The Mean Testosterone Level is (verb should be capitalized) Higher in Adult Males With
(preposition should not be not capitalized) Deeper Voice
(Suggested title: Testosterone Levels Are Directly Related to the Deepness of Voice in Human
Males)
Ben Hoffman, Sid Mirgati, Tim Turnbaugh (space between title and name of authors)
Department of Biological Science
Saddleback College
Mission Viejo, CA 92692
Testosterone is an anabolic steroid hormone in males that has variety of responsibilities
including fetal development of genitalia and physical maturation during puberty; thus it
can be hypothesized that testosterone levels are directly related to the deepness of voice in
human males. To test this, (delete) a A Salimetrics kit was used to measure the amount of
free testosterone in subjects’ saliva… (Join sentences)…A micro-plate reader was used to
provide a quantified testosterone concentration reading. The voice and saliva of ten (n=10)
adult males was were collected. Normally Testosterone levels fluctuate throughout the
day, thus multiple saliva samples were collected and the mean value of the samples was
used (for analysis) in calculations (delete: calculations). One set of the lowest note a
subject could produce was also collected. After the analysis, (delete: After the analysis) the
mean voice frequency of all subjects was measured and the samples were divided into two
groups. The first group included the subjects with lower voice frequency than the total
average frequency. The second group included the subjects with higher voice frequency
than the total average frequency. The mean testosterone level of first group was 626.139
pg/mL (S.E.M ±134.728 pg/mL, C.L. ±428.766). The mean testosterone level of second
group was 475.896 pg/mL (S.E.M ±35.546 pg/mL, C.L. ±91.375). (correct the repetition of :
first group, second group) To compare the mean testosterone value of each group, a onetailed unpaired t-test was performed (P=0.179). The results rejected the hypothesis and
stated there is no significant correlation between the testosterone level and voice frequency
in adult males.
Introduction (it has to be centered)
Testosterone - a (an steroid and) key
hormone present in every man – (is
responsible for many functions and
developments in the human male) (delete).
It is responsible for fetal development of the
male genitalia, the physical changes that
occur during male puberty, and a variety of
other functions in the adult male, including
sperm production, erections, sex drive,
muscle tone, and bone health (Walker, et
al.,1997).
In men, 95% of testosterone is
created (produced) in the testes and the rest
(is produced) (delete) in the adrenal glands.
There are two major forms of testosterone in
human body: bound testosterone and free
testosterone. Bound testosterone is attached
to another substance such as globulin or
albumin. When the testosterone is attached
to another molecule, it is modified and
subsequently cannot fit into a receptor site.
Since it cannot become active, it is
eventually excreted by the body. Free
testosterone is the amount of testosterone
that floats through (circulates in) the blood
on its own and is readily available for use by
the body. In men, 35 to 155 pg/mL
(picograms per milliliter) is the normal
range of free testosterone. There may be
daily variations (in the concentration of
testosterone in the body).
as male (delete: as male) testosterone
levels peak in the morning and then steadily
decline until late evening. Stress, lack of
sleep and many other factors can affect
testosterone levels (Meuser, 1977).
When getting (measuring) the
testosterone levels tested (delete: tested), the
results may be misleading. If the level of
bound testosterone is within normal range, it
is not necessarily an indication of healthy
levels. The level of free testosterone is more
important, because free testosterone is the
one that is more readily used by the body
(Dabbs, Mallinger, 1999).
Taking steroid hormones such as
testosterone for performance enhancement
in males can cause testicles to shrink and
breast tissue to growth. For women, it can
cause a deepened voice, an enlarged clitoris,
hair loss from the head, and hair growth on
the body and face. In both genders, steroid
abuse can cause acne, mood swings,
aggression, and other problems. A simple
error in diet, such as (eating a lot of sugar
or)(delete) high-glycemic foods, restricting
fat, being abstaining from meat, or not
getting enough magnesium, zinc, or vitamin
D may lower testosterone levels. These
lowered testosterone levels may affect the
functions of individual development,
immune system, libido, heart health, energy,
weight control and emotional well-being
(Apicella, Feinberg 2009).
