Sometimes a new critical study is found which helps me and the other researchers understand a little better just how chondrocytes in cartilage turn into volumetrically growing bones and cartilage like growth plates. This is one of those studies. it may not seem like a lot for the person who just wants the step-by-step manual on how to grow taller, but for the researchers, this study is in my opinion one of the most insightful studies written due to the fact it shows just how the growth plates seem to be formed as a function of the overall volumetric shape or form that they are in.

Study #1: Growth Plate Chondrocytes Dedifferentiate in Monolayer but Redifferentiate in High-Density Pellet Culture, Sequentially Expressing Resting, Proliferative, and Hypertrophic Zone Markers

Center of Molecular Medicine, Department of Women and Child Health, Karolinska Institutet and University Hospital Stockholm, Sweden

Abstract

Longitudinal bone growth occurs at the growth plate, which is located near the ends of long bones. At the growth plate, cartilage expansion, through chondrocyte proliferation, hypertrophy and matrix production results in elongation of the bone. This process is orchestrated by the interaction of a large number of endocrine and paracrine factors. The study of these complex interactions has been limited by the lack of well characterized chondrocyte culture models and that growth plate chondrocytes cultured in mono-layer rapidly de-differentiate into fibroblast-like cells with high expression of Collagen type I (Col1) and minimal expression of chondrocytic markers, Collagen type II (Col2) and X (Col10). Dedifferentiated rat growth plate chondrocytes in monolayer were trypsinized and centrifuged to form high-density pellets. The cells rapidly (within 3 days) redifferentiated into Col2-expressing chondrocytes as evidenced by a more than 200-fold increase in Col2 expression, up to a level of expression similar to that of growth plate chondrocytesin vivo. We then assessed expression of newly identified resting (Sfrp5) and proliferative zone (Prelp) markers. With prolonged culture, these chondrocytes underwent a reproducible differentiation program characterized by an early rise in expression of resting zone marker Sfrp5, followed by a rise in the proliferation marker Prelp, and finally an increase in hypertrophic zone markers Indian hedgehog and Col10 expression. Our findings suggest that growth plate chondrocytes cultured in high-density pellets undergo a sequential differentiation program similar to growth plate chondrocytes in vivo. These findings may thus help to improve in vitro studies aimed at clarifying the interactions of endocrine and paracrine factors in the regulation of longitudinal bone growth at the growth plate.

Analysis

One of the things that keeps happening to us as any type of scientific researchers is that once we reach a certain level (usually extremely) of study, the understanding on how a phenomena of a physical mechanism actually happens is impossible. For the study of how the mechanism of growth plate process work at a microscopic level, it is the same as well. We just don’t know many things.

It does seem however that if chondrocyte cultures and growth plate chondrocytes are made into monolayers, they actually de-differentiate into a type of cell known as fibroblast-like cells with express only Collegen Type I, not the Type II and Type X which is seen in hyaline cartilage and hypertrophic epiphyseal zones.

The researchers either intentionally or accidently did something that resulted in the fibroblast-like cells to re-differentiate back into growth plate chondrocytes found in vivo. They ” trypsinized and centrifuged to form high-density pellets.” So it seems that they added some type of protein or enzyme called trypsin and took the monolayer cell cultures and spinned them until they turned into 3-D pellets that had the cells in a high density concentration.

The process took just 3 days for the fibroblasts-like cells to turn back into chondrocytes. This was checked with analysis of Collagen Type II testing, which showed a 200 times increase.

In addition, the researchers tested for certain “markers” which signify that a certain zone in growth plates were around

• Resting Zone Markers – Sfrp5
• Proliferation Zone Markers – Prelp
• Hypertrophy Zone Markers – indian hedgehog and Collagen Type X

They state, “Our findings suggest that growth plate chondrocytes cultured in high-density pellets undergo a sequential differentiation program similar to growth plate chondrocytes in vivo.”

This shows an important point which we will need to understand if we ever plan to create a type of research program or do any type of laboratory experiments to get a type of cartilage tissue that functions almost exactly like the growth plates found in human bodies.

We need to form 3-D pellets for growth plates to work, not monolayers. Any type of cartilage structure we decide to create must have a 3-D form, not something very thin. 

