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Alteration of intestinal flora by the intake of enzymatic degradation products of adlay (Coix lachryma-jobi L. var. ma-yuen Stapf) with improvement of skin condition


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Adlay has been used as a traditional Chinese medicine and nutrient for its beneficial effects on bowel movements and skin care. This study examined the effect of enzymatic degradation product of adlay, “Super Hatomugi” (SPH) on human skin and the intestinal flora in a randomized, double-blind placebo-controlled study. The subjects were divided into three groups: 500 mg SPH, 1000 mg SPH, and placebo, taken daily for 4 weeks. Hematological and skin condition examinations as well as an analysis of intestinal flora were performed 2 weeks before and 10 weeks after the start of the SPH intake. Skin condition was improved by SPH intake as revealed by a reduction in the number of nucleated epidermal cells. In addition, an increase in the fecal population of Bacteroidetes followed the SPH intake. These results show the possibility that SPH improves the skin condition and changes the proportions of intestinal flora.
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Alteration of intestinal flora by the intake
of enzymatic degradation products of adlay
(Coix lachryma-jobi L. var. ma-yuen Stapf) with
improvement of skin condition
Mie Nishimura
, Tatsuya Ohkawara
, Hiroyo Kagami-Katsuyama
, Shizuko Sekiguchi
Toshio Taira
, Mana Tsukada
, Hiroki Shibata
, Jun Nishihira
Department of Medical Management and Informatics, Hokkaido Information University, Ebetsu 069-8585, Hokkaido, Japan
Primary Cell Co., Ltd, Sapporo 001-0021, Japan
HABA Laboratories Inc., Tokyo 101-0041, Japan
Section of Research and Development, Hokkaido Food Industry Promotion Organization, Sapporo 060-0001, Japan
Article history:
Received 12 August 2013
Received in revised form
6 January 2014
Accepted 9 January 2014
Available online xxxx
Intestinal flora
Nucleated epidermal cell
Skin texture
Adlay has been used as a traditional Chinese medicine and nutrient for its beneficial effects
on bowel movements and skin care. This study examined the effect of enzymatic degrada-
tion product of adlay, ‘‘Super Hatomugi’’ (SPH) on human skin and the intestinal flora in a
randomized, double-blind placebo-controlled study. The subjects were divided into three
groups: 500 mg SPH, 1000 mg SPH, and placebo, taken daily for 4 weeks. Hematological
and skin condition examinations as well as an analysis of intestinal flora were performed
2 weeks before and 10 weeks after the start of the SPH intake. Skin condition was improved
by SPH intake as revealed by a reduction in the number of nucleated epidermal cells. In
addition, an increase in the fecal population of Bacteroidetes followed the SPH intake. These
results show the possibility that SPH improves the skin condition and changes the
proportions of intestinal flora.
2014 The Authors. Published by Elsevier Ltd. All rights reserved.
1. Introduction
Skin aging is a complex biological process influenced by
a combination of endogenous and exogenous factors
(Debacq-Chainiaux, Leduc, Verbeke, & Toussaint, 2012). A
variety of bioactive components derived from plants such
as vitamins, amino acids and polyphenols have been re-
ported to support the health and beauty of skin (Martorana
et al., 2013; Schagen, Zampeli, Makrantonaki, & Zouboulis,
2012), and thus foods and supplements derived from these
plants have been of particular interest in terms of health
1756-4646/$ - see front matter 2014 The Authors. Published by Elsevier Ltd. All rights reserved.
5This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-No Derivative
Works License, which permits non-commercial use, distribution, and reproduction in any medium, provided the original author and
source are credited.
*Corresponding author. Tel.: +81 11 385 4411; fax: +81 11 384 0134.
E-mail address: (J. Nishihira).
Available at
journal homepage:
Please cite this article in press as: Nishimura, M. et al., Alteration of intestinal flora by the intake of enzymatic degradation products of adlay (Coix lachryma-jobi L.
var. ma-yuen Stapf) with improvement of skin condition, Journal of Functional Foods (2014),
Adlay (Coix lachryma-jobi L. var. ma-yuen Stapf) is an annual
crop cultivated mainly in India, China and Japan (Hu, Zhao,
Liang, Qiu, & Chen, 2007). Adlay seeds have been used widely
as a traditional Chinese medicine for the treatment of edema,
rheumatism, and neuralgia.
Adlay was found to have various pharmacological effects.
For example, rats fed adlay oil showed significant decreases in
serum insulin and leptin levels (Huang, Chiang, Yao, &
Chiang, 2005). In rats with hyperlipidemia, adlay seed oil in-
take reduced the volume of abdominal fat tissue and the level
of low-density lipoprotein concentration, and increased the
total antioxidant capacity (Yu, Gao, Zeng, & Liu, 2011). These
reports suggest that adlay may help ameliorate metabolic
syndrome, a cluster of conditions that is characterized by
abdominal obesity and hyperlipidemia. In addition, other re-
search groups have revealed that dehulled adlay reduced
the risk of colorectal carcinogenesis through the modulation
of COX-2 expression in a rat model (Hung & Chang, 2003;
Shih, Chiang, & Kuo, 2004).
The consumption of adlay has also been thought to pre-
vent or improve various skin diseases such as dry skin, warts
and atopic dermatitis. Adlay’s therapeutic effects for skin dis-
orders are speculated to be based on the recovery of skin bar-
riers by an increase in ceramide (Huang, Chung, Kuo, Lin, &
Chiang, 2009). It is also speculated that adlay exerts anti-aller-
gic effects by producing a shift in the Th1/Th2 balance from
Th2 to Th1 dominance (Hsu, Lin, Lin, Kuo, & Chiang, 2003).
Several studies have indicated that the intestinal flora
play an important role in obesity and cancer, as well as
inflammatory bowel disease (IBD) (Bajzer & Seeley, 2006;
Bien, Palagani, & Bozko, 2013; Jobin, 2013). It is now empiri-
cally recognized that an individual’s skin condition can re-
flect the environment of his or her intestinal flora (Bisgaard
et al., 2011); however, the influence of the intestinal flora
on the skin’s condition has not been well investigated.
