Effect of bioclimatic area on the composition and bioactivity of Tunisian Rosmarinus officinalis essential oils.

The chemical composition of eight Tunisian Rosmarinus officinalis L. populations (A-H) from different bioclimatic areas has been examined by gas chromatography (GC) and GC-mass spectrometry. The essential oils are characterised by high amounts of oxygenated monoterpenes (58.2-71.7%) followed by monoterpene hydrocabons (15.1-26.7%). 1,8-Cineole, camphor, α-pinene and borneol are the main representative components. The antioxidant activity was investigated by 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), ferric reducing ability power assay and ß-carotene bleaching test. Samples showed antiradical activity by inhibiting DPPH radical with IC50 values ranging from 375.3 to 592.8 µg mL(- 1) for samples F and A, respectively. Sample A also showed the most promising activity in ß-carotene bleaching test (IC50 of 31.9 µg mL(- 1)). The essential oils were also screened for acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activity. Sample G showed the highest activity against AChE (IC50 of 64.7 µg mL(- 1)) while sample D (IC50 of 29.5 µg mL(- 1)) exhibited the most potent activity against BChE.

Antioxidant activity of Tunisian Geranium robertianum L. (Geraniaceae).

The present investigation focuses on the methanolic extract obtained from Geranium robertianum L. (Geraniaceae) (Herb Robert), a herbal plant used in traditional medicine for the treatment of human and animal diseases. The antioxidant capacities of the extract were evaluated using 1,1-diphenyl-2-picrylhydrazyl radical, ß-carotene/linoleic acid and reducing power and metal chelating activity assays. The amount of total phenolic content, flavonoids and condensed tannins was very high, and the correlation between the antioxidant activity potential and total phenolic level of the extract was pointed out.

Antiproliferative activity of hexane extract from Tunisian Cistus libanotis, Cistus monspeliensis and Cistus villosus.

BACKGROUND: As a part of our investigation on Tunisian medicinal plants, we have carried out a phytochemical investigation of the hexane extracts from leaves of Cistus libanotis, C. villosus and C. monspeliensis, evualuating also their possible antiproliferative activity in vitro. RESULTS: The major compounds of hexane extracts were identified and quantified by GC-MS. The composition of the three species, although belonging to the same genus, is completely different. The antiproliferative activity was evaluated against murine monocyte/macrophages (J774.A1), human melanoma cells (A-375), and human breast cancer cells (MCF-7), showing major activity against the human melanoma cell line A-375. CONCLUSIONS: The chemical composition of the hexane extracts from the three Cistus species can be useful in the chemosystematics of this complex genus. The preliminary antiproliferative activity against human melanoma cell line A-375 deserve further investigations in order to determine the compounds, or their combinations, which are the main responsible for the antiproliferative activity and its possible mechanism(s) of action.

Studies on the antioxidant activity of the essential oil and extract of Tunisian Tetraclinis articulata (Vahl) Mast. (Cupressaceae).

This study analyses the chemical composition and in vitro antioxidant activity of both the essential oil and the 80% aqueous acetone extract of Tetraclinis articulata leaves. The GC-MS analysis of the essential oil identified 66 components that comprise 93.5% of the oil. The major constituents of the oil are: bornyl acetate (31.4%), α-pinène (24.5%) and camphor (20.3%). Antioxidant activities of the samples were determined using four different test systems, namely DPPH, ß-carotene/linoleic acid, reducing power and metal chelating activity assay. Test results from the DPPH system showed the strongest radical scavenging activity was exhibited by the 80% aqueous acetone extract (IC50 = 5.5 µg mL⁻¹), which was two times higher than the positive control (BHT). The amount of the total phenolics, flavonoids and condensed tannins was very high in the 80% aqueous acetone extracts. The correlation between the antioxidant activity potential and total phenolic level of the extract was noted.

Genetic approaches for studying transgene inheritance and genetic recombination in three successive generations of transformed tobacco.

Transgene integration into plant genomes is a complex process accompanied by molecular rearrangements. Classic methods that are normally used to study transgenic population genetics are generally inadequate for assessing such integration. Two major characteristics of transgenic populations are that a transgenic genome may harbor many copies of the transgene and that molecular rearrangements can create an unstable transgenic locus. In this work, we examined the segregation of T1, T2 and T3 transgenic tobacco progenies. Since transfer DNA (T-DNA) contains the NptII selectable marker gene that confers resistance to kanamycin, we used this characteristic in developing a method to estimate the number of functional inserts integrated into the genome. This approach was based on calculation of the theoretical segregation ratios in successive generations. Mendelian ratios of 3:1, 15:1 and 63:1 were confirmed for five transformation events whereas six transformation events yielded non-segregating progenies, a finding that raised questions about causal factors. A second approach based on a maximum likelihood method was performed to estimate recombination frequencies between linked inserts. Recombination estimates varied among transformation events and over generations. Some transgenic loci were unstable and evolved continuously to segregate independently in the T3 generation. Recombination and amplification of the transgene and filler DNA yielded additional transformed genotypes.

Antioxidant activities of the essential oils and methanol extracts from myrtle (Myrtus communis var. italica L.) leaf, stem and flower.

