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The importance of taking into account ecosystems, plant communities and habitats for the development of biodiversity conservation strategies is increasingly acknowledged. Recently, the first ever European Red List of Habitats was produced, which provided an evaluation of the extinction risk of EUNIS-based natural and semi-natural habitats in Europe. As assessment unit, it used the habitat intended as a plant community, thus representing a landmark for the role of vegetation science in nature conservation. In the present paper, the results of the European Red List of Habitats are analyzed at the national scale with specific reference to the terrestrial and freshwater habitat types occurring in Italy. More than three-quarters of the assessed European habitat types were recognized for the Italian territory. The distribution of the threat categories reflects approximately the situation at the EU28 level. About 35% of the assessed habitat types are referred to a threat category; no critically endangered habitat is present in Italy. The most frequently used criteria are those related to a reduction in quantity. Some critical issues arising from the analyses are discussed. In particular, the presence of knowledge gaps is pointed out, with remarkable reference to the poor availability of spatial and quantitative data, severely affecting the application of the criteria adopted for the assessment. Descriptions of habitat types from Italy are reported, some of which are representative, emblematic or even exclusive to the Italian territory. The outcomes of the analysis represent the starting point for the future development of a national-scale Red List of Habitats. Results also emphasized how habitat types with a too broad definition pose a limit to a proper evaluation of the regional biogeographic variability, often very high in Italy, with local floristic and phytocoenotic peculiarities which do not find room in the adopted European typology. This is the reason why the development of national subtypes stands as a necessary step for the development of a realistic and effective assessment at the national scale.

Human papilloma virus (HPV) infection is the most prevalent among sexually transmitted diseases (STDs). There are more than 100 types of HPV, with the possibility of infection for both male and female, affecting sexual organs, mouth, and throat. The majority of people affected are unaware of the infection. HPV is associated with a large spectrum of clinical features, such as unapparent infections, unspecific epithelial lesions, genital warts, and cancers. Although a consistent and coherent picture about the epidemiology and pathogenesis of genital HPV infections in women has been developed over the past 30 years, fewer studies have been conducted on this infection in men. The same has happened with regards to the vaccine, which were concentrated on females with the primary objective being the prevention of cervical cancer. In recent years, with a better understanding of HPV infection and its clinical manifestations in males, the quadrivalent vaccine has been approved signifying a more forward-thinking approach in this population. More targeted interventions are needed for the control of HPV infection in men, particularly those who are unaware and probably account for most transmissions. The purpose of this review is to give an insight on the most recent evidence available in the literature on HPV treatment and prevention among men.

A 23-year-old woman was referred to our center with hirsutism, acne, weight gain, weakness, and irregular menses. Laboratory tests revealed increased levels of cortisol and sex hormones, and reduced adrenocorticotropic hormone levels. The patient underwent a right adrenalectomy. Pathology of the resected right adrenal gland showed an adrenal carcinoma. Computed tomography 8 months after the surgery revealed a thymic mass that was not detected preoperatively. The frequency of rebound thymic hyperplasia after normalization of hypercortisolism is not known, but possibly as high as 40%. Physicians must be aware of this phenomenon to avoid unnecessary thoracic surgery.

AbstractThere is an increasing interest in the taxonomy and distribution of the forms of the Portulaca oleraceacomplex. The information accruing from specimens collected in the Italian peninsula and surrounding islands (Sicily, Sardinia, Corsica and Malta) is here described. Eleven morphotypes were recorded: ‘P. cypria’, ‘P. granulatostellulata’, ‘P. nitida’, ‘P. oleracea’, ‘P. papillatostellulata’, ‘P. rausii’, ‘P. sardoa’, ‘P. sativa’, ‘P. sicula’, ‘P. trituberculata’, ‘P. zaffranii’ and a still unclear form Portulaca oleraceaf. Three occur in almost all the Italian peninsula and adjacent islands; three are scattered in the Italian peninsula and in the adjacent islands; the remnant have a distribution restricted to the islands such as Sicily and Sardinia. The morphotypes can be divided into two main categories: seeds smooth, without ornamentations; seeds with ornamentations. The morphotypes with ornamentations are more widespread than smooth ones, probably because seed ornamentations play an important role in seed dispersal, which is perhaps mainly anthropochorous given that P. oleraceais a synanthropic species that can tolerate mechanical disturbance. There are cases of multiple occurrence, as evidenced by the presence of different morphotypes in some of the sites. Such populations can count up to five morphotypes growing together. Seven morphotypes were here recorded from Malta; they are all hexaploid, even those which in other areas are tetraploid.

