Monday, November 18, 2013

Concussion Healing Aided by Menstruation



Photo via askchiq.com  
               When you think of sports related concussions you probably think of men’s football, rugby, ice hockey, and other aggressive sports but, many wouldn’t jump to females suffering many concussions due to sports injuries as well. Well according to Dr. Bazarian at the University of Rochester Sport Concussion Clinic about 70% of the head injuries patients he sees are females. Women typically show longer recovery times than do men when faced with a head injury but, menstrual cycles could play a role in just how long.
                After receiving a head injury sex hormones are often immediately changed from the normal levels. This is due to the pituitary gland in the brain which can be shut down by a hit to the head. The pituitary gland is part of the endocrine system which releases several hormones such as luteinizing hormone (LH) and follicle stimulating hormone (FSH) which are both important to the regular menstrual cycle of females (Discovery Communications, LLC. , 2013). The pituitary is often called
Photo via WebMD
the "master gland” because it can influence a majority of all the other glands in the body; it has the ability to manufacture and store hormones that can be released, causing the stimulation of other glands (Discovery Communications, LLC, 2013). Researchers have determined that if the pituitary gland no longer produces and releases FSH and LH then the levels of estrogen and progesterone in the body will drastically drop (University of Rochester Medical Center, 2013). FSH and LH are an important part of the menstrual cycle and greatly affect the release of progesterone, as can be seen in the chart shown below.

                The hormone progesterone is a hormone in the human body that is produced in the ovaries, in the placenta after implantation of a fetus, and by the adrenal glands (National Women's Health Resource Center, 2013). Progesterone is important in preparing the body for conception and is also responsible for maintaining a regular monthly menstrual cycle (National Women's Health Resource Center, 2013). As is shown in the diagram, when the levels of progesterone are high the uterine lining is thickening, preparing for implantation and if an egg is not fertilized and implanted the levels of progesterone quickly drop and the lining will be broken down and removed via menses (National Women's Health Resource Center, 2013). The menstrual cycle is broken into the two weeks prior to menstruation and the two weeks after, including the seven day menses. The point you are in this cycle proves to be viable information for females recovering from concussion and other head trauma related injuries.


 
      Bazarian and his team took 144 women from age 18 to 60 that had just experienced a head injury within the past four hours. These women gave blood samples as soon as they arrived and their progesterone levels were calculated determining the phase of the menstrual cycle they were currently in (University of Rochester Medical Center, 2013). Based on progesterone levels all 144 females were placed into three groups: the first was women who were in the premenstrual phase; their progesterone levels were high. The second group tested low for progesterone, which indicated they are postmenstrual (University of Rochester Medical Center, 2013). The third and final group was those who reported being on birth control pills which have synthesized progesterone hormones and females taking birth control pills receive a constant flow of hormones (University of Rochester Medical Center, 2013).
                The researchers and Bazarian came up with the “withdrawal hypothesis” before proceeding with any additional follow-up tests. This hypothesis stated that if a woman suffers a concussion during the premenstrual period her progesterone levels are high and after the injury they are drastically dropped leaving her with symptoms such as headaches, nausea, dizziness, and other uncomfortable side effects (University of Rochester Medical Center, 2013). But, if she suffers a concussion after her menstrual cycle her progesterone levels are already drastically reduced naturally so the effects previously stated won’t apply to these females because the progesterone isn’t dropping dramatically (University of Rochester Medical Center, 2013). This should cause females in the premenstrual group to suffer longer after their injury while postmenstrual females recover quicker.
After one month these same 144 females were brought back to do follow-up testing to see if the different phases of menstruation and progesterone levels have effects on the slow recovery time females endure after head injury. Standardized tests were used to assess the women for their concussion recovery and their quality of life; they were asked questions about their mobility, daily activities, and the pains they experience (University of Rochester Medical Center, 2013). The women who were in the premenstrual phase were twice as likely to score worse on these tests as the postmenstrual women and those on the pill. Women in the birth control group has the highest scores overall than the other groups. These results concur with Bazarian’s “withdrawal hypothesis” but further testing needs to be done to conclude that birth control pills are even more advantageous to female athletes. Birth control pills are already popular among female athletes because of the predictable periods and the added protection for bones; if this data is another plus for birth control pills it will be one more reason for female athletes to use them.
AskChiq. (2010, December 08). Concussion symptoms differ between girls and boys. Retrieved from http://askchiq.com/fitness_health/5588432 
Discovery Communications, LLC. (2013). What does the pituitary gland do?. Retrieved from http://health.howstuffworks.com/human-body/parts/what-does-pituitary-gland-do.htm
National Women's Health Resource Center. (2013). Progesterone overview. Retrieved from http://www.healthywomen.org/condition/progesterone
University of Rochester Medical Center (2013, November 13). Menstrual cycle influences concussion outcomes. ScienceDaily, Retrieved November 16, 2013, from http://www.sciencedaily.com/releases/2013/11/131113130033.htm
WebMD. (2010, July 23). Pituitary gland. Retrieved from http://www.webmd.com/brain/pituitary-gland
Womack, L. (2013). Birth control and the female athlete. Retrieved from http://www.sportsmd.com/SportsMD_Articles/id/309.aspx
 

