[Note: This is one of the most important posts and summarizes the research that has been going on for the last 4 months.]

Me: I think it is time to show to the world just what the last 4 months of research has brought up. Here is my claim from the PubMed studies I’ve found…. “Epiphyseal growth plate regeneration, regrowth, implantation, and transplantation is completely possible!”

We have already shown that we can create chondrocyte mediums who can allow us to grow chondrocytes into cartilage.  We can even buy the chondrocyte mediums with the progenitor cells from the companies I have already listed on this website. Here are the 3 posts I’ve written before that I want to cite.

From the last post, “Epiphyseal Plate Transplantation Through Vascularization (Breakthrough!)” we have seen that growth plates can be transplanted into new subjects and the subject will go through longitudinal growth.

Let me cite previous posts looking at this subject.

1. Hyaline Cartilage Engineered By Chondrocytes In Pellet Culture Compared To Cartilage Implants (Important)
2. Injecting Chondrocytes Into The Physis From Physeal Distraction Of The Epiphysis
3. Increase Height Through Surgical Method By Cartilage Harvesting And Chondrocyte Implantation With Growth Factor Injections

The first post showed that growth plates were being grown from chondrocytes taken from the epiphyseal plates of the host or subject and then

The 2nd post showed that if you inject chondrocytes into the fracture of a distraction where there is already the growth plate it will lead to increased longitudinal growth.

The third post was my own first original idea on how the process of long bone distraction, cartilage harvesting, and then re-implanting of chondrocytes would work.

The 6 steps for the proposed idea were…

1. Remove progrenitor mesenchymal stem cells or chondrocytes from the subject.
2. Grow the MSCs & chondrocyctes until they form the rest zone formation of growth plates.
3. Implant this into a distraction of the long bone. 
4. If not in-vitro grown epiphyseal plates, transplant it.
5. Get the host body to accept it. 
6. Get longitudinal growth to start again 

At this point I can say with complete confidence that every single step in my proposed method to increase height, at least one study or experiment has already been done and been proven by researchers around the world. This is the real thing, and every single step has been proved successful in implementation.

Me: This story was also found from the Make Me Taller Forum (source link) and from the first look of it, it doesn’t seem to be at least an April Fool’s joke like the last one.

Analysis: The thing is that while the article says Pierre grow 2 inches from 5′ 7″ to 5′ 9″ but in the video he actually claims only 1.5″ of height increase, which for the height increase seeker’s community is a big difference. We are talking about at least a 1 cm difference in claim between the two sources. In the Mirror UK it seems he increased his height by 3.15 cm. It also shows that he is not 5′ 9″ but “just shy of it”. He claims that if you join in the 6 weeks program you are gauranteed to increase in height by 2 cm. For Pierre he has been doing those exercises for “a couple of years”. It involves combining pilates and yoga along with strength work and speed work. There is a device which is similar to the traction machine which stretches out the body (at least the vertebrate). In terms of the science behind the idea, there is some very similar ideas to Sky’s Old theory. Pierre seems to suggest that high intensity exercises like sprints help elevate the HGH release rate which we researched and shown to be real. In addition those sprints will cause microfractures which will eventually heal and lead to bone lengthening (or at least he claims). There is also height increase “by inducing spinal decompression which stimulates growth by increasing the fluid capacity in the spinal disks” as Pierre says. The GymBox apparently is willing to give you a money back refund if you try out the 6 week program and don’t grow the 2 cm that they promise you. So the program seems to involve the science of spinal decompression, inducing microfractures, and increased HGH levels. When I looked at this Pierre guy on Google, he turns up to be a real trainer based on the UK with his own website, company, and LinkedIn profile.

Conclusion: From a quick google search on the term “A-Grow-Bics” there are a few articles all written by UK sources looking at this claim. There was an article written by Daily Mail UK, Standard UK, Reveal UK, Mirror UK, Female First UK, and also the GymBox website. The gym is held in London and you can join the 6 week program for 200 pounds and they offer a refund if you don’t get the 2 cm increase that they promise. Tyler on one of his posts years ago said that he hoped that one day people can go into the local gym and ask a personal trainer to get them on a program which would allow them to increase height. Maybe that day is finally here.