Background: There is a long-term trend towards the abandonment of agro-pastoral activities in the mountain areas of Europe: the following encroachment process of semi-natural grasslands by shrubs is one of the main severe threats to the conservation of biodiversity in mountain environments. To better understand the impact of land abandonment, we analysed the reliability of plant functional groups, ant traits, and ant functional groups as indicators of land use changes. We carried out the research in Italy at four sites along a latitudinal/altitudinal gradient in three biogeographic regions (Mediterranean, Continental, Alpine). We identified three stages of a chronosequence at each site as representative of the plant succession in response to pastoral land-use abandonment. Results: As expected, both the plant and ant assemblages considerably differed across sites at the species level and, within each site, among the three stages. This trend was found also using ant traits, functional groups of ants, and plant functional groups. Ant and plant communities were related in terms of composition and functionality. Harvester ants and ants with collective foraging strategy were associated with annual legumes and grasses (Therophytes); ants with a strictly individual foraging strategy with Phanerophytes. Ant traits and plant functional groups indicated significant differences among the three stages of the chronosequence. However, ant functional groups could not discriminate between the stages represented by secondary grasslands currently grazed and shrub-encroached grasslands ungrazed. Conclusion: Despite some limitations of ant functional groups in explaining the succession stages of land abandonment, our results suggest that ants are a good surrogate taxon and might be used as bioindicators of land-use changes and ecosystem functioning. Furthermore, our findings indicate that the functional group approach should be applied to other European ecosystems. Finally, reducing the taxonomic complexity could contribute to developing predictive models to detect early environmental changes and biodiversity loss in mountain habitats.

We performed a real-life observation of patients with Parkinson disease (PD) who received duodenal levodopa infusion (DLI) to determine which adverse events caused treatment discontinuation and when such events occurred.

Polycomb group proteins represent a global silencing system involved in embryonic development and stem-cell maintenance that regulates the transition from proliferation to differentiation during organogenesis. Two main complexes have been discovered: the polycomb repressive complex (PRC) 1 and 2, able to induce gene silencing by a synergistic mechanism or independently by each other. Enhancer of zeste homolog 2 (EZH2), the catalytic subunit of PRC2, represses gene transcription through the tri-methylation of histone H3 lysine 27. EZH2 deregulation is frequently associated with tumorigenesis, metastatic character, and poor prognosis in various cancer types. This review explores the role of EZH2 in normal development and in carcinogenesis. We reviewed the polycomb-mediated silencing mechanisms, the regulation of EZH2 activity and its recruitment to target genes. We also analyzed the role of EZH2 in normal muscle differentiation and in rhabdomyosarcoma, considering EZH2 blockade as a new strategy for developing specific therapies.

Rhabdomyosarcoma (RMS) is a pediatric tumor that arises from muscle precursor cells. RMS cells express several markers of early myogenic differentiation, but they fail to complete both differentiation program and cell cycle arrest, resulting in uncontrolled proliferation and incomplete myogenesis. Previous studies showed that EZH2, which is involved in both differentiation and cancer progression, is overexpressed in RMS, but a functional binding between its expression and its functional role in tumor formation or progression has not yet been demonstrated. We hypothesized that EZH2 is a key regulator of muscular differentiation program in RMS cells.In this study, we demonstrated that EZH2 directly binds muscle specific genes in RD cells. Silencing of EZH2 promotes the recruitment of a multiprotein complex at muscle-specific promoters, their transcriptional activation and protein expression. Moreover, we demonstrated that EZH2 is directly involved in transcriptional repression of MyoD, the main factor promoting myogenesis. EZH2 ablation induces MyoD activation the recovery of its binding on muscle-specific genes.

