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Euplotin C is a sesquiterpene of marine origin endowed with significant anti-microbial and anti-tumor properties. Despite the promising functional profile, its progress as a novel drug candidate has failed so far, due to its scarce solubility and poor stability in aqueous media, such as biological fluids. Therefore, overcoming these limits is an intriguing challenge for the scientific community. In this work, we synthesized β-cyclodextrin-based nanosponges and investigated their use as colloidal carriers for stably complex euplotin C. Results obtained proved the ability of the carrier to include the natural compound, showing remarkable values of both loading efficiency and capacity. Moreover, it also allowed us to preserve the chemical structure of the loaded compound, which was recovered unaltered once extracted from the complex. Therefore, the use of β-cyclodextrin-based nanosponges represents a viable option to vehiculate euplotin C, thus opening up its possible use as pharmacologically active compound. [ABSTRACT FROM AUTHOR]

Modern society is reviving the practice of aromatherapy, and lavender is reported being the most worldwide purchased plant for essential oil (EO) extraction. Since recent studies reported cognitive enhancing effects of lavender besides the hypno‐inducing effects, a literature review is needed. Considering EO quality and diffusion devices, we conducted a systematic review on the effects of lavender EO inhalation on arousal, attention and memory in healthy subjects. Starting from this new multidisciplinary perspective, cognitive effects were reviewed to link outcomes to effective and reproducible protocols. A systematic search on MEDLINE, ERIC, PsycInfo, Google Scholar, and Scopus databases using Cognitive Atlas and plant‐related keywords was conducted. Among the 1,203 articles yielded, 11 met eligibility criteria. Subjects administered with lavender EO displayed arousal decrease and sustained attention increase. Controversial results emerged regarding memory. Lack of EO quality assessment and protocols heterogeneity did not allow assessing whether different EO composition differentially modulates cognition and whether placebo effect can be discerned from EO effect itself. However, GABAergic pathway modulation exerted by linalool, a major lavender EO constituent, might explain cognitive functions empowerment. We speculate aromatherapy could be a burgeoning cognition enhancing tool, although further investigation is required to reach robust conclusions. [ABSTRACT FROM AUTHOR]

Introduction: Long-term stability of the treatment outcome of skeletal Class III malocclusion is always not achievable, therefore several prediction models of stability of orthopaedic treatment of the Class III malocclusion have been proposed. Aim: This cross-sectional study was aimed at the evaluation of the prognostic agreement and association between three different prediction models based on skeletal parameters recorded on lateral cephalograms. Materials and methods: A total of 75 subjects (34 females and 41 males, age range 7-11, mean age 9.1±1.2years) having skeletal Class III malocclusion were included in the study. Prediction models reported by Baccetti et al. 2004 (prediction model 1), Moon et al. 2005 (prediction model 2) and Yoshida et al. 2006 (prediction model 3), were considered. Percentage agreement and unweighted kappa coefficient evaluated the agreement between the prediction models in terms of stable or unstable cases. Multiple regressions were run to evaluate the association between the absolute scores obtained from each of the prediction model (prediction scores). Results: Percentage agreement ranged from 77.3% (models 2 and 3) to 89.3%, (models 1 and 2) and corresponding unweighted kappa coefficients ranged from 0.099 (models 1 and 3) to 0.205 (models 2 and 3). The prediction score from model 3 was significantly associated with those from both the other models; on the contrary, prediction scores from models 1 and 2 were not significantly associated. Conclusions: Aagreement in terms of stability or instability outcome derived from these models is only partial and not satisfactory, even though significant association between the prediction scores denotes a common biological meaning of the models. [ABSTRACT FROM AUTHOR]

Salvia wagneriana Polak is a tropical species native to Central America, well adapted to grow in the Mediterranean basin for garden decoration. Micropropagation has been assessed from axillary shoots of adult plants using a Murashige and Skoog basal medium, with the addition of 1.33-μM 6-benzylaminopurine for shoot proliferation; the subsequent rooting phase occurred in plant growth regulator-free medium. The plants were successfully acclimatised with high survival frequency. Hairy roots were induced after co-cultivation of leaf lamina and petiole fragments with Agrobacterium rhizogenes and confirmed by PCR. The establishment and proliferation of the selected HRD3 line were obtained in hormone-free liquid medium and the production of rosmarinic acid (RA) was evaluated after elicitation. The analysis of RA was performed by LC-ESI-DAD-MS in the hydroalcoholic extracts. The addition of casein hydrolysate increased the RA production, whereas no enrichment was observed after the elicitation with jasmonic acid. [ABSTRACT FROM AUTHOR]