Pellets with the diameter large enough to be re-implanted into lab rabbits to test to see if they restart the longitudinal growth process will be done first. That was proven to be successful in studies done by a group of researchers in Hong Kond.

Eventually one day that pellet must be big enough in width, and have the right thickness as well as structure strength to be implanted back into an adult human’s tibia or femur to get them to start growing their bone longer again.

Sometimes someone comes along and shows me a study that pushes the endeavor a little further and that is the type of help I hope we as the researchers can get from the readers.

For most of the stuff that the readers show me or direct me to, I look at the material to see if there is anything useful that I and other researchers can use. I am very grateful to the person who showed me the study “Engineering growing tissues” by Eben Alsberg. From just a quick Google search on Dr. Alsberg, it seems that he is cased in the Biomedical Engineering Department at Case Western University, which I remember had a very strong engineering school. I would look through his profile and research to see what other papers he has written about tissue engineering over the years. There are some interesting papers he has been a coauthor to, including…

1. “Spatiotemporal regulation of chondrogenic differentiation with controlled delivery of TGF-β1 from gelatin microspheres in mesenchymal stem cell aggregates”
2. “Stromal-cell-derived factor (SDF) 1-alpha in combination with BMP-2 and TGF-β1 induces site-directed cell homing and osteogenic and chondrogenic differentiation for tissue engineering without the requirement for cell seeding”
3. “Engineered cartilage via self-assembled hMSC sheets with incorporated biodegradable gelatin microspheres releasing chondrogenic growth factor”
4. “Controlled and sustained gene delivery from injectable, porous PLGA scaffolds”
5. “Chondrogenic differentiation of human mesenchymal stem cell aggregates via controlled release of TGF-b1 from incorporated polymer microspheres.”

These studies are the most interesting to me but the last is is particularly attractive to read through. I guess I will get to it in another day.

It is interesting that he does a lot of research in the biomedical application of electrospinning, since I have stated that electrospinning is one of the more likely ways that human cartilage can be grown rather easily for growth plate re-implantation. I wrote a post about this idea entitled “Increase Height And Grow Taller Through Bioprinting And Electrospinning“. At this point, I will move away from Dr. Alsberg research and look at what other university tissue engineering and biomedical engineer professors have been doing research on.

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The 2nd study that he would refer me to showed that there has been even more ways and methods that researchers have done in recent years that allowed for growth plate regeneration.

IGF-I RELEASING PLGA SCAFFOLDS FOR GROWTH PLATE REGENERATION

Sharath Kumar Chinnakavanam Sundararaj – Master’s Theses – May, 5th 2010 – UKnowledge
University of Kentucky, sharath.sundararaj@uky.edu

Abstract

Growth plate is a highly organized cartilaginous tissue found at the end of long bones and is responsible for longitudinal growth of the bones. Growth plate fracture leads to retarded growth and unequal limb length, which might have a lifelong effect on a person’s physical stature. This research is a tissue engineering approach for the treatment of growth plate injury. Insulin-like growth factor I (IGF-I), which can stimulate cartilage formation, was encapsulated within PLGA microspheres that were then used to form porous scaffolds. The release profile of the IGF-I from the PLGA scaffold showed a biphasic release pattern. In vitro studies were done by seeding rat bone marrow cells (BMCs) on the top of IGF-I encapsulated PLGA scaffolds, and the results showed an increase in cell multiplication and glycosaminoglycan content. The final in vivo studies were conducted by creating growth plate injury and implanting scaffolds in the tibiae of the New-Zealand white rabbits. Histological analysis of tissue sections showed regeneration of cartilage, albeit with disorganized structure, at the site of implantation of IGF-I encapsulated scaffolds. This work will be a significant step towards tissue engineering of growth plate cartilage.

KEYWORDS: PLGA, Tissue engineering, growth plate, drug delivery and IGF-I

Analysis #1: I have reviewed multiple Ph. D theses before and it was very mentally tiring work to read through what are usually 200+ pages of scientific data and charts. This Master’s Theses maybe only 70 pages but it still will take a lot of work to get through.

The overall theses is not to actually grow a growth plate from scratch using a seed from an autologous explant, but it is more towards trying to grow a similar physeal cartilage like the growth plate in the fractured area when the growth plate experiences a type of injury that might cause something like a bony bridge.

