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My GHRP-2 CJC-1295 thread

Anabolic Steroids Discuss My GHRP-2 CJC-1295 thread in the Steroid forums; Just gonna start a new thread with this Hi Im gonna give this a try for three months when the ...

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Old 06-07-2010, 02:57 AM
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Default My GHRP-2 CJC-1295 thread

Just gonna start a new thread with this

Hi
Im gonna give this a try for three months when the goods gets here could be another week yet.
GHRP-2 CJC-1295 without Dac
A once a day dose before bed to try and get the youthful amounts of GH back.
Things I will be looking out for are skin elasticity and BF%.
Dose will be 200mcg GHRP-2 and 100mcg of CJC-1295 pre-bed every night

Here is some info I gathered together while researching this.


Dosing
1
In the studies on growth hormone releasing hormone (GHRH) and all the different growth hormone releasing peptides (GHRP-6, GHRP-2, Hexarelin...) they either use a weight based dosing, for example 1mcg/kg or just fixed amount, for example 100mcg.

For some reason I have never seen a study indicate that the distinction in methodology mattered. My best guess is that a large portion of the growth hormone studies are aimed at children. Children w/ growth hormone defeciency were the first approved category of prescribed users and they continue to be the largest target group. Weight and drug use matters in children. Drug reactions both good & bad are positively correlated to weight in children. There is much more concern about the body's ability to compensate for too much of an administered drug. Adults seem to have more tolerance to a wider range of drug dosing.

My present opinion is that weight should not be used to determine adult dosages. Gender differences of course...but this is not related to weight. So if you are 200 pounds or 250 pounds it shouldn't matter. Again this is just my opinion because I haven't ever found anything to indicate that it is important.

Of course absence of knowledge is not knowlede of absence so take it for what its worth.

I believe that a bodybuilding dose is:

100mcg of CJC-1295 + 100 - 200 mcg GHRP-6 three times a day.

...and an anti-aging dose (still high enough for fatloss) is 100mcg of CJC-1295 pre-bed and 100mcg of GHRP-6 twice a day.

2
Quote:
Originally Posted by kutch
Also, any opinions on this study:
50mcg of CJC + 50mcg of GHRP-6 daily AND additional 50mcg of GHRP-6 PWO(or pre-breakfast)
You will get an increase in GH levels at that dose. If you do that run it for a longer period of time...and you should see a contribution to a tighter core.
Quote:
Originally Posted by kutch
Learning a lot here. Are you saying to dose a CJC/GHRP combo pre-bed?
Yes.
Quote:
Originally Posted by kutch
Others have said that dosing hgh pre-bed may affect your normal hgh release after you fall asleep. Is the CJC/GHRP combo different since your body is producing the hgh and it's not coming from an external source which may trigger your body to stop producing hgh?
Yes.
Quote:
Originally Posted by kutch
Also, since your body is already producing an hgh spike after you sleep, wouldn't it be more cost effective(if that's a priority) to skip that pre-bed dose?
No. In males the night-time growth hormone pulse is many many multiples larger then at any other time of the day.

So using an arbitrary example, if you introduce a doubling agent (i.e. a compound/factor capable of doubling the effect/outcome) into a pulse that has a secretory value of 100 then you end up with 200.

However if you introduce that same doubling agent into a pulse that has a secretory value of 10 then you end up with 20.

Aside from just mere volume the night-time growth hormone release is positively correlated with slow wave sleep. If you can do something to support slow wave sleep then you also end up supporting strong GH release. If you can do something to support night-time GH release you also end up supporting restful "growth & repair" promoting sleep.

Thats why when you use CJC or GHRP-6 you end up with deeper more regenerative sleep.

Below is something I wrote explaining Slow Wave Sleep & GH release. I know it is not directly related to your question but it is helpful to understand:
SWS & GH release

There are two types of sleep, rapid eye movement (REM) and non-rapid eye movement (NREM). Sleep proceeds in cycles composed of four types of stages of NREM and a stage of REM usually ordered as: 1 > 2 > 3 > 4 > 3 > 2 > REM

The cycle lasts on average 90 to 110 minutes, with a greater quantity of stages 3 and 4 experienced early in the night and more REM later in the night.

NREM accounts for 75–80% of total sleep time. Non-REM is comprised of four stages; stages 1 and 2 are considered 'light sleep', and 3 and 4 'deep sleep' or slow-wave sleep (SWS).

It has been shown that sleep, more specifically slow-wave sleep (SWS), does affect growth hormone levels in adult men. During eight hours sleep, it has been demonstrated in several studies that the men with a high percentage of SWS (average 24%) also had high growth hormone secretion, while subjects with a low percentage of SWS (average 9%) had low growth hormone secretion.

In one very complete study referenced by several others, it was demonstrated that “GH secretory rates and peripheral GH concentrations were maximally correlated with sleep stage, with lags of 4.5 and 16 min, respectively, suggesting that maximal GH release occurs within minutes of the onset of stage 3 or 4 sleep”.

Furthermore “sleep-related augmentation of GH secretion… usually occurs around midnight and the GH levels at that time are, as a rule, at their highest during the 24-hour period. Partially, this phenomenon is time-entrained and partially related to sleep itself. It is associated with a slow wave sleep, and the maximal GH levels occur within minutes of the onset of slow wave sleep” -Holl RW, Hartman ML, Veldhuis JD, et al. Thirty-second sampling of plasma growth hormone in man: correlation with sleep stages. J Clin Endocrinol Metab 1991;72:854–61.

The origin of nocturnal GH release in humans is still unknown. Most likely hypothalamic GHRH release is a major contributing component, but an additional role of another factor, presumably augmenting GHRH responsiveness of the somatotrophs, is likely. However the precise explanatory mechanisms are still not fully identified.

