Allium sativum L. var. Voghiera Reduces Aflatoxin B1 Bioaccessibility and Cytotoxicity In Vitro.

The present work focuses on the evaluation of AFB1's bioaccessibility and cytotoxicity in vitro using bread (naturally contaminated) enriched or not enriched with fresh Voghiera garlic (2%). Two different experiments were carried out: experiment 1 (E1), with low-AFB1-concentration breads (1.6-1.7 mg/kg); and experiment 2 (E2), with high-AFB1-concentration breads (96.4-102.7 mg/kg). Eight breads were prepared, four for E1 (experiment 1) and another four for E2 (experiment 2), with each experiment having a control group (C), a garlic-enriched group (2%) (G), an AFB1 group (A), and an AFB1 + garlic group (A + G). Simulated digestion was performed on each type of bread, and gastric and intestinal digests were obtained. AFB1 content in flours, baked bread, and gastric and intestinal digests was measured by High-Performance Liquid Chromatography coupled to Fluorescence Detection. The results demonstrate dose-dependent AFB1 bioaccessibility and that the presence of garlic contributed to its reduction in both doses (7-8%). Moreover, garlic's presence in AFB1-contaminated bread increased cell viability (9-18%) in differentiated Caco-2 cells and mitigated the arrest of S and G2/M phases provoked by AFB1 on Jurkat T cells and reduced apoptosis/necrosis, cellular reactive oxygen species (ROS), and mitochondrial ROS by 16%, 71%, and 24% respectively. The inclusion of garlic as a functional ingredient helped relieve the presence and effects of AFB1.

Influence of Post-Harvest 1-Methylcyclopropene (1-MCP) Treatment and Refrigeration on Chemical Composition, Phenolic Profile and Antioxidant Modifications during Storage of Abate Fétel Pears.

'Abate Fétel', a winter cultivar, is the most important pear cultivar in Italy; its fruits are appreciated by consumers for their aroma, texture and balanced sweet and sour taste. Maintaining high-quality characteristics to prolong the shelf-life of fruit and preserve the sensory and nutritional quality is a priority for the food industry. The aim of our study was to test the effectiveness of 1-methylcyclopropene (1-MCP) and cold storage in prolonging the shelf-life of these fruits, which were harvested at maturity at two different times. This work focused on the effects of different storage treatments and two ripening times on (i) the chemical composition of Abate Fétel pulp fruits to preserve their sweet taste and aroma and (ii) the phenolic profile composition and antioxidant activity of the peel, which is naturally rich in phytochemicals and important for the fruit's shelf-life and in the functional food industry for its high nutritional value. Abate Fétel fruits were harvested at the optimal commercial maturity stage, first on 15 September, having been treated with 1-MCP and stored for 2 months at cold temperatures; the other fruits were harvested at the end of September and stored in a cold cell for 2 months. The fruit pulp was tested for glucose and fructose, pH, acidity and organic acids (malic, citric, fumaric and shikimic), phenolic content and phenolic compounds (chlorogenic and caffeic acids, rutin, hyperoside, kaempferol-3-rutinoside and isoquercitrin), and the antioxidant activities in the fruit peels were measured. Treating the fruits with 1-MCP better preserved the phytochemical compounds compared to simple refrigeration, preserving the fruit's quality and prolonging its shelf-life. All the treatments help to maintain the glucose and fructose content and the acidity, preserving the aroma and organoleptic characteristics.

Nutrients and Main Secondary Metabolites Characterizing Extracts and Essential Oil from Fruits of Ammodaucus leucotrichus Coss. & Dur. (Western Sahara).

