Diversity of Pleurotus spp. (Agaricomycetes) and Their Metabolites of Nutraceutical and Therapeutic Importance.

Pleurotus spp. have been gaining popularity as a source for the creation of functional foods, nutraceuticals and novel pharmaceuticals. Despite Pleurotus is a specious genus including 208 legitimate species, only a few of them such as P. ostreatus are commercially accessible. The genetic and metabolic diversity of Pleurotus both at specific and subspecific level is therefore of main concern for many researchers. In addition to the conventional morphological approach, molecular and biochemical markers have been greatly contributing to investigate these issues. In this study, samples from six Pleurotus species (P. eryngii is represented by three varieties) were molecularly identified and the phylogeny was inferred to assess the relationships between the various taxa. Strains in pure culture obtained from 6 out of 7 species were cultivated as mycelium in vitro to investigate the metabolites by untargeted LC-MS/MS-based metabolomics. The results pointed out species-specific metabolite patterns and highlighted a clear difference between the P. eryngii group and P. ostreatus, although the latter appears more versatile depending on the strain. This is the first study pointing out and comparing different metabolite patterns in Italian samples of Pleurotus species, including P. eryngii varieties.

Phytochemical and biological investigations on the pollen from industrial hemp male inflorescences.

Cannabis sativa L. belongs to the Cannabaceae family and includes annual, robust, fast-growing and generally dioecious plants. Industrial hemp, and particularly the inflorescences, has been recently recognized as important source of bioactive extracts with antioxidant and antimicrobial effects. The goal of the present study was to explore botanical, phytochemical, and biological properties of water and hydroalcoholic hemp pollen extracts from male inflorescences. The extracts from hemp pollen were found rich in phenolic compounds, such as hydroxytyrosol, coumaric acid, and hesperitin. The phenolic profile was also consistent with the observed scavenging/reducing, enzyme inhibitory, and antimicrobial properties of the extracts. Regarding the antimicrobial effects, Escherichia coli, Trycophyton rubrum, and T. tonsurans were the most sensitive to growth inhibitory effects (MIC values: 9.92-79.37 µg/mL) of the extracts. Whereas, null effects on prostate PC3 and myocyte C2C12 cell viability, in the range 1-1000 µg/mL, are consistent with MIC values and suggest extracts' biocompatibility. The experimental data obtained, which are not reflected in the literature as the topic of hemp pollen is almost completely unexplored, confirm the innovativeness of a product obtained directly from bees, which in the face of greater variability and complexity can reserve promising applications in food, pharmaceutical, and cosmetic sectors.

Antimicrobial, Antioxidant, and Antiproliferative Effects of Coronilla minima: An Unexplored Botanical Species.

Coronilla species, belonging to the Coronilla genus (Fabaceae), have long been used in traditional medicine for treating cold, diabetes, pain, and as cardiotonics. The goal of the present study was to explore the phytochemical composition and pharmaco-toxicological properties of C. minima. In this regard, phenolic content, scavenging/reducing properties and antimicrobial activity toward pathogen bacterial (Escherichia coli, Pseudomonas aeruginosa, Bacillus cereus, Staphylococcus aureus) and fungal strains (Candida albicans, C. tropicalis, Aspergillus tubigensis and A. minutus) were investigated. Extract effects on human colon cancer HCT116 cell viability were also assayed. Finally, a bioinformatics approach was conducted with the aim to identify putative microbial and human protein targets underlying antibacterial, antimycotic, and antiproliferative effects. Phytochemical investigation suggested that water extract is richer in terms of total flavonoid and phenol content, whereas the hydroalcoholic extract was revealed to be more potent as antioxidant agent. According to bioinformatics analysis, the antibacterial activity of the hydroalcoholic extract could be related to its content in resveratrol. The presence of resveratrol could also explain the hydroalcoholic extract efficacy in reducing HCT116 cell viability. In conclusion, the present study represents the first phytochemical and bio-pharmacological investigation about C. minima. Like other plants belonging to the Fabaceae family, C. minima revealed a good source of resveratrol, which could explain, albeit partially, the efficacy of the hydroalcoholic extract as antimicrobial, antioxidant, and antiproliferative agent.

