Background: Breast cancer is a major health problem that threatens the lives of millions of women worldwide each year. Most of the chemotherapeutic agents that are currently used to treat this complex disease are highly toxic with long-term side effects. Therefore, novel generation of anti-cancer drugs with higher efficiency and specificity are urgently needed. Methods: Breast cancer cell lines were treated with eugenol and cytotoxicity was measured using the WST-1 reagent, while propidium iodide/annexinV associated with flow cytometry was utilized in order to determine the induced cell death pathway. The effect of eugenol on apoptotic and pro-carcinogenic proteins, both in vitro and in tumor xenografts was assessed by immunoblotting. While RT-PCR was used to determine eugenol effect on the E2F1 and survivin mRNA levels. In addition, we tested the effect of eugenol on cell proliferation using the real-time cell electronic sensing system. Results: Eugenol at low dose (2 μM) has specific toxicity against different breast cancer cells. This killing effect was mediated mainly through inducing the internal apoptotic pathway and strong down-regulation of E2F1 and its downstream antiapoptosis target survivin, independently of the status of p53 and ERα. Eugenol inhibited also several other breast cancer related oncogenes, such as NF-κB and cyclin D1. Moreover, eugenol up-regulated the versatile cyclin-dependent kinase inhibitor p21^WAF1 protein, and inhibited the proliferation of breast cancer cells in a p53-independent manner. Importantly, these anti-proliferative and pro-apoptotic effects were also observed in vivo in xenografted human breast tumors. Conclusion: Eugenol exhibits anti-breast cancer properties both in vitro and in vivo, indicating that it could be used to consolidate the adjuvant treatment of breast cancer through targeting the E2F1/survivin pathway, especially for the less responsive triple-negative subtype of the disease.

The aim of this research paper is to test the antistaphylococcal effect of 1,8‐cineole, amoxicillin/clavulanic acid (AMC), and gentamicin, either separately or in combination against three Staphylococcus aureus strains isolated from patients suffering from osteomyelitis. This activity was tested in vitro by using the microdilution method and the checkerboard assay. The efficacy of these three antibacterial agents was then tested in vivo by using an experimental model of methicillin‐resistant S. aureus osteomyelitis in rabbits. This efficacy was assessed after four days of treatment by counting the number of bacteria in the bone marrow. The obtained results in vitro showed that the combination of the AMC with gentamicin did not induce a synergistic effect, whereas the combination of the two antibiotics with 1,8‐cineole did. This effect is stronger when AMC is combined with 1,8‐cineole as a total synergistic effect was obtained on the three strains used (FIC ≤ 0.5). In vivo, a significant reduction was noted in the number of colonies in the bone marrow when rabbits were treated with AMC associated with either 1,8‐cineole or gentamicin compared to rabbits treated with AMC, gentamicin, or 1,8‐cineole alone. These results demonstrated that 1,8‐cineole showed a synergistic effect in combination with both AMC and gentamicin, which offer possibilities for reducing antibiotic usage. Also, the AMC associated with 1,8‐cineole could be used to treat MRSA osteomyelitis.

This study aimed to evaluate the antifungal activity of oregano and clove essential oils and their main components (thymol, carvacrol and eugenol) against Penicillium digitatum, as well as against isolates of Penicillium italicum and Geotrichum candidum resistant to imazalil or guazatine, respectively. In vitro tests were conducted by a direct contact method on potato dextrose agar medium. The results indicated that the essential oils and their main components were effective in inhibiting fungal growth, with thymol displaying the highest efficacy. The antifungal activity of thymol was evaluated further on orange fruit infected experimentally with a suspension of spores of P. digitatum, G. candidum or a mixture of both fungi, using isolates resistant to chemical fungicides. Treatment of the oranges with the thymol solutions reduced or totally inhibited the development of fruit decay, depending on the thymol concentration used. These essential oils and their main components demonstrated a strong antifungal effect against Penicillium spp. and G. candidum resistant to chemical fungicides. They may have potential as active compounds for the formulation of natural preparations to use as alternatives to synthetic fungicides for the management of green, blue and sour rot in citrus fruit.

Almond skin is an important by-product in the almond processing industry, rich in potentially health-promoting phenolic compounds. The objective of this present study is to separate the skin from the almond and extract its polyphenol contents using Ultrasound-Assisted Extraction (UAE) at room temperature. Optimization was performed according to a two-variable central composite design (CCD), and the optimum combination of ultrasonic intensity and extraction temperature was obtained through multi-response optimization: ultrasonic intensity (UI), 9.47 W.cm⁻²; and temperature, 20 °C for an extraction time of 20 min. Under the above-mentioned conditions, total phenolic content was 258% higher with UAE than silent experiment. Mathematic modelling and microscopic investigations were achieved to enable understanding physical and structural effects of ultrasound on almond skins and comprehension of the mechanism behind the enhancement of mass transfer phenomena. Scanning Electron Microscopy (SEM) showed different acoustic cavitation impacts including fragmentation, sonoporation, and erosion. Extracts were analyzed by ultra-performance liquid chromatography coupled with mass spectrometry (UPLC-MS), identifying a combination of flavanols, flavanones and non-flavonoids. UAE shows no negative effect on almond proteins and lipids when compared to natural almonds (NS).

The purpose of the present study is twofold. First, it aims to evaluate the synergistic action of the ß-lactam antibiotic; AMX is associated with 1,8-cineole on six clinical isolates of ESBL-producing Escherichia coli and Klebsiella pneumoniae strains. Second, it aims to determine the effect this association has on the ESBL enzymatic resistance mechanism. The synergistic action of AMX/1,8-cineole was evaluated using partial inhibitory concentrations (PIC), determined by a microplate, a checkerboard and time–kill assays. The effect of AMX/1,8-cineole associations on the ESBL enzymatic resistance mechanism was evaluated using a new optimized enzymatic assay. This assay was based on the determination of the AMX antibacterial activity when combined with 1,8-cineole (at subinhibitory concentrations) in the presence or absence of the ß-lactamase enzyme toward a sensitive E. coli strain. The results of both checkerboard and time–kill assays showed a strong synergistic action between AMX and 1,8-cineole. The results of the enzymatic assay showed that the combination of AMX with 1,8-cineole notably influences the enzymatic resistance of the reaction by decreasing the affinity of the β-lactam antibiotic, AMX, to the β-lactamase enzyme. All obtained results suggested that the AMX/1,8-cineole association could be employed in therapy to overcome bacterial resistance to AMX while reducing the prevalence of resistance.