Concerning the change of intestinal flora in association with
adlay intake, rats fed adlay showed a significant change in
the population of lactic acid bacteria in faeces (Chiang,
Cheng, Chiang, & Chung, 2000). Experimental data regarding
the relationship between the proportion of intestinal flora
and the skin’s condition in humans could provide valuable
information about the skin’s condition in light of the pres-
ence of microbiota.
Recent studies have demonstrated that adlay contains a
variety of biologically active compounds including oligosac-
charides and polyphenols that may improve bowel functions
(Son, Kim, & Lee, 2008) and have an antioxidative effect
(Wang, Sun, Yi, Wang, & Ju, 2012), respectively. Unfortunately,
adlay extracts lose much of their bioactive compounds
through the process of extraction and purification. We thus
successfully developed ‘‘Super Hatomugi (SPH),’’ in which
whole adlay is crushed and degraded by enzymatic treatment
with a-amylase, neutral protease and transglucosidase. We
found that SPH contained much higher amounts of free ami-
no acids, polyphenols, and oligosaccharides compared to ad-
lay extracts prepared by conventional methods. To test the
efficacy of SPH for human health and skin conditions, we car-
ried out a clinical intervention focusing on the condition of
the skin in association with intestinal flora.
2. Methods
2.1. Study subjects
Sixty-eight healthy female volunteers (age 24–47 years) were
recruited. None had a history of recent gastrointestinal disor-
ders, pregnancy, significant disease, surgery, severe allergic
reaction to food, or current use of any medication. The sub-
jects’ body weights and body mass index (BMI) values are
listed in Tab le 1.
The clinical intervention was carried out as a randomized,
double-blind placebo-controlled trial. At randomization, the
68 eligible subjects were randomly and blindly assigned to
one of three treatment groups: 1000 mg SPH, 500 mg SPH, or
Placebo. For 4 weeks, each night before going to bed, the sub-
jects took 1000 mg SPH or 500 mg SPH or placebo (glucose)
contained in six capsules. Physical and hematological exam-
inations as well as VAS (Visual Analogue Scale) score mea-
surements were performed at 2 weeks before, baseline
(0 week), and 4 weeks after the supplementation started.
Measurements of the enteric bacteria profile and skin condi-
tion were carried out twice, at 2 weeks before and 4 weeks
after the supplementation started.
All subjects provided written informed consent prior to
undergoing any study-related tests, and the protocol was ap-
proved by the Ethics Committee of Hokkaido Information
University. The study protocol was conformed to the Helsinki
2.2. Super Hatomugi (SPH)
The SPH was prepared by the use of adlay (Coix lachryma-jobi
L. var. ma-yuen Stapf) harvested in Japan without agricultural
chemicals. Adlay seeds with shell were crushed and sterilized
at 141–156 C for 3 min, and then crushed to the particle size
of 455 lm or less. The ground product (400 kg) was suspended
in water (3600 L) and treated with 25% sodium hydroxide
(18 L) for 3 h at 75 C for gelatinization of starch. After pH
adjustment between 5.45 and 5.55, the suspension was trea-
ted with a-amylase, SumizymeL
(New Japan Chemical
Industry Co. Ltd., Tokyo, Japan) and neutral protease, Sumizy-
(New Japan Chemical Industry Co. Ltd., Tokyo, Japan)
at 50–52 C. After the enzyme treatment for 3 h, the
product was further treated with transglucosidase (Amano
Table 1 – Characteristics of the subjects in the placebo,
500 mg SPH and 1000 mg SPH groups.
Characteristic Placebo 500 mg SPH 1000 mg SPH
Number of subjects n=22 n=22 n=22
Age (years) 37.77 ± 6.29 37.91 ± 5.46 38.68 ± 5.27
Body weight (kg) 52.96 ± 7.60 51.65 ± 6.23 51.21 ± 7.70
Height (cm) 158.32 ± 6.71 158.76 ± 6.63 157.81 ± 4.30
Body mass index
21.10 ± 2.39 20.54 ± 2.23 20.46 ± 3.45
Values shown are mean ± SD. Statistical analysis was performed by
one-way analysis of variance.
Please cite this article in press as: Nishimura, M. et al., Alteration of intestinal flora by the intake of enzymatic degradation products of adlay (Coix lachryma-jobi L.
var. ma-yuen Stapf) with improvement of skin condition, Journal of Functional Foods (2014),
Pharmaceutical Co., Ltd., Aichi, Japan) to obtain isomaltooli-
gosaccharide. After enzyme inactivation, the soluble product
was concentrated to Brix degrees of 19–22. The final product
was prepared by drying and subsequent powdering and
encapsulation. Analytical results of the composition of SPH
compared to adlay, and coix seed are provided in Tab l e 2.
The production and the packing were carried out in HABA
Corporation (Tokyo, Japan), under the quality-controlled man-
ufacturing plant in compliance with the Food Sanitation Act
(the Ministry of Health, Labor, and Welfare of Japan). The
quality and safety of test samples were thoroughly examined
by HABA Corporation.
Supplementation of SPH for healthy human volunteers
was carried out at doses of 1000 mg/day for 4 weeks. There
were no side effects caused by supplementation of SPH. Based
on these results, it is considered that SPH is tolerant of hu-
mans at the doses of 1000 mg/day for 4 weeks.
2.3. Preparation of corneocytes
Skin of 21 randomly selected subjects was tape-stripped for
the measurement of nucleated epidermal cells. Cellophane
film (Promotool Corp., Tokyo, Japan) was firmly attached to
the skin on the subject’s cheek and gently pressed by hand
over the entire area. The removed films were adhered to slide
glasses, and these samples were analyzed using an inverted
microscope, IX71 (Olympus, Tokyo, Japan). Nucleated epider-
mal cell numbers were calculated by viewing five visual fields,
and their averages were calculated.
2.4. Measure of sebum capacity, transepidermal water
loss and moisture
The sebum secretion of the face was measured using the
SM815 (C-K Electronics, Cologne, Germany), as
described (Sator, Schmidt, & Ho
¨nigsmann, 2003). In brief,
the amount of sebum secretion was recorded at the forehead.
To collect sebum, a plastic strip was applied at the site with a
constant pressure of 4 N for 30 s. The originally translucent
plastic strip becomes transparent when it absorbs sebum.
