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Objective To assess whether primary changes in endothelin-1 (ET-1) receptor responsiveness or secondary vessel functional modifications could characterize the effects evoked by ET-1 in the mesenteric vascular bed (MVB) of prehypertensive 5-week-old and 12-week-old spontaneously hypertensive rats (SHRs). Design and methods We used male 5-week-old and 12-week-old SHRs and sex- and age-matched Wistar-Kyoto (WKY) rats as controls. ET-1 receptor responsiveness was evaluated by ET-1 (0.04-2 μmol/l) concentration-response curves and repeated with indomethacin and BQ-123 (0.1-0.5 μmol/l), the latter a selective ET A receptor antagonist. ET B receptor responsiveness was tested by sarafotoxin S6c (1-100 nmol/l) and IRL-1620 (0.1-10 nmol/l) concentration-response curves, obtained in the noradrenaline-precontracted MVB. Results At 5 weeks of age, ET-1 induced a similar concentration-dependent contraction in SHRs and WKY rats, with an overlapping BQ-123 pA2 value (negative common logarithm of the antagonist that produces an agonist dose ratio of 2) in the two strains. Indomethacin was ineffective in both groups. Sarafotoxin S6c and IRL-1620 both evoked an ET B -mediated, significant relaxation, only in WKY rats. In 12-week-old SHRs, ET-1 evoked a markedly increased maximal effect compared with the response in WKY rats (P < 0.01); this was prevented by treatment with indomethacin. The BQ-123 pA2 value was higher in SHRs than in WKY rats (P < 0.01). Both sarafotoxin S6c and IRL-1620 evoked a significant concentration-dependent relaxation in WKY rats, which was not detected in SHR preparations. Conclusions Our results could suggest that the different responses evoked by ET-1 in the MVB of SHRs during the onset of hypertension may be related partially to primary alterations in the ET-1 receptorial pattern and partially to the onset of high blood pressure, leading to an impairment in the haemodynamic balance.

Abstract 4090 Transformation to acute myeloid leukemia (AML) is a known complication of MPN but the role of JAK2V617F mutation is still undefined. In 2006, Campbell described a possible model for the development of a JAK2WT-AML in a patient with JAK2V617F-MPN and more recently, Theocharides et al. reported that in up to 53% of the patients who developed secondary AML from a JAK2-mutated MPN the mutation was no longer detectable; however the results obtained with DNA extracted from cells scraped or laser-capture-microdissected from bone marrow (BM) or peripheral blood (PB) smears, were confirmed in fresh samples only in few cases. In this study, we collected, by cell sorting, blast cells and mature myeloid cells (granulocytes, GRA) from whole BM aspirates of 40 newly diagnosed patients with AML secondary to MPN (18 derived from PMF; 13 from PV and 9 from ET) and analyzed the JAK2 status before and after leukemic transformation in selected cell compartments. To evaluate the modification in the JAK2 status before and during leukemic transformation we performed ASO-PCR and (QRT)-PCR assay on total BM of the MPN phase and sorted cell populations from AML phase. At the time of MPN diagnosis, JAK2V617F was detectable in 28 of 40 patients (70%) (10 of 18 PMF; 12 of 13 PV and 6 of 9 ET). No cytogenetic abnormalities or MPL and JAK2-exone 12 mutations were detected at this stage. Median time to AML progression (TTP) was 5.09 years (yrs) (range 0.38 – 27.81). A significant difference (p= 0.02) in TTP was found grouping patients according to JAK2 status during the MPN phase [JAK2WT-MPN n=12, TTP median 15.10 yrs (0.38-16.32); JAK2 mutated-MPN n=28, TTP median 4.07 yrs (0.67-27.81)]. Eight patients showed additional abnormalities involving chromosomes 1, 5, 7, 8, 9, 12, 14, 17 and 20 while no other AML-associated mutations (FLT3, NPM, CEBPA, RUNX1) were detectable at this stage. In our cohort of patients we found that JAK2V617F mutation was still present at the blast transformation in both compartments: CD34+ cells (blasts) and CD15+ cells (GRA) in 24 of 28 JAK2 mutated MPN (86%). Four of 28 patients (14%) developed JAK2V617F negative AML starting from a mutated PV with a mean TTP of 5.14 yrs. Interestingly, the WT status was confirmed in blast cells but also in GRA. Surprisingly we found 2 cases of JAK2V617F mutated AML transforming from a WT-PMF. Also in this case the JAK2V617F positivity in the AML phase occurred in both GRA and blast compartments. No differences (p= 0.3) in the allele burden were found comparing MNCs from chronic phase with MNCs of leukemic transformations or comparing GRA with blasts in AML phase. In conclusion, these results contrast with the previous study in which the JAK2 mutation was lost in 53% of blasts during leukemia transformation. In our work, the loss of JAK2V617F mutation during AML progression is a rare event (14%). Additional studies in larger patient series and multivariate analysis are needed before a prognostic role of JAK2V617F mutation regarding time to leukemia transformation can be definitely assessed. Disclosures: Vannucchi: Novartis: Consultancy.