The ethnobotany of the Sahrawi people considers various species of plants and crude drugs as food, cooking spices and traditional health remedies. From among these, the fruits of Ammodaucus leucotrichus Coss. & Dur. (Apiaceae), known as Saharan cumin, were chosen for our research. The present paper reports a proximate composition and mineral element analysis of various samples of A. leucotrichus fruits, collected during the balsamic period (full fruiting) from plants grown in Bir Lehlu (Western Sahara) and purchased in a local market (Tindouf). These analyses pointed out interesting nutritional values of the crude drug. Decoction and alcoholic extract, analyzed by HPLC-DAD, evidenced ammolactone-A and R-perillaldehyde as the two main isolated constituents, particularly in the ethanolic extracts (ammolactone-A, market sample: 51.71 ± 0.39 mg/g dry extract; wild sample: 111.60 ± 1.80 mg/g dry extract; R-perillaldehyde, market sample: 145.95 ± 0.35 mg/g dry extract; wild sample: 221.40 ± 0.30 mg/g dry extract). The essential oils, obtained through hydrodistillation, were characterized by GC-MS and evidenced R-perillaldehyde (market sample: 53.21 ± 1.52%; wild sample: 74.01 ± 1.75%) and limonene (market sample: 35.15 ± 1.68%; wild sample: 19.90 ± 1.86%) as the most abundant compounds. The R configuration of perillaldehyde was ascertained and a complete description of the 1H and 13C NMR spectra of ammolactone-A was performed.

Antioxidant Activity of Resveratrol Diastereomeric Forms Assayed in Fluorescent-Engineered Human Keratinocytes.

Resveratrol is a powerful antioxidant molecule. In the human diet, its most important source is in Vitis vinifera grape peel and leaves. Resveratrol exists in two isoforms, cis- and trans. The diastereomeric forms of many drugs have been reported as affecting their activity. The aim of this study was to set up a cellular model to investigate how far resveratrol could counteract cytotoxicity in an oxidant agent. For this purpose, a keratinocyte cell line, which was genetically engineered with jelly fish green fluorescent protein, was treated with the free radical promoter Cumene hydroperoxide. The antioxidant activity of the trans-resveratrol and its diastereomeric mixture was evaluated indirectly in these treated fluorescent-engineered keratinocytes by analyzing the cell number and cell proliferation index. Our results demonstrate that cells, which were pre-incubated with resveratrol, reverted the oxidative damage progression induced by this free radical agent. In conclusion, fluorescent-engineered human keratinocytes represent a rapid and low-cost cellular model to determine cell numbers by studying emitted fluorescence. Comparative studies carried out with fluorescent keratinocytes indicate that trans-resveratrol is more efficient than diastereomeric mixtures in protecting cells from the oxidative stress.

Characterization of Neochloris oleoabundans under Different Cultivation Modes and First Results on Bioactivity of Its Extracts against HCoV-229E Virus.

Microalgae are proposed in several biotechnological fields because of their ability to produce biomass enriched in high-value compounds according to cultivation conditions. Regarding the health sector, an emerging area focuses on natural products exploitable against viruses. This work deals with the characterization of the green microalga Neochloris oleoabundans cultivated under autotrophic and mixotrophic conditions as a source of whole aqueous extracts, tested as antivirals against HCoV-229E (Coronaviridae family). Glucose was employed for mixotrophic cultures. Growth and maximum quantum yield of photosystem II were monitored for both cultivations. Algae extracts for antiviral tests were prepared using cultures harvested at the early stationary phase of growth. Biochemical and morphological analyses of algae indicated a different content of the most important classes of bioactive compounds with antiviral properties (lipids, exo-polysaccharides, and total phenolics, proteins and pigments). To clarify which phase of HCoV-229E infection on MRC-5 fibroblast cells was affected by N. oleoabundans extracts, four conditions were tested. Extracts gave excellent results, mainly against the first steps of virus infection. Notwithstanding the biochemical profile of algae/extracts deserves further investigation, the antiviral effect may have been mainly promoted by the combination of proteins/pigments/phenolics for the extract derived from autotrophic cultures and of proteins/acidic exo-polysaccharides/lipids in the case of mixotrophic ones.

Anticancer Activity of Aqueous Extracts from Asparagus officinalis L. Byproduct on Breast Cancer Cells.