It is worth reiterating though that nocturnal release of GH makes up only a fraction of the total daily GH release in women, but the bulk of GH output in men.
__________________

3
On the topic of GHRP-6 dosing [Here is the range]

Assuming that your GHRP-6 (or any of the GHRPs (i.e. GHRP-2, Hexarelin...) is of the same quality as that used in the studies then 100mcg is enough.

The saturation dose was determined to be 100mcg. So the studies that use GHRP-6 for the most part used either 100mcg or 1mcg/1kg of bodyweight. Consequently most of the GH release numbers for GHRP-6 that we discussed in this thread came from studies on humans dosing 100mcg at a time.

However it has been determined in a few studies, particularly the ones using Hexarelin as the GHRP that the highest dosing after which there is no effect is somewhat variable among people and could be 200mcg to 400mcg.

On the otherhand there has been demonstrated synergy in GH (growth hormone) release between GHRH (growth hormone releasing hormone) and GHRP-2 (growth hormone releasing peptides) at the following dose: 100mcg GHRH + 30mcg GHRP-2

4
After I finish this cycle I'm dropping down to just 100mcg of CJC-1295 pre-bed w/ GHRP-6. That's only 700mcg per week and that is more than enough to derive a solid longer-term benefit.

I've been on the higher dosing scheme for several weeks with several more weeks to go and I "feel" for me that it may be too high off cycle.

So the lower dose you mentioned need not be considered a "poor man's" dose especially if you use a spot of GHRP-6 (or Hex or GHRP-2 or Ipamorelin) with it.

By-the-way you also proposed a 5 day on/ 2 off protocol with 100mcg of CJC-1295 used at each dosing. The overall levels would look like the numbers I used in the post above except they would be half (50%) of those values.

5
Sub Q or intramuscular
Yes you can ...thats what I do. There is no should when it comes to intramuscular (IM) vs. subcutaneous (SC). Either is fine. There is no local effect nor any real increase in uptake if administered IM. SC was the advantage of being very easy and convenient.
For a lot of human studies they were done via subcutaneous injection.
Never did I see specific reference to intra-muscular. The release curves for the subcutaneous administration looked very similiar to the release curves for I.V. There is not much delay ...a few minutes tops.
Actually unless you are really fat or have scar tissue in the muscle you inject into water based injects have about the same release timing.
I know personally because my insulin injects result in the peak hitting at the same time no matter which way I go.
I also know because after you read study after study you easily see that it doesn't much matter.


6
Eating
You should always try to admininster the CJC/GHRP first and wait a bit before eating.
The reason?
Unlike synthetic GH we are depending on the body to make GH for us. Once GH has been made and is circulating great...but we need to get it made first.
The studies show that circulating fatty acids can really inhibit the production of GHRH and carbs to a lesser extent.
The studies show that circulating fatty acids only blunt but do not inhibit the action of GHRPs. Carbs blunt but to a lesser extent.
It can be argued soundly that it doesn't matter because the CJC (GHRH) is not being made it is being injected. All it needs to do is act on the pituitary...so even in the presence of food it should still function.
However some of GHRP-6s benefit is inducing GHRH release from the hypothalamus. In addition food may blunt some of the pituitary action of GHRH.
So it is best to wait a bit.
How long?
Imagine or refer back to the release curves. GH release happens pretty quickly with either peptide or both together. Within the first 5 minutes the pulse of GH starts to rapidly rise and does so until it peaks at about the 30 minute mark. So I would think that you should always wait at least 10 minutes post administration to eat and if you have the time up to 30 minutes.

In the morning and PWO I administer CJC/GHRP-6 on an empty stomach and wait for 30 minutes. Then my insulin shot (if that happens to be part of my protocol at the time) and I eat.
However pre-bed I usually have plenty of fats in my system. I often (when not dieting) have peanut butter or some combo of fats/protein. I always wait at least 30 minutes after eating my last snack before administering the CJC/GHRP and going to bed.
I don't think the peanut butter in my system really effected my night-time GH release. I've been doing it this way for months and I think it works fine. No night-time hunger and I don't think it interferes with the GH release.
All this to say ...just give yourself a little time between administration & eating. If it is convienent to go 30 minutes do it...if not don't go that long.
Part of the beauty of CJC-1295 is that it stays around. So it is always going to continually act on the pituitary to release GH no matter that you are sometimes eating...
ERRATUM (1/20/2009): Clarification, fats including peanut butter should be avoided well BEFORE administration of peptides. I do not eat fats within a few hours of administration.

7
Storage
Storage
Freeze-dried powder can be stored at -20 degree C for over a year. Reconstituted solution should be kept in the refrigerator at 2-8 degree C and used as early as possible. Avoid multiple freeze-thaw cycles and exposure to frequent for lyophilized powder or reconstituted vials.
Here are a few interesting snippets from studies on storage:

Storage

The following paragraph comes from a study that sent the peptide home with study participants for self-administration.
Vials were stored frozen until dispensed to the subjects, then kept at 4 C for 10 days at home. High performance liquid chromatographic analysis showed that the peptide was stable at 4 C for at least 2 weeks.


