HEALTH AND NUTRITION NEWSNo. 7
March 15, 2004 volume 2 number 4

Table of Contents
PREFATORY NOTE
Lifestyle–related diseases and National Institute of Health and Nutrition…………....…….………....…….……………....…………....…………....……....………Nobuaki KUZUYACURRENT RESEARCH PROJECTS
Checking the data for the National Health and Nutrition Survey in Japan……………………………………………………………………………………………...…Katsushi YOSHITATHOUGHTS ON HEALTH AND NUTRITION RESEARCH
A Thought for Taking Diet ……………………………….….……………...………..…Kazuhiko YAMADARESEARCH FINDINGS
Negative balance of calcium and magnesium under relatively low sodium intake in humans.……….. ………..…... ………..…... ………..…... ………..…... ………..…...…... Mamoru NISHIMUTA
Regulatory sequence elements of GLUT4 gene expression in adipose tissues………………………..……………………..……………………..…………………..…………Shinji MIURA
How much exercise is required to reduce blood pressure in essential hypertension?
………………………..……………………..……………………..…………………..………Kazuko TAKATA
Developing a functional structured lipid efficient for the prevention of life-style related disease………………………..……………………..……………………..…………………..………Junichi NAGATA© National Institute of Health and Nutrition

Prefatory Note
Lifestyle –related diseases and the National Institute of Health and Nutrition
Dr. Kuzuya Nobuaki
Director of Clinical Laboratory,
Department of Endocrinology and Metabolism, International Medical Center of Japan(adipocytokines), which causes insulin resistance
and/or promotes impaired glucose tolerance, highblood pressure, lipid metabolism disorder, andarteriosclerosis. Also at a molecular level, thefunction of enzymes such as PPARγ, PGC-1,
SREBP-1c, as well as UCPs which reduces thelipid storage in tissues, GLUT4, and AMP kinase all
gradually has become clear. National Institute of
Health and Nutrition publishes many interestingfindings which relates to these nutritional
substances.

Lifestyle–relateddiseaseisdefinedas“syndromes related to one’s
lifestyle such as diet, physicalexercise, rest, smoking anddrinking with its onset anddevelopment”.
The
population of lifestyle-relateddiseases such as diabetes, high blood pressure,
hyperlipidemia, and obesity has significantly
increased in recent days. Each disease above
can be a risk factor for developing arteriosclerosis.
Also, a patient may be affected with more than one
lifestyle-related disease.
Blindness due todiabetic
retinopathy
andpatients
underhemodialysis by diabetic renopathy both increased
Cerebral infarction and ischemic heart disease,which are arteriosclerotic ivents, increased in theelderly. This increase in lifestyle-related diseases
not only occupies the leading causes of death inJapan, but also leads to the increase in elderly whoneed special care.., Causes of diseases arecomplex, but are mainly divided into hereditary andenvironmental factors. However, it is said that the
rapid change was brought to us by the change ofsocial environment, westernized life style, andrapidly aging population.

Diet and exercise are the fundamental
treatments for lifestyle-related diseases. We needto give our patients a clear explanation about themerit of the diet and exercise and encourage themto change their life style. I think it is important tofind out how and what kind of life style factors are
related to cause which disease, at molecular, atcellular, at organ, and at individual levels.
Internationally, WHO points out that
diabetic population will increase 1.9 times within 22
yeas in developing countries in Asia. Yet thesecountries have not fully developed a dietitiansystem and sometimes nutritional and dietetic
policies are inadequate.
I expect National
Institute of Health and Nutrition to collaborate withthese countries in develop their health promotion
measures. I believe that this is what National
Institute of Health and Nutrition is expectednationally, as well as from a global level, in addition
to health promotion, prevention and medical
solution of diseases.