The aim of this study was to evaluate and compare the dynamics of plant, crustacean, and coleopteran assemblages living in Mediterranean temporary wetlands. We measured assemblage parameters (species richness, diversity and abundance) and succession rates during the hydroperiod for each assemblage analysed. Moreover, the species were classified according to their affinity to the temporary aquatic habitats in order to study if each assemblage had similar patterns among species groups of habitat affinity. Only among plants, the temporary water species were dominant in terms of species richness, diversity or abundance. Cross-taxon congruence between plants and crustaceans, previously reported, was not confirmed when temporal succession was taken into account, suggesting different temporal patterns for the studied taxa. Plants showed the lowest level of successional changes and significant differences between local and regional diversity. In contrast, coleopterans had the highest level of successional changes and a different relationship between local and regional diversity, with regard to the hydroperiod phase. Finally, crustaceans had intermediate levels of successional changes and the relationship between the local and regional diversity during the hydroperiod was dependent on the type of organism (generalist vs. temporary water species). All of this gave evidence of the difficulties to use one taxonomic group in order to elucidate the ecological patterns of other groups.

The aim of this study was to evaluate and compare the dynamics of plant, crustacean, and coleopteran assemblages living in Mediterranean temporary wetlands. We measured assemblage parameters (species richness, diversity and abundance) and succession rates during the hydroperiod for each assemblage analysed. Moreover, the species were classified according to their affinity to the temporary aquatic habitats in order to study if each assemblage had similar patterns among species groups of habitat affinity. Only among plants, the temporary water species were dominant in terms of species richness, diversity or abundance. Cross-taxon congruence between plants and crustaceans, previously reported, was not confirmed when temporal succession was taken into account, suggesting different temporal patterns for the studied taxa. Plants showed the lowest level of successional changes and significant differences between local and regional diversity. In contrast, coleopterans had the highest level of successional changes and a different relationship between local and regional diversity, with regard to the hydroperiod phase. Finally, crustaceans had intermediate levels of successional changes and the relationship between the local and regional diversity during the hydroperiod was dependent on the type of organism (generalist vs. temporary water species). All of this gave evidence of the difficulties to use one taxonomic group in order to elucidate the ecological patterns of other groups.

SummaryCalcareous outcrops in North-Western Sardinia cover a large area (about 380 Km) known as “Sassarese”. in this study the vascular flora of this area is investigated. The actual flora is checked and, when possible, older references, synonyms and critical notes are given. The floristic list includes 840 entities. Among them, 18 have been cultivated (cultae), 11 are considered erroneously reported and so they are excluded from this area (excludendae) and 81 are in various ways considered not confirmed and to enquiry (inquirendae).The current floristic contingent is therefore reduced to 730 entities, divided into 98 families and 385 genera, many of them here documented for the first time in this area. Among them the more relevant are the Sardinian-Corsican endemic Silene nodulosaViv. and the Sardinian endemic Hieracium gallurenseArrigoni, previously reported only for granite substrata in North-Eastern Sardinia. The biological spectrum shows a clear dominance of therophytes (41%). The chorological spectrum highlights the dominance of the Tethydic element, in which the dominance of more strictly Mediterranean element emerges. Four entities have in this area their locus classicus: Centaurea corensisValsecchi et Filigheddu, Limonium racemosum(Lojac.) Diana and Scrophularia morisiiValsecchi, exclusive of the region object of this study, and Ophrys sphegodesMill. subsp. praecoxCorrias. Some entities, though not endemic, have a limited or fragmentary distribution in Sardinia, e.g. Capparis spinosaL. subsp. rupestris(Sm.) Nyman, Coridothymus capi- tatus(L.) Reichenb. f. and Erica multifloraL., whereas Viola arborescensL. Borago pygmaea(DC.) Chater et Greuter and Carex panormitanaGuss. are included in the Red Book of Italian Plants. Although the area is strongly urbanised and subjected since long time ago to agricultural and pastoral uses, its flora is still rather diversified. The presence of species depending on the existence of hedges, dry-stone walls and other smaller structures belonging to traditional agricultural activities still present, is also substantial, even though their dilapidated state is often high. Finally the presence of floristic elements of great interest together with small residuals of spontaneous vegetation are of very high value. indeed, several natural elements from traditionally agricultural activities are present within an area of intense anthropic impact. The indiscriminate building development of the valleys, the reckless exploitation of the water sources and the diffusion of dumps, threaten increasingly some habitats of the region. The improvement of the natural resources could be done through an adequate ecological net starting from a basic knowledge of the floristic biodiversity, that is the main aim of this work.

