The

lobulation of the fetal liver begin near the liver hilum at the 9th WD, and progresses from the hilum to the periphery of the liver until at about 1-month post partum. Concerning the future lobular area, HSC and the second layer cells around the centrolobular veins, derive from mesenchymal cells, as well as the mesenchymal vessels which formed the primitive hepatic sinusoids [9, 10]. Concerning the portal tract, its centrifugal development is closely associated with intra-hepatic biliary tree development [11]. Depending exclusively on the location of the portal tract along the portal tract tree, between the hilum and the periphery, the sequence of maturation of a portal tract schematically comprises 3 stages [12]: 1) At the ductal plate stage, this website segments of double-layered cylindrical or tubular structures, called ductal plate, outlined Selleck KU 57788 the future portal tract. The future portal tract contains also large portal vein branch and limited stroma; 2) At the ductal plate remodelling stage, the tubular structures become incorporated into the stroma surrounding the portal vein branch and the rest of the ductal plate involutes. Arterial branches are also present; 3) At the remodelled stage, the portal tract is mature: it contains a branch of the portal vein, two branches of the hepatic artery

and two bile ducts [13]. In cases of ductal plate malformation, notably observed in Ivemark’s renal-hepatic-pancreatic dysplasia or Ivemark’s dysplasia syndrome type II (IDS2), in

Meckel-Gruber syndrome (MKS) and in autosomal recessive Epigenetics inhibitor polycystic kidney MLN2238 disease (ARPKD), the portal tract was deeply modified [14–16]. It was characterised by portal tract fibrosis, more mesenchymal cells with ASMA expression and increased number of arteries [11, 17]. The aims of our study were to follow principally the ASMA, h-caldesmon, CRBP-1 expression of mesenchymal cells during the normal development of the fetal liver and to explore the phenotypic evolution of the portal tract mesenchymal cells during the abnormal development of fetal liver presenting fibrosis following ductal plate malformation. Results Normal fetal liver – Histology In all tissue samples, the fetal liver tissues showed anastomosing sheets of fetal hepatocytes. Each sheet, being two or several cells in thickness, was separated from the others by capillaries. Haematopoiesis was present in all cases and prominent in the capillary lumen or in the Disse space after 12 WD. After 11 WD, future portal tracts appeared in the parenchyma and developed with a centrifugal manner from the hilum to the periphery of the liver. Depending on the tissue section level (near the hilum or at the periphery), the 3 portal tract maturation stages (described above) were present. In the parenchyma, future centrolobular veins with a thin wall were present.

MAC participated in the design of the study, interpretation of data and helped to draft the manuscript. CZA performed the PCR screenings and helped in the laboratory work.

MBZ provided Trichostatin A ic50 the strains and drafted the manuscript. EC participated in the conception of the study, the interpretation of the data and helped to draft the manuscript. CS participated in the design of the study, performed part of the laboratory work, interpreted the data and drafted the manuscript. All authors read and approved the final manuscript.”
“Background Regulated promoters are commonly used in recombinant Lazertinib protein production processes and are particularly important for production of host-toxic proteins or proteins that cause a serious metabolic burden to the host cells [1, 2]. The transcription regulator XylS stimulates expression from the Pm promoter in the presence of benzoic acid Chk inhibitor or derivatives thereof [3]. XylS originates from the Pseudomonas putida TOL-plasmid and is expressed from two different promoters, Ps1 and Ps2: Ps1 is regulated,

while Ps2 is constitutive [4]. The production level of XylS from Ps2 is low, leading to an estimated amount of about 200 molecules per cell [5]. XylS belongs to the AraC/XylS family of transcription factors and it has been shown to be transcriptionally active as a dimer. Dimerization occurs both in the absence and presence of inducer, but to a greater extent in its presence [5, 6]. In spite of sequence similarities and common functional domains, the