Objective The physiological and pathophysiological functions of endothelin-1 in modulating the regional blood flow of normal and spontaneously hypertensive rats (SHR) were studied in the perfused mesenteric vascular bed, a useful model for investigating resistance vessels. Design and methods We used 12-week-old SHR and Wistar-Kyoto (WKY) rats. Endothelin A (ET A ) receptor responsiveness was evaluated by endothelin-1 (0.2-2 μmol/l) concentration-response curves, and repeated in the presence of indomethacin and the ET A and endothelin B (ET B ) receptor antagonists BQ-485 and BQ-788, respectively. ET B receptor responsiveness was tested by sarafotoxin S6c concentration-response curves, obtained in the noradrenaline-precontracted mesenteric vascular bed, and repeated after treatment with BQ-788 and after endothelial denudation. Results In both groups, endothelin-1 induced concentration-dependent contraction; SHR exhibited a markedly increased maximal effect compared with WKY rats (P< 0.01). BQ-485 produced a shift to the right for endothelin-1 concentration-response curves in both groups, with a higher pA 2 (negative common logarithm of the antagonist that produces an agonist dose ratio of 2) value in SHR than in WKY rats (P< 0.01). The increase in the maximal effect produced by endothelin-1 in SHR was prevented by indomethacin, which also induced a significant increase in the endothelin-1 concentration producing the half-maximal response (EC 50 ) in SHR (P< 0.05). Sarafotoxin S6c produced an ET B -dependent endothelium-mediated relaxant effect in WKY rats, which was not observed in SHR. Conclusions The higher vasoconstriction induced by endothelin-1 in SHR may be related to a greater number of available ET A receptors, due to the presence of an ET A receptor subtype. This mechanism may be linked to the production of prostanoids that add to the direct endothelin-1-evoked vasoconstriction. These results, together with the lack of relaxation in response to sarafotoxin S6c in SHR, suggest that an imbalance in the endothelin-1 ability to induce both contraction and relaxation is present in SHR with sustained hypertension, manifesting as a greater contractile effect evoked in this strain.

The recombinant production of a novel chimeric polyprotein is described. The new protein contains either wild-type beta(2)-microglobulin (beta(2)m) or its truncated variant (DeltaN6 beta(2)m) (see picture). Structural characterization is achieved by means of single-molecule force spectroscopy studies of specific beta(2)m regions which could be involved in amyloidogenesis.

As modern electronics rapidly approach the ultimate level of integration (typically thought to be at the nanoscale level), the fascinating world of biomolecules provides new opportunities and directions for further miniaturization. In this work we review our results in the field of biomolecular electronics, starting from the fabrication of nanojunctions up to the implementation, of hybrid devices.