In men, the different depths of
voice(voice deepness is) are influenced by
different concentrations of circulating sex
hormones and also by the androgen
sensitivity of the target organs (Meuser,
1977). According to a Georgia State
University experiment, salivary testosterone
levels were “significantly associated with
lower pitched voices among males but not
among females (Dabbs, 1999).” (A similar
study performed by…in)Harries performed a
similar study looking at how testosterone
level affects the males voice during puberty.
Their study (both studies) concluded that
testosterone was not “predictive of the
changes, (but) there was a correlation with
testis volume and testosterone levels."
Alternative test methods, including multiple
saliva samples from the Human Cognitive
Neuroscience Unit of the School of
Psychology and Sport Sciences, indicated
even greater magnitude of the relationship
between high testosterone and lower voice
frequency. Some (studies, researchers….?)
have hypothesized that lower voices have
had an evolutionary benefit of attracting
females by demonstrating a high level of
testosterone. (This has been debunked,
however,)(delete) as a A Harvard study
demonstrated that while men prefer higher
pitched women, women express no
preference for the reverse on males
(Apicella, 2009).
(Our study tests)(delete) In this study
the correlation between (the)(delete)
testosterone levels and the depth of voice in
men was tested. Since the testosterone
fluctuates considerably from one time to the
next we (use third person) decided to collect
multiple saliva samples and use the average
of testosterone levels for the calculations.
Based on past results, there is testosterone in
saliva, and the average levels detected were
295 ± 36 pg/ml in adult males (Dabbs,
Mallinger, 1999).
(Our)(delete) Researchers
hypothesize that the mean testosterone level
is going to be higher in men with lower
voice frequency.
Materials and Methods (centered)
Ten male adult subjects were used in
this study. Subjects were (adult male)
(delete) students at Saddleback College,
Mission Viejo, California. All
measurements were made on the 20th of
November 2013. Subjects were contacted
before and asked to participate in the study.
Each participant signed an experiment
specific agreement waiver and filled out a
short questionnaire. After agreement was
reached (signed), we (third person)
scheduled the day we (third person) would
collect the saliva and voice samples from the
subjects.
On the scheduled day (November 20,
2013)=no repetition necessary. the subjects
were all present for The first set of samples
were collected collecting between 8:20 to
8:40 am. Each subject was given a saliva
collection aid and a pre-labeled collection
vial. Subjects were asked to insert the
ribbed-end of the saliva collection aid in the
pre-labeled collection vial and allow saliva
to pool in mouth before forcing the saliva
through the saliva collection aid into the vial
and fill approximately half the vial (1 mL)
(awkward sentence). (Then)(delete). The
saliva collection aid was removed and
discarded and the cap was attached to the
collection vial…. (join sentences)… It was
made sure that the subjects passively
drooled into their labeled collection
container. Directly after saliva retrieval
coma the researchers placed the test
containers into a cold storage filled with ice
to freeze the samples preventing
degradation. All the voice samplecollecting occurred outside room SM 244 at
Saddleback College, Mission Viejo,
California. Subjects were asked to make the
lowest note they were able to produce for 3
seconds. This recording was accomplished
by was recorded using an iPhone 4S which
yielded a collection rate of 44coma600
samples per second. The sample-collecting
occurred when no other individual (except
the subject himself and the researcher that
was holding the recording device) was in
approximately 5 yards of the subject to
minimize the background noises in the
recordings as much as possible. The second
set of samples was collected under identical
circumstances between 11:20 and 11:40 am.
The second set of saliva samples was also
placed into a cold storage filled with ice to
freeze the samples preventing degradation.
During the sample-collecting, each sample
was labeled as a number in order to keep the
subject's privacy. In between collection and
analysis the samples were stored in a freezer
at -4 oC. degrees Celsius.
The analysis of testosterone samples
analysis occurred on November 22, 2013 in
room SM 244 at Saddleback College,
Mission Viejo, California. The researchers
brought out all of the sample containers
from the freezer at 8 am so for the samples
would be thawed in time for analysis to be
analyzed. The reagents were also removed
from the refrigerator the day before in order
to come to room temperature. The
Salimetrics preparation and following
appropriate analysis procedure was followed
providing us with our raw data. gave the
initial data.