Turkish Hypericum origanifolium Willd has been recently considered for its hypericin and phenolic content as a potential substitute of the well-established EU market position Hypericum perforatum . However, a complete phytochemical characterization of H. origanifolium has not carried out yet in order to assert its potential industrial value. The present paper represents the first attempt to define the aromatic fingerprint of a representative wild Turkish H. origanifolium Willd. population. In addition, its volatiles were studied as potential discriminating constituents between H. perforatum and H. origanifolium with a similar hypericin and phenolic composition. [ABSTRACT FROM AUTHOR]

The volatile constituents of the rare species ofHypericum,namelyHypericum pruinatum(as one population) andHypericum aviculariifoliumsubp.depilatumvar.depilatum(endemic, as two populations namely ‘Gümüş’ and ‘Yeniköy’) growing wild in the mountainous parts of Northern Turkey were studied for the first time. The essential oils (EOs) were extracted by hydrodistillation of the air-dried aerial parts and analysed by GC-FID and GC-MS. A total of 56, 49 and 50 EO components representing 98.2%, 96.9% and 99.4% of the total composition were identified respectively from one population forH. pruinatumand two populations forHypericum aviculariifoliumsubp.depilatumvar.depilatum. GC-MS profiles showed significant compositional variations not only between the two Turkish species, but also between the two populations of the same species highlighting the importance of genetic factors affecting secondary metabolite profile. [ABSTRACT FROM PUBLISHER]

Essential oils (EOs) of Achillea millefolium, Myrtus communis, Rosmarinus officinalis, Helichrysum italicum, Foeniculum vulgare and Lavandula angustifolia were analysed with GC–FID and GC–MS in order to define their aromatic profiles and then their toxicity and repellent activity against Sitophilus zeamais Motsch. (Coleoptera Dryophthoridae) with specific bioassays were evaluated. Results from topical applications on insects showed that all EOs had variable and significant insecticidal activity. Mortality rate never exceeded 76%. Results of repellency tests are indicated for M. communis and L. angustifolia EOs, displaying high repellent activity to S. zeamais adults. [ABSTRACT FROM PUBLISHER]

Context: In the past few years, an increasing interest in the volatile secondary metabolites of Hypericum perforatum L. (Guttiferae) has been arising. Objective: The present study is a contribution to better understand the relationship between the morphological variations and volatile composition during the phenological cycle. Materials and methods: Leaves at the stages of vegetative, floral budding, flowering and green capsule, buds, full opened flowers and green capsules were assayed for essential oil (EO) components by gas chromatography-flame ionization detector (GC-FID) and GC-mass spectrometry (MS). Results: Significant amounts of sesquiterpenes (oxygenated 26--50% and hydrocarbons 20--40%) and oxygenated hydrocarbons (13--38%) characterized the all analyzed samples showing peculiar fluctuations during the seven phenological stages. Although monoterpenes were present in much lower amounts (monoterpene hydrocarbons 0.4--6%; oxygenated monoterpenes 0.8--6%) they were considered also important discrimination for several stages. The green capsules and the full opened flowers collected at flowering stage were clearly distinguished in terms of EO compositions from the other samples. Discussion: For the first time, the EO composition of Turkish wild Hypericum perforatum was monitored by the hydrodistillation of different plant organs collected at different seven stages in order to point out the modification of target volatiles related to each phenological step. Conclusions: Based on the EO composition monitored during these seven morphological stages by GC-MS, principal component analysis and cluster analysis, significant metabolite modifications were observed during the phenological cycle which involved the levels of specific volatile target compounds belonging to the chemical classes of hydrocarbons, monoterpenes and sesquiterpenes. [ABSTRACT FROM AUTHOR]