1. IGF-1 was somehow put inside/encapsulated in microspheres that are made out of PLGA.
2. The microspheres were used to make into porous scaffolds.
3. The IGF-1 will eventually get released from the PLGA microspheres as either the microspheres and/or the scaffold starts to disintegrate
4. The way that the IGF-1 is released is shown to have two phases.

The studies to look at the release profile and release characteristics of the IGF-1 was done AT FIRST in in-vitro environment.

1. Some rat marrow cells were taken out from the actual lab animal (probably using a similar invasive method as a biopsy)
2. The cells were put on top of the scaffold which already has the IGF-1 inside.
3. The result is that there was an increase in glycosaminoglycan content and cell multiplication

The studies were then done AT THE END in vivo

1. Some lab rabbits, of the New Zealand type was taken. 
2. The growth plate in their tibia was broken to create an injury, which left a defect or distracted area where intact, continuous growth plates used to be.
3. The scaffold with the IGF-1 inside was put in the area of empty space left by the growth plate
4. Histological analysis (which is just taking a microscope and looking at the tissue being analyzed to see the cell structure, cell number) of tissue sections showed regeneration of cartilage.
5. The growth plate cartilage that was created however showed a disorganized structure

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What is PLGA? – From Wikipedia – poly(lactic-co-glycolic acid) is a copolymer which is used in a host of Food and Drug Administration (FDA) approved therapeutic devices, owing to its biodegradability and biocompatibility

What is a Copolymer? – From Wikipedia – A heteropolymer or copolymer is a polymer derived from two (or more) monomeric species, as opposed to a homopolymer where only one monomer is used.

What is a Polymer? – From Wikipedia – is a large molecule composed of many subunits, known as monomers…  Polymers range from familiar synthetic plastics such as polystyrene (of styrofoam) to natural biopolymerssuch as DNA and proteins that are fundamental to biological structure and function. Polymers, both natural and synthetic, are created via polymerization of many monomers

What is a Monomer? – From Wikipedia – refers to the organic molecules which form synthetic polymers, such as, for example, vinyl chloride, which is used to produce the polymerpolyvinyl chloride (PVC).

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 So what was the overall message of the Thesis?

(Page 2) Tissue engineering of cartilage is gaining heavy importance in the field of tissue engineering. But most of the research till now is mainly focused on the regeneration of articular cartilage and very little progress has been made in tissue engineering of growth plate. This work was aimed at providing a significant contribution towards tissue engineering of growth plate.

We hypothesized that microencapsulation of IGF-I in PLGA would be an appropriate growth factor delivery system for regeneration of growth plate cartilage both in vitro and in vivo.

THe 5th section of this Master’s Theses is on the Discussion. It is broken up into certain sections

• Release profile of protein encapsulated scaffolds degraded in PBS 
• Release profile of protein adsorbed and protein encapsulated scaffolds
• Release profile of protein-encapsulated scaffolds degraded in cell culture medium 
• Release profile of protein-encapsulated scaffolds degraded in cell culture medium 
• along with macrophages

2nd part: Degradation of PLGA scaffolds

• Mass loss of scaffolds degraded in PBS 
• Mass loss of scaffolds degraded in cell culture medium

3rd part: In vitro cell studies

• Seeding the cells with fibrin glue 
• Confocal imaging of BMC-seeded scaffolds
• Analysis of GAG content from the BMCs

4th part: In vivo studies 

• Scaffold used in the animal surgery
• Radiographic images
• Histology

Last section #6: CONCLUSION 

The main aim of this research was to develop a biodegradable drug delivery device for the treatment of the growth plate injury. The regeneration of the growth plate cartilage in the site of injury has confirmed that the IGF-I was successfully released from the porous PLGA scaffold and induced the formation of chondrocytes and extracellular matrix. The release studies showed that microencapsulation of IGF-I was a suitable method for storage of the drug within the scaffold that is going to be released. The release studies and the in vitro cell studies confirmed that the device was capable of releasing the drug in a required pattern and bioactive state. The IGF-I encapsulated porous PLGA scaffold implanted in the rabbit model for treatment of growth plate injury has successfully regenerated cartilage at the site of injury, even though it was not effective in correcting the defect in the bone angle that was caused due to the injury. The results of this research has shown that using porous PLGA scaffolds with IGF-I encapsulated in the form of microspheres will serve as an appropriate drug delivery system for the cartilage regeneration. This work will be a significant step forward in the tissue engineering of growth plate cartilage.