The Sebumeter
calculates the sebum level by measuring
the transparency of the strip using a photodetector. All mea-
surements were performed by the same investigator in a
room with a constant temperature and constant humidity.
The transepidermal water loss of the upper arm surface
and the moisture of the face was measured using the TEW-
CM825 (C–K Elec-
tronics, Cologne, Germany) according to the manufacturer’s
2.5. Real-time PCR for analysis of enteric bacteria
DNA from subjects’ fecal samples was extracted using a kit
(DNA Extraction kit, Qiagen, Valencia, CA, USA) according to
the manufacturer’s protocols. Phyla-specific primers were
used to quantify 16S rRNA gene copy numbers (rDNA) of Phyla
Bacteroidetes, Phyla Firmicutes and total bacteria. Real-time
polymerase chain reaction (PCR) was performed in a 20.0-lL
reaction containing 5.0 lL of DNA (<100 ng), 10.0 lLof2·Light
480 SYBR Green I Master (Roche Diagnostics, Mann-
heim, Germany), 3.0 lLofH
O, and 1.0 lL of 500 nM primers.
Amplification was performed on a Light Cycler
480 qPCR sys-
tem (Roche Diagnostics, Mannheim, Germany). The total bac-
teria primers were 50-ACTCCTACGGGAGGCAGCAGT-3 0and
50-GTATTACCGCGGCTGCTGGCAC-30for forward and reverse
primers, respectively. The Bacteroidetes primers were forward
primer 50-AGCTGACGACAACCATGCAG-30. The Firmicutes
primers were forward primer 50-GGAGYATGTGGTTTAATTC-
GAAGCA-30and reverse primer 50-AGCTGACGACAACCATG-
CAG-30. The percentage of Bacteroidetes or Firmicutes was
calculated as the ratio to the cycle number of total bacteria.
2.6. Analysis of serum contents of biomarkers
The subjects fasted overnight, and blood samples were drawn
before breakfast on days 0 and 28 for liver function and renal
function. The samples were separated immediately by centri-
fugation (1000·g, 10 min) and were stored at 80 C within
2 weeks until use. Clinical diagnostics tests for alanine ami-
notransferase, blood urea nitrogen and other biomarkers
were carried out using a Roche/Hitachi 912 chemistry ana-
lyzer (Roche Diagnostics, Mannheim, Germany).
2.7. Statistical analysis
Averages and standard deviations of age and other parame-
ters were calculated for each group. Statistical analyses were
performed with the program IBM SPSS Statistic 19 (IBM, Ar-
Table 2 – Component of Super Hatomugi (SPH) compared
with Coix seed, adlay per 100 g.
Component SPH Coix seed Adlay
Calories (kcal) 300 360 360
Carbohydrate (g) 66.9 72.2 72.2
Protein (g) 10.6 13.3 13.3
Fat (g) 5.5 1.3 1.3
Polyphenol (mg) 441 66 43
Dietary fiber (g) 28.9 1.7 30.6
Total free amino acid (mg) 364 35 24
Arginine 48 6 5
Lysine 14 2 2
Histidine 12 1 2
Phenylalanine 30 –
Tyrosine 26 –
Leucine 54 –
Isoleucine 14 –
Methionine 8 –
Valine 21 2
Alanine 32 5 4
Glycine 6 2 1
Proline 13 4 2
Glutamic acid 34 9 5
Serine 16 1 1
Threonine 11 –
Aspartic acid 22 3 2
Tryptophan 3 –
Cystine – –
Please cite this article in press as: Nishimura, M. et al., Alteration of intestinal flora by the intake of enzymatic degradation products of adlay (Coix lachryma-jobi L.
var. ma-yuen Stapf) with improvement of skin condition, Journal of Functional Foods (2014),
monk, NY, USA). The Wilcoxon signed-rank test was per-
formed for the data 2 weeks before and 4 weeks after the start
of the intake of adlay in each group. The statistical analysis of
the value changes of enteric bacteria was carried out by anal-
ysis of variance (ANOVA) with the post hoc Games–Howell
3. Results
3.1. Effect of SPH on skin texture
Sixty-six subjects completed the study; two subjects chose to
discontinue due to personal reasons before the supplementa-
tion began. There were no significant differences in age, body
weight, height, or BMI among the three subject groups (Ta-
ble 1). First, to determine the effect of SPH on the subjects’
skin, we used tape-stripping to measure the number of nucle-
ated epidermal cells and observed the subjects’ skin texture.
An increase in total nucleated epidermal cells indicates
abnormal cell turnover as a results of skin damage. At
4 weeks after the start of the supplementation, we observed
that the subjects who ingested SPH at the dose of 1000 mg/
day for 4 weeks had significantly decreased numbers of
nucleated epidermal cells (p= 0.018) (Fig. 1A). In addition,
improvement of the structure of skin texture following SPH
intake was demonstrated by microscopic analysis (Fig. 1B).
3.2. Effect of adlay on sebum secretion
We also examined the effect of SPH on sebum secretion and
transepidermal water loss and moisture. Although no signifi-
cant differences in the rates of skin moisture or transepider-
mal water loss were observed (data not shown), sebum
secretion was increased among the 1000 mg/day SPH sub-
jects, although not significantly so (p= 0.064) (Fig. 2).
3.3. Effect of SPH on intestinal flora
We analyzed the subjects’skin condition by evaluating the in-
crease of nucleated epidermal cells, an indicator of deteriora-
tion of the skin. In parallel, we examined the changes of the
subjects’ intestinal flora to evaluate the relationship between
skin condition and intestinal flora. We analyzed mainly the
fecal population of Bacteroidetes and Firmicutes. The popula-
tion of Bacteroidetes was slightly increased by SPH intake,
whereas little change of Firmicutes was observed (Fig. 3).
Consistent with this result, we found significant increases
in Bacteroidetes population for all 66 subjects (Fig. 4).
3.4. Levels of biomarkers of liver and renal functions after
SPH intake
We examined the levels of several biomarkers of liver func-
tion and renal function. Parameters for liver function, i.e.,
AST, ALT, and c-GTP, and those for renal function, i.e., BUN
and creatinine showed minimal changes after the SPH intake,
suggesting that SPH intake has no or minimal unfavorable ef-
fects on the liver and kidney, even at the dose of 1000 mg/day.