Abstract: The volatile composition of ten fibre hemp (Cannabis sativa L.) varieties was investigated during two successive growing seasons under temperate climatic conditions in Central Italy. The freshly plant inflorescences were hydrodistilled and the essential oils (EOs) were characterized by GC–MS. In addition, the composition of the aroma emitted spontaneously from the freshly plant inflorescences were analysed by SPME-GC–MS. The EO yields of eight dioecious (Carmagnola, C.S., Red Petiole, Pop 1, Pop 2, Pop 3, Pop 4, Pop 5) and two monoecious (Codimono and Felina 34) cultivars ranged from 0.11 to 0.25% (w/w) and showed a significant production of α-pinene (3–20%), β-pinene (1–8%), E-ocimene (1–10%), myrcene (8–45%) and terpinolene (0.12–22%). The monoterpene composition was useful to distinguish the monoecious cultivars from the dioecious ones. β-Caryophyllene (7–28%), α-humulene (3–12%), and caryophyllene oxide (2–6%) were the main sesquiterpenes. Tetrahydrocannabinol (THC) was present in traces in the EOs of only two dioecious cultivars cultivated in 2005. Cannabinol (CBN) was not detected in the essential oils, while the no-hallucinogenous cannabidiol (CBD) was found as typical volatile constituent in several analysed cultivars. These findings were also confirmed by the headspace GC–MS analysis carried out on the same samples. The analysed EOs obtained from fibre hemp varieties cultivated in Central Italy were characterized by an interesting and specific terpene composition with a legal and safe cannabinoid content. They were obtained from freshly plant inflorescences, which usually represent a waste material from C. sativa L. fibre varieties. The present study strengths the hypothesis to grow hemp as a multi-use crop through a complete utilization of the plant material using inflorescences to produce essential oils as natural flavour and fragrance additives. [ABSTRACT FROM AUTHOR]

Laboratory bioassays on insecticidal activity of essential oils (EOs) extracted from six Mediterranean plants ( Achillea millefolium, Lavandula angustifolia, Helichrysum italicum, Foeniculum vulgare, Myrtus communis, and Rosmarinus officinalis) were carried out against the larvae of the Culicidae mosquito Aedes albopictus. The chemical composition of the six EOs was also investigated. Results from applications showed that all tested oils had insecticidal activity, with differences in mortality rates as a function of both oil and dosage. At the highest dosage (300 ppm), EOs from H. italicum, A. millefolium, and F. vulgare caused higher mortality than the other three oils, with mortality rates ranging from 98.3% to 100%. M. communis EO induced only 36.7% larval mortality at the highest dosage (300 ppm), a similar value to those recorded at the same dosage by using R. officinalis and L. angustifolia (51.7% and 55%, respectively). Identified compounds ranged from 91% to 99%. The analyzed EOs had higher content of monoterpenoids (80-99%) than sesquiterpenes (1-15%), and they can be categorized into three groups on the basis of their composition. Few EOs showed the hydrocarbon sesquiterpenes, and these volatile compounds were generally predominant in comparison with the oxygenated forms, which were detected in lower quantities only in H. italicum (1.80%) and in M. communis (1%). [ABSTRACT FROM AUTHOR]

Context: Hypericum perforatum L. (Guttiferae) contains many bioactive secondary metabolites including hypericins, hyperforins, and essential oil. Objective: The present study was conducted to determine the variation in composition of essential oil in H. perforatum accessions from Turkey. Material and methods: At full flowering, aerial parts of 30 plants were collected from 10 sites of northern Turkey and assayed for essential oil components by GC-FID and GC-MS. Results: The chemical analysis revealed that the main constituents of the all analyzed samples were hydrocarbon and oxygenated sesquiterpenes such as β-caryophyllene (4.08–5.93%), γ-muurolene (5.00–9.56%), β-selinene (5.08–19.63%), α-selinene (4.12–10.42%), -cadinene (3.02–4.94%), spathulenol (2.34–5.14%), and caryophyllene oxide (6.01–12.18%). Monoterpenes, both hydrocarbon and oxygenated, were represented by scarce amounts of α- and β-pinene, myrcene, linalool, cis- and trans-linalool oxide, and α-terpineol. Principal component analysis was also carried out and, according to the results, the first nine principal components were found to represent 100% of the observed variation. Discussion: The chemical variation among the populations is discussed as the possible result of different genetic and environmental factors. Conclusions: The wild populations examined here are potentially important sources for breeding and improvement of the cultivated varieties. [ABSTRACT FROM AUTHOR]

