Lorena Canuti

Biochemical Composition and Antioxidant Properties of Lavandula angustifolia Miller Essential Oil are Shielded by Propolis Against UV Radiations

UV radiations are principal causes of skin cancer and aging. Suntan creams were developed to protect epidermis and derma layers against photodegradation and photooxidation. The addition of antioxidant plant extracts (i.e. essential oil) to sunscreens is habitually performed, to increase their UV protective effects and to contrast pro‐radical and cytotoxic compounds present in these solutions. According to these observations, in the present work, the alteration of chemical composition and bioactive properties of Lavandula angustifolia Miller essential oil, exposed to UV light, was investigated. UV induced a significant deterioration of lavender oil biochemical profile. Moreover, the antioxidant activity of this solution, in in vitro tests and directly on B16‐F10 melanoma cells, greatly decreased after UV treatment. Our results also showed that essential oil was shielded from UV stress by propolis addition. Even after UV treatment, bee glue highly protected lavender oil secondary metabolites from degradation and also preserved their antiradical properties, both in in vitro antioxidant assays and in cell oxidative damage evaluations. This research proposed propolis as highly efficient UV protective and antiradical additive for sunscreens, cosmetics and alimentary or pharmaceutical products containing plant extracts.UV radiations are principal causes of skin cancer and aging. Suntan creams were developed to protect epidermis and derma layers against photodegradation and photooxidation. The addition of antioxidant plant extracts (i.e. essential oil) to sunscreens is habitually performed, to increase their UV protective effects and to contrast pro-radical and cytotoxic compounds present in these solutions. According to these observations, in the present work, the alteration of chemical composition and bioactive properties of Lavandula angustifolia Miller essential oil, exposed to UV light, was investigated. UV induced a significant deterioration of lavender oil biochemical profile. Moreover, the antioxidant activity of this solution, in in vitro tests and directly on B16-F10 melanoma cells, greatly decreased after UV treatment. Our results also showed that essential oil was shielded from UV stress by propolis addition. Even after UV treatment, bee glue highly protected lavender oil secondary metabolites from degradation and also preserved their antiradical properties, both in in vitro antioxidant assays and in cell oxidative damage evaluations. This research proposed propolis as highly efficient UV protective and antiradical additive for sunscreens, cosmetics and alimentary or pharmaceutical products containing plant extracts.

Tetracycline accumulates in Iberis sempervirens L. through apoplastic transport inducing oxidative stress and growth inhibition

Environmental antibiotic contamination is due mainly to improper and illegal disposal of these molecules that, yet pharmacologically active, are excreted by humans and animals. These compounds contaminate soil, water and plants. Many studies have reported the bioaccumulation of antibiotics in plants and their negative effects on photosynthesis, cell growth and oxidative balance. Therefore, the principal objective of this paper was the study of antibiotic accumulation sites in plants and its uptake modality. Iberis sempervirens L., grown in soil and in agar in the presence or absence of tetracycline, were used as a model system. Using confocal and transmission electron microscopy, we demonstrated that tetracycline was absorbed and propagated in plants through apoplastic transport and also accumulated in intercellular spaces. Tetracycline was rarely detected inside cells (in cytoplasm and mitochondria where, coherent to its pharmacological activity, it probably affected ribosomes), except in stomata. Moreover, we verified and clarified further the phytotoxic effects of tetracycline on plants. We observed that the antibiotic induced a large reduction in plant growth and development and inhibition of photosynthetic activity. As tetracycline may lead to oxidative stress in plants, plant cells tried to balance this disequilibrium by increasing the amount and activity of some endogenous enzyme antioxidant agents (superoxide dismutase 1 and catalase) and levels of antiradical secondary metabolites.

Identification of phenolic compounds from medicinal and melliferous plants and their cytotoxic activity in cancer cells

Abstract The aim of this work is to carry out a phytochemical analysis and biological screenings of vegetable extracts from Sida acuta and Malva sylvestris leaves, Castanea sativa and Eucalyptus camaldulensis pollen. Chemical analyses was focused on secondary metabolites, particularly phenolic compounds, which have several roles in the plant physiological processes and had demonstrated significant capacity in the prevention and care of human health diseases. Solid phase extraction (SPE) and analyses with liquid chromatography and mass spectrometry (HPLC-MS) allowed the identification of 5,7-dimethoxycoumarin, kaempferol, quercetin, genistein, apigenin and myricetin. Moreover, the M. sylvestris and S. acuta extracts demonstrated a cytotoxic activity on murine and human cancer cell lines by using a MTT assay.