Cultivation of asparagus (Asparagus officinalis L.; Asp) for food and medicinal use has taken place since the early Roman Empire. Today, Asp represents a worldwide diffuse perennial crop. Lower portions of the spears represent a food industry waste product that can be used to extract bioactive molecules. In this study, aqueous extracts derived from the non-edible portion of the plant (hard stem) were prepared and characterized for chemical content. Furthermore, the biocompatibility and bioactivity of Asp aqueous extracts were assessed in vitro on normal fibroblasts and on breast cancer cell lines. Results showed no interference with fibroblast viability, while a remarkable cytostatic concentration-dependent activity, with significant G1/S cell cycle arrest, was specifically observed in breast cancer cells without apoptosis induction. Asp extracts were also shown to significantly inhibit cell migration. Further analyses showed that Asp extracts were characterized by specific pro-oxidant activity against tumoral cells, and, importantly, that their combination with menadione resulted in a significant enhancement of oxidants production with respect to menadione alone in breast cancer cells but not in normal cells. This selectivity of action on tumoral cells, together with the easiness of their preparation, makes the aqueous Asp extracts very attractive for further investigation in breast cancer research, particularly to investigate their role as possible co-adjuvant agents of clinical drug therapies.

Ethanol-based garlic extract prevents malignant evolution of non-invasive breast tumor cells induced by moderate hypoxia.

BACKGROUND: In breast cancer, low oxygen availability is associated with a more aggressive phenotype and with malignant evolution of non-invasive cells. Natural compounds have long attracted attention in cancer treatment, and in recent years garlic (Allium sativum) organosulfur derivatives have been shown to negatively affect growth and invasion of tumor cells. METHODS: Homemade ethanol-based garlic extract (GE) was administered to MCF7 and MCF10DCIS breast tumor cell lines grown under moderate hypoxia. Cell cycle, epithelial-to-mesenchymal transition and cancer stem cell markers were evaluated. RESULTS: We revealed that, in the non-invasive MCF10DCIS cells but not in the post-EMT MCF7 cells, low oxygen availability induced the decrease of E-cadherin and the increase of vimentin and motility, that were prevented by GE administration. In both cell lines, treatment with GE counteracted the up-modulation of CD133 positive cells induced by hypoxia. CONCLUSIONS: Overall, our data firstly revealed anti-cancer properties of garlic in non-invasive breast cancer cells. In particular, they demonstrated a protective role of this natural product against the hypoxia-induced increase of molecules that play crucial roles in tumor evolution, suggesting that garlic derivatives can be considered in new approaches for preventing progression of breast tumors from non-invasive to infiltrating lesions.

Nutrient Composition and Antioxidant Performances of Bread-Making Products Enriched with Stinging Nettle (Urtica dioica) Leaves.

Stinging nettle (Urtica dioica) is an edible plant, well-known for its nutritional and nutraceutical properties. Stinging nettle leaves are typically rich in fibers, minerals and vitamins, as well as antioxidant compounds, i.e., polyphenols and carotenoids. Due to these reasons, since ancient times stinging nettle has been widely used in Italy as an ingredient in foods and beverages as a therapeutic agent. This work provides an investigation focused on bread enrichment with nettle leaves and the improvement of bread proximate composition in minerals, fibers and antioxidant compounds during product preparation. The comparison between plain and nettle enriched white bread shows a significant increase in fibers and nutrients, i.e., calcium and copper levels. Nettle enrichment also provides an increase in lutein and ß-carotene, as well as in total phenols and antioxidant activity. These last two nutritional elements are remarkably high in enriched bread and it has been found that phenolic concentration increases during breadmaking steps, from kneading to primary dough fermentation and from secondary fermentation of shaped loaves to baking.

Preliminary multi analytical approach to address geographic traceability at the intraspecific level in Scombridae family.

Globalization of seafood product marketing caused the increase of request of an effective fish traceability that enhances the consumer confidence in food safety. In this study, an integrated multi analytical approach based on two different and independent analytical techniques (carbon and nitrogen stable isotopes and fatty acids analysis) was applied in order to identify different fish species and trace their geographical provenience. The investigation was focused on four species (Thunnus thynnus, Thunnus alalunga, Auxis rochei and Scomber scombrus) belonging to the Scombridae family. The DNA barcoding method confirmed genus and species for S. scombrus and A. rochei, but only genus for T. alalunga and T. thynnus. Carbon and nitrogen stable isotopes results evidenced different fish diets and trophic positions, whereas fatty acids analysis displayed that the unsaturated prevailed (∼60 %) over the saturated compounds with a variation among the species and the geographical area in particular for docosahexaenoic and eicosapentaenoic acids percentage. The principal component analysis applied to stable isotopes and fatty acids evidenced a good discrimination among species and their geographical catching area. This multi-disciplinary analytical approach could represent a promising tool to identify the commercial fish and trace their origin in order to guarantee the health of consumers.