You can watch the video below where the fitness trainer Pierre Pozzuto claim a few things which seems very incredible.

Daily Mail UK article link HERE…

New gym class A-Grow-Bics promises to add inches to your height – but is it a tall story?

By LUCY WATERLOW

PUBLISHED: 11:51 GMT, 22 October 2012 | UPDATED: 12:17 GMT, 22 October 2012

It sounds ideal for short men like Simon Cowell and Jamie Cullum who may want to be taller than the ladies in their lives, while petite women such as Kylie Minogue and Lea Michele could ditch their sky-high heels if claims from a new gym class are true.

Fitness chain Gymbox say they can have developed a workout that guarantees to make you taller. They are so confident of their claim, they even offer money back to anyone who tries it and doesn’t grow at least 2 cm.

Fitness trainer Pierre Pozzuto developed the gym class which involves a mix of pilates, yoga, strengthening and speed work – plus a session on a scary-looking 15th century-inspired rack.

Such implements were once used to torture people but Pierre insists it isn’t a painful experience as it’s designed to gently stretch the body once the muscles are warmed up.

Pierre developed the six week course because he was keen to increase his own height from 5ft 7.

He said: I wasn’t the biggest guy in the world and it always bothered me, so at the age of 26, I decided to do something about it. I set about designing a set of exercises to help me grow and they worked – so much so that my height jumped to nearly 5ft 9 in only a few months.

Tall order: From left, could Simon Cowell, Jamie Cullum and Tom Cruise gain in stature by following fitness trainer Pierre Pozzuto’s techniques?

‘When I told a few people, they begged me to help them grow too and that’s the aim with the A-Grow-Bics class.’

Pierre said his class works by inducing the Human Growth Hormone (HGH) through repeated sprint exercises.

He said repeat sprinting also ’causes micro fractures in the bones that repair in a week encouraging them to strengthen, remould and grow.’

The rack, yoga positions and inversion exercises, such as hanging upside down from the feet, are also said to increase height ‘by inducing spinal decompression which stimulates growth by increasing the fluid capacity in the spinal disks’, according to Pierre.

But Dr James Noake, an expert in sport and exercise medicine, told Style magazine he was skeptical about Pierre’s claims.

‘The discs between our vertebrae are made of tough material, with little capacity for expansion, and the rigid join structures either side of them will be resistant to lengthening,’ he said.

He added that the rack was ‘highly unlikely’ to permanently help someone become taller while encouraging micro fractures was ‘a little alarming’ and ‘unlikely’ to be caused by speed work in those with healthy bones.

But Gymbox insist they can add inches to anyone who follows Pierre’s method.

Managing director Richard Hilton said: ‘People join our gyms for all sorts of reasons, but one of the more unusual requests our trainers often receive is for exercises to aid growth. While we know this is a tall order, Pierre’s techniques are head and shoulders above the rest and really do work.

‘We are so confident in this class that we guarantee all participants will grow by at least two centimetres – if not, we’ll happily give them their money back.’

The six week A-Grow-Bics is held at Gymbox’s Farringdon, London, branch, at a cost of £200.

Watch Pierre Pozzuto explain his height-increasing techniques below…

When I was going through the Make Me Taller Forums today I found a link that someone posted about another possible way to increase height which seems to have been developed by a Israeli academic Professor Ura Schmuck and was being tested at some Swiss Laboratory. (source link)

The big thing to note is that this story was written by the sensations tabloid “The Sun” whose job is to write up exaggerated, and maybe even sometimes fake, stories which will get eyeballs and ratings and press. I am not sure if the fact that the former french president Sarkozy did use the technology or not was true.

Analysis: My issue with this story is that when I tried to find out more about this “Stature Augmentation Treatment” and “Ura Schmuck” nothing came up. Then I sort of realize the name and the date.