Recent theoretical analyses demonstrate how informational asymmetries between financiers and investors may generate credit rationing and positive cost differentials between external and internal financing sources. The traditional empirical approach used to test for the presence of financing constraints at firm level is based on two pillars: a prioriidentification of relatively more financially constrained firms and econometric estimation of an investment demand function. This approach has been seriously questioned due to several methodological problems. This paper intends to amend it by adding a third pillar: the informational content of direct revelation through qualitative data. The paper estimates a reduced form investment equation following the Euler equation approach, and combines a prioriinformation and direct qualitative information to consistently estimate for each firm the probability of being financially constrained. Our main finding is that when financially constrained firms are properly identified, the neoclassical model is rejected only for unconstrained firms. This indirectly rescues the validity of the Euler equation approach. Moreover, financially constrained firms show a positive correlation between investment and lagged cash flow.

Objectives: The cyclin-dependent kinase p16 (also known as Ink4A, Mts1, Cdkn2, and Cdkn4i) has been proposed as a tumor suppressor gene mapped on chromosome segment 9p21. This study evaluated p16 protein expression in 135 lung cancer specimens and investigated potential genetic alterations occurring in this gene. Results: We found altered p16 immunohistochemical expression to be a frequent event in lung cancer and to be independent of either the histologic type or any other clinical-pathologic feature. Western blot analyses performed on about one third of the specimens correlated highly with these results. In addition, we found p16 immunohistochemical expression to be a favorable prognostic factor in lung cancer in that its reduction or loss correlated with a worse outcome for the patients. Polymerase chain reaction amplification and direct sequencing of p16 exons 1 and 2 revealed no mutations, indicating that p16-altered expression in lung cancer is not necessarily linked to mutational events of these genes. Conclusions: We conclude that p16-altered expression is both an independent and frequent event in lung cancer and may have an important role in tumorigenesis and in malignant progression of a significant proportion of these cancers. However, the actual incidence and relevance of p16 mutations in this neoplasm continues to be debated, and its analysis seems inconclusive. Our results suggest a prognostic role for the immunodetection of this protein on formalin-fixed and paraffin-embedded specimens. They further suggest its routine use in the evaluation of the frequently unpredictable behavior of lung cancer. (J Thorac Cardiovasc Surg 1999; 118:529-35)

Cathelicidins are a family of myeloid antimicrobial peptide precursors that have been identified in several mammalian species (Zanetti, M., Gennaro, R., and Romeo, D. (1995) FEBS Lett. 374, 1-5). Two novel bovine congeners have been deduced from cDNA. Their C-terminal sequences of 27 and 28 residues correspond to putative antimicrobial peptides with a cationic N-terminal region predicted to assume an amphipathic α-helical conformation followed by a hydrophobic C-terminal tail. Peptides corresponding to these sequences have been chemically synthesized and shown to exert a potent antimicrobial activity against Gram-negative and Gram-positive bacteria, including methicillin-resistant Staphylococcus aureus, and fungi. Both peptides are also cytotoxic to human erythrocytes and neutrophils, although at higher than microbicidal concentrations. The target selectivity has been improved by synthesizing truncated analogues, comprising only the 18 N-terminal residues, which show a great reduction in cytotoxic, but not in antimicrobial activity. The involvement of the C-terminal hydrophobic tail in the cytotoxic activity has been further demonstrated by inducing a major loss of activity in an analogue after replacing highly hydrophobic residues with more hydrophilic ones.

