Ethics in Youth Sport and Exercise Training Assignment Instr.docx

Ethics in Youth Sport and Exercise Training
Assignment Instructions
During the past eight weeks, the class put together and
conducted a research project. As well, each week you have been
preparing your research report on that project. For this
assignment, you need to compile the different parts into one
paper and submit it for your final grade. Several sections have
already been graded, and you have been provided feedback.
You can submit what you originally submitted and rewrite the
sections incorporating the suggestions of your classmates and
the instructor. (Remember, your Turn It In Report will
showcase anything that has been cut and pasted. You need to do
a heavy edit on anything previously submitted.)
You will add one new section, the discussion and conclusions.
For this section, use your review of the literature and annotated
bibliography to justify the results of your findings from the
survey and to critically support what you found.
This is the final product of what you have been working on for
7 weeks.
The format of your paper is:
Maximum page limits
and order
Content
1
Title, Author, and date
1
Abstract
2
Introduction
4
Review of Literature
2

Methods
Unlimited
Results
1-4
Discussion and Conclusions
Unlimited
References
Formatting Guidelines for Research Paper
· Double spaced
· Times New Roman font
· 12 font
· 1" margin all the way around
· Reference list (minimum of five references, APA Format)--
remember your references must be from scholarly and/or
primary sources (the textbook is considered neither).
Week 2
The first article, “Concussion in Sport: Conceptual and Ethical
issues,” McNamee et al. (2015) affirms that sport injuries,
particularly, concussions have prompted ethical debates on their
management. The issue seems to generate divisive opinions on
how concussions are diagnosed and treated in sports which may
in turn harm the well-being of athletes. This article is
instrumental to the topic because it provides a comprehensive
understanding into how professionals and scientists handle the
ethics of managing concussions among athletes while
prioritizing their health and career dreams.
The second article, “ Ethical Issues Surrounding
Concussions and Player Safety in Professional Ice Hockey,”
Caron and Jeffrey (2015), aim to provide effective solutions to
dealing with concussions which endanger their safety as well as
managing well the ethical issues that arise from such
circumstances. This helps to further add knowledge into the

number of cases currently documented as well the outcomes of
interventions. Additionally, the authors are referring to previous
research which helps to establish any correlations between
concussions and the relationships and quality of life of athletes.
The third article, “Moral/Ethical Analysis of
Performance Enhancement in Sports,” McCalla and Neil (2014),
provide an in depth analysis of the moral and ethical view of
performance enhancement drugs in sports. Ethically, the authors
apply the Jeremy Bentham theory and Kant’s Deontology theory
to help analyze the actions of athletes in this issue. The authors
assert that all athletes need to show good faith through showing
honesty and integrity. This article is valuable to the topic
because it broaches a controversial issue in sports which has led
to dearth in the careers of athletes and other officials as well as
putting into doubt the achievements made by sporting
organizations such as the Olympics.
Ethical issues in sports was entered into the APUS Tefry
Library search enginee and peer-viewed was selected as a filter.
References
Caron, Jeffrey, and Bloom, Gordon. (2015). “Ethical Issues
Surrounding Concussions and Player Safety in Professional Ice
Hockey.” Neuroethics 8(1), 5–13. Web.
Mcnamee, Michael, Partridge, Bradley, and Anderson,
Lynley.(2015). Concussion in Sport: Conceptual and Ethical
Issues. Kinesiology Review 4(2), 190–202. Web.
Mccalla, Sandra, and Shepherd, Neil.(2014). MORAL/ETHICAL
ANALYSIS OF PERFORMANCE ENHANCEMENT IN
SPORTS. International Journal of Arts & Sciences 7(4), 371–
381. Web.
Annotated Bibliography
Armstrong, Neil et al. “Aerobic Fitness and Its Relationship to
Sport, Exercise Training and Habitual Physical Activity During
Youth.” British journal of sports medicine 45.11 (2011): 849–
858. Web.
The aim of this study was to analyze aerobic fitness and its

correlate with sports involvement, habitual physical activity,
and exercise training. The study was done through sports
computer search of Medline, sports database and personal
databases. The results indicate that VO(2) increase with
maturity.
Hockey.” Neuroethics 8.1: 5–13. Web.
The authors of this article investigated ethical issues around
concussions in professional sports and players safety among Ice
hockey. The results of the study indicate that taken together,
concussions impact many people in the sporting community
from current and former professional athletes and their families
to medical and health professionals and researchers.
Greenfield, Bruce H, West, Charles Robert, and Greenfield,
Bruce H. (2012). “Ethical Issues in Sports Medicine: a Review
and Justification for Ethical Decision Making and Reasoning.”
Sports health 4.6: 475–479. Web.
The authors of this study conducted a study using systematic
review of literature to investigate the impact of an ethical issue
in sports on healthcare in people working with athletes. The
finding of the study indicates that the multiple stakeholders
involved in sports teams challenge the traditional notion of
confidentiality and autonomy.
Hums, Mary, Barr, Carol, and Gullion, Laurie. (1999). “The
Ethical Issues Confronting Managers in the Sport Industry.”
Journal of Business Ethics 20.1: 51–66. Web.
The authors in this study focused on understanding the ethical
issues that managers in the sports industry face today. The
authors of this study revealed that the sports industry is
experiencing rapid growth and development, and as it grows,
sport managers in the different segments encounter ethical
issues, which are often unique to each segment.
Lewandowska, Aleksandra and Leźnicki, Marcin. (2016).
“Doping w Sporcie Jako Problem Etyczno-Społeczny = Doping
in Sport as a Problem Ethical and Social.” Journal of Education,

Health and Sport 6.7: 548–555. Web.
The article presents the issues of doping in sport used in the
context of possible social consequences associated with its use,
as well as at the level of biomedical related to the rapid
development of biomedical technologies in the area of
human body enhancement.
Mccalla, Sandra, and Shepherd, Neil. (2014). “Moral/Ethical
Analysis of Performance Enhancement in Sports.” International
Journal of Arts & Sciences 7.4: 371–381. Web.
The authors of this study focused their study on the impacts of
performance enhancement and how they influence the ethics in
professional sports. From this ethical perspective, we combine
Jeremy Bentham's Utilitarian theory with Kant's Deontology
theory to show how a combination of both theories can provide
one possible response to the ethical actions of athletes in
respect to the use of performance enhancers. The authors
recommend for the formation of 'self duty' that all athletes
should have to the ethical rules as is applied to honesty and
integrity.
Mcnamee, Michael, Partridge, Bradley, and Anderson, Lynley.
(2015) “Concussion in Sport: Conceptual and Ethical Issues.”
Kinesiology Review 4.2: 190–202. Web.
The authors of this article are of the views that scientists,
academia, legal, and professional medics have not been able
speak with one voice on how to curb concussion in sports. The
authors discuss this issue of concussion from philosophical,
bioethical, and sports ethical perspectives.
Polly, D. (2013). Avoiding Ethical and Legal Issues in Practice
Settings. Journal of the Academy of Nutrition and Dietetics
113.12: n. pag. Web.
The authors of this article are of the views that dietetics
practitioners encounter many ethical and legal gray areas in
professional practice every day. These issues may include
patient-related decisions, appropriate business practices, or
relationships with professionals, customers, and employees. It is
because of this that the authors are of the views that most of the

professionals are facing legal battles in the courts because they
violate the ethics in their practices.
Phillips, S. (2012). Carbohydrate Supplementation and
Prolonged Intermittent High- Intensity Exercise in
Adolescents. Sports Medicine 42.10, 817–828. Web.
The authors of this article are of the idea that there has
research in the last decade to investigate the efficacy of
carbohydrate supplementation for improving aspects of physical
capacity and skill performance during sport-specific exercise in
adolescent team games players. However, they assert that the
research is still at its youngest stage and there is need to
enhance it so that the results can be achieved and used to
implement positive policies in youth sports.
References
858. Web.
Hockey.” Neuroethics 8.1: 5–13. Web.
Hums, Mary, Barr, Carol, and Gullion, Laurie. (1999). “The
Ethical Issues Confronting Managers in the Sport Industry.”
Journal of Business Ethics 20.1: 51–66. Web.
Lewandowska, Aleksandra and Leźnicki, Marcin. (2016).
“Doping w Sporcie Jako Problem Etyczno-Społeczny = Doping
in Sport as a Problem Ethical and Social.” Journal of Education,
Health and Sport 6.7: 548–555. Web.
Mccalla, Sandra, and Shepherd, Neil. (2014). “Moral/Ethical
Analysis of Performance Enhancement in Sports.” International
Journal of Arts & Sciences 7.4: 371–381. Web.

