Antitumor and antimetastatic activity of the immunoadjuvant peptidoglycan monomer PGM in mice bearing MCa mammary carcinoma

Summary The antitumor and antimetastatic activities of the water-soluble peptidoglycan monomer GlnNAc-Mur-NAc-L-Ala-D-iso-Gln-meso-diamminopimelic acid (-NH₂)-D-Ala-D-Ala (PGM), which has immunostimulant effects, have been evaluated in CBA mice bearing MCa mammary carcinoma. The antineoplastic effects of PGM depend strictly on the dosage and treatment schedule used. Though a significant inhibition of the primary tumor growth is observed over a wide range of dosage, only the IV administration of daily doses of 50 mg/kg/day on days 1, 5, 10, 15 inhibits spontaneous lung metastasis formation and in parallel prolongs the survival time of the treated mice. The magnitude of the antimetastatic effects of PGM depends on the degree of dissemination of the tumor, and is greater when the number of metastatic foci is low. Examination of the therapeutic potential of PGM in combination with surgery has further indicated that the timing of administration plays an important role in the overall effectiveness of this substance. A 5-day interval is necessary between two consecutive injections for the induction of significant increases of the survival times.     

NADPH oxidase 4 mediates upregulation of type 4 phosphodiesterases in human endothelial cells

The protective actions of prostacyclin (PGI(2) ) are mediated by cyclic AMP (cAMP) which is reduced by type 4 phosphodiesterases (PDE4) which hydrolyze cAMP. Superoxide (O2(-)) from NADPH oxidase (Nox) is associated with impaired PGI(2) bioactivity. The objective of this study, therefore, was to study the relationship between Nox and PDE4 expression in human umbilical vein endothelial cells (HUVECs). HUVECs were incubated with the thromboxane A(2) analog, U46619, 8-isoprostane F(2α) (8IP), or tumor necrosing factor alpha (TNFα) [±iloprost (a PGI(2) analog)] and the expression of PDE4A, B, C, and D and splice variants thereof assessed using Western blotting and qPCR and mRNA silencing of Nox4 and Nox5. Effects on cell replication and angiogenesis were also studied. U46619, 8IP, and TNFα increased the expression of Nox 4 and Nox 5 and all PDE4 isoforms as well as cell replication and tubule formation (index of angiogenesis), effects inhibited by mRNA silencing of Nox4 (but not Nox5) and iloprost and rolipram. These data demonstrate that upregulation of Nox4 leads to an upregulation of PDE4A, B, and D and increased hydrolysis of cAMP which in turn augments cell replication and angiogenesis. This mechanism may be central to vasculopathies associated with endothelial dysfunction since the PGI(2)-cAMP signaling axis plays a key role in mediating functions that include hemostasis and angiogenesis.     

Effect of different growth hormone dosages on the growth velocity in children born small for gestational age

To assess whether short-term growth hormone (GH) treatment can improve the linear growth in children who were born small for gestational age (SGA), we started a randomized multicenter trial in 26 age- and sex-matched prepubertal children born SGA. During the 1st year of GH therapy, all children received GH 0.23 mg/kg/week, then during the 2nd year, 13 children received the same dose (group A), and in the other 13 children, the dose of GH was doubled, i.e., 0.46 mg/kg/week (group B). During the 1st year of therapy, the growth velocity significantly (p<0.0001) increased in all patients. During the 2nd year, group A showed a significant decrease of the growth velocity (p<0.015), whereas group B maintained the growth rate. The height in group A children significantly increased during the 1st and the 2nd year of GH therapy (p<0.000002 and p<0.000001, respectively), reaching the normal range in 8 out of 13 children at the end of 2 years of GH therapy. The height in group B children significantly increased during the 1st and the 2nd year of GH therapy (p<0.000001 and p<0.000001, respectively), reaching the normal range in all 11 children who completed the GH therapy. The height gain was similar in groups A and B treated with the same GH dosage during the 1st year of therapy. A greater increase in height gain was found in children of group B treated with the higher GH dosage during the 2nd year of therapy as compared with group A (p<0.02). Significant increases in insulin-like growth factor I (p<0.0001), acid-labile subunit (p<0.0002), and bone/chronological age ratio (p<0.0001) were found after the 1st year of GH therapy, but no significant changes were observed during the 2nd year, independently of the GH dose. In conclusion, the height velocity of children born SGA significantly increases during the 1st year of GH therapy, diminishes, but can decrease during the 2nd year, if the GH dosage is not raised.     

Bacterial diversity and hydrography of Etoliko,an anoxic semi-enclosed coastal basin in Western Greece

Etoliko, an anoxic semi-enclosed basin, is part of a complex wetland in Western Greece extremely rich in biodiversity. It covers an area of 1,700 ha with an atypical orientation that has been formed tectonically. In order to identify the main factors influencing the bacterial profile at the Etoliko basin, 48 samples were collected, representing seasonal variation at four sampling stations. Physico-chemical analysis of the samples indicates the presence of three layers in the Etoliko basin: (1) low-density surface layer, (2) a layer with a steep density gradient, and (3) dense water below a depth of 20 m. A permanent halocline, whose thickness is varying seasonally, has been identified in the Etoliko basin water column, while the spatiotemporal salinity distribution was highly affected by the basin’s interaction with the nearby Messolonghi lagoon. The anoxic zone extends from 20 m below the surface to the bottom of the Etoliko basin in summer, while the bottom layer was hypoxic during winter. Bacterial populations were analyzed by Automated Ribosomal Intergenic Spacer Analysis (ARISA). Bacterial richness and diversity were calculated and compared across samples. Hierarchical analysis showed that ARISA clustered the surface water samples according to seasonal variation, while sediment and near-to-bottom water samples appear to be stable and to cluster together. Non-metric multidimensional scaling (MDS) indicates that bacterial composition depends on dissolved oxygen and salinity. Increase in salinity of the ecosystem leads to a significant reduction of the microbial diversity.     