A laccase-based biosensor was developed by specific enzyme adsorption on screen-printed working electrodes of DROPSENS cells, and stabilized with Nafion 0.1% membrane. The electrode was characterized with respect to response time, sensitivity, linear range, detection limit, pH dependence, interferences, and long-term stability. The tested substrates were catechol, rosmarinic acid, caffeic acid, chlorogenic acid, and gallic acid. The optimized biosensor proved the following characteristic performances: the apparent Michaelis Menten calculated considering rosmarinic acid substrate 8.3 × 10-6 mol L-1 (r = 0.995, n = 6); the dynamic range of biosensor response for rosmarinic acid 7 × 10-7 - 1.5 × 10-6 mol L-1; the detection limit for rosmarinic acid 1.19 × 10-7 mol L-1 (RSD = 1.08%, n = 3). It was noticed that the biosensor reaches systematically 90% to 94.3% from the response obtained by LC-DAD-ESI-MS for real samples. [ABSTRACT FROM AUTHOR]

Cytisus aeolicus Guss. ex Lindl. ( Fabaceae family, subfamily Faboideae) is an endangered endemic species of the Aeolian Islands, Sicily. In vitro multiplication of C. aeolicus shoots was described in this work and cell cultures were established from cotyledons and hypocotyls to investigate their potential production of isoflavones. Aseptically germinated seeds, cultivated on LS modified basal medium, gave the initial explants used both to induce axillary propagation and callus cultures. The LS (Linsmaier and Skoog) basal medium, supplemented with 0.1 mg L−1 of 6-benzylaminopurine were used to induce axillary propagation. The callus induction was performed using the basal medium added with 5 mg L−1 2,4-dichlorophenoxy acetic acid and 5 mg L−1 kinetin (control medium). Basal medium was also added with 2000 mg L−1 casein hydrolysate (CH) or 900 mg L−1 myo-inositol (MI). C. aeolicus callus cultures on CH and MI media produced an unique compound, the isoflavone genistein 7-O-ß-D-glucopyranoside (genistin), which has not previously been isolated from wild plants. Callus cultures grown on the medium containing myo-inositol produced the greatest amount of genistin. C. aeolicus tissue culture procedures could provide suitable plant material both for germplasm preservation (by micropropagation) and for biotechnological selective isoflavone production (by callus culture). [ABSTRACT FROM AUTHOR]

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A series of task-specific ionic liquids (TSILs) based on glycerylimidazolium cations have been prepared by reaction of 1-chloropropanediol, a compound obtainable from glycerol (a widely available and in-expensive waste product), with the appropriate base (1-H-imidazole, 1,2-dimethylimidazole and 1-methyl-1-H-imidazole). The reaction of 3-(1H-imidazol-1-yl)propane-1,2-diol with chloroalkanes, bromoalkanes and alkyl mesylates gave the corresponding salts which were characterized. The possibility to use these ILs in palladium catalyzed reactions was evaluated, evidencing good catalyst stability and a high recyclability. [ABSTRACT FROM AUTHOR]

Six flavonol glycosides were isolated and identified from the aerial parts of Anthyllis barba-jovis L. (Leguminosae), together with two coumarins and D-pinitol, a taxonomic marker of Leguminosae family. The structural elucidation of all compounds was based on their 1H- and 13C-NMR spectral data and bidimensional experiments. The total flavonoid content was also determined, according to the method described in the Italian pharmacopoeia. In order to complete the phytochemical investigation on A. barba-jovis, the essential oils from flowers and seeds were obtained by hydrodistillation and analysed by GC-MS; fresh flowers were analysed also directly by solid phase micro extraction (SPME). [ABSTRACT FROM AUTHOR]