4. Discussion
Many disorders are associated with changes in the composi-
tion and metabolism of enteric flora (Guarner, 2006). Func-
tional foods, supplements and vegetables have been well
recognized as rich sources of the dietary fiber that exhibit
beneficial effects on health promotion through the improve-
ment of the balance of intestinal bacterial flora. It was re-
cently noted that dyspepsia might lead to skin disorders
(Roberfroid et al., 2010). In general, however, the correlation
between skin condition and intestinal flora is not well
Fig. 1 – Skin condition before and after Super Hatomugi (SPH) intake. (A) Quantitative measurements of nucleated epidermal
cells were carried out using tape stripping of subjects who completed a 4-week course of the daily intake of placebo (n= 8),
500 mg SPH (n= 6), or 1000 mg SPH (n= 7). Averages of 5 fields per sample were used. Data for 95th percentiles and medians
are shown.
p< 0.05 (2 weeks vs. 4 weeks) by Wilcoxon signed-rank test. (B) The images of corneocytes were captured by an
inverted microscope (400·).
Please cite this article in press as: Nishimura, M. et al., Alteration of intestinal flora by the intake of enzymatic degradation products of adlay (Coix lachryma-jobi L.
var. ma-yuen Stapf) with improvement of skin condition, Journal of Functional Foods (2014),
The results of the present double-blind and placebo-con-
trolled study demonstrated the potential effects of SPH on
the skin condition and the intestinal flora. We observed that
SPH improved the subject’s skin conditions, possibly through
a reduction of nucleated epidermal cells and improvement of
skin texture. In particular, an increase of the fecal population
of Bacteroidetes was clearly associated with the SPH intake.
Taken together, these results indicate that SPH could be a
beneficial supplement for the improvement of an individual’s
skin condition, possibly based on a change in the population
of intestinal flora.
Adlay has been reported to exhibit an anti-allergic func-
tion (Hsu et al., 2003) as well as an antioxidant action (Kuo,
Shih, Kuo, & Chiang, 2001), and therefore has been used in
China for the treatment of skin diseases (Chen et al., 2011).
Oxidative damage by reactive oxygen species (ROS) plays a
major role in skin aging (Oresajo, Pillai, Manco, Yatskayer, &
McDaniel, 2012; Ristow & Schmeisser, 2011). Antioxidants
such as carotenoids, tocopherols, and flavonoids, as well as
vitamins, have frequently been referred to as agents capable
of promoting skin health and beauty (Schagen et al., 2012).
Polyphenols with antioxidative activities were purified and
identified from adlay, including three lignan compounds
(syringaresinol, 4-ketopinoresinol, and mayuenolide) (Kuo
et al., 2002). Our previous study demonstrated that the meth-
anolic and dimethylsulfoxide (DMSO) extraction from SPH
had an antioxidant effect, and DMSO extraction reduced the
production of tumor necrosis factor-alpha (TNF-a; unpubl.
data). As shown in Fig. 1, we observed a reduction in the num-
ber of nucleated epidermal cells and an improvement of skin
texture following a 4-week SPH intake. We herein confirmed
that SPH have more than 10 times amounts of polyphenol
compared to adlay (Table 2 ). Although we did not investigate
kinds of polyphenol in SPH, these collective data suggest the
possibility that antioxidant action by polyphenols in SPH con-
tributes to the improvement of skin condition.
In addition, we developed SPH to enhance the amount of
free amino acid by protease treatment (Table 2). Free amino
acids are known to be critical components to collagen product
which are helpful for water retention properties of skin
(Rawlings & Matts, 2005). Furthermore, it has been demon-
strated that free amino acid are important not only as
Fig. 3 – Fecal populations of Bacteroidetes (A–C) and Firmicutes (D–F) compared with total fecal bacteria in the same subjects
who underwent tape-stripping for the measurement of the number of nucleated epidermal cells except for a few subjects
who showed abnormal bacteria values (i.e., the total ratio of Bacteroidetes and Firmicutes exceeded 100%). The data are
presented as data points for each subject before (2 weeks) and after (4 weeks) the start of the intake. Placebo group: (A)
(n= 7), (D) (n= 4); 500 mg SPH group: (B) (n= 5), (E) (n= 6); 1000 mg SPH group: (C) (n= 6), (F) (n= 6).
Fig. 2 – The effect of Super Hatomugi (SPH) on sebum
secretion after 4 weeks of the ingestion of placebo (n= 22),
500 mg SPH (n= 22), or 1000 mg SPH (n= 22). Data for the
95th percentiles and medians are shown. Statistical
analysis was performed by Wilcoxon signed-rank test
(2 weeks vs. 4 weeks).
Please cite this article in press as: Nishimura, M. et al., Alteration of intestinal flora by the intake of enzymatic degradation products of adlay (Coix lachryma-jobi L.
var. ma-yuen Stapf) with improvement of skin condition, Journal of Functional Foods (2014),
substrates for various metabolic pathways but also they have
a positive effect on signaling pathways (Meijer & Dubbelhuis,
2004). For example, L-threonine regulates meta-stability of
embryonic stem cells via PI3K/Akt, MAPKs, and mTOR
pathways (Ryu & Han, 2011). These results suggested that free
amino acid in SPH relate to the improvement of skin
condition as seen in polyphenols.
Inappropriate amounts of sebum from the glands can lead
to several skin disorders, such as acne and seborrhea (Borlu
et al., 2012). However, sebum also possesses the beneficial ef-
fects of preventing water loss from the skin surface and pro-
tecting the skin from infection by bacteria and fungi (Smith &
Thiboutot, 2008). In the present study, we found that the se-
bum capacity was increased by SPH intake. SPH intake also
significantly improved skin texture. We suspect that the SPH
intake improved the subjects’ skin condition by regulating
their sebum secretion.