different members of the AraC/XylS family act via a range of different mechanisms. AraC, for example, forms dimers like XylS, both in the presence and absence of inducer [7]. In the presence of inducer Avelestat (AZD9668) it acts as an activator of gene expression (like XylS), but in the absence of inducer, it represses gene expression via DNA bending. The first two proteins of the AraC/XylS family, for which 3D crystal structures have been determined, were RobA and MarA, and both exist as monomers only [8]. XylS consists of two domains and structural models exist for both, constructed based on sequence alignments [9, 10]. The model of the N-terminal domain proposes a β-barrel, which is involved in inducer binding and two α-helices that probably are involved in dimerization [10–12]. In the C-terminal domain seven α-helices that form two helix-turn-helix motifs are proposed [9]. These motifs are responsible for binding to two direct repeats with the sequence TGCAN6GGNTA upstream of the -35 box of Pm[13, 14]. The second binding site overlaps by two bases with the -35 box and this overlap is essential for transcription initiation from Pm[15]. Both domains are thought to interact with the host RNA polymerase (RNAP) [16–19]. The N-terminal domain has been shown to suppress the action of the C-terminal domain in the absence of inducer [5, 20]. Binding of wild type XylS to DNA can only be observed when the protein is dimerized [5].

The results indicate that both bio- and chem-AuNPs are largely ineffective at inducing ROS generation in MDA-MB-231 cells, whereas H2O2- and AgNP-treated groups showed remarkable increase in ROS generation (Figure  9).

Figure 9 The effect of AuNPs in ROS generation. Relative fluorescence of DCF was measured using a spectrofluorometer with excitation at 485 nm and emission at 530 nm. The results are expressed as the mean ± SD of three separate experiments, each of which contained three replicates. Treated groups with bio- and chem-AuNPs were not find more statistically different from the control group based on the Student’s t test. (p > 0.05). H2O2- and AgNP-treated groups were statistically buy CB-839 different from the control group based on the Student’s t test (*p < 0.05). Chuang et al. [71] extensively GDC-0973 in vitro studied the exposure of three different-sized AuNPs in human gastric carcinoma (AGS) and human lung adenocarcinoma epithelial (A549) cells. Their results suggest that significant

increases of ROS generation occur with certain concentrations of AuNPs in AGS cells. Conversely, no obvious increases were observed for A549 cells in any of the three sizes of AuNPs. The authors eventually concluded that ROS signaling may play a role in AuNP-induced apoptotic cell death in AGS cells. Furthermore, western blot analyses revealed that the expression of proteins involved in the anti-oxidative defense system was not significantly modulated any of the three sizes of AuNPs in both lines, except for a modest increase in TrxR-1 and SOD-1 in AGS cells [71]. Altogether, our results

suggest that biologically synthesized very AuNPs have significant biocompatibility and could possibly be used for ultrasensitive detection, gene transfer, biomolecular imaging, drug delivery, and cancer therapy. Conclusion Synthesis of nanoparticles using biological systems is an important area of nanobiotechnology. Here we show a simple, rapid, clean, efficient, cost-effective, and green method for the synthesis of biocompatible AuNPs using Ganoderma spp. extract as a reducing and stabilizing agent. The as-prepared AuNPs were characterized via UV-vis, XRD, FTIR, EDX, DLS, and TEM. The biologically derived AuNPs were spherical, discrete, and the average size was 20 nm. The biocompatibility effect of AuNPs was investigated using cell viability, LDH, and ROS assays. The results indicate that biologically derived AuNPs are biocompatible. Finally, this eco-friendly method provides an alternative route for large-scale production of biocompatible AuNPs that can be used in catalysis, sensors, electronics, and biomedical applications, especially for cancer therapy. Acknowledgements This work was supported by the KU-Research Professor Program of Konkuk University. Dr Sangiliyandi Gurunathan was supported by a Konkuk University KU-Full-time Professorship.