Mcnamee, Michael, Partridge, Bradley, and Anderson, Lynley.
(2015) “Concussion in Sport: Conceptual and Ethical Issues.”
Kinesiology Review 4.2: 190–202. Web.
Polly, D. (2013). Avoiding Ethical and Legal Issues in Practice
Settings. Journal of the Academy of Nutrition and Dietetics
113.12: n. pag. Web.
Phillips, S. (2012). Carbohydrate Supplementation and
Prolonged Intermittent High- Intensity Exercise in
Adolescents. Sports Medicine 42.10, 817–828. Web.
Week 3
Problem statement: there is a link between concussions and the
physical and mental health of children and youth in sports
Hypotheses: a) What is the frequency of concussions among
youth and children who play sports and what are the reporting
rates? b) How are concussions diagnosed and treated and are
they perceived based on the mental and physical health of
athletes? c) What are incidence rates of children and youth who
suffer concussions in regards to serious physical injury and
depressive moods including other mental disorders?
The incidence rates of concussions in sports among children and
youth
Most schools and colleges have rolled out sports
programs which allow children and youth to participate in
varsity games at mostly competitive levels. While some sports
such as swimming are less injurious others like rugby, soccer,
and hockey can be very aggressive leading to many cases of
concussions. This has already caught the attention of parents
who are anxious about the type of sports their children should
participate in, the safest equipment to use, and when to
effectively determine if their child can resume sports after
suffering a concussion (Graham et al., 2014). The reports
indicate that between 2001 and 2009, the rate of concussions
and other non-serious injuries among youth from age 19 years
and below increased from 150,000 to 250,000 (Graham et al.,

2014). The almost 100% increased incidence rate is definitely a
worrying trend for parents, teachers, and coaches.
McNamee et al. (2015) reiterates that concussion in
sports has gained global public interest leading to many
disagreements and conflicts among educational, legal, and
medical professionals and scientists. As a matter of
consequence, looking into the type of sports that increase the
chances of concussions is important. Pfister et al. (2016)
conducted a systematic review and meta-analysis to evaluate the
incidence of concussions among youth athletes in 36 out of 698
studies. The author applied the sources in Medline, Embase and
SportDiscus based on the criteria of study design of prospective
cohort study, relevant sports (rugby, American football and
basketball among others), population, and outcome
(concussion). This study was definitely encompassing based on
the many variables applied and thus helped to reach a definite
and solid conclusion. Pfister et al. (2016) concluded that among
the 12 sports chosen there are significant and sharp differences
in the rates of concussions. The probability is that the authors
settled that sports which tend to have an aggressive and tackling
style of playing such as rugby, American football, and hockey
increases chances of bodily harm including concussions. In
addition, athletic exposure which is a measure of concussion
rates is expected to be slightly higher among male athletes that
female athletes.
The diagnosis and treatment of concussions among children and
youth
McNamee et al. (2015), in their study review the
similarities and differences of leading consensus statements
which provide guidelines for treating of concussion diagnosis
and treatment in sports. This appears to be a preliminary action
taken by the sports and medical fraternity after possibly
witnessing cases on misdiagnosis and poor treatment of athletes
who suffer from concussions. This is also matter of emergency
because children and youth tend to still be developmental stages
and concussions and significantly impact their mental and

physical aptitude. The consensus statements according to
McNamee et al. (2015) target coaches, trainers, physicians,
therapists and other parties which deal with injured athletes. In
particular, the children and youth are given focus under the
pediatric concussion criteria which look into the symptoms,
screening and treatment that is appropriate for their ages to
avoid further harmful consequences (McNamee et al., 2015).
The subject of concussions definitely requires a close study
including of the telltale signs of concussions. McNamee et al.
(2015) points out one of the similarities of the consensus
statements which involves the definition of a concussion. A
concussion in general terms is, “defined as a complex
pathophysiological process affecting the brain, induced by
traumatic biomechanical forces,” (McNamee et al., 2015,
pg.191). It is clearly some form of brain injury which can
seriously undermine the mental and physical development of
children and youth. Greenfield et al. (2012) posits that one of
the obstacles faced in attending to sports-related concussions by
healthcare professionals including team physician, physical
therapist and athletic trainer is ensuing the athlete is in the best
condition to get back to playing. This is clearly a sticky issue
which generates a myriad of ethical and moral issues. When
someone suffers a concussion sometimes they do not show any
external injuries except a few minor cuts and scrapes. If the
child or youth is sent back to play, the physical activity could
easily worsen a hidden brain injury or internal bleeding which
was incorrectly undiagnosed and sometimes leading to death.
This is undoubtedly a big fear for most parents, teachers and
coaches. However, for any keen individual actively engaged in a
child’s or youth’s sports career, it is essential to have basic
knowledge of the signs and symptoms. Graham et al. (2014)
outlines a comprehensive list of such signs and symptoms
including a dazed appearance, confusion about assignment,
forgetfulness, losing consciousness among others. Parents and
teachers also need to pay keen attention to what a child or youth
complains of such as a headache or “pressure” in the head,

nausea, dizziness, light or noise sensitivity or feeling emotional
or nervous among other red flags (Graham et al., 2014).
The impact of concussions on the mental and physical well-
being of children and youth in sports
On a broader platform, Caron and Bloom (2015) reveals
that professional hockey players exposed how concussions have
impacted their professional careers, interpersonal relationships
and qualities of life. This revelation sets precedent for what the
future could be like in the event that the children and youth
would want to pursue sports as a lifelong sports career. This can
act as a form of caution for them to protect themselves from
life-altering physical injuries and mental disorders in the future.
Medically and scientifically, researchers have started to link
receptive concussions with Chronic Traumatic Encephalopathy
(CTE) which is an injury that affects the brain structure
indicated by deposits of tau protein in specific parts of the
brain. In lay man terms, anyone diagnosed with CTE is a
prelude to later having depression, confusion, aggression,
suicidal ideation, and memory loss among others. This grim
outlook of the eventuality of the life of an individual who
repetitively suffers from concussions. Children and youth are
not excluded from this type of degenerative brain disease and
are more at risk if they play sports such as tackle football which
is very violent. It is unfortunate that children as young as 5 are
participating in this type of sport and could possibly manifest
into serious physical and mental problems in adulthood. Caron
and Bloom (2015), concludes that the impacts of concussions
are multilevel affecting athletes, their families, medical, and
health professionals and researchers. It brings into question
whether the parties involved promote fairness and honesty as
the foundation of competitive sports (McCalla & Shepherd,
2014). Ethically, the mentors of children and youth in sports
must always promote sportsmanship while ensuring they are not
harmed in any way. Unfortunately, sometimes the sports played
tend to be very combative and competitive and coaches tend to
push this group too hard when they should also focus on other

areas of their development such as social skills.
References
Caron, J., & Bloom, G. (2015). “Ethical Issues Surrounding
Concussions and Player Safety in Professional Ice Hockey.”
Neuroethics 8(1), 5–13. Web.
Graham,R.,Rivara,F.P., Ford,M.A.,& Spicer,C.M.(2014).
Sports-related concussions in youth : Improving science,
changing the culture. National Academy of Sciences.
McNamee, M., Partridge, B., & Anderson, L. (2015).
Concussion in Sport: Conceptual and Ethical Issues.
Kinesiology Review 4(2), 190–202. Web.
McCalla, S., & Shepherd, N.(2014). MORAL/ETHICAL
ANALYSIS OF PERFORMANCE ENHANCEMENT IN
SPORTS. International Journal of Arts & Sciences 7(4), 371–
381. Web.
Pfister,T, Pfister,K.,Hagel,B.,Ghali,W.A.,&
Ronksley,P.E.(2016). The incidence of concussion in youth
sports: a systematic review and meta-analysis. British Journal in
Sports Medicine, 50(5), 292-297.
The article I chose is, “Moral/Ethical Analysis of Performance
Enhancement in Sports.” I found the article in the APUS
TEFRY Library. I picked this article because it touches on the
very sensitive and ethical issue of performance enhancement in
sports. The authors apply the ethical perspective of utilitarian
theory and deontology theory which provide a strong framework
on how society views what is right or wrong. Interestingly, the
authors also posit the concept of self-duty which can be a form
of breakthrough to ensure that all athletes cease from falling
under the pressure of performance enhancement to win. I
searched the article by inputting keywords, “Ethical issues in
sports” and chose the peer reviewed category. This article

effectively contributes to the research topic because it discusses
the issue of enhancement in sports which a serious ethical
matter within the ethical framework and actually integrates real
life athletes whose careers have been damaged because of the
scandal of performance enhancement. The variables being
explored include regulation of use of performance enhancement
drugs, use of performance enhancement drugs by athletes and
the present ethical frameworks that govern such actions in
sports. The hypothesis being tested by the authors is whether
there is centrally moral question or not or if it is just a medical
or empirical question.
References
McCalla,S., & Shepherd,N.(2014). Moral/Ethical Analysis of
Performance Enhancement in Sports. International Journal of
Arts & Sciences, 7(4), 371-381.Web.
1. The research problem is the link between concussions and
the physical and mental heath in youth and children
participating in sports.
2. Hypotheses
a) What is the frequency of concussions among youth and
children who play sports and what are the reporting rates?
b) How are concussions diagnosed and treated and are they
perceived based on the mental and physical health of athletes?
c) What are incidence rates of children and youth who suffer
concussions in regards to serious physical injury and depressive
moods including other mental disorders?
3. Survey questions
In order to ask the right survey questions and obtain the right
answers it is important to first seek the informed consent of
some youth and children who are active in sports particularly
those keen on seeking sports scholarships to college. This
requires the presence of their parents especially for those under
the age of 18 years. The survey questions entail:
a) How often are you called regarding your child’s injuries and
how often is it treated as a concussion?

b) How long after your child is treated of concussion does he or
she resume to play in sports
c) Have you noticed any strange behavior or physical state in
your child ever since they were diagnosed and treated of
concussion?
Week 4
Week 4 Assignment: Research Paper Introduction
Just like it is in other organized industries, the sports industry
is also regulated with established rules and laws to streamline
sporting activities. The number of youths that participate in
sports today has increased drastically and it is because of this
that there is need to emphasize on the laws, ethics, and rules
that regulate youth sports (Armstrong et al, 2011). One of the
major areas of focus in youth sports is exercise training where
the major issue of concern has been the quality of training that
is provided to the youths who are involved in sports. Several
studies that have been done on this issue reveal that most of the
youth sports programs do not have the required resources to
employ professional trainers who understand the rules and
regulations that are required to provide not only quality training
to the athletes but in an ethical manner (Mccalla & Shepherd,
2014).
Youth sports programs rely on volunteers as trainers and as
coaches who may not have the required experience to manage
the training and the program as a whole. The volunteer coaches
and trainers have also been reported to have been involved in
belligerent and violent acts that violate the morals required to
be adhered to in managing the youth sports events. Volunteer
coaches and trainers are faced with different legal and ethical
issues, which make them violate some of the regulations
involved in training and put the youths in the sports at risk
(Wiersma et al, 2005). The main problem is that the volunteer
coaches and trainers are not trained on the link between
concussions and the physical and mental health in youth and