Smyrnium olusatrum L. is a biennal plant growing in western and southern Europe. Several phytochemical studies have identified flavonoids, phenolic acids and sesquiterpene lactones as main constituents. In this work the root, stem and leaf essential oils of Smyrnium olusatrum L. (Umbelliferae) collected in Urbino (Marche, Italy) were studied by GC and GC-MS. Myrcene (1, 13 and 14%), β-phellandrene (34, 17 and 9%), β-caryophyllene (1, 3 and 12%), furanodiene (19, 8 and 17%), curzerene (13, 6 and 12%), germacrene B (7, 14 and 10%) and germacrone (8, 7 and 8%) were identified as the major components in the root, stem and leaf oils, respectively. SPME analysis of fresh material of the same different organs showed β-myrcene (3, 21 and 36%), β-phellandrene (25, 34 and 12%), β-caryophyllene (1, 7 and 22%) and furanodiene (4, 7 and 7%) as typical components in the headspace of fresh roots, stems and leaves, respectively. Copyright © 2004 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

The essential oil and solid phase micro extraction (SPME) samples of Bupleurum fruticosum micropropagated plants were analysed by GC and GC–MS and compared with those obtained from the leaves and the stems of field-grown parent plants. The main constituents of the essential oil of the micropropagated plants were β-phellandrene (61%), sabinene (13%), terpinen-4-ol, tricyclene and bicyclogermacrene (3%). Regarding to the field-grown B. fruticosum plants, the leaf essential oil showed the same major components detected in the micropropagated plants, while the stem oil showed γ-terpinene (50%) and α-phellandrene (18%) as the most important constituents. Furthermore, SPME analyses were carried out in order to show for the first time a complete investigation on the volatile organic constituents of the micropopagated and field-grown B. fruticosum parent plants. [Copyright &y& Elsevier]

From the aerial parts of Astragalus verrucosus Moris, a novel cycloartane-type triterpene glycoside, named astraverrucin VII, was isolated along with cycloaraloside D (peregrinoside II) and cycloaraloside C (astrailienin A). The flavonoid composition was investigated for the first time, and fifteen known flavonoids were isolated and identified. All structural elucidation were performed by spectral means. The chemiotaxonomic importance of these findings is discussed. [ABSTRACT FROM AUTHOR]

Six weld genotypes (Reseda luteola L.) were evaluated for their agronomic characteristics in a 4-year field study carried out under rainfed condition in Central Italy in order to point out productive potential and the best harvest time to maximise yield of dye. Good agronomic performances were observed with an overall dry yield of about 40 g per plant and an estimated dry yield per hectare of 2.8 tons. Differences were found among the six accessions and some of them withstanded well to drought conditions experienced in drier growing seasons. The harvest of plants during flowering or during beginning of fruit ripening did not affect overall dry yield (40 and 45 g per plant at flowering and fruit maturity, respectively). The luteolin amount (expressed in mg g−1 dry weight) was affected by climatic conditions as experienced in two different growing seasons (1993 and 1994) and by the plant phenological stages at harvest being luteolin most abundant in the dryest year and in plants harvested during flowering. Genetic variability existed for the luteolin content in leaves plus reproductive structures (inflorescences and fruits) that varied from 13.6 to 28.7 mg g−1 dry weight in the six accessions tested in 1994. The dry powder from leaves and reproductive structures was used in the dyeing of cotton, wool and silk yarns. Good and bright yellow colours were observed using a ratio 30:100 (weight/weight) of weld on the textile materials. Separated weld organs showed different dyeing capacities being the leaves, inflorescences and fruits the most effective ones. Weld showed a progressive decrease in resistance to fading going from cotton, to the other kind of yarns. Finally, all dyed specimens exhibited good resistance to light and wash fastness making the use of weld a viable alternative to synthetic yellow dyes. [Copyright &y& Elsevier]

The antimicrobial and antifungal properties of several crude extracts and pure saponins, astraverrucins I–VI, from the aerial parts of Astragalus verrucosus were investigated by the disc diffusion method. Antimycotic activity was observed on more polar extracts, but only one saponin showed an appreciable activity. [Copyright &y& Elsevier]

