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brundel
09-22-2011, 06:28 PM
Sci Transl Med 11 November 2009:
Vol. 1, Issue 6, p. 6ra15
DOI: 10.1126/scitranslmed.3000112
• Research Article
Gene Therapy
Follistatin Gene Delivery Enhances Muscle Growth and Strength in Nonhuman Primates


Abstract
Antagonists of myostatin, a blood-borne negative regulator of muscle growth produced in muscle cells, have shown considerable promise for enhancing muscle mass and strength in rodent studies and could serve as potential therapeutic agents for human muscle diseases. One of the most potent of these agents, follistatin, is both safe and effective in mice, but similar tests have not been performed in nonhuman primates. To assess this important criterion for clinical translation, we tested an alternatively spliced form of human follistatin that affects skeletal muscle but that has only minimal effects on nonmuscle cells. When injected into the quadriceps of cynomolgus macaque monkeys, a follistatin isoform expressed from an adeno-associated virus serotype 1 vector, AAV1-FS344, induced pronounced and durable increases in muscle size and strength. Long-term expression of the transgene did not produce any abnormal changes in the morphology or function of key organs, indicating the safety of gene delivery by intramuscular injection of an AAV1 vector. Our results, together with the findings in mice, suggest that therapy with AAV1-FS344 may improve muscle mass and function in patients with certain degenerative muscle disorders.

o Received April 22, 2009.
o Accepted October 19, 2009.
• Copyright © 2009, American Association for the Advancement of Science
Citation: J. Kota, C. R. Handy, A. M. Haidet, C. L. Montgomery, A. Eagle, L. R. Rodino-Klapac, D. Tucker, C. J. Shilling, W. R. Therlfall, C. M. Walker, S. E. Weisbrode, P. M. Janssen, K. R. Clark, Z. Sahenk, J. R. Mendell, B. K. Kaspar, Follistatin Gene Delivery Enhances Muscle Growth and Strength in Nonhuman Primates. Sci. Transl. Med. 1, 6ra15 (2009).

brundel
09-22-2011, 06:32 PM
LONG-TERM MUSCLE IMPROVEMENTS SHOWN IN GENE THERAPY STUDY IN MICE

COLUMBUS, Ohio – Injecting a gene responsible for making a specific protein into a mouse that’s used as a model for muscular dystrophy can lead to long-term improvements in the animal’s muscle size and strength, a new study shows.
Researchers investigating the gene delivery of the protein in animals suggest the results warrant testing the same approach in human clinical trials for diseases associated with muscle wasting, including Duchenne muscular dystrophy, the most common form of the childhood disorder.


Scientists used a safe virus to deliver a protein called follistatin into the leg muscles of young and older mice that have a disorder similar to human Duchenne muscular dystrophy (DMD). The protein inhibits the activity of myostatin, identified in previous research as a protein that limits muscle growth. Both young and old mice treated with the therapy responded with increased muscle mass and improvements in strength.

The study also takes a rare long look at the effects of the therapy.
“Many studies don’t evaluate a therapy over a two-year time span. In our studies, the beneficial effects persisted over the two years we evaluated,” Kaspar said. “Furthermore, this long-term study shows that there were no obvious safety problems with either the gene therapy virus or the therapeutic protein, follistatin.”
The research is reported online in this week’s edition of Proceedings of the National Academy of Sciences.

In studies of younger mdx mice, the therapy was administered when they were 3 weeks old. At age 5 months, they had a larger body mass and higher muscle weight than did comparison animals.
Mice used for comparison were treated with an inactive fluorescent protein that allowed researchers to monitor which cells were affected by the experimental gene therapy technique.
Before testing follistatin in mdx mice, the scientists first tested the protein in normal mice and found after 725 days that they, too, had increased muscle mass and better grip strength when compared to untreated mice.
The resulting muscle enhancements in all of the treated mice were evident at the site of injection as well as in tricep muscles, meaning the therapy was able to affect other muscle cells in the body. To deliver the protein, scientists used an adeno-associated virus that had been manipulated to find its way into target cells without promoting any spread of the virus itself.


This research was supported by Project A.L.S., the Myositis Association, Roger Stevens and a National Research Service Award fellowship.

brundel
09-22-2011, 06:34 PM
Am J Physiol Endocrinol Metab. 2011 Mar;300(3):E543-53. Epub 2011 Jan 4.

Follistatin-derived peptide expression in muscle decreases adipose tissue mass and prevents hepatic steatosis.