Background: Respiratory Syncytial Virus (RSV) is one of the main causes of respiratory infections during the first year of life. Very premature infants may contract more severe diseases and 'late preterm infants' may also be more susceptible to the infection. The aim of this study is to evaluate the risk factors for hospitalization during the first year of life in children born at different gestational ages in Italy. Methods: A cohort of 33-34 weeks gestational age (wGA) newborns matched by sex and age with two cohort of newborns born at 35-37 wGA and >37 wGA were enrolled in this study for a three-year period (2009-2012). Hospitalization for bronchiolitis (ICD-9 code 466.1) during the first year of life was assessed through phone interview at the end of the RSV season (November-March) and at the completion of the first year of life. Results: The study enrolled 2314 newborns, of which 2210 (95.5 %) had a one year follow-up and were included in the analysis; 120 (5.4 %) were hospitalized during the first year of life for bronchiolitis. Children born at 33-34 wGA had a higher hospitalization rate compared to the two other groups. The multivariate analysis carried out on the entire population associated the following factors with higher rates for bronchiolitis hospitalization: male gender; prenatal treatment with corticosteroids; prenatal exposure to maternal smoking; singleton delivery; respiratory diseases in neonatal period; surfactant therapy; lack of breastfeeding; siblings

Background Tobacco smoke exposure (TSE) is a worldwide health problem and it is considered a risk factor for pregnant women’s and children’s health, particularly for respiratory morbidity during the first year of life. Few significant birth cohort studies on the effect of prenatal TSE via passive and active maternal smoking on the development of severe bronchiolitis in early childhood have been carried out worldwide. Methods From November 2009 to December 2012, newborns born at ≥33 weeks of gestational age (wGA) were recruited in a longitudinal multi-center cohort study in Italy to investigate the effects of prenatal and postnatal TSE, among other risk factors, on bronchiolitis hospitalization and/or death during the first year of life. Results Two thousand two hundred ten newborns enrolled at birth were followed-up during their first year of life. Of these, 120 (5.4 %) were hospitalized for bronchiolitis. No enrolled infants died during the study period. Prenatal passive TSE and maternal active smoking of more than 15 cigarettes/daily are associated to a significant increase of the risk of offspring children hospitalization for bronchiolitis, with an adjHR of 3.5 (CI 1.5–8.1) and of 1.7 (CI 1.1–2.6) respectively. Conclusions These results confirm the detrimental effects of passive TSE and active heavy smoke during pregnancy for infants’ respiratory health, since the exposure significantly increases the risk of hospitalization for bronchiolitis in the first year of life. Electronic supplementary material The online version of this article (doi:10.1186/s12931-015-0312-5) contains supplementary material, which is available to authorized users.

Background Mutations of INF2 represent the major cause of familial autosomal dominant (AD) focal segmental glomerulosclerosis (FSGS). A few patients present neurological symptoms of Charcot-Marie-Tooth (CMT) disease but the prevalence of the association has not been assessed yet. Methods We screened 28 families with AD FSGS and identified 8 INF2 mutations in 9 families (32 patients overall), 3 of which were new. Mutations were in all cases localized in the diaphanous-inhibitory domain (DID) of the protein. Results Clinical features associated with INF2 mutations in our patient cohort included mild proteinuria (1.55 g/L; range 1-2.5) and haematuria as a unique symptom that was recognized at a median age of 21.75 years (range 8-30). Eighteen patients developed end-stage renal disease during their third decade of life; 12 patients presented a creatinine range between 1.2 and 1.5 mg/dL and 2 were healthy at 45 and 54 years of age. CMT was diagnosed in four cases (12.5%); one of these patients presented an already known mutation on exon 2 of INF2, whereas the other patients presented the same mutation on exon 4, a region that was not previously associated with CMT. Conclusions We confirmed the high incidence of INF2 mutations in families with AD FSGS. The clinical phenotype was mild at the onset of the disease, but evolution to ESRD was frequent. The incidence of CMT has, for the first time, been calculated here to be 12.5% of mutation carriers. Our findings support INF2 gene analysis in families in which renal failure and/or neuro-sensorial defects are inherited following an AD model.