The following seems to indicate that GHRP-6 which is a simple peptide chain unlike IGF-1 which is more complex, is relatively resistant to degradation under the right circumstances at room temperature for almost five years.
The influence of the various buffer species (acetate, citrate, phosphate and borate) was shown to be different and the maximum stability of GHRP-6 was revealed to be in acetate buffer of pH 5.5-6.0. Degradation of GHRP-6 was greater in citrate-containing buffers than in acetate-containing ones. Furthermore, in the citrate-containing buffers, the higher buffer concentration caused greater degradation than the lower ones, but the concentration effect was negligible in acetate-containing buffers. Aqueous solution of GHRP-6 buffered with acetate (0.01 M, pH 5.5) showed a predicted t90% of 4.73 years at 20ฐC. - Degradation kinetics of growth hormone-releasing hexapeptide (GHRP-6) in aqueous solution, In Sik Ha… International Journal of Pharmaceutics Volume 144, Issue 1, 22 November 1996, Pages 91-97


It is worth noting the characteristics of the primary aqueous solution we use to reconstitute these peptides:
Bacteriostatic Water for injection, USP is a sterile, nonpyrogenic preparation of water for injection containing 0.9% (9 mg/mL) of benzyl alcohol added as a bacteriostatic preservative. It is supplied in a multiple-dose container from which repeated withdrawals may be made to dilute or dissolve drugs for injection. The pH is 5.7 (4.5 to 7.0)


Or for GHRH
Quote:
Originally Posted by datBtrue;
In general the following applies:
Therapeutic Peptides and Proteins:
Formulation, Processing, and Delivery
Systems, Second Edition
by Ajay K. Banga


5.5.4 Storage in solid state

Lyophilized powders can be quite stable as long as they are not reconstituted.
For example, Activaseฎ lyophilized powder shows no significant loss of
bioactivity after storage for more than 4 years at controlled room temperature.

Degradation in the solid state often takes place by aggregation. The
stability of a protein in the solid state is dependent on the moisture content
of the solid, temperature, and composition of the formulation....


These degradation factors become more important the longer the chain of amino acids becomes & with the inclusion of amino acids sensitive to degradation.

Storage of unreconstituted peptides in frozen form (devoid of moisture) is always far more preferable for long-term storage then reconstituted peptides.

GHRP-6 is a simple chain with no extra-sensitive amino acids. But Growth Hormone Releasing Hormone (GHRH) is more sensitive because it is a longer chain composed of some sensitive amino acids. The added bioconjugation complex of CJC-1295 appears to be stable and shouldn't effect degradation/life of the peptide it is attached to...GHRH.

To answer stability questions the best way is to find studies that specifically examine the stability of the peptide you are interested in.

We have already looked to a very comprehensive study of GHRP-6 now lets look at a stability study for a slightly modified GHRH (and by undeclared assumption CJC-1295).
Investigation of the chemical stability of a new growth hormone-releasing hormone (GHRH) analogue by HPLC, M Idei, I Mezo, EZ Szabo, and G Keri, Biomed Chromatogr, March 1, 1996; 10(2): 89-91

The stability of a new active growth hormone-releasing hormone analogue (D-Ala2,Nle27,(gamma-amino-butyric acid)30-GHRH(1-30)-NH2) was investigated during storage at different temperatures in aqueous solution. Samples stored for various periods of time were analysed by HPLC.

It is concluded that in aqueous solution D-Ala2, Nle27,(gamma-amino-butyric acid)30-growth hormone-releasing hormone (1-30)-NH2 is stable: at least for 36 days at 4 degrees C; for 28 days at 25 degrees C; and for 10 days at 37 degrees C.


GHRP-6, GHRP-2 and Hexarelin
GHRP-6, GHRP-2 and Hexarelin are all interchangeable. They are treated as interchangeable in the studies. They work via the same mode of action. Their slight differences are probably attributable to the different "batches" of non-pituitary neurons they excite. One peptide may excite one "batch" more or less than another.

Hexarelin is the strongest of the GHS peptides. It also induces higher amounts of cortisol & prolactin then the other peptides. It may (according to one comparison study) desensitize quicker. GHRP-2 is a little less strong with less impact on cortisol & prolactin. GHRP-6 has very little impact on cortisol & prolactin (although it is a little elevated above 1mcg/kg dosing) and is a little less stronger than GHRP-2.

So you could choose whichever is cheaper. I know GHRP-6 & GHRP-2 cost the same to make. However GHRP-6 at the moment at retail level is a lot cheaper...

There is no direct benefit to combining GHRPs because they all act through the same mode of action. You just choose one and run it from the saturation dose of 100mcg up to the maximally beneficial dose (which would be 300mcg - 400mcg) at each administration.

I know on the web you see old posts where people talk about the positive effect of combining Hex & GHRP-6. Thats just incorrect. What you do is make a decision on how much GHS you want to run and then choose among the GHS (GHRP-2, GHRP-6, Hexarelin, Ipamorelin). If you really wanted to combine peptides you could choose to use a total peptide dose of say 300mcg which you could apportion half (150mcg) to GHRP-2 and half to GHRP-6 OR use Hexarelin , GHRP-2, GHRP-6 in equal thirds to fill that 300mcg dose slot.

However there is no synergy between these GHRPs and no advantage to dosing these peptides together. I wouldn't combine them to reach my total. I'd just run one and if I ran out and had another on hand I'd continue with that one.

Personally I would be careful with Hexarelin. The upper ranges of dosing 300 - 400 mcg are likely to induce desentsitization and may require time off and will induce the most prolactin & cortisol release.

With GHRP-6 you can dose all the way up to 400mcg & not worry these issues. I don't really worry much about these issues with GHRP-2 either. With GHRP-2 I'd just make sure to avoid dosing much above 200mcg.

CJC-1295 without DAC
Most people are using CJC-1295(without DAC). This is what I call modified GRF(1-29).

GHRH (Growth Hormone Releasing Hormone) as it naturally occurs has 44 amino acids. But because the final 15 amino acids have no effect on GH release (or any other yet ascertained value) they were dropped in the synthetic construction of GHRH. This synthetic construct is called Growth Hormone Releasing Factor (GRF) and the number of amino acids are designated (1-29).

So GRF(1-29) is really the active part of GHRH and it is both a prescription drug called Sermorelin & a research compound called GRF(1-29).

