (evaluated with Hodkinson
Abbreviated Mental Test), and activity of daily living (ADL) scale. In the univariate
analysis serum magnesium showed a significant trend for negative association with
age, diabetic status, systolic blood pressure, serum calcium, cognitive impairment,
self-sufficiency, atrial fibrillation, osteoporosis and depression (p

63
Absorption and effect of the magnesium content of a mineral water in the
human body
Sándor A. Kiss, Tamás Forster, Ágnes Dongó, 1 Hungarian Magnesium Society,
2 Univ. Szeged, 2 nd Dept. Of Medicine And Cardiology Center, Szeged, Hungary
The kinetics of magnesium absorption in the human body has been investigated by
MAGNESIA mineral water (204 mg Mg/L) on healthy human subjects, aged (23-60
years) in a self-controlled manner. Serum magnesium, calcium, K (potassium) and
Na (sodium) content, blood haemoglobin, erythrocyte and white blood counts as well
as urinary volume and urine magnesium content were evaluated. Subjects drank 1.5
liter MAGNESIA during 30 minutes, blood tests were taken at 0, 2, 6, 24 and 48
hours, and subsequently 1, 2, 3 and 4 weeks. Serum ion quotient was calculated.
Serum magnesium level increased in all cases, though it returned to the individual
normal value after 48 hours. Subjects drank mineral water intensively only on the first
two days, later on they consumed 1-1 glasses of mineral water at a time with a daily
total amount of 1-1.5 liter. Urinary volume and its magnesium content significantly
increased. The increase in the magnesium content of urine was individually different
referring to the divergent level of tissue magnesium deficiency. Our observations are
in accordance with that on plants, i.e. magnesium is absorbed in a short period of
time in all biological systems. MAGNESIA mineral water was provided by
Karlovarske Mineralni Vody a.s., which merits our gratitude.

64
About the misdiagnostics of magnesium deficiency
Dierck-H.Liebscher, Dierck-E.Liebscher. Self-help Organisation on Mineral
Imbalances, Berlin, Germany. dierck-h.liebscher@magnesiumhilfe.de
The experience of our self-help organisation shows that the reason why patients with
magnesium deficiency symptoms do not get magnesium therapy is the badly used
lower limit of the reference values for the serum concentration of the normal
population. This limit is too low for a patient with symptoms so that the prevalence
and importance of this disease is not taken into consideration sufficiently. The lower
reference limit of the normal population is erroneously regarded as a diagnostic
criterion to exclude the deficiency when more Mg is found, while it only indicates
exclusion of normality when less is found. It is a famous error in statistics to use the
confidence limits of the normal population as exclusion limits for the affected. We
demonstrate a calculation to estimate the number of patients, who are not correctly
diagnosed, as a function of the lower limit for the serum value based on data of
v.Ehrlich (1997). Among 3894 patients, 366 had symptoms belonging to a clinically
relevant magnesium deficiency but only 37 had a serum value of lower than the
presently used limit of 0.7 mM Mg.
Our conclusions are
- When there is a chance for causal therapy - and magnesium therapy is - then this
therapy must be the first choice.
- When Mg-deficiency symptoms are diagnosed, the serum value must be checked.
- Patients with symptoms and serum values lower than 0.9 mM Mg have to be
involved as suspected Mg-deficient patients.
- When Mg-deficiency symptoms are diagnosed, substitution must start with more
than 600 mg Mg per day.