Regulation of Smurf2 ubiquitin ligase activity by anchoring the E2 to the HECT domain

The conjugation of ubiquitin to proteins involves a cascade of activating (E1), conjugating (E2), and ubiquitin-ligating (E3) type enzymes that commonly signal protein destruction. In TGFbeta signaling the inhibitory protein Smad7 recruits Smurf2, an E3 of the C2-WW-HECT domain class, to the TGFbeta receptor complex to facilitate receptor degradation. Here, we demonstrate that the amino-terminal domain (NTD) of Smad7 stimulates Smurf activity by recruiting the E2, UbcH7, to the HECT domain. A 2.1 A resolution X-ray crystal structure of the Smurf2 HECT domain reveals that it has a suboptimal E2 binding pocket that could be optimized by mutagenesis to generate a HECT domain that functions independently of Smad7 and potently inhibits TGFbeta signaling. Thus, E2 enzyme recognition by an E3 HECT enzyme is not constitutively competent and provides a point of control for regulating the ubiquitin ligase activity through the action of auxiliary proteins.     

First draft genome sequencing of fennel (<Emphasis Type="Italic">Foeniculum vulgare</Emphasis> Mill.): identification of simple sequence repeats and their application in marker-assisted breeding

The development of F₁ hybrid varieties benefits from the synergistic effect of conventional and molecular marker-assisted breeding schemes. A sequencing run was carried out in Foeniculum vulgare (2n?=?2x?=?22) to develop the first genome draft and to identify microsatellites suitable for implementing multilocus SSR marker assays. A preliminary cytometric analysis allowed us to estimate the genome size (2C?=?2.64–2.86 pg), equal to about 1.34 Mbp for 1C genome, and to calculate the sequencing coverage (53×). The genome draft assembly into 300,408 scaffolds and its bioinformatic analysis enabled the annotation of coding and non-coding regions across the genome, including 103,306 SSR elements. A total of 100 microsatellites were randomly chosen among those with dinucleotide and trinucleotide repeat motifs and with a repeat motif length?≥?25 times and were preliminarily tested. Of these, 27 SSR markers, classified as suitable for genetic diversity analyses, were efficiently organized in five PCR multiplex assays and validated using a core collection of 100 fennel individuals potentially useful for the development of inbred lines and F₁ hybrids. All SSR loci were found to be polymorphic, scoring an observed number of marker alleles Na?=?207 and an average polymorphism information content PIC?=?0.69. The SSR data were used to calculate (i) the degree of homozygosity for the individual inbred lines (0.35?<?Ho?<?0.96), to eventually plan additional selfing or sibling cycles, and (ii) the degree of genetic similarity for all possible pair-wise comparisons between parental inbred lines (GS?=?0.55–0.77), to identify the most divergent combinations for the constitution of experimental F₁ hybrids. The integration of genotypic and phenotypic data was useful for implementing guidelines for precision hybrid breeding schemes in fennel.     

Sporophyte Ultrastructure inTortula muralis Hedw.

Summary The sporophyte ultrastructure ofTortula muralis Hedw. has been investigated by scanning and transmission electron microscopy. We have observed the twistings on the surface of the seta, the stomata in the lower part of the capsule and the disposition of cortical, conductive and parenchyma cells in the transversal and longitudinal sections of the seta. Moreover, in transection, the calyptra cells are thick-walled and present structures like tips in front of the epidermis cells of the capsule. Possible meanings of these morphological observations are discussed.     

Survival in different habitats: Extreme ultramafic and calcareous soils affect Stachys recta essential oils composition

Two populations of Stachys recta growing in Italy on ultramafic and calcareous soils have been studied for their essential oils. Although the yields were comparable, the composition of the essential oils differed significantly. Plants growing on ultramafic soil produced mainly non-terpene derivatives (55.7%), of which the most abundant ones were 1-octen-3-ol (38.2%) and (E)-3-hexen-1-ol (5.9%); the terpenes α-cadinol (6.1%) and δ-cadinene (5.6%) were also significantly represented. In contrast, the populations living on calcareous soil produced an essential oil dominated by terpenes (93.8%), with germacrene D (18.8%), β-caryophyllene (17.7%), 1,8-cineole (15.9%) and α-pinene (14.2%) among the main components.     

Weather-Related Flood and Landslide Damage: A Risk Index for Italian Regions

The frequency of natural hazards has been increasing in the last decades in Europe and specifically in Mediterranean regions due to climate change. For example heavy precipitation events can lead to disasters through the interaction with exposed and vulnerable people and natural systems. It is therefore necessary a prevention planning to preserve human health and to reduce economic losses. Prevention should mainly be carried out with more adequate land management, also supported by the development of an appropriate risk prediction tool based on weather forecasts. The main aim of this study is to investigate the relationship between weather types (WTs) and the frequency of floods and landslides that have caused damage to properties, personal injuries, or deaths in the Italian regions over recent decades. In particular, a specific risk index (WT-FLARI) for each WT was developed at national and regional scale. This study has identified a specific risk index associated with each weather type, calibrated for each Italian region and applicable to both annual and seasonal levels. The risk index represents the seasonal and annual vulnerability of each Italian region and indicates that additional preventive actions are necessary for some regions. The results of this study represent a good starting point towards the development of a tool to support policy-makers, local authorities and health agencies in planning actions, mainly in the medium to long term, aimed at the weather damage reduction that represents an important issue of the World Meteorological Organization mission.