As you all know, lifestyle-related diseaseis related to obesity. It is speculated that insulin
resistance, which is caused by obesity, promotes
the lifestyle-related diseases.
Adipose cells
produceandsecretemany
substances2Current Research Projects
Checking the data for the National Health and Nutrition Survey in Japan
Dr. Katsushi Yoshita (Head of Data Management for National Health and Nutrition Survey)

What do you imagine when you hear“checking the data”? Many of you would think ofsomething like counting the check marks or
numbers of multiple choices from the question
sheets. However, “checking the data” for the
National Health and Nutrition Survey in Japan is
not that simple. The survey, which is based on
the Health Promotion Law, is quite unique incomparison to other administrative surveys. We
request over 5,000 households, to record the kindof food and their quantity the individual family
members eat, and with the details such as how
much soy sauce is used by a person, and so on.
In addition, we hold the medical checkup site near
the selected areas to get the survey subjects’
data on blood tests, blood pressure, and about
drug use, etc… This complicated survey isstated as the mission of our institute in the law.
We are required to “check” the complex and vast
data of the survey.

simple but essential process, we start working on
the complicated records of diet, and there we findsimple mistakes, such as ages and sexes.
When we go through more than 10,000 survey
subjects’ data, we correct uncountable minor
mistakes.
After these manual checking, we put
these data into a computer. Step by step, thesurvey data are cleaned by a computer programand by registered dietitians, then finally, we canstart the calculations. At last, about 12 months
later the survey is done, the ministry of Health,
Labor and Welfare announces “the summary of
the National Health and Nutrition Survey inJapan” as you would read and hear on the news.
Because we handle huge records, therelated process for checking, inputting, andtallying up is also enormous. To reduce thelabor and increase the accuracy, we havedeveloped the computer system which connects
health care centers and NIHN. Thanks to thissystem, we now reduce much time comparing toseveral years ago. Our team of dietitians keep
working hard to step up the system. I would beglad if you remember our staff’s such hard workwhen you have a chance to see the result of the
National Health and Nutrition Survey in Japan.

Since the survey is annually conducted
in November, at the Christmas season we receive
the data sheets sent from health care centers of
300 areas all over Japan. Then a new year
comes, we start the checking. First we checkbunches of the papers if all are set in orderbecause there are several varieties of records
and survey sheets for each survey subject, andeven one sheet can not be missed. After this3

Thoughts on Health and Nutrition Research
A Thought for Taking Diet
Dr. Kazuhiko Yamada (Director of Division of Applied Food Research)
It is wonderful if you can move yourmuscles without pain, have fulfilled feelings,
being secured socially, and enjoy life withflexibility without being affected by inconvenientchanges in environment.
World HealthOrganization defines health as “a state ofcomplete physical, mental and social well-being
and not merely the absence of disease or
infirmity” in the preamble of its charter. We, asJapanese, can take phrase for granted, but not inmany countries and areas in the world.

the fibers which is clear to have biological
function by scientific evidence. Japan takes
dietary fiber as a sum of food ingredient involving
animal derived which is undigested by humandigestive enzymes, and we undertake theanalysis according to this definition. This kind of
discussion influences standards of food intake,nutritional ingredients labelling, Standard Tablesof Food Composition, and National Health andNutrition Survey.
Now I think we need tounderstand the international points of view.

I received an e-mail from a friend who is
working in Afghanistan as a JICA (Japan
International Cooperation Agency) officer. In theletter he wrote; I wish you a happy new year!
And no war on the Earth! Here we have anomen of reconstruction from the annihilation
brought by the 20 years of war and 4 years ofdrought. I will keep working to help the recoveryof Afghani farm villages this year as well. Every
morning I have nan bread, eggs (boiled oromelet), cheese, and a cup of coffee or tea at thehotel. Tea in Afghanistan is usually green tea
but it is a little different from a Japanese one.
Lunch is served at the office and I have it with mycolleagues.
Menu is boiled vegetables on
steamed rice and nan bread. Yesterday I had
potatoes, cauliflower, beans, and carrot andgreen peas for today. When I told this to aperson at JICA office he said ‘That’s all
carbohydrate!’, but I do not care. Occasionally I
am invited at the restaurant dinner with specialists.
In Afghanistan, there are Afghan, Chinese, Italian,Western, and Indian cuisines. There is jiao-zi socalled ‘mantou’ which is eaten with yoghurt in
Afghanistan. That reminds me of ‘momo’, the
Nepalese jiao-zi. Jiao-zi must come down along
the silk road from China.” I felt the history andculture of food from his mail.