65
Magnesium in asthma attack
Dr. G. Sur, Prof. Dr. N. Miu, Dr. Lucia Burac, Dr. Oana Maftei. Pediatric Clinic II,
University Of Medicine And Pharmacy, Cluj-Napoca, Romania.
oana_maftei@yahoo.com
The purpose of this study was to analyze the most efficient therapy for patients with
bronchial asthma and to establish the role of magnesium in the treatment of
bronchial asthma crisis. Magnesium has been used as an adjuvant drug in bronchial
asthma crisis therapy, along with the usual therapies established by Bethesda and
Gina programmers. Based on previous studies in the Clinic of Pediatrics II before
1989, the authors concluded that i.v. administration of Mg with specific medication of
crisis reduces the period of crisis and the hospitalization. We found that earlier i.v.
administration of Mg in moderate and severe crisis of asthma is beneficial both for
the rapidity of response and for the improvement of the symptomology. In our study,
we noticed that oral administration of Mg has no efficacy, no matter what the dose.
For efficiency in asthma is necessary to administer i.v. doses: 50 mg/kgc in 20
minutes to 2 hours.
Mg is the 4 th most abundant cation in the organism and the second most abundant
intracellular cation. It coordinates the activity of many enzymes of cellular
metabolism and is an important cofactor in oxidative metabolism. The excess of Mg
in smooth muscles induces calcium decrease, which is necessary for myosin
phosphorylation. Mg is a competitor of calcium for the linked tissue and it may
increase the activity of ATP in cellular membranes. Excess intracellular Mg favors the
entrance of calcium in endoplasmic reticulum and that's why it doesn't induce
calmodulin activation and the preformed and neoformed products of mastocyte and
basophile. Today it is well known that Mg is a calcium antagonist so that it
decreases the level of calcium in smooth muscles and produces relaxation. It also
inhibits acetylcholine in the neuromuscular junction; inhibits histamine; directly
inhibits contraction of smooth muscle; produces sedation; activates adenylate
cyclase, which increases ATP, which induces cyclic AMP; and is bronchodilatory. We
studied 58 patients hospitalized in Pediatrics Clinic II for the last 5 years, with
moderate and severe crisis. We administered to these patients Mg 20-50 mg/kg in 20
minutes. We followed the decrease of crisis severity, the improvement of
symptomology and the period of hospitalization.
In this study group we found:
1. In bronchial asthma crisis therapy, the magnesium proved its efficacy,
administered only when it is earlier and in efficient doses of 50-75 mg/kg in 20
minutes to 2 hours.
2. The magnesium reduces the crises, significantly reduces the period of
hospitalization and improved the ventilator parameters.
3. If we can't control the level of magnesium in blood serum, it is better to not
exceed 75 mg/kg because it can cause adverse effects.
4. We mention that it is very important to administer the magnesium at the same
time as the specific medication for different severity degrees of asthma attacks.
We conclude that Mg causes: diminution of crisis period; quick improvement of
ventilation parameters; and diminution of hospitalization.

66
Experimentally induced prolonged magnesium deficiency causes osteoporosis
in the rat
Stendig-Lindberg G, Koeller W, Bauer A, Rob PM. Dept Physiol And Pharmacol,
Sackler Faculty Of Med, Tel-Aviv University, Ramat Aviv, Israel And Dept Of
Orthopedics And Dept Of Dialysis, Lubeck University Hospital, Lubeck, Germany.
Lindberg@Post.Tau.Ac.Il
BACKGROUND. Prolonged daily oral magnesium (Mg), given as the only treatment
in postmenopausal osteoporosis, caused significant increase of BD.
OBJECTIVES. In order to obtain definitive evidence of causality of magnesium
deficiency in the etiology of osteoporosis, we examined throughout one year, rats
given daily a Mg adequate diet (2000 ppm), Group A, to compare with rats given
daily a Mg deficient diet (200 ppm), Group B.
METHODS. Sixteen Sprague-Dawley female rats, mean wt.110, SD 23 g, were
randomly divided into group A and B. Urine samples were collected every three
months and blood, at the end of the trial. After the animals were sacrificed, L3 - L5
vertebrae and the femural regions were examined for bone density (BD) using dual
energy X-ray absorptiometry. The femural bones were examined for bone fragility,
and the tibiae by histomorphometry.
RESULTS. The mean BD of L3-L5 vertebral bones and of the femural bones was
significantly higher in group A than in group B (p =0.035 and p = 0.045, 1 tail,
respectively). The bending stiffness of the femur in Group B was significantly higher
than that in group A (p = 0.024) and the force needed to break the bone was higher
in group A, than in group B. Histomorphometry showed in group B a diminution of the
trabecular bone volume, in relation to the tissue volume (BV/TV), an increase in
the degree of the trabecular interconnections (TBPf) and focal osteoporosis of the
metaphyseal spongious bone.
CONCLUSIONS. Rats kept one year on a Mg deficient diet developed osteoporosis.