An analog is when changes are made to the structure of GRF(1-29). The primary analogs make these changes by substituting some of the amino acids with others. This is done to increase potency via an increase in receptor-binding strength and or increase half-life by reducing susceptibility to degradation.

The analog that we are most familiar with is modified GRF(1-29), called Tetra-substituted GRF(1-29) in one of the CJC-1295 studies. This analog is also known to us because the Chinese brokers called it this (but is NEVER EVER referred to in scientific literature as) CJC-1295(without DAC).

To the world of science CJC-1295 ONLY refers to GRF(1-29) w/ the modifications, plus a 30th amino acid Lysine attached to a drug affinity complex which enables plasma binding to albumin (post injection).

So to directly answer your question using your terminology CJC-1295(without DAC) is the correct choice.
…………………………………………………………………………………………………………………………………… ………..


ANTI-CATABOLISM -> Ghrelin, GHRP-2, GHRP-6, Hexarelin <- ?
GHRP-2, a GHS-R agonist, directly acts on myocytes to attenuate the dexamethasone-induced expressions of muscle-specific ubiquitin ligases, Atrogin-1 and MuRF1, Daisuke Yamamoto, et al., Life Sciences 82 (2008) 460–466

Introduction

A variety of diseases and conditions, including sepsis, cancer, renal failure, excess of glucocorticoid, denervation and disuse of muscle, can cause muscle atrophy. In these diverse conditions, the atrophying muscles show increased protein degradation through activation of the ubiquitin (Ub)-proteasome pathway (Baracos et al., 1995; Kayali et al., 1987; Price et al., 1996; Tiao et al., 1997; Tischler et al., 1990). It is recently reported that the expressions of Atrogin-1 and MuRF1, both of which are musclespecific Ub-ligases, are involved in protein degradation in muscle and increased in these diverse conditions causing muscle atrophy (Bodine et al., 2001; Gomes et al., 2001; Lecker et al., 2004). Atrogin-1 is a muscle-specific F-box type E3 ligase and reported to be induced 8 to 40 fold in muscle atrophy during fasting, diabetes, cancer and renal failure (Bodine et al., 2001), up to 3 fold in hind limb suspension, immobilization and denervation, and up to 10 fold in cachetic or dexamethasone administration model (Gomes et al., 2001). MuRF1 is a Ring Finger type muscle-specific E3 ligase that is initially found in association with the myofibril (Kandarian and Jackman, 2006) and suggested to play an important role in the myofibrillar proteins breakdown. Both muscle-specific E3 ligases are considered to play a pivotal role in muscle atrophy because knockout mice lacking these E3 ligases are prevented from muscle atrophy (56% sparing for atrogin-1-/- and 36% for MuRF1-/-) (Bodine et al., 2001).

On the other hand, several protective factors for muscle atrophy have been reported. One of the potent protective factors is IGF-I. IGF-I prevents muscle atrophy induced by glucocorticoid (Kanda et al., 1999; Schakman et al., 2005), disuse (Alzghoul et al., 2004) and denervation (Day et al., 2002). IGF-I has a potency to inhibit Atrogin-1 and MuRF1 expressions in atrophying muscle (Sacheck et al., 2004; Stitt et al., 2004). The protective effect of IGF-I for muscle atrophy, at least partly, is exerted by this mechanism (Bodine et al., 2001; Lecker et al., 2004).

Ghrelin stimulates GH release from the pituitary through the GH secretagogue receptor (GHS-R) (Kojima et al., 1999). Also, Growth Hormone Releasing Peptide-2 (GHRP-2), a synthetic ligand for GHS-R, stimulates GH release from the pituitary (Wu et al., 1996). GHRP-2 administration increases plasma GH levels in rats (Sawada et al., 1994) and humans (Pihoker et al., 1995). As a result, plasma IGF-I levels are reported to increase in some studies (Bowers et al., 2004). Thus, GHRP-2 is expected to have a protective action against muscle atrophy via IGF-I. Indeed, a recent report suggested that GHRP-2 was able to prevent arthritis-induced increase in Atrogin-1 and MuRF1 expressions in rat muscle (Granado et al., 2005).

On the other hand, there are reports suggesting the presence of GHS-R in muscle (Papotti et al., 2000; Pierno et al., 2003) and the signal transduction mechanism of ghrelin is partly similar to those of IGF-I and insulin (Murata et al., 2002). Hence GHS-R ligands may play a role in the process of muscle atrophy.

In the present study, we have examined the effect of GHRP-2 on Atrogin-1 and MuRF1 mRNA levels in dexamethasoneinduced muscle atrophy in the rats, as a model of muscle atrophy that is often observed during steroid hormone-treatment in human. We have further tested whether the effect is a direct action on myocytes through GHS-R and found for the first time that GHRP-2 directly acted on myocytes.

...

Discussion

In the present experiment, we found that GHRP-2 attenuated Atrogin-1 mRNA level induced by dexamethasone in ratmuscles. Although the mechanism by which dexamethasone causes muscle atrophy is unknown, one possibility is via enhancement of glutamine synthetase activity (Falduto et al., 1992a,b) and the other is via induction of Atrogin-1 expression (Bodine et al., 2001; Lecker et al., 2004).