Over the last millennia, the land between the Alps and the Mediterranean Sea, characterized by extraordinary habitat diversity, has seen an outstanding cross-cultural development. For the first time, this paper reports on the census of the Holocene archaeological sites that have been studied as part of archaeobotany in Italy (continental Italy, the Italian peninsula and islands) over the last quarter in a century. Pollen, non-pollen palynomorphs, seeds and fruits, woods/charcoals and other plant remains have all been analysed in multidisciplinary researches. A list of 630 sites has been provided by more than 15 archaeobotanical teams. The sites are located across the 20 regions of Italy, and in the Republic of San Marino (356 sites in northern Italy, 118 in central Italy, 156 in southern Italy and on the islands). They belong to several cultural phases: 321 sites are only pre-Roman, 264 are Roman/post-Roman, and 45 sites cover a broader range of time, present in both time spans. Site distribution is plotted in maps of site density according to geographical districts and the main chronological phases. The reference list helps to find analytical data referring to the descriptive papers that may be scattered throughout monographs and specific books on the matter.

Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is an autosomal recessive disorder characterized by ptosis and progressive external ophthalmoplegia, peripheral neuropathy, severe gastrointestinal dysmotility, cachexia, leukoencephalopathy and mitochondrial DNA depletion, multiple deletions, or both. This disorder is caused by loss-of-function mutations in the gene encoding thymidine phosphorylase (TP) a cytosolic enzyme that catalyzes phosphorolysis of thymidine to thymine and deoxyribose 1-phosphate. In MNGIE patients. TP activity is very low or absent resulting in dramatically elevated levels of plasma thymidine and deoxyuridine. TP is expressed in most human tissues but is not expressed in skeletal muscle usually affected in MNGIE suggesting that TP deficiency causes the disease through a toxic intermediate. In addition, TP is associated with angiogenesis and high concentrations of thymidine inhibit microvessels formation. In our preliminary study vessels number between two MINGIE patients and eleven controls was compared. Histologic slides were stained with Alkaline Phosphatase and ratio between blood vessels and fibres number was calculated for each sample. Even if cases and controls numbers are low and they have to be increased, a significative differerence between MINGIE and control patients was revealed suggesting that angiogenesis inhibition could be involved in MINGIE pathogenesis.

Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is an extremely rare autosomal recessive mitochondrial disease due to mutations to the nuclear TYMP gene. Thus, the mechanism which takes place in this condition leads to thymidine phosphorylase (TP) enzyme inactivity (either partial or complete - i.e., virtually lacking) resulting in toxic accumulation of thymidine and therefore altered mitochondrial DNA. MNGIE is characterized by severe gastrointestinal dysmotility, neurological impairment, poor quality of life and reduced life expectancy. Currently, therapeutic options for MNGIE are still limited. In order to restore TP activity, allogenic hematopoietic stem cell transplantation has been used as cellular source of TP although with modest results, the 5-year mortality rate being about 70%. This research tested the hypothesis that the liver can be an alternative source of TP. Eleven patients (7M; 35-55 years) who underwent hepatic resection for focal disorders were included. Margins of normal liver tissue were obtained during surgery and processed to identify, quantify and localize the TP protein by an array of techniques (Western Blot, ELISA, and immunohistochemistry) and to evaluate TYMP mRNA expression by qPCR. Western Blot identified TP in the liver with a TP/GAPDH ratio of 0.9±0.5. ELISA estimated TP content as 0.5±0.07 ng/mg of total protein. TP was identified in both nuclei and cytoplasm of hepatocytes and sinusoidal lining cells. TYMP mRNA was expressed in the liver. In this study, we provided evidence that the liver is an important source of TP. Our experiments represent a proof-of-concept that orthotopic liver transplantation (OLT) can be indicated as possible therapeutic alternative for MNGIE patients.