67
Modifications of the plasmatic and salivary magnesium concentrations and of
other bivalent cations in patients with suppurations of the oro-maxilar area
1 Nechifor Mihai, 2 Gradinaru Irina, 3 Mîndreci Ioan, 2 Tatarciuc Monica, 4
Gogalniceanu Dan, University of Medicine and Pharmacy "Gr.T.Popa" Iasi, Romania
1 Clinical Pharmacology Department, 2 Complex Oral Rehabilitation Department,
3 Biophysics Department, 4 Oro-Maxilo-Facial Surgery Department.
nechifor@umfiasi.ro
We surveyed the plasmatic and salivary concentrations for Mg2+, Ca2+, Cu2+, Zn2+
for adults patients, males and females, presenting suppurations of the maxilo-facial
area. The registrated data were compared to a reference group, represented by
healthy patients with the same age, sex and dietary habits. We noted that the
patients with suppurations presented a significantly higher concentration of the
Mg2+/Ca2+ ratio in comparison to the healthy patients. The plasmatic concentration
for Mg2+, Ca2+ was not significantly different between the two groups. The plasmatic
concentration for Zn2+ is significantly smaller for the patients with suppurations
(1,06±0,12 mg/l) in comparison to the reference group (1,4±0,14 mg/l), p

68
Lyme disease and magnesium deficiency
V. CRISTEA - Department of Immunopathology, Medical Clinic III, "Iuliu Hatieganu"
University of Medicine and Pharmacy, MONICA CRIAN - Department of Immunology,
"Ion Chiricu" Oncological Institute, Cluj-Napoca, Romania
V. CRIAN - ITEM-Paneuro Group. vlaicu@mail.dntcj.ro
During the period April 2001 - January 2003, we had under observation two cases, in
which the presence of both IgM and IgG antibodies to Borrelia burgdorferi was
serologically confirmed at high titers. In both cases, clinical manifestations were
similar: shivering, fever, headache, articular and right hypochondrium pain, and
objectively - tachycardia and erythema migrans - these elements being important for
the formulation of Lyme disease suspicion. Humoral tests showed: significantly
increased ESR, leukocytosis with PMN predominance, intensely positive PCR (for B.
Burgdorferi DNA) and significant magnesium deficiency (1.20 mEq/L, 1.33 mEq/L,
respectively). A large spectrum of antibiotics with both oral and parenteral
administration has been so far used in the treatment of Lyme borreliosis. Among the
most frequently used are tetracyclines, betalactamides and cephalosporins. The
decision to initiate antibiotic therapy can be difficult because in the majority of the
cases acute infection is self-limited. Asymptomatic patients, in whom laboratory
examinations sustain the diagnosis of Lyme disease, should be treated in order to
prevent infection dissemination. Since in the first case antibiotic therapy alone did not
lead to the expected results, magnesium derivatives were also associated. In both
cases, following combined therapy, symptomatology significantly improved at 14
days, and laboratory examinations were restored to normal values after 6-8 weeks -
disappearance of IgM to B. Burgdorferi and significantly increased magnesemia
(1.74 mEq/L, 1.72 mEq/L, respectively) We believe that in certain diseases, Mg
deficiency can cause a decrease in immune response. The appearance of
recurrences, which are frequently reported in the literature, in spite of adequate
antibiotic therapy, could represent an argument for this. This is why the use of Mg
derivatives in therapy can represent an immunostimulating factor. The peculiarities of
the cases are the following:
1. Patients had in addition to fever, articular pain and erythema migrans, Mg
deficiency
2. The supplementation of therapy with Mg derivatives had an immediate beneficial
effect that was maintained in time.
As a conclusion at this stage, we consider that in the acute phase of Lyme borreliosis
there is a significant Mg consumption and the introduction in therapy of such
preparations is recommended and beneficial.