children participating in sports. Lack of this knowledge among
the trainers has put youths in the sports program at risk and
threatened their health and wellbeing. This study will focus on
identifying some of the ethical issues that are involved in youth
sports training and exercising.
References
858. Web.
Mccalla, Sandra, and Shepherd, Neil. “Moral/Ethical Analysis
of Performance Enhancement in Sports.” International Journal
of Arts & Sciences 7.4 (2014): 371–381. Web.
Wiersma, Lenny D, Sherman, Clay P, and Wiersma, Lenny D.
“Volunteer Youth Sports Coaches’ Perspectives of Coaching
Education/certification and Parental Codes of Conduct.”
Research quarterly for exercise and sport 76.3 (2005): 324–338.
Web.
Hook example one
Just like it is in other organized industries, the sports industry
is also regulated with established rules and laws to streamline
sporting activities. The number of youths that participate in
sports today has increased drastically and this is the reason
emphases should be put on the laws and regulations that
minimize risks involved in this sports (Armstrong et al, 2011).
Hook example two
The number of youth sports programs has increased drastically
in the recent past (Mccalla & Shepherd, 2014). However, youth
sports programs do not have the required funds to employee
professional coaches and trainers which put the youth involved
at risks of getting injuries during training and exercise.
In both examples, the first sentence raises curiosity in the
readers since they leave the readers in a state of dilemma, which
will want them to read on and understand issue that is raised by
the first sentence. The second sentence in each case grabs the

attention of the readers, as most of the readers may be parents
whose children are involved in sports programs and will want to
read on and understand the risks that are associated with the
programs. The two hooks connect to the research as the readers
and the research alike will want to explore the study and
identify the risks involved in youth sports and how they can be
mitigated.
References
858. Web.
Mccalla, Sandra, and Shepherd, Neil. “Moral/Ethical Analysis
of Performance Enhancement in Sports.” International Journal
of Arts & Sciences 7.4 (2014): 371–381. Web.
Wiersma, Lenny D, Sherman, Clay P, and Wiersma, Lenny D.
“Volunteer Youth Sports Coaches’ Perspectives of Coaching
Education/certification and Parental Codes of Conduct.”
Research quarterly for exercise and sport 76.3 (2005): 324–338.
Web.
The author of this study is of the views that proper aggression
and anger measuring scale must exist and should be grounded in
valid and reality to measure anger and aggression in sports. This
study is a critique of the current Competitive Aggressiveness
and Anger Scale that was developed by Maxwell and Moores in
2007. The critique was done through a review of literature that
focused on three areas of the Scale such as the definition of
aggression that was used in formulating the scale, the one size
fits all approach of thinking that was adopted by the developers
of the scale and the general nature of Competitive
Aggressiveness and Anger Scale relating to anger and
aggressiveness. The author is of the views that the approach that
was used to develop the current CAAS limits the ability of the
scale to measure aggressiveness and anger in sports.

The author concludes that the scale ignores legal and
sanctioned aggressive and violent behaviors in sports, which
limit its ability to understand aggressiveness and anger in
sports. The author divided the study into three sections with
each section addressing a single issue that the author was
investigating. This was important as it allowed the author
thoroughly investigate each element and create a formidable
conclusion based on the available literature. Concentrating on a
single aspect under investigation in each section allowed the
author to effectively deal with the hypothesis. From my own
evaluation of the study, I am of the views that the author
through his reliance on literature and analysis of data, the
author’s conclusion seems to be valid since the conclusion is
founded rather supported by strong evidence derived from the
analysis of data and literature.
Reference
Kerr, John H. “A Critique of the Development of the
Competitive Aggressiveness and Anger Scale.” Psychology of
Sport & Exercise 9.6 (2008): 721–728. Web.
Week 5
Survey QuestionS
Top of Form
Question Title
* 1. How often are you called regarding your child’s injuries
and how often is it treated as a concussion?
w 0
1
2
3
Question Title
* 2. How long after your child is treated of concussion does he

or she resume playing in sports? w 0
1 week
24 hours
Upon Physician or Medical Clearance
Question Title
* 3. Have you noticed any strange behavior or physical state in
your child ever since they were diagnosed and treated with a
concussion w 0
Yes
No
Question Title
* 4. Are the coaches required to take a concussion education
and training course? w 0
Yes
No
Selected Article: “Running with a Mask? The Effect of Air
Pollution on Marathon Runners’ Performance from the Journal
of Sports Economics. Found at:
https://doi.org/10.1177/1527002518822701
Description of Study and Results
Air pollution affects the performance of athletes. It has been
determined by existing literature that air pollution has negative
effects on short-run labor productivity. To add to this body of
literature, Guo and Fu (2017) used a sample of more than
300,000 runners in 55 different races and 37 cities in China
during the period 2014-2015 to estimate the causal impact of air
pollution on the performance of marathon runners. The focus of
the performance evaluation was on the finish time of the
runners used in the study. Guo and Fu (2017) estimated the
elasticity of air pollution of finish time to be somewhere around
0.0408 seconds. The impact is economically important
following the large differences in the quality of air across
different cities sampled in China.
The researchers gathered data and information for the

mentioned races and marathon runners from the site-
from www.runchina.org.cn. The site is under the management
and maintenance of the Chinese Athletic Association. The site
was resourceful for the study because of its publication of finish
time data for athletes for all half-marathon and full-marathon
races that have been held in the country over the last five years.
The data collected for the study was from 2014 to 2015. The
researchers then downloaded the daily air quality index- AQI
information from the Ministry of Environmental Protection of
China at the city level. Guo and Fu (2017) identified the causal
effect of the quality of air on the finish time of a marathon
runner by relying majorly on the air quality heterogeneity on
the day of racing.
The study made some significant results. Firstly, it determined
that an increase in the air quality index results in a significant
increase in the finish time of the marathon runners. This finding
implies that when the quality of air is reduced or minimized, the
average time taken by the marathoners to finish their races is
increased. For instance, in its cross-sectional results, a one-unit
increase in the quality of air index results into the finish time
increasing by 2.7 seconds. The overall finding both for the half
and full marathon runners is uniform and consistent across all
the age groups used in the study- air pollution negatively affects
the performance of the runner.
Conclusion of the Study
Guo and Fu (2017) conclude that air pollution has a significant,
negative impact on the performance of the runners. The causal
identification used in the study which is based on the
heterogeneity of the quality of air on the race day of the race
helps to make the process random. Using the panel dataset of
runners who participated in more than a single race, the
estimates used in the study confirm that air pollution negatively
affects the performance of runners.
Evaluation and Personal Opinion
This study significantly and positively contributes not only
towards understanding the effect of air pollution on exercises

and sports, but also short-term productivity. The findings listed
and inferences drawn from them remind governments and
municipalities that the negative impact of air pollution on
performance and the general health of sportsmen and women
should be greatly considered when organizing for large scale
outdoor sports activities like Olympics. The findings can also
inform professional runners or athletes competing in large scale
events like soccer, biking and running. The findings can also of
great interest and help to employees who work under intensive
physical exercise and are exposed to long periods of ambient air
pollution.
From a personal standpoint, ambient air pollution affects many
aspects of human life- sports and exercises being one of them.
The article brings forth an important aspect of organizing mega
sports events- that organizers must evaluate the impact of air
pollution on the runners. It is true that air pollution has
unfavorable effects on the cardiovascular and respiratory
systems of humans and therefore exposing athletes to such
pollution is dangerous not only to their performance on the
track but also general health. For the athletes, the toxicity of the
pollutants can damage their respiratory and vascular systems by
inflaming the airways.
References
Guo, M., & Fu, S. (2017). Running with a Mask? The Effect of
Air Pollution on Marathon Runners’ Performance. Journal of
Sports Economics, 1527002518822701.
Types of Study Design
Cited Source: Lee, S. Y. (2018). Analysis of relationship
marketing factors for sports centers with mixed methods
research. Asia Pacific Journal of Marketing and
Logistics, 30(1), 182-197.
The kind of study: This study is a mixed research design. As
supported by existing evidence, the mixed methods approach is
where the researcher collects, analyzes and interprets both

quantitative research- which involves surveys and experiments
and qualitative research which may involve interviews and
focus groups.
Why is it this type of research? In this research study, Lee
(2018) uses both a statistical survey, which is a form of
quantitative research and in-depth interviews which is a form of
qualitative research method in studying the impact of
relationship marketing factors of sports centers on commitment
to relationship and recommendation intentions. In its findings,
the quantitative aspect of the research established that the
relationship marketing factors like price, bonding and facility
affect commitment to relationship in a positive manner.
References
Lee, S. Y. (2018). Analysis of relationship marketing factors for
sports centers with mixed methods research. Asia Pacific
Journal of Marketing and Logistics, 30(1), 182-197.
Week 6
An example of inductive reasoning is like saying; Tim is a
rugby player. All the other rugby players within the school team
weight more than 180 pounds. Therefore, Tim must weigh more
than 180 pounds. This is a good example of an inductive
reasoning statement. An example of deductive reasoning is like
saying; Cacti are vegetation, and all forms of plants go through
photosynthesis; therefore, cacti take part in photosynthesis.
Response
Thank you for your well-researched information. I conquer that
one example of inductive reasoning is when one says; Tim is a
rugby player. All the other rugby players within the school team
weight more than 180 pounds. Therefore, Tim must weigh more
than 180 pounds. This is a good example of an inductive
reasoning statement (Johnson-Laird, 1999). An example of
deductive reasoning is like saying; Cacti are vegetation, and all
forms of plants go through photosynthesis; therefore, cacti take

part in photosynthesis. Thank you for your well-researched
information.
References
Johnson-Laird, P. N. (1999). Deductive reasoning. Annual
review of psychology, 50(1), 109-135.
The article to be referenced in this part looks at the
development of types of foods used for nutrition. The research
looks at foods that come from fish products and which are
utilized to build the body for athletes. The research was trying
to look at the best foods that would be ideal for muscle
building. The objects of study that were used were minced cold
fillet that was prepared as per the best standards that are
supposed to be met. Several stages were gone through in trying
to look at the nature of the minced cold fillet that included
planning, examination, and determination. All these stages were
put out in order to find out the components in meat that are
ideal when it comes to practicing athletes (Kolman et al., 2018).
The experiment revealed that food substances are ideal as they
help athletes build muscle that is crucial for them when it comes
to performing their daily tasks.
Looking at the article, it is no doubt that it was in tandem
with the outcome that was needed to look at the best conditions
that are required by athletes. Athletes require a healthy schedule
that would help them maintain a healthy lifestyle that would be
ideal for their muscles and their body functioning. The study
was undertaken by looking at essential variables needed by
athletes, and that shows what is essential when it comes to
looking for the best diet that is required by athletes. There is
with no doubt that this research was good by looking at the
purpose that it was supposed to serve of, finding the right
components in food (Kolman et al., 2018). It goes without
saying that athletes have suffered for a long time as they do not
know the right protein components that they should take.
However, with this research, a majority of athletes will be
informed on which products would suit them.