Monday, November 11, 2013

Autism and my future

A Child's Journey out of Autism by Leeann Whiffen
          Autism Spectrum Disorder (ASD) is a topic that is gaining popularity and backing through research. Countless studies are trying to crack the genetic mystery of where this disorder originates or what causes it. Are there factors prenatally that affect the outcome of autistic children? My interest in this particular disorder originated from a book I read a few years ago titled, A Child's Jorney out of Autism by Leeann Whiffen about her personal experience with her son Clay who was diagnosed with an autism spectrum disorder. The story was very inspirational about her triumphs and her struggle to get him better in every way and eventually off the spectrum entirely.

Autism Spectrum Disorder consists of a wide variety of symptoms and complex brain developmental disorders. Each child with ASD has their own set of symptoms and their own unique deficits, whether it be with social interactions, difficulties getting their words and feelings across, or performing acts repeatedly without cause (NINDS, 2013). Because of the complexity of the brain and the individualized deficits experienced within each individual makes getting to the bottom of the disorder harder than anyone anticipated. In the book, Clay showed severe ASD symptoms that included  gastrointestinal problems (GI); which according to a recent study at the UC Davis MIND Institute has found to be a common side effect to the behavioral symptoms of ASD (Science Daily, 2013). Behaviors including the social deficits, repetitiveness, and irritability are linked to the GI upsets including constipation, difficulty swallowing food and food sensitivities. In Clay’s case he was put on a strict gluten-free, daily-free diet and his food intake was monitored very closely by his mother, these changes weren’t easy to adjust to for Clay but, they eventually paid off when he was removed from the spectrum. Although Clay’s story is a success, there is still no research to back-up the research done at the UC Davis MIND Institute about the treatments of these prevalent GI problems in ASD children, the research only focused on the fact that there is a correlation between them.  
How common is ASD you may be wondering? According to statistics from the U.S. Center for Disease Control and Prevention (CDC) currently 1 in 88 children born into the U.S. are diagnosed with an ASD (NINDS, 2013).  
Why does this interest me so much? I plan on becoming an occupational therapist for children in my career and many children that receive occupational therapy (OT) in the school system suffer from an autism spectrum disorder. In my experience shadowing at local elementary schools over the past year I’ve seen the effects of OT in the students that receive it. They play games and do crafts and get one-on-one attention that they might not get at home. These children love going to OT and they feel special that they get to go during class and do these things. I’ve always been the kind of person that tries everything to make someone else feel better, whether it’s saying the thing they need to hear, to being there for support, or anything else they need and experiencing the feeling that you made someone else feel better has always been something I strive for.
From the time I was a little girl all the way through high school I thought I wanted to teach elementary school kids, but when someone told me I would be good at doing occupational therapy I looked into it and I thought that person was right. I can’t wait until the day that my goals are achieved, the day I become an occupational therapist and start to make my mark on the World. I hope to experience the joy of helping children and hopefully getting the chance to specialize with those children with autism spectrum disorders. The research being done on the entire spectrum hopefully will provide some answers that will be of help to me in future years. For now I am simply fascinated by stories like Leeann, who gave up everything to help her son come off the autism spectrum.

         I highly recommend this book for anyone of any age it is truly inspiring.