Growth of Pseudomonas aeruginosa in zinc poor environments is promoted by a nicotianamine-related metallophore

Previous studies have suggested that P. aeruginosa possesses redundant zinc uptake systems. To identify uncharacterized zinc transporters, we analyzed the genome‐wide transcriptional responses of P. aeruginosa PA14 to zinc restriction. This approach led to the identification of an operon (zrmABCD) regulated by the zinc uptake regulator Zur, that encodes for a metallophore‐mediated zinc import system. This operon includes the genes for an uncharacterized TonB‐dependent Outer Membrane Protein (ZrmA) and for a putative nicotianamine synthase (ZrmB). The simultaneous inactivation of the ZnuABC transporter and of one of these two genes markedly decreases the ability of P. aeruginosa to grow in zinc‐poor media and compromises intracellular zinc accumulation. Our data demonstrate that ZrmB is involved in the synthesis of a metallophore which is released outside the cell and mediates zinc uptake through the ZrmA receptor. We also show that alterations in zinc homeostasis severely affect the ability of P. aeruginosa to cause acute lung and systemic infections in C57BL/6 mice, likely due to the involvement of zinc in the expression of several virulence traits. These findings disclose a hitherto unappreciated role of zinc in P. aeruginosa pathogenicity and reveal that this microorganism can obtain zinc through a strategy resembling siderophore‐mediated iron uptake.

Geographical, botanical and chemical profile of monofloral Italian honeys as food quality guarantee and territory brand

Abstract Bees represent an important element for the preservation of plant biodiversity. During their activities, bees cover a large area around the hive and their products strictly reflect the distinctive traits of these environments. In honey, the floral diversity of the areas inspected by bees is revealed by their pollen spectra. The present work characterized 460 Italian unifloral honeys by melissopalynological and physicochemical analyses. Obtained data contributed to: confirm the botanical identity of the samples; determine the plant biodiversity revealed in each honey; differentiate the samples of the same monofloral typology in relation to their geographical origin; and identify peculiar floristic associations in the Italian regions. Moreover, gas chromatographic analysis was carried out on rare unifloral honey to study their sugar profiles. This work provided the scientific base to create a valid tool able to assign a territorial brand and an authenticity guarantee to the honey, protecting the consumers from adulterations.

Trichomes in Camptotheca acuminata Decaisne (Nyssaceae): Morphology, distribution, structure, and secretion

Camptotheca acuminata is a main source of the anti-cancer drug camptothecin (CPT). In this species, several studies have observed non-glandular trichomes (NGTs) and glandular trichomes (GTs). It has been assumed that GTs contain CPT, yet this has not been proven and no information is available on the accumulation of other secondary metabolites. The objective of this study was to describe the morphology, distribution and structure of C. acuminata trichomes and to investigate the chemical nature of the substances secreted by GTs. Light and fluorescence microscope, scanning electron microscope (SEM) and transmission electron microscope (TEM) were used to determine the morphology, distribution and structure of GTs and NGTs. Thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC) analyses were carried out to confirm the presence of CPT in GTs, and histochemical tests were performed to investigate the presence of other secondary metabolites. C. acuminata possesses two types of GTs (GT1 and GT2), which differ in terms of their morphology, pattern of distribution and accumulated substances. The chemical analyses demonstrated that both GT1 and GT2 accumulate CPT. Histochemical analysis showed that phenols accumulate in the vacuole of GT2s. No isoprenoids were detected in GTs.

Nuclear shield: a multi-enzyme task-force for nucleus protection.

Background In eukaryotic cells the nuclear envelope isolates and protects DNA from molecules that could damage its structure or interfere with its processing. Moreover, selected protection enzymes and vitamins act as efficient guardians against toxic compounds both in the nucleoplasm and in the cytosol. The observation that a cytosolic detoxifying and antioxidant enzyme i.e. glutathione transferase is accumulated in the perinuclear region of the rat hepatocytes suggests that other unrecognized modalities of nuclear protection may exist. Here we show evidence for the existence of a safeguard enzyme machinery formed by an hyper-crowding of cationic enzymes and proteins encompassing the nuclear membrane and promoted by electrostatic interactions. Methodology/Principal Findings Electron spectroscopic imaging, zeta potential measurements, isoelectrofocusing, comet assay and mass spectrometry have been used to characterize this surprising structure that is present in the cells of all rat tissues examined (liver, kidney, heart, lung and brain), and that behaves as a “nuclear shield”. In hepatocytes, this hyper-crowding structure is about 300 nm thick, it is mainly formed by cationic enzymes and the local concentration of key protection enzymes, such as glutathione transferase, catalase and glutathione peroxidase is up to seven times higher than in the cytosol. The catalytic activity of these enzymes, when packed in the shield, is not modified and their relative concentrations vary remarkably in different tissues. Removal of this protective shield renders chromosomes more sensitive to damage by oxidative stress. Specific nuclear proteins anchored to the outer nuclear envelope are likely involved in the shield formation and stabilization. Conclusions/Significance The characterization of this previously unrecognized nuclear shield in different tissues opens a new interesting scenario for physiological and protection processes in eukaryotic cells. Selection and accumulation of protection enzymes near sensitive targets represents a new safeguard modality which deeply differs from the adaptive response which is based on expression of specific enzymes.