References
Kolman, O., Ivanova, G., Kudryavtsev, M., Gavrilyuk, O.,
Osipov, A., & Ivanova, A. (2018). Development of new
combined sports nutrition products. Journal of Physical
Education and Sport, 18, 401-407.
Week 7
CONCUSSIONS IN YOUTH SPORTS
This research paper consists of the various procedures the class
used to gather and interpret relevant information concerning the
frequent occurrence of concussions in the youth sports sector.
The article brings about the different methods and study
strategies applied by our research group to collect data during
the procedure. The participants of the research and equipment
used during the experimentation are also evident in the paper.
The results section is apparent later in the article, where the
core findings and interpretations concerning the research
methods are available.
A concussion, as described by McCrory,et.al (2017), is a
traumatic brain injury that takes place after an impact to the
head, and may at some point lead to a state of unconsciousness.
This condition is mainly concurrent during a car accident, fall,
recreational, or sporting activities. Even though concussions are
usually not considered life-threatening, they can still impose
severe symptoms that require medical attention. That is why the
class decided to research the frequent occurrence of
concussions, especially in youth sports. This is because it has
been estimated that a significant fraction of annual concussion
cases happens during sporting and recreational activities, and
majorly among children and youths aged between 14 to 19
years, (McCrory,et.al, 2017).
The research was conducted between two different learning
institutions. The participants for the study were 20 students, ten

from each institution, their parents and guardians, and trainers
from the sporting activities the students participated in. Our
crew selected four sports: football; hockey; basketball; rugby,
from which we chose both the coaches and their assistants to
interview on the players’ injury rates. The group also went
ahead and interrogated the secretaries who were working for the
designated learning institutions. In total, the study comprised of
58 participants:
1. Twenty students- The trainees who participated in the four
sports
1. Twenty parents- The students’ guardians
1. Sixteen couches- The trainers who coached the students
during their games, and their assistants
1. Two secretaries- The employees who worked for the two
institutions
Apart from the participants, the group also used a Mixed-
Method kind of study to collect and interpret the relevant
information needed to generate the findings. As stated by
Östlund, Kidd, Wengström & Rowa-Dewar (2011), mixed-
research studies comprise of both quantitative design method,
which majorly uses statistical survey and qualitative technique,
which often consist of individual interviews and observations.
Shortly before initiating the survey, the research crew selected
two elementary schools to work on, and randomly picked ten
students from each school. Our group chose the four sports
since they were considered the games with the highest incident
in concussion cases, (Majerske, Mihalik, Ren, Collins, Reddy,
Lovell & Wagner, 2008). During the qualitative research
method, the study used individual interviews with all the
participants. The experiment began by interviewing the 20
students, one by on. The selected examiners narrated a story of
previous concussion occurrences to the student, the signs and
symptoms, and finally asked questions of how the student’s
injury occurred, the effects and how long after treatment did the
student resume his or her sporting activity. Later, after all the
students were interviewed and useful data collected from them,

the examiners turned to the parents and guardians. Interview
procedure was similar to that of the students, except the
questioning time, where the interviewers asked questions like
how often the parents were called regarding their offspring’s
injuries and if they had noticed any behavioral or physical
defect in their children since their diagnosis and treatment of
concussion. The couches, on the other hand, were asked
questions concerning their concussion education and training
experiences. Voice recorders were used all through the
interview process to enable future codding of the information.
The study also included observing and participating with the
students during their sporting activities. Our research team went
ahead and applied the quantitative method by accessing the
students’ health records from the two secretaries working for
the schools. The files provided relevant statistical information.
The findings under the results section show that approximately
half of all the concussions went unnoticed. It could have been
because the students’ couches were not aware of their
concussions. 40% of participants students were found to be
playing with signs and symptoms related to concussions due to
lack of attention from their parents and trainers. Regarding the
“how often are you called concerning your child’s injury”
question to the guardian, the results had it that 15% of all the
twenty students reported two or more concussions annually. It
was because a significant number of the students played without
the right protective attire. Additionally, the health records
showed that 9% of all affected students reported the matter but
did not access proper medical attention.
It is, therefore, appropriate to immediately report incidences of
concussion, put on suitable protective clothing, and always
practice a safe sports culture during gaming.
Reference
Majerske, C. W., Mihalik, J. P., Ren, D., Collins, M. W.,
Reddy, C. C., Lovell, M. R., & Wagner, A. K. (2008).
Concussion in sports: postconcussive activity levels, symptoms,

and neurocognitive performance. Journal of athletic
training, 43(3), 265-274.
McCrory, P., Feddermann-Demont, N., Dvořák, J., Cassidy, J.
D., McIntosh, A., Vos, P. E., ... & Tarnutzer, A. A. (2017).
What is the definition of sports-related concussion: a systematic
review. Br J Sports Med, 51(11), 877-887.
Östlund, U., Kidd, L., Wengström, Y., & Rowa-Dewar, N.
(2011). Combining qualitative and quantitative research within
mixed method research designs: a methodological
review. International journal of nursing studies, 48(3), 369-383.
The article: The Role of Social Context in Shaping Student-
Athlete Opinions
Reference: Druckman,J.N.,Gilli,M.,Klar,S. Robison,J.().The
Role of Social Context in Shaping Student-Athlete Opinions.
PLoS One, 9(12).
The study focuses on surveying on the opinions of post-
career student athletes regarding payments. The authors apply a
2012 survey which focuses on the NCAA Big Conference based
in Midwest Nebraska. The survey includes 12 major universities
who participate in the Division I NCAA Athletics. The
limitation of the study was focus on one sample in form of the
Big Conference; however, it provides a significant variance
among universities. The researchers posed two questions to the
respondents one of which was about a proposal to pay athletes.
The second question was about support for unionization of
college athletics. Other variables which the researchers
considered were race, gender, family income, scholarship status,
partisan membership, sport played and year in school. The
result findings indicated that 49.49% of the respondents support
pay for play while 22.02% supported unionization of college
athletics.
Researchers concluded that the post-career changes in
opinions by student-athletes are changed by their social
environment which entails individuals from their university and
team. Overall, the researchers assert that future studies need to

integrate the role of the social environment in influencing
opinions and in this case regarding paying college-athletes.
The article has achieved its purpose which is to seek out
the opinions of student-athletes regarding payment for their
participation in sport. The authors have achieved this by
choosing an appropriate sample which offers all the variables to
help test the hypothesis. In this case, it chose from the Athletics
websites and were able to send at least 5,896 emails. They were
able to obtain 1,303 responses which is 31.8% which is above
most email surveys. The variables which the researchers applied
and the two leading questions help to produce the research
findings they were looking to discover. In this case a
considerable number of students out of the sample obtained
supported payment for athletics performance and unionization
of college athletics. This clearly provides a guide into the
debate of paying college athletes. In this case, it is the athletes
who largely participate in the sports that show a direction that
NACCA and other bodies should adopt.
My overall opinion is that the researchers took a useful
angle I regards to the question of whether college athletes
should be paid. In this case, they conducted a survey of the
actual student-athletes and sought their opinions on the matter.
The response rate was important enough to confirm that college
sports organizers and sponsor should really consider
compensating the athletes. In addition, the researchers
employed other significant variables such as race, gender and
sports played to ensure the research results and conclusion were
not biased.
References
Druckman,J.N.,Gilli,M.,Klar,S. Robison,J.().The Role of Social
Context in Shaping Student-Athlete Opinions. PLoS One, 9(12).
Week 8

Forum 8A
One of the unethical researches is the case of Johnson and
Johnson who violated the FDA rules by producing and
marketing Risperdal. The company irresponsibly prescribed the
drug to treat all kinds of illness in children and the elderly. It
especially marketed to the elderly even thought it was aware
that Risperdal generated serious health risks such as strokes.
The company also had conducted an internal research and was
able to establish that a boy developed large breast after taking
Risperdal (Saunders, 2015). The entire conduct was unethical
because, the organization first lacked transparency on the
research and production of the drug in their labs. Despite
knowing the negative impacts on health the company still
marketed and distributed the drugs. Secondly, the company
violated the FDA directives in terms of marketing and selling
the drugs. It is clear that the company was after making profits
and disregarding the well-being of its clientele.
References
Saunders,J.(2015). Johnson & Johnson unethical and illegal
drug and device marketing: A review of the seven brill article.
Retrieved from https://www.saunderslawyers.cm/jhnson-
johnson-unetical-and-illegal-drug-and-device-marketing-a-
review-of-the-steven-brill-article/
Bottom of Form

Name: Date:
Critical Reading Form – Bundo, et al. 2014
Purpose:In 1-2 sentences, what was the research question the
authors were trying to answer and why?
Central Thesis:Using only 1 sentence, state the authors’ thesis.
What hypothesis were they testing?
Key Claims:What are two statements given by the authors to
support the thesis? (The claims are usually stated in the
discussion or conclusions section of the paper and explain how
the data support or refute their initial hypothesis, ie why it is
important..
1.
2.
Evidence (ie. Experimental Results): What evidence does the
author use to support each of the key claims you identified?
1.