Conclusion: The fact is that the Doctor’s name is “Ura Schmuck” is a homonym for “You Are A Schmuck” meaning that if you really believed in this article, you are an idiot, and that the writer wrote this article on April 1st means that they were just playing an April Fool’s joke on the readers. Get That Part? However, I stated that I would look at every single idea ever posed and this article was still fun to write up.

The full article is found from HERE and is posted below…

Me: This was a patent I found. In the patent however the growth modulator is used to compensate for the disproportionate growth found in defective vertebrate. The growth modulator is generally administered using a delivery device suitable for delivery of therapeutics into bone, such as a suitable needle for delivery of liquid or semi-liquid (e.g. semi-viscous liquid) formulations, or via a catheter device for delivery of solid and semi-solid formulations. It seems that the long metal needle used to be stuck into the vertebrate growth plate is a physiologically acceptable carrier for administration to an epiphyseal growth plate. From the patent…

As typically observed in congenital scoliosis, abnormal vertebrae growth arises from the premature partial fusion of the epiphyseal growth plate. In the region of premature partial fusion, the chondrocytes generally exhibit decreased activity, thereby retarding the growth of the defective vertebrae in the region of the partial fusion. The end effect is concavity in the region of partial fusion

From the patent…“The intervertebral disk 134 can also serve as a temporary reservoir of growth modulator. In this delivery methodology, termed discal delivery and shown in FIG. 12, the growth modulator is deposited within the fibrocartilaginous structure of the disk 134 wherefrom it diffuses into the adjacent vertebrae. To promote diffusion into the appropriate target area, the growth modulator is generally deposited into the disk in the area immediately adjacent to the area of the vertebrae 120 requiring growth modulation.”

Analysis: 

There is two major proof of concept from this patent.

1. The idea that one can just directly inject growth modulators into the epiphyseal plates, whether they are from either the irregular bones of the vertebrate or the long bones in the legs/arms to make the cartilage plates grow faster and further
2. The growth modulator seems to be okay to be used for even partially fused epiphyseal plates, which seems to prove the idea Tyler has been proposed that even partially fused epiphyseal plates can overcome a bone bridge and cause longitudinal growth.

The  positive growth modulator comprises one of…

1. a natural growth hormone, somatotropin
2. an exogenous bone growth stimulant
3. a somatotropin analogues
4. agent comprises fibroblast growth factor (FGF), bone morphogenic proteins (BMPs), stem cells, vascular endothelial growth factor (VEGF), angiogenic growth factors, blood flow control agents and vascularity enhancing factors.

The growth modulator has 3 key components.

1. Stem cells
2. A chondrocyte growth factor
3. A natriuretic peptide

Patent application title: MODULATING BONE GROWTH IN TREATING SCOLIOSIS
Inventors:  Kieran Murphy (Toronto, CA)
IPC8 Class: AA61K3512FI
USPC Class: 424 937
Class name: Drug, bio-affecting and body treating compositions whole live micro-organism, cell, or virus containing animal or plant cell
Publication date: 2010-09-02
Patent application number: 20100221229

From source link HERE…

Abstract:

The present specification provides, amongst other things, a method of treating scoliosis comprising delivering a therapeutically acceptable amount of a growth modulator to an epiphyseal growth plate to correct or compensate for disproportionate growth.

FIELD

[0002] – The present invention pertains to the field of scoliosis, in particular to the treatment of scoliosis using growth modulators.

SUMMARY

[0003] – According to a first aspect, provided is a method of treating scoliosis, comprising delivering a therapeutically acceptable amount of a growth modulator to an epiphyseal growth plate to correct or compensate for disproportionate growth.

[0004] – According to a further aspect, provided is a composition comprising a therapeutically acceptable amount of a growth modulator and a physiologically acceptable carrier for administration to an epiphyseal growth plate. According to another aspect, provided is a method of treating scoliosis, comprising the delivery of a growth modulator to a target region of an epiphyseal growth plate for altering the growth in said target region.

