Strawberry and Achenes Hydroalcoholic Extracts and Their Digested Fractions Efficiently Counteract the AAPH-Induced Oxidative Damage in HepG2 Cells.

Strawberry fruits are highly appreciated by consumers worldwide due to their bright red color, typical aroma, and juicy texture. While the biological activity of the complete fruit has been widely studied, the potential beneficial effects of the achenes (commonly named seeds) remain unknown. In addition, when raw fruit and achenes are consumed, the digestion process could alter the release and absorption of their phytochemical compounds, compromising their bioactivity. In the present work, we evaluated the protective effects against oxidative damage of nondigested and digested extracts from strawberry fruit and achenes in human hepatocellular carcinoma (HepG2) cells. For that purpose, cells were treated with different concentration of the extracts prior to incubation with the stressor agent, AAPH (2,2'-azobis(2-amidinopropane) dihydrochloride). Subsequently, intracellular accumulation of reactive oxygen species (ROS) and the percentage of live, dead, and apoptotic cells were determined. Our results demonstrated that all the evaluated fractions were able to counteract the AAPH-induced damage, suggesting that the achenes also present biological activity. The positive effects of both the raw fruit and achenes were maintained after the in vitro digestion process.

In vitro and in vivo antifungal activities of T-2307, a novel arylamidine, against Cryptococcus gattii: an emerging fungal pathogen.

Objectives: T-2307, a novel arylamidine, exhibits potent broad-spectrum activities against the majority of fungal pathogens. In this study, the antifungal activity of T-2307 against Cryptococcus gattii was evaluated in comparison with those of amphotericin B, fluconazole and voriconazole in vitro and in vivo . Methods: The MICs for 15 clinical isolates were determined according to CLSI guidelines and time-kill studies were performed using C. gattii YF2784. In a murine model for intranasal pulmonary infection caused by C. gattii YF2784, the test compounds were administered once daily for 7 days from 2 h or 14 days post-infection. The viable counts in the lungs and brain were determined at 21 days post-infection. Results: The MIC range, MIC 50 , MIC 90 and geometric mean MIC of T-2307 were 0.0078-0.0625, 0.0313, 0.0625 and 0.0394 mg/L, respectively. The MIC of T-2307 was significantly lower than those of fluconazole, voriconazole and amphotericin B. T-2307 showed concentration-dependent fungicidal activity at 4 times the MIC or higher. Administration of T-2307 at 2 mg/kg/day, amphotericin B at 1 mg/kg/day and fluconazole at 160 mg/kg/day from 2 h post-infection significantly reduced viable counts in the lungs and brain. However, when the administration was started 14 days post-infection, only T-2307 significantly reduced the viable counts in both the lungs and the brain at 1 mg/kg/day. Conclusions: T-2307 shows excellent in vitro and in vivo antifungal activities against C. gattii and would be a promising new candidate for the treatment of cryptococcosis.

In vitro and in vivo investigation of the efficacy of arylimidamide DB1831 and its mesylated salt form--DB1965--against Trypanosoma cruzi infection.

Chagas disease is caused by infection with the intracellular protozoan parasite Trypanosoma cruzi. At present, nifurtimox and benznidazole, both compounds developed empirically over four decades ago, represent the chemotherapeutic arsenal for treating this highly neglected disease. However, both drugs present variable efficacy depending on the geographical area and the occurrence of natural resistance, and are poorly effective against the later chronic stage. As a part of a search for new therapeutic opportunities to treat chagasic patients, pre-clinical studies were performed to characterize the activity of a novel arylimidamide (AIA--DB1831 (hydrochloride salt) and DB1965 (mesylate salt)) against T. cruzi. These AIAs displayed a high trypanocidal effect in vitro against both relevant forms in mammalian hosts, exhibiting a high selectivity index and a very high efficacy (IC(50) value/48 h of 5-40 nM) against intracellular parasites. DB1965 shows high activity in vivo in acute experimental models (mouse) of T. cruzi, showing a similar effect to benznidazole (Bz) when compared under a scheme of 10 daily consecutive doses with 12.5 mg/kg. Although no parasitological cure was observed after treating with 20 daily consecutive doses, a combined dosage of DB1965 (5 mg/kg) with Bz (50 mg/kg) resulted in parasitaemia clearance and 100% animal survival. In summary, our present data confirmed that aryimidamides represent promising new chemical entities against T. cruzi in therapeutic schemes using the AIA alone or in combination with other drugs, like benznidazole.

Protective effect of ginseng sapogenins against 2,2'-azobis (1-aminopropane) dihydrochloride (AAPH)-induced LLC-PK1 cell damage.

The effect of ginseng sapogenins, aglycone parts of ginsenosides, against oxidative damage by radical generator, 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH), in renal epithelial LLC-PK(1) cells was investigated to identify the structural characteristics of sapogenins to have renoprotective effects. Of the tested sapogenins, Δ(20(21))-protopanaxatriol showed the strongest protective effect against the AAPH-induced LLC-PK(1) cell damage. Based on the structure and stronger activity of Δ(20(21))-protopanaxatriol than the other sapogenins, the hydroxyl group in C-6 and double bond in C-20(21) position were important for renoprotective effect of sapogenin against oxidative stress.

Effect of oxygen concentration and free radicals on in vitro development of in vitro-produced bovine embryos.

The effects of free radicals and hypotaurine on the development of bovine embryos produced by in vitro fertilization of in vitro-matured oocytes were examined. Embryos that developed to the 4- to 6-cell stage after in vitro fertilization were cultured without feeder cells in TCM199 medium supplemented with 1% calf serum (CS) under either 5% CO2 in air or 5% O2, 5% CO2 and 90% N2. The percentages of blastocysts (including early, expanding, and hatched stages) that developed under 5% O2 was higher (P < .01) than the percentage of those that developed under 20% O2. The respective percentages in 5% vs 20% O2 concentration were as follows: blastocysts (d 8), 49% vs 17%; expanded blastocysts (d 8), 19% vs 6%; hatched blastocysts (d 10), 16% vs 0%. The development of embryos to blastocysts was suppressed (P < .05) when oxygen radicals were generated in culture medium by 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) under both 5% and 20% O2. The addition of hypotaurine permitted the development of embryos to blastocyst stage in medium with AAPH only under 5% O2 (P < .05), but superoxide dismutase (SOD) did not permit the development of embryos to blastocysts. Alternatively, when embryos were cultured in medium without AAPH, the rates of development into blastocysts under 20% O2 increased (P < .05) by the addition of hypotaurine and SOD. However, under 5% O2, the rates of blastocyst formation were not improved by addition of hypotaurine and SOD. Moreover, the cell numbers of blastocysts cultured in medium containing hypotaurine were greater (P < .01) than those of blastocysts cultured in medium without hypotaurine. It is concluded that hypotaurine may exert beneficial effects on in vitro development of bovine embryos under both 20% O2 and 5% O2 of gaseous conditions.