Analysis #2

It seems that as long as there is some growth plate left in the animal, regeneration is possible using just a scaffold of the right type (this time the right material is PLGA) and some IGF-1. The IGF-1 did result in cartilage being regenerated but it did not manage to fix the defect in the bone angle that was originally induced to create the bone injury.

Me and the other researchers have found this type of thing before, and this theses shows that if some growth plates is missing, we can recreated it. The histological analysis seems to show that the chondrocytes that were in the newly created cartilage were not organized or columnar stacked like how they should be to have the hypertrophic zone which would lead to maximum longitudinal growth rates.

This suggests that while IGF-1 is affective alone in a scaffold, there might be some growth factors which might be better to towards forming chondrocytes that are more organized and in the right position for better growth chances.

Something to realize is that if there is NO growth plate cartilage left, then this method would not work There has to be at least some cartilage left. This method is similar to what we see in techniques which result in articular cartilage layer thickening where some articular cartilage is regenerated on the top surface to decrease osteoarthritis and cartilage degeneration associated with old age.

The problem is that if we are talking about adult human with absolutely no cartilage to start with, then wee need more than just a scaffold and a growth factor as a stimuli.

This study/theses was useful and will be used in future posts to show that it is possible to heal over defects in growth plate cartilage with new cartilage. However it is not enough for people with absolutely no physeal cartilage to start with.

I was planning on going through a dozen websites that dealt with height increase and how to grow taller. These were old websites and urls I had found many months ago, around August of last year which were around and that I found which I had always wanted to go back over to see if there might be something of real value that a researcher can glean from them.

1. Height Discussion – URL: www.heightdiscussion.com – Tagline: Grow Taller Community
2. The Centre Of Anthropometrical (Orthopaedical) Cosmetology And Correction – URL: www.rucosm.com – Tagline: expert in legs correction & lengthening since 1992
3. Is There A Way To Get Taller – URL: isthereawaytogettall.blogspot.com – Tagline:
4. Yogasagar Yoga Therapy – Increase Height – URL: www.yogasagar.org/yoga-therapy/increase-height/ – Tagline:
5. Exercises To Increase Height – URL: www.exercisestoincreaseheight.org/ – Tagline:
6. LSJL Tester – WordPress – URL: lsjltester.wordpress.com/ – Tagline: This WordPress.com site is the cat’s pajamas
7. Short No More – www.shortnomore.com/tag/deep-sleep/ – Tagline:
8. Charged Audio – ChargedAudio™ Grow Taller Now NLP Subliminal CD (D19) – URL: www.chargedaudio.com/bodyfitnessseries/d19.html – Tagline: Grow Taller. It’s No Accident. In 14 Days, You Can Learn How!
9. How To Grow Taller Answers – URL: howtogrowtalleranswers.com/how-can-i-open-my-growth-plates-and-grow-taller.html – Tagline: Grow Taller Answers, Advice And Information
10. Helpo Growth Formula – URL: helpo-grow-taller-supplements.blogspot.com/ – Tagline: Tips to help you grow taller
11. Height-Increasing – URL: www.height-increasing.com/category/facts-growth-height/ – Tagline: Your Complete Guide For Height Increasing
12. How To Get Taller – URL: howtogettaller.org/the-role-of-growth-hormone-for-height/ – Tagline: Be Tall and Be Cool

Analysis

The website Height Discussion turns out to be a very new forum which has a few posts and threads. There are a few moderators but it does not seem that this new grow taller forum is going to get enough traction to attract the high number of people needed. There seems to be already a few threads started that are spam like in nature so I would avoid this website/forum for now until they can get the spam issue in check.

I had originally put this website called The Centre Of Anthropometrical (Orthopaedical) Cosmetology And Correction into the Resources section as a link to limb lengthening services offered in a part of Russia. The website does look legitimate and linked to some medical clinic in Volgograd, Russia. The contact us address is listed in Volgograd so this center does seem to exist and offer the limb lengthening servies. They are associated with the Ilizarov Clinic in Kurgan.

The blog called Is There A Way To Get Taller based off of the blogspot platform is now gone. If you put the domain name in, nothing shows up.

