Biol 124

10/22/14
The Effects of Circadian Rhythm on Cardiac Variation Between Genders
Introduction:
Circadian rhythm is the endogenous but entrainable oscillation of regular biological
processes and functions around a ~24 hour period. Circadian rhythm is often thought of as a
biological clock that dictates the activity level of various biological functions such as brain
activity, core body temperature, hormone regulation, metabolism, and other autonomous
processes. In this lab, we are interested in the effects of circadian rhythm on resting heart rate
throughout normal waking periods. From previous studies, it has been shown that circadian
rhythm does affect resting heart rate throughout the day as the body goes through various
changes in response to hormones and other stimuli during a circadian cycle (Massin). However,
they did not study how resting heart rate changed in response to circadian rhythm over the course
of a day and whether this change was universal or distinctly different between males and
females. This is an important distinction to make because one of the leading causes of death for
both males and females is cardiovascular disease which is the complicated result of a myriad of
contributing factors ranging anywhere from gender, genetics, lifestyle, and more. In particular,
cardiovascular disease tends to affect more women than men, claiming about 40% of all deaths
in American women1. Therefore, in this experiment we wish to see if there is a difference
between the natural variation in heart rate due to the body's own circadian rhythm between men
and women that could explain why women are more likely to suffer from cardiovascular disease.
After going through the first half of Biol-124 lab on animal anatomy and physiology, this
seemed like a novel experiment as it could potentially provide a significant answer to why one
gender is inherently more at risk than another for cardiovascular disease. Not only that, but the
experiment provides an opportunity to use the skills and techniques learned in lab outside of the
laboratory. The cardiac function lab was interesting conceptually, but was also severely limited
as the entire experiment had to completed within a 2-3 hour lab time. This time constraint
restricted the variety and novelty of experiments that can be performed in that limited window to
those that were either very obvious or very mundane. Therefore, this experiment allows us to
apply the theory learned to a more complex experiment that instead of spanning 2-3 hours, spans
2-3 days.
I find this question personally fascinating because circadian rhythm in itself is an
interesting concept but also because as college students, we are not exactly known for having
regular sleep schedules. Therefore, it would be interesting to see how my sleep schedule and by
extension circadian rhythm affects my cardiovascular health.
Objective: The objective of this experiment is to determine the effects of circadian rhythm on
cardiac variations between male and female genders.

Research Hypothesis: If circadian variation in heart rate is influenced by gender, then there will
be a greater difference in women's circadian variation in heart rate than in men.
Null Hypothesis: There is no difference in circadian variation between men and women.
Alterative Hypothesis: There is a statistically significant difference between circadian variation
between men and women.
Procedure:
Two groups of 9 subjects, 9 male and 9 female, will take their heart rates by measuring
their pulse while sitting down for 30 seconds and then multiplying by two. They will take these
heart rates at set intervals throughout the day: 9:00 am, 10:00 am, 12:00 pm, 5:00 pm and 10:00
pm and record them for 2 to 3 days, where each additional day is an iteration. The times chosen
correspond to key circadian events such as the release of testosterone, melatonin, or correspond
to periods of highest cardiovascular efficiency among other things that may influence heart rate2.
Graphs and Data:
Males:
Table 1: Heart Rate Data for Subject M1:

Time

9:00
AM
10:00
AM
12:00
PM
5:00
PM
10:00
PM

Heart
Rate
(Day
1)
74

Heart
Rate
(Day
2)
56

Heart
Rate
(Day
3)
56

75

68

66

66

76

64

69

72

74

76

76

64

Averag
e Heart
Ratet

Differen
cev

62

-6.8

69.666
67
68.666
67
71.666
67
72

0.86666
7
0.13333
2.86666
7
3.2

*Heart Rate is measured in BPM (beats/min)

t
v

The average heart rate here is the average of the heart rates taken at the same time across all days-applies for all individual tables below

The difference here is calculated by subtracting the average heart beat rate of that individual in throughout a day by the average heart rate for
that time. -applies for all individual tables below.