In my graduate student days, I saw apicture at the front page of a text book of nutritionin the library. In the picture there were 26
smiling children from 26 different countries in theirown traditional costumes.
With the caption
which reads “These children have different
breakfasts in different cultures, but they are all
healthy and growing well.
The essence ofnutrition is to find out what is in common.” WhenI attended the Asian Congress of Nutrition, in aworkshop on diet in different countries at thisconference, one speaker responded to a question
asking back, “Do you mean change our diet?”
Then the person who asked said, “No, no I mean
modify the diet.” followed by a kind of softened
atmosphere. I can imagine there are varieties ofways to promote health through healthy eating,under different cultural conditions.

I participate in some committees ofCodex Alimentarius as a part of my job. As thisisaninternational
meetingforfoodstandardization, here we discuss on such issues,
for instance, “Emphasis of nutritional functions offood” or “International unification of energy
conversion on food labels”. Dietary fiber, for an
example, is a hot topic because its definition andanalysis methods are not universal.
Somecountries insist dietary fiber should be ofplant-origin, on the other hand, some claim to add

4Research Findings
Negative Balance of Calcium and Magnesium under Relatively Low Sodium Intake in
HumansDr. Mamoru Nishimuta (Chief of Laboratory of Trace Element Metabolism, Division of Human Nutrition)
The sweat Na was relatively lower, but
the sweat Ca and Mg were obviously higher
during the previous experiment while taking a
relatively low mineral diet with dietary Na of 100
mmol/d (or sodium chloride 6 g/d).

Samples were wet- ashed by hot plates
using nitric acid and hydrogen peroxide, if
necessary, and the minerals (Na, K, Ca, and Mg)
were analyzed by an atomic absorption
spectrophotometer.

To explain these results, one assumption
has been proposed. When Na reaches aninsufficient level in the body, it is eluted from thebone where it is stored to compensate for any
shortage. Then excess Ca and Mg may also be
eluted along with the Na, and thus flow into theblood stream, and thereby inevitably cause areduction of intestinal absorption as well as anincrease in the urine excretion of these minerals.
The aim of this study was to measure the balanceof minerals (Na, K, Ca, and Mg) as well as toevaluate the hormones affecting the Na
metabolism in humans under a low Na intake.

Diet contained a relatively low Na (100mmol/d or 2.2g/d) with adequate Ca (20 mmol/dor 800 mg/d) and Mg (12 mmol/d or 280 mg/d).
Both the plasma renin activity (PRA) andaldosterone level were above the reference
ranges throughout the experiment, which impliedthat the subjects were Na deficient. However, theurine Na excretion was about the same as thatingested, while there was no substantial reductionof sweat Na concentration observed during
moderate physical exercise (13.2 ± 2.6 mmol/L)(mean ± SD). On the other hand, the urine Ca
and Mg levels were high, but the apparent
absorption of Ca and Mg was moderate (21 ± 5%,
34 ± 4%, respectively), which resulted in anegative balance of these two elements. It seems
that the stored Na in the bone is eluted so as tocompensate for the low dietary Na intake, while
any excess Ca and Mg also inevitably flows into
the blood stream with Na, which inhibited theintestinal absorption of both Ca and Mg andaccelerates their excretion in urine.

A 17-d metabolic study, including two
successive balance sessions of 5 d, wasdesigned. Six female students took part in thisstudy after giving their written informed consent
and also receiving a full explanation of the
purpose and methods of this experiment.
During the period, starting from 6:00 a.m.
on day 2 and ending at 8:30 a.m. on day 17, urine
specimens were collected, and the 24-h urine
excretion of minerals (Na, K, Ca, and Mg) was
measured. On the morning of days 4, 9, and 14,
the fasting morning blood was sampled, and thesubjects also orally took a coloring marker for
their feces (Carmine 0.5g) just before breakfast.

Reference: Kodama N, Nishimuta M, Suzuki K.
Negative balance of calcium and magnesium
under relatively low sodium intake in humans.
J Nutr Sci Vitaminol. 2003;49(3):201-9.