GHRP-2 has an action to stimulate GH secretion from pituitary, which in turn could increase plasma IGF-I levels. Since IGF-I has been reported to be a growth factor causing muscle hypertrophy (Kanda et al., 1999; Schakman et al., 2005), the elevation of plasma IGF-I levels may affect dexamethasoneinduced muscle atrophy. Interestingly IGF-I has been already reported to attenuate Atrogin-1 expression in vivo (Sacheck et al., 2004; Stitt et al., 2004). In the present study, however, plasma IGF-I levels were not changed by the treatment with GHRP-2. This finding was consistent with previous reports that GHRP-2 did not increase plasma IGF-I levels in mice (Tschop et al., 2002) and humans (Nijland et al., 1998), suggesting that GHRP-2 does not always increase plasma IGF-I levels. Our data rather suggested that the reduced mRNA levels of Atrogin-1 and MuRF1 in muscle by GHRP-2 was not due to the rise of circulating IGF-I levels. In addition, IGF-I expression in soleus muscles was not affected by GHRP-2 in the present study. Recently, Granado et al. (2005) reported that subcutaneous daily administration of GHRP-2 (100 ug/kg) decreased expression of Atrogin-1 and MuRF1 in atrophic muscle of adjuvant-induced arthritis rats. In their report, plasma IGF-I level was much lower in arthritis rats than in normal control and GHRP-2 did not increase muscle IGF-I mRNA level. Their findings, consistent with our findings, suggested that GHRP-2 decreased Atrogin-1 and MuRF1 mRNA levels through a pathway other than circulating IGF-I and local IGF-I production.

Binding assay using GHS-R ligands has shown specific binding sites in muscle (Papotti et al., 2000) and in vitro application of ghrelin or ghrelin agonists modulated chloride and potassium conductance in rat muscle (Pierno et al., 2003). These findings suggest the presence of GHS-R in skeletal muscle. In this experiment, we found the expression of GHSR-1a in differentiated C2C12 cells. We have already reported that intracellular signal pathways of ghrelin were partly similar to those of insulin and IGF-I (Murata et al., 2002). From the above reasons, we speculated GHRP-2 might work in myocytes to suppress Atrogin-1 and MuRF1 mRNA levels like IGF-I and examined whether GHRP-2 has a direct action on myocytes to inhibit Atrogin-1 and MuRF1 mRNA expressions. GHRP-2 dose-dependently suppressed dexamethasone-induced Atrogin- 1 and MuRF1 expressions in C2C12 cells. These findings indicate that GHRP-2 directly acts on myocytes and attenuates the level of Atrogin-1 and MuRF1 mRNA.

To further clarify a direct suppressive effect of GHRP-2 on Atrogin-1 and MuRF1 mRNA levels, [D-Lys3]-GHRP-6, a GHS-R-1a antagonist was used in C2C12 cells. There are two types of ghrelin receptors, GHS-R-1a and GHS-R-1b (Howard et al., 1996; Mckee et al., 1997). GHS-R-1a is an active receptor mediating ghrelin action. GHS-R-1b, a splicing variant of GHSR-1a, does not mediate ghrelin signal. We examined the specificity of GHRP-2 action using [D-Lys3]-GHRP-6. We found that [D-Lys3]-GHRP-6 partly and completely reversed the suppressive effects of GHRP-2 on Atrogin-1 and MuRF1 mRNA levels, respectively. These results suggest that GHRP-2 directly inhibits Atrogin-1 and MuRF1 mRNA level through GHS-R-1a.

Since C2C12 cells produce IGF-I (Frost et al., 2003), paracrine or autocrine action of IGF-I may be involved in the suppressive effect of GHRP-2 on Atrogin-1 and MuRF1 mRNA level. To elucidate this possibility, we measured IGF-I mRNA level in C2C12 cells. However, we were not able to find the increase in IGF-I mRNA in C2C12 cells in response to GHRP-2, suggesting that locally produced-IGF-I in C2C12 cells is not involved in the suppressive effect of GHRP-2 on Atrogin-1 and MuRF1 mRNA levels. Dexamethasone also did not influence IGF-I mRNA level in C2C12 cells, although it decreased IGF-I mRNA level in vivo soleus muscle. These results suggest that dexamethasone has an indirect action to reduce IGF-I mRNA level in muscles in in vivo animals. Glucocorticoid is reported to inhibit pulsatile GH secretion (Giustina and Veldhuis, 1998) and reduce GH receptor expression (King and Carter-Su, 1995). As a result, IGF-I mRNA level was thought to decrease in vivo experiment in the present study. Dexamethasone has been reported to reduce the expression in in vivo animals (Gilson et al., 2007), being consistent with our in vivo result.

In summary, GHRP-2 suppressed dexamethasone-induced Atrogin-1 mRNA expressions in in vivo rats without elevating plasma IGF-I and IGF-I mRNA in muscle. Furthermore GHRP-2 decreased dexamethasone-induced Atrogin-1 and MuRF1 expressions in C2C12 myocytes. This effect was blocked by the addition of [D-Lys3]-GHRP-6, a GHS-R-1a antagonist. These findings suggest that a direct action of GHRP-2 through GHSR-1a suppresses Atrogin-1 and MuRF1 mRNA levels in C2C12 cells. GHRP-2 might lead to the protection of muscle atrophy induced by dexamethasone.
…………………………………………………………………………………………………………………………………… ……………………..
ANABOLISM -> Ghrelin, Des-Acyl Ghrelin, GHRP-2, GHRP-6, Hexarelin = Promote Differention & Fusion of Muscle Cells <- ?
Ghrelin and Des-Acyl Ghrelin Promote Differentiation and Fusion of C2C12 Skeletal Muscle Cells, Nicoletta Filigheddu, Molecular Biology of the Cell Vol. 18, 986–994, March 2007

...

DISCUSSION

Skeletal muscle satellite cells are quiescent mononucleated myoblasts, located between the sarcolemma and the basal membrane of terminally differentiated adult muscle fibers. On muscle diseases or direct injury, quiescent satellite cells are activated to undergo proliferation and eventually differentiate to allow muscle regeneration.

Skeletal muscle regeneration involves, sequentially, satellite cell proliferation, commitment to terminal differentiation, cell fusion into multinucleated syncitia, and muscle fiber formation.

