The seeds of three Eucalyptus species (Myrtaceae) (Origin: Australia), including:, Eucalyptus loxophleba Benth., Eucalyptus microtheca F.Muell. and Eucalyptus largiflorens F.Muell. were cultivated in 1993-1994 at two research stations (Shushtar and Dezful) in Khuzistan province (western south region of Iran) and a research station (Kashan) in Isfahan province (Central region of Iran). These species have good adaptability to climatic condition of this area (dry warm). The leaves of these species were collected in middle of the four seasons (spring, summer, autumn and winter) at two consecutive years, in order to find out the best harvesting time with the highest oil yield and 1,8-CINEOLE percentage. After drying the plant materials in shade, their essential oils were obtained by hydro-distillation.The oils were analyzed by GC and GC/MS devises. The results showed that the harvesting time had significant effects on oil yield and 1,8-CINEOLE content of the Eucalyptus species. The main components of the harvested oils consisted of 1, 8-CINEOLE and a-pinene. The highest oil yields of E. loxophleba Benth. in all localities were obtained from the samples collected in autumn and winter. The highest percentage of 1,8-CINEOLE for Dezful site was extracted from the winter (70.2%) and autumn (63.2%) samples, For Kashan sits, was extracted from the winter (55.5%) and spring (58.1%) samples and for Shushtar site, was extracted from the summer samples (53.6 and 49.6%). The highest oil yield of E. microtheca F.Muell. was obtained in spring from Dezful and Shushtar localities and in autumn from Kashan locality. For both Shushtar and Dezful sites, the highest percentage of 1,8-CINEOLE was obtained in winter and summer samples whereas for Kashan site, was obtained in summer samples. The highest oil yield of E. largiflorens F.Muell. was obtained in autumn and winter for Dezful and Shushtar samples and summer for Kashan samples. The highest percentage of 1,8-CINEOLE was extracted in summer and winter for Shushtar samples, in spring and winter for Dezful samples and winter for Kashan samples.

The Alzheimer’, s disease is one of the neurodegenerative diseases affecting many individuals around the world with no definitive cure. The aim of this study was to investigate the effect of CINEOLE, as a natural antioxidant, on the behavioral, biochemical, and histological symptoms of Alzheimer’, s disease in Wistar rats. Thirty-six male Wistar rats were randomly assigned to six groups. The control group underwent any surgery with no special treatment. The other groups underwent surgery and were assigned to four beta-amyloid groups: one group receiving PBS (beta-amyloid solvent),the sham group was treated with tween 80 (CINEOLE solvent) and the two experimental groups were treated with CINEOLE at 0. 02 and 0. 04 mg/kg doses. The control group, the Alzheimer’, s group and the PBS group did not receive any treatments. Finally, behavioral (Shuttle box), biochemical (malondialdehyde (MDA), superoxide dismutase, TNF-α, ) and histological parameters (H&E and Thioflavine S staining) were investigated. In Alzheimer’, s-induced rats, the time it took for the rats to go to the dark part of the shuttle box was significantly (p < 0. 001) shorter than that of the control group. The levels of malondialdehyde and TNF-α,in these rats were significantly increased and the level of superoxide dismutase was significantly decreased compared to the control group (p < 0. 001). Furthermore, histological studies showed that in Alzheimer’, s-induced rats, neuronal cell death occurred extensively in the hippocampus and amyloid plaques increased. Treatment of rats with CINEOLE improved all of the investigated parameters significantly in a dose-dependent manner. CINEOLE may be used as a suitable natural antioxidant to treat Alzheimer’, s disease and reduce its symptoms, requiring further research.

The aim of the present study was to investigate the influence and the mechanisms of CINEOLE and terpineol on the in-vitro transdermal delivery of huperzine A from microemulsions, and their potential synergistic effect on the permeation enhancement. The transdermal delivery of huperzine A from microemulsions with different concentrations of CINEOLE and terpineol through the rat abdominal skin was determined by Franz-type diffusion cells. The partition coefficient of huperzine A between the full thickness skin and microemulsion was determined. Attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR) was carried out to analyze the effects of CINEOLE and terpineol on the biophysical properties of the stratum corneum (SC) and the mechanisms of permeation enhancement. These results indicated that CINEOLE and terpineol could synergistically increase the transdermal delivery of huperzine A from microemulsions through increasing the partition and diffusion coefficients of huperzine A. ATR-FTIR studies further validated the synergistic effect and revealed that the enhancing mechanisms were due to increasing the disorderliness and fluidity of SC lipid alkyl chains, disrupting the structure of keratin in SC, and extracting SC lipids. In conclusion, CINEOLE and terpineol, acting synergistically to enhance the transdermal delivery of huperzine A from microemulsions, might provide an alternative permeation enhancer combination for the transdermal delivery of huperzine A.

Introduction: To the best of our knowledge, there is little information in regards to the cytotoxicity of 1, 8-CINEOLE. Insect cells exhibit wide resistance to the lethal effects of chemical compounds. Activation of p53 by acetylation can cause cell toxicity. This study has been carried out in order to investigate whether the use of 1, 8-CINEOLE in the Spodoptera frugiperda SF-9 cell line can induce cytotoxicity by increasing p53 acetylation expression or not. Materials and Methods: SF-9 cell line was cultured in TC100 insect culture medium and treated with 1, 8-CINEOLE at concentration of 850. 45 μ mol/L, based on the half-maximal inhibitory concentration (IC50 ) index at different times (24, 48 and 72h). The IC50 value was estimated for 1, 8-CINEOLE in SF-9. The percentage of alive cells was measured by MTT assay. The ELISA and Bradford protein techniques were used to detect endogenous levels of total and acetylated p53 protein generated in SF-9 cells. Results: The findings of the present study indicated that treatment with 850. 45 μ mol per liter of 1, 8-CINEOLE shows a time-dependent increase in the number of dead cells of SF-9 cell line. The effect of severe toxicity on SF-9 was observed after 72 h of incubation with 1, 8-CINEOLE, with approximately 4% of SF-9 cells alive. We observed a significant increase in the level of p53 acetylation up to 48 h in SF-9, indicating that 1, 8-CINEOLE resulted in up-regulation of acetylated P53 and consequently p53 activation in SF-9 cells. Results showed that there is relationship between acetylated p53 protein levels and 1, 8-CINEOLE toxicity in SF-9 cell line. Conclusions: 1, 8-CINEOLE, may function through common pathways and mediate their cytotoxic effects through targeting p53 and its acetylation in SF-9 cells.