Dengue virus (DENV) infection and replication induces oxidative stress, which further contributes to the progression and pathogenesis of the DENV infection. Modulation of host antioxidant molecules may be a useful strategy for interfering with DENV replication. In this study, we showed that induction or exogenous overexpression of heme oxygenase-1 (HO-1), an antioxidant enzyme, effectively inhibited DENV replication in DENV-infected Huh-7 cells. This antiviral effect of HO-1 was attenuated by its inhibitor tin protoporphyrin (SnPP), suggesting that HO-1 was an important cellular factor against DENV replication. Biliverdin but not carbon monoxide and ferrous ions, which are products of the HO-1 on heme, mediated the HO-1-induced anti-DENV effect by non-competitively inhibiting DENV protease, with an inhibition constant (Ki) of 8.55 ± 0.38 μM. Moreover, HO-1 induction or its exogenous overexpression, rescued DENV-suppressed antiviral interferon response. Moreover, we showed that HO-1 induction by cobalt protoporphyrin (CoPP) and andrographolide, a natural product, as evidenced by a significant delay in the onset of disease and mortality, and virus load in the infected mice’s brains. These findings clearly revealed that a drug or therapy that induced the HO-1 signal pathway was a promising strategy for treating DENV infection.

SummaryThe arginine-specific protease activity of Porphyromonas gingivalis is considered to be an important factor in the pathogenic potential of this organism in destructive periodontal disease. Multiple forms of closely related Arg-x proteases are present in the culture supernatants of P. gingivalis W50. RI is a heterodimer (␣/␤) in which the catalytic ␣ chain is associated with a second ␤ chain which functions as a haemagglutinin. RIA is a single-chain enzyme (␣) and RIB is a highly post-translationally lipid-modified enzyme (LPS-␣) with reduced solubility compared to the other two forms. The N-terminal sequence of the ␣ chain of all three forms is identical, suggesting that all these enzymes may arise by differential processing of the prpR1 (protease polyprotein for RI ). In the present study we constructed a prpR1 ¹ strain of P. gingivalis W50 by insertional gene inactivation and characterized the residual extracellular Arg-x protease activity of the resulting mutant. Loss of prpR1 expression led to the abolition of RI, RIA and RIB but the total Arg-x activity in the supernatant of this strain was reduced by only c. 66%. The remaining activity was composed of two novel forms of Arg-x protease (RIIA and RIIB) which appeared to be structurally and kinetically almost identical to RIA and RIB, respectively, except for two amino acid differences in the N-terminus at position 8 (Q→E) and position 17 (A→P) and with respect to their stability to high pH. Confirmation that RIIA and RIIB are the products of a homologous locus (prR2 ) was obtained by cloning and sequencing the prR2 which showed the predicted substitutions in the deduced translation. These data indicate that RI, RIA and RIB are produced by prpR1 expression and a maturation pathway which can give rise to a dimer and an unmodifed-or LPS-modified catalytic monomer. Furthermore, RIIA and RIIB, the products of prR2, are exported into the culture supernatant in the absence of prpR1 expression and these forms may also contribute to the pathogenic potential of this organism in destructive disease.

BACKGROUND AND PURPOSEThis study aimed to evaluate the anti-hepatitis C virus (HCV) activity of andrographolide, a diterpenoid lactone extracted from Andrographis paniculata, and to identify the signalling pathway involved in its antiviral action. EXPERIMENTAL APPROACHUsing HCV replicon and HCVcc infectious systems, we identified anti-HCV activity of andrographolide by measuring protein and RNA levels. A reporter activity assay was used to determine transcriptional regulation of anti-HCV agents. A specific inhibitor and short hairpin RNAs were used to investigate the mechanism responsible for the effect of andrographolide on HCV replication. KEY RESULTSIn HCV replicon and HCVcc infectious systems, andrographolide time-and dose-dependently suppressed HCV replication. When combined with IFN-α, an inhibitor targeting HCV NS3/4A protease (telaprevir), or NS5B polymerase (PSI-7977), andrographolide exhibited a significant synergistic effect. Andrographolide up-regulated the expression of haeme oxygenase-1 (HO-1), leading to increased amounts of its metabolite biliverdin, which was found to suppress HCV replication by promoting the antiviral IFN responses and inhibiting NS3/4A protease activity. Significantly, these antiviral effects were attenuated by an HO-1-specific inhibitor or HO-1 gene knockdown, indicating that HO-1 contributed to the anti-HCV activity of andrographolide. Andrographolide activated p38 MAPK phosphorylation, which stimulated nuclear factor erythroid 2-related factor 2 (Nrf2)-mediated HO-1 expression, and this was found to be associated with its anti-HCV activity. CONCLUSIONS AND IMPLICATIONSOur results demonstrate that andrographolide has the potential to control HCV replication and suggest that targeting the Nrf2-HO-1 signalling pathway might be a promising strategy for drug development.

No abstract

BACKGROUND: There are alterations in strength in relation to menstrual cycle phase but little data attributing these responses to female sex hormone levels using a pseudo-menstrual cycle group as control. OBJECTIVE: Examining the effects of menstrual cycle phase on the development of peak torque across a range of isokinetic speeds. METHOD: 17 well trained females, 11 formed the non-oral contraceptive group (n-OC) (age 20.7 ± 1.4 yrs, mass 59.2 ± 6.9 kg, height 166.8 ± 7.1 cm) and 6 the oral contraceptive control group (OC) (age 20.3 ± 0.5 yrs, mass 60.5 ± 4.2 kg, height 164.8 ± 4.8 cm). Concentric strength of the knee flexors and extensors (60-240 • /s) was assessed, corresponding to menstruation (MEN), mid-follicular (mFOL), mid-luteal (mLUT) and pre-menstrual (pMEN). RESULTS: For n-OC significant decreases in peak torque production of the extensors at 120 • /s (P = 0.0207) (MEN) and of the flexors at 60 • /s (P = 0.0116) (MEN) and 120 • /s (P = 0.0282) (MEN) were observed compared to pMEN. No significant differences were observed across any menstrual cycle phase and peak torque for the OC group (p > 0.05). Significant positive correlations were observed (mLUT) between peak torque and oestrogen at 60 • /s (P = 0.040) and 120 • /s (P = 0.031). CONCLUSIONS: There are significant fluctuations in peak torque of the knee extensors in response to phases of the menstrual cycle associated with variances in the female sex hormones. The findings have implications for the planning of strength training in female athletes.