Regarding the change in the population of microbiota due
to SPH intake, we observed that Bacteroidetes were increased
in the 1000 mg/day SPH group, in whom significant improve-
ment of the skin was also observed. Bacteroidetes are known as
the major population of the microbiota of animals, especially
in the gastrointestinal tract (Ishikawa et al., 2013). Members of
the Bacteroidetes have been known to act as opportunistic
pathogens, mostly causing post-operative infections and bac-
teremia (Murphy, Mo
¨rgelin, Cooney, & Frick, 2011). Moreover,
Bacteroidetes are reported to have the most effective antibiotic
resistance of any anaerobic pathogen (Kislak, 1972).
The mechanism underlying the increase in Bacteroidetes
may be due to the influence of oligosaccharides present in
SPH. Dietary oligosaccharides modulate the composition
and activity of intestinal microbiota. The intestinal fermenta-
tion of oligosaccharides leads to changes in the proportion of
intestinal microbiota and the increasing production of short-
chain fatty acids (Macfarlane, Steed, & Macfarlane, 2008;
Meyer & Stasse-Wolthuis, 2009). Bacteroidetes grew, induced
a-L-fucosidase activity, and produced abundant lactate or
short-chain fatty acid when fed oligosaccharides in vitro (Yu
et al., 2013). However, most oligosaccharides are converted
to monosaccharides and then absorbed from the small intes-
tine. We developed SPH to enhance the amount of oligosac-
charides, such as isomaltose and panose, dextrantriose,
from 8 to 15 g compared to adlay through a process in which
the carbohydrate component of adlay is converted to oligo-
saccharides by a-amylase and transglucosidase and degrada-
tion. Oligosaccharides avoid degradation and absorption in
the small intestine, which might lead to an increase in the
amount of Bacteroidetes in the colon. Moreover, in a prelimin-
ary study, we found that SPH enhanced the amount of dietary
fibers (28.9/100 g) more than coix seed (1.7/100 g) by the treat-
ment of transglucosidase. It is conceivable that SPH could
change the composition of intestinal microbiota through in-
crease of dietary fibers. This needs further investigation.
It was reported that supplementation with Bifidobacterium
breve, one of the microbiota, prevented UV-induced skin
damage through the suppression of the increases in both
elastase activity and interleukin (IL)-1blevels in the skin in
mouse (Sugimoto et al., 2012). This report supports the idea
that enteric flora alterations due to SPH intake could improve
the skin’s condition.
Consistent with these findings, we speculate that the
improvement of skin condition in the present SPH groups
may be mediated by the alteration of enteric flora in the colon
by oligosaccharides as well as other valuable effect by poly-
phenols, free amino acid. It is of interest to investigate the
mechanism of how SPH affects the population of intestinal
flora in association with the improvement of skin conditions.
The scores for abdominal symptoms were also assessed
with a Visual Analogue Scale (VAS); however, the subjects’
VAS scores were not significantly changed after SPH intake
(data not shown). We suspect that the administration period
was too short for the effect of fecal properties to be observed.
Fig. 4 – Changes in the ratios of fecal Bacteroidetes (A) and Firmicutes (B) compared with total fecal enteric bacteria after the 4-
week supplementation in all subjects except for the few who showed abnormal values (i.e., the total ratio of Bacteroidetes and
Firmicutes exceeded 100%, or the cycle threshold values (Ct) of >25.0). Values are means ± SE. The Games–Howell test was
p< 0.05. (A) Placebo (n= 12), 500 mg SPH (n= 12), 1000 mg SPH (n= 13). (B) Placebo (n= 8), 500 mg SPH (n= 12), 1000 mg
SPH (n= 14).
Please cite this article in press as: Nishimura, M. et al., Alteration of intestinal flora by the intake of enzymatic degradation products of adlay (Coix lachryma-jobi L.
var. ma-yuen Stapf) with improvement of skin condition, Journal of Functional Foods (2014),
We are planning to further investigate the effects of long-term
intake of SPH on quality of life (QOL).
In summary, the present findings demonstrated the poten-
tial efficacy of SPH for the improvement of skin conditions in
concert with changes in the microbiota population. However,
the underlying molecular mechanisms, including the identifi-
cation of the functional molecules in SPH, remain to be eluci-
dated. Nonetheless, the results of this clinical trial of SPH
broaden our knowledge of the functionality of adlay, particu-
larly SPH, and other functional foods for the improvement of
skin conditions and the promotion of health.
We thank Ms. Aiko Tanaka, Rina Kawamura, Tomoko Mino,
and Megumi Shibata for their technical assistance with the
data management, and Mr. Jungo Hayashi for his manage-
ment of the clinical trial. This study was supported in part
by the Organization of Hokkaido Food Innovation and the
Noastec Foundation.
Bajzer, M., & Seeley, R. J. (2006). Physiology: Obesity and gut flora.
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Please cite this article in press as: Nishimura, M. et al., Alteration of intestinal flora by the intake of enzymatic degradation products of adlay (Coix lachryma-jobi L.
var. ma-yuen Stapf) with improvement of skin condition, Journal of Functional Foods (2014),
... [48] Super Hamatogi (SPH) produced from coix by enzymatic reactions showed to have a positive effect on the development of human skin in a clinical trial . [95] Chapped skin, warts, eczema, and rheumatism were treated traditionally in China by coix . [1,38,47] The mechanism of the coix hydrolysis product to improve skin condition is by decreasing the number of nucleated epidermal cells, while severe acute radiation dermatitis occurred less frequently without side effects by the application of ethanol extract of coix . ...
... [1,38,47] The mechanism of the coix hydrolysis product to improve skin condition is by decreasing the number of nucleated epidermal cells, while severe acute radiation dermatitis occurred less frequently without side effects by the application of ethanol extract of coix . [7,95] The coix supplement can reverse and prevent the osteoporotic conditions in ovariectomized (OVA) mouse models applied in a certain dose . [66] The application of the coix diet and extract can increase calcium contents, alkaline phosphatase activity, and the density of bone minerals . ...
... [3] Qiao et al . [95] conducted an experiment for the development of inbred and hybrid using two coix varieties Coix lacryma-jobi L. and Coix lacryma-jobi L. friumentacea Makino). They observed heterosis on growth, height, leaf area, photosynthetic traits in the F1 generation, while pollen sterility, high pollen abortion rate (up to 79%), low seed setting rate (about 28%), and unstable hybrids were observed. ...
