HEALTH AND NUTRITION NEWSNo. 10December 15, 2004

Table of Contents
CURRENT RESEARCH PROJECTS
Focusing On What We Call Caspian Sea Yogurt……………………………………………Tatsuya Ishida
THOUGHTS ON HEALTH AND NUTRITION RESEARCH
Can Intake Of Antioxidants Reduce Immune System Ageing?............................................ Naomi Aiba
RESEARCH FINDINGS
Skeletal muscle FOXO1 (FKHR)-transgenic mice have less skeletal muscle mass, down-regulated
type I (slow twitch / red muscle) fiber genes, and impaired glycemic control………… Yasutomi Kamei
Effective Health and Nutrition Materials towards Occupation Group…………………. Katsushi Yoshita
Gene Mest Which Enlarges Adipose Cell..........................................................................Osamu Ezaki
Women with high BMI tend to underreport energy intake –Study for new female university students
aged 18-20 years–………………………………………………………………………………Hitomi Okubo
Increasing Thinness among Young Women…………………………………………….. Hidemi Takimoto© National Institute of Health and Nutrition

Current Research Projects
Focusing On What We Call Caspian Sea Yogurt
Tatsuya Ishida
Division of Applied Food Research

In the Division of Applied Food Researchwhere I belong to, surveys examinations andtests of nutrient and other composition, regarding
registration and approval of Food for Special
Dietary Uses such as foods for the ill sick diet or
food products for specified health use, areconducted. And researches of effects influencesof nutrients and other food components of food toon the human body are on going in large part. I
am in charge of all the activities regardingorganisms such as measurement of vitamin B6,
vitamin B12, and niacin using bacteria, or
counting of bacteria such as bifidobacteria or
lactic acid bacteria in yogurt of food for specified
health use or lactic acid bacteria beverage. And
we also conduct research on what we call
Caspian Sea Yogurt as related activities.
Some of you may know that Caspian Sea
Yogurt is the yogurt that Dr. Yukio Iemori,professor of emeritus at Kyoto University broughtback from Caucasia. It has feature of lessacidity and sticky which differs from other yogurt
on the market. Also, it is accepted in each family
and people have a broad interest in handing onby repackaging into smaller because it is easy to
produce by transferring some portion to milk.
Then I looked up the feature of the bacteria.
First of all, I tried identification of the
including bacteria by testing DNA sequences andhomology of base sequence and DNA. Twokinds of bacteria, Lactococcus lactis subsp.
cremoris and Acetobacter orientalis were found.
Generally, bifidobacteria Bifidobacterium bifidum,
lactobacilli Lactobacillus, and Streptococcus
thermophilus are mainly included in yogurt andlactic acid bacteria beverage.
Bacteria of
Caspian Sea Yogurt differs differ from any of
those. Furthermore, past examples of reports
on source and ground of separation isolation
about these bacteria indicate that Lactococcus
lactis subsp. cremoris and Acetobacter orientalis
are were mainly separated isolated from starter ofcheeseandsoybean-fermented
food,respectively. Both were founded as bacteria
which were separated isolated from food.
However, Acetobacter orientalis have been
separated isolated only in Indonesia up till now,and the idea that there is no example ofseparation isolation from daily products generates
interests to our future researches.
And
Lactococcus lactis subsp. cremoris are
antibacterial had antimicrobial activity against to
some of other lactic acid bacteria.
In the first place, I got attracted to this yogurt
by accident. I have come to work for thisinstitute since two years ago, and conducted theresearch on probiotex probiotics including lactic
acid bacteria bacterium. In the meantime, I havemainly done the research on evolution andsystem phylogeny of cyanobacteria (blue-green
bacterium bacteria: the first organisms on earthwhichcarriedout
oxygengenerating
photosynthesis), and I also have studied oncommon bacteria germ, not taking particular noteof lactic acid bacterium bacteria. When I had a
hard time deciding subject of my study on that
account, prior co-workers introduced me CaspianSea Yogurt, saying “try some interesting yogurtthat I eat at home”. When I tried it, it was had
viscosity thick, no acidity, and aroma of yeast. I
haven’t seen any yogurt like it. But I just felt thiswas an interesting fermented daily milk product,
and kept adding milk on it so that I never thoughtof as reporting like this.
By the way, aroma of Caspian Sea Yogurtthat I’ve got from prior co-worker seemed a little
differ from one I am researching now since origin
were different so that other microorganisms germmay existed. Although I really want to find themout, I didn’t realize to lost it, I’m ashamed to say.
It’s really too bad that it is difficult to take a look,but I want to check it out when I get rare
fermented daily product some day.