Such mechanisms leading to muscle regeneration are poorly understood; they seem to recapitulate the embryonic program of differentiation, although the extracellular factors regulating such processes may be different.

Satellite cell differentiation into skeletal muscle can be subdivided into temporally separable events, coordinated by the expression of proteins of the muscle regulatory factors family, such as myogenin, and of cyclin-dependent kinase inhibitor of the p21 family (Andres and Walsh, 1996), resulting in cell cycle exit and commitment to terminal differentiation. Later on, expression of muscle contractile proteins, such as MHCs and myosin light chains (MLCs), are hallmarks of phenotypic differentiation. Finally, fusion of myocytes into multinucleated myotubes is the terminal step of muscle differentiation.

The growing interest in skeletal muscle regeneration is associated to the opening of new therapeutic strategies for several muscular degenerative pathologies such as dystrophies, muscular atrophy, and cachexia associated to aging, cancer, chronic heart failure, and acquired immunodeficiency syndrome as well as the treatments of skeletal muscle injury after trauma.

Although Ghrelin (GHR) is a circulating hormone mainly secreted by the stomach, it is also synthesized in a number of tissues, suggesting both endocrine and paracrine effects (Gnanapavan et al., 2002).

The evidence that 1) Ghrelin (GHR) up-regulation is specifically associated to either congestive heart failure (CHF)- or cancer- induced cachexia (Nagaya et al., 2001, Shimizu et al., 2003) and that its administration strongly prevents CHF associated cachexia (Nagaya et al., 2004); 2) GHR, (Des-Acyl Ghrelin) D-GHR, and Growth Hormone Secretagogues (GHSs) inhibit apoptosis of cardiac myocytes (Filigheddu et al., 2001; Baldanzi et al., 2002); and 3) skeletal muscle features high binding sites for synthetic GHSs (Papotti et al., 2000), lead us to speculate that GHR and D-GHR may act directly also on skeletal muscle. Indeed, we observed that both GHR and D-GHR stimulate tyrosine phosphorylation of several proteins and activate ERK-1/2 and Akt (data not shown), indicating that both factors could exert a biological activity on these cells.

Here, we show that nanomolar concentrations of both GHR and D-GHR induce the differentiation of proliferating skeletal myoblasts in a concentration-dependent manner and promote their fusion into multinucleated syncitia in vitro. The cellular and molecular mechanisms by which GHR and D-GHR elicit these responses are not known. Cell cycle withdrawal is a prerequisite for myogenic terminal differentiation (Walsh and Perlman, 1997). Indeed, the ability of GHR and D-GHR to reduce DNA synthesis of proliferating C2C12 myoblasts is highly consistent with their prodifferentiative activity. However, inhibition of cell proliferation is not sufficient to elicit muscle differentiation. For example, myostatin inhibits both proliferation and differentiation of C2C12 myoblasts, through down-regulation of MyoD and myogenin expression (Joulia et al., 2003). Conversely, GHR and D-GHR, beyond inhibiting cell proliferation, induce the expression of myogenin, which is required for the complete program of differentiation of skeletal myoblasts to proceed (Zhang et al., 1999). To our knowledge this is the first evidence for an extracellular factor able to induce muscle differentiation of proliferating skeletal myoblasts in GM.

In proliferating C2C12 myoblasts, activation of p38 pathway obtained by overexpression of constitutively active MKK6 is sufficient to induce myogenin expression, cell cycle exit, and skeletal muscle terminal differentiation (Wu et al., 2000). Thus, we investigated whether GHR and D-GHR prodifferentiative activity is mediated by p38. Consistently, inhibition of p38 by cell treatment with SB203580 resulted in the partial albeit significant inhibition of GHR and D-GHR induced differentiative activity. In addition, we also showed that both GHR and D-GHR activate p38. Altogether, these data demonstrate that GHR and D-GHR act as antiproliferative and prodifferentiative factors by stimulating the p38 pathway.

The lack of expression of Growth Hormone Secretagogue Receptor One-A (GHSR-1a) in either C2C12 myoblasts and skeletal muscle tissue (Gnanapavan et al., 2002) as well as the activity exerted by D-GHR suggest that GHR and D-GHR–differentiating activities are mediated by a yet unidentified receptor, common to both acylated and unacylated peptide and distinct from GHSR-1a. Indeed, here we showed that C2C12 cells feature high-affinity common binding sites for both GHR and D-GHR. Such binding sites are specific, because they do not recognize either N-terminal truncated ghrelin or motilin, which are unable to induce differentiation. These studies also demonstrate that the N-terminal portion of the GHR peptide is required for binding and induction of C2C12 muscular differentiation. Together, these data provide further evidence for novel GHR receptor subtypes, which do not discriminate between the acylated and unacylated peptide. Although evidence for common GHR and D-GHR receptors have been reported in several cells, including a cardiomyocyte-derived cell line (Baldanzi et al., 2002), this is the first evidence for their expression in skeletal muscle.

We also verified whether the ghrelin gene is up-regulated in C2C12 myoblasts induced to differentiate in DM. However, no difference of ghrelin expression was detected by real-time RT-PCR between proliferating and differentiating cells (data not shown), suggesting that GHR gene product is not involved in DM-induced skeletal muscle differentiation in vitro.

By showing that GHR and D-GHR stimulate terminal differentiation of skeletal myoblasts in vitro, we may raise the hypothesis that the function of GHR gene may be involved in skeletal muscle differentiation in vivo. However, the lack of a consistent phenotype in GHR knockout mice, suggests that GHR function is not required for myogenesis during development. Consistently, we have not detected any GHR expression in somites or related structures during embryonic development by in situ hybridization (data not shown). However, although not essential for embryo development, GHR might be involved in the complex process of myogenesis in the adulthood, i.e., in regenerative processes of skeletal muscle. This hypothesis is consistent with the data showing that FGF6 is not required for muscle development, but is required in the adult for damage-induced muscle regeneration (Floss et al., 1997).