Vepris macrophylla (Baker) I. Verd Essential Oil: An Antifungal Agent against Phytopathogenic Fungi.

Rutaceae are widely used in ethnomedicine to treat infectious diseases in humans and plants. In this study, the antifungal activity of the Vepris macrophylla leaf essential oil (VEO) and its main components, citral and citronellol, was evaluated against six phytopathogenic fungi. In addition, the possible action of VEO on the synthesis of mycotoxins was evaluated as well. To determine the antifungal activity of VEO we used the agar dilution method and VEO showed inhibitory activity against all the tested fungi. In particular, VEO resulted to be fungicidal against Phytophthora cryptogea and Fusarium avenaceum. For all other fungi VEO exhibited fungistatic activity and the weakest effect was observed on Alternaria solani. Citral was very effective against P. cryptogea, F. avenaceum, F. poae and F. graminearum. On the other hand, citronellol showed good activity towards P. cryptogea and F. avenaceum and weaker activity towards F. poae and F. graminearum. It can be concluded that VEO can be considered a promising antifungal agent, especially against P. cryptogea and F. avenaceum, suggesting a possible use in the formulation of new selective and natural fungicides.

Comprehensive approaches on the chemical constituents and pharmacological properties of flowers and leaves of American basil (Ocimum americanum L).

Ocimum americanum L. (Lamiaceae) is a common food condiment and also used in traditional medicine in the management of several human diseases. Nonetheless, there has been no effort to delineate the biological and phytochemical profiles of leaves and flowers prepared by different extractive solvents (ethyl acetate, methanol (MeOH), and water). The pharmacological potential of O. americanum extracts on pro-oxidant/pro-inflammatory mediators in rat colon specimens treated with lipopolysaccharide was investigated. In parallel, the inhibitory effects of the extracts on fungal and bacterial strains involved in ulcerative colitis were studied. Qualitative phytochemical analysis showed the presence of phenols, flavonoids, and tannins. Water extracts of flowers and leaves showed strong reducing and radicals scavenging potential. Both MeOH and ethyl acetate extracts of the leaves and flowers were able to inhibit acetylcholinesterase, butyrylcholinesterase, and tyrosinase. All the extracts inhibited the selected bacterial and fungal strains, while only ethyl acetate flower extract displayed antioxidant/anti-inflammatory effects in rat colon. The water and MeOH extracts stimulated colon lactate dehydrogenase (LDH) and serotonin (5-HT) and induced spontaneous migration of HCT116 cells. Future investigations should focus on the biological activity of isolated phytochemicals from the leaves and flowers of O. americanum, in order to clarify the mechanism(s) of action substantiating the observed pharmacological properties.

Chemical profiling and pharmaco-toxicological activity of Origanum sipyleum extracts: Exploring for novel sources for potential therapeutic agents.

The phytochemical, antiradical, and enzyme inhibition profile of three solvent extracts (ethyl acetate, methanol, water) of Origanum sipyleum were assessed. We also performed a pharmacological study in order to explore protective effects induced by extracts in inflamed colon. LC-MS analysis revealed that the extracts contained different classes of phenolics. The aqueous extract showed the highest antioxidant and acetylcholinesterase (AChE) inhibitory effects. Total phenol and flavonoid contents were highest in aqueous and ethyl acetate extract, respectively. All extracts were effective in reducing colon pro-oxidant and pro-inflammatory biomarkers. The extracts revealed also able to inhibit fungal and bacterial species involved in ulcerative colitis, including Candida albicans, Candida tropicalis, Staphylococcus aureus, and Staphylococcus thyphimurium. Finally, we also showed the antiproliferative effects exerted by the EA extracts on human colon cancer HCT116 cell line. Concluding, our results indicated that O. sipyleum extracts displayed promising therapeutic properties which warrants further validation. PRACTICAL APPLICATIONS: The present phytochemical and biological studies, including antioxidant, anti-inflammatory, and antimicrobic assessments, showed significant protective effects exerted by O. sipyleum extracts in an experimental model of ulcerative colitis. The results are intriguing and suggest potential applications O. sipyleum extracts as sources of natural agents for the management of clinical symptoms related to ulcerative colitis, characterized by increased burden of oxidative stress and microbiome dysbiosis.