We report on the fabrication and single electron tunneling behaviour of large scale arrays of nanogap electrodes bridged by bisferrocene-gold nanoparticle hybrids (BFc-AuNP). Coulomb staircase was observed in the low temperature current-voltage curves measured on the junctions with asymmetric tunnel barriers. On the other hand, junctions with symmetric tunneling barrier exhibited mere nonlinear current voltage characteristics without discrete staircase. The experimental results agreed well with simulations based on the orthodox theory. The junction resistance showed thermally activated conduction behaviour at higher temperature. The overall voltage and temperature dependent results show that the transport behaviour of the large arrays of single particle devices obtained by a facile optical lithography and chemical etching process corresponds with the behaviour of single particle devices fabricated by other techniques like e-beam lithography and mechanical breaking methods.

Aims and background. Malignant mesothelioma is a fatal cancer of increasing incidence in north-eastern Italy. Together with asbestos, the polyomavirus SV40 was hypothesized to contribute to the onset of malignant mesothelioma. To investigate the putative role of SV40 in the individual susceptibility to asbestos-induced malignant mesothelioma, we conducted a molecular epidemiological study on a series of malignant mesothelioma patients from an area in north-eastern Italy hyperendemic for malignant pleural mesothelioma. Methods and study design. We collected 63 mesothelioma samples from incidence cases of patients diagnosed with malignant pleural mesothelioma in the period 2009-2010. DNA was extracted from patients’ tissue biopsies using the BioRobot EZ1 Qiagen workstation. SV40 sequence detection and quantification was performed by real time PCR females (61.5%), who showed significantly higher cancer co-morbidity (46.1% vs 12%, P = 0.005). SV40 was detected in 22% of malignant mesotheliomas, with a low viral load. In SV40-positive patients, a threefold increased risk of asbestos exposure was observed, more evident in females (OR 4.32) than in males (OR 1.20). Conclusions. Our findings indicate that a high prevalence of SV40 was present in malignant mesothelioma incident cases from an area hyperendemic for malignant mesothelioma in north-eastern Italy. Although asbestos is considered the main risk factor in malignant mesothelioma onset, a role for SV40 could be hypothesized.

We show that the combination of different electric fields in In0.5Ga0.5As/GaAs quantum-dot electroluminescent devices dramatically blueshifts the emission wavelength even though the photoluminescence occurs at the expected value of 1.3 μm at room temperature. Systematic photoluminescence (PL), electroluminescence (EL), and photocurrent measurements demonstrate that the electric field associated with the built-in dipole in the dots, directed from the base of the dots to their apex, and the device junction field lead to the depletion of the ground state. As a consequence, structures grown on n-type GaAs substrates exhibit electroluminescence only from the excited states (whereas the photoluminescence comes from the ground level). Instead, by growing the same device structure on p-type GaAs substrates, i.e., by reversing the direction of the built-in electric field of the device, the effect of the permanent dipole is strongly reduced, thus allowing us to obtain EL emission at the designed wavelength of 1.3 μ...