2.
Figure Analysis: Examine Figure 1C and read the results section
of the paper for these experimental data carefully. Answer
questions 1-4 below.
1A. What is shown in the left panel?
B. Which group(s) showed a statistically significant difference
in mean L1 copy number as compared with controls?
2A. What is shown in the right panel?
B. Which group(s) showed a statistically significant difference
in mean L1 copy number as compared with controls?
3A. What kind of genes are HERVH and SATA?
B. Why did the authors normalize the L1 copy number by

comparison to both HERVH and SATA?
4. What conclusion did the authors assert based on this data?
Neuron
Report
Increased L1 Retrotransposition
in the Neuronal Genome in Schizophrenia
Miki Bundo,1,2 Manabu Toyoshima,3 Yohei Okada,4 Wado
Akamatsu,4 Junko Ueda,2 Taeko Nemoto-Miyauchi,2
Fumiko Sunaga,1 Michihiro Toritsuka,5 Daisuke Ikawa,5
Akiyoshi Kakita,6 Motoichiro Kato,7 Kiyoto Kasai,8
Toshifumi Kishimoto,5 Hiroyuki Nawa,9 Hideyuki Okano,4
Takeo Yoshikawa,3 Tadafumi Kato,2,* and Kazuya
Iwamoto1,10,*
1Department of Molecular Psychiatry, Graduate School of
Medicine, The University of Tokyo, Tokyo 113-8655, Japan
2Laboratory for Molecular Dynamics of Mental Disorders
3Laboratory for Molecular Psychiatry
RIKEN Brain Science Institute, Saitama 351-0198, Japan
4Department of Physiology, Keio University School of
Medicine, Tokyo 160-8582, Japan
5Department of Psychiatry, Nara Medical University, Nara 634-
8521, Japan
6Department of Pathology, Brain Research Institute, Niigata

University, Niigata 951-8585, Japan
7Department of Neuropsychiatry, Keio University School of
Medicine, Tokyo 160-8582, Japan
8Department of Neuropsychiatry, Graduate School of Medicine,
The University of Tokyo, Tokyo 113-8655, Japan
9Department of Molecular Neurobiology, Brain Research
Institute, Niigata University, Niigata 951-8585, Japan
10PRESTO, Japan Science and Technology Agency, Saitama
332-0012, Japan
*Correspondence: [email protected] (T.K.), [email protected]
(K.I.)
http://dx.doi.org/10.1016/j.neuron.2013.10.053
SUMMARY
Recent studies indicate that long interspersed
nuclear element-1 (L1) are mobilized in the genome
of human neural progenitor cells and enhanced in
Rett syndrome and ataxia telangiectasia. However,
whether aberrant L1 retrotransposition occurs in
mental disorders is unknown. Here, we report high
L1 copy number in schizophrenia. Increased L1
was demonstrated in neurons from prefrontal cortex
of patients and in induced pluripotent stem (iPS)
cell-derived neurons containing 22q11 deletions.
Whole-genome sequencing revealed brain-specific
L1 insertion in patients localized preferentially to
synapse- and schizophrenia-related genes. To study
the mechanism of L1 transposition, we examined
perinatal environmental risk factors for schizo-
phrenia in animal models and observed an increased
L1 copy number after immune activation by poly-I:C
or epidermal growth factor. These findings suggest
that hyperactive retrotransposition of L1 in neurons
triggered by environmental and/or genetic risk fac-
tors may contribute to the susceptibility and patho-

physiology of schizophrenia.
INTRODUCTION
Mental disorders including schizophrenia, bipolar disorder, and
major depression affect a large proportion of the global popula-
tion and have a major negative economic impact. Twin, family,
and adoption studies indicate the complex involvement of both
genetic and environmental factors for these diseases (Keshavan
et al., 2011). Despite their apparent heritability, however,
causa-
306 Neuron 81, 306–313, January 22, 2014 ª2014 Elsevier Inc.
tive genetic factors are mostly unknown except for rare cases
of schizophrenia associated with chromosomal abnormalities
(Brandon and Sawa, 2011; Cook andScherer, 2008; Karayiorgou
et al., 2010). On the other hand, environmental risk factors
including prenatal infection (Brown, 2006) and obstetric
compli-
cations, such as neonatal hypoxia, embryonic ischemia, and
gestational toxicosis (Lewis and Murray, 1987), are well-estab-
lished risk factors for schizophrenia. However, it is not clarified
how these environmental risk factors interact with genomic

factors.
Accumulating evidence indicates that genomic DNA in the
brain contains distinctive somatic genetic variations compared
with nonbrain tissues (Poduri et al., 2013). These genetic signa-
tures include brain-specific somatic mutations (Poduri et al.,
2013), chromosomal aneuploidy (Rehen et al., 2005; Yurov
et al., 2007), chromosomal microdeletion (Shibata et al., 2012),
and the genome dynamics of nonlong terminal repeat (LTR) ret-
rotransposons (Baillie et al., 2011; Evrony et al., 2012;Muotri
and
Gage, 2006). These observed somatic variations are hypothe-
sized to contribute to the generation of functionally diversified
brain cells (Muotri and Gage, 2006).
Among the known retrotransposons, only long interspersed
nucleotide element-1 (L1) has autonomous retrotransposition
activity. Full-length L1 elements include a 50 UTR, two open
reading frames (ORFs), and a 30 UTR (Figure 1A). Encoded
prod-
ucts from the ORFs contain activities required for
retrotransposi-
tion and are employed in the insertion of new L1 copies as well

as
nonautonomous retrotransposons such as Alu and SVA (Cor-
daux and Batzer, 2009). Recent studies indicate that engineered
L1 has retrotransposition activity in neural progenitor cells
from
rat hippocampus (Muotri et al., 2005), human fetal brain
(Coufal
et al., 2009), and human embryonic stem cells (Coufal et al.,
2009). These in vitro findings were confirmed in human L1
trans-
genic mice in vivo (Muotri et al., 2005). Adult human brain
cells
also showed increased L1 copy number compared with non-
brain tissues (Coufal et al., 2009). Moreover, retrotransposition
mailto:[email protected]
mailto:[email protected]
http://crossmark.crossref.org/dialog/?doi=10.1016/j.neuron.201
3.10.053&domain=pdf
about 6 kb
ORF1
40 kDa

ORF2
150 kDa
3’ -UTR
RNA binding
protein
Reverse Transcriptase/
Endonuclease
5’ -UTR
m5UTR mORF1 mORF2mouse
h5UTR#2
hORF1#5
hORF2#1
hORF2#2
hORF1#1
hORF1#3human
macaque cORF2#4
cORF2#3
CT SZ MD BD
hORF2#1 (SATA normalized)
CT SZ MD BD
R
el
at

iv
e
br
ai
n
L1
c
on
te
nt
0.4
2.0
1.8
1.6
1.4
1.2
1.0
0.8
0.6
p=0.0678
p=0.0295

hORF2#1 (HERVH normalized)
p=0.0298
p=0.0035
p=0.0061
0.4
2.0
1.8
1.6
1.4
1.2
1.0
0.8
0.6R
el
at
iv
e
br
ai
n

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c
on
te
nt
AlexaFluor488_NeuN
FS
C
-A
250
50
100
150
200
1051041031020
NeuN+
NeuN-
(×
1
,0
00

)
NeuN
positive
NeuN
negative
Alexa488_NeuN Merge with PI
A
B
C
D
0.4
2.0
1.8
1.6
1.4
1.2
1.0
0.8
0.6
CT SZ
0.4
2.0

1.8
1.6
1.4
1.2
1.0
0.8
0.6
CT SZ
hORF2#2 (SATA normalized)hORF2#2 (HERVH normalized)
R
el
at
iv
e
ne
ur
on
al
L
1
co
nt
en
t
R
el

at
iv
e
ne
ur
on
al
L
1
co
nt
en
t
p<0.0001p=0.0077
E
II CT 34 8:26 44.5±7.5 - 0 18:16 6.6±0.3 29.5±13.0
set diagnosis gender (F:M) age onset (yrs) suicide side (R:L)
pH PMI (hrs)n
I BD 5:8 41.5±11.2 21.9±8.9 7 7:6 6.1±0.2 31.2±15.213
I MD 5:7 45.2±10.0 33.5±11.8 6 3:9 6.1±0.2 27.8±10.512
I SZ 5:8 44.4±12.9 24.2±8.1 3 6:7 6.2±0.3 34.7±14.613
I CT 5:8 48.2±10.4 - 0 5:8 6.2±0.2 23.6±10.713
II SZ 9:26 42.6±8.5 21.3±6.1 7 18:17 6.5±0.2 31.4±15.335

Figure 1. Increase of Brain L1 Copy Number
in Schizophrenia
(A) Structure of L1 andmap of the primers. Primers
and probes are fromprevious studies (Coufal et al.,
2009; Muotri et al., 2010) or designed for this study
(Table S4). (B) Summary of the demographic vari-
ables of brain samples. (C) L1 copy number in set I.
(D) Neuronal nuclei isolation. Left: example of
NeuN-based nuclei sorting of brain cells from a
patient with schizophrenia. Right: microscopic
confirmation of isolated nuclei. The purity of each
fraction was >95% and 99.9% for NeuN+ and
NeuN�, respectively. (E) Neuronal L1 copy num-
ber in set II. In quantitative real-time PCR, L1 copy
number was measured with HERVH or SATA as
internal controls. The ratio of prefrontal cortex to
liver (for set I) or neurons to nonneurons (for set II)
was calculated and then normalized relative to the

average value of control samples. Values were
represented as open or closed diamonds as well
as box plots. The DCt values of L1 and control
probes were not significantly different between
diagnostic groups in set I or set II. p values were
determined by the Mann-Whitney U test. PMI,
postmortem interval; CT, controls; SZ, schizo-
phrenia; MD, major depression; BD, bipolar dis-
order; PI, propidium iodide. See also Tables S1
and S4 and Figures S1 and S2.
Neuron
Increased L1 Copy Number in Schizophrenia
is active in MeCP2 mouse models and patients with Rett syn-
drome, indicating a role for this mechanism in thisMendelian
dis-
order (Muotri et al., 2010). Together, these findings suggest the
Neuron 81, 306–313
hypothesis that L1 retrotransposition
may also be involved in the pathophysi-
ology of mental disorders.