Several myeloid antimicrobial peptide precursors have been shown to consist of a N-terminal proregion similar to a protein named cathelin and a structurally varied C-terminal antimicrobial domain. Proteins with these features have been named cathelicidins. In this paper we report the cDNA sequences of three ovine cathelicidins of 155, 160 and 190 residues, respectively, with cationic C-terminal sequences corresponding to putative antimicrobial domains. These are structurally varied and include a Cys-rich sequence of 12 residues, which is similar to the bovine antimicrobial cyclic dodecapeptide, a novel 29 residue sequence named SMAP-29 with a possible α-helical conformation, and a 60 residue sequence named Bac7.5, which appears to be a new member of the Pro- and Arg-rich group of mammalian antimicrobial peptides.

Reduced state support of film financing in Europe requires the creation of an efficient market of private cinema financiers. Principal-agent models with and without common consent over film risk show that the two main problems of film financing (high risk and lack of collateral for the borrower) may be solved if the financier achieves economies of scale in costs of capital and is efficient in information gathering. Conversely, the pathology of a market with “inefficient financiers” includes: film financial rationing, adverse selection, and refusal of financial support to otherwise profitable film projects. From the normative point of view, the creation of efficient financiers would be fostered by institutional changes that promote market thickening and “risk spreading” (the case of the USA) instead of simple forms of “risk shifting” (the case of Italy).

The Cdk2 kinase has long been known to be involved in the progression of mammalian cells past the G1 phase restriction point and through DNA replication in the cell cycle. The Rb family of proteins, consisting of pRb, p107, and pRb2/p130, has also been shown to monitor progression of G1 phase, mostly through their interaction with E2F family members. p107 is able to inhibit Cdk2 kinase activity through this interaction via a p21-related domain present in the C terminus of the protein. We show here that pRb2/p130 also possesses this activity, but through a separate domain. Moreover, we correlate the increased expression of pRb2/p130 during various cellular processes with the decreased kinase activity of Cdk2. We hypothesize that pRb2/p130 may act not only to bind and modify E2F activity, but also to inhibit Cdk2 kinase activity in concert with p21 in a manner different from p107.

The oncoprotein of simian virus-40, SV40 large T-antigen (Tag), is reported to target and to inactivate growth suppressive proteins such as the retinoblastoma family1–3and p53 (ref. 4, 5), leading to transformation of human cell lines in vitro, tumor production in rodents6, and detection of Tag in several human cancers including mesotheliomas7,8. The retinoblastoma family contains three members, pRb, p107 and pRb2/p130 (ref. 9), that are phosphorylated in a cell cycle-dependent manner10,11, have cell growth suppressive properties12and bind to specific members of the E2F family and various cyclins13. Even though mesotheliomas are among the most aggressive human cancers, alterations of important cell-cycle “controllers,” such as the Rb family genes, have never been reported in these tumors. We found the presence of SV40-like sequences in 86% of 35 archival specimens of mesothelioma. We also demonstrated that SV40 Tag, isolated from frozen biopsies of human mesothelioma, binds each of the retinoblastoma family proteins, pRb, p107 and pRb2/p130, in four of four specimens. We propose that the tumorigenic potential of SV40 Tag in some human mesotheliomas may arise from its ability to interact with and thereby inactivate several tumor and/or growth suppressive proteins.

The Cdk2 kinase has long been known to be involved in the progression of mammalian cells past the G1phase restriction point and through DNA replication in the cell cycle. The Rb family of proteins, consisting of pRb, p107, and pRb2/p130, has also been shown to monitor progression of G1phase, mostly through their interaction with E2F family members. p107 is able to inhibit Cdk2 kinase activity through this interaction via a p21-related domain present in the C terminus of the protein. We show here that pRb2/p130 also possesses this activity, but through a separate domain. Moreover, we correlate the increased expression of pRb2/p130 during various cellular processes with the decreased kinase activity of Cdk2. We hypothesize that pRb2/p130 may act not only to bind and modify E2F activity, but also to inhibit Cdk2 kinase activity in concert with p21 in a manner different from p107.

