The results of the present study indicate that NO-Shotgun® and NO-Synthesize® supplementation provided before and after resistance exercise, respectively, and in conjunction with heavy resistance training, is more effective than carbohydrate at increasing fat-free mass, muscle strength and mass, and markers of muscle protein synthesis in untrained males, while having no effect on whole blood and serum clinical safety markers. In regard to the various ingredients contained in both supplements, based on previous research it is conceivable that the primary active ingredients are whey protein, creatine, leucine, beta-alanine, and KIC.
Our rationale to use carbohydrate as a comparator was based on the premise that there is empirical evidence to suggest that carbohydrate supplementation prior to and after resistance exercise results in the maintenance of muscle glycogen , in addition to the fact that the insulin response associated with carbohydrate ingestion up-regulates signal transduction pathways in muscle which can activate muscle-specific gene expression and protein synthesis . Therefore, many recent studies have provided nutrient provision in close proximity (either before and/or after) to resistance exercise and, in so doing, have used carbohydrate as a comparator based on the premise that carbohydrate provided in conjunction with other nutrients such as protein , amino acids , and creatine  has been shown to augment the responses to resistance training.
Our results demonstrated that both NOSS and CARB significantly increased total body mass (p = 0.001) with no associated increases in total body water (p = 0.345). Additionally, fat-free mass was increased in both groups (p = 0.001) with NOSS demonstrating significantly greater improvements than CARB (p = 0.023). These findings for NOSS are similar to our previous study  as well as a study that observed 12 weeks of heavy resistance training and creatine supplementation to induce a greater increase in fat-free mass compared to the carbohydrate group . In addition, 10 weeks of heavy resistance training and whey protein and amino acid supplementation resulted in greater increases in fat-free mass compared to carbohydrate .
Increases in both upper- (p = 0.023) and lower-body (p = 0.035) muscle strength were significantly greater in NOSS compared to CARB. The present data are corroborated by our previous study , along with previous other studies which have demonstrated heavy resistance training, when combined with creatine [9, 19] whey and casein protein and amino acids , and whey protein and amino acids  produces greater increases in muscle strength.
Our present results demonstrated that NOSS supplementation results in preferential increases in myofibrillar (p = 0.049) protein and Type I (p = 0.013) and IIa MHC (p = 0.046) when compared to carbohydrate. Our results are similar to a study in which creatine supplementation in conjunction with 12 weeks of resistance training resulted in an increase in myofibrillar protein and MHC isoform content when compared to carbohydrate . Additionally, a protein and amino acid supplement ingested in concert with 10 weeks of heavy resistance training induced a greater increase in myofibrillar protein compared to carbohydrate .
As with our previous study , we observed serum IGF-1 to be increased with heavy resistance training after four weeks; however, there was no difference between groups (p = 0.385). Previous studies have demonstrated heavy resistance training to either increase  or have no effect  on serum IGF-1. We have previously shown that 10 weeks of heavy resistance training combined with a daily supplement containing whey/casein protein and free amino acids increased circulating IGF-1 levels . Even though serum IGF-1 was increased in the present study, we can conceivably conclude that none of the ingredients contained in the supplements ingested by both groups served as IGF-1 secretagogues. Even so, this outcome may not be germane to the results as hepatically-derived circulating IGF-1 appears to have no direct effect on muscle hypertrophy  compared to skeletal-muscle derived IGF-I which has been shown to increase in response to resistance training  and induces muscular protein synthesis [24, 25].
The MRFs (Myo-D, myogenin, MRF-4, myf5) are transcription factors that play a role in muscle hypertrophy by binding to E-boxes in the promoter region of various sarcomeric genes , thereby up-regulating transcription which can invariably lead to an increase in protein synthesis. It appears that myogenin and MRF-4 specifically up-regulate the expression of genes specific to contractile protein [26, 27] and fast and slow muscle fiber differentiation . Type I and II muscle fibers have been shown to preferentially accumulate myogenin and Myo-D, respectively . Creatine supplementation in conjunction with resistance training has been shown to increase MyoD, myogenin, and MRF-4 that were correlated with increased MHC and myofibrillar protein  and myofiber size . In line with our previous studies, the present results demonstrated that both groups underwent significant increases in MRF content [15, 29] and all three MHCs [15, 19]. However, NOSS underwent even greater increases in Myo-D (p = 0.038) and MRF-4 (p = 0.001) and Type 1 and 2A MHC. It is difficult to conclude which specific ingredient elicited these results; however, based on previous research we can speculate a role for creatine since 12 weeks of supplementation and heavy resistance training resulted in muscle hypertrophy, along with concomitant increases in MHC Type 1, 2A, and 2X protein, and myofibrillar protein .
As with most nutritional supplements, NO-Shotgun® and NO-Synthesize® are comprised of a number of different compounds with most having no little, if any, clinical safety data available. During the course of the study, we observed no significant changes beyond the normal clinical ranges in regard to clinical safety measures in either group. These data indicate that the ingestion of carbohydrate, NO-Shotgun®, and NO-Synthesize® for a period of 28 days has no detrimental clinical effects with regard to the whole blood and serum variables assessed.
A purpose of the present study was to compare the effects of NO-Shotgun® given pre-exercise and NO-Synthesize® given post-exercise to our previous study (15) in which only NO-Shotgun® was given pre-exercise to determine if additional post-exercise nutrient provision would provide an augmented effect. Our present data showed that total body mass to increase by 1.51% and fat-free mass to increase 3.66% in the NOSS group. This mirrors the results observed in our previous study in which the NO-Shotgun® (NO) group increased 2.59% and fat-free mass 4.75% (15). In our previous study, upper- and lower-body strength increased 8.82% and 18.40%, respectively, with only lower-body being greater than placebo (15). However, our present results show a preferential increase of 12.62% and 21.28%, respectively, for upper- and lower-body strength with both being greater in NOSS. In our previous (15) and present study, serum IGF-1 increased 9.34% and 3.38%, respectively, with neither being different from placebo. Myofibrillar protein was preferentially increased by 70.39% in the NO group in our previous study (15) and 29.21% in our present study. For the MRFs, in our previous study, they were preferentially increased in the NO group by 70.91%, 56.24%, and 71.17% for Myo-D, MRF-4, and myogenin, respectively. While still preferentially increased in the NOSS group in our present study, Myo-D, MRF-4, and myogenin increased 18.02%, 20.32%, and 13.49%.
NO-Shotgun® and NO-Synthesize® contain a proprietary blend of a number of compounds assumed to be effective at increasing muscle strength and mass such as creatine, arginine, glutamine, beta-alanine, keto-isocaproate (KIC), and leucine, casein and whey protein, branched-chain amino acids, lysine, phenylalanine, threonine, histidine, and methionine. As a result, attempting to isolate which specific ingredient has the greatest impact on our outcome measures is not feasible. However, our present results indicate that supplementation protocol of providing NO-Shotgun® pre-exercise, NO-Synthesize® post-exercise, and NO-Synthesize® on non-exercise days for 28 days is more effective than carbohydrate at increasing muscle mass and strength and markers indicative of muscle protein synthesis, while having no negative impact on the clinical chemistry variables assessed. Furthermore, our present results demonstrate preferential improvements in muscle strength and mass and agree with our previous study , and suggest that nutrient provision before and after resistance exercise is effective in preferentially augmenting muscle strength and mass.