Nakatani M, Kokubo M, Ohsawa Y, Sunada Y, Tsuchida K.
Source
Division for Therapies against Intractable Diseases, Institute for Comprehensive Medical Science (ICMS), Fujita Health University, Toyoake, Aichi, Japan.
Abstract
Myostatin, a member of the transforming growth factor (TGF)-β superfamily, plays a potent inhibitory role in regulating skeletal muscle mass. Inhibition of myostatin by gene disruption, transgenic (Tg) expression of myostatin propeptide, or injection of propeptide or myostatin antibodies causes a widespread increase in skeletal muscle mass. Several peptides, in addition to myostatin propeptide and myostatin antibodies, can bind directly to and neutralize the activity of myostatin. These include follistatin and follistatin-related gene. Overexpression of follistatin or follistatin-related gene in mice increased the muscle mass as in myostatin knockout mice. Follistatin binds to myostatin but also binds to and inhibits other members of the TGF-β superfamily, notably activins. Therefore, follistatin regulates both myostatin and activins in vivo. We previously reported the development and characterization of several follistatin-derived peptides, including FS I-I (Nakatani M, Takehara Y, Sugino H, Matsumoto M, Hashimoto O, Hasegawa Y, Murakami T, Uezumi A, Takeda S, Noji S, Sunada Y, Tsuchida K. FASEB J 22: 477-487, 2008). FS I-I retained myostatin-inhibitory activity without affecting the bioactivity of activins. Here, we found that inhibition of myostatin increases skeletal muscle mass and decreases fat accumulation in FS I-I Tg mice. FS I-I Tg mice also showed decreased fat accumulation even on a control diet. Interestingly, the adipocytes in FS I-I Tg mice were much smaller than those of wild-type mice. Furthermore, FS I-I Tg mice were resistant to high-fat diet-induced obesity and hepatic steatosis and had lower hepatic fatty acid levels and altered fatty acid composition compared with control mice. FS I-I Tg mice have improved glucose tolerance when placed on a high-fat diet. These data indicate that inhibiting myostatin with a follistatin-derived peptide provides a novel therapeutic option to decrease adipocyte size, prevent obesity and hepatic steatosis, and improve glucose tolerance.
PMID:
21205933
[PubMed - indexed for MEDLINE]

brundel
09-22-2011, 06:36 PM
Regulation of myostatin activity and muscle growth

Se-Jin Lee* and Alexandra C. McPherron
Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205
*To whom reprint requests should be addressed. E-mail: sjlee@jhmi.edu.
Communicated by David L. Garbers, University of Texas Southwestern Medical Center, Dallas, TX
Received March 26, 2001; Accepted May 30, 2001.

ABSTRACT
Myostatin is a transforming growth factor-β family member that acts as a negative regulator of skeletal muscle mass. To identify possible myostatin inhibitors that may have applications for promoting muscle growth, we investigated the regulation of myostatin signaling. Myostatin protein purified from mammalian cells consisted of a noncovalently held complex of the N-terminal propeptide and a disulfide-linked dimer of C-terminal fragments. The purified C-terminal myostatin dimer was capable of binding the activin type II receptors, Act RIIB and, to a lesser extent, Act RIIA. Binding of myostatin to Act RIIB could be inhibited by the activin-binding protein follistatin and, at higher concentrations, by the myostatin propeptide. To determine the functional significance of these interactions in vivo, we generated transgenic mice expressing high levels of the propeptide, follistatin, or a dominant-negative form of Act RIIB by using a skeletal muscle-specific promoter. Independent transgenic mouse lines for each construct exhibited dramatic increases in muscle mass comparable to those seen in myostatin knockout mice. Our findings suggest that the propeptide, follistatin, or other molecules that block signaling through this pathway may be useful agents for enhancing muscle growth for both human therapeutic and agricultural applications

The most dramatic effects on skeletal muscle were obtained by using the follistatin construct. We obtained two founder animals (F3 and F66) that showed increased muscling. In one of these animals (F3), muscle weights were increased by 194–327% relative to control animals, resulting from a combination of hyperplasia (66% increase in fiber number to 13,051 in the gastrocnemius/plantaris) and hypertrophy (28% increase in fiber diameter to 55 μm). Although we have not analyzed muscle weights of myostatin knockout mice in a hybrid SJL/C57BL/6 background, the increases in muscle mass observed in the F3 founder animal were significantly greater than the increases we have seen in myostatin null animals in other genetic backgrounds (unpublished results; see also ref. 1). These results suggest that at least part of the effect of follistatin may result from inhibition of another ligand besides myostatin. Clearly, analysis of additional follistatin transgenic lines will be essential in determining whether other ligands may also be involved in negatively regulating muscle growth.

