Relating Trait Variation to Species and Community Productivity in Contrasting Oro-Mediterranean Pastures: A 7-Years Study in the Pollino National Park (S-Italy).

Understanding how functional traits influence community assemblage and functioning is crucial for assessing the effects of global change on vegetation composition. We studied the functional composition (i.e., plant size (SIZE), leaf area (LA), specific leaf area (SLA), and leaf dry matter content (LDMC)) of a xerophile pasture and a mesophile grassland in southern Italy, and recorded species richness (SR), plant cover (COV) and flowering rates (FLOW) over a 7-year period. Both communities revealed the dominance of stress-tolerators, probably reflecting an adaptation to the Mediterranean climate. The functional classification of species distinguished three groups. Species from the mesophile community had larger SIZE and LA, while those from the xerophile pasture showed higher LDMC; SLA was not connected to the source community. Community-level analyses confirmed such patterns, but with higher SLA in the mesophile grassland. While SR was comparable, COV and FLOW varied between the communities. At the species level, LDMC was positively related to FLOW and the inter-annual variability of COV and FLOW. At the community level, SIZE, LA and SLA were positively related to COV, while LDMC was positively related to FLOW. Trait variations can significantly contribute to the xerophile-mesophile shift in Mediterranean mountain vegetation, by regulating the productivity of species and communities in the two contexts and, possibly, their responsiveness to global change.

Do marginal plant populations enhance the fitness of larger core units under ongoing climate change? Empirical insights from a rare carnation.

Assisted gene flow (AGF) can restore fitness in small plant populations. Due to climate change, current fitness patterns could vary in the future ecological scenario, as highly performant lineages can undergo maladaptation under the new climatic contexts. Peripheral populations have been argued to represent a potential source of species adaptation against climate change, but experimental evidence is poor. This paper considers the consequences of within- and between-population mating between a large core population and the southernmost population, the rare Dianthus guliae, to evaluate optimal AGF design under current and future conditions. We performed experimental self-pollinations and within- and between-population cross-pollinations to generate seed material and test its adaptive value to aridity. Seed germination, seedling growth and survival were measured under current and expected aridity. Effects of population type, pollination treatment and stress treatment on fitness components were analysed by generalized linear models. Relative measures of inbreeding depression and heterosis were taken under different stress treatments. Self-pollination reduced fitness for all the considered traits compared to within- and between-population cross-pollination. Under current aridity regime, the core population expressed higher fitness, and a larger magnitude of inbreeding depression. This indicated the core unit is close to its fitness optimum and could allow for restoring the fitness of the small peripheral population. Contrarily, under increased aridity, the fitness of outbred core lineages decreased, suggesting the rise of maladaptation. In this scenario, AGF from the small peripheral population enhanced the fitness of the core unit, whereas AGF from the core population promoted a fitness loss in the peripheral population. Hence, the small peripheral population could improve fitness of large core units versus climate change, while the contrary could be not true. Integrating reciprocal breeding programmes and fitness analyses under current and predicted ecological conditions can support optimal AGF design in a long-term perspective.

Salvia officinalis L. from Italy: A Comparative Chemical and Biological Study of Its Essential Oil in the Mediterranean Context.

Salvia officinalis L. (sage) is one of the most appreciated plants for its plethora of biologically active compounds. The objective of our research was a comparative study, in the Mediterranean context, of chemical composition, anticholinesterases, and antioxidant properties of essential oils (EOs) from sage collected in three areas (S1-S3) of Southern Italy. EOs were extracted by hydrodistillation and analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory properties were investigated by employing Ellman's method. Four in vitro assays, namely, 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), ferric-reducing ability power (FRAP), and ß-carotene bleaching tests, were used to study the antioxidant effects. Camphor (16.16-18.92%), 1,8-cineole (8.80-9.86%), ß-pinene (3.08-9.14%), camphene (6.27-8.08%), and α-thujone (1.17-9.26%) are identified as the most abundant constituents. However, the content of these constituents varied depending on environmental factors and pedoclimatic conditions. Principal component analysis (PCA) was performed. Based on Relative Antioxidant Capacity Index (RACI), S2 essential oil exhibited the highest radical potential with an IC50 value of 20.64 µg/mL in ABTS test and presented the highest protection of lipid peroxidation with IC50 values of 38.06 and 46.32 µg/mL after 30 and 60 min of incubation, respectively. The most promising inhibitory activity against BChE was found for S3 sample (IC50 of 33.13 µg/mL).