NINDS. (2013, November 06). Autism fact sheet. Retrieved from http://www.ninds.nih.gov/disorders/autism/detail_autism.htm
Science Daily. (2013, November 06). Children who have autism far more likely to have tummy troubles. Retrieved from http://www.sciencedaily.com/releases/2013/11/131106202237.htm
The Puzzle Place. (2005). The puzzle place foundation. Retrieved from

              http://www.thepuzzleplace.com/sponsors.html

Whiffen, L. (2013). Leeann whiffen. Retrieved from http://leeannwhiffen.com/

 

Monday, November 4, 2013

Sea turtles facing extinction

Photo Credit Morris, 2012
                               
            Throwing a soda bottle out the window, tossing an empty bag aside on a mid-morning stroll actions that both mean nothing at the time; you're just getting rid of garbage you don't want to carry. We've all done it before but not many of us think further into the future at what those simple actions will lead to. Before learning about sea turtles I wouldn't have thought twice about it but there are serious consequences for these careless acts.

            When it comes to sea turtles pollution is a serious problem for them as a species especially plastics that make their way into our oceans. Sea turtle often ingest these plastics mistaking them for food sources such as jellyfish, which often get mistaken for plastic bags floating in the water. When plastics such as this get digested there are chemicals released inside the turtle that produce gases giving turtles a symptom termed "bubble-butts" because they float (Carr, 2011) This side effect is often lethal to the sea turtles because they're rear ends float and they suffocate not being able to reach the surface (Carr, 2011).

            Plastics and chemicals aren't the only new problems sea turtles have to face, there are several man-made factors influencing their lives and making it a struggle. A major problem is the rise of coastal developments along beaches where sea turtles nest. There are a multitude of side effects from these establishments; one being the artificial light that is overcast onto the beach induces “false crawls” for the female turtle and they often don't nest on these beaches and choose less optimum places to nest (Magnuson, 1990). These lights also disorient hatchlings as they try to make their way to the sea, they don't see well on land and look for the horizon lights of the sea and when the artificial lights are brighter than the sea lights they sometimes crawl inward and get into traffic (Magnuson, 1990) The influx of people to the coastal areas also increases the amount of activity on the beach both during the day and during the night. Female sea turtles are very particular about where they nest and the conditions they need to have in order to do so. These include a completely quiet, dark place where they don't feel threatened or in danger (Carr, 2011). Humans on the beach at night don't allow the female sea turtle to feel safe so they retreat back to another beach to nest. Another consequence of humans living along the coast is the occurrence of beach furniture left behind by them (Carr, 2011). These cause unnatural obstacles for both female turtles trying to nest as well as the hatchlings trying to journey to sea and can even cause strangulation or predation to occur.

            Along with humans come they're domesticated pets like dogs and cats; these animals often roam free along the coast and become predators to sea turtles. They often wander along the coast during the night and come across a nesting female either in the process or making her way to a nesting spot. The presence of the dog, for example, will either make her retreat back or get attacked (Carr, 2011). With the success of finding the turtle the animal will return to that beach in hopes of finding another turtle and thus the beach is no longer a viable nesting ground for that specific turtle. The scent of trash left behind by humans during their beach lounging during the day invites creatures like raccoons, which aren’t natural predators of sea turtles, to the beach in search of food. Like with domesticated animals the raccoons will often find the turtles and continue coming back (Carr, 2011).

            Humans are a big problem when it comes to the sea turtles demise, but the good news is that human activities can be monitored and resolved. There’s no easy or quick fix for this problem but there is a slight chance of hope because the problem isn’t of natural causes. If the extinction of sea turtles was of natural causes due to say global climate change there would be nothing us humans could do for them. Thankfully there are already conservationists working to get laws enforced and countless turtle hospitals taking in sick turtles.

Visit the following links to learn more about sea turtle conservation and sea turtle hospitals.        

                                          Conservation Efforts         Turtle Hospital

 

 

Carr, A. (2011). Information about sea turtles, their habitats, and threats to survival. Retrieved from http://www.conserveturtles.org/
 
Magnuson, J. (1990). Decline of sea turtles. Washington D.C.: National Academy Press.

Morris, H. (2012, March 13). Sea turtle hatchling. Retrieved from http://calibermag.org/articles/should-animal-captivity-be-allowed/sea-turtle-hatchling/\


 

Estrogen gives females an advantage with stress


Photo Credit Greenfieldboyce, 2011
Females are superior to males; a fact that women have been scouring to prove for decades, even if for their own satisfaction or to just rub in it the faces of men. But, there may be certain key benefits for those of us with the ability to produce estrogen. A recent study from the University of Buffalo has been working to determine the protective effects of estrogen in the brains of female rats in response to stress. Overall females tend to show a better response or less of an adverse effect to stress and the underlying factor may be estrogen. You may say, “males have the ability to produce estrogen as well”, and although that statement is true the amounts of which are produced in both are drastically different. But, if males could produce or be given a safe way to produce or maintain estrogen in their brains they may have the same reaction to stress.