Protective effects induced by alcoholic Phlomis fruticosa and Phlomis herba-venti extracts in isolated rat colon: Focus on antioxidant, anti-inflammatory, and antimicrobial activities in vitro.

Phlomis fruticosa L. and P. herba-venti are species belonging to the Lamiaceae family, which have been traditionally used to prepare tonic and digestive drinks. Multiple studies also demonstrated the inhibitory effects of P. fruticosa extracts and essential oil against oxidative/proinflammatory pathways and bacterial strains deeply involved in ulcerative colitis. Considering these findings, the present study evaluated the effects of alcoholic P. fruticosa and P. herba-venti leaf extracts in isolated rat colon challenged with Escherichia coli lipopolysaccharide (LPS), an ex vivo experimental paradigm of ulcerative colitis. In this context, we assayed colon levels of pro-oxidant and proinflammatory biomarkers, including nitrites, malondialdehyde (MDA), lactate dehydrogenase (LDH), and serotonin (5-HT). Additionally, the extracts have been tested in order to evaluate possible inhibitory effects on specific bacterial and fungal strains involved in ulcerative colitis. Alcoholic P. fruticosa and P. herba-venti extracts were able to blunt LPS-induced nitrite, MDA, 5-HT, and LDH levels in isolated rat colon. The same extracts also inhibited the growth of Pseudomonas aeruginosa, E. coli, Staphylococcus aureus, Candida albicans and Candida tropicalis. In conclusion, our findings show a potential role exerted by alcoholic P. fruticosa and P. herba-venti in managing the clinical symptoms related to ulcerative colitis.

Multiple pharmacognostic characterization on hemp commercial cultivars: Focus on inflorescence water extract activity.

One of the most promising economic perspectives of hemp production chain is female inflorescence valorization, despite there being actually no chemical composition or biological data from water fraction. In this context, the focus of this study is the evaluation of protective effects related to hemp water flower extracts from four commercial cultivars (Futura 75, Kc virtus, Carmagnola Cs and Villanova). We evaluated the phytochemical profile through validated spectrophotometric and HPLC methods. Then, we studied the biological activity on C2C12 and HCT116 cell lines, and in an ex vivo experimental model of ulcerative colitis, constituted by isolated LPS-stimulated colon. Particularly, we assayed the blunting effects induced by hemp water extract treatment on LPS-induced levels of nitrites, malondialdehyde (MDA), prostaglandin (PG)E2 and serotonin (5-HT). All tested cultivars displayed similar total phenolic and flavonoid profile. However, Futura 75 water extract displayed a better antioxidant and anti-inflammatory profile. Considering this, Futura 75 extract activity has been subsequently assayed on bacterial and fungal species involved in ulcerative colitis, finding a significant inhibition on C. albicans and selected Gram positive and negative bacterial strains. Concluding, our results support the potential efficacy of hemp inflorescence water extracts in managing the clinical symptoms related to ulcerative colitis.

Characterization of Biological Activities of Methanol Extract of Fuscoporia torulosa (Basidiomycetes) from Italy.

This work investigated the antiradical, antibacterial, and antifungal properties of a methanol extract from Fuscoporia torulosa (PeruMyc1834) fruit body from Central Italy. The ITS partial sequence of the F. torulosa strain matched at a 100% rate those of various strains belonging to the species. Two complementary test systems, namely α,α-diphenyl-ß-picrylhydrazyl (DPPH) radical scavenging and ß-carotene/linoleic acid systems, were used. Free radical scavenging, measured by DPPH assay on methanol extract, showed an activity of 38.17 ± 2.74% of Trolox activity. The methanol extract of F. torulosa inhibited the growth of Gram-positive bacteria, particularly those belonging to the genus Bacillus (i.e., B. subtilis and B. cereus), better than Gram-negative bacteria. All the fungal isolates used in the present study for antifungal activity inhibited their growth when a concentration of 2.27 mg mL-1 of methanol extract was used (minimum inhibitory concentration from < 0.28 to 2.27 mg mL-1). This study established that the extracted antioxidant and antimicrobial compounds produced by F. torulosa can be exploited in treating diseases that affect humans, plants, and animals. There is a need to carry out further study to establish the chemical composition of the methanol extract.