Corrigendum: Sicoli G, Passalacqua NG, De Giuseppe AB, Palermo AM, Pellegrino G (2019) A new species of Psathyrella (Psathyrellaceae, Agaricales) from Italy. MycoKeys 52: 89-102. https://doi.org/10.3897/mycokeys.52.31415.

[This corrects the article DOI: 10.3897/mycokeys.52.31415.].

A new species of Psathyrella (Psathyrellaceae, Agaricales) from Italy.

Sporophores of a new Psathyrella species have been reported for the first time as growing at the base of Cladiummariscus culms in the Botanical Garden of the University of Calabria, Rende, Cosenza, southern Italy. The fungus was initially identified as P.thujina (= P.almerensis) by means of both ecology and macro- and microscopic characteristics of the basidiomes, then referred to P.cladii-marisci sp. nov. after extraction, amplification, purification and analysis of the rDNA ITS region. We came to this conclusion after comparing our specimen with the descriptions of the taxa available in the literature for the genus Psathyrella.

Common medium versus advanced IVF medium for cryopreserved oocytes in heterologous cycles.

Granulocyte-macrophage colony-stimulation factor plays different crucial roles during embryo implantation and subsequent development. Here we aimed to evaluate the effects of embryo cell culture medium, with the inclusion of granulocyte-macrophage colony-stimulation factor (GM-CSF), on embryo development and pregnancy rate. To this end, we took advantage of our retrospective observational study to correlate the outcomes from two different culture media. We included in this study 25 unselected patient from our IVF Center that underwent heterologous IVF cycle with crypreserved oocytes. We analyze the fertilization rate, pregnancy rate, and embryo quality at different day of transfer obtained from two different media composition. Our results show that the rate of fertilization and the pregnancy rate were increased using medium added with this particular type of cytokines (GM-CSF).

Histo-anatomical leaf variations related to depth in Posidonia oceanica.

The purpose of our study is to explore the acclimation of Neptune seagrass (Posidonia oceanica (L.) Delile) to depth by characterising the histo-anatomical leaf modifications. P. oceanica is the dominant seagrass and main habitat constructor of seagrass meadows in the Mediterranean Sea. Meadows play an important biological and ecological role in marine ecosystems, serving as a habitat for a large diversity of species and an efficient erosion protection system for our coasts. Seagrasses are very sensitive to change in light availability and small changes can have significant effects on growth, abundance and distribution. In this study, we analyse changes in P. oceanica leaves collected at -5m, -15m and -25m depth in the Cirella meadow (Tyrrhenian coast, Southern Italy) in order to determine their depth-related histo-anatomical variation. Two main changes were observed at depth: (1) photosynthetic epidermal cells showed smaller chloroplasts but in the same number; and (2) leaves showed smaller epidermal cells and in greater number. Hence, the photosynthetic surface of P. oceanica leaves remains the same at different depths but pigment absorption efficiency can be significantly enhanced with depth. This response supports the differential photoacclimatory response of seagrasses with respect to terrestrial plants previously documented. Mesophyll cells are smaller with depth and more numerous, with a consequent increase in leaf density. The number of vascular bundles also increases, which allows improved functional efficiency of the transport system and solute exchange. Our study is a new contribution to the morpho-functional implications of the histo-anatomy of P. oceanica.

Berberis aetnensis and B. libanotica: a comparative study on the chemical composition, inhibitory effect on key enzymes linked to Alzheimer's disease and antioxidant activity.