Upon muscular injury, skeletal myoblasts are activated to terminally differentiate through an autocrine/paracrine loop. We may speculate that GHR would contribute to skeletal muscle plasticity, promoting the differentiation and fusion of myoblasts in the damaged muscles. If this hypothesis would be proved, the activation of the receptor mediating GHR and D-GHR differentiative activity as well as the over-expression of the hormone may provide novel therapeutic strategies for the reduction or retardation of several skeletalmuscle pathologies, including dystrophies, atrophies, and cachexia.
…………………………………………………………………………………………………………………………………… ………………..
Mixed Peptides
Storage: Reconstituted (mixed) vials must be stored in the refrigerator (2 to 8 degrees celcius).
Expiry: Use within 8 weeks (2 months) of mixing.
It is NOT recommended to pre-load syringes and freeze them. When the peptide freezes there is a risk that it may come out of solution into particles. When it is thawed out again it may not return into solution, meaning particles will be injected which can cause irritation or damage at the injection site.
Note: PRE mixed peptide vials should not be shaken under any circumstances. None of our peptides come pre mixed and ALL packages include free solvent.
Unmixed Peptides
Storage: In powder (lyophilized) form vials should be stored frozen (0 to -20 degrees celcius).
Expiry: Will remain stable up to 48 months (4 years) in the freezer.
Unmixed vials can be stored in the freezer for a period of up to 48 months (4 years); however, if you are going to use the vials within 1 months then we recommend simply storing them in the refrigerator as repeated freeze-thaw cycles can cause damage to the peptide.
Storage Safety
Both mixed and unmixed vials should be stored inside sealed plastic satchels or containers and kept away from food in the refrigerator or freezer.
Also
CJC1295 without dac IS THE SAME AS modified GRF (1-29)...
Modified CJC or Dac-less CJC is just TSubed GRF 1-29(4 amino substitutions) . Halflife is about a couple of hours and needs to be dosed multiple times per day.
The health benefits of GHRP-2
The health benefits of GHRP-2 are immense, and it has enormous applications in the field of age management as practiced by centers like the Age Management Panama clinic. A normal part of the aging process for many people is a decrease in a variety of hormone levels, and an important part of the anti aging therapy process is to asses any existing hormone deficiencies. These deficiencies can be treated with GH Releasing Peptide-2, either in isolation or with other hormone therapies in order to have astounding effects.
Side effects:
The major side effect accompanied by the use of GHRP-2 or other GH secretagogues is a significant increase in appetite because secretagogues mimic the action of Ghrelin, a peptide which is released naturally in the lining of the stomach and increases hunger and gastric emptying.
More information:
It has also been discovered that when GHRP-6 and insulin are used simultaneously, GH response to GHRP-6 is increased. A recent study in normal mice showed significant differences in body composition, muscle growth, glucose metabolism, memory and cardiac function in the mice being administered the GHRP-6. There are still many questions regarding this fairly new compound, scientists are hoping to gain a better clinical understanding of the peptide through further research over the next few years.

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Old 06-07-2010, 11:41 AM
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Nice Rep-keep us updated on how this works-read so many conflicting stories on this
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Old 06-07-2010, 04:00 PM
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nice read, pretty imformative
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Old 06-07-2010, 10:33 PM
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Nice Rep-keep us updated on how this works-read so many conflicting stories on this
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nice read, pretty imformative
bump & bump, definately let us know how this goes for you REP.....am curious about this in the future........
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Old 06-08-2010, 06:41 AM
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Please let me know how you get on with this as defiantly what to try some,
Only thing holding me back @ the moment is the price down in the south,
If you don't mind me asking what sort of money for a course,
As pricing being offered to us, may be a rip off.
Don’t mind paying if it's fare lol
I have a friend, who also is interested as we are getting a bit older
Thanks

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Old 06-08-2010, 01:40 PM
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P.M'd ye Jimbo
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Old 06-18-2010, 10:40 AM
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goods came on Wednesday 16/06 and for any one interested
I got gear from www.propeptides.net whom supply the bac water free with the goods .........awesome service too.
so started on Wed 16/06.
One thing tho,you can only get 4ml of bac water in teh vial, they are smaller than I thought.
My first thoughts were to put 5ml in the GHRP-2 vial as it holds 5mg which for easy coversion would have been 100 mcg per ml.
So this vial will have to be 160IU's to get the 200mcg of GHRP-2.
Lesson learned here tho next vial I will only add 2.5ml of Bac water and will only need 100IU to get the 200 mcg and keep it simple



First Injection was fine no upset stomach that Ive read about. I was a little hotter than normal through-out the night, but not sure if this was the stuff or the weather. did wake up a few times more but this could have been the hotter weather too ..dont thing you can really go off ist inject any way tho.
Will report back after a week.
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Old 06-18-2010, 10:55 AM
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I know the GHRP-6 I used was instant effect. never used CJC 1295 before, so have no first hand info on that compound.
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Old 06-19-2010, 04:35 AM
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I know the GHRP-6 I used was instant effect. never used CJC 1295 before, so have no first hand info on that compound.
WW,can you elaborate on what you mean by instant effect please.
I have done 4 injects now and i definetly notice an extra warmth in bed with it and I also waken up 3 -4 more times as well although I can get back to sleep again no problem
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Old 06-25-2010, 03:44 PM
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well thats just over a week now and not much to report at this point
Still get the heat at night but sleeping a bit better tho
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Old 06-25-2010, 04:19 PM
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There is just to many numbers,I had to pull my socks off to figure that out.