In this study, we quantified L1 copy
number in genomic DNA derived from
postmortem brains of patients with major
mental disorders.We report significant in-
creases of L1 content in the prefrontal
cortex of patients with schizophrenia. To
confirm this finding, we quantified L1
copy number in neurons and nonneurons
from a second, independent patient
cohort using NeuN-based cell sorting
(Iwamoto et al., 2011; Rehen et al.,
2005; Spalding et al., 2005) and found
that L1 copy number in neurons was
increased in patients with schizophrenia.
We next quantified L1 copy number in
the animal models that are known to
disturb early neural development. These
included maternal polyriboinosinic-poly-

ribocytidilic acid (poly-I:C) injection in
mice (Meyer and Feldon, 2012; Giovanoli
et al., 2013) and chronic epidermal
growth factor (EGF) injection to infant ma-
caques (Nawa et al., 2000). We found that
genomic DNA of brains from both animal
models showed increased L1 copy number, addressing the
importance of environmental factors during perinatal and post-
natal stages. We also found that the increased L1 copy number
, January 22, 2014 ª2014 Elsevier Inc. 307
0
0.5
1.0
1.5
2.0
m5UTR mORF1 mORF2
p=0.0019
p=0.0148

CT (n=8)
Poly-I:C (n=8)
p=0.0207
R
el
at
iv
e
br
ai
n
L1
c
on
te
nt cORF2#3 cORF2#4
0.8
1.2
1.1
1.0
0.9
1.3

1.4
CT
(n
=3
)
Ha
l (n
=3
)
0.8
1.2
1.1
1.0
0.9
1.3
1.4
CT
(n
=3
)
Ha

l (n
=3
)
R
el
at
iv
e
br
ai
n
L1
c
on
te
nt
R
el
at
iv
e
br
ai
n

L1
c
on
te
nt
EG
F1
EG
F2
EG
F1
EG
F2
A B
(5s-rRNA normalized)
(5s-rRNA normalized)
Figure 2. Increase of Brain L1 Copy Number
in Animal Models
(A) Brain L1 content in thematernal poly-I:Cmodel.
p values were determined by the Mann-Whitney U
test. Values were represented as open or closed

diamonds aswell as box plots. (B) Brain L1 content
in chronic EGF or haloperidol-treated macaque
models. Error bars indicate SDs. The comparative
Ct method, with 5S-rRNA as an internal control,
was used. The ratio of prefrontal cortex to liver (for
poly-I:C model) or prefrontal gray matter to NeuN-
sorted nonneurons in white matter (for macaque
models) was calculated and then normalized
relative to the average value of control samples.
See also Table S4.
Neuron
in the neurons derived from induced pluripotent stem (iPS) cells
of schizophrenia patients with 22q11 deletion. The 22q11 dele-
tion is a well-defined genetic factor and is one of the highest
risk factors for schizophrenia, affecting about 1%–2% of
schizo-
phrenia patients (Karayiorgou et al., 2010). Finally, we
performed
whole-genome sequencing (WGS) analysis of brain and liver in

controls and patients. Comparison of brain-specific L1 insertion
sites revealed that brain-specific L1 insertion in patients is
enriched in or near genes related to synaptic function and
neuro-
psychiatric diseases. These results suggest that increased retro-
transposition of L1 in neurons, which was triggered by genetic
component and/or environmental factors at the early neural
development, could contribute to the susceptibility and patho-
physiology of schizophrenia.
RESULTS
Increased Brain L1 Content in Schizophrenia
We used postmortem prefrontal cortex samples of patients with
schizophrenia, bipolar disorder, and major depression as well as
control subjects for analysis in set I. The demographic variables
are summarized in Figure 1B. We quantified L1 copy number of
postmortem prefrontal cortex and liver in each subject by quan-
titative RT-PCR with two different internal controls, which
were
designed for human endogenous retrovirus (HERVH) and

alpha-satellite (SATA). We found a significant increase in the
brain L1ORF2 content in patients with schizophrenia (Figure
1C).
A tendency toward copy number increase was also observed in
mood disorders and in other L1 probes in schizophrenia (Fig-
ure S1 available online).
Somatic L1 retrotransposition was primarily found in neuronal
cells (Kuwabara et al., 2009). To confirm the increased brain L1
copy number in schizophrenia and address whether this copy
number increase is due to alteration of the neuronal genome,
we examined an independent prefrontal cortex sample set (set
II). We separated neuronal and nonneuronal nuclei from frozen
brains using NeuN-based cell sorting (Figure 1D) (Iwamoto
et al., 2011). NeuN is expressed in vertebrate neurons, and its
antibody can be used for labeling neuronal nuclei (Mullen et al.,
1992). We quantified L1ORF2 copy number of genomic DNA
derived from neurons (NeuN-positive nuclei) and nonneurons
(NeuN-negative nuclei) and then calculated the neuron-to-non-
neuron ratio. We found a significant increase of neuronal

L1ORF2 content in schizophrenia in two different internal con-
trols (Figure 1E). The copy number of the other L1 probes
tested
also showed significant increase in schizophrenia compared to
controls in SATA-normalized data, and similar tendency toward
copy number increase was observed in HERVH-normalized
data (Figure S1 and data not shown).
Assessment of Confounding Factors
We assessed the effect of confounding factors on L1 content
(Table S1). Among the demographic variables tested, sample
pH showed a weak correlation with L1ORF2 content in set II
but not in set I. Several variables also showed weak
correlations,
but none showed consistency across the different internal con-
trol probes or across the two different sample sets.
To consider the possible effect of antipsychotics, we exam-
ined L1 copy number in a human neuroblastoma cell line
cultured
with haloperidol or risperidone for 8 days. Both antipsychotics
did not modify the L1 copy number at their low or high concen-

trations (Figure S2). Together with the fact that the lifetime
intake
of antipsychotics, which was estimated as fluphenazine milli-
gram equivalents, did not correlate with L1 copy number in
both brain sets (Table S1), medication status did not affect our
results.
L1 Quantification in Animal Models
To assess the potential roles of environmental factors on
increased L1 copy number, we employed two different animal
models that mimic environmental risk factors that affect early
neural development. They included maternal poly-I:C injection
in mice and chronic EGF injection to neonatal macaques. The
poly-I:C, which mimics viral double-stranded RNA, injection to
pregnant mice induces elevated maternal immune activation,
and the offspring is known to show schizophrenia-like
behavioral
alterations such as impairments of prepulse inhibition and social
behavior at the later stage (Meyer and Feldon, 2012). Pregnant
mice received a single intraperitoneal injection of poly-I:C. L1

copy number in the prefrontal cortex of offspring was tested at
postnatal day 21. We found that significant elevation of L1 copy
number at all the tested probes compared to controls (Figure
2A).
We then examined the L1 copy number in macaques treated
with EGF during neonatal period. Perinatal and postnatal
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Figure 3. L1 Content in Neurons Derived from iPS Cells of
Schizophrenia Patients with 22q11 Deletions
The comparative Ct method, with SATA as an internal control,
was used. The ratio of NeuN-sorted neurons to nonneurons was
calculated and then normalized
relative to average value of control samples. Error bars indicate
SDs. See also Table S4 and Figure S3.
Neuron
perturbation of EGF is known to evoke schizophrenia-like phe-
notypes, including deficits in prepulse inhibition, latent
inhibition,
social interaction, and working memory, in adulthood (Nawa

et al., 2009, 2000).The neonatal macaques (n = 2) subcutane-
ously received EGF for seven times over 11 days. After 4 and 7
years from treatment, L1 copy number in the prefrontal cortex
was tested. In addition, chronic haloperidol-treated macaques
(n = 3) were also tested. Due to unavailability of other tissues,
we isolated nonneuronal nuclei from frozen white matter and
calculated the grey matter-to-nonneuron ratio in each subject.
Although statistical approach could not be applied, we observed
increase of L1 copy number in EGF-treatedmacaques, but not in
the haloperidol-treated macaques, compared to controls (Fig-
ure 2B). Taken together, these results suggest that early
environ-
mental factors play important roles in the L1 content in the
brain.
We further confirmed that chronic haloperidol treatment did not
influence L1 copy number in this model.
L1 Quantification in the iPS Cells of Schizophrenia
Patients with 22q11 Deletion
We next assessed the importance of genetic risk factor on the
L1

copy number in brain. We quantified L1 copy number in the
neu-
rons derived from iPS cells of schizophrenia patients with
22q11
deletion (n = 2) as well as controls (n = 2) (Figure S3). The iPS
cells
were established from the fibroblasts according to the
previously
developed method (Imaizumi et al., 2012; Takahashi et al.,
2007;
M.T., unpublished data). To estimate the L1 copy number, we
used two independently established iPS cell lines per patient.
After induction of neuronal cells (Imaizumi et al., 2012), we
iso-
lated neuronal nuclei by NeuN-based sorting (Figure S3). We
then examined L1 copy number and calculated the neuron-to-
nonneuron ratio. Compared to controls, we observed consistent
increase of L1 copy number in iPS cell-derived neurons of
patients with schizophrenia with 22q11 deletion (Figure 3).
These
results suggest that the well-defined strong genetic risk factor
also plays an important role in the L1 content in the brain.