[0005]According to a further aspect, provided is a method of treating scoliosis, comprising delivering a therapeutically acceptable amount of a growth modulator to selected portions of an epiphyseal growth plate to correct or compensate for disproportionate growth.

Claims:

1. A method of treating scoliosis, comprising delivering a therapeutically acceptable amount of a growth modulator to an epiphyseal growth plate to correct or compensate for disproportionate growth.

2. The method of claim 1 wherein said growth modulator is a positive growth modulator to stimulate growth.

3. The method of claim 2 wherein said positive growth modulator is introduced into a vertebrae adjacent to a defective vertebrae in a manner that physiologically compensates for the disproportionate growth of said defective vertebrae.

4. The method of claim 3 wherein said positive growth modulator is introduced into an epiphyseal growth plate of said adjacent vertebrae in a region adjacent to a partially fused epiphyseal growth plate of said defective vertebrae.

5. The method of claim 2 wherein said positive growth modulator comprises one of a natural growth hormone, somatotropin, an exogenous bone growth stimulant, and a somatotropin analogues.

6. The method of claim 2 wherein said positive growth modulator comprises a biosynthetic human growth hormone

7. The method of claim 6 wherein said biosynthetic human growth hormone comprises one of Nutropin (Genentech), Humatrope (Lilly), Genotropin (Pfizer), Norditropin (Novo), and Saizen (Merck Serono).

8. The method of claim 2 wherein a positive growth modulator comprises a suitable inhibitor of endochondral ossification.

9. The method of claim 8 wherein said inhibitor of endochondral ossification comprises chondromodulin-1.

10. The method of claim 2 wherein said positive growth modulator comprises one of stem cells, a chondrocyte growth factor (CGF), and a natriuretic peptide.

11. The method of claim 1 wherein said growth modulator is a negative growth to inhibit growth.

12. The method of claim 3 wherein the negative growth modulator is introduced into an epiphyseal growth plate in order to decrease an extent of wedge formation in a growing defective vertebrae.

13. The method of claim 11 wherein said negative growth modulator comprises an agent that ceases chondrocyte activity or promotes ossification of the cartilagenous tissue.

14. The method of claim 11 wherein said negative growth modulator comprises agents that increase vascularization or promote the ossification of the epiphyseal growth plate tissue.

15. The method of claim 11 wherein said agent comprises fibroblast growth factor (FGF), bone morphogenic proteins (BMPs), stem cells, vascular endothelial growth factor (VEGF), angiogenic growth factors, blood flow control agents and vascularity enhancing factors.

16. The method of claim 11 wherein said negative growth modulator comprises a chemotherapeutic agent therapeutically effective to interfere with an ability of cells to grow.

17. The method of claim 16 wherein said chemotherapeutic agent comprises one of alkylating agents, anthracyclines, cytoskeletal disruptors, epothilones, inhibitors of topoisomerase II, nucleotide analogs and precursor analogs, Peptide antibiotics, retinoids, and vinca alkaloids and derivatives.

18. The method of claim 11 wherein said negative growth modulator comprises one of an antibiotic, an antimitotic, an antimicrobial, an alcohol, a protein, a disinfectant, a hydroxyapatite, and a metal.

19. A composition comprising a therapeutically acceptable amount of a growth modulator and a physiologically acceptable carrier for administration to an epiphyseal growth plate.

20. A method of treating scoliosis, comprising the delivery of a growth modulator to a target region of an epiphyseal growth plate for altering the growth in said target region.

21. A method of treating scoliosis, comprising delivering a therapeutically acceptable amount of a growth modulator to selected portions of an epiphyseal growth plate to correct or compensate for disproportionate growth.