2

Thoughts on Health and Nutrition Research
Can Intake Of Antioxidants Reduce Immune System Ageing?
Naomi Aiba
Division of Applied Nutrition

According to abridged life table for 2003,
reported in July this year, Japan is the country
where people have the longest life expectancy inthe world in which current life expectancy for men
and women are 78.6 and 85.33 years old
respectively.
When we look at the life
expectancy table all over the world, women live
longer than men in each country. Why is there
gender difference in life expectancy around theworld? Since causes of death are based on
various factors such as biological, sociological,
environmental and so forth, it seems impossible
to state just the single reason for thisphenomenon. But immune system is necessary
for people’s health maintenance, and lowering the
mortality rate by infectious diseases greatly
involves in prolonged life expectancy, so we aregoing to take a look at this gender difference oflife expectancy from viewpoint of immune system.
Immunity starts from distinguishing between
self and not-self, and it excretes not-self. In thiscase, for example, not-self is the source of
infection (virus or bacteria) or infected cell in
infectious disease, and cancer originated in the
body is regarded as same. If not-self can’t berecognized properly, the immune system doesn’t
work properly. As a result, various diseases aredeveloped, and cure will be difficult. Here is areason why the immune function is cited as a big
factor which influences on the fate of organisms.
The immune function is mainly found incell-mediated immunity, especially in the Tlymphocyte
functions.
Tlymphocytes
differentiate and proliferate within the thymus,
and move to peripheral blood and lymph node.
The thymus, in which the T lymphocytes
differentiate and maturate, gets bigger during
Table
countryJapan
Hong Kong
Switzerland
France
Italy
Iceland
CanadaSwedenNorwaymen

growthperiod.
Afterthematuration,
age-associated thymic atrophy occurs and resultsin the inefficient functioning of the immune system.
In this regard, there is a gender difference on aperiod when thymic atrophy occurs, and thymic
atrophy with male apparently occurs in earlier age,suggesting that the decline of immune function inmale occur earlier than that in female. Thymic
atrophy can be treated as one of the signs ofaging, and immune system ageing in male occurs
earlier than that in female. Intracellular reactive
oxygen species are thought as candidates forageing causes of cells including thymocyte.
Both elimination of reactive oxygen species andantioxidant capacity towards reactive oxygen
species are known to influence on aging andfalling lifestyle related diseases.
Both theactivities of antioxidant enzymes and antioxidant
components have antioxidant ability within an
organism, and both can be controlled by various
nutrition components in foods.
Antioxidant
enzymes need metals such as selenium, zinc,and magnesium for their activities and also theintake of antioxidant-related nutrients canenhance antioxidant state within the cell.
Therefore, antioxidant components in the foodare focused as an anti-ageing factor. Recently,
the state of intracellular antioxidant is reported toinfluence toimmune balance and antioxidant
components are expected to be a factors toregulate immune function. Proper intake of foodcontaining antioxidants may control thymic
atrophy and maintain immune system, and theearly decline of immune function with male may
be delayed. Eventually, it may realize that men
have the same level of life expectancy as women.

Life Expectancy by countries and genders
women

difference3

country
Australia
Germany
England
South Korea
USA
China
Russia
India

men

women

differenceResearch Findings
Skeletal muscle FOXO1 (FKHR)-transgenic mice have less skeletal muscle mass,
down-regulated type I (slow twitch / red muscle) fiber genes,
and impaired glycemic control.
Yasutomi Kamei
Division of Clinical Nutrition