Identification and Comparison of Brain-Specific L1
Transposition
We next performed WGS of brain and liver DNA from same
sub-
jects by self-assembling DNA nanoarray technology (Drmanac
et al., 2010). For this experiment, schizophrenia patients (n = 3)
and control subjects (n = 3) were selected to match age, PMI,
gender, brain pH, and race from set I. Selected patients ex-
hibited increased L1 content by quantitative RT-PCR assay,
compared to average L1 content of the controls and selected
control subjects. The WGS metrics and identified variations
were summarized in Table S2. Distribution of the detected mo-
bile elements was almost equal between the tissues and across
subjects, and over the half of the identified elements was related
Neuron 81, 306–313, January 22, 2014 ª2014 Elsevier Inc. 309
Control
Term Count p value FE
height 4 0.0132 7.7
scoliosis 3 0.0316 10.3
Schizophrenia
Term Count p value FE
schizophrenia;

bipolar disorder
schizoaffective disorder; 5 0.0125 5.2
schizophrenia 29 0.0135 1.6
hypertension 20 0.0194 1.7
bipolar disorder 13 0.0373 1.9
A
Control
Term Count p value
GO:0005856~cytoskeleton 74 5.92E-04
GO:0005509~calcium ion binding 56 0.0031
GO:0005930~axoneme 9 0.0095
GO:0035085~cilium axoneme 7 0.0289
GO:0003779~actin binding 26 0.0322
GO:0044425~membrane part 240 0.0387
GO:0016010~dystrophin-associated glycoprotein complex 6
0.0405
Schizophrenia
Term Count p value
GO:0045202~synapse 57 3.09E-09
GO:0030054~cell junction 64 8.57E-06
GO:0044459~plasma membrane part 187 1.49E-05
GO:0004674~protein serine/threonine kinase activity 58 2.27E-
05
GO:0044456~synapse part 38 4.44E-05
GO:0004672~protein kinase activity 72 7.45E-05
GO:0030554~adenyl nucleotide binding 147 1.09E-04
GO:0005856~cytoskeleton 126 1.19E-04
GO:0005488~binding 820 1.25E-04
GO:0006468~protein amino acid phosphorylation 76 1.75E-04
GO:0006796~phosphate metabolic process 100 1.98E-04
GO:0006793~phosphorus metabolic process 100 1.98E-04
GO:0016773~phosphotransferase activity, alcohol group as

acceptor 80 2.31E-04
GO:0001882~nucleoside binding 148 2.52E-04
GO:0005524~ATP binding 138 2.63E-04
GO:0001883~purine nucleoside binding 147 2.77E-04
GO:0032559~adenyl ribonucleotide binding 139 3.43E-04
GO:0017076~purine nucleotide binding 169 4.19E-04
GO:0000166~nucleotide binding 190 0.0011
GO:0014069~postsynaptic density 17 0.0011
GO:0032553~ribonucleotide binding 161 0.0012
GO:0032555~purine ribonucleotide binding 161 0.0012
GO:0043167~ion binding 323 0.0015
GO:0016043~cellular component organization 204 0.0018
GO:0043169~cation binding 318 0.0021
GO:0016310~phosphorylation 83 0.0021
GO:0046872~metal ion binding 315 0.0025
GO:0016301~kinase activity 85 0.0032
GO:0005737~cytoplasm 489 0.0037
GO:0008092~cytoskeletal protein binding 58 0.0045
GO:0007155~cell adhesion 74 0.0049
GO:0022610~biological adhesion 74 0.0052
GO:0019898~extrinsic to membrane 54 0.0053
GO:0043687~post-translational protein modification 108 0.0169
GO:0030030~cell projection organization 45 0.0219
GO:0005509~calcium ion binding 88 0.0234
GO:0015629~actin cytoskeleton 33 0.0439
B
insertion site CT1 CT2 CT3 average ratio
intergenic (low) 0.64 0.76 0.69 0.69
intragenic (low) 0.36 0.24 0.31 0.31
intron (low) 0.90 1.00 0.99 0.96
exon (low) 0.10 0.00 0.01 0.04
intergenic (high) 0.65 0.81 0.69 0.72
intragenic (high) 0.35 0.19 0.31 0.28
intron (high) 0.88 1.00 0.99 0.96

exon (high) 0.12 0.00 0.01 0.04
C
90 0.0473
GO:0045211~postsynaptic membrane 21 0.0487
insertion site SZ1 SZ2 SZ3 average ratio
intergenic (low) 0.72 0.63 0.58 0.64
intragenic (low) 0.28 0.37 0.42 0.36
intron (low) 0.99 0.98 0.98 0.98
exon (low) 0.01 0.02 0.02 0.02
intergenic (high) 0.74 0.63 0.57 0.65
intragenic (high) 0.26 0.37 0.43 0.35
intron (high) 0.99 0.98 0.98 0.98
exon (high) 0.01 0.02 0.02 0.02
Control
Schizophrenia
GO:0016772~transferase activity, transferring phosphorus-
containing groups
Figure 4. Insertion Site, Gene Ontology, and Disease
Association Analyses
(A) L1-insertion site analysis. Proportion of intergenic and
intragenic L1 insertion and that of intronic and exonic L1
insertion are given. The low and high mean
estimated proportions based on both less and stringent criteria
are given. Note that ratios are not significantly different
between patients and controls. (B) Gene
ontology analysis. p values indicate Bonferroni-corrected

modified Fisher’s exact test p value. The terms showing p <
0.05 are shown for both groups. (C)
Disease-association analysis. p values indicate noncorrected
modified Fisher’s exact test p value. FE, fold enrichment. In
both analyses, gene lists generated by
the stringent criteria were used. See also Tables S2 and S3 and
Figure S4.
Neuron
to the L1-Hs (Figure S4). Among the detected mobile element
insertion sites in each sequenced sample, we first identified
brain-specific L1 insertions in each subject (Tables S2 and S3).
Although the total number of brain-specific L1 insertion tended
to be higher in schizophrenia patients, this was not statistically
significant, most likely due to the limited sample size and high
interindividual variation. We then compared genomic locations
of the insertion sites of brain-specific L1 between patients and
controls (Figure 4A). The inter-to-intragenic L1 insertion ratio
as
well as exonic-to-intronic L1 insertion ratio did not differ
between
patients and controls. We then compared the affected genes by

brain-specific L1 insertion by gene ontology approach. This
analysis revealed that the number of enriched terms is higher
in schizophrenia than controls, in spite that the number of
brain-specific L1 insertions did not significantly differ. We
found
that neuronal function-related terms such as synapse and
protein phosphorylation are clearly overrepresented in schizo-
phrenia compared to controls (Figure 4B). In addition, disease-
association analysis revealed that affected genes in patients
are specifically enriched in terms related to schizophrenia and
bi-
polar disorder, while those in controls are enriched in nonneur-
opsychiatric terms such as height and scoliosis (Figure 4C).
These results were consistently confirmed when we used less
stringent definition of brain-specific L1 insertion (Figure S4).
In
Neuron
addition, enrichment of the L1-inserted genes to the terms

related to neuropsychiatric disorders in schizophrenia was also
detected by the ingenuity pathway analysis (IPA) (Figure S4).
DISCUSSION
We report that the neuronal genome of schizophrenia contains
higher copy number of a retrotransposon, L1. To validate this
finding, we utilized iPS cells from patients with schizophrenia
carrying the 22q11 deletion and observed an increase in L1
copy number in iPS cell-derived neurons. Moreover, using
WGS, we found that L1 preferentially inserted into genes
related
to synaptic functions and schizophrenia. Animal model studies
showed that environmental factors related to infection or
inflam-
mation that disturbs early neurodevelopmental processes
increase L1 copy number in the brain. Collectively, these results
suggest that hyperactive L1 retrotransposition into critical
genes during neural development, triggered by genetic and/or
environmental factors, contribute to the pathophysiology of
schizophrenia. Our results significantly expand the range of

neuropsychiatric illnesses linked to aberrant L1 retrotransposi-
tion, from Mendelian disease patients with MECP2 mutations
in Rett syndrome (Muotri et al., 2010) and ATM mutations in
ataxia telangiectasia (Coufal et al., 2011) to schizophrenia, a
complex mental disorder.
The observed increase of L1 content in schizophrenia was not
due to, or modulated by, biological or experimental artifacts,
because changes were measured in two independent patient
cohorts and each result was confirmed with two different
internal
controls. Although the L1 region showing significant increases
differed between the two brain sets, this is attributable to cohort
differences amplified by the strict threshold we employed.
Actu-
ally, a significant increase of L1 content was widely observed in
all probes in the SATA-normalized data in set II, where
neuronal
L1 copy number was directly examined (Figure S1). In addition,
from the data analysis utilizing lifetime intake of antipsychotics

of patients, and from the cell culture and macaque experiments,
we conclude that antipsychotics do not affect L1 copy number
in
the brain. A significant increase was also observed in patients
with mood disorders in one internal control in set I (Figure 1C).
Future work will clarify whether there are L1 content increases
in other mental disorders using larger and/or stratified patient
cohorts.
L1 retrotransposition has been detected during adult neuro-
genesis in the rat hippocampus, indicating that neural progenitor
cells retain retrotransposition activity even in adult stages
(Muotri
et al., 2009). However, we analyzed potential confounding fac-
tors, including age, age of onset, and duration of illness, and
did not observe any significant correlation with L1 copy number
in the brain. The transcript level of L1 in adult brain sample
was
also increased in patients compared to controls (data not
shown). However, elevated expression is unlikely to contribute
to increase of L1 copy number in patients, as significant

increase
of L1 transcripts was detected only in the 50 region of L1 such
as
50 UTR and ORF1. These results suggest that L1 copy number
does not globally increase with aging and that the variation of
L1 copy number in patients is probably confined to early neuro-
developmental stages, at least in the prefrontal cortex. This
prediction would be consistent with the neurodevelopmental hy-
pothesis of schizophrenia, where abnormalities during critical
early periods of brain development may trigger the later appear-
ance of clinical symptoms (Bloom, 1993; Murray et al., 1992;
Weinberger, 1987).
In Rett syndrome, increased L1 copy number in human brain
was linked to mutations in MECP2 (Muotri et al., 2010) and
MeCP2 knockout mice also showed increased L1 content
(Muotri et al., 2010). It has also been suggested that SOX2 and
MECP2 regulate L1 transcription in neurons (Muotri et al.,
2005; Yu et al., 2001). However, we did not observe a
significant
correlation between MECP2 or SOX2 expression and brain L1
content, by using the previously performed gene expression