Three subjects pedaled a bicycle
ergometer for 60 min once a day at the subject’s
selected intensity (1.25-1.5 kp, 50-60 rpm) during
the first balance session. The other three subjects
did the same during the second balance session.
Arm sweat during exercise was collected after
cleaning the skin surface.
The diets supplied during the experimentconsisted of a five-day rotating menu and met all
normal dietary allowances in Japan as calculatedby the food tables. Fecal specimens werecollected throughout the experiment and wereseparated into those originating in the diet during
the first and second balance periods according to
the ingested coloring marker appearing in thefeces.
5

Research Findings
Regulatory sequence elements of GLUT4 gene expression in adipose tissues
Dr. Shinji Miura (Senior Researcher, Division of Clinical Nutrition)
Glucose transporter GLUT4 is mainly
expressed in the muscles and adipose tissues,and takes an important role to maintain blood
glucose level. Carbohydrates from food aredecomposed into glucose and carried to blood.
When blood sugar(=glucose) level gets higher,
the pancreas secrete insulin which works onmuscles and fat to activate GLUT4 to enhance
glucose uptake in those tissues, so that the
glucose level lowers. Under insulin resistantconditions, the effect of insulin in muscles islimited. Therefore, blood glucose level would noteasily come down.

the transgenic GLUT4 gene into fertilized mice
eggs, so that in some cases mice with transgenic
genes were born. By examining the expression oftransgenic GLUT4 in adipose tissue in these mice,
we have found out what length of the region in thegene is needed for GLUT4 expression in adipose
tissue.
Consequently, now we have succeeded
to narrow the gene areas between adipose tissue
specific elements (ASE) and high-fat responsive
elements (HFRE).
We have found thenucleoprotein X which bind to ASE and its DNA
sequences as well.

It is known that amount of GLUT4 in
adipose tissue is reduced, when fatty meals(Westernized diet) is taken for a long period.
Since the uptake of glucose mainly occurs inmuscles, the insulin resistance in muscles has
been the center of interest. However, as micedesigned without GLUT4 in their adipose also
showed the insulin resistance, it is now
speculated that the reduction in the developmentof GLUT4 in adipose tissue, also relates
significantly to the onset of diabetes.

We are planning to ascertain DNA
sequences of ASE and HFRE, then identify thetranscription
factorsfor
controllingthedevelopment of nucleoprotein. If it comes clearthat these transcription factors controls thedevelopment of GLUT4 in adipose, it may help tocreate new treatments for diabetes.
Reference: Miura S, Tsunoda N, Ikeda S, Kai Y,
Ono M, Maruyama K, Takahashi M, Mochida K,
Matsuda J, Lane MD, Ezaki O. Regulatory
sequence elements of mouse GLUT4 gene
expression in adipose tissues. Biochem. Biophys.
Res. Commun. 2003; 312:277-284.

In our laboratory, we have studied genes
which control the amount of GLUT4 expression.
We have developed transgenic GLUT4 genes
containing GLUT4 coded genes and different
lengths of promoter sequences. Then we put

6Research Findings
How much exercise is required to reduce blood pressure in essential
hypertension?
Dr. Kazuko Takata (Senior Researcher, Division of Health Promotion and Exercise)
It is well known that exercises helps toimprove mild high blood pressure. Then, howmuch exercise reduces how much bloodpressure? This survey was undertaken in order
to discuss and figure out the appropriate
momentum and frequency of exercise.

over 120mins) in a week. The average systolicblood pressure was the same in each group
before the training period.
After 8 weeks,systolic and diastolic pressure of the control
group did not change.
However, all four
exercise groups reduced their blood pressure.
Compared to the 30~60mins group, systolic bloodpressure was improved quite much in 61~90mins
group, but we did not find outstanding difference
between 61~90, 91~120, and over 120 groups.

This study was done to examine theefficiency of exercise against mild chronic
diseases, in which 1,425 subjects worked out for
8 weeks under the supervision of experts toobserve the change of the disease condition.
There were 450 subjects with high blood pressure(categorized in Stage1 and 2). We analyzed 207
subjects’ data with the exception of those who
were under medical treatment of high bloodpressure and/or coronary artery diseases, or had
exercise habits. Before and after the 8-weektraining period, height, weight, blood pressure,
blood tests and dietary intakes and physicalfitness survey were taken. Those who were in
the exercise group had worked out under expert
guidance at a cooperated sports gym. The
exercise was the combination of warm-up,
stretching, aerobic and muscular exercise. The
intensity of aerobic exercise was set to the 50% of
maximal oxygen uptake. On the other hand, thecontrol group kept the daily physical activity
without any special work outs.