69
Effect of magnesium on essential oil formation of genetically transformed and
non-transformed chamomile cultures
Szokee, É., Máday, E., Kiss, A.S. and Lemberkovics, É. Department Of
Pharmacognosy, Semmelweis University Of Medicine, H – 1085 Budapest, ÜllI Út
26, Hungary. Szokee@Drog.Sote.Hu
The importance of chamomile (Chamomilla recutita Rausch.) is widely known in
medicine. The largest group of its effective substances form the essential oil
(chamazulene, a-bisabolol, trans-b-farnesene, spathulenol, cis/trans-en-indicycloethers).
The increasing need for high quality drugs cannot be provided by
plant collection in the wilderness. To preserve the genome of Szabadkígyós wild type
having high (-)-a-bisabolol content, we used biotechnological methods. The roots of
organised culture contained b-eudesmol, wich was firstly identified from the intact
roots by us. Our gas-chromatographical and mass-spectroscopical studies showed
that sterile chamomile cultures generated the most important terpenoid and polyin
compounds characteristic of the intact plant. We identified berkheyaradulene,
geranyl-isovalerate and cedrol as new components in these cultures. Magnesium
(370 and 740 mg/l MgSO4) has positive effect on the growth of organized cultures
and also on the quality and quantity of essential oil production. Another possible
source of such variants is available by the genetic transformation of organized
cultures by infection with Agrobacterium rhisogenes. Adopting this method we
cultivated chamomile infected by A4-Y clone and investigated the essential oil
production by hairy root cultures cultivated on solid and liquid MS respectively B-5
media. The main component of the essential oil of hairy root cultures was trans-bfarnesene.
We identified a-selinene as new component in these hairy roots. We
studied the growth rate of A4-Y clone on media mentioned above containing different
MgSO4 concentrations (0; 185; 370 and 740 mg/l). The cultures were growing the
most intensively in medium containing 740 mg/l. MgSO4. Essential oil content was
compared obtaining from hairy root cultures of different magnesium containing media
and measured by GC and GC-MS methods. Magnesium has similar effect on hairy
roots as on organized cultures.