analyses on the same sample sets (Iwamoto et al., 2004, 2005)
(data not shown). In addition, patients with high levels of L1
copy number (two schizophrenia and one major depression in
set I, and two schizophrenia patients in set II) did not show
altered MECP2 or SOX2 expression levels (data not shown).
These findings suggest that the molecular mechanism of
increased L1 in schizophrenia is different from Rett syndrome.
In this study, we found that both early environmental and well-
defined strong genetic factors for schizophrenia are involved in
the increase of L1 copy number in the brain. A recent study
using
the poly-I:C model indicated that the offspring of this model
had exacerbated schizophrenia-like phenotypes, if they were
exposed to environmental stress during puberty, suggesting
that early environmental factors can lower the threshold for
onset
of schizophrenia (Giovanoli et al., 2013). Therefore, increased
L1
insertions induced by environmental factors may increase the

susceptibility to schizophrenia by disrupting synaptic and
schizophrenia-related genes in neurons, rather than being a
direct cause of the disease. On the other hand, the pathological
consequences of increased L1 content in neurons derived from
iPS cells of schizophrenia patients with 22q11 deletions remain
unclear. We chose patients with 22q11 deletions to examine
L1 dynamics where there is a well-defined strong genetic risk
for schizophrenia. InMeCP2-knockoutmice, Rett-like behavioral
abnormalities could be rescued by the re-expression of wild-
type MeCP2 at both young and adult stages (Cobb et al.,
2010; Ehninger et al., 2008), suggesting that L1 content itself
may not be directly causal to disease phenotypes but instead
modulate phenotypic variability among patients (Muotri et al.,
2010). Similarly, we speculate that the L1 increase in schizo-
phrenia patients with 22q11 deletions is likely to modulate phe-
notypes of schizophrenia rather than a direct cause, because
many genes related to schizophrenia, such as TBX-1, SEPT5,
COMT, and PRODH, are located within the deletion (Hiroi

et al., 2013; Karayiorgou et al., 2010). Nevertheless, our
findings
will facilitate further studies of the mechanism of increased L1
retrotransposition associated with schizophrenia.
Our WGS analysis could not detect increased brain-specific
L1 insertions in schizophrenia; however, we found that L1
inser-
tions were more frequent in genes for synaptic function and
schizophrenia relative to controls. Evrony et al. cloned one L1
insertion event from 300 single neurons and showed that 2 of
83 cortical neurons from an individual had this insertion, but
detection of such a low level mosaic insertion in bulk brain
tissue
of the same individual was difficult and needed optimization
Neuron
(Evrony et al., 2012). Thus, rare L1 insertion events could be
missed in our WGS analysis. Apart from L1, nonautonomous
ret-

rotransposons such as Alu and SVA also show an increased
copy number in the brain, possibly via the aid of L1ORF
products
(Baillie et al., 2011) and their copy number might also be
increased in patients. Further studies on the neuronal genome
of patients with mental disorders, and supporting mechanistic
evidence from animal and cellular models, may establish a
broader role for instability of neural genome in the pathophysi-
ology of schizophrenia. We expect that our findings will
promote
the further study of genomic instability in disease etiology due
to
L1 retrotransposition in brain development.
EXPERIMENTAL PROCEDURES
Postmortem Samples
Postmortem brain and liver samples were obtained from the
Stanley Medical
Research Institute. The demographics are summarized in Figure
1B and are
described at the web site (http://www.stanleyresearch.org/).
Ethics commit-

tees of RIKEN and the University of Tokyo Faculty of Medicine
approved the
study.
Animal Models
Animal experiments were performed in accordancewith the
NIHGuidelines for
the Care and Use of Laboratory Animals and guidelines of
relevant facilities.
For poly I:C model, pregnant mice (C57BL/6) received either a
single intraper-
itoneal injection of poly-I:C (2 mg/ml, Sigma-Aldrich)
dissolved in PBS
(20 mg/kg) or an equivalent volume of PBS at embryonic day
9.5. At postnatal
day 21, tissues were dissected from pups. For macaque models,
cynomolo-
gusmonkeys (Macaca fascicularis) (4 years old; all males) were
given oral halo-
peridol (0.25–0.5 mg/kg; Wako Pure Chemical Industries) or
vehicle for
2 months (Shibuya et al., 2010). After transiently separating
two male monkey
neonates (2 weeks old) from dams, neonates received
subcutaneous admin-

istration of human recombinant EGF (0.3 mg/kg, Funakoshi) for
seven times
over 11 days and then quickly returned to their dams.
Preliminary behavioral
assessment of the EGF-treated monkeys was performed at ages
of 4 and 6
years and reported (Nawa et al., 2009). These monkeys were
sacrificed at
the age of 4 and 7 years with the overdose of pentobarbital (26
mg/kg;
65 mg/ml). Experiments were subjected to review by the Ethical
Committee
of Shinn Nippon Biomedical Lab.
iPS Cells
All procedures for skin biopsy and iPS cell production were
approved by the
Keio University School of Medicine ethics committee and
RIKEN ethics com-
mittee. The 201B7 iPS cells were kindly provided by Dr.
Yamanaka (Takahashi
et al., 2007). For the control WD39, a skin-punch biopsy from a
healthy
16-year-old Japanese female obtained after written informed

consent was
used to generate iPS cells (Imaizumi et al., 2012). 22q11.2
deletion syndrome
iPS cells (SA001 and KO001) were generated from a 37-year-
old Japanese
female patient (Toyosima et al., 2011) and a 30-year-old
Japanese female
patient, respectively, using the same method used to generate
the WD39
(M.T., unpublished data). 22q11 deletion was characterized by
the CGH array
analysis (Figure S3). Production and maintenance of iPS cells
were performed
according to the previous studies (Imaizumi et al., 2012;
Takahashi et al.,
2007). All the iPS cells and differentiated neuronal cell lines
were characterized
with immunofluorescence staining and their morphologies
(Figure S3).
L1 Copy Number Estimation
We performed either Taqman-based quantitative real-time PCR
according to
Coufal et al. (2009) with minor modifications (100 or 500 pg
DNA as starting

material and single amplicon analysis) or SYBR-Green-based
quantitative
real-time PCR according to Muotri et al. (2010). SYBR-Green
assay was
performed using 500 pg DNA and Power SYBR Green PCR
Master Mix (Life
Technologies). Primers, probe location, and reaction chemistry
are listed in
Figure 1A and Table S4. Quantification was performed in
triplicate. A nonpara-
metric Mann-Whitney U test was employed for two group
comparison and
p < 0.05 was considered significant.
Whole-Genome Sequencing
WGS of brain and liver samples from controls and
schizophrenia patients was
performed by Complete Genomics, with the paired-end library
preparation
and sequencing-by-ligation using self-assembling DNA nanoball
(DNB)
(Drmanac et al., 2010). Data process, mapping, and detection of
variations
were performed using the software developed by the Complete

Genomics
(version 2.2.0.26 and format version 2.2). Among the
detectedmobile insertion
elements, we compared the genomic location of L1 insertion
between brain
and liver within an individual and identified brain-specific L1
insertions.
Further experimental details are available in the Supplemental
Experimental
Procedures.
SUPPLEMENTAL INFORMATION
Supplemental Information includes Supplemental Experimental
Procedures,
four figures, and four tables and can be found with this article
online at
http://dx.doi.org/10.1016/j.neuron.2013.10.053.
ACKNOWLEDGMENTS
This work was supported in part by the Grant-in-Aid for
Scientific Research on
Innovative Areas (Unraveling the microendophenotypes of
psychiatric disor-
ders at the molecular, cellular, and circuit levels) from the
Ministry of Educa-

tion, Culture, Sports, Science and Technology (MEXT) to T.Y.,
H.N., T.K.,
and K.I., and a Grant-in-Aid from Ministry of Health, Labour
and Welfare to
T.K. This work was also supported by JST, CREST to T.K. and
by JST,
PRESTO to K.I. This work was also supported in part by
Leading Project for
Realization of Regenerative Medicine from MEXT and
‘‘Funding Program for
World-Leading Innovative R&D on Science and Technology’’ to
H.O., and by
the ‘‘Development of biomarker candidates for social behavior’’
carried out un-
der the Strategic Research Program for Brain Sciences
fromMEXT to T.Y. and
K.K. This work was also supported in part by the Collaborative
Research Proj-
ect of the Brain Research Institute, Niigata University.
Postmortem samples
were donated by the Stanley Medical Research Institute,
courtesy of
Drs. Michael B. Knable, E. Fuller Torrey, Maree J. Webster,
and Robert H.

Yolken. We thank Tomoko Toyota and Atsuko Komori-Kokubo
at RIKEN BSI
for their technical assistance. We also thank Kenji Ohtawa at
Research Re-
sources Center at the RIKEN BSI for the cell-sorting analysis.
M.B., F.S.,
and K.I. belong to the Department of Molecular Psychiatry,
which is endowed
by Dainippon Sumitomo Pharma and Yoshitomiyakuhin. H.O. is
a scientific
consultant for San Bio, Eisai, and Daiichi Sankyo. T.K.
received a grant from
Takeda Pharmaceutical. These companies had no role in study
design, data
collection and analysis, decision to publish, or preparation of
the manuscript.
Accepted: October 18, 2013
Published: January 2, 2014
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Increased L1 Retrotransposition in the Neuronal Genome in
SchizophreniaIntroductionResultsIncreased Brain L1 Content in
SchizophreniaAssessment of Confounding FactorsL1
Quantification in Animal ModelsL1 Quantification in the iPS
Cells of Schizophrenia Patients with 22q11
DeletionIdentification and Comparison of Brain-Specific L1
TranspositionDiscussionExperimental ProceduresPostmortem
SamplesAnimal ModelsiPS CellsL1 Copy Number
EstimationWhole-Genome SequencingSupplemental
InformationAcknowledgmentsReferences

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