The website section on Yogasagar Yoga Therapy for learning how to increase height has also been deleted. It seems that the entire website for yogasagar has been deleted as well. There is just a page with links to yoga articles which is very generic and many ads around. Nothing useful

The website Exercises To Increase Height seems to have also been shut down and the website completely removed. There is nothing there at all.

The blog LSJL Tester seems to be a valid blog which wanted to test the idea that Tyler proposed but it seems that it has not been updated on in quite a while. It was started last year (2012) around the May-April time where the person was doing a good job in documenting the progress. However it seems that no progress had been made and nothing has been written about. The person did disappear for a full year and say that they came back but it seems that disappeared again. Overall, this website also fails in its ability in the person to keep writing posts to inform the readers what is happening.

The website Short No More seems to have also disappeared. If the person who created it thought that the process in doing the research to find a way to “cure” shortness was going to be easy, then they made a big mistake. I checked both the domain names ShortNoMore.com and ShortNoMore.org and found nothing.

The Charged Audio website seems to be selling an Audio product called Grow Taller Now NLP Subliminal CDs which are based on the idea of NLP and subliminal messages reprogramming the subconscious mind. It seems that the website for Charged Audio sells all kinds of CD programs that claim to be able to help a person grow taller using hypnosis and binaural beats technology. I wrote a an old post about the possibility of using Binaural Beats to increase height “Increase Height And Grow Taller Using Brain Wave Binaural Beats“ where I said at the end that the product will not work.

The website How To Grow Taller Answers seems to be another one of those websites created to develop a web of backlinks to help certain websites move up in Google Rankings. The website seems to be just 4 pages long, with Google Adsense ads everywhere, and ads selling the Grow Taller 4 Idiots and Grow Taller 4 Dummies E-Products, which obviously don’t work. There is a section as the bottom which does list soem good height increase resources but everything else is just a long sales page that probably was created 4-5 years ago to sell the scam E-Products.

The blog Helpo Grow Taller Supplements was created to sell the product Helpo Growth Formula.  I wrote a review on the Helpo product many months ago in the post “Product Review VIII: Helpo Growth Formula”. In that review I got slightly angry in bashing the product and whoever sold it to scam people out of their money. This blog is useless as a resource.

The website Height-Increasing seems to be a 2 page website created to give only very superficial information about common sense ideas on how to either eat healthy food or look taller from changing one’s clothing and fashion. It was not helpful at all. Created back in 2006, the website has not been updated or added upon since 2009. The number of posts is only around 5-6, but the articles are reasonably informative, which is something one can find in any Grow Taller E- Book.

The website How To Get Taller (dot org) is another website created by someone back in 2009 with two posts on the wordpress and left to die out. Nothing has been added upon in years.

Analysis On Additional Websites…

Since this post is to do a batch like process of going through and reviewing as many other height increase websites and blogs as possible, I wanted to add a few more websites to the group for a 2nd round of analysis.

1. Growing Taller Stretches – URL: growingtallerstretches.com/ – Tagline: Growing Taller Stretches
2. How to Grow Taller Naturally and Increase Height – URL: www.growtall.asia – Tagline: Increase stature and grow taller tips, growing taller secrets, grow taller stretches and grow taller exercises
3. All Height – URL: allheight.blogspot.com/ – All about height. – Tagline: Serving detailed height measurement information.
4. Grow Taller Secret – URL: grow-taller-secret.blogspot.com/ – Tagline: Free tips on How to Increase your Height
5. Grow Taller Naturally – URL: www.coprobol.org/ – Tagline: Easy techniques to help you grow taller.
6. Hypnosis 2 Height – URL: www.hypnosis2height.com/ – Tagline: Complete Step-by-Step Guide to Grow Taller with Hypnosis

Full Analysis 

The website Growing Taller Stretches seems to be another sales pages created with SEO in mind. There is only 4 posts ever written with Google Adsense strategically placed selling the Grow Taller 4 Idiots product. The website is not helpful as a resource.

The website Grow Tall with a dot asia ending is one of those websites which turns out to be much more useful and informative than 95% of the grow taller websites I find out there. It seems that the person who created the website back in 2008 really was trying to do some research to see if anything would work. He/she wrote a few posts which are similar in subject as what are found in this website, looking at how the element of stature/height affects society and people’s perception of what it means to be tall or short. From April of 2008 until 2010, the creator did put up around 60-100 posts which are very short in length and have useful information.