Similar results were observed with the in vivo experiments as well. Although fewer pups died within 24 hrs post-infection in the GANT61 chemical structure groups infected with RS218cured as compared to the groups infected with wtRS218 and RS218compl, there was no statistically significant difference in mortality rates between the three groups (Figure 5B). No mortalities were detected in the negative control group treated with

PBS or E. coli DH5α. In groups infected with wtRS218 or RS218compl, 84-87% of rat this website pups that survived 24 hrs post-infection showed septicemia, whereas in groups treated with RS218cured strain, only 33% had septicemia. In all three groups the number of bacteria in the blood was too numerous to count (>1.5-2.8 *104 CFU/ml). Also, E. coli were re-isolated Ilomastat from CSF collected from 84-87% of rat pups infected with wtRS218 or RS218compl whereas only 29% CSF samples collected from rat pups infected with RS218cured strain contained E. coli suggesting a role of pRS218 in translocation of bacteria through the blood brain barrier (BBB) to cause meningitis. Similarly, histopathological

evaluation of brain tissue from the rat pups inoculated with wRS218 or RS218compl strains demonstrated lesions consistent with meningitis (Figure 6). The bacterial loads in CSF were 4.57 + 3.02*103 in rat pups infected with wtRS218 strain and 3.77 + 2.24*103 in rat pups infected with RS218cured strain. Figure 4 Confirmation of pRS218 curing. A, Plasmid profiles of wtRS218 and RS218cured. B, PCR amplification of selected pRS218 genes in wtRS218 and RS218cured. Lane 1,100 bp ladder; Lane 2, senB; Lane 3, scsD; Lane 4, transposase; Lane 5, traU; Lane 6, pRS218_113; Lane 7, ycfA; Lane 8, ompA. C, Adenosine triphosphate Growth of wtRS218 and RS218cured E. coli in LB broth, M9 medium containing 10 μg/ml niacin broth (M9),

and complete cell culture medium (CM). Figure 5 Evaluation of virulence potential of pRS218 in vitro and in vivo. A, Involvement of pRS218 in invasion of hCMEC cells. B, Comparison of mortality, septicemia and meningitis among the groups of rat pups infected with wtRS218, RS218cured, RS218compl. ** denotes statistical significance and * denotes no statistical significance. Figure 6 Histopathological evaluation of brain tissue from rat pups. Five-day-old rat pups were infected by the IP route with wtRS218, RS218compl, RS218cured, E. coli DH5α or PBS. Pups that survived were euthanized 24 hrs post-infection, and the brains were excised, embedded in formalin, sectioned in paraffin, and stained with haematoxylin and eosin. A-F: meningitic lesions observed in pups infected with wtRS218 (A and B) or RS218compl (C, D, E, and F). Arrows indicate rod-shaped bacteria in meninges and brain tissue (black), neutrophilic infiltration/neutrophilia (blue), and cerebral edema (orange). G to I: normal histology of brain tissue from pups inoculated with RS218cured (G), PBS (H) or DH5α (I).

16 [22, 24]. f c of the CCTO/Au system was larger than the calculated value (0.16). However, the critical exponent (q ≈ 0.55) was lower than the lower limit of the normal range (q ≈ 0.8 to 1), indicating a slow increase in ϵ′ with increasing metal content.

Deviation of f c and q from percolation theory may be due to the agglomeration of Au NPs to form large MK0683 Au particles in the CCTO matrix, as clearly seen in Figure 2d. f c of the CCTO/Au system is comparable to those observed in the Ba0.75Sr0.25TiO3/Ag (f c = 0.285) [9] and BaTiO3/Ni (f c = 0.232 to 0.310) [4, 7] microcomposite systems. In the cases of the nanocomposite systems of PbTiO3/Ag [8] and Pb0.4Sr0.6TiO3/Ag [11], f c values were found to be 0.16. Actually, the obtained f c and q might not be highly accurate values or not the best values due to a large range of Au NPs volume fraction between 0.1 and 0.2. However, one of the most important factors for the observed higher f c https://www.selleckchem.com/products/DAPT-GSI-IX.html for the CCTO/Au system clearly suggested a morphology transition from nanocomposite to microcomposite as Au NP concentration was increased to 20 vol.%. This result is consistent to the microcomposite systems of Ba0.75Sr0.25TiO3/Ag [9] and BaTiO3/Ni [4, 7]. Generally, the distribution of SN-38 datasheet fillers in a matrix has