Voice recordings were analyzed
using Speech Analysis Software (SIL
International, Dallas, Texas) to find the
fundamental frequency (average pitch of the
sample) of the subject's voice. The equation
used by the Speech Analysis Software is a
modified version of Cepstrum analysis
𝐹𝑓𝑢𝑛𝑑𝑎𝑚𝑒𝑛𝑡𝑎𝑙 = 𝐶(𝑥) =
|ℱ −1 {log⁡(|ℱ{𝑓(𝑡)}|2 }|2 where represents
a fourier transformation of the equation.
The results of this equation changed the
wave form into a more linear function
making a calculation of average frequency
possible within the Speech Analysis
Software.
Results (has to be centered)
The data was run through a
Microsoft Office Excel spreadsheet. The
program took the input data obtained from
the microreader and outputted a 4Parameter Sigmoid Minus Curve. The curve
was adjusted to fit along the plot points of
the standards as best as possible (minimum
asymptote: 0.1, steepness: -1.2, inflection
point: 100, and maximum asymptote: 1) (be
consistent with the format). This curve
allows the calculation of the testosterone
concentration in the unknown saliva samples
as demonstrated in Figure 1.
Ratio of optical density (B/Bo)
1.2
1
0.8
0.6
∑𝑁
𝑖=1 𝑡𝑖
2𝑁
= 𝑀𝑒𝑎𝑛⁡𝑇𝑒𝑠𝑡𝑜𝑠𝑡𝑒𝑟𝑜𝑛𝑒
The mean value of the voice frequency level
from all subjects (n=10) was calculated 87.2
Hz.
The subjects were separated into two
groups: the first group includes included the
subjects with lower voice frequency than the
total mean. The second group includes
included the subjects with the higher voice
frequency than the total mean. For each
group, the mean of testosterone level and
voice frequency was were obtained and
analyzed separately.
For the first group (subjects with lower
voice frequency than the total average) the
mean testosterone level was 626.139 pg/mL
(S.E.M ±134.728 pg/mL, C.L. ±428.766).
For the second group (subjects with higher
voice frequency than the total average) the
mean testosterone level was 475.896 pg/mL
(S.E.M ±35.546 pg/mL, C.L. ±91.375).
These data are shown in Figure Two 2.
0.2
0
1
10
100
1000
Log of Concentration (pg/ml)
Figure 1. Graph showing the relationship relating
between the Percent Bound of Standards and
Unknowns with the Log of Testosterone
Concentration (n=10). As the log of concentration
increases, the percent bound for standards and
unknowns decreases. The minimum asymptote
was 0.1, the steepness was -1.2, the inflection
point was 100, and the maximum asymptote was
1.
(label
the standards and the unknowns)
(indentation) After the testosterone
concentrations for each well was were
gathered, they were arranged according to
test and subject. The mean value of
testosterone levels for each subject was
calculated using: the formula
Mean of patients' free
testosterone level (pg/mL)
0.4
800
600
400
200
0
Below Total Average Above Total
Average…
Figure 2. Mean Testosterone Concentration level
compared to the Mean Frequency of Voice in each
subject (n=10). The average voice frequency was
measure 87.2 Hz. There was no significant difference
in the mean testosterone level of subjects with voice
frequency of below total average in compared to the
subjects with higher voice frequency of the total
average. (p=0.179, one-tailed unpaired t-test). Error
Bars are mean ± SEM. (review figure captures for
both figures and make them more concise)
Based on the p value (P=0.179), we (use
third person) can reject our (use third
person) hypothesis and accept the null
hypothesis that there is no significant
difference in the mean testosterone level in
relation to fundamental voice frequency.
Two other studies also concluded that
testosterone levels have no significant
relationship to voice frequency in men
(Hughes et. al, 1997) (Meuser and
Nieschlaq, 1977). In our (use third person)
experiment the testosterone levels in
subjects were significantly higher than the
normal adult males. The possible
explanation for this difference is the
performance errors during the saliva
preparation that could cause the incorrect
ratio of the sample to reagent. This error
could lower the number of reactions
between the antibodies and the testosterone
in Salimetric plates and eventually
demonstrate higher testosterone level than
the actual level.