Cathelicidins are a family of myeloid antimicrobial peptide precursors that have been identified in several mammalian species (Zanetti, M., Gennaro, R., and Romeo, D. (1995) FEBS Lett. 374, 1-5). Two novel bovine congeners have been deduced from cDNA. Their C-terminal sequences of 27 and 28 residues correspond to putative antimicrobial peptides with a cationic N-terminal region predicted to assume an amphipathic alpha-helical conformation followed by a hydrophobic C-terminal tail. Peptides corresponding to these sequences have been chemically synthesized and shown to exert a potent antimicrobial activity against Gram-negative and Gram-positive bacteria, including methicillin-resistant Staphylococcus aureus, and fungi. Both peptides are also cytotoxic to human erythrocytes and neutrophils, although at higher than microbicidal concentrations. The target selectivity has been improved by synthesizing truncated analogues, comprising only the 18 N-terminal residues, which show a great reduction in cytotoxic, but not in antimicrobial activity. The involvement of the C-terminal hydrophobic tail in the cytotoxic activity has been further demonstrated by inducing a major loss of activity in an analogue after replacing highly hydrophobic residues with more hydrophilic ones.

Several myeloid antimicrobial peptide precursors have been shown to consist of a N‐terminal proregion similar to a protein named cathelin and a structurally varied C‐terminal antimicrobial domain. Proteins with these features have been named cathelicidins. In this paper we report the cDNA sequences of three ovine cathelicidins of 155, 160 and 190 residues, respectively, with cationic C‐terminal sequences corresponding to putative antimicrobial domains. These are structurally varied and include a Cys‐rich sequence of 12 residues, which is similar to the bovine antimicrobial cyclic dodecapeptide, a novel 29 residue sequence named SMAP‐29 with a possible α‐helical conformation, and a 60 residue sequence named Bac7.5, which appears to be a new member of the Pro‐ and Arg‐rich group of mammalian antimicrobial peptides.

The cdc2-family of serine/threonine kinases and their binding partners recently were implicated in developmental roles. We previously cloned a cdc2-related kinase, cdk9/PITALRE, that is able to phosphorylate the retinoblastoma protein in vitro. We describe here the cloning and the characterization of the mouse homolog of cdk9/PITALRE. The murine cDNA is 98% identical with humans and is expressed at high levels in brain and kidney tissues. The kinase activity and protein expression of cdk9/PITALRE were highest in terminally differentiated tissues such as the muscle and brain. In situ immunohistology and immunofluorescence detected cdk9/PITALRE protein not only within terminally differentiated cells such as muscle and neuronal cells, but also in proliferating cells. C2C12 and P19 cells induced to differentiate along muscle and neural lineages peaked in cdk9/PITALRE kinase activity at the end of differentiation. These results suggest that, among other roles, cdk9/PITALRE plays a role not unlike cdk5 in the differentiation of certain cell types. J. Cell. Physiol. 177:206–213, 1998. © 1998 Wiley-Liss, Inc.

Retinoblastoma (RB) family proteins pRb, p107 and pRb2/p130 are important cellular factors which play a well-recognized role as tumor and growth suppressors. These proteins are actively involved in the negative control of the cell cycle and their function is modulated via complex homeostatic processes, most of them involving post-translational regulation of their phosphorylation status. Interestingly, the family members p107 and pRb2/p130 share the ability to physically interact and inhibit the kinase activity of the Cdk2/Cyclin A and Cdk2/Cyclin E complexes. Regarding pRb2/p130, its inhibitory effect on the Cdk2/Cyclin A activity has been attributed to the “spacer” region. Recently, a 39 aa-long pRb2/p130 spacer-derived peptide (Spa310, aa 641-679) has been selected as the sequence responsible for Cdk2/Cyclin A inhibition. Following the identification of this active sequence, here we propose a computer-generated three-dimensional model of the interaction between the Cdk2/Cyclin A complex and the N-terminal 9-amino acid sequence of the Spa310 peptide. We believe this model as useful for the rational development of peptide or peptidomimetic kinase inhibitors to be used for the negative modulation of cell cycle in cancer cells.