an influence on the value of f c. For spherical fillers, f c of randomly distributed 3-oxoacyl-(acyl-carrier-protein) reductase fillers is given by the ratio between the particle size of the matrix phase (R 1) and the filler (R 2) [22]. When R 1/R 2 ≈ 1 or R 1 ≈ R 2, we obtain f c  ≈ 0.16. As R 1/R 2 > > 1 or R 1 > > R 2, the fillers fill the interstitial space between the matrix phase particles, resulting in a continuous percolating cluster of the filler at f c  < 0.16.

As shown in Figure 2, the particle size of CCTO (R 1) is larger than that of Au NPs (R 2), i.e., R 1/R 2 > > 1. Theoretically, f c of the CCTO/Au NP system should be lower than 0.16. However, the observed f c value in the CCTO/Au system was found to be 0.21. Therefore, it is strongly indicated that the primary factor that has a great effect on f c is the agglomeration of the Au filler. Figure 3 The dependence of Au volume fraction on ϵ′ at RT for CCTO/Au nanocomposites. The symbols and solid curve represent the experimental data and the fitted curve, respectively. Insets 1 and 2 show the frequency dependence of ϵ′ at RT and tanδ (at 1 kHz and RT) of CCTO/Au nanocomposites. Large increases in ϵ′ of percolating composites are generally attributed to formation of microcapacitor networks in the composites and/or Maxwell-Wagner polarization [4, 9, 22]. For pure CCTO ceramics, the giant dielectric response is normally associated with the mean grain size [16, 17, 25].

Actors are not entirely free, but embedded (Garud and Karnøe 2003; Garud et al. 2007). Entrepreneurs may need to ‘run in packs’, which means coordinating their actions find more to simultaneously pursue their own and collective interests, and simultaneously cooperating and competing with others as they develop and commercialize their new ventures (Van de Ven 2005). As the numbers of entrepreneurs grow, a complex network of cooperative and competitive

relationships begins to generate critical mass and produce effective collective action. This infrastructure includes institutional arrangements to legitimate, regulate, and standardize a new technology; public resource endowments of basic scientific knowledge, financing mechanisms, and a pool of competent labor; the creation and development of markets, consumer education and demand, proprietary MK-8776 mw R&D, and the development of manufacturing, production, and distribution functions by private entrepreneurial firms

to commercialize an innovation for profit. This infrastructure may be developed by superstructure organizations often specializing in coordinating flows of information or coordinating the activities of substructure organizations (Van de Ven 1993, 2005; Jacobsson and Johnson 2000). Concerted action from Selleckchem S3I-201 different social enterprises and the mobilization of support from multiple other actors in the innovation system for the diffusion Bay 11-7085 and legitimization of new institutional arrangements might, thus, be key requirements for social enterprises that aim to upscale their businesses for solar home systems in India. This is also recognized in a

related stream of literature that aims to understand how advocates of radical, potentially more sustainable technologies gain increasing support for their technologies. This literature under the heading of strategic niche management (SNM) is part of evolutionary approaches to understanding systemic transformation in socio-technical systems towards sustainability (Kemp et al. 1998). In SNM, innovations with promising sustainability characteristics are conceptualized as emerging and developing in ‘niches’, i.e., emerging institutional environments that provide a (partially) protected space in which actors experiment and incubate promising concepts or prototypes. The relation between the emerging institutional environment, the space it generates, and the activities performed by innovating actors within that space is conceptualized as cyclic and co-evolutionary. Experiments represent small initiatives in which the earliest stages of socio-technical learning and co-evolution take place. Experiments typically bring together new networks of actors with knowledge, capabilities, and resources, who cooperate in a process of social learning (Berkhout et al. 2010).