Overview of the Biological Activities of a Methanol Extract from Wild Red Belt Conk, Fomitopsis pinicola (Agaricomycetes), Fruiting Bodies from Central Italy.

Fomitopsis pinicola (Sw.) P. Karst. (Fomitopsidaceae) is a medicinal mushroom with a variety of healthy properties. In this study we tested the radical scavenging activity and antimicrobial and anticancer potential of methanol extracts of F. pinicola from central Italy. Molecular identification confirmed that the samples were F. pinicola; a Basic Local Alignment Search Tool search showed a close match (99% sequence identity) with European isolates of this species. The free radical scavenging capacities, measured by DPPH assay, showed that the extract activity was 3.5% that of Trolox. The MTT test, evaluated after 72 hours of treatment with increasing doses of extract (5-500 µg · mL-1), considerably inhibited proliferation in a dose-dependent manner in 2 human tumor cell lines. This reduction was coupled with a relevant induction of apoptosis in the human leukemia THP-1 cell line after 24 hours of treatment, but a relevant toxic effect occurred in the human colon adenocarcinoma HT29 cell line. The genotoxic potential of the methanol extracts was studied by single-cell gel electrophoresis of normal human leukocytes exposed to 20 µg extract at 37°C for 30 minutes; no DNA damage was observed. The F. pinicola methanol extract was found to have varying degrees of antifungal effects against the pathogenic fungi tested (minimum inhibitory concentration from 23.63 to 66.81 µg · mL-1). The results show that the tested F. pinicola extract has strong antimicrobial and chemo-preventive activities, but is a poor antioxidant.

Essential oil composition and biological activity from Artemisia caerulescens subsp. densiflora (Viv.) Gamisans ex Kerguélen & Lambinon (Asteraceae), an endemic species in the habitat of La Maddalena Archipelago.

The purpose of this study was to investigate the composition of the essential oil obtained from a population of Artemisia caerulescens subsp. densiflora growing in Razzoli, an island in the La Maddalena Archipelago (Sardinia, Italy). A. caerulescens sups. densiflora Viv. (Asteraceae), a wild herb, seldom studied in the Mediterranean, represents one of the many rare endemic species growing in North Sardinia. The essential oil composition was analysed by means of GC/MS analysis, which showed davana ethers as the major volatile components, accounting together for 17.5%, followed by (E)-nerolidol (4.5%), ß-oplopenone (3.3%), cis-sabinene hydrate (5.2%) and terpinen-4-ol (4.7%). The oil was tested for antioxidant activity by means of DPPH test, inhibition of lipid oxidation test and hypochlorous acid test, which showed a quite interesting scavenger capacity. For the first time, we reported the cytotoxic activity of the essential oil of A. caerulescens subsp. densiflora, against three human tumour cell lines (A375, MDA-MB231 and HCT116), with IC50 values in the range 5.20-7.61 µg/mL, which deserved further studies to support its use as chemopreventive agent. Finally, the antimicrobial activity of the essential oil, displayed on a panel of human pathogens, was very low.

Antioxidant and antiproliferative activity of Stachys glutinosa L. ethanol extract.