Then I found out it was in the TITLE.

I so underachieved ,Im going to go put more ass cream on,so depressed, wait thats the crack.
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Old 06-30-2010, 10:39 AM
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well thats a couple of weeks tonight now and not much to report at this point
Still get the heat at night but sleeping totally okay now.
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Old 06-30-2010, 02:45 PM
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Keep us informed REP will be really interesting to see how u go. Are you using anything else with it ? or just GHRP-2 CJC-1295
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Old 06-30-2010, 02:58 PM
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using it in addition to the cycle I am on at the moment which is
600mg Deca per wk and 400mg test per week (cypionate 200mg enathate 200mg)
And hey Badstone if you ever visit this thread you are right this combo does work well. and I always thought the TEST had to be higher ..what rot.
So thanks for the heads up on that one.


But this will finish in 2 weeks an then I will be back to a TRT dose for 8 weeks which this time will be 300mg sust
After all these years I know now what the gear does to me so aynthing extra I wll pick up on no bother.
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Old 06-30-2010, 03:22 PM
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using it in addition to the cycle I am on at the moment which is
600mg Deca per wk and 400mg test per week (cypionate 200mg enathate 200mg)
And hey Badstone if you ever visit this thread you are right this combo does work well. and I always thought the TEST had to be higher ..what rot.
So thanks for the heads up on that one.


But this will finish in 2 weeks an then I will be back to a TRT dose for 8 weeks which this time will be 300mg sust
After all these years I know now what the gear does to me so aynthing extra I wll pick up on no bother.
REP
Oh Yeah I like mixing and matching test. Sometimes I go up to 500mg but most of the time 400mg does the trick.... Once you learn how your body works to what compound you can make it happen.
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Old 06-30-2010, 03:41 PM
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And hey Badstone if you ever visit this thread you are right this combo does work well. and I always thought the TEST had to be higher ..what rot.
So thanks for the heads up on that one.
REP
Good thread. will be keeping an eye on this.

Yeah..not sure when the test had to be higher methodology came into play. Certainly was not a rule in the 80's. Glad to see people looking outside the box again.
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Old 06-30-2010, 05:23 PM
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deffinately keep us updated on this, rep. Im planning a peptide only cycle after my shic. I m gonna try hexorelin, semorelin and cjc1295. Im interested in what kind of results you get
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Old 07-02-2010, 03:39 PM
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okay another wee update as it happens.

appetite has slowly went up a notch but having to control that and not eat any more than I already am, or this would scupper the BF% thing Im keeping an eye on. Definelty attribute this to the Peps.
Rep

Ps) And by the way Badstone ......... I imagine with the test being at 400mg a week... no deca dick all is fine in this area. thank F**k

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Old 07-03-2010, 07:56 AM
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okay another wee update as it happens.

appetite has slowly went up a notch but having to control that and not eat any more than I already am, or this would scupper the BF% thing Im keeping an eye on. Definelty attribute this to the Peps.
Rep

Ps) And by the way Badstone ......... I imagine with the test being at 400mg a week... no deca dick all is fine in this area. thank F**k

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If you do start to feel a little deca dick just up the test a little until you find that magic number that works for your body. 400mg is the magic number that worked for me. I started at 200mg and went up from there.
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Old 07-03-2010, 09:26 AM
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If you do start to feel a little deca dick just up the test a little until you find that magic number that works for your body. 400mg is the magic number that worked for me. I started at 200mg and went up from there.
I ve always loved decca. I have always ran it w/ a test base and have never experienced problems w/ performance. Quite the opposite.
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Old 08-21-2010, 04:34 AM
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Right , thats me been on this stuff for just over 2 months now so time for an update.
I do believe it has helped me along with the gear increase my poundages just that litte bit more on cycle.
I dont really suffer from acne but I get a few spots here and there, but have found that they are even less on this stuff , my skin also feels ..........well............ smoother...in a better condition. it also has helped in its elasticity ..but not by much ..............who knows how goods it will be six months in.
now here's the unexpected thing , my joints are not so creaky (knees especially) since introducing the peptides, they feel a lot easier too
Thats it for now
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Old 08-21-2010, 06:14 AM
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Yeah I notice a big difference in joint comfort while on GH. It does lubricate dried out old guy joints and I think this softens the connective tissue just a tad allowing them to be more flexible. Never tried those other peps tho but I imagine they all do basically the same.
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Old 08-21-2010, 08:00 AM
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Yeah I notice a big difference in joint comfort while on GH. It does lubricate dried out old guy joints and I think this softens the connective tissue just a tad allowing them to be more flexible. Never tried those other peps tho but I imagine they all do basically the same.
yea ..well Im 55 now so need all the lube I can get
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Old 08-21-2010, 08:38 AM
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WW,can you elaborate on what you mean by instant effect please.
I have done 4 injects now and i definetly notice an extra warmth in bed with it and I also waken up 3 -4 more times as well although I can get back to sleep again no problem
REP
sorry for the late reply. I was taking 600mcg right after working out and GHRP-6 made me want to pig out which is great if your wanting to pack on muscle. just have the right foods around when you take the shot, so you don't pig out on the wrong foods and get fat.
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Old 08-22-2010, 11:53 AM
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sorry for the late reply. I was taking 600mcg right after working out and GHRP-6 made me want to pig out which is great if your wanting to pack on muscle. just have the right foods around when you take the shot, so you don't pig out on the wrong foods and get fat.
Bloody hell WW thats high LOL ...I should know you by now tho m8 .you dont mess with piddly numbers LOL.
yea I always have the right foods around ............more eggs and porridge .............. Ive never had to get up and eat at 2am before I introduced the peps.
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