Further, we investigated the antitumor activity of AZD8931 alone or in combination with paclitaxel in EGFR-overexpressed and HER2 non-amplified IBC models. Methods and materials Reagents and cell culture AZD8931 was synthesized and generously provided

by AstraZeneca [16]. SUM149 were obtained from Dr. Stephen Ethier (Kramanos Institute, MI, USA) and #CB-839 solubility dmso randurls[1|1|,|CHEM1|]# are commercially available (Asterand, Detroit, MI). SUM149 cells were cultured in Ham’s F-12 media supplemented with 10% fetal bovine serum (FBS), 1 μg/ml hydrocortisone, 5 μg/ml insulin and antibiotic-antimycotic. The FC-IBC-02 tumor cells were derived from primary human breast cancer cells isolated from pleural effusion of an IBC patient [14, 15]. Human samples used in this study were acquired with approval of the Fox Chase PF-562271 chemical structure Cancer Center’s Institutional Review Board. Importantly, written

informed consent was obtained from each participant. FC-IBC-02 cells were cultured in DMEM/F12 media with 10% FBS and 1% L-glutamine and antibiotic-antimycotic. Antibodies and immunoblot Following treatment with AZD8931 at the indicated concentration and time points, immunoblotting was performed as previously described [15]. In brief, cells were lysed in 1× lysis buffer (Cell signaling), and then the supernatant was collected by centrifuging at 10,000 rpm for 10 min at 4°C. Protein concentration was determined using the BCA protein assay reagent kit (Pierce, Rockford, IL). Equal amounts of protein from cell lysates were resolved by SDS-PAGE electrophoresis. The membranes were incubated at 4°C overnight with the following antibodies: mouse anti-EGFR (1:1000; Cell Signaling), rabbit anti-AKT and rabbit anti-phospho-AKT (1:1000; Cell Signaling), mouse anti-β-actin (1:5,000; Santa Cruz). After incubation with anti-mouse IgG horseradish peroxidase conjugated secondary antibody (1:5,000; Amersham Pharmacia Biotech), immunoreactive proteins were visualized by the enhanced chemiluminescence reagents. Cell proliferation and apoptotic assay SUM149 and FC-IBC-02

cells (2 × 103) were seeded in triplicate in a 96-well plate and cultured overnight. Cells were treated TCL with AZD8931 at indicated concentration for 72 hrs. Cell proliferation was monitored at the indicated times, absorbance at 490 nm was measured using a microplate reader using the MTS assay (CellTiter 96 AQueous One Solution cell proliferation assay, Promega) according to the manufacturer’s instruction. Apoptotic cells were measured by Annexin V staining. Cells (1 × 105) were treated with 1 μM AZD8931 for 48 and 72 hrs. Cells were harvested and labeled with Annexin V-PE and 7-amino-actinomycin D (7-AAD) (Guava Technologies Inc, Burlingame, CA) according to the manufacturer’s instructions. The samples were then analyzed by Guava system on a GuavaPC personal flow cytometer (Guava Technologies).

The sample Cy5-dye labelled cDNAs and the reference Cy3-dye labeled cDNAs were mixed (1:1) and purified for removal of uncoupled dye by using a QIAquick PCR purification kit (Qiagen, Valencia, CA), as described by the supplier. The pellets obtained were dissolved in 35 μl hybridization buffer (5x SSC, 0.2% SDS, 5x Denhardt’s solution, 50% (v/v) formamide and 0.2 ug/ul denatured herring-sperm DNA), boiled for 5 min and spun down briefly. Networks construction and analysis A bipartite