brundel
09-22-2011, 06:37 PM
http://i2.photobucket.com/albums/y26/brundel/follistatin1.jpg

brundel
09-22-2011, 06:54 PM
In human studies run mainly by bodybuilders like you and me Follistatin has been shown to be exceptionally effective at causing muscle hypertrophy.
Most users site gains in the range of 1lb muscle per day.
These gains seem at this time to be persistent meaning that unlike most AAS that we would expect this type of mass gain from the weight gain is from actual muscle tissue and not water and glycogen. Therefore, after cessation of treatment with follistatin one can expect to retain weight gain attained while on cycle.

Follistatin can be very irritating at the injection site for some users.
Keep in mind Follistatin is NOT FDA approved for human use and no clinical human trails have been run to date.

TGB1987
09-22-2011, 07:41 PM
Very Very interesting Brundel this sounds like a wonder drug. The pics of the mouse were shocking. I would love to see what this can do in Humans. Thanks for the great information.

brundel
09-22-2011, 07:44 PM
There are alot of guys running follistatin and the results are pretty dramatic.
1lb muscle gain per day seems to be about the norm which puts it easily on par with the strongest AAS.

Digitalash
09-22-2011, 08:29 PM
It does sound pretty amazing, I'd imagine the viral version will find it's way onto the market at some point and god only knows what will happen then. Last time I was researching it everyone was saying it destroys tendons and if you used it you'd be fucked for life, but this research seems to say otherwise.

I can't wait to see the kind of physiques that will come along when the pro's get a hold of this. It would probably make GH/slin obsolete so hopefully there'll be a return to the smaller waists of Arnold's era but with as much or more mass than the pros of today.

brundel
09-22-2011, 08:33 PM
I have not found any research to support the theory that it destroys tendons.
Granted there are likely some sides we have yet to discover but for now it seems pretty safe in short bursts.

Digitalash
09-22-2011, 10:09 PM
I have not found any research to support the theory that it destroys tendons.
Granted there are likely some sides we have yet to discover but for now it seems pretty safe in short bursts.

Yeah I don't see much happening in 10 days even if that is the case, I'm thinking more about the viral version. Not gonna lie if I had access to it I'd give it serious thought...

DLEATO
09-23-2011, 03:10 AM
Its not that it destroys tendons, but more so that strength increases but tendons still stay the same.


if you used it you'd be fucked for life

where did this come from?
(not disagreeing with you in any way, just wanna read the whys)

dirtwarrior
09-23-2011, 04:12 AM
Its not that it destroys tendons, but more so that strength increases but tendons still stay the same.

truth

Where can get legit stuff for a decent price

brundel
09-23-2011, 08:51 AM
truth

Where can get legit stuff for a decent price

Click on the link in my sig.

brundel
09-23-2011, 08:55 AM
It doesnt seem that there is an excessive increase in strength like with an AAS known for strength. It does seem that strength is consistent with new muscle tissue growth and it is not likely to cause tendon damage with 1 cycle.

Digitalash
09-23-2011, 12:40 PM
Its not that it destroys tendons, but more so that strength increases but tendons still stay the same.



where did this come from?
(not disagreeing with you in any way, just wanna read the whys)


I don't remember where I read it but it was more referring to the viral version, ie. you inject it once and your genes are changed forever. I haven't looked into it in a long time and there wasn't much info back then but there was something about it negatively affecting crosslinking in tendons making them weaker and more brittle. These studies seem to be new and there's no mention of it so I'm guessing that was just a theory or based on old research.

So even if it does have an impact on tendons, which I'm not saying it does, but with a short cycle it shouldn't be any problem. If you got a hold of the viral version though and that were the case you could end up with serious issues somewhere down the line. Sooo I'd be pretty hesitant to use it until there's alot more research done, because once you did there's no turning back

DLEATO
09-23-2011, 01:04 PM
ohk, that makes more sense.I was wondering how that would be possible with a compound that would be out your system so quick. Apparently your only suppose to run it for 10 days, but i have heard of people running it for a month.

My folli's on its way so you should hear some personal feedback soon.

dirtwarrior
09-23-2011, 01:45 PM
Is it systemic or just for isolated muscle growth?

TwisT
09-23-2011, 01:46 PM
myostatin inhibitors have been proven to destroy ligaments and tendons in research, saying they don't is simply incorrect.