Me: This is going to be the start of a proposed idea on how to increase height using a two step process. From a lot of the research, we have already found many, many ways to make bone fractures heal. That part is easy. In addition, we have also found many ways to induce the creation of chondrocytes from some form of external stimuli, whether chemical, mechanical, electrical, to cause the right type of differentiation in progenitor cells. Let’s assume first that because the inorganic compound in bones, the Hydroxylapatite (Calcium & Phosphorous element combination) which causes the bone to have it’s intrinsic quality of being so tough and hard, with a compressive and tensile strength in the level of stainless steel. Let’s say for argument that if the hypertrophy of chondrocytes in the epiphysis was not enough to cause long bone lengthening through hydrostatic pressure and expansion of the trabecular and cortical bone, we would then need to find another way to cause the hard bone to separate, at least long enough for chondrocytes and cartilage to get between the hard inorganic spaces to push them apart. This is way I have almost always believed that besides the LSJL method which seems promising, there may be no way to cause adult height increase in a non-invasive way. I thought that a bone distraction or fracture was needed. However, after going through the literature recently I have concluded instead that maybe it is possible if we can use some form of process to demineralize the Hydroxylapatite from the bone matrix at a level where the tensile strength of the bone is dramatically decreased. I remember reading from HeightQuest that Tyler suspected that the Ginza Kojima machine operated on this principle where the green light emitted or the chemical bath used was to somehow demineralize the bone and give it a lower young’s modulus value. Then a torsional or twisting load was made to the bone since the torsional loading value of long bones with the cortical bone was not that high. From what Tyler, Sky, and I have seen the machine has been around on the market for many years now with little news or press placed on it. In my opinion, mineralization may be a key step that might be required to be added on for the entire height increase technique to work. There might indeed be a step where limbs are subjected to some form of chemical, electrical stimuli, or other to weaken the bones a little.

Analysis: In the study the researchers used a suitable method for rapid demineralization of mouse teeth in 0.1 M (molar) EDTA (ethylene diamine tetra-acetic acid) at 42˚C . From the study the mice were eventually killed to study the bone mineralization so obviously this type of testing can’t be done on human subjects but the major take away should be that we can try these methods where the subject’s limb is placed in a weak acid mixture, have the heat increased, (maybe not as high as 42 C) and slowly over time have the bones demineralized for possible tensile loading later. It has already been reported that bone demineralization was accelerated by microwave treatment to heat the subjected bone up and increased the rate of dimeralization by 2X, or at least with tested mice skulls. In the study, the researchers showed that there are plenty of stronger acids out there that can do the demineralization work a lot better and faster but that woudl destroy the genes and gene expression of the underlying body tissue, which would include for a live human subject muscle and bone and ligaments. We obviously want to avoid that. It would require more research to find a better acid solution that can extract bone minerals while keeping most other tissues safe and intact.

Overall, this is my first crude attempt at trying to find a way to dimineralize the bones. I would suspect for future testing, it may be a better idea to focus on just a small 4-5 mm thick layer of axial directional bone and try to dimineralize that. area. If that that process is successful, we can either induce chondrocytes to replace those areas where the hydroxylapatite was or increase the activity of osteoclasts to remove the bone material for growth factors and progenitor chondrogenetic cells.

From source link HERE…

A Method for Rapid Demineralization of Teeth and Bones

Andrew Cho, Shigeki Suzuki, Junko Hatakeyama, Naoto Haruyama, and Ashok B. Kulkarni

Abstract: Tooth and bone specimen require extensive demineralization for careful analysis of cell morphology, as well as gene and protein expression levels. The LacZ gene, which encodes the ß-galactosidase enzyme, is often used as a reporter gene to study gene-structure function, tissue-specific expression by a promoter, cell lineage and fate. This reporter gene is particularly useful for analyzing the spatial and temporal gene expression pattern, by expressing the LacZ gene under the control of a promoter of interest. To analyze LacZ activity, and the expression of other genes and their protein products in teeth and bones, it is necessary to carry out a complete demineralization of the specimen before cutting sections. However, strong acids, such as formic acid used for tooth demineralization, destroy the activities of enzymes including those of ß-galactosidase. Therefore, most protocols currently use mild acids such as 0.1 M ethylene diamine tetra-acetic acid (EDTA) for demineralization of tooth and bone specimen, which require a longer period of treatment for complete demineralization. A method by which hard tissue specimens such as teeth and bones can be rapidly, but gently, decalcified is necessary to save time and effort. Here, we report a suitable method for rapid demineralization of mouse teeth in 0.1 M EDTA at 42˚C without any loss of ß-galactosidase activity.