This study was designed to examine the chemical composition and antioxidant activity of the essential oils and methanol extracts of Myrtus communis var. italica L. leaf, stem and flower. Myrtle leaf and flower were the valuable organs for the essential oil production representing a yield of 0.61% and 0.30% (w/w), respectively. The essential oil composition of myrtle leaf and flower was characterized by high proportions of alpha-pinene, the main compound of monoterpene hydrocarbon class, with 58.05% for leaf and 17.53% for flower. Stem was rich in oxygenated monoterpenes, largely due to 1,8-cineole with 32.84%. The total phenol contents varied between different myrtle parts; leaf extract had higher total phenol content (33.67 mg GAE/g) than flower (15.70 mg GAE/g) and stem (11.11 mg GAE/g) extracts. Significant differences were also found in total tannin contents among different myrtle parts, representing 26.55 mg GAE/g in leaf, 11.95 mg GAE/g in flower, 3.33 mg GAE/g in stem. The highest contents of total flavonoids and condensed tannins were observed in stem (5.17 and 1.99 mg CE/g, respectively) and leaf (3 and 1.22 mg CE/g, respectively) extracts. The HPLC analysis indicated that the main phenolic class was hydrolysable tannins (gallotannins) in leaf (79.39%, 8.90 mg/g) and flower (60.00%, 3.50mg/g) while the stem was characterized by the predominance of flavonoid class (61.38%, 1.86 mg/g) due to the high presence of catechin (36.91%, 1.12 mg/g). Antioxidant activities of the essential oil and the methanolic extract from different myrtle parts were evaluated by using DPPH radical scavenging, beta-carotene-linoleic acid bleaching, reducing power and metal chelating activity assays. In all tests, methanolic extracts of different myrtle parts showed better antioxidant activity than essential oils.

Effect of Tunisian Capparis spinosa L. extract on melanogenesis in B16 murine melanoma cells.

The effect of Tunisian Capparis spinosa L. aromatic plant extract on melanogenesis regulation in B16 murine melanoma cells was investigated. B16 cells were treated with 0.0005, 0.005, and 0.05% (w/v) C. spinosa extract after which the melanin content and cell viability were measured. To clarify the mechanism behind melanogenesis regulation, the expression of tyrosinase was determined. Results showed that the extract had a significant stimulative effect on melanogenesis in B16 cells in a dose-dependent manner without cytotoxicity. Western blot analysis showed that expression of tyrosinase in cells treated with 0.03% (w/v) C. spinosa extract increased by 12.5- and 20-fold after 24 and 48 h of incubation, respectively, compared with untreated cells. HPLC analysis of the extract revealed the presence of 1% quercetin, a known melanogenesis stimulator, indicating that our findings may be attributed to quercetin; however, other compounds present in the extract may also have an effect on the overall ability of the extract to stimulate melanogenesis. We report here that Tunisian C. spinosa leaf extract can stimulate melanogenesis in a dose-dependent manner without cytotoxicity by increasing tyrosinase protein expression and has the potential to be used as a possible tanning agent or as a treatment for hair depigmentation.

Hair growth regulation by the extract of aromatic plant Erica multiflora.

Hair growth problems can affect human physical and mental health and are of particular relevance during the aging process. In an effort to resolve such problems, we attempted to find plants having hair growth regulation activity and thus collected plant extracts from Tunisia for bioprospecting purposes. Among them, we investigated the Erica multiflora extract to evaluate the hair growth promotion activity by using the MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) assay and cell cycle assay on human dermal papilla cells in vitro and an administration assay on mouse dorsal skin in vivo. The results showed that the Erica multiflora extract promotes dermal papilla cell growth and cell cycle with high activity, and induced hair growth in vivo by induction of anagen phase from telogen phase.

Antimelanogenesis effect of Tunisian herb Thymelaea hirsuta extract on B16 murine melanoma cells.

Skin pigmentation is the result of melanogenesis that occurs in melanocytes and/or melanoma cells. Although melanogenesis is necessary for the prevention of DNA damage and cancer caused by UV irradiation, excessive accumulation of melanin can also cause melanoma. Thus, we focused on the antimelanogenesis effect of an extract from Thymelaea hirsuta, a Tunisian herb. Murine melanoma B16 cells were treated with T. hirsuta extract, and then cell viability and synthesized melanin content were measured. We found that the T. hirsuta extract decreased the synthesized melanin content in B16 cells without cytotoxicity. Tyrosinase is a key enzyme of melanogenesis and extracellular signal-regulated kinase (ERK)-1/2 phosphorylation is known to be related to melanogenesis inhibition. To clarify its mechanism, we also determined ERK1/2 phosphorylation and tyrosinase expression level. ERK1/2 was immediately phosphorylated in cells just after treatment with the extract. The tyrosinase expression was inhibited after 24 h of stimulation with the extract. The T. hirsuta extract was fractionated, and we found that one fraction considerably decreased the melanin synthesis in B16 cells and that this fraction contains daphnanes as the main component. This indicates that our findings might be attributable to daphnanes.

Variations in phytosterol composition during the ripening of Tunisian safflower (Carthamus tinctorius L.) seeds.

Phytosterols are bioactive components of all vegetable foods. Their most studied and outstanding properties being their cholesterol-lowering activity. This property has led to the development of functional foods enriched with plant sterols. In the present study, total and conjugated sterols of Tunisian safflower seeds were investigated and their kinetic of accumulation was surveyed during the ripening stages. The results obtained revealed that total and individual phytosterols were actively accumulated during the first stages of seed development then their levels decreased until full maturity. In the other hand, the sterol profile was marked by the predominance of beta-sitosterol during all ripening stages. As for Free Sterols (FS) and Esterified Sterols (ES), they were the major components and were actively accumulated from the beginning of seed development. Nevertheless, their amounts decreased at the end of maturity while Steryl Glycosides (SG) and acylated sterylglycosides (ASG) were practically stable during the ripening of seeds.