Skeletal muscle is the largest organ in thehuman body, comprising about 40% of bodyweight. The mass and composition of skeletal
muscle is critical for its functions, such as
exercise, energy expenditure and glucose
metabolism. Skeletal muscle mass andcomposition are plastic and affected by physicalactivity, environmental factors, or pathologicalconditions; for example, skeletal muscle mass is
reduced by lack of muscle use in a long-term
bedridden condition. This skeletal muscle loss iscalled atrophy. Atrophy is also observed during
starvation, cachexia, severe diabetes and aging.
However, the precise mechanism of atrophy isunclear.
FOXO1, a member of the FOXO forkhead
type transcription factors and co-factor of nuclear
hormone receptor, is markedly up-regulated inskeletal muscle in energy-deprived states such as
fasting and severe diabetes, but its functions inskeletal muscle remain poorly understood. In thisstudy, we created transgenic mice specificallyoverexpressing FOXO1 in skeletal muscle. These
mice weighed less than the wild-type control mice,
had a reduced skeletal muscle mass, and themuscle was paler in color (Fig. 1). Microarray
analysis revealed that the expression of many
genes related to the structural proteins of type I
muscles (slow twitch, red muscle) weredecreased. Histological analyses showed amarked decrease in size of both type I and type II
fibers, and a significant decrease in the number ofControl

type I fibers in the skeletal muscle of FOXO1 mice.
Enhanced gene expression of a lysosomal
proteinase, cathepsin L, which is known to be
up-regulated during skeletal muscle atrophy,
suggested increased protein degradation in theskeletal muscle of FOXO1 mice. Running wheel
activity (spontaneous locomotive activity) was
significantly reduced in FOXO1 mice compared tocontrol mice. Moreover, the FOXO1 mice showed
impaired glycemic control after oral glucose andintraperitoneal insulin administration.
Then, we examined the change inendogenous FOXO1 expression caused byphysical inactivity. The right hindlimbs of wild typemice were immobilized in plaster casts, and theleft hindlimbs were left freely moving as a control.
After three weeks in the plaster casts, the skeletal
muscle weight of the right hindlimbs was
significantly lighter than that in the controls. The
expression of type I genes, but not type II genes,
was markedly decreased in the plaster-casted
muscle. At the same time, endogenous FOXO1mRNA was increased in the immobilized muscle.
Namely, the change in gene expression observedin the plaster-casted skeletal muscle was similar
to that observed in the FOXO1 mice. These
results suggest that FOXO1 negatively regulates
skeletal muscle mass and type I fiber gene
expression, and leads to impaired skeletal muscle
function. Activation of FOXO1 appears to be
involved in the pathogenesis of muscle atrophy.

FOXO1 mice

Tibial anterior muscle
Extensor digitorum longus

Turning red muscle

SoleusFigure2. A schematic model.
Figure1. Photo of “FOXO1 mice” muscle.

PGC-1α causes type I fiber formation. As the FOXO1
protein can interact with the PGC-1α protein, FOXO1
may affect certain functions of PGC-1 α . A certain
nuclear receptor(s) and transcription factor(s), which
can interact with both FOXO1 and PGC-1α, may be
involved in a process positively and negatively regulatedby PGC-1α and FOXO1.

Soleus and tibial anterior muscle of control mice are red(left), while those of FOXO1 mice are paler (right).
Extensor digitorum longus muscle is composed of type II
fibers, less change was observed between control andFOXO1 mice.

4

Research Findings
Effective Health and Nutrition Materials towards Occupation Group
Katsushi Yoshita
Division of Health and Nutrition Monitoring

Appropriate nutrition intake can not be lacked
to maintenance and promotion of good health,and prevention of disease. Thus, it is necessary
for subjects of individuals and groups to learn andpractice preferable diet life and lifestyle. And it isnecessary to convey the right information. But
kinds of materials to gain more attention andeffective are hardly exposed. So we provided 3
kinds of information of health and nutritionmaterials, and attract attentions.
Methods of information service are following
3 materials:
1) Point of purchase advertising menus (POPmenus)
A5 size, horizontal, colored, written on themenu holder.
Change contents every week, set on every
table of staff’s dining room.
Basic configuration of space is title or promo
word at upper 15% part, comments at 45% of
page, and related illustrations at rest of 40%.
2) Posters
A3 size, vertical, colored.
Change contents every 4-8 weeks.
Show at appointed places in the businessplace.
Place text at 40-50% of page, related
illustrations and figures at the rest.
3) Leaflets
A5 size, vertical.
Distribute at the event and campaign.
Use colored papers other than white papers
to attract people’s attention.
Ratio of letters, illustrations, figures can bechanged depends on topics
At any hand, contents are easily
understandable by most employees based on
preliminary study. After at least 1 year from thestart of information service, we conductedquestionnaire survey to occupation group, anddetermine its coverage.
Among 3 study materials, material which
attracted a lot of attention was POP menus.
While some differences occurred in each field,
about 50% of those polled answered that theyread them all or pretty much with update of thecontents. Especially women pay high attention
as over 60%. On the other hand, only 35% ofpoles answered that they read all or most of
posters with it’s’ update, and about 15%
responded that they didn’t read at all. And only
15% of men and 20% of women replied that theyread leaflets through every time the contents
renewed.
POP menus was recognized as highly
effective because many of them were set to comein sight for sure, and kept interests of dining room
users by changing its contents sometimes atvarious intervals. On the other hand, dramaticeffects of posters or leaflets were not always seen.
It seems there were negative impacts of limitingtime and space of posting and distributing, andinconvenience of moving around to see them.
The results of this time show necessity forthe select and use of adequate materials
according to the situation, not by just sending out
health and nutrition information unilaterally.