OBJECTIVES: This study involves for the first time the evaluation of Berberis aetnensis C. Presl. and Berberis libanotica Ehrenb. ex C.K. Schneid. roots for anticholinesterase and antioxidant properties, with the aim to search compounds possibly useful for the treatment of Alzheimer's disease (AD). METHODS: Samples were evaluated for their ability to inhibit acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). The antioxidant activity was assayed by 2,2-diphenyl-1-picrylhydrazyl radical and ß-carotene bleaching tests. Berberine and palmatine were isolated by flash chromatography and identified by spectrometric methods as main constituents. Non-polar compounds were analysed by gas chromatography and gas chromatography-mass spectrometry. KEY FINDINGS: Extracts, fractions and isolated compounds inhibited AChE and BChE to varying degrees. The methanol fractions exhibited the strongest AChE inhibitory activity with inhibitory concentration 50% (IC50 ) of 7.6 and 16.9 µg/ml for B. aetnensis and B. libanotica, respectively. The alkaloid fraction of B. aetnensis inhibited AChE (IC50 of 24.5 µg/ml) and possessed the highest antioxidant activity (IC50 value of 2.2 µg/ml in ß-carotene-bleaching test after 30 min of incubation). Berberine was more potent of palmatine against AChE (IC50 of 2.2 and 7.4 µg/ml, respectively). CONCLUSIONS: These findings raise the possibility of developing B. aetnensis and B. libanotica as a promising candidate for the treatment of AD.

Antiproliferative activities on renal, prostate and melanoma cancer cell lines of Sarcopoterium spinosum aerial parts and its major constituent tormentic acid.

The search for improved cytotoxic agents continues to be an important line in the discovery of modern anticancer drugs. Sarcopoterium spinosum (L.) Spach is mentioned in ethnobotanical surveys as a medicinal plant used for the treatment of cancer. The aim of this study is to investigate and to compare the aerial parts of S. spinosum collected in Italy and Lebanon for their chemical composition and their antiproliferative activity against ACHN, C32, A375, MCF-7, LNCaP and HeLa human cancer cell lines using SRB assay. The main constituent tormentic acid was isolated by MPLC and characterized by spectroscopic techniques (NMR, MS). Non polar compounds were analyzed by GC and GC-MS. S. spinosum showed an interesting antiproliferative activity against ACHN and C32 cell lines with IC(50) values of 2.4 and 2.7 µg/ml for S. spinosum from Italy and Lebanon, respectively. Remarkable results were obtained also against A375 and LNCaP cell lines. The cytotoxicity against ACHN cell line could be partially attributed to tormentic acid that demonstrated a higher cytotoxicity than the positive control vinblastine. Close association between the radical scavenging activity (evaluated by DPPH and ABTS assay) and cytotoxicity was also demonstrated. This investigation demonstrated the potential cytotoxic activity of S. spinosum taking into account also that none of the tested extracts, fractions and isolated compound affected the proliferation of normal cell line 142BR. Tormentic acid, the major constituent isolated from S. spinosum, play an important role in the cytotoxicity exhibited by the extract.

Intricate variation patterns in the diploid-polyploid complex of Alyssum montanum-A. repens (Brassicaceae) in the Apennine Peninsula: evidence for long-term persistence and diversification.

PREMISE OF THE STUDY: The Apennine Peninsula, one of the three peninsulas of the European Mediterranean, is an important hotspot of genetic and species diversity, but studies devoted to plant evolution are still very scarce in this region. We studied the diploid-polyploid complex of Alyssum montanum-A. repens, focusing on Apennine and adjacent southwestern Alpine populations from southeastern France, with the aim of examining their taxonomic position and evolutionary patterns. METHODS: We employed AFLP markers and cpDNA sequences, along with cytotype determination using flow cytometry, and a multivariate morphometric approach. KEY RESULTS: The Italian and French populations formed two well-delimited groupings within the studied complex that were, in contrast to previous taxonomic treatments, clearly distinct from A. montanum. Populations from southeastern France represent A. orophilum, a previously described but abandoned species. Those from central and southern Italy correspond to A. diffusum, exhibiting high, geographically structured variation (central Apennines, Gargano, and southern Apennines/Calabria). This pattern coincides with hotspot refugial regions, in congruence with the "refugia-within-refugia" hypothesis, and is reflected here in the recognition of three subspecies within A. diffusum. CONCLUSIONS: We provide evidence for the presence of Mediterranean refugia for the studied Alyssum montanum-A. repens complex located in central and southern Italy, which, however, did not contribute to the postglacial colonization of Central Europe. Past extinctions, genetic bottlenecks, and recent expansion were inferred in Central Europe, while long-term accumulation of diversity as well as polyploidization occurred in the Apennines.