INTRODUCTION

Teeth are the hardest tissues in the body, consisting of enamel, dentin, and cementum, and have a highly mineralized extracellular matrix. Unlike soft tissues, which can be easily sectioned and analyzed, teeth require complete demineralization in order to prepare proper sections for histological
analysis using certain stains and antibodies. In the past, many decalcifiers have been tested for different purposes. For example, the Morse’s solution, a strong acid baseddecalcifier, has been used for the rapid detection of RNA by in-situ hybridization (ISH) [1]. Another acid-based solution,
5% trichloroacetic acid (TCA), has been used for the analysis of DNA strand breaks by terminal deoxy (d)-UTP nickend labeling (Tunnel) [2]. EDTA containing solution is a mild demineralization agent which has been mostly used for immunolocalization studies because of its antigen-preserving
properties. Treatment of bones with 0.1 M EDTA following initial fixation in glutaraldehyde has been reported as adequate for immunocytochemical localization of certain bone matrix proteins [3]. Furthermore, the treatment of dental tissues with 4.3% EDTA resulted in satisfactory preservation of the fine structures of the cells and matrices [4], however it requires a relatively long incubation time in order to achieve complete demineralization [5,6]. For example, to demineralize human deciduous teeth for analysis, it requires 4 weeks of demineralization using 10% EDTA [7]. To demineralize a 1-month-old mouse skull with similar EDTA solution it takes about 3-4 weeks of treatment to prepare the specimen for analyzing expression levels of the LacZ and growth fac

*Address correspondence to this author at the Chief, FGS, CDBRB, NIDCR, NIH, Tel: 301-435-2887; Fax: 301-435-288; E-mail: ak40m@nih.gov tor genes [8,9].

To achieve fast demineralization, a microwave-induced demineralization method has been examined [10,11]. While this method has achieved faster demineralization time compared to the traditional method, preserved the fine morphological structures of the bone tissue, and retained RNA, its effect on LacZ gene expression has not been addressed. Moreover it has been speculated that the advantages associated with using the method are due to the higher temperature attained during microwaving the specimen purely due to thermal effect [12]. The LacZ gene is widely used to investigate promoter activity [13-15], however there is not detailed protocol for optimal analysis of ß-galactosidase activity in decalcified mouse teeth and bones. Moreover, the effects of acid-based decalcifiers and different fixatives used for preparation of mouse teeth specimen to analyze ß-galactosidase activity have not been well established. In this report, we demonstrate the effects of several fixatives and decalcifiers on mouse teeth specimen for LacZ activity, whose expression is driven by a tooth specific DSPP (Dentin Sialophophoprotein) promoter. Most importantly, we have explored the effects of elevated temperatures on mouse teeth specimen to optimize a rapid but gentle demineralization method suitable for analysis of LacZ activity. This will prove to be a suitable method for rapid demineralization of mouse skulls for analysis using histochemical staining and in-situ hybridization.