Glioblastoma multiforme (GBM) is an aggressive brain tumour with high resistance to radio- and chemotherapy. As such, increasing attention has focused on developing new therapeutic strategies to improve treatment responses. Recently, attention has been shifted to natural compounds that are able to halt tumour development. Among them, carnosol (CAR), a phenolic diterpene present in rosemary, has become a promising molecule that is able to prevent certain types of solid cancer. However, no data are available on the effects of CAR in GBM. Here, CAR activity decreased the proliferation of different human glioblastoma cell lines, particularly cells that express wild type p53. The p53 pathway is involved in the control of apoptosis and is often impaired in GBM. Notably, CAR, through the dissociation of p53 from its endogenous inhibitor MDM2, was able to increase the intracellular p53 levels in GBM cells. Accordingly, functional reactivation of p53 was demonstrated by the stimulation of p53 target genes’ transcription, the induction of apoptosis and cell cycle blockade. Most importantly, CAR produced synergistic effects with temozolomide (TMZ) and reduced the restoration of the tumour cells’ proliferation after drug removal. Thus, for the first time, these data highlighted the potential use of the diterpene in the sensitization of GBM cells to chemotherapy through a direct re-activation of p53 pathway. Furthermore, progress has been made in delineating the biochemical mechanisms underlying the pro-apoptotic effects of this molecule. [ABSTRACT FROM AUTHOR]

The EU project Nutra-Snack established an integrated scientific and technological framework to introduce conventional and new biotechnologies into the nutraceutical industry. The purpose was to promote novel ready-to-eat snacks enriched with natural antioxidant/radical-scavenging compounds. Enhancement of valuable compounds production in plant cells and in vitro cultures were achieved by different elicitation strategies, and exhaustive metabolites profiling obtained by high-resolution analytical techniques. Finally, innovative enzyme-based biosensors were developed for food products functional activity and safety properties assessment. This review provides an overview of the technologies selected, integrated and tested in NUTRA-SNACK and recent literature strengthening integrated approaches in modern food process manufacturing. [ABSTRACT FROM AUTHOR]

Curcuma longa L. (turmeric) is one of the most important spice and safe food additives. Its main constituents, curcuminoids, showed anti-inflammatory, antitumor and antioxidant effects. In the present work, an in vitro propagation method was developed to achieve selected plant organs with quantified curcuminoid content. In vitro plants were obtained from sprouting buds as primary explants. The major curcuminoid constituents, such as curcumin (CUR), demethoxycurcumin (DEM), and bis-demethoxycurcumin (bis-DEM) were examined in different organs by LC-DAD-ESI-MS. A significant production of curcumin (more than 260 microg g(-1) fresh weight) was obtained from in vitro microrhizomes, especially grown in a Murashige and Skoog medium (MS) supplemented with kinetin (0.1 mg L((-1)), alpha-naphthaleneacetic acid (NAA, 1 mg L(-1)), sucrose (6%), agar (5%) and activated charcoal (0.1%). The analyzed microrhizomes showed reduced amounts of DEM and bis-DEM in comparison with CUR levels. In addition a shoot culture line was suitable to biosynthesize curcuminoids, in a ratio very similar to that identified in the fresh rhizomes of parent plants. This study represents the first direct quantification of curcuminoids in turmeric in vitro shoots and microrhizomes to be used in dietary supplements.

The essential oils (EOs) and static headspaces (HSs) of in vitro plantlets and callus of Mentha x piperita were characterized by GC-MS analysis. Leaves were used as explants to induce in vitro plant material. The EO yields of the in vitro biomass were much lower (0.1% v/w) than those of the parent plants (2% v/w). Many typical mint volatiles were emitted by the in vitro production, but the callus and in vitro plantelet EOs were characterized by the lack of both pulegone and menthofuran. This was an important difference between in vitro and in vivo plant material as huge amounts of pulegone and menthofuran may jeopardise the safety of mint essential oil. Regarding the other characteristic volatiles, menthone was present in reduced amounts (2%) in the in vitro plantlets and was not detected in the callus, even if it represented the main constituent of the stem and leaf EOs obtained from the cultivated mint (26% leaves; 33% stems). The M. piperita callus was characterized by menthol (9%) and menthone (2%), while the in vitro plantlet EO showed lower amounts of both these compounds in favour of piperitenone oxide (45%). Therefore, the established callus and in vitro plantlets showed peculiar aromatic profiles characterized by the lack of pulegone and menthofuran which have to be monitored in the mint oil for their toxicity.