HPLC-UV/Vis-APCI-MS/MS Determination of Major Carotenoids and Their Bioaccessibility from "Delica" (Cucurbita maxima) and "Violina" (Cucurbita moschata) Pumpkins as Food Traceability Markers.

Carotenoids are a widespread group of fat-soluble pigments, and their major nutritional importance comes from their pro-vitamin A activity and their antioxidant capacity. In this study, two different pumpkin cultivars (Cucurbita maxima, also named `Delica' and Cucurbita moschata, also known as `Violina') from the southern Po Delta area were investigated in terms of carotenoid content and the influence of food processing on compositional changes and carotenoid bioaccessibility. Quali- and quantitative determination of carotenoids in sample extracts were performed on a C30 column by means of an online coupled HPLC-UV/Vis-APCI-MS/MS technique. The identification of separated compounds was tentatively achieved by merging (i) chromatographic data, (ii) UV-Vis spectra, and (iii) MS/MS fragmentation spectra. The chromatographic profiles for the two cultivars showed qualitative differences. Two major carotenoids were considered for quantification purposes and further investigations: lutein and ß -carotene. Quantification of target carotenoids was performed with external calibration through analytical standards. The concentration of lutein and ß -carotene was higher in C. maxima than in the other variety, C. moschata. Carotenoids are susceptible to degradation (isomerization and oxidation) during food processing (i.e., cooking), and the concentration of lutein and ß -carotene were monitored in oven-cooked and steam-cooked pumpkins. The steam-cooking process was superior in terms of limiting carotenoid loss. A complete functional profile of pumpkins as a source of carotenoids was gained with the evaluation of their in vitro bioaccessibility and their bioavailability after intake during human digestion. Bioaccessibility of lutein and ß -carotene were estimated by an in vitro static digestion model that involved salivary, gastric, and duodenal phases. Bioaccessibility values progressively increased from the salivary to the duodenal phase for both pumpkin varieties and cooking methods. Bioaccessibility of lutein was always lower than ß -carotene for both cultivars and for both cooking methods. Bioaccessibility values for lutein and ß -carotene changed from 1.93% to 2.34% vs. 4.94% and 8.83% in the salivary phase, from 2.7% to 4.63% vs. 7.83% and 15.60% in the gastric phase, and from 10.04% to 13.42% vs. 25.81% and 35.32% in the duodenal phase. For both target compounds, bioaccessibility in the duodenal phase was more than twice the gastric values, and it underlined that the type of cooking did not influence release from the initial matrix.

Metabolite profiling of flavonols and in vitro antioxidant activity of young shoots of wild Humulus lupulus L. (hop).

Humulus lupulus L., commonly named hop, is well-known for its sedative and estrogenic activity. While hop cones are widely characterized, only few works have been carried out on the young shoots of this plant. In the light of this, the aim of this study was to identify for the first time the flavonoids present in young hop shoots and to compare the composition of samples harvested from different locations in Northern Italy with their antioxidant activity. The samples were extracted by means of dynamic maceration with methanol. The HPLC-UV/DAD, HPLC-ESI-MS and MS2 analysis were carried out by using an Ascentis C18 column (250×4.6mm I.D., 5µm), with a mobile phase composed of 0.1M formic acid in both water and acetonitrile, under gradient elution. Quercetin and kaempferol glycosides were the main compounds identified and quantified in hop shoot extracts. Total flavonols ranged from 2698±185 to 517±48µg/g (fresh weight). The antioxidant activity was determined by means of the radical scavenging activity assay against diphenylpicrylhydrazyl (DPPH) and by using a photochemiluscence assay with a Photochem® apparatus. The results showed that hop shoots represent a new source of flavonols; therefore, they can be useful for a possible incorporation in the diet as a functional food or applied in the nutraceutical ambit.