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Coix is a minor cereal crop with various applications in Traditional Chinese Medicine. Coix seed is called "Green Food", because of its high unsaturated fatty acid content with minimal toxic residues and heavy metals. Coix can be an excellent choice as a dietary supplement because of diverse bioactive compounds including phenols, flavonoids, polysaccharides, fibers, lipids, proteins, vitamins, and minerals with significant therapeutic effects such as anticancer, antioxidant, anti-inflammatory, anti-allergic, and immunomodu-latory, among others. The least attention has been given to the improvement of new elite cultivars of coix with high nutritional values and nutraceutical effects. To date, very few genomic studies, in particular, draft genome and de novo sequencing have been conducted to discover the genes responsible for nutritional traits and their molecular mechanisms that are insufficient for the efficient use of existing germplasms for molecular breeding of coix. Therefore, this review article attempts to highlight bioactive compounds identified from coix with their medicinal uses as well as the recent progress in coix genomic studies which will create research scope for the development of nutrient-rich coix varieties with high therapeutic values.
Skin inflammation and dryness are the features of surfactant-induced irritant contact dermatitis, a common skin disorder. In Japan, Coix seed (CS, Coix lacryma-jobi L. var. ma-yuen Stapf) is widely used as a traditional medicine and functional supplement to treat skin inflammation and dry skin. However, the efficacy of CS against surfactant-induced skin disorders has not been reported. Here, we investigated the effect of CS on inflammatory dry skin disorders induced by multiple topical applications of sodium dodecyl sulfate (SDS), a representative anionic surfactant. Male HR-1 hairless mice received a water extract of CS for four weeks. Three weeks after CS administration, the dorsal skin of the mice was exposed once daily to 10% SDS for five days. CS efficacy was then evaluated by measuring epidermal water content; erythema index; severity of skin scaling; epidermal thickness; inflammatory cell infiltration; production of pro-inflammatory mediators, such as interleukin-1α (IL-1α) and prostaglandin E2 (PGE2); and protein expression of cyclooxygenase 2 (COX-2), in the dorsal skin. Administration of CS markedly attenuated the SDS-induced reduction in epidermal water content, elevated erythema index, and severity of skin scaling. Histological analysis demonstrated that CS suppressed epidermal hyperplasia and macrophage infiltration in SDS-exposed skin. Furthermore, CS significantly prevented SDS-induced production of IL-1α and PGE2, as well as COX-2 upregulation. These results indicate that CS prevents SDS-induced inflammation-mediated skin dryness by inhibiting the production of pro-inflammatory mediators.
Coix seed (also known as adlay, Job’s tears) is a cereal grain mainly popular in tropical Asia. It is used in Chinese traditional medicine to treat various types of diseases and also consumed as food. It is rich nutritionally and contains a wide range of bioactive compounds and phytochemicals. The functionality of coix seed has been explored to the extent that it may provide a basis for its use as a food ingredient. Several studies have demonstrated the biological activity of coix seed. Published reviews on coix seed are scattered and not systematic. This review gives a comprehensive summary of the physicochemical composition, bioactivity, processing, application, and safety aspects of coix seed.
The aim of this study is to explore the hepatoprotective potential of coix seed protein hydrolysates (CPP) against alcohol-induced liver injury, and investigate the underlying mechanisms. The hepatoprotective activity of CPP at 0, 10, 30, 50 mg per kg BW was demonstrated in vivo by using ICR male mice fed with 40% v/v alcohol (5 ml per kg body weight) daily to induce alcoholic liver injury. CPP could significantly improve the alcohol metabolism in liver as evidenced by the enhanced activity of aspartate aminotransferase (AST) and alanine aminotransferase (ALT). The overexpression of serum tumor necrosis factor-α (TNF-α) and interleukin-β (IL-β) by alcohol induced injury was altered by CPP administration. The lipid peroxidation was also retarded by CPP by suppressing malondialdehyde (MDA) level and increasing the activity of liver superoxide dismutase (SOD). The findings from the present study suggested that CPP produced significant hepatoprotection and showed potential to be used as a dietary supplement or the ingredient of functional food.
The increasing need for food and better nutrition necessitates the investigation of different food sources. Among these, cereals are a major sector which provides most of the carbohydrates for the population. Cereals are classified as true cereals and pseudo-cereals based on their usage and classification. The true cereals have been given more attention and have been researched and used as food for a long time. The increase in the need for better nutrition necessitates discovering underutilized cereal sources, especially pseudo-cereals.
Barley (Hordeum vulgare L.) is the fourth most valuable cereal crop in the world exceeded only by rice, wheat, and corn [1]. Specifically, barley is the most widely adapted cereal grain species with production in a variety of extreme eco-agricultural areas, including regions with high latitudes, dry temperature, or severe temperature fluctuations such as Himalayan nations, Ethiopia, Tibet, and Morocco [2]. Yet 98% of barley crops is primarily used as animal feed and malting, while only 2% is used for direct food consumption.
Cyclodextrin (CD) inclusions are generally used to increase the solubility of poorly soluble drugs. In this study, magnolol (MAG) was used as a model drug for exploring the effects of CD on the degradation of pharmaceutical drugs by intestinal microflora. MAG/β-cyclodextrin (β-CD) and MAG/hydroxypropyl-β-CD (HP-β-CD) inclusion complexes were successfully prepared by the saturated aqueous solution and freeze-drying methods, respectively. Structural characterisation along with analyses of solubility, residual water content and drug content of the inclusion complexes was performed. The intestinal microflora of male rats was used to study MAG degradation in vitro. At three concentrations, the degradation of both the inclusion complexes was slower than that of the MAG monomer, MAG and CD mixtures and the MAG-poloxamer 188 micelle. There were no statistically significant differences in the degradation of the MAG/β-CD and MAG/HP-β-CD inclusion complexes. A simulation first-order equation of the degradation parameters revealed that the degradation of the inclusion complexes was slower and pronounced, judging by slope. The experimental findings were verified by molecular docking for predicting the stable molecular structure of the inclusion complexes. In conclusion, the inclusion complexes partially protected MAG from degradation by the intestinal bacteria.
A novel chymotrypsin inhibitor, named ClCI, was purified from coix seed (Coix lacryma-jobi L.) by aqueous two-phase extraction, chymotrypsin-Sepharose 4B affinity chromatography and centrifugal ultrafiltration. ClCI was a 7.9 kDa competitive inhibitor with pI 6.54. The inhibition constants (Ki) for bovine pancreatic chymotrypsin and bacterial subtilisin were 1.27 × 10-10 M and 1.57 × 10-9 M respectively. ClCI had no inhibitory activity against bovine trypsin and porcine elastase. ClCI had wide pH stability and good heat resistance. It can maintain >90% inhibition activity against chymotrypsin at 20-80 °C for 1 h. The primary structure of ClCI was highly similar (57%-92%) to those of several inhibitors belonging to the Gramineae crop potato protease inhibitor- I superfamily and showed the typical sequence motif of the protease inhibitor of the seed storage protein group. ClCI (12.5 mg) inhibited mycelial growth of the phytopathogenic fungi Mycosphaerella melonis, Helminthosporium turcicum, Alternaria solani, Phytophthora capsici, Isariopsis griseola, and Colletotrichum gloeosporioides, and caused 89% inhibition of the proteases from spore germination of plant-pathogenic fungi. The results of the present study indicate that ClCI had biotechnological potential as an alternative agent to combat the important phytopathogenic fungi.
The effects of six different species of edible mushroom (Auricularia auricular, Flammulina velutipes, Lentinus edodes, Agaricus bispours, Pleurotus osteratus and Pleurotus eryngii) on the intestinal microbial community composition and diversity in the fermentation liquor were investigated. During in vitro digestion and anaerobic culture, the short chain fatty acid (SCFA) concentrations and pH values were significantly altered in edible mushroom treated compared to those of control (P < 0.05). Moreover, the sequencing result showed that edible mushrooms had a great impact on the composition and diversity of intestinal microorganisms in fermentation liquor, which were structurally different compared with the control group (P < 0.05). Especially, A. auricular increased the abundance of Bifidobacteriales and Bacteroidales, decreased the abundance of Fusobacteriales; A. bispours increased the abundance of Lactobacillus. The effects among six kinds of mushrooms on intestinal microorganisms were significantly different in our exploratory study. In general, this study indicates that edible mushrooms have a positive role in gastrointestinal tract health by producing SCFAs and regulating the intestinal microbe.
Deacidification is one of the key steps in oil-refining processes. This study reported a simple approach for upgrading high-acid adlay bran oil by supercritical carbon dioxide (SC-CO2) extraction. The high-acid adlay bran oil was obtained by three-stage countercurrent ethanol extraction of adlay bran, with high free fatty acids (34 % FFAs). The FFA content in adlay bran oil reduced to 10.6 % by SC-CO2 extraction under optimized conditions. Interestingly, the SC-CO2 deacidification process also markedly reduced oil color value from 11.5 (Red) to 0.3 (Red) and acetone-insoluble matters in the oil from 1.56 % to 0.17 %. The results indicated solvent extraction combined with SC-CO2 deacidification method could be a feasible approach to significantly upgrade high-acid adlay bran oil and simplify refining process of adlay bran oil.
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Human milk oligosaccharides (HMOS) are not digested in the proximal intestine. In distal intestine, HMOS collectively modify the microbiota, but the response of individual bacteria to individual components of the HMOS is not well defined. Here, each of 25 major isolates of the human intestinal microbiota was fed individual major fucosylated and sialylated HMOS in anaerobic culture. This allowed for an assessment of the influence of specific HMOS on the growth and metabolic products of individual microbiota bacteria. Most Bifidobacteria spp. and Bacteroides spp. grew, induced α-l-fucosidase activity, and produced abundant lactate or short-chain fatty acids (SCFAs) when fed 2'-fucosyllactose (2'-FL), 3-FL, and lactodifucotetraose (LDFT). Lactobacillus delbrueckii ATCC7830, Enterococcus faecalis ATCC19433, and Streptococcus thermophilus ATCC19258 exhibited slight growth, pH reduction, and lactate production when supplemented with 2'-FL or 3-FL, but not LDFT. Supplementation with 3'-sialyllactose (3'-SL) and 6'-SL promoted moderate growth of Bifidobacterium longum JCM7007, 7009, 7010, 7011, 1272, 11347, ATCC15708, Bacteroides vulgatus ATCC8482, and B. thetaiotaomicron ATCC29148; accordingly, these bacteria exhibited greater neuraminidase activity and produced copious lactate, SCFA, or both. Lactobacillus delbrueckii ATCC7830 also consumed 6'-SL. In contrast, Clostridium spp., L. rhamnosus ATCC53103, E. faecalis ATCC29200, Staphylococcus spp., Enterobacter spp., and Escherichia coli K12 did not consume milk oligosaccharides nor produce appreciable acidic fermentation products. Specific Bifidobacteria and Bacteroides differentially digest specific individual HMOS, with the major fucosylated milk oligosaccharides most strongly stimulating key species of mutualist symbionts. This suggests strategies for treating dysbiosis of the microbiota and associated inflammatory disorders.
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Skin is a model of choice in studies on aging. Indeed, skin aging can be modulated by internal and external factors, reflecting its complexity. Two types of skin aging have been identified: intrinsic, mainly genetically determined and extrinsic-also called "photo-aging"-resulting on the impact of environmental stress and more precisely of UV rays. Simplified in vitro models, based on cellular senescence, have been developed to study the relationship between UV and aging. These models vary on the cell type (fibroblasts or keratinocytes, normal or immortalized) and the type of UV used (UVA or UVB).
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Skin has been reported to reflect the general inner-health status and aging. Nutrition and its reflection on skin has always been an interesting topic for scientists and physicians throughout the centuries worldwide. Vitamins, carotenoids, tocopherols, flavonoids and a variety of plant extracts have been reported to possess potent anti-oxidant properties and have been widely used in the skin care industry either as topically applied agents or oral supplements in an attempt to prolong youthful skin appearance. This review will provide an overview of the current literature "linking" nutrition with skin aging.