In vitro plantlets and callus of M. longifolia were established and their volatile constituents characterized by GC-MS analysis of their headspaces (HSs) and essential oils (EOs). Significant quali-quantitative differences were found in the aromatic fingerprints in comparison with the M. longifolia parent plants. In fact, limonene and carvone were the main constituents in the EOs of the mother plants, while the aroma of the in vitro plant material were especially enriched in oxygenated terpenes. In particular, huge amounts of piperitenone and piperitenone oxide (75 %) were found for in vitro plantlets, while trans-carvone oxide (19 %) and trans-piperitone epoxide (9 %) were found in callus EO. However, the established in vitro plant material showed lack of pulegone and menthofurane, thus preserving an important feature observed in the volatile fingerprint of the parent plants. In fact, because of their well-known toxicity significant amounts of pulegone and menthofurane may compromise the safety using of mint essential oil. Therefore the in vitro M. longifolia plantlets and callus may be regarded as a potential source of a safe flavouring agent.

Context: Hypericum perforatum L. (Guttiferae) contains many bioactive secondary metabolites including hypericins, hyperforins, and essential oil., Objective: The present study was conducted to determine the variation in composition of essential oil in H. perforatum accessions from Turkey., Material and Methods: At full flowering, aerial parts of 30 plants were collected from 10 sites of northern Turkey and assayed for essential oil components by GC-FID and GC-MS., Results: The chemical analysis revealed that the main constituents of the all analyzed samples were hydrocarbon and oxygenated sesquiterpenes such as beta-caryophyllene (4.08-5.93%), gamma-muurolene (5.00-9.56%), beta-selinene (5.08-19.63%), alpha-selinene (4.12-10.42%), d-cadinene (3.02-4.94%), spathulenol (2.34-5.14%), and caryophyllene oxide (6.01-12.18%). Monoterpenes, both hydrocarbon and oxygenated, were represented by scarce amounts of alpha- and beta-pinene, myrcene, linalool, cis- and trans-linalool oxide, and alpha-terpineol. Principal component analysis was also carried out and, according to the results, the first nine principal components were found to represent 100% of the observed variation., Discussion: The chemical variation among the populations is discussed as the possible result of different genetic and environmental factors., Conclusions: The wild populations examined here are potentially important sources for breeding and improvement of the cultivated varieties.

The aromatic profiles of five commercial thyme cultivars (T. vulgaris 'Silver Poise', T. vulgaris 'Erectus', T. vulgaris 'Faustini', T. x citriodorus 'Anderson's Gold', and T. x citriodorus 'Silver Queen'), cultivated in Italy, were defined both by their static headspaces (HS) and essential oils (EOs). In addition, a botanical garden sample of T. vulgaris was considered as reference material to evaluate the morphological and phytochemical differences from the selected market samples. Extractions of the volatile constituents of the different plant material were carried out by SPME (static headspace, HS) and hydrodistillation (HD) processes. GC-MS analysis provided the separation and identification of approximately 70 components in the HS samples and 50 in the hydrodistilled essential oils, accounting for more than 95% of the total. The typical main constituents of T. vulgaris were detected in all the EO samples, although qualitative and quantitative differences were found among the selected ornamental Thymus varieties. Thymol (50-55%) was the marker constituent for the three T. vulgaris cultivars, while geraniol (61-67%) characterized the essential oils of the two T. x citriodorus varieties. In all the analyzed essential oils, non-oxygenated (16-79%) and oxygenated (5-26%) monoterpenes were the typical volatile constituents. Specific target compounds (thymol, geraniol and 6-methyl-5-hepten-2-one) were selected to characterize the five thyme cultivars that are considered at present only as ornamental plants. A comparative evaluation of their EO quality was carried out in order to propose them as alternative sources of Italian raw plant material for industrial production.