Heres one study, there are many more I dont feel like posting. I'm a huge fan of folli, but I dont want anyone mislead in any way.
___________________________

Tendons of myostatin-deficient mice are small, brittle, and hypocellular.
Mendias CL, Bakhurin KI, Faulkner JA. Source Department of Molecular and Integrative Physiology, University of Michigan Medical School, Ann Arbor, MI 48109, USA.

Abstract

Tendons play a significant role in the modulation of forces transmitted between bones and skeletal muscles and consequently protect muscle fibers from contraction-induced, or high-strain, injuries. Myostatin (GDF-8) is a negative regulator of muscle mass. Inhibition of myostatin not only increases the mass and maximum isometric force of muscles, but also increases the susceptibility of muscle fibers to contraction-induced injury. We hypothesized that myostatin would regulate the morphology and mechanical properties of tendons. The expression of myostatin and the myostatin receptors ACVR2B and ACVRB was detectable in tendons. Surprisingly, compared with wild type (MSTN(+/+)) mice, the tendons of myostatin-null mice (MSTN(-/-)) were smaller and had a decrease in fibroblast density and a decrease in the expression of type I collagen. Tendons of MSTN(-/-) mice also had a decrease in the expression of two genes that promote tendon fibroblast proliferation: scleraxis and tenomodulin. Treatment of tendon fibroblasts with myostatin activated the p38 MAPK and Smad2/3 signaling cascades, increased cell proliferation, and increased the expression of type I collagen, scleraxis, and tenomodulin. Compared with the tendons of MSTN(+/+) mice, the mechanical properties of tibialis anterior tendons from MSTN(-/-) mice had a greater peak stress, a lower peak strain, and increased stiffness. We conclude that, in addition to the regulation of muscle mass and force, myostatin regulates the structure and function of tendon tissues.

cottonmouth
09-23-2011, 01:50 PM
Damn that rat is thick!

The viral version sounds to great to be true though.

But it might be like those animals with double muscle mutation. The Belgium blue bull or something like that and then there is a dog that it happens in,, Its called a bully somehting or other. cant think of the name.

But they suffer from no ill effects.

brundel
09-24-2011, 05:31 PM
The blue bulls you are thinking about are knockout specimens.
They were either born without or genetically altered to be born without myostatin.
The rats in the photos I posted had myostatin but it was inhibited with follistatin.

These examples were born with these genetic alterations. The same result is not expected obviously with short burts cycles of follistatin. There were test subjects with 300%+ increase in muscle tissue, for our purposes and uses and in real life human tests thus far it seems as though about 1lb a day is pretty normal. Im not sure what would happen if you ran it for 300 days.....I dont think I would want to be the guy to find out.

dirtwarrior
09-24-2011, 06:22 PM
For muscle growth does it have to be injected into each muscle group?

brundel
09-24-2011, 07:50 PM
Alot of people seem to be shooting it into or near the muscle and getting localized muscle growth. In test studies muscle growth was seen at the site of injection but also apparent at other locations.

"The resulting muscle enhancements in all of the treated mice were evident at the site of injection as well as in tricep muscles, meaning the therapy was able to affect other muscle cells in the body"

KUVinny
09-24-2011, 08:20 PM
BW is sufficient for reconstitution correct?

And what about storage? Refrigerate?

Thanks brundel for all the info!

keith1569
09-24-2011, 08:44 PM
truth

Where can get legit stuff for a decent price


You can also get some from ergo pep..they will price match if u find a lower price

keith1569
09-24-2011, 08:45 PM
BW is sufficient for reconstitution correct?

And what about storage? Refrigerate?

Thanks brundel for all the info!


Ya you recon with BAC water and store in the fridge..it starts to degrade around 7 days

KUVinny
09-24-2011, 08:58 PM
Thanks.

Erinn
11-15-2011, 02:12 AM
I have seen the before and after results and i have to say...2 thumbs up

cottonmouth
11-16-2011, 12:23 PM
imagine using the follistatin with slin, gh, and aas.

sounds like the holy grail of muscle growth.

brundel
11-16-2011, 01:31 PM
I have seen the before and after results and i have to say...2 thumbs up

What you saw was actually 200mcg GHRP-6 3x daily.
The benefits were obvious because one of my major issues is getting in adequate cals to gain weight. Real GHRP-6 makes me painfully hungry.
It sucks because most online suppliers sell bunk peptides. Im sad my source is gone after all this bullshit.