DISCUSSION 

We set out to optimize the demineralization method in order to make it shorter and safer for histological analysis of mouse teeth. In the process of this optimization, we addressed some of the key issues related to the effects of the decalcifiers and fixatives used for preparing the mouse skull specimen. Our study indicates that the demineralization of mouse skulls at 42°C using 0.1M EDTA can significantly shorten the time required for complete demineralization, while also retaining sufficient ß-galactosidase activity. DSPP, a key dentin extracellular matrix protein secreted by odontoblasts, plays a crucial role in the mineralization of predentin to form mature dentin [18-21]. We previously reported the isolation and characterization of the murine Dspp gene [22] and the validation of its promoter sequence [16]. The Dspp-LacZ transgenic mice used in the present study to display spatial and temporal expression patterns similar to endogenous DSPP expression profiles [16]. In these mice, the LacZ gene, which is placed under the DSPP promoter sequence, is expressed as early as embryonic day 17.5 (E17.5) in preameloblasts and odontoblasts, and at E18.5 in molars. Its expression in preameloblasts is transient, whereas it stays robust in odontoblasts throughout tooth development. Because of this expression profile, the tooth specimens from these mice are ideal for testing the effects of decalcifiers, fixatives and incubation conditions on the stability of ß-galactosidase activity. However, because the LacZ gene, which encodes ß-galactosidase, has been widely used as a reporter gene to analyze the expression of a gene of interest, the demineralization protocol that we have optimized in the current study can be applied to other transgenic mouse models expressing ß-galactosidase in teeth and bones. We first verified the effects of various fixatives and the fixation time on the stability of the ß-galactosidase enzyme in the tooth sections. We tested 0.25% glutaraldehyde, 4% PFA, zinc formalin and formalin for their effects, and found that all of them work well as fixatives to analyze ß-galactosidase activity in the tooth sections (Fig. 1). As for the fixation time, we found that post-incubation of the skulls with 4% PFA for up to an hour did not have any adverse effects on ß-galactosidase activity. However, extending the incubation time to 2 hours adversely affected the enzyme activity (Fig. 2). As previously reported [17], LacZ enzymatic activity in kidney tissue was also retained during a 1- hour incubation time with 4% PFA, but not during a longer period of time.

Next, we tested the acid-based demineralization reagents, Formical-4 and Immunocal, in order to analyze their effects on ß-galactosidase activity. As you can see in Fig. (3), it is clear that 0.1% EDTA, but not the acid-based demineralization reagents, maintained ß-galactosidase enzyme activity after complete demineralization (Fig. 3). Chelating agents such as EDTA can bind to calcium and other ions and remove them from mineralized tissue. This reaction occurs gently, and enzymatic activity can therefore be retained during demineralization. In contrast, long-term exposure to acidic conditions could potentially destroy enzymatic activity such as ß-galactosidase. Another possibility is that, because Formical-4 and Immunocal contain fixatives, ß-galactosidase may be over-fixed and thus result in a lack of enzymatic activity. As shown in Fig. (4), we then explored the potential for shortening the demineralization process by increasing the temperature during demineralization, since it has already been reported that bone demineralization was accelerated by microwave treatment [10,11]. Our data indicated that incubation of mouse skulls with 0.1M EDTA at 42°C results in demineralization that is 2-fold faster than incubation at room temperature. Our data also revealed that, although an incubation temperature of 50˚C can achieve demineralization the shortest incubation time, it adversely affects the enzyme activity (Fig. 5). These data suggest that incubation at 42°C is most useful for analysis of enzymatic activity in decalcified teeth. Moreover, as shown in Fig. (6), 6 days of incubation with 0.1M EDTA at 42°C was sufficient for complete demineralization of 1-year-old mouse teeth. These observations suggest that any specimens can be demineralized by incubation with 0.1M EDTA at 42°C for 6 days.

To assess the effects of elevated temperatures on the stability of nucleic acids, we extracted total RNA from the tooth sections of the skulls subjected to different temperatures for the demineralization process, and performed RTPCR analysis. The amplification of Gapdh mRNA transcripts at 37°C, 42°C or 50°C showed similar levels, indicating that a higher temperature had no affect on the stability of nucleic acids (Fig. 7).

In summary, we have optimized the conditions for rapid demineralization of mouse skulls, which significantly shortens the time required for complete demineralization but does not affect the enzyme activity or nucleic acids in the tooth sections. We recommend 0.1M EDTA as a safe decalcifier, to be used at 42°C for successful demineralization of mouse skulls. These conditions could be widely applied for the enzymatic and immunohistochemical analysis of other proteins in hard tissues. Moreover, rapid demineralization using this method will promote quicker analysis of genes and proteins implicated in tooth and bone diseases.