The original article "Estrogen protects against the detrimental effects of repeated stress on glutamatergic transmission and cognition" outlines the entire procedure done on the rats for this experiment. The researchers used several different methods of recording data from the rats being examined; the first was simple behavior testing. The rats were subject to temporal order recognition memory (TORM) tasks, which consisted of two sample phases and one test phase (Wei & Yuen, 2013). During the sample phases two plastic objects, of the same height and material, were shown to the rats and the rats were allowed to check out (Wei & Yuen, 2013). In each phase there were two different objects, but in the final test phase one object from the first phase and another from the second phase were used to test the rats short-term memory (Wei & Yuen, 2013). If everything is in working order within the rats brain, the rats will spend more time checking out the object from phase one because it has been the longest since it has seen it (Wei & Yuen, 2013). Another test used was actual slices of the experimental rats’ brains to examine closer into the physiological effects of stress.

The stressors of the experiment were preformed two-hours daily for five to seven days with the last happening about a day before test were going to be done on the individual rat (Wei & Yuen, 2013). The rats were placed in cylinder containers that were just barely big enough to contain the rat with minimal room to maneuver around within it (Wei & Yuen, 2013). This stimulated a stressful response in the rats given the confines of the space they were limited to compared to the space they were used to. After the stressed rats came out of the last round in the small containers a day was used as s rest period before the TORM tests began.

Female rats showed much better responses in all categories studies compared to male mice so the researchers wanted to look into the effects of estrogen specifically. The repeated stressors imposed on females showed to be the same as males when their estrogen signaling was blocked and the male rats exhibited behavior similar to females when their estrogen signaling was activated (Wei & Yuen, 2013). This experimentation shows a linkage between estrogen and the effects of stress.

The biology behind the estrogen receptors in the brain is focused on the glutamate receptors in the prefrontal cortexregion of the brain which is the outermost layer of brain matter directly behind the forehead, or frontal bone (Prefrontal cortex, 2013). This area is very important in complex cognitive processes such as memory and emotion (Prefrontal Cortex, 2013). Glutamate receptors are located in the prefrontal cortex and the neurotransmitter glutamate is essential to the central nervous system because of their properties as an excitatory transmitter (Danbolt, 2001). The normal functioning of glutamate neurotransmitters is important to human memory, cognition, and learning which are all very important to human activities (Danbolt, 2001).

In the study discussed researchers found that glutamate receptors in the prefrontal cortex of female rat brain slices examined were completely intact, but not intact in males (University at Buffalo, 2013). This gives researchers clues that the glutamate receptors are targets of stress causing a loss of function in that area which houses memory and cognition (University at Buffalo, 2013). In females the stressors aren’t as debilitating cognitively as is shown in males; therefore there is a protective factor at work here. The common thread is estrogen; scientists should now be working on a way to get the protective effects of estrogen into males without having harmful side effects. With that technology both males and females will be on an even playing field in terms of stress.  

Danbolt, N. (2001). Glutamate as a neurotransmitter-an overview . Retrieved from http://neurotransporter.org/glutamate.html

Greenfieldboyce, N. (2011, December 08). Cagebreak! rats will work to free a trapped pal. Retrieved from http://www.npr.org/2011/12/09/143304206/cagebreak-rats-will-work-to-free-a-trapped-pal

Prefrontal cortex. 2013. In Merriam-Webster.com. Retrieved November 4, 2013, from http://www.merriam-webster.com/dictionary/prefrontal%20cortex

University at Buffalo, (2013, July 11). Females respond better to stress becasue of estrogen in the brain, animal study finds.. Retrieved from http://www.sciencedaily.com/releases/2013/07/130710061824.htm

Wei, J., & Yuen, E. (2013). Estrogen protects against the detrimental effects of repeated stress on glutanatergic transmission and cognition. Molecular Psychiatry, 1-11. doi: 10.1038 Retrieved from http://www.nature.com/mp/journal/vaop/ncurrent/full/mp201383a.html

 

Monday, October 21, 2013

NyQuil and its Effects on Paramecia


       One of my more interesting courses I'm enrolled in this semester is Genetics and I think most of the interest comes from the lab portion of the course curriculum. In particular the Paramecia lab that we just finished. The goal was to induce a mutation onto a culture of Paramecia but, the methods and materials were up to us to determine and experimentally figure out. My lab partner and I chose NyQuil cold and flu liquid to be our mutagen becuase my partner frequently uses NyQuil for her colds and flu symptoms so we wanted to see what it would do. We introduced the Nyquil to our culture of Paramecia at a variety of concentrations that were determined after a few test trials.