Something I would want to criticize the website designer is the fact that the way they put Adsense Ads so blatantly on the webpage and keep trying to push certain supplements which they say can help people grow taller by using Amazon Affiliates. They wrote very short posts about reviewing Robert Brand’s books and Peak Height but they still are promoting those produts when it should not help.

In the end, I am not sure whether this Grow Tall website was created just to make money through ads and affiliate marketing, or was it created to maybe help people. By judging the website for the quality of the posts it writes, I would say that the website was made more likely to get traffic, and eventually sell products which would never work.

The website All Height which is based on the blogspot platform seems to be created by a filipino and used to document the heights of filipino celebrities and a few other foreign celebrities and famous people. the person started the website around the same time that I did and the amount of content they have written is quite high so this website is a very good resource for a person who might be interested in knowing how tall a filipino celebrity is.

The website Grow-Taller-Secret which is also created using the blogspot platform from 2006 to 2009 has been one of the only websites that I have seen which has actually some really good, useful, in depth content. This is a website I did put in the Resources page.

The website CoProBol.com turns out to be just another website that is used as a sales page linked to the E-Product Smaller To Taller, which a regular reader of this website has given me.

The website Hypnosis 2 Height which is called The Step-By-Step Guide To Grow Taller With Hypnosis is one of those websites which is actually kind of informative and useful if a person wanted to try the ida of using hypnosis to increase their height. The website is only 9 pages large with a small section for each part, but I would say that the information is still kind of good, although not a lot in quantity. It was referenced by Giant Scientific in the resources section for years so it is somewhat credible.

Conclusion: 

These 18 websites I have found referenced around the forums and other websites on the website are mostly useless, sales pages selling E-Books but a small percentage have been helpful. A lot of the websites are now gone and some of them will probably also disappear in the years to come.

Update #3: What Has Changed Over The Last Month Towards Me Getting A Taller, Slimmer Body – June 1st, 2013

It is already June, and this website/blog has been around almost a year. The research has been wonderful and in this month actually there was a few big breakthroughs. I’ve found evidence that growth plate regeneration has already been done by a few biomedical researchers in university laboratories in the USA so now the big thing to figure out is what is the least invasive approach to embed the growth plates into the adult human bone without causing too much pain and decrease the amount of time needed to recover.

So what exactly has been going on in my life? Have I got any taller from May to June?

Diet & Weight: I have recently dropped in weight from decreasing my food intake over the last month. These are the things I have done to loss weight.

• I managed to decrease my soda/pop addiction to a fraction. Where it used to be I would drink Sprite & Cola every other day, I am now drinking 2 liter bottle of water each day for the Summer heat. 
• Since moving back to Seoul I had to give away the bike and put that in storage.
• My exercise program now focuses on doing pushups using Push Up Handles, Jump Rope for the aerobic workout, and sit-ups to decrease the fat content in my stomach.
• I’m supposed to do 40 pushups, jump rope 800 times, and do 60 situps today. That is not a lot compared to most guys who work out but I just got back to exercising after not doing it for over a year.

I don’t have a weight scale around the apartment so I am just guessing right now. I have noticed that right after I wake up, my stomach seems to be smaller and thinner. Current Weight: about 209 lbs.

Height Increase: There are no swimming pools in Seoul that are cheap and convenient so one of the only exercises I really like is not available. The good one I have found is in Everland but there is no way I am going to get on a bus for 2-3 hours just to use a pool.

So I stick to stretching. The main stretches I do, which I do 3 times a week, usually around 10 PM is with a yoga mat…

• Yoga – The local E-Mart sell perfectly reasonable priced yoga mats for 12,000 won and that is what I use to do the stretching. I focus on 3 main stretches, which is to… 1.Twist the torso, 2.Pull down on the torso, 3.Pull up on the torso.

Height Gain From May: 

I did not see any increase in height this month. My hair has grown out so the measurement error has gone back up so any slight increase I notice from the morning measurement I don’t trust.

If you want, you can read the update I did in May in the post “Update #2: What Have I Been Doing To Grow Taller And Become Healthier – May 1st, 2013”