Nutritional and botanical interest of honey collected from protected natural areas

Abstract Due to numerous episodes of contamination through illegal beekeeping practices, the certification and re-evaluation of honey is required to guarantee a clean, natural and healthy product. The aim of this work was to demonstrate the good quality of honey produced in protected areas by demonstrating both its nutraceutical content and its role in monitoring the plant species present in protected areas. All the physico-chemical parameters necessary for defining the quality of honey (botanical origin; conductivity; diastase activity; pH; free, lactonic and total acidities; water content; fructose, glucose and sucrose content; hydroxymethylfurfural) and total flavonoid and phenolic contents were analyzed and evidenced its high quality. Moreover, melissopalynology allowed us to establish a floristic census within the nature reserves and parks.

Hydroalcoholic extract of Spartium junceum L. flowers inhibits growth and melanogenesis in B16-F10 cells by inducing senescence

BACKGROUND Ultraviolet light exposure generates, in human tissues, radical species, which represent the main cause of photo-aging, DNA damage and skin cancer onset. On the other hand, Mediterranean plants, being continuously subjected to high solar radiation levels, are naturally adapted to take on this type of abiotic stress, thanks to the production of antioxidant secondary metabolites. For these reasons, several plant extracts were documented to be excellent antineoplastic drugs. PURPOSE We investigated the potential antitumor activity of the flower extract obtained by Spartium junceum L., a Mediterranean shrub, correlating it with the plant metabolic profile. STUDY DESIGN After selecting the best extraction method to obtain as more secondary metabolites as possible from S. junceum flowers, we characterized the extract metabolic content. Then, by in vitro analyses, the antioxidant profile and the antineoplastic activity on B16-F10 murine melanoma cell of our extract were investigated. METHODS Spectrophotometric assays, HPLC-DAD and GC-MS analyses provided us information about flower extract composition and antioxidant activity. MTT assay and Trypan Blue exclusion test were performed to assess the extract toxicity and the viability, after treatments, of B16-F10 cancer cells and of C2C12 murine myoblasts. In vitro experiments (i.e. cytofluorimetry, protein analysis and qPCR) allowed us to analyze the effect of the plant extract on B16-F10 cell redox state, melanogenesis and cell cycle. Senescence induction was investigated by using a specific kit. RESULTS We observed that the hydroalcoholic extract of S. junceum flowers (HFE) strongly inhibited B16-F10 murine melanoma cell proliferation, while just a feeble effect was observed on C2C12 murine myoblasts. Moreover, we found that HFE exerted a pro-oxidant activity on melanoma cells, inhibited melanogenesis and caused cell cycle arrest in G2/M phase, inducing senescence. These anti-cancer properties of HFE could be related to the rich metabolic profile of the extract that we characterized by HPLC-DAD and GC-MS analyses. CONCLUSION This evidence suggests that S. junceum phytocomplex can be used as a selective, nontoxic, economic and easily available anticancer drug.

From Robinia pseudoacacia L. nectar to Acacia monofloral honey: biochemical changes and variation of biological properties: From Robinia pseudoacacia L. nectar to Acacia monofloral honey

BACKGROUND Robinia pseudoacacia L. nectar and its derivative monofloral honey were systematically compared in this study, to understand how much the starting solution reflected the final product, after re-elaboration by Apis mellifera ligustica Spinola. RESULTS Subjected to dehydration in the hive, nectar changed in its water and sugar content when transformed into honey, as physicochemical and gas chromatographic-mass spectrometric analyses revealed. Spectrophotometric measurements and characterization by high-performance liquid chromatography-diode array detection of 18 plant molecules demonstrated honey to be richer than nectar in secondary metabolites. For the first time, the hypothesis of the existence of a nectar redox cycle in R. pseudoacacia was reported, as previously described for Nicotiana sp., based on 1D-protein profiles, western blot analysis and detection of H2 O2 and ascorbate. The bioactivity of both matrices was also investigated. Antiradical in vitro tests showed that Acacia honey was more antioxidant than nectar, which was even able to induce oxidative stress directly in a eukaryotic cell system. Antimicrobial assays demonstrated that nectar was bacteriostatic, due to H2 O2 activity, whereas honey was even bactericidal. CONCLUSION All these data support the ecological role of nectar and honey in nature: protection of the gynoecium from pathogens and preservation from degradative processes, respectively.