Tips on Eating Vegetables As Much As You Can!
••

Point of Purchase advertising menus (POP menus)
5

Vegetables get smaller in size by cooking so thatyou can eat great quantity easily.
While nutrients are lost by heating, it is actually
efficient if you consume a lot.

Research Findings
Gene Mest Which Enlarges Adipose Cell
Osamu Ezaki
Division of Clinical Nutrition

Obesity is said to be becoming a breeding
ground for Type 2 diabetes, arteriosclerosis,
hypertension, and hyperlipemia. Why is it so,then?
Obesity is a condition accumulatingexcessive body fat, not just overweight but
rather a condition increasing a number ofadipose cell, or a condition that adipose cell
itself gets bigger.
Type 2 diabetes,
arteriosclerosis, hypertension, and hyperlipemia
are especially developed when abdominal
visceral fat cell that liver and intestines exist
enlarges.
Adipose cell produces various
physiologically active substance or hormonelike
substance as endocrine organ.
These
substances are generally called adipocytokines,
and related to regulation of sugar metabolism or
fat metabolism in the body. But enlarged
adipose cell produces unusual amount of bad
adipocytokine, and cause various diseases by
disturbing
sugar
metabolism
andfat
metabolism.
In other words, a key to exploring the causeof obesity as a breeding ground for disease isadipose cell. Then we examined mice to knowwhat kinds of gene vary in visceral fat cell. We
found that gene called Mest was significantly
expressed in overweight mice by high-fat diet, or
genetically overweight and diabetic mice. And
we conducted various tests to know how Mest

works in adipose cell.
As a result, Mest didn’t express in tissue
other than adipose cell, and it only significantly
expressed in adipose cell of fat mice. Obesity
often accompanies abnormality in the metabolism
of sugar, and Mest is deeply associated withobesity itself rather than regulation of blood sugar.
On the other hand, tests on intake of pioglitazone,
drug for Type 2 diabetes proved that expressionof Mest reduced when enlarged adipose cell got
smaller. In addition, tests on cultured cell of overexpressed Mest promotes expression of specificgene including adipocytokine gene in adipose cell.
We created transgenic mice that overexpressed
Mest in adipose cell, and evaluated activity of
Mest in the body. Then this transgenic miceincreased expression of gene specific for fat, andenlarged adipose cell.
Consequently, Mest
came out not only in enlarging adipose cell but
also increasing expression of bad adipocytokine.
To put it the other way around, control expressionof Mest suggests preventing enlargement of
adipose cell and abnormal secretion of
adipocytokine. For these reasons, creating adrug targeting on Mest may be effective toprevent obesity or various diseases thatassociates with this.

adipose cell of original miceadipose cell of transgenic mice

Figure:Enlargement of adipose cell on Mest transgenic mice6

Research Findings
Women with high BMI tend to underreport energy intake– Study for new female university students aged 18-20 years –Hitomi Okubo
Project of Scientific Evaluation of Dietary Reference Intakes

We frequently hear contradictory commentsfrom weight-conscious people as to “although noeating, gaining weight”, or receives answers fromobese people at dietary survey like “no eating”
while they must’ve been eaten. These concernsa phenomenon called ‘underreporting’ that is
estimating less than amount of actual intake, andone of the problems that dietitians bother about.
However, reports on this problem are mostly
developed by the United States or Europe, andthere is very few study for Japanese. So weevaluated reporting accuracy of energy intake
among Japanese, and examined the feature of agroup which likely underreport.
This survey was conducted based on thecooperation of female university students aged
18-20 y who entered a college in 1997. Two kindsof questionnaires were prepared, and they wereasked about foods they have eaten within one
month and their lifestyle. We calculated the ratioof energy intake (EI) to basal metabolic rate(BMR), EI/BMR whether their reports of energy

Graph2.