Antioxidant capacity of trans-resveratrol dietary supplements alone or combined with the mycotoxin beauvericin.

Trans-resveratrol (trans-RSV) is a polyphenol with multiples biological properties, such as anti-inflammatory, antioxidant, anti-aging, anti-diabetic, and antiplatelet. It occurs naturally in grapes and derivate, peanuts and berries. Beauvericin (BEA) is a mycotoxin present in cereals that produces cytotoxicity, intracellular reactive oxygen species and lipid peroxidation. The general objective of this research was to evaluate whether trans-RSV could be used as a good polyphenol against damages produced by BEA. Because trans-RSV can be ingested through dietary supplements, to reach this goal, the following specific objectives were proposed: to determine a) the trans-RSV content in different polyphenol dietary supplements by capillary electrophoresis, b) the antioxidant capacity of the trans-RSV in polyphenol supplements, and c) the influence of BEA in the antioxidant capacity of trans-RSV when they are in combination by photochemioluminiscence assay. The results obtained in this study showed that all polyphenol dietary supplements present higher RSV content that the content of the label. The polyphenol supplements present antioxidant capacity. And the combination of trans-RSV and BEA did not affect the antioxidant capacity of trans-RSV. Thus, RSV could contribute to decrease oxidant effects produced by BEA.

Total Serum Antioxidant Capacity in Healthy Normal Weight and Asymptomatic Overweight Adults.

Obesity and overweight are major contributors to the burden of chronic disease. Both are defined as abnormal or excessive fat accumulation and by increased production of free radicals leading to oxidative stress. The aim of the present study was to evaluate whether overweight and fat accumulation is associated with serum total antioxidant capacity (TAC) in men and women, irrespective of nutritional habits, nutrient intakes, physical activity, smoking, and other confounders, which may be responsible for modifying the association between serum TAC and overweight/obesity measures. This cross-sectional study was conducted on 60 normal weight and 60 overweight adults aged 25-49. All participants underwent standard anthromorphological measurements of body composition, blood pressure and biochemical measurements, aerobic capabilities assessment and dietary intake evaluation. TAC was measured by using the photochemioluminescence method. All data were analysed with SPSS software. Men had higher values of TAC than women and concentrations of TAC were significantly higher in overweight subjects compared to normal weight subjects. In the present study TAC tended to be increased by various metabolic risk factors, especially overweight/obesity parameters (body mass index, body fat), inflammation and increased serum levels of Cysteine, irrespective of nutritional habits, nutrient intakes, physical activity and smoking. Overweight and obesity at an early stage may stimulate TAC. Therefore, the elevation of TAC in overweight adults may be a compensatory response to oxidative stress, generated by reactive oxygen species.

In vitro bioaccessibility, transepithelial transport and antioxidant activity of Urtica dioica L. phenolic compounds in nettle based food products.

Nettle (Urtica dioica L.) is a well-known plant with a wide historical background use of stems, roots and leaves. Nettle leaves are an excellent source of phenolic compounds, principally 3-caffeoylquinic acid (3-CQA), caffeoylmalic acid (CMA) and rutin. The aim of this work was to evaluate the bioaccessibility (BAC), the bioavailability (BAV) and the antioxidant activity of nettle phenolic compounds present in foods and supplements. The BAC of nettle phenolics was evaluated with an in vitro dynamic digestion of real food matrices: the type of food matrix and chemical characteristic affected the kinetics of release and solubilization, with the highest BAC after duodenal digestion. A study of duodenal trans epithelial transport evidenced low bioavailability of native forms of 3-CQA, CMA and rutin. Simulation of colonic metabolism confirmed that phenolic compounds are fermented by gut microflora, confirming the need for further investigations on the impact of phenolic compounds at the large intestine level. Photochemiluminescence assay of the simulated digestion fluids demonstrated that ingestion of Urtica based foods contributes to create an antioxidant environment against superoxide anion radicals in the entire gastrointestinal tract (GIT).

Re-cultivation of Neochloris oleoabundans in exhausted autotrophic and mixotrophic media: the potential role of polyamines and free fatty acids.

