Effects of Cycling Intensity on Acute Signaling Adaptations to 8-weeks Concurrent Training in Trained Cyclists.

This study examined whether the intensity of endurance stimuli modifies the adaptation in strength and endurance following concurrent training and whether the acute molecular response to concurrent exercise is affected by training status. Using a parallel group design, trained cyclists were randomized to either resistance exercise followed by moderate intensity continuous training (RES + MICT, n = 6), or resistance exercise followed by work matched high intensity interval training (RES + HIIT, n = 7), across an 8 weeks training programme. A single RES + MICT or RES + HIIT exercise stimulus was completed 1 week before and within 5 days of completing the training programme, to assess phosphorylation of protein kinases of the mTOR and AMPK signaling pathways. There were no main effects of time or group on the phosphorylation of protein kinases in response to concurrent exercise stimulus pre- and post-training intervention (p > 0.05). Main effects of time were observed for all maximal strength exercises; back-squat, split-squat, and calf-raise (p < 0.001), with all improving post intervention. A time × group interaction was present for V̇O2peak, with the RES + MICT group displaying a preferential response to that of the RES + HIIT group (p = 0.010). No time nor group effects were observed for 5 min time trial performance, power at 2 and 4 mmol L-1 (p > 0.05). Whilst preliminary data due to limited sample size the intensity of endurance activity had no effect on performance outcomes, following concurrent training. Further, the acute molecular response to a concurrent exercise stimulus was comparable before and after the training intervention, suggesting that training status had no effect on the molecular responses assessed.

Aerobic exercise intensity does not affect the anabolic signaling following resistance exercise in endurance athletes.

This study examined whether intensity of endurance stimulus within a concurrent training paradigm influenced the phosphorylation of signaling proteins associated with the mTOR and AMPK networks. Eight male cyclists completed (1) resistance exercise (RES), 6 × 8 squats at 80% 1-RM; (2) resistance exercise and moderate intensity cycling of 40 min at 65% V̇O2peak, (RES + MIC); (3) resistance exercise and high intensity interval cycling of 40 min with 6 alternating 3 min intervals of 85 and 45% V̇O2peak (RES + HIIC), in a cross-over design. Muscle biopsies were collected at rest and 3 h post-RES. There was a main effect of condition for mTORS2448 (p = 0.043), with a greater response in the RES + MIC relative to RES condition (p = 0.033). There was a main effect of condition for AMPKα2T172 (p = 0.041), with a greater response in RES + MIC, relative to both RES + HIIC (p = 0.026) and RES (p = 0.046). There were no other condition effects for the remaining protein kinases assessed (p > 0.05). These data do not support a molecular interference effect in cyclists under controlled conditions. There was no intensity-dependent regulation of AMPK, nor differential activation of anabolism with the manipulation of endurance exercise intensity.

Characterization of a selective and potent antagonist of human P2X(7) receptors, AZ11645373.

BACKGROUND AND PURPOSE: The ATP-gated P2X(7) receptor has been shown to play a role in several inflammatory processes, making it an attractive target for anti-inflammatory drug discovery. We have recently identified a novel set of cyclic imide compounds that inhibited P2X(7) receptor-mediated dye uptake in human macrophage THP-1 cells. In this study the actions and selectivity of one of these compounds, AZ11645373, were characterized. EXPERIMENTAL APPROACH: We measured membrane currents, calcium influx, and YOPRO-1 uptake from HEK cells expressing individual P2X receptors, and YOPRO1 uptake and interleukin-1beta release from THP-1 cells in response to ATP and the ATP analogue benzoylbenzoyl ATP (BzATP). KEY RESULTS: AZ11645373 up to 10 microM, had no agonist or antagonist actions on membrane currents due to P2X receptor activation at human P2X(1), rat P2X(2), human P2X(3), rat P2X(2/3), human P2X(4), or human P2X(5) receptors expressed in HEK cells. AZ11645373 inhibited human P2X(7) receptor responses in HEK cells in a non-surmountable manner with K (B) values ranging from 5 - 20 nM, with mean values not significantly different between assays. K (B) values were not altered by removing extracellular calcium and magnesium. ATP-evoked IL-1beta release from lipopolysaccharide-activated THP-1 cells was inhibited by AZ11645373, IC(50) = 90 nM. AZ11645373 was > 500-fold less effective at inhibiting rat P2X(7) receptor-mediated currents with less than 50% inhibition occurring at 10 microM. CONCLUSIONS AND IMPLICATIONS: AZ11645373 is a highly selective and potent antagonist at human but not rat P2X(7) receptors and will have much practical value in studies of human cells.

Novel P2X7 receptor antagonists.

The synthesis and pharmacological evaluation of a new series of potent P2X(7) receptor antagonists is disclosed. The compounds inhibit BzATP-mediated pore formation in THP-1 cells. The distribution of the P2X(7) receptor in inflammatory cells, most notably the macrophage, mast cell and lymphocyte, suggests that P2X(7) antagonists have a significant role to play in the treatment of inflammatory disease.

Identification of Antheridiogens in Lygodium circinnatum and Lygodium flexuosum.

Antheridiogens in two species of Schizaeaceous ferns, Lygodium circinnatum and Lygodium flexuosum, were analyzed by gas chromatography-mass spectrometry. In L. circinnatum, gibberellin A73 (GA73) methyl ester (GA73-Me), which had originally been identified in L. japonicum, was identified as a principal antheridiogen, and the methyl esters of five known GAs (GA9, GA20, GA70, GA88, and 3-epi-GA88) were also identified as minor antheridiogens. In addition, four compounds corresponding to isomers of monohydroxy-GA73-Me were detected. One of these was shown to be 12[beta]-hydroxy-GA73-Me, the parent acid of which has been allocated the GA assignment GA96. The other three compounds, tentatively named X1, X2, and X3, have not been fully characterized. In L. flexuosum, GA73-Me was also identified as a major antheridiogen, with X2 being detected as a minor one. The total antheridium-formation activity in the culture medium of 7-week-old prothallia of L. circinnatum and L. flexuosum was more than 1000 times higher than that of L. japonicum. On the other hand, the response of gametophytes of the former two Lygodium ferns to GA73-Me was more than 100 times lower than that of L. japonicum.