network, named Network 1 was constructed Selleckchem AZD6738 with the novo generated gene expression data in this study by connecting two sets of nodes: one set was formed by genes differentially transcribed under several culture conditions. The other set of nodes included the environmental conditions (heat, find more oxidative and acid stress in anoxic and oxic condition, osmotic stress under anoxic condition and non-stressing anoxic conditions) Selleck Anlotinib combined with the regulation pattern, i.e. up or down-regulation. Network 2 was constructed by extending network with nodes representing genes and conditions to include the transcriptional response reported during the lag period,

exponential growth and stationary phase [7] and in immobilized cultures in different stages [8, 9]. Network 3 was a bipartite genome scale network including all genes in the genome of S. Typhimurium LT2 and plasmids of S. Typhimurium SL1344 as previously described [10]. Edges connected two sets of nodes. Genes constituted one of these sets of nodes. The genome composition was obtained from the Genome Project NCBI database [65]. The other set of nodes included metabolic pathways and cellular functions, according to the KEGG database [66], the CMR-TIGR database [67] and the COGs (Clusters of Orthologous Groups of proteins) functional categories obtained from the Genome Project NCBI database [65]. The number of nodes was 5153, from

which 4717 were genes and the remaining 436 nodes represented metabolic pathways and cellular functions. There were 11626 edges between these two sets of nodes. For networks representation and topological quantification we used the programs PAJEK [68] and Cytoscape [69]. Networks modularity was estimated implementing CYTH4 the fast modularity maximization algorithm [11]. Cluster analysis Hierarchical clustering was performed using the SAS 9.2 software [70] on the novo generated microarray data in this work using the Unweighted Pair Group Method with Arithmetic Mean (UPGMA). Expression values were coded as 1 if genes were induced, -1 if repressed and 0 if not affected. Environmental conditions (heat, oxidative and acid stress in anoxic and oxic condition, osmotic stress under anoxic condition and non-stressing anoxic conditions) were clustered according to the gene expression values. Construction of mutants Cultures were grown in LB broth (Oxoid, CM1018) or on solid media consisting of LB-broth with addition of 1.

In the present study, we also showed that after 28 days of heavy selleckchem resistance training and supplementation NO underwent increases in myofibrillar protein of 70.39% that were significantly greater than the 26.34% increase in PL (p < 0.001), and that the increases for NO were significantly different than PL (p = 0.014). This is a similar pattern of response from longer-term studies where creatine supplementation, in conjunction with 12 wk of resistance training, resulted in a 57.92% increase in myofibrillar protein content when

compared to a maltodextrose placebo group, which only increased 11.62% [24]. In addition, 10 wk of heavy resistance training combined with a protein and amino acid supplement resulted in a 25.03% increase in myofibrillar protein compared to 10.54% for a carbohydrate placebo [34]. We have demonstrated 28 days of heavy resistance training to increase serum IGF-1 by 9.34% www.selleckchem.com/products/elacridar-gf120918.html and 8.58%, respectively for NO and PL; however, GSK2118436 cost there

was no difference between groups. Treating C2C12 myoblasts with creatine has been shown to increase the expression of the IGF-1 peptide [40]. A positive relationship has been reported between IGF-1 peptide and total DNA content in muscle during resistance exercise due to satellite cell proliferation stimulated by the locally produced IGF-1 [7]. However, while the IGF-I peptide expressed in skeletal muscleincreases muscular protein synthesis and stimulates differentiation of proliferating satellite cells [14, 41], it is unclear whether increases in hepatically-derived circulating IGF-1 has any direct effect on muscle hypertrophy. We have previously shown that 10 wk of heavy resistance training combined with a daily supplement containing whey/casein protein and free amino acids increased circulating IGF-1 levels, while also increasing muscle strength and mass [34]. Additionally, 16 wk of resistance training has been shown to increase circulating IGF-1 levels [42]. However, 12 wk of heavy resistance training has been shown to increase muscle strength and mass without any corresponding

increases in circulating IGF-1 [43]. Increases in muscle hypertrophy independent of increases in circulating IGF-1 can possibly be explained by a recent study using a liver IGF-1 deficient mouse model, which Chloroambucil involves a reduction in serum IGF-1 of approximately 80% [44]. After 16 wk of resistance training, the IGF-1-deficient mice and control mice exhibited equivalent gains in muscle strength, suggesting that performance and recovery in response to resistance training is normal even when there is a severe deficiency in circulating IGF-1. HGF is a growth factor bound to an extracellular matrix in skeletal muscle [45] that is capable of activating quiescent satellite cells [46]. Serum HGF levels have been shown to increase 24 hr following a single bout of eccentric exercise [47].

