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•Trace-element analysis distinguishes between outcrops of Tufo del Palatino tuff.•Historical, archaeological, and geological evidence for ancient Capitoline quarry.•Stone used in Temple of Jupiter in Rome came from quarry at the Capitoline Hill.•Capitoline quarry potential source of stone for other Archaic monuments in Rome.

In this paper we present new geochemical and petrographic data aimed at identifying the volcanic materials used in Roman times to produce the fine aggregate of mortars employed in the construction of terrestrial (harenae fossiciae) and maritime (pulvis) masonry structures, described by Vitruvius in De Architectura. Using trace-element discrimination diagrams, this study establishes the lithological provenance of a selected dataset of mortar aggregates sampled from Roman edifices in the ancient Etruscan towns of Sutri, Vulci and Volsinii (Bolsena), from Rome (Temple of Castor and Pollux, Temple B of Largo Argentina, Forum of Trajan), and from the Roman harbors of Anxur(Terracina) and of Claudius(Fiumicino). We show that Vitruvius was referring to the loose pyroclastic products (pozzolan) erupted by the volcanoes of the Roman Magmatic Province, cropping out in Tuscany, Latium and Campania, that were selectively and systematically exploited to produce the aggregate in the different regions of central Italy. In particular, we identify the specific volcanic products occurring in northern Latium and in Rome from which the aggregates of the investigated mortars were exploited, and we correlate them to the four kinds of harenae fossiciaedefined by Vitruvius as carbunculus, rubra, nigraand cana. We also show that the material extracted from the mortars of the Roman ports sampled for this work, that Vitruvius calls pulvis, corresponds to the pumice-fall deposits erupted by the Phlegrean Fields volcanic district, and we discuss the archaeological implications and the historical background of the leadership held by the Roman town of Puteoli(Pozzuoli) in the construction technique and exportation of materials for maritime structures.

In this work, thermal degradation of commercial polymer mixtures was evaluated as a promising approach to the production of ceramic brake components. Currently, there is a wide interest for the production of cost effective ceramic pads, and carbon based materials are good candidates both in monolithic form and as porous networks to be reinfiltrated with a convenient polymer. The process parameters for obtaining porous glassy carbon, starting from mixtures of commercial polymers, have been evaluated in this work with the goal of producing samples with optimised mechanical properties. A preliminary study of the pyrolysis process has been carried out using thermogravimetric analysis coupled with mass spectrometry Raman spectroscopy and field emission scanning electron microscopy coupled with energy dispersive X-ray in order to define the feasibility of the method and to point out the critical issues. Mechanical tests have been performed in order to understand the relation between the carbonisation process and the mechanical properties.

Drug addiction results from the interplay between social and biological factors. Among these, genetic variables play a major role. The use of genetically related inbred rat strains that differ in their preference for drugs of abuse is one approach of great importance to explore genetic determinants. Lewis and Fischer 344 rats have been extensively studied and it has been shown that the Lewis strain is especially vulnerable to the addictive properties of several drugs when compared with the Fischer 344 strain. Here, we have used microarrays to analyze gene expression profiles in the frontal cortex and nucleus accumbens of Lewis and Fischer 344 rats. Our results show that only a very limited group of genes were differentially expressed in Lewis rats when compared with the Fischer 344 strain. The genes that were induced in the Lewis strain were related to oxygen transport, neurotransmitter processing and fatty acid metabolism. On the contrary genes that were repressed in Lewis rats were involved in physiological functions such as drug and proton transport, oligodendrocyte survival and lipid catabolism. These data might be useful for the identification of genes which could be potential markers of the vulnerability to the addictive properties of drugs of abuse.

Preliminary results on Pt‐based electrocatalysts prepared with CMK3 mesoporous carbons are presented. The electro‐catalytic performance towards the oxygen reduction reaction (ORR) was compared to that of commercial Pt/C‐Vulcan catalysts with the same Pt content. Polarisation tests were carried out with a single PEM fuel cell containing the Pt/OMC cathodes. The analysis of these tests, based on a semi‐empirical determination of the various polarisation terms, showed an interesting catalytic activity for ORR for the cathodes prepared with mesoporous carbon supports. The preliminary results indicate some critical aspects which need further investigation to improve the ohmic resistance limitation of these materials. Particularly, the variation of the grain size distribution with the Pt content and the effective distribution of Nafion® on the OMC surface should be considered.

We have tested a new kind of Fabry-Perot long-baseline optical resonator proposed to reduce the thermal noise sensitivity of gravitational wave interferometric detectors--the "mesa beam" cavity--whose flat top beam shape is achieved by means of an aspherical end mirror. We present the fundamental mode intensity pattern for this cavity and its distortion due to surface imperfections and tilt misalignments, and contrast the higher order mode patterns to the Gauss-Laguerre modes of a spherical mirror cavity. We discuss the effects of mirror tilts on cavity alignment and locking and present measurements of the mesa beam tilt sensitivity.

We report on a search for gravitational-wave bursts in data from the three LIGO interferometric detectors during their third science run. The search targets sub-second bursts in the frequency range 100-1100 Hz for which no waveform model is assumed and has a sensitivity in terms of the root-sum-square(rss) strain amplitude of hrss~ 10?20Hz?1/2. No gravitational-wave signals were detected in the eight days of analysed data.

The expression of many cellular genes is modulated by DNA methylation and histone acetylation. These processes can influence malignant cell transformation and are also responsible for the silencing of DNA constructs introduced into mammalian cells for therapeutic or research purposes. As a better understanding of these biological processes may contribute to the development of novel cancer treatments and to study the complex mechanisms regulating gene silencing, we established a cellular system suitable to dissect the mechanisms regulating DNA methylation and histone acetylation. For this purpose, we stably transfected the neuroblastoma cell line U87 with a cytomegalovirus promoter-driven reporter gene construct whose expression was analyzed following treatment with the DNA methylation inhibitor 5'-aza-2'-deoxycytidine or histone deacetylation inhibitor trichostatin A. Both substances reactivated the silenced cytomegalovirus promoter, but with different reaction kinetics. