Vol.5, No.11, 1133-1138 (2013) Natural Science
http://dx.doi.org/10.4236/ns.2013.511139
Can the early bird catch the worm?
Effects of early rising on leukocyte subsets via
modification of autonomic nervous system and the
effect on glucose levels
Mayumi Watanabe1,2*, Yiwei Ling1, Chikako Tomiyama3, Kiyoshi Adachi4,5, Hidetoshi Mori2,
Kazushi Nishijo2, Toru Abo5, Kohei Akazawa1
1Department of Medical Informatics, Niigata University Medical & Dental Hospital, Niigata, Japan;
*Corresponding Author:
2Department of Health, Faculty of Health Science, Tsukuba University of Technology, Tsukuba, Japan
3School of Health Sciences, Faculty of Medicine, Niigata University, Niigata, Japan
4Shinjuku Acupuncture and Judo Therapy School, Tokyo, Japan
5Department of Immunology, Niigata University School of Medicine, Niigata, Japan
Received 4 September 2013; revised 4 October 2013; accepted 11 October 2013
Copyright © 2013 Mayumi Watanabe et al. This is an open access article distributed under the Creative Commons Attribution Li-
cense, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
ABSTRACT
The importance of sleep has been described in
proverbs such as “the early bird catches the
worm”. However, there are few scientific reports
on the effects of early rising. Therefore, early
risers (Group E) and late risers (Group L) were
compared and the number and ratio of leuko-
cytes, body temperature, glucose and its asso-
ciated hormones were studied. Furthermore,
each group was divided into two groups by the
duration of sleep and the following four groups
were compared: early risers with short sleep
(Group E-S); early risers with long sleep (Group
E-L); late risers with short sleep (Group L-S);
and late risers w ith long sleep (Group L-L). Then,
compared with Group L, Group E showed lower
numbers and ratio of granulocytes and a higher
ratio of lymphocytes showing parasympathetic
nerve dominance. Group E sho wed higher levels
of glucose and its related hormones than Group
L, indicating sympathetic nerve dominance.
Compared with Groups E-S and L-S, Groups E-L
and L-L showed lower glucose and cortisol lev-
els, respectively. These results indicated that
early rising might affect leukocyte subsets, and
adequate duration of sleep could decrease lev-
els of glucose via modification of the autonomic
nervous system.
Keyw ords: Early Rising; Leukocyte; Autonomic
Nervous System (ANS)
1. INTRODUCTION
Sleep is an indispensable action and its importance has
been related in many proverbs such as “the early bird
catches the worm” or the Japanese saying “sleep brings
up a child well”. However, there are few scientific re-
ports that have investigated the influence of sleep on the
number and the ratio of leukocytes and the autonomic
nervous system (ANS). Adachi et al. reported that the
ratio of lymphocytes in persons who went to bed early
was higher than that of subjects who sleep late [1]; how-
ever, the effects of early rising were not reported.
Therefore, in this study 23 subjects were divided into
two groups, early risers (Group E) and late risers (Group
L), and the number and ratio of leukocytes, body tem-
perature, glucose, and glucose-associated hormones [cor-
tisol (CR), adrenocorticotropic hormone (ACTH), no-
radrenaline (NA) and growth hormone (GH)] were inves-
tigated. Furthermore, each group was divided into two groups
to create the following four groups: early risers with short
sleep (Group E-S); early risers with long sleep (Group E-L);
late risers with short sleep (Group L-S); and late risers
with long sleep (Group L-L). Through the comparison of
these groups, the efficiency of early rising was revealed.
2. MATERIALS AND METHODS
2.1. Subjects
Twenty-three healthy volunteers (ages 20 to 59; avera-

M. Watanabe et al. / Natural Science 5 (2013) 1133-1138
1134
ge 38.1 ± 13.5 years old) participated in this study. Writ-
ten informed consent was obtained from all subjects and
the study was approved by the institutional review board
of Niigata University.
2.2. Early Risers (Group E) and Late Risers
(Group L)
Subjects were divided into 2 groups by the time of
their rising. Group E, the group of early risers, had 12
members who regularly rose before 8:00 am (including
8:00 am). Group L, the group of late risers, had 11 sub-
jects who usually rose after 8:00 am (excluding 8:00 am).
Group E was divided into the following two groups at
the border of 6.20 hours: early risers with short sleep
group (Group E-S); and early risers with long sleep
group (Group E-L). In the same way, Group L was di-
vided into the late risers with short sleep group (Group
L-S) and the late risers with long sleep group (Group
L-L). Details of the subjects are shown in Table 1.
2.3. Venous Blood Samples and Analysis
As mentioned in a previous study [1], fresh venous
blood (25 mL) for analysis was obtained from the fore-
arm median antebrachial vein. Body temperature was
measured under the arm (axillary). Considering the ef-
fects of circadian rhythm and nutrition intake, these data
were obtained between 16:30 and 17:30, more than 4
hours after eating. Leukocyte subsets were determined
using a hemocytometer and the May-Grünwald Giemsa
stain method. Blood glucose levels were measured by
Precision Xtra TM (Abott Japan Co., Ltd., Chiba, Japan).
Blood samples were sent to Mitsubishi Chemical Medi-
ence Corporation (Tokyo, Japan) for the analysis of CR,
ACTH, GH and NA. The levels of CR, NA and ACTH
and of GH were analyzed by radioimmunoassay (coated
tube solid phase method), high performance liquid
chromatography and immunoradiometric assay (beads
solid phase method), respectively.
2.4. Statistical Analysis
All analyses were performed using the Statistical
Package for the Social Sciences (SPSS) Version 20. The
difference between values was determined by Student’s
t-test, Mann-Whitney’s U test, Welch’s t-test between
two groups comparison (Group E vs. Group L) and
Kruskal-Wallis test among four groups comparison
(Group E-S, Group E-L, Group L-S and Group L-L).
Figures are all shown as the mean ± one S.D. P-values <
0.05 were considered to indicate statistical significance,
and all statistical tests were two-tailed.
3. RESULTS
3.1. Age, Gl ucose and Body Temperature
To study the effects of early rising, the average ages,
glucose levels and body temperatures of subjects were
compared between two groups. The average age of
Group E (45.1 ± 12.0) was significantly older than that of
Group L (30.5 ± 10.9) (P < 0.05), and the level of glu-
cose of the former (95.1 ± 7.9 mg/dL) was remarkably
higher than that of the latter (86.7 ± 10.4 mg/dL) (P <
0.05). The average of both groups stayed within the
normal range. Subjects with hypothermia (<36.0˚C) were
found only in Group L; however, there was no significant
difference between the two groups in terms of body
temperature (Figure 1).
3.2. The Number and the Ratio of
Leukocytes
The number of granulocytes of Group E subjects was
statistically lower than that of Group L subjects (3.3 ±
0.9 vs. 4.2 ± 1.3 × 103/μL) (P < 0.05). The number of
lymphocytes of subjects in Group E tended to be higher
than those in Group L; however, there was no significant
difference (2.5 ± 0.6 vs. 2.1 ± 0.6 × 103/μL). The ratio of
granulocytes of Group E was lower than that of Group L
(54.0% ± 8.3% vs. 62.5% ± 7.7%) (P < 0.05), while the
Table 1. Details of subjects. Group E (early risers: rising up before 8:00 am, including 8:00 am), Group L (late risers, rising up after 8:00 am, exclud-
ing 8:00 am), Group E-S (early risers with short sleep), Group E-L (early risers with long sleep),Group L-S (late risers with short sleep), Group L-L
(late risers with long sleep).
n (m, f) Age Time to sleep Time to rise up Duration (H)
Group E 12 (7, 5) 45.1 ± 12.0 23:50 ± 0:51 6:05 ± 1:05 6:15 ± 0:57
Group ES 8 (5, 3) 45.1 ± 13.3 23:52 ± 1:01 5:33 ± 0:54 5:41 ± 0:27
Group EL 4 (2, 2) 45.0 ± 11.0 23:45 ± 0:30 7:07 ± 0:28 7:22 ± 0:37
Group L 11 (7, 4) 30.5 ± 10.9 27:16 ± 1:07 9:35 ± 0:55 6:19 ± 0:58
Group LS 5 (3, 2) 30.4 ± 12.2 27:54 ± 1:14 9:18 ± 0:39 5:24 ± 0:37
Group LL 6 (4, 2) 30.5 ± 10.9 26:45 ± 0:45 9:50 ± 0:45 7:05 ± 0:12

M. Watanabe et al. / Natural Science 5 (2013) 1133-1138 1135
Figure 1. Age, glucose and
body temperature. The aver-
age age of Group E was sig-
nificantly older than that of
Group L (P < 0.05), and the
levels of glucose of the for-
mer were remarkably higher
than those of the latter (P <
0.05). Subjects with hypo-
thermia (<36.0˚C) were found
only in Group L; however,
there was no significant dif-
ference between the two
groups.
ratio of lymphocytes was precisely the opposite (40.9% ±
8.5% vs. 31.4% ± 7.2%) (P < 0.05) (Figure 2).
3.3. CR, ACTH, NA and GH
There was a lot of individual variation among hor-
mones, showing no significant differences between the
two groups; however, subjects with high levels (over the
normal range) of CR and GH were found only in Group
E. The levels of ACTH in Group E subjects were higher
than that of Group L, though without a significant dif-
ference (27.1 ± 19.0 vs. 21.4 ± 7.7 pg/mL). The levels of
NA in both groups were roughly the same (Figur e 3).
Figure 2. The number and ratio of leukocytes in
venous blood. The number of leukocytes and
granulocytes of Group E were statistically lower
than that of Group L (P < 0.05). The number of
lymphocytes in Group E tended to be higher;
however, there were no significant differences.
The ratio of granulocytes of Group E was lower
than that of Group L (P < 0.05), while the ratio
of lymphocytes was precisely the opposite (P <
0.05).
3.4. Preparation of Age Matched Groups
As described above, the average age of Group E was
older than that of Group L. To avoid the possibility of the
effects of aging [2,3] and the duration of sleep [4-6],
each group was divided again and then the duration of
sleep (6.20 hours) and age-matched four groups were
prepared (Table 1).
3.5. Comparison among Four
Groups-Glucose, CR and GH
The levels of glucose were as follows: Group ES (96.3
± 6.6 mg/dL) > Group E-L (92.8 ± 10.8 mg/dL) > Group
L-S (90.4 ± 11.1 mg/dL) > Group L-L (83.7 ± 9.7
mg/dL). Levels of CR in Group E-S (11.7 ± 5.2 μg/dL)
were higher than that of Group E-L (9.4 ± 4.3 μg/dL). In
the same way, levels of CR in Group L-S (12.6 ± 6.3
μg/dL) were higher than Group L-L (11.3 ± 3.7 μg/dL),

M. Watanabe et al. / Natural Science 5 (2013) 1133-1138
1136
Figure 3. The plasma levels of various hormones.
There was a lot of individual variation in the levels
of hormones, with no significant differences. High
levels of CR, ACTH, NA and GH were observed
only in Group E. The dotted lines in the figures in-
dicate normal levels.
without a significant difference. High levels of GH were
shown only in the groups with longer sleep duration
(1.21 and 2.04 ng/dL) (Figure 4).
4. DISCUSSION
4.1. Leucocyte Subsets and Body
Temperature Indicated That Early Rising
Induced PN Dominance
(SN Suppression)
In this study, the effect of early rising was revealed, as
well as that of going to bed early [1]. In fact, compared
with Group L, Group E showed a lower number and ratio
of granulocytes and a higher ratio of lymphocytes. The
average age of Group E was older than that of Group L.
It has been reported that aging inducing sympathetic
nerve (SN) dominance, with a subsequent increase in
granulocytes and decrease in lymphocytes [7-9]. The
results of this study indicated Group E showed more
parasympathetic nerve (PN) dominance (SN suppression)
than Group L based on leukocyte subset levels. This hy-
pothesis also was supported by body temperature be-
cause it has been reported that hypothermia can be in-
duced by SN dominance [10,11].
4.2. Glucose Levels Were Higher in Early
Risers than Late Risers
The levels of glucose of subjects in Group E were sig-
nificantly higher than those in Group L. One reason
Figure 4. Comparison among four
groups: glucose, CR and GH. The levels
of glucose were as follows: Group E-S
> Group E-L > Group L-S > Group L-L.
could be that early rising induces a mild SN dominance
and it has been reported that SN dominance elevates the
level of glucose [10,11]. This finding conflicts with the
above mentioned hypothesis concerning subsets of leu-
kocyte. The difference in the average age between the
two groups must be considered (45.1 ± 12. 0 vs. 30.5 ±
10.8) because it is well known that elevated levels of
glucose are directly affected by aging [2,3]. An associa-
tion with the duration of sleep also was reported [4-6].
Therefore, it was required to evaluate the difference.
4.3. Sleep Lowered Glucose Levels via PN
Dominance (SN Suppression)
The average age and sleep duration were matched be-
tween Groups E-S and E-L, and between Groups L-S and
L-L (Table 1). This was because Group E-S showed
higher levels of glucose and CR than Group E-L (Figure
4). In the same way, levels in Group L-S were higher
than those of Group L-L. In short, it was understood that
compared with short sleep (4.30 - 6.20 hours), long sleep

M. Watanabe et al. / Natural Science 5 (2013) 1133-1138 1137
(6.20 - 8.00 hours) decreased the levels of glucose, irre-
spective of age and the time of rising. In the age-
matched groups of this study the results agreed with
former reports [4-6]. It was considered that sleep induced
PN dominance (SN suppression) [12], which has been
reported to decrease glucose levels [11]. However, fur-
ther study in large-scale group is needed to investigate
the possibility that early rising elevates the level of glu-
cose via a mild SN dominance.
4.4. Early Rising with Adequate Sleep
Duration Might Provide the Anti-Aging
Effects via Modification of ANS Balance
and Leukocyte Subsets.
It is well known the sleep curtailment has become a
common behavior in modern society [13]. In modern
society, there are many occupations that need to keep
irregular hours with such efforts, such as doctors, nurses
and healthcare workers [14,15]. The results of this study
might provide a clue for their health promotion. Early
rising sometimes requires efforts and strong will, and
there is a possibility that it might induce a mild increase
of glucose levels within the normal range. Therefore,
maintaining an adequate sleep duration is recommended
because it could suppress the possibility of SN domi-
nance and its associated hormones such as CR, ACTH
and NA, which were often induced by aging. In short,
early rising with adequate sleep could be expected as
anti-aging effects. At the same time, the possibility of an
increase of GH, which is known to regulate carbohydrate
metabolism and promote anabolism and lipolysis [16],
must be studied in the future.
5. CONCLUSION
The early bird can catch the worm with the effects of
anti-aging because in this study it revealed that early
rising might affect leukocyte subsets and adequate dura-
tion of sleep and could decrease levels of glucose via
modification of the ANS. To maintain adequate sleep
duration, going to bed early was needed. In cases in
which persons stayed up late, efforts and ingenuity to get
additional longer sleep duration were necessary.
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