The crystal-structure, crystal-chemistry, and low-temperature behavior of a natural phillipsite-Na from the “Newer Volcanic Suite,” Richmond, Melbourne district, Victoria, Australia [K 0.75 (Na 0.88 Ca 0.57 ) ∑1.45 (Al 2.96 Ti 0.01 Si 5.07 ) ∑8.04 O 16 · 6.2H 2 O ( Z = 2), a = 9.9238(6), b = 14.3145(5), c = 8.7416(5) A, β = 124.920(9)°, and V = 1018.20(9) A 3 , space group P 2 1 / m ], have been investigated by means of in situ single-crystal X-ray diffraction, thermogravimetric analysis, and electron microprobe analysis in the wavelength dispersive mode. Two accurate structural refinements have been obtained on the basis of single-crystal X-ray diffraction data collected at 298 and 100 K, with: R 1 ( F ) 298K = 0.035, 3678 unique reflections with F o > 4σ( F o ) and 195 parameters, and R 1 ( F ) 100K = 0.035, 3855 unique reflections, F o > 4σ( F o ) and 195 parameters. In both refinements, the residuals in the final difference Fourier maps are e − /A 3 . A configuration of the extra-framework population different from that reported in previous studies is found at room temperature, with two possible sites for potassium (K1 and K2), one sodium/calcium site (Ca), and seven independent sites partially occupied by water molecules (W1, W2, W3, W4, W4′, W5, and W6). The low-temperature refinement shows that the framework component of the phillipsite structure is maintained within the T -range investigated. However, a change in the configuration of the extra-framework content occurs at low temperature: the occupancy of site K2 drastically decreases, while that of site K1 increases, the Ca site is split into two sub-sites (Ca1 and Ca2) and the number of water molecule sites decreases to six (W1, W2, W3, W4, W5, and W6). The rearrangement of the extra-framework population at low temperature is likely due to the change in shape (and size) of the micropores by tetrahedral tilting. The evolution of the “free diameters” with temperature shows that an “inversion” of the ellipticity of the eight-membered ring channel along [010] occurs. The evolution of the unit-cell parameters with T (measured at 298, 250, 200, 150, and 100 K) shows a continuous and linear trend, without evident thermo-elastic anomalies. The axial and volume thermal expansion coefficients (α j = l j −1 ·∂ l j /∂ T , α V = V −1 ·∂ V /∂ T ) between 100 and 298 K, calculated by weighted linear regression, yield the following values: α a = 1.8(1) × 10 −5 , α b = 1.2(1) × 10 −5 , α c = 1.1(1) × 10 −5 , and α V = 3.7(1) × 10 −5 K −1 . The thermal expansion of phillipsite is significantly anisotropic (α a :α b :α c = 1.64:1.09:1).

Two alternative pretreatment methods for depositing metal nanoparticles on mica for atomic force microscopy (AFM) imaging are presented. The treated substrates are flat and clean, thus they are amenable of use to characterize very small nanoparticles. The methods do not require any instrumentation or particular expertise. As they are also very quick, the need for storage of the prepared substrates is avoided altogether. These proposed methods, which are compared with the results of transmission electron microscopy analysis, allow the quick sizing and characterization of nanoparticles with the atomic force microscope and could thus help expanding the user community of nanoparticle researchers who could use the AFM for their characterization needs. Microsc. Res. Tech., 2008. © 2008 Wiley-Liss, Inc.

Il saggio ripercorre le trasformazioni dell'edilizia residenziale tra età tardoantica e altomedioevo, basandosi sulle testimonianze di fonti scritte ed archeologiche, con particolare riferimento alle tipologie e ai materiali costruttivi ed alle diverse esigenze della committenza. Individua, inoltre, i caratteri materiali delle residenze di IX-X secolo, parallelamente alle trasformazioni del tessuto sociale di un centro urbano di area romanica

Il saggio analizza le motivazioni e i criteri posti alla base della prima edizione a stampa degli Statuti cittadini modenesi del 1327 - unica redazione per il secolo XIV a noi pervenuta - realizzata nel 1864 per iniziativa della Deputazione di Storia Patria per le Provincie Modenesi (Aedes Muratoriana). L'obiettivo è quello di mettere a fuoco tale importante iniziativa editoriale nel contesto delle prospettive che alimentano il contemporaneo sviluppo della scienza storica in età postunitaria e degli specifici obiettivi perseguiti dalle neonate Deputazioni di Storia Patria e Società Storiche, alle quali è affidato il decisivo e delicato compito di recuperare le molteplici testimonianze su cui fondare l'identità e le radici della prima comunità nazionale italiana.