Neochloris oleoabundans (Chlorophyta) is widely considered one of the most promising microalgae for biotechnological applications. However, the large-scale production of microalgae requires large amounts of water. In this perspective, the possibility of using exhausted growth media for the re-cultivation of N. oleoabundans was investigated in order to simultaneously make the cultivation more economically feasible and environmentally sustainable. Experiments were performed by testing the following media: autotrophic exhausted medium (E+) and mixotrophic exhausted medium after cultivation with glucose (EG+) of N. oleoabundans cells grown in a 20-L photobioreactor (PBR). Both exhausted media were replenished with the same amounts of nitrate and phosphate as the control brackish medium (C). Growth kinetics, nitrate and phosphate consumption, photosynthetic pigments content, photosynthetic efficiency, cell morphology, and lipid production were evaluated. Moreover, the free fatty acid (FFA) composition of exhausted media and the polyamine (PA) concentrations of both algae and media were analyzed in order to test if some molecules, released into the medium, could influence algal growth and metabolism. Results showed that N. oleoabundans can efficiently grow in both exhausted media, if appropriately replenished with the main nutrients (E+ and EG+), especially in E+ and to the same extent as in C medium. Growth promotion of N. oleoabundans was attributed to PAs and alteration of the photosynthetic apparatus to FFAs. Taken together, results show that recycling growth medium is a suitable solution to obtain good N. oleoabundans biomass concentrations, while providing a more sustainable ecological impact on water resources.

Nematicidal activity of allylisothiocyanate from horseradish (Armoracia rusticana) roots against Meloidogyne incognita.

In recent years, there has been a great development in the search for new natural pesticides for crop protection aiming a partial or total replacement of currently used chemical nematicides. Glucosinolate breakdown products are volatile and are therefore good candidates for nematodes fumigants. In this article, the methanol-aqueous extract (1:1, w/v) of horseradish (Armoracia rusticana) fresh roots (MAH) was in vitro tested for nematicidal activity against second stage (J2) Meloidogyne incognita. The EC50 of MAH after 3 days of J2 immersion in test solutions was 251 ± 46 mg/L. The chemical composition analysis of the extract carried out by the GC-MS technique showed that allylisothicyanate was the most abundant compound. This pure compound induced J2 paralysis with an EC50 of 52.6 ± 45.6 and 6.6 ± 3.4 mg/L after 1 h and 3 days of incubation. The use of LC-MS/MS showed for the first time that horseradish root is rich in polyphenols. The study of isothiocyanate degradation in soil showed that allylisothiocyanate was the most quickly degradable compound (half-life <10 min), whereas no significant differences in half-life time were noted between degradation in regular and autoclaved soil.

Morphophysiological analyses of Neochloris oleoabundans (Chlorophyta) grown mixotrophically in a carbon-rich waste product.

Neochloris oleoabundans is considered one of the most promising oil-rich microalgae because of its ability to store lipids under nitrogen starvation. However, high biomass densities, required for applications on medium to large scale, are not reached in this condition of growth. As previous studies on other microalgae have shown that mixotrophy allows to obtain higher biomass in comparison to autotrophic cultures, we performed morphophysiological analyses in order to test the mixotrophic growth capability of N. oleoabundans. A carbon-rich manure derived from the apple vinegar production (AWP) was added to the medium. Cells were also cultivated under nutrient starvation (tap water), to observe the expected lipids accumulation, and combining AWP to water, to test the potential of this waste in a low-cost culture system. The results highlighted that AWP in the medium allowed to obtain the highest final cell density. Moreover, starch granules were stored inside chloroplast at the beginning of the experiment. The presence of AWP did not induce variations on light harvesting complex II (LHCII)-photosystem II (PSII) assembly, even if an interesting promotion of pigment synthesis in cells was observed. On the other hand, in starved cells, chloroplast degeneration, pigment content decrease, altered LHCII-PSII assembly and accumulation of high amount of lipid globules were observed, irrespective of the presence of AWP. The results suggest that mixotrophy promotes growth in N. oleoabundans and open up the possibility of using waste products from agri-food industries for this purpose. After growth, cells could be transferred under nutrient starvation to induce lipid accumulation.