Discussion….
Acknowledgments…
Citations
Apicella, Coren and David Feinberg. 2009.
(Year goes towards the end) Voice Pitch
Alters Mate-Choice-Relevant Perception in
Hunter-Gatherers title has to be italicized .
Proceeding of the Royal Society. Vol.
276:1077-1082.
Dabbs, James and Alison Mallinger. 1999.
(Year goes towards the end) High
Testosterone Levels Predict Low Voice
Pitch Among Men title has to be italicized
Science Direct. Vol. 27, Issue 4.
Evans, Sarah, Nick Neave, Delia Wakelin,
and Colin Hamilton. 2008. (Year goes
towards the end) The Relationship
Between Testosterone and Vocal
Frequencies in Human Males. title has to be
italicized Science Direct. Vol. 93, Issue 45.
Hughes, J M Walker, D M Williams, and S
Hawkins. 1997. (Year goes towards the
end) Changes in the Male Voice at Puberty.
title has to be italicized Archives of
Disease in Children. 77:445-447.
Meuser W. Nieschlaq. 1977. (Year goes
towards the end) Sex hormones and depth
of voice in the male title has to be italicized
(Article in Germany). Deutsch Med
Wochenschr. 25:102(8):261-4.
Review Form
Department of Biological Sciences
Saddleback College, Mission Viejo, CA
92692
Author (s): Ben Hoffman, Sid Mirgati, Tim
Turnbaugh
Title: The Mean Testosterone Level is
Higher in Adult Males With Deeper Voice.
Summary
Summarize the paper succinctly and dispassionately. Do
not criticize here, just show that you understood the paper.
Testosterone is a steroid hormone
that participates in the development of
genitalia in males and also in the maturation
of puberty. Most of the hormone is produced
in the testis and the rest in the suprarenal
glands. There are two forms of testosterone
in human body: bound and free testosterone.
Free testosterone is the one that is readily to
use for the bound testosterone is attached to
other molecules and eventually excreted. In
this study, the researchers collected saliva of
participant individuals at two times on a
specific day to measured the concentration
of the hormone and used an intelligent
phone to measure the deepness of their
voice. Researchers hypothesized that males
with deeper voice would have higher
concentration of the hormone compared to
average or higher voice frequency. The
result indicated that there is no relation
between the concentration of the hormone
and the deepness of the voice.
General Comments
Generally explain the paper’s strengths and
weaknesses and whether they are serious, or
important to our current state of knowledge.
The strengths
The explanation of the importance of
Testosterone in our body, including its
normal range in males, factors that affect its
concentration in the body, and the effect of
ingesting the hormone gave a good
introduction about this steroid hormone.
Methods were well explained, and the
results were correctly analyzed using the
appropriate statistical analysis.
The weaknesses
Even though, methods in the paper are well
explained, the collection of saliva samples
were not controlled and researchers did not
provide the appropriate conditions or
environment to collect voice frequencies,
because they were measured outside the lab
room, where a lot of secondary noise in the
background could have given false
measurements.
Paper needs correct formatting and better
figure capturing
Needs Discussion.
Overall
The knowledge of testosterone in male
bodies is appropriate to the public to
understand the importance of its function,
especially for men. Knowing this, adequate
diet can be followed to maintain proper
levels in the body; and also, the secondary
effects can be known if consumed
artificially.
A better sample collection and better control
of the environment would have made this
study a good candidate for publication.
Technical Criticism
Review technical issues, organization and clarity.
Provide a table of typographical errors, grammatical
errors, and minor textual problems. It's not the
reviewer's job to copy Edit the paper, mark the
manuscript.
This paper was a final version
This paper was a rough draft (check
mark here)(this is a rough draft)
Observe all the corrections made in
the article. High amount of
grammatical and typographical
errors discovered.
Recommendation
 This paper should be published as is
 This paper should be published with
revision
 This paper should not be published (paper
should not be published)