       Paramecia are generally studied for observation of their cilia on their outer membrane and their simple lifestyle. They are single-celled organisms that live in fresh water environments such as ponds (Genoscope, 2007). Paramecia are on average 120 micrometers in length and covered in hair-like projections called cilia, which are easy to observe under a light microscope in a laboratory (Fraga, 2001). The cilia on the Paramecia allow the organisms to swim in one direction, under normal conditions they swim only forwards but, when under toxic or abnormal conditions they reverse and swim backward to try to find a homeostatic environment (Fraga, 2001). When they are in a normal environment calcium channels are closed in the ciliated membrane so the concentration in higher outside the cell than inside the cell (Genoscope, 2007).When they are under abnormal conditions the calcium channels open and the calcium ions flush into the cell to reach an equilibrium between the outside environment and the inside environment (Genoscope, 2011). Once regular calcium levels are restored the calcium channels are closed again and Paramecia swim forward again.


Figure 2: Experimental wells
with NyQuil concentrations 
        The Paramecia culture was made with a wheat media and inoculated with the bacteria Klebsiella pneumoniae as the food source for the Paramecia. A flask containing 50mL of wheat media was inoculated with the bacteria and left to incubate for twenty-four hours, after the incubation period 1mL of Paramecia cells were added to the flask and left at room temperature to grow and multiply for a week. The prime time for experimentation on the Paramecia cells is 6-8 days old, this is when they are most healthy and abundant before they start to die off and become less viable. To keep a viable culture of Paramecia we remade cultures of cells each weeks for the following lab period.
Figure 2: Control Paramecia.
        Once the week was over the experimentation began. We did simple calculations to determine concentrations of NyQuil to Paramecia, starting with 50%, 10% and 5% solutions. After seeing that a 50% solution of Nyquil to Paramecia left the cells stunned and 100% dead we knew the concentrations we were looking for were much less than that.

       In the 10% solution the cells were still alive when transferred to a slide for observation. These cells had very different swimming movements, almost drunk-like. They were spinning and barrel-rolling around in the solution, after several minutes elapsed of the erratic swimming the cells started to slow down and eventually stop moving entirely. The cells seemed to have a slight change in shape and appearance as well as the abnormal swimming patterns, as seen in the photograph of the control cell to the cells in NyQuil solutions there are several morphological changes.

       These changes couldn't be defined as mutations in the actual genetic code or just situational mutations because the morphed cells didn't survive overnight to be transferred to a new culture medium. The 10% solution cells died rather quickly so we tried 5% which caused the same changes both behavior and morphological but again these cells didn't survive overnight. The next week we tried solutions with concentrations of 4%, 3%, 2%, and 1% NyQuil. With more time to keep these
cells in these concentrations we saw the cells start to appear to be exploding their membranes. In the picture the the left, you can see the bubbled out  of the membrane of the cells displaying the lysis process of cells. The behavior and morphological changes seen in the previous concentrations were also seen in these solutions but, with the decreased concentration of NyQuil the cells were swimming erratically for longer and longer periods of time before their swimming got to be sluggish.

        I think that if we had more time to experiment with these Paramecia we could have gotten a concentration more exact that would have sustained the life of the cells in the solution so we could have concluded that an actual mutation had taken place. Although I think we made some kind of change with the shape of the membrane of the cells as well as the swimming patterns there is no statistical data that we caused an actual mutation to the Paramecia cultures.


          References


Fraga, D. (2001, July 2). How ion channels control paramecia behavior. Retrieved from http://www3.wooster.edu/biology/Ciliates/citc/Para_behavior.html

Genoscope. (2007, Sept. 11). Paramecium teraurelia. Retrieved from http://www.cns.fr/spip/Paramecium-a-model-ciliate.htm


** All photos taken by myself (Abby Chauvin) in the lab using Image J software**

Monday, October 14, 2013

Caffeine, pick-me up or slow you down?