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Pistachio (Pistacia vera L.) nuts are a rich source of phenolic compounds, known for their high antioxidant activity, and contained not only in the seeds but also in the skin. A pistachio cultivar of high quality is typical of Bronte, Sicily, Italy. The purpose of our study was to investigate the chemical composition and antioxidant properties of two polyphenol-rich extracts from skins (TP) and decorticated seeds (SP) of Bronte pistachios, and to verify the potential use of these extracts for topical photoprotective products. Chemical analysis showed that the TP and SP extracts contain high levels of phenolic compounds, but the TP extract is about ten times richer in phenols than the SP extract, being anthocyanins the most abundant compounds found in the TP extract. Both these extracts, and especially the TP extract, possess good radical scavenger/antioxidant properties, as shown in a series of in vitro assays carried out using homogenous and non-homogenous chemical environment. Furthermore both the TP extract and, although at a lower degree, the SP extract reduce, when topically applied, UV-B-induced skin erythema in human volunteers. These findings suggest that extracts from Bronte TP and SP could be successfully employed as photoprotective ingredients in topical cosmetic and pharmaceutical formulations.
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The present study examines the effect of polyphenols extract of adlay (Coix lachryma-jobi L. var. ma-yuen Stapf) (APE) on high cholesterol diet fed rats (HCD). APE was orally administrated by gavage at doses of 10, 40 and 200 mg total phenolics/kg body weight of rats once a day for 28 days. At the end of four weeks, serum triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C), and markers of oxidative stress viz., malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) in the serum and liver of HCD and normal rats were assessed and compared. The results showed that administration of APE was significantly effective in decreasing the serum levels of TC, LDL-C and MDA, increasing the serum level of HDL-C and antioxidant capacity. In addition, oral gavage of APE could also increase the antioxidant capacity, CAT and GSH-Px activities in liver. These results suggested that APE exerted a high hypocholesterolemic and antioxidant activities, which might be characterized by a protective effect on cardiovascular health in vivo.
At a simplistic level, colorectal cancer arises from mutations in various proto-oncogenes and tumor suppressor genes. Aside from genetically inherited factors, environmental, lifestyle, and dietary habits have all been identified as risk agents promoting mutational events leading to the development of colorectal cancer. This "In Focus" presents evidence that the intestinal endogenous bacterial community represents a risk factor for the development of colorectal cancer. Cancer Discov; 3(4); 384-7. ©2013 AACR.
The composition of the human gut microbiota is related to host health, and it is thought that dietary habits may play a role in shaping this composition. Here, we examined the population size and prevalence of six predominant bacterial genera and the species compositions of genus Bifidobacterium (g-Bifid) and Bacteroides fragilis group (g-Bfra) in 42 healthy Belgian adults by quantitative PCR (qPCR) over a period of one month. The population sizes and prevalence of these bacteria were basically stable throughout the study period. The predominant g-Bifid species were Bifidobacterium adolescentis and Bifidobacterium longum ss. longum, and the predominant g-Bfra species were Bacteroides vulgatus, Bacteroides uniformis, and Bacteroidesovatus. The Belgian gut microbiota data were then compared with gut microbiota data from 46 Japanese subjects collected according to the same protocol (Matsuki et al., Appl. Environ. Microbiol. 70, 167-173,2004). The population size and prevalence of Bifidobacterium catenulatum group were significantly lower in the Belgian gut microbiota than in the Japanese gut microbiota (P < 0.001); however, the population size and prevalence of g-Bifid did not differ. This species-level qPCR analysis will be helpful for investigating the diversity of gut microbiota among ethnic groups.
Gut microbiota is a compilation of microorganisms dwelling in the entire mammalian gastrointestinal tract. They display a symbiotic relationship with the host contributing to its intestinal health and disease. Even a slight fluctuation in this equipoise may be deleterious to the host, leading to many pathological conditions like Clostridium difficile infection or inflammatory bowel disease (IBD). In this review, we focus on the role of microbial dysbiosis in initiation of C. difficile infection and IBD, and we also touch upon the role of specific pathogens, particularly C. difficile, as causative agents of IBD. We also discuss the molecular mechanisms activated by C. difficile that contribute to the development and exacerbation of gastrointestinal disorders.
Probiotics have been considered to affect not only the gut but also the skin. This study aimed at examining whether oral administration of live Bifidobacterium breve strain Yakult (BBY), a typical probiotic, could exert photoprotective effects in hairless mouse skin. BBY cell suspensions and fermented milk containing BBY (BBYM) were orally administered to hairless mice for 9 and 14 days, respectively. Mice were irradiated with ultraviolet (UV) light daily for the last four consecutive days. Twenty-four hours after the final irradiation, skin elasticity, appearance, elastase activity and interleukin (IL)-1β levels were evaluated in the dorsal skin. BBY and BBYM significantly prevented UV-induced deleterious changes in skin elasticity and appearance. BBY suppressed the increases in both elastase activity and IL-1β levels in the skin. There was a significant negative correlation between elastase activity and the ratio of elastic recovery to total deformation and a significant positive correlation between elastase activity and the area ratio of furrows, independent of UV irradiation or BBY administration. Our findings suggest that oral administration of probiotic BBY has the potential to prevent UV-induced skin damage, supporting the hypothesis that probiotics are beneficial not only to the intestine but also to the skin.
Antioxidants are molecules capable of inhibiting the oxidation of other molecules. Although oxidation reactions are essential for life, they can also be damaging. All living organisms maintain complex systems of multiple types of antioxidants to protect their cells from oxidative damage. Antioxidants can also act as pro-oxidants, under certain circumstances. The efficacy and benefit of an antioxidant is, therefore, very much dependent on the delivery of the antioxidant to the organism. Topically applied antioxidants constitute an important group of pharmacologically active agents capable of preventing the occurrence and reducing the severity of UV-induced skin damage and skin aging. Antioxidants protect skin cells against the damaging effects of reactive oxygen species (ROS), such as singlet oxygen, superoxide, peroxyl radicals, hydroxyl radicals, and peroxynitrite. ROS induced oxidative stress in the skin has been linked to cancer, aging, inflammation, and photodamage. This review focuses on antioxidants used in the cosmetic industry for protection of skin, formulation methods used to enhance their efficacy, and methods used to test the efficacy of antioxidants in topical formulations.