Analytical traceability of melon (Cucumis melo var reticulatus): proximate composition, bioactive compounds, and antioxidant capacity in relation to cultivar, plant physiology state, and seasonal variability.

Two morphologically different cultivars of Italian melons (Baggio and Giusto) were characterized considering samples harvested in different times, at the beginning (BPP) and at the end of the physiological plant production period (EPP). Proximate composition, protein, minerals, pH, phenolic content, antioxidant capacity, ascorbic acid, carotenoids, condensed tannins, and flavonoids were measured, showing a significant decrease in EPP samples (phenolics, antioxidant capacity, condensed tannins, and flavonoids); ascorbic acid decreased in Giusto cv, carotenoids in Baggio cv. Mineral content increased in either the cultivars (EPP samples). Year-to-year difference was significantly highlighted; the plant growing cycle significantly affected the chemotype. Despite these effects, the Principal Component Analysis (PCA) permitted the discrimination of Baggio from Giusto cv, and the discrimination of BPP from EPP samples as well.

Flavones and structurally related 4-chromenones inhibit carbonic anhydrases by a different mechanism of action compared to coumarins.

An inhibition study of several carbonic anhydrase (CA, EC 4.2.1.1) isoforms with flavones and aminoflavones, compounds possessing a rather similar scaffold with the coumarins, recently discovered inhibitors of this enzyme, is reported. The natural product flavone and some of its hydroxylated derivatives did not show time-dependent inhibition of the CAs, sign that they are not hydrolyzed within the enzyme active site as the (thio)coumarins and lactones. These compounds were low micromolar inhibitors of hCA I, II, IX and XII, with K(I)s in the range of 1.88-9.07 µM. A series of substituted 2-amino-3-phenyl-4H-chromen-4-ones, incorporating chloro- and methoxy substituents in various positions of the heterocycle, were then prepared and assayed as hCA I and II inhibitors, showing activity in the micromolar range. Some of these derivatives, as well as cis+trans resveratrol, were then assayed for the inhibition of all catalytically active mammalian CA isoforms, hCA I, II, III, IV, VA, VB, VI, VII, IX, XII, XIII, XIV and mCA XV (h=human, m=murine enzyme). These derivatives inhibited these CAs in the submicromolar-low micromolar range. Flavones, although not as active as the coumarins, may be considered as interesting leads for the design of non-sulfonamide CA inhibitors.

Insecticidal activity and fungitoxicity of plant extracts and components of horseradish (Armoracia rusticana) and garlic (Allium sativum).

To avoid environmental pollution and health problems caused by the use of traditional synthetic pesticides, there is a trend to search for naturally occurring toxicants from plants. Among the compounds discussed for anti-fungal and insecticidal activity, the natural extracts from garlic and horseradish have attracted considerable attention. The objective of this study is to determine the insecticidal and anti-fungal activity of Armoracia rusticana and Allium sativum L. extracts against larvae of Aedes albopictus (Skuse) and some pathogenic fungi. For the insecticidal test, horseradish and garlic extracts were prepared from fresh plants (cultivated in Emilia Romagna region) in a solution of ethanol 80 % and the two different solutions were used at different concentrations (for the determination of the lethal dose) against the fourth instar mosquito's larvae. The fungicidal test was carried out by the agar plates technique using garlic and horseradish extracts in a 10 % ethanol solution against the following organisms: Sclerotium rolfsii Sacc., Trichoderma longibrachiatum, Botrytis cinerea Pers., Fusarium oxysporum Schlecht. and Fusarium culmorum (Wm. G. Sm.) Sacc. The first results demonstrated that the horseradish ethanol extracts present only a fungistatic activity against Sclerotium rolfsii Sacc., Fusarium oxysporum Schlecht. and F. culmorum (Wm.G. Sm) Sacc. while garlic extracts at the same concentration provided a good fungicidal activity above all against Botrytis cinerea Pers. and S. rolfsii. A. rusticana and A. sativum preparations showed also an interesting and significant insecticidal activity against larvae of A. albopictus, even if horseradish presented a higher efficacy (LC50 value of 2.34 g/L), approximately two times higher than garlic one (LC50 value of 4.48 g/L).