Table 2: Heart Rate Data for Subject M2:

Time

9:00
AM
10:00
AM
12:00
PM
5:00
PM
10:00
PM

Heart
Rate
(Day
1)
75

Heart
Rate
(Day
2)
75

Heart
Rate
(Day
3)
75

Avera
ge
Heart
Rate
75

Differen
ce

72

78

72

74

72

81

75

76

72

75

75

74

78

66

69

71

-3

Averag
e Heart
Rate

Differen
ce

69.666
67
67.333
33
66

4.13336
7
1.80003
3
0.4667

63.333
33
61.333
33

2.19997
4.19997

Averag
e Heart
Rate

Differen
ce

66.666
67
69

3.20003
-0.8667

*Heart Rate is measured in BPM (beats/min)

Table 3:Heart Rate Data for Subject M3:

Time

9:00
AM
10:00
AM
12:00
PM
5:00
PM
10:00
PM

Heart
Rate
(Day
1)
78

Heart
Rate
(Day
2)
66

Heart
Rate
(Day
3)
65

68

66

68

56

74

68

48

71

71

52

68

64

*Heart Rate is measured in BPM (beats/min)

Table 4:Heart Rate Data for Subject M4:

Time

9:00
AM
10:00

Heart
Rate
(Day
1)
66
72

Heart
Rate
(Day
2)
68
70

Heart
Rate
(Day
3)
66
65

AM
12:00
PM
5:00
PM
10:00
PM

74

70

72

72

2.1333

72

70

70

72

69

72

70.666
67
71

0.79996
7
1.1333

Averag
e Heart
Rate

Differen
ce

62

-5.8667

75.333
33
64.666
67
66

7.46663
3
3.20003
-1.8667

71.333
33

3.46663
3

Averag
e Heart
Rate

Differen
ce

54

11.8667
9.20003
9.46663
3
4.79996
7
6.79996
7

*Heart Rate is measured in BPM (beats/min)

Table 5:Heart Rate Data for Subject M5:

Time

9:00
AM
10:00
AM
12:00
PM
5:00
PM
10:00
PM

Heart
Rate
(Day
1)
58

Heart
Rate
(Day
2)
58

Heart
Rate
(Day
3)
70

66

88

72

62

64

68

70

60

68

60

78

76

*Heart Rate is measured in BPM (beats/min)

Table 6: Heart Rate Data for Subject M6:

Time

9:00
AM
10:00
AM
12:00
PM
5:00
PM
10:00
PM

Heart
Rate
(Day
1)
54

Heart
Rate
(Day
2)
52

Heart
Rate
(Day
3)
56

60

58

52

76

76

74

58

94

60

84

64

70

*Heart Rate is measured in BPM (beats/min)

56.666
67
75.333
33
70.666
67
72.666
67

Table 7: Heart Rate Data for Subject M7:

Time

9:00
AM
10:00
AM
12:00
PM
5:00
PM
10:00
PM

Heart
Rate
(Day
1)
62

Heart
Rate
(Day
2)
58

Heart
Rate
(Day
3)
60

Averag
e Heart
Rate

Differen
ce

60

-6.8667

67

66

68

67

0.1333

68

68

70

74

75

77

65

63

62

68.666
67
75.333
33
63.333
33

1.79996
7
8.46663
3
3.53337

*Heart Rate is measured in BPM (beats/min)

Table 8: Heart Rate Data for Subject M8:

Time

9:00
AM
10:00
AM
12:00
PM
5:00
PM
10:00
PM

Heart
Rate
(Day
1)
64

Heart
Rate
(Day
2)
50

Heart
Rate
(Day
3)
X

Avera
ge
Heart
Rate
57

Differen
ce

-10.8

64

52

58

-9.8

72

72

72

4.2

80

76

78

10.2

84

64

74

6.2

Avera
ge
Heart
Rate
75

Differen
ce

Note: X means no data was recorded by subject

*Heart Rate is measured in BPM (beats/min)

Table 9: Heart Rate Data for Subject M9:

Time

9:00

Heart
Rate
(Day
1)
68

Heart
Rate
(Day
2)
82

Heart
Rate
(Day
3)
X

-1

AM
10:00
AM
12:00
PM
5:00
PM
10:00
PM

70

78

74

-2

80

80

80

76

76

76

74

76

75

-1

Averag
e Heart
Rate

Differen
ce

78.666
67
85.333
33
88

1.60003
5.06663
3
7.7333

81.333
33
68

1.06663
3
12.2667

Averag
e Heart
Rate

Differen
ce

5.80003
2.80003
4.86663
3
3.5333
-9.4667

Note: X means no data was recorded by subject

*Heart Rate is measured in BPM (beats/min)