Acetylcholinesterase and butyrylcholinesterase inhibitory activity of Pinus species essential oils and their constituents.

This study aimed to investigate the acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activity of the essential oils from Pinus nigra subsp. nigra, P. nigra var. calabrica, and P. heldreichii subsp. leucodermis. This activity is relevant to the treatment of Alzheimer's disease (AD), since cholinesterase drugs are currently the only drugs available to treat AD. P. heldreichii subsp. leucodermis exhibited the most promising activity, with IC(50) values of 51.1 and 80.6 microg/mL against AChE and BChE, respectively. An interesting activity against AChE was also observed with P. nigra subsp. nigra essential oil, with an IC(50) value of 94.4 microg/mL. Essential oils were analyzed by GC and GC-MS with the purpose of investigating their relationships with the observed activities. Among the identified constituents, terpinolene, beta-phellandrene, linalyl acetate, trans-caryophyllene, and terpinen-4-ol were tested. trans-Caryophyllene and terpinen-4-ol inhibited BChE with IC(50) values of 78.6 and 107.6 microg/mL, respectively. beta-Phellandrene was selective against AChE (IC(50) value of 120.2 microg/mL).

In vitro cytotoxic effects of Senecio stabianus Lacaita (Asteraceae) on human cancer cell lines.

This study is aimed to evaluate the in vitro cytotoxicity of Senecio stabianus Lacaita (Asteraceae) against renal adenocarcinoma ACHN, hormone-dependent prostate carcinoma LNCaP, amelanotic melanoma C32 and human breast adenocarcinoma MCF-7 cell lines. The n-hexane extract showed an interesting activity with IC(50) values of 62.7 and 71.1 microg mL(-1) against C32 and LNCaP, respectively. Two compounds identified in the n-hexane extract, linalool and beta-caryophyllene, were found to be active against C32 cells (IC(50) values of 23.2 and 20.1 microg mL(-1), respectively). Among tested constituents, the highest activity was found when alpha-humulene was applied to LNCaP cells (IC(50) of 11.2 microg mL(-1)). In order to complete the phytochemical study on S. stabianus, which has not been investigated previously, the most characteristic secondary metabolites of Senecio genus, pyrrolizidine alkaloids (PAs), were analysed. Here we report the isolation, structural elucidation and identification of nine PAs.

Pyrrolizidine alkaloid profiles of the Senecio cineraria group (Asteraceae).

Alkaloid profiles of five Senecio species (Asteraceae), including S. ambiguus subsp. ambiguus, S. ambiguus subsp. nebrodensis, S. gibbosus subsp. bicolor, S. gibbosus subsp. gibbosus, and S. gibbosus subsp. cineraria, were studied. Eleven pyrrolizidine alkaloids were identified and their content was evaluated by GLC-MS and GLC analysis. Otosenine and florosenine were found to be the major alkaloids in all studied species. It is interesting that only S. ambiguus subsp. nebrodensis was characterized by a high content of the alkaloids jacobine, jacoline, jaconine, and jacozine.

Contribution to the knowledge of the veterinary science and of the ethnobotany in Calabria region (Southern Italy).

BACKGROUND: A series of preliminary research projects on plants used in Calabria (Southern Italy) in veterinary science and in other ethno-botanical fields (minor nourishment, domestic and handicraft sector) was carried out in the last twenty years. From the ethno-botanical point of view, Calabria is one of the most interesting region, since in the ancient times it was subject to the dominant cultures of several people (Greeks, Romans, Byzantines, Arabs, Normans etc.). Until some decades ago the road network was poorly developed and villages were isolated, so that the culture of the "subsistence" and some archaic customs were kept. METHODS: Data were collected by means of "open" interviews to farmers, shepherds and housewives in the last twenty years. More than 100 informants were interviewed, mostly over 50 years old. Plants were identified by local informants through gathering in the area or through examination of the fresh plants collected by the researchers. The collected data were compared with pharmacobotanical papers mainly of southern Italy and with other studies, in order to highlight novelties or concordances of uses. RESULTS: The use of 62 taxa distributed into 34 families are described. Among these, 8 are or were employed in veterinary science, 8 as anti-parasitic agents, 19 in minor nourishment, 5 as seasoning, 38 for other uses. Some toxic species for cattle are also mentioned. CONCLUSION: Among the major findings: the use of Helleborus bocconei for bronchitis of bovines and of Scrophularia canina for lameness in veterinary science; Nerium oleander and Urginea maritima as anti-parasitic agents; Epilobium angustifolium, Centaurea napifolia L. and C. sphaerocephala L. in minor nourishment.