       Wake up, have a cup of coffee; that's the routine for many people including myself. As a young college student caffeine is vital in giving me the energy throughout the day to accomplish the long 'to do' list I always have. Caffeine is known, and widely consumed, for its characteristic of being a chemical stimulant and its ability to help resist sleep, keeping the consumer awake.
       Over the past thirty years the number of children and young adults drinking caffeinated beverages daily has increased by 70 percent (SNSF, 2013). This rise in the average is not forecasted to decrease any time soon because the market for energy drinks and caffeine-laden beverages keeps climbing and climbing every day (SNSF, 2013). There hasn't been much information on caffeine nor the effects it may or may not have on your body or brain. The average person doesn't know much about the way things work in their own body, and brain in particular, but researchers are working hard trying to crack the mystery of caffeine for everyone. The Swiss National Science Foundation recently supported a study on the effects of caffeine consumption on the brain development in children and young adults that was published in PLOS ONE.
        The brain is the most mysterious organ in the human body and it has taken years of research to know what we do know about the brain. Even with all that we do know, there are many vital defecits in the information known to us about the way the brain works in everyday life tasks. The brain controls everything that we do, think, feel, see, remember, say, hear, etc. The brain is shaped from our individual unique genes, but is largely influenced by individual environments and experiences of each individual person (Philips, 2006). No two brains are alike, they may have similar functions and connections but, they are completely unique from person to person. New brain cells are made by neurogenesis from stem cells (Medicine Net, 2012). Stem cells differentiate into brain cells or neurons, this process normally occurs in large bursts of growth reaching a macximum just before puberty, and again peaking during young adulthood. (Olini, Kurth & Huber, 2013) After the burst of growth comes a period of consolidation; where all the cells that aren't required are pruned, leaving only those needed (Olini, Kurth & Huber, 2013). This process of optimism of the brain cells is presumed to happen during deep sleep (SNSF, 2013). But, what if caffeine introduced to the young, still developing, brain affects that overall development process? That is the question the researchers wanted to test, although not with the human brain, they tested rats that were in the same developmental stage as children and young adults.
         The article The Effects of Caffeine on Sleep and Maturational Markers in the Rat, published in the journal PLOS ONE lays out the entire experimental procedure and complete results analysis. The rats in this study underwent surgery before the experiment to insert devices for longitudinal electrocortical readings after the caffeine was administered (Olini, Kurth & Huber, 2013). The rats were 28 male rats that were 22 days old, which is the pubescent age of these rats. In addition to the brain scan, the rats behaviors and anatomical development were also observed. The researchers took a base line development readings and observed control rats of the same age during the experimentation drinking only pure water.
       The results of the researchers study showed that those rats administered caffeine daily had far more neural connections in their brains than the control rats (Olini, Kurth & Huber, 2013). This indicated a slower maturation process of the rats' brain after caffeine, the behavior of these rats were also vastly different than those drinking water (SNSF, 2013). Rats generally become more curious as they age, but the caffeinated rats showed no curiosity, they remained timid and cautious compared to the control rats (SNSF, 2013). The brains of rats aren't an exact anatomical copy to the human brain, but there are similarities in how the connections are made in the brain during the maturation process of both rats and humans. These results cast a shadow over caffeine consumption in children and young adults; and if the average caffeine consumption in this age group doesn't show signs of reducing, there has to be something done about this problem or there could be severe developmental consequences. The brain is a mystery but science is getting pretty close to unveiling it.

References
 
Medicine Net. (2012, June 14). Definition of neurogenesis. Retrieved from http://www.medterms.com/script/main/art.asp?articlekey=18200
 
Philips, H. (2006, September 04). Introduction: the human brain. Retrieved from http://www.newscientist.com/article/dn9969-introduction-the-human-brain.html?page=1

Swiss National Science Foundation. (2013, September 24). Caffeine consumption slows brain development, rat study shows. ScienceDaily. Retrieved from http://www.sciencedaily.com/releases/2013/09/130924091323.htm#.UkbwFs34ItE.email

Olini, N., Kurth, S., & Huber, R. (2013). The effects of caffeine on sleep and maturational markers in the rat. PLoS ONE, 8(9), doi: 10.1371 Retrieced from http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0072539