The recent biotechnology boom has triggered increased interest in plant cell cultures, since a number of firms and academic institutions investigated intensively to rise the production of very promising bioactive compounds. In alternative to wild collection or plant cultivation, the production of useful and valuable secondary metabolites in large bioreactors is an attractive proposal; it should contribute significantly to future attempts to preserve global biodiversity and alleviate associated ecological problems. The advantages of such processes include the controlled production according to demand and a reduced man work requirement. Plant cells have been grown in different shape bioreactors, however, there are a variety of problems to be solved before this technology can be adopted on a wide scale for the production of useful plant secondary metabolites. There are different factors affecting the culture growth and secondary metabolite production in bioreactors: the gaseous atmosphere, oxygen supply and CO2 exchange, pH, minerals, carbohydrates, growth regulators, the liquid medium rheology and cell density. Moreover agitation systems and sterilization conditions may negatively influence the whole process. Many types ofbioreactors have been successfully used for cultivating transformed root cultures, depending on both different aeration system and nutrient supply. Several examples of medicinal and aromatic plant cultures were here summarized for the scale up cultivation in bioreactors.

Hairy roots (HRs) are differentiated cultures of transformed roots generated by the infection of wounded higher plants with Agrobacterium rhizogenes. This pathogen causes the HR disease leading to the neoplastic growth of roots that are characterized by high growth rate in hormone free media and genetic stability. HRs produce the same phytochemicals pattern of the corresponding wild type organ. High stability and productivity features allow the exploitation of HRs as valuable biotechnological tool for the production of plant secondary metabolites. In addition, several elicitation methods can be used to further enhance their accumulation in both small and large scale production. However, in the latter case, cultivation in bioreactors should be still optimized. HRs can be also utilised as biological farm for the production of recombinant proteins, hence holding additional potential for industrial use. HR technology has been strongly improved by increased knowledge of molecular mechanisms underlying their development. The present review summarizes updated aspects of the hairy root induction, genetics and metabolite production.

The production of plant secondary metabolites by in vitro culture is one of the most challenging and thrilling field of recent scientific researches. In the few last years, pharmaceutical and food industry demand in phytochemicals has increased steadily. Therefore, the establishment of in vitro plant protocols has to be monitored by phytochemical investigation of their selected extracts in order to supply standardized raw material. In this chapter, the advantages and disadvantages of some modem techniques have been described for the sampling, extraction and analysis ofthe invitro plants and derivatives. Depending on the volatile or nonvolatile substances produced by in vitro plant raw material, different kinds of laboratory facilities are needed for the extraction and quali-quantitative analysis. Recent extraction technology such as accelerated solvent extraction or microwave assisted extraction in combination with hyphenated techniques such as gas chromathography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) represent a modern approach to perform fast and reproducible analytical methods for the quality control of secondary metabolite production in 'in vitro' plant material.

The EtOAc and n-BuOH extracts of Inga fendleriana inhibited Gram-positive, but not Gram-negative bacteria; a narrow spectrum of activity against Staphylococcus epidermidis was detected. The MIC values of the extracts ranged from 125 to 850 microg/mL. Quercetin 3-methylether, myricetin 3-O-rhamnoside and tricetin showed antibacterial activity against the same bacterial strains with MICs in the range from 31 to 250 microg/mL. In time-kill kinetic studies, the flavonoids showed bactericidal effects at the concentrations corresponding to four times the MICs.

A wild strain of Agrobacterium rhizogenes was used to regenerate twelve in vitro plant lines from different hairy roots of H. perforatum (St. John's Wort). The production of the main bioactive constituents was observed even though their yields varied in the different plant lines. Two lines were selected for the hyperoside production (4.9-4.6 mg/gdw) while nine were characterized by significant yields of chlorogenic acid (ranged from 0.47 to 1.09 mg/gdw). Furthermore, one out of twelve lines showed a 10-fold higher hypericin content (0.25 mg/gdw) than that reported for the in vitro shoots in the literature. Morphological and phytochemical features were determined in order to select H. perforatum genotypes enriched in valuable bioactive compounds.