Variable Time Point WP CHO p-value IRS-1 Baseline 15.68 ± 9.6 19.52 ± 6.4 Supplement (S) = 0.88   15 min CB-839 in vitro post-exercise 29.04 ± 6.6† 22.28 ± 11.2 Test (T) = 0.04†#   120 min post-exercise 25.40 ± 6.0 19.65 ± 9.2 S × T = 0.44 Akt Baseline 5.04 ± 1.9 6.88 ± 1.1 Supplement (S) = 0.21   15 min post-exercise 6.04 ± 2.6 5.61 ± 4.1 Test (T) = 0.35   120 min post-exercise

4.78 ± 1.4 4.58 AR-13324 chemical structure ± 2.1 S × T = 0.82 mTOR Baseline 3.34 ± 0.34 3.62 ± 0.19 Supplement (S) = 0.93   15 min post-exercise 3.75 ± 0.62 3.66 ± 0.27 Test (T) = 0.002†   120 min post-exercise 3.33 ± 0.19 3.52 ± 0.28 S × T = 0.34 P70S6K Baseline 8.51 ± 3.2 10.41 ± 3.2 Supplement (S) = 0.96   15 min post-exercise 14.14 ± 6.6 11.18 ± 2.9 Test (T) = 0.04   120 min post-exercise 13.32 ± 6.1 11.24 ± 5.0 S × T = 0.74 4E-BP1 Baseline 4.30 ± 2.4 5.33 ± 1.7 Supplement (S) = 0.28   15 min post-exercise 2.66 ± 1.3† 2.28 ± 1.0 Test (T) = 0.001†   120 min post-exercise 4.07 ± 1.9# 4.90 ± 1.8 S × T = 0.64 Data are means ± standard deviations. mTOR is expressed as absorbance units at 450 nm/mg. † represents significant difference from baseline at 15 JIB04 in vitro min post-exercise.

# represents significant difference from baseline at 120 min post-exercise. Discussion In the present study, we chose to assess changes in the activity of Akt/mTOR pathway intermediates as markers of MPS in response to resistance exercise after ingesting 10 g of whey protein. As a result, we observed resistance exercise to effectively activate signaling PIK3C2G intermediates of the Akt/mTOR pathway. Specifically, we demonstrated increased phosphorylation of IRS-1, AKT, and mTOR. Relative to their downstream targets, p70S6K was hyper-phosphorylated at 15 min post-exercise, whereas 4E-BP1 was hypo-phosphorylated at 15 min post-exercise. Conversely, we also observed that ingesting 10 g of whey protein was unable to induce a greater response in such kinase phosphorylation when compared to ingesting carbohydrate. Therefore, our results

suggest that ingestion of 10 g of whey protein (5.25 g EAAs) is no different than an equal amount of carbohydrate at enhancing the activity of systemic and cellular signaling markers indicative of MPS following resistance exercise. Resistance exercise and amino acids effectively stimulate MPS [30]. Based on previous studies, the role that nutrient ingestion plays in activating the Akt/mTOR pathway [15, 18–20] is not completely understood, and may likely be related to the amount of amino acids available or whether co-ingested with carbohydrate. Previous studies have demonstrated that 20 g of whey protein (8.6 g EAAs) [10] and 10 g EAAs [26] maximally stimulated MPS, but that MPS was also increased even at whey protein doses of 5 g (2.2 g EAAs) and 10 g (4.3 g EAAs) [10] and an EAA dose of 5 g [26].