Ethanol extracts of Stachys glutinosa L. (Lamiaceae) were investigated for antioxidative properties, as well as antiproliferative action on various cell lines. The antioxidant activities were investigated by ABTS (2,2'-azinobis-3-ethylbenzothiazoline-6-sulphonic acid) assay, DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging, ß-carotene/linoleic acid assay, scavenging of hydrogen peroxide (horseradish peroxidase test), superoxide anion scavenging, and hypochlorous acid scavenging (taurine test). The antioxidant activity was reported as IC50 and reveals antioxidant effects. Antiproliferative effects were measured in vitro on three cell lines: HepG2 (human hepatocarcinoma), MCF7 (breast human adenocarcinoma) and C2C12 (mouse myoblast) cell lines by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The ethanol extract induced variations in cell viability on all cell lines tested. At 200 µg/mL, the effects on cell viability were - 23%, - 27% and - 37%, respectively, for C2C12, MCF7 and HepG2.

Antioxidant activity of methanol extract of Helichrysum foetidum Moench.

Methanol extract of Helichrysum foetidum Moench (Asteraceae) was investigated for antioxidative properties. The antioxidant activities were investigated by 2,2'-azinobis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assay, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging, ß-carotene/linoleic acid assay, scavenging of hydrogen peroxide (HRPO test), superoxide anion scavenging (S.A.S. test) and hypochlorous acid scavenging (taurine test). The antioxidant activity was reported as IC50 and reveals Trolox-like antioxidative effects.

Chemical composition and antibacterial activity of the essential oil from Mentha requienii Bentham.

The chemical composition of essential oil obtained by hydrodistillation of the fresh aerial parts of Mentha requienii Bentham (Lamiaceae) collected on the Gennargentu Mountains (Sardinia, Italy) has been investigated by gas chromatography and gas chromatography-mass spectrometry. The main constituents that resulted were pulegone (78%), menthone (0.5%), isomenthone (18%), isopulegone (1.3%) and limonene (1.76%). In vitro antifungal activity is evaluated in order to identify new means that could be helpful in the prevention of contamination in indoor environments.

Researching accessible and affordable treatment for common dermatological problems in developing countries. An Ethiopian experience.

BACKGROUND: Skin diseases are very common in rural and urban areas of Ethiopia, and traditional preparations of plant origin might represent the only alternative to synthetic drugs. Improving knowledge of traditional medicines and assessing their safety and effectiveness is necessary. METHODS: We conducted a two-arm, randomized, double-blind, placebo-controlled trial assessing the efficacy of some cosmetic herbal preparations for common dermatologic problems: a 3% thyme essential oil antifungal cream and a 10% chamomile extract cream for eczema-like lesions. RESULTS: Ten subjects (66.5%) treated with the 3% thyme active cream were completely healed vs. four subjects (28.5%) from the placebo group (P=0.040). A large number of subjects treated with the chamomile cream were healed or improved, but no significant differences were found between active cream and placebo. A high rate of treatment acceptance was registered in both groups, no adverse effects were reported. CONCLUSIONS: A 3% thyme essential oil cream could represent a cheap and easily available opportunity to treat and heal mild to moderate cases of fungal infections; a common emollient cream could be a very effective intervention when treating mild to moderate cases of pityriasis alba and eczema-like lesions. Further research is needed.

Chemical composition and inhibitory activity against Helicobacter pylori of the essential oil of Apium nodiflorum (Apiaceae).

The chemical composition of the essential oil obtained from Apium nodiflorum (L.) Lag. (Family Apiaceae), a plant used in the ethnomedical traditions of the Abruzzo region (Central Italy) as a culinary herb, as a diuretic, and to cure stomachache, was analyzed by gas chromatography/mass spectrometry, and 14 components were identified. Limonene (27.72 %), p-cymene (23.06%), myristicine (18.51%), and beta-pinene (6.62%) were the main components. The antimicrobial activity of the essential oil was assayed in vitro against Helicobacter pylori (strain DSMZ 4867), resulting in a minimum inhibitory concentration value of 12.5 microg/mL.

Chemical composition and antimicrobial activity of the essential oils from several Hypericum taxa (Guttiferae) growing in central Italy (Appennino Umbro-Marchigiano).