70
Magnesium in animal nutrition
Katalin Kovacsne Gaal - Orsolya Safar - Laszlo Gulyas - Petronella Stadler.
University of West – Hungary, Faculty of Agricultural and Food Sciences, Institute of
Animal Breeding, 9200 Mosonmagyarovar Var 4, Hungary. safors@freemail.hu
Performance and reproduction of animals depends to a great extend on their energy
supply. Magnesium plays an important role in the metabolic energy supply.
Magnesium supplements remarkably improved the digestibility of feed. Magnesium
citrate and magnesium oxide improved the exploitation coefficients by 0,30% and
0,50% respectively and the improvement was 4,71% in respect of crude protein. In
connection to its biological value, the improvement has been more significant
because the feed with 0,15% magnesium citrate was better utilised (83,18%) than
the control feed (70%). Magnesium was fed to gilts from the start of the heat- and
ovulation sychronisation until the day of the first insemination (25 days). The dietary
magnesium citrate surplus in the doses of 15 vs. 30mg/kg/body mass/day improved
the conception rate by 11,3 vs. 14,7% and the litter size by 11,9 vs. 12,9%
respectively. In case of cows, a supplementation of the feed with 20g Bolifor MGP
(animal) day improved the reproduction output and shortened the service period by
7,93%. In the case of boars the amount of ejaculation grew by 1,48% as a minimum
and 30,9% as a maximum, but the quality of semen produced in both experiments
remained unchanged. In further experiments the feed ration of broiler chickens was
supplemented either with 0,2% magnesium citrate or 0,2% MgO through a 48-day
experiment. The average live weight of the control and experimental groups (Mg
citrate and MgO) were 1,27; 1,32 and 1,38 respectively. We supplemented the diet of
Tetra SL laying hens either with 0,4% Mg citrate or with 0,4% MgO. Mg citrate
supplementation increased the egg production by 9,68% compared to the nonsupplemented
control group. We aimed to investigate the effects of magnesium
supplementation on the parameters of producing eggs for breeding, hatching yields
and embryonic development, the mineral content of egg shell and intestines of chick
embryo. Three magnesium rations were applied in our experiments (Mg1 300, Mg2
400, Mg3 500mg/Mg/day) in the form of the product HAMAG LP. Coinciding with the
levels of the Mg supplementation the growth of the embryo showed a linear increase
at all the four times of dissection. Furthermore, we investigated the weight and Ca, P
and Mg content of egg shell and that of the viscera of embryo at the age of 14, 16, 18
and 20 days together with brain, liver, heart and tibia. Expressing the size of internal
organs in the proportion to the embryo weight we can observe that in the period of
14-18 days the liver of the embryo gained on weight at the same rate as the level of
magnesium supplementation was increased. Compared to the control no consecutive
gain on weight of the other viscera could have been observed in the test groups.
Feeding magnesium above the requirement lowered the calcium content in the egg
shell. Our investigation showed that the calcium content of the shell lowered as the
rate of magnesium feeding increased in the progress of hatching however but the
magnesium content of the shell increased. Therefore the reduction of calcium
content and the increase of magnesium content can be explained by the classical
antagonism between the two elements. The content of phosphorous, calcium and
magnesium of the viscera organs of the embryo did not show any changes following
the magnesium feeding either days of the dissection. As a summary we can say that
magnesium feeding benefits the quality of breeding eggs, improves hatching yield
but magnesium does not incorporate into the viscera of the embryo although brain
and liver may have a function of storing organs for magnesium intake above the
requirement.

71
Mg-content of Rhizobium nodules in different plants and the importance of Mg
in N2-fixation of nodules
Kiss S.A. 1 , Stefanovits-Bányai É. 2 , Takács-Hájos M. 3 1 Hungarin Magnesium Society,
H-6726 Szeged, F_ str. 73 A/2 Hungary; 2 Szent Istvan University Faculty of Food
Science Department of Applied Chemistry H-1118 Budapest, Villányi str. 29-31.
Hungary ebanyai@omega.hu; 3 Tessedik S. College Agricultural Water and
Environment Management Department, Szarvas, Hungary. hajos.maria@mvk.tsf.hu
Rhizobium bacteria induce nodules /tumours/ in roots of leguminous crops. These
nodules fix N2 from the atmosphere. The N2 fixed in this way is used partly to build up
their own bodies and partly to be transported to the host plant/symbiosis. The
bacteria need materials from the host plant /ATP/ which supply energy. Rhizobium
strains differ in their N2-fixing ability have higher Mg- requirement. Both the enzyme
synthesis and the proteins necessary to form nodules are produced in ribosomes.
This process also needs higher Mg quantities. As the enzymes taking part in N2fixation
have a very short life time /30 sec/ their reproduction needs increased
enzyme synthesis. Trials were carried on in sterile perlite where the two peas
species were growing hidroponically, and in two different soil types (brown forest soil
and meadow silt soil) in field where different leguminous plants were chosen for test
plants: lupin, soybean, broad-bean, lentil and bean inoculated by Rhizobium bacteria.
The Mg-content of the nutrient solution was higher than that of the control, but in the
soil tests 1 % Mg leaf fertilization was applied after bacterial inoculation. Number,
weight and Mg-content of nodules were evaluated using an AAS method compared
with the thick and thin /hair/ roots of the given plant. Our results clearly shown Mg
supplying increased the height of the plants. Proper Mg supply increased the number
and weight of nodules considerably, which preconditions N2-fixation and higher yield
of the host plant. Mg nutrition /treatment/ increased the number and Mg-content of
Rhizobium nodules resulting in increased N2-fixation and yield.