Also we compared the frequency of thework out as 1~2times, 3~4times, over 5times per
week, and checked the change in blood pressure.
As a result, those who worked out over 61mins in
total in a week, reduced his/her blood pressure
despite the frequency.
Now it is clear that;1)
If a mild high blood pressure person
works out over 61mins per a week, he/she will
reduce his/her pressure more than 10mmHg in 8
weeks.
2)
The efficiency of the exercise does notdiffer according to the frequency if one keeps over
61mins exercise in total in a week.
Reference: Ishikawa-Takata K, Ohta T, Tanaka H.
How much exercise is required to reduce bloodpressureinessential
hypertensives:adose-response study. Am J Hypertens.
2003 ;16(8):629-33.

Then the exercise group was divided
into four groups according to their amount of workout time (30~60mins, 61~90mins, 91~120mins,

7

Research Findings
Developing a functional structured lipid efficient for the prevention of life-style
related disease
Dr. Junichi Nagata (Chief of Laboratory of Nutritional Assessment of Food Components, Division ofFood Science)
Although you have not heard a term
“Structured Lipid”, it becomes more and morefamiliar to our daily diet. In general, lipids are
structured by 3 fatty acids attaching to a glycerol.
The fatty acids attached to the sn-1, 3 positions of
the glycerol are hydrolyzed by digestive enzymes,
and a mono-glycerol and two fatty acids areformed. “Structured Lipid” is the physiologicallyadvantaged lipid using the biological feature oflipids, to take in this multi-functioning acideffectively into the body.

feeding of highly purified L-M-L types could
effectively improve serum and liver lipid profiles
and that M-L-M types may be a preferable
substrate for the pancreas and contribute energy
supply in rats. Also it shows that M-L-M typestructured lipids show high affinity for digestive
enzymes, and have remarkable function onenergy substrate and supply of essential fatty
acid.
The biological efficiency of lipids is thought
to be indicated that there are some possibilities tobe affected by its basic structure of triglyceride,
not only by the efficiency brought by fatty acids.
Thus, we expect to develop various combinationsof functional fatty acids for the purpose of
prevention and improvement of life-style relateddisease.

At the present time, there are mainly two
types of structured lipids in Japanese market.
One is made to combine a random fatty acid at a
random position of glycerol to bring out thefunctions of the fatty acid most effectively. The
other is processed in a unique form for absorption
by contriving numbers or combining positions offatty acids. In the market, you can finddi-acylglycerol as edible lipids or synthesized oilfrom medium chain fatty acid and edible lipids, as
examples of structured lipids.

Reference: Nagata J, Kasai M, Watanabe S,
Ikeda I, Saito M. Effects of highly purified
structured lipids containing medium-chain fatty
acids and linoleic acid on lipid profiles in rats.
Biosci Biotechnol Biochem. 2003 ;67(9):193743.

In the present study, we have synthesized
four new structured lipids by using the sn-1, 3
specific lipase. Each lipid includes medium
chain fat acid C8:0 or C10:0 and functionallyunique with combination of linoleic acid. Then
we started the study in the purpose of examining
the effects of highly purified structured lipids onserum and liver lipid profiles in rats. We also
investigated in vitro hydrolysis of lipid emulsions
by porcine pancreas.
M-L-M types of structured lipid (medium
chain fat acid at sn-1, 3 positions and linoleic at
sn-2 position of glycerol) were 2 to 3 times higher
than those of L-M-L types (linoleic at sn-1, 3
positions and medium chain at sn-2 position).
The diet containing structured lipids or corn oil
was administrated to rats for 4 weeks. There
were no significant differences in growth and foodefficiency.
Serum cholesterol levels weresignificantly lower in L-8-L, 10-L-10 and L-10-Lgroups than in the corn oil groups. Serumtriglyceride levels were significantly lower in rats
fed L-M-L types than those in other groups.
Serum non-esterified fatty acid (NEFA) andbeta-hydroxybutylate levels were significantly
higher in rats fed M-L-M types than those of theother groups. These results show that the7