Evidence for the existence of ganglioside molecules in the antigen of Entamoeba histolytica.

Gangliosides were found to be present in Entamoeba histolytica. They were extracted from lyophilized trophozoites of the pathogenic strain HM-1:IMSS and purified by high performance thin-layer chromatography. Two resorcinol-positive bands, comigrating with GM2 and GD1a were demonstrated, revealing the existence of ganglioside molecules in Entamoeba histolytica. The GM2 content, determined as lipid-bound sialic acid, was 1.5 micrograms/10(8) amoebae, the content of the GD1a comigrating band was 0.32 microgram/10(8) amoebae. The identity of the GM2 comigrating band was confirmed by TLC immunostaining, using the monoclonal anti-GM2 antibody GMB28. Furthermore, six out of ten anti-amoeba positive sera selectively reacted with the GM2 comigrating band, as revealed by immunostaining on TLC plates. Absorption tests revealed that preincubation of anti-amoeba positive sera with standard GM2 was followed by a significant decrease in the reaction with amoeba trophozoites by indirect immunofluorescence. These results demonstrate that a GM2 comigrating component of Entamoeba histolytica may be one of the antigens responsible for the appearance of circulating antibodies in patients with amoebiasis.

Anticardiolipin and anti-beta 2-GPI are two distinct populations of autoantibodies.

This study has been undertaken to assess whether anticardiolipin and anti-beta 2-GPI are two distinct populations of (auto)antibodies, and to clarify whether the beta 2-GPI region critical for phospholipid binding is also crucial for anti-beta 2-GPI reactivity. Fourteen of the 62 anticardiolipin (aCL) ELISA positive sera (22.6%) were positive for anti-beta 2-GPI by immunoblotting, 42 (67.7%) for aCL using TLC immunostaining. IgG fractions from 5 sera gave the same anticardiolipin reactivity detected by TLC immunostaining in the corresponding sera. All anti-beta 2-GPI-positive sera were reactive with the phenylthiocarbamyl derivative of the protein, indicating that binding of phenylisothiocyanate with lysine residues does not modify the molecule antigenicity. In addition, incubation of IgG fractions with the phospholipid binding site did not modify reactivity with beta 2-GPI. These findings demonstrate that: a) "true" antiphospholipid antibodies are detectable in patients' sera; b) aCL and anti-beta 2-GPI have a different immunological profile; c) the beta 2-GPI phospholipid-binding site is not the region recognized by the antibodies.

Cerebrospinal fluid antiganglioside antibodies in patients with AIDS.

In this study the presence of brain antiganglioside antibodies in the cerebrospinal fluid (CSF) of patients with HIV infection was analysed. CSF samples were collected from 45 patients with AIDS and from 45 anti-HIV negative subjects, 15 of whom presented aseptic meningitis. Nineteen AIDS patients had clinically well-documented encephalopathy. Thirteen of these patients had white matter lesions shown by magnetic resonance imaging (MRI). Both IgG and IgM antiganglioside antibodies were detected by immunostaining on thin layer chromatography plates in three CSF samples from AIDS patients with progressive encephalopathy with signs of a diffuse demyelination, as revealed by MRI. Two of these CSF samples reacted specifically with GM3, GM1 and GD1a and one with GD1a. In none of the HIV infected patients without demyelinating encephalopathy, but with opportunistic infections or cerebral lymphoma, nor in the anti-HIV negative control subjects were antiganglioside antibodies detected. No association with JCV DNA, CMV DNA, EBV DNA, detected by nested PCR, nor HIV antigen p24 was found. These findings show the presence of brain antiganglioside antibodies in the CSF of AIDS patients for the first time. However, the findings do not suggest relating the presence of these antibodies to HIV encephalopathy or particular viral agents, but indicate that the antibodies are detectable in subjects with progressive encephalopathy with a diffuse demyelination.