Females:
Table 10: Heart Rate Data for Subject F1:

Time

9:00
AM
10:00
AM
12:00
PM
5:00
PM
10:00
PM

Heart
Rate
(Day
1)
68

Heart
Rate
(Day
2)
96

Heart
Rate
(Day
3)
72

92

84

80

80

100

84

76

80

88

72

68

64

*Heart Rate is measured in BPM (beats/min)

Table 11: Heart Rate Data for Subject F2:

Time

9:00
AM
10:00
AM
12:00
PM
5:00
PM
10:00
PM

Heart
Rate
(Day
1)
59

Heart
Rate
(Day
2)
58

Heart
Rate
(Day
3)
71

72

60

65

60

89

71

56

97

63

62.666
67
65.666
67
73.333
33
72

55

62

60

59

*Heart Rate is measured in BPM (beats/min)

Table 12: Heart Rate Data for Subject F3:

Time

9:00
AM
10:00
AM
12:00
PM
5:00
PM
10:00
PM

Heart
Rate
(Day
1)
58

Heart
Rate
(Day
2)
54

Heart
Rate
(Day
3)
52

Averag
e Heart
Rate

Differen
ce

3.33333
2.66666
7
0.66666
7
0

62

56

64

62

60

54

60

58

56

54.666
67
60.666
67
58.666
67
58

60

60

54

58

Averag
e Heart
Rate

Differen
ce

50

-3.0667

49.666
67
55

3.40003
1.9333

56.333
33
54.333
33

3.26663
3
1.26663
3

*Heart Rate is measured in BPM (beats/min)

Table 13: Heart Rate Data for Subject F4:

Time

9:00
AM
10:00
AM
12:00
PM
5:00
PM
10:00
PM

Heart
Rate
(Day
1)
52

Heart
Rate
(Day
2)
48

Heart
Rate
(Day
3)
50

47

50

52

55

56

54

57

58

54

57

52

54

*Heart Rate is measured in BPM (beats/min)

Table 14: Heart Rate Data for Subject F5:

Time

9:00
AM
10:00

Heart
Rate
(Day
1)
68
70

Heart
Rate
(Day
2)
70
68

Heart
Rate
(Day
3)
X
X

Avera
ge
Heart
Rate
69

Differen
ce

69

-3.6

-3.6

AM
12:00
PM
5:00
PM
10:00
PM

76

78

77

4.4

70

76

73

0.4

70

80

75

2.4

Avera
ge
Heart
Rate
77

Differen
ce

Note: X means no data was recorded by subject

*Heart Rate is measured in BPM (beats/min)

Table 15: Heart Rate Data for Subject F6:

Time

9:00
AM
10:00
AM
12:00
PM
5:00
PM
10:00
PM

Heart
Rate
(Day
1)
74

Heart
Rate
(Day
2)
80

Heart
Rate
(Day
3)
X

-5.9

75

78

76.5

-6.4

90

86

88

5.1

84

88

86

3.1

98

76

87

4.1

Averag
e Heart
Rate

Differen
ce

76.666
67
84.666
67
81.333
33
82

2.53333
5.46666
7
2.13333
3
2.8

71.333
33

7.86667

Note: X means no data was recorded by subject

*Heart Rate is measured in BPM (beats/min)

Table 16: Heart Rate Data for Subject F7:

Time

9:00
AM
10:00
AM
12:00
PM
5:00
PM
10:00
PM

Heart
Rate
(Day
1)
84

Heart
Rate
(Day
2)
74

Heart
Rate
(Day
3)
72

82

100

72

84

78

82

70

86

90

66

72

76

*Heart Rate is measured in BPM (beats/min)

Table 17: Heart Rate Data for Subject F8:

Time

9:00
AM
10:00
AM
12:00
PM
5:00
PM
10:00
PM

Heart
Rate
(Day
1)
70

Heart
Rate
(Day
2)
72

Heart
Rate
(Day
3)
68

76

74

74

86

88

80

78

76

76

74

70

74

Averag
e Heart
Rate

Differen
ce

70

-5.733

74.666
67
84.666
67
76.666
67
72.666
67

1.06633
8.93366
7
0.93366
7
3.06633

*Heart Rate is measured in BPM (beats/min)

Table 18: Heart Rate Data for Subject F9:

Time

9:00
AM
10:00
AM
12:00
PM
5:00
PM
10:00
PM

Heart
Rate
(Day
1)
66

Heart
Rate
(Day
2)
60

Heart
Rate
(Day
3)
X

Avera
ge
Heart
Rate
63

Differen
ce

-14.5

72

72

72

-5.5

90

84

87

9.5

85

84

84.5

84

76

80

2.5

Note: X means no data was recorded by subject

*Heart Rate is measured in BPM (beats/min)

Joint Data for Comparison:

Table 19: Average Heart Rate and SD throughout the day for Male and Female Subjects

Subjec
t

Avera
ge
Heart
Rate

M1
M2
M3

68.8
74
65.33

SD of
Avera
ge
Heart
Rate
4.04
1.87
3.28

Subjec
t

Avera
ge
Heart
Rate

F1
F2
F3

80.27
66.53
58.0

SD of
Avera
ge
Heart
Rate
7.74
6.09
2.16

M4
M5
M6
M7
M8
M9
All
Males

69.86
7
67.87
65.87
66.86
67.8
76
69.18

2.09

F4

53.07

3.04

5.38
9.80
5.80
9.65
2.35
2.98

F5
F6
F7
F8
F9
All
Femal
es

72.6
82.9
79.2
75.73
77.3
71.73

3.58
5.66
5.26
5.57
9.82
4.02

*Heart Rate is measured in BPM (beats/min)

Table 20: Average Heart Rate, Difference in HR and SD at various set times for Males and Females

Time

9:00
AM
10:00
AM
12:00
PM
5:00
PM
10:00
PM
SD

Average
Difference from
Mean HR for All
Males
-4.585

Average HR for
all Males

-1.289

67.889

2.303

71.481

2.563

71.741

1.007

70.185

3.05

2.98

Average
Difference from
Mean HR for All
Females

Average HR for
all Females

-5.118

66.852

-1.063

70.907

5.029

77.0

2.456

74.426

-2.489
2.372

69.482
4.02

64.593

*Heart Rate is measured in BPM (beats/min)

Fig 1: Graph of the average heart rate of 9 men at 5 time intervals

Average HR Throughout the Day for 9 Men

74
72
70
68
66
64
62
60
Heart Rate (Beats/Min)

Time

Average Difference in HR Throughout the Day for 9 Men

Heart Rate (Beats/Min)

3
2
1
0
-1
-2
-3
-4
-5
Time

Fig 2: Graph of the average difference in heart rate of 9 men at 5 time intervals

Average HR Throughout the Day for 9 Women

80
75
70
65
60
Heart Rate (Beats/Min)

Time

Fig 3: Graph of average heart rate of 9 women at 5 time intervals

Average Difference in HR Throughout the Day for 9 Women

6
4
2
Heart Rate (Beats/Min)

0
-2
-4
-6

Time

Fig 4: Graph of average difference in heart rate of 9 women at 5 time intervals

Trends:
Although no clear trends in mean heart rate and heart rate difference can be seen from
individual data of male and female subjects, if the values from the two groups are averaged, we
then begin to see something potentially significant. From the data we see that females tend to
have a slightly higher resting heart rate than males. Not only that, but also the standard deviation
in mean heart rate is also higher for females than males.
From the graphs of average heart rate plotted against the 5 time intervals (Fig 1 &3), we
are unable to see any distinct difference between male and female variation in circadian rhythm

other than that for both groups heart rate starts lowest in the morning upon waking up and
increases during the day until it falls again at night before sleeping. However, when looking at
the graphs of average difference in heart rate throughout the day (Fig 2 &4), the differences in
heart rate due to circadian variation become more clear. For example, there is only a moderate
increase in heart rate above the average at noon for males while female exhibited almost an
average increase of almost twice as much. Not only that but, in males the heart rate never
appears to decrease below the average heart rate other than in the morning upon first waking up.
This is not seen in females where both upon waking up and right before going to sleep that the
heart rate is seen going below the average heart rate. Finally, the observed peak difference in
heart rate is highest for males at 5:00 pm where it is noon for females.
Unpaired t-Test:

Table 21: Relevant Data for unpaired two tailed t stat test

Males
Mean change throughout day
SD
n
t
Two tailed P value
Critical value

Females
2.35
2.98
9

3.231
4.02
9
.5282
.6056
2.1199

*Values obtained from stat package after entering means, SD, and n

Discussion:
From the values obtained after doing a variation of the unpaired two tailed t test that
assumes different variances, it is found that the p valued obtained of .6056 which is less than the
critical value of 2.1199 with 16 degrees of freedom at a .05 false positive rate. This means that

there is not a statistically significant difference in circadian variation in heart rate due to gender.
Therefore, we do not have enough evidence to reject our null hypothesis.
Our initial prediction from reading scientific literature on subject was that there would be
a difference in circadian variation in heart rate, more specifically that females would have greater
variation. This is because circadian rhythm influences the activity level various hormones during
the day, some of which, like testosterone, are gender biased. Therefore, it would make sense that
circadian rhythms should differ between genders and by extension circadian variation in heart
rate because it was established in prior literature that resting heart rate is indeed influenced by
circadian rhythm.
Upon first glance the results were as we expected as we obtained a higher average resting
heart rate among the females than the males. Not only that but the variation in heart rate during
the course of a day was also higher in females than males. The first part of our prediction that
circadian variation in heart rate was different between the genders was supported by figures 2
and 4 where one sees the average deviations in heart rate at different times of the day from the
two genders. The trends and patterns of these two graphs look significantly different (differences
noted above under Trends) and thus reaffirmed our initial suspicions. Furthermore, the data also
appeared to support our premonition that variation in females would be greater because the
average difference in heart rate for their peak at noon was double that of the male group. Not
only that but we predicted that 5:00 pm would give the highest heart rate because that is when
cardiovascular efficiency is greatest, and we know from the cardiac function lab that muscle
contractions do affect heart rate positively. This was seen in males but not in females, probably
because the muscle mass in males is greater. Not only that but. we also predicted that heart rate
would decrease at 10:00 pm because that is when melatonin secretion begins which is associated

with sleep and thus should correlate to a decrease in heart rate which was observed in both
genders. Unfortunately, after doing an unpaired two tailed t stat test, it was found that the
variation and results obtained were not statistically significant with a .05 false positive rate or
less.
However, this does not mean that there is no correlation at all because looking at the data
it seems unlikely that there is no correlation at all. This inability to arrive at a definite conclusion
could be due to dozens of other biological and external factors (physical activity, stress,
environment, and more) that could have been influencing the heart rates of test subjects during
the course of the experiment. Therefore, it is an error on the side of experimental design. Ideally,
to minimize any and all possible variations other than circadian variation due to gender we
would place all out test subjects in a controlled environment for the course of the experiment
where they just remain stationary the entire time and all follow the same routine with the same
waking time, sleeping time, eating time, and so forth. This, although unfeasible, would allow us
to get the most error free data in comparison to allowing the subjects to go about their day where
external stimuli and different schedules are all influencing the results. In fact, as mentioned in the
very beginning, circadian rhythm is entrainable which means that cycles will vary depending on
the person and their lifestyle. Therefore, variations exist not only between genders but also
between individuals within the same gender.
As stated in the introduction, this experiment is both novel and noble a quick search of
PubMed and Google scholar reveal no results detailing the relationship between circadian
variation in heart rate and gender. Therefore, we are unable to compare our results to those done
by other scientists. However, a related relevant question to ask after completing the experiment
are how does circadian rhythm differ between the two genders exactly? The answer to this

question would help to decide in the future, what would be the best times to record heart rate
because choosing times that match key gender specific/biased events in circadian rhythm would
allow us to get a clearer picture of the difference in heart rate due to circadian variation and
ignore times where circadian rhythm is the same and thus cardiac variation should be similar.

Works Cited
Baylor College of Medicine. "Heart beats to the rhythm of a circadian clock." ScienceDaily.
ScienceDaily, 22 February 2012.
<www.sciencedaily.com/releases/2012/02/120222132559.htm>.

Massin, M., Maeyns, K., Withofs, N., Ravet, F., Gerard, P., & Healy, M. (2000).
Circadian rhythm of heart rate and heart rate variability.
Archives Disease Childhood, 83, 179-182
Other useful references:
1

https://www.goredforwomen.org/about-heart-disease/facts_about_heart_disease_in_women-sub-category/statisticsat-a-glance/
2

http://upload.wikimedia.org/wikipedia/commons/thumb/3/30/Biological_clock_human.svg/1200pxBiological_clock_human.svg.png