Nutrient intake of each group, compared togroup Ⅴ in which EI/BMR is the largest (%)

Relationship between energy intake(EI/BMR) by self-assessment & BMI

2

Average BMI (kg/m )Graph1.

intake were reasonable. It indicated that the lower
the EI/BMR and the lower the reports of energy
intake.
In graph 1, the subjects were divided into fivegroups by this ratio, and compared their BMI. The
lower EI/BMR group had the higher BMI. In graph2, it shows the nutrient intake gradient according
to the level of underreporting. Compared to thegroup Ⅴ which reported the highest energy
intake, the lower energy intake groups tended toreport less intake of fat and protein, and more ofcarbohydrates. These results indicate that womenwhose BMI are high tend to underreport.
Reporting macro nutrient intake balance may also
be correlated with accuracy of reporting energy
intake.
Subjects of this study are female university
students aged 18-20 y, and the results may not be
always true for all Japanese. However, we needto keep it in our mind that ‘underreporting’ may
exist in obese people.

H

L*

Relationship between energy intake(EI/BMR) by self-assessment &nutrient intake

carbohydrates
protein

lipid

L

H

Ratio of EI and BMR

Ratio of EI and BMR

Difference Test with EI/BMR of groupⅤ;
One-way analysis of variance (Dunnett’s)

Difference Test with EI/BMR of groupⅤ;
One-way analysis of variance (Dunnett’s)

7Research Findings

Increasing Thinness among Young Women
Hidemi Takimoto
Center for Collaborative Researchthan 18.5 kg/㎡ as “underweight.”
Figure shows ratio of BMI and thinness by
age for 25 years.
As the figure shows,significant decline in average BMI, and significant
increase of thinness, can be seen in
15-29-year-old women. In particular, in a groupof 25-29-year-old women, ratio of thinness
increased significantly from 13.5% during
1976-1980 to 23.7% during 1996-2000. During
the survey period of 25 years, growth of averageheight of all ages increased significantly while
average weight didn’t change very much.
Why did thin women increase like this? As
one of the reasons of this, increase of eating
disorder such as anorexia can be assumed. But
frequency in Japan is estimated as 17~30 womenin 100,000 women, and it is low compared to
270women in the US and 115 women in the UK.
Another reason we can think of is increase insmoking rate. From the results of national
nutrition survey, smoking rate of 20-year-old
women was 11.9% in 1990, and it almost doubledof 20.9% in 2000.
From now on, we need to conduct a further
investigation into the reasons why thin women
increased like this, and investigate healthproblems (anemia, irregular menstruation and soforth) that thinness women are likely be affected.

Obesity is one of the major health problems
on a worldwide scale.
Not a single day
passes by without hearing the news about healthrisks due to obesity in the mass media. We fearthat too much strength on obesity prevention
might lead young women who have slimming
obsession to unnecessary weight loss.
Excessive weight loss is pointed out as causing
nutritional deficiency, irregular menstruation, andeating disorders. In developed countries, healthrisk of underweight is not as popular as that ofoverweight. But a cohort study on middle-aged
and senior adults showed that the mortality of“underweight” women was higher than one of“normal” weight women. And women who were
underweight before pregnancy have a higher riskof intrauterine growth retardation or prematurebirths. Then, is there an evidence of thinness inyoung women increasing?
We used data on height and weight in 30,903
non-pregnant and non-lactating women aged
15-29- year-old, who participated in the National
Nutrition Survey in 1976-2000, and examinedchange during the 25 years. Subjects weredivided into 3 groups as 15-19-year-old,
20-24-year-old, 25-29-year-old, and terms weredivided into 5 groups of 5 years each. We used
standards of body mass index from Japan Society
for the Study of Obesity, and judged BMI of less15-19 y thinness
20-24 y thinness
25-29 y thinness
15-19 y ave. BMI
20-24 y ave. BMI
25-29 y ave. BMI

8