The chemical composition of the essential oils of nine taxa from seven sections of Hypericum L. (Guttiferae; H. perforatum subsp. perforatum, H. perforatum subsp. veronense, H. calycinum, H. montanum, H. richeri subsp. richeri, H. hyssopifolium, H. hirsutum, H. hircinum subsp. majus, and H. tetrapterum) occurring in central Italy (Appennino Umbro-Marchigiano) was analyzed by GC/FID and GC/MS. A total of 186 compounds were identified in the different species and subspecies, accounting for 86.9-92.8% of the total oils. The major fraction of the oil was always represented by sesquiterpene hydrocarbons (30.3-77.2%), while quantitative differences occurred between the other classes of volatiles depending on the taxa considered. Chemical composition of the nine Hypericum entities with respect to the taxonomical classification was discussed. Essential oils obtained from six taxa, i.e., H. perforatum subsp. perforatum, H. perforatum subsp. veronense, H. calycinum, H. richeri subsp. richeri, H. hirsutum and H. tetrapterum, were also tested for their antimicrobial properties against five different microbial strains by the broth-microdilution method, and they were found to have significant activity (expressed as MIC) on B. subtilis, moderate activity on C. albicans and S. aureus, and weak activity on E. coli and E. faecalis, the most active being those from H. hirsutum, H. richeri subsp. richeri, and H. tetrapterum.

The role of secreting structures position on the leaf volatile organic compounds of Hypericum androsaemum.

Hypericum androsaemum L. presents typical translucent, essential oil producing glands, which are distributed on the leaf along both margins (margin glands) and on the lamina (lamina glands). The gland secretion was studied by histochemical and chemical analysis; the gland content was sampled directly from the secretory glands, and the volatile organic compounds (VOC) of the margin and lamina glands were separately analyzed. The lipophilic fraction of the lamina glands had as main components: (E)-2-hexenal (15.5%), hexadecanoic acid (14.7%), beta-caryophyllene (11.2%), germacrene B (11.0%) and gamma-himachalene (9.8%). The lipophilic fraction of the margin glands had as its main components: beta-pinene (22.0%), limonene (17.6%), (E)-beta-ocimene (6.1%), methyl linoleate (5.7%), terpinolene (5.4 %), (E)-2-hexenal (4.9%) and alpha-pinene (4.1%).

Composition of essential oils from leaves and flowers of Stachys germanica subsp. salviifolia (Ten.) gams (Labiatae) and related secretory structures.

The essential oils from both leaves (L) and flowers (F) of Stachys germanica subsp. salviifolia (Ten.) Gams (Labiatae/Lamiaceae) from Italy were analyzed by GC-MS for the first time. The distribution, morphology and histochemistry of the different types of glandular trichomes present on the epidermal surface were also determined. Twenty-nine constituents, representing 89.4% of the total leaf oil, and forty-one compounds, forming 97.8% of the total flower oil, were identified. Germacrene D (39.4%), phytol (10.2%), beta-bourbonene (3.5%) and beta-ylangene (3.3%) were recognized as the main constituents of the leaf essential oil, while limonene (24.1%), beta-pinene (18.7%), germacrene D (12.8%) and (E)-nerolidol (6.6%) were the main compounds of the flower essential oil.

Rosmarinus officinalis L.: chemical modifications of the essential oil and evaluation of antioxidant and antimicrobial activity.

Rosmarinus officinalis essential oil was separated into its hydrocarbon and oxygenated fractions. The major compounds in the hydrocarbon fraction were alpha-pinene (44.2%), camphene (24.5%), and limonene (11.7%), while in the oxygenated fraction they were 1,8-cineole (37.6%), camphor (16.5%), and bornyl acetate (21.4%). The hydrocarbon fraction was submitted to a hydroformylation process and the antioxidant activity of the product was screened by the DPPH and beta-carotene/linoleic acid tests. The hydroformylated fraction maintained the antioxidant activity of the whole oil. The MIC (minimal inhibitory concentration) and the MBC (minimal bactericidal concentration) of the essential oil, hydrocarbon, oxygenated and hydroformylated fractions were also tested on several microorganisms. Aeromonas sobria and Candida strains were the most susceptible micro-organisms. The hydroformylated fraction exhibited a MBC against Candida strains resistant to the other fractions.