ACKNOWLEDGEMENTS

We would like to thank Dr. Taduru Sreenath for his helpful suggestions during the course of this project. We also would like to thank Drs. Marian Young and Larry Fisher for critical reading of the manuscript, and Shelagh Powers for expert editorial corrections. This work was supported by funds from the Division of Intramural Research of the National Institute of Dental and Craniofacial Research.

{Update: It seems that from the main website AyurvedicUrea.com, the main ingredient found in ayurvedic urea is “Dhatrumurgasiniy”. It seems this thing can only be found in Nepal. I will be doing some more research on what this plant/ compound really is.]

What I wanted to do with this post is try to prove conclusively that this new idea for height increase is either something worth looking into or just the newest craze in height increase to com along. When I typed in the words “Ayurvedic Urea Height Increase” into Google, I was surprised at what else I managed to find.

There is a Topix.com Discussion HERE…

From the first looks of it, it would appear that at least a dozen people have all said that it is a real product which worked for them. This seems great until you realize that all the people who said that the product is legitimate have their location traced back to one location, Lalitpur, Nepal. Even what we would say is american names like Bob are coming from the same place. There seems to be another person from the UK also saying it is legitimate so we have two people who are saying it works. Most of the other posters are not sure or claim it is a scam.

From AyurvedicUrea.com…

This website is the official website for the idea to increase height using ayurvedic urea. It seems that this website is based off of San Francisco from the Facebook page profile. They say on their page for scams that you should not buy “fake” ayurvedic urea from any providers from Africa or any one else EXCEPT them. So they seem to want you to only use them as a service. Hm….That makes me suspicious. When an organization is trying this hard to push out any possible competitors by claiming everyone else is a liar and scam artist, that make you wonder if they are a scam too. Something else to note is that the website seems to be based off of the Blogger Blogging Platform. That makes it seem like it is not very professional. However, I would have to concede that this website was built on a blogging platform too, WordPress. From this link HERE these guys say that they are the only people who sell the real Ayurvedic Urea. Are these sellers saying that they are the only people who can make this stuff? The website lists two other websites being fake sites selling fake ayurvedic urea however when I copy and pasted the URL domain names, I got only a standard WordPress 2011 theme with maybe 2 pages of weak content. Sites are www.buyayurvedicurea.com and www.ayurvedicurea.net. If you want to see what is on the websites, go ahead. There is almost nothing on these sites that the other site says makes them a scam. It’s Nothing. From the website there seems to be one “legitimate” sellers of the stuff. NAME: MELINA JATAMARI    CONTACT EMAIL: melinajatamari@gmail.com. Apparently she is the “only seller we have verified and authorized“. What the website does seem to do is hold the money in escrow so that the money you give will only reach them after you have tested the compound and seen its effects. The urea doesn’t seem to work on dwarves. There is research being conducted in Singapore starting around 11//23/2012 and will end in 7/23/2013. The next group of people being tested will be in Thailand. If you want to contact these people email them at support@ayurvedicurea.com. Or get in touch with some women named Sharon Stone (the same as the actress??) Here is the pricing from the website…

From a WordPress blog based website MeetMeDaily.com…

This website seems to be some form of funnel or sales page to get the person to go to the Ayurvedic Urea website. There is absolutely no scientific information on why this product would even work. There is even a Facebook page for Ayurvedic Urea located HERE. From the first look, there is over 6000 likes, and the profile website was created in 2009. The biggest discussion has around 54 comments with most of the profiles saying the product worked on them. However after I clicked on the profiles It showed that the profiles had no pictures, information, or friends. This seems to suggest that someone purposely created a few dozen fake Facebook Profiles to write up fake positive testimonials for this Ayurvedic Urea. There were real profiles who commented to the Facebook page asking how they can buy this extremely expensive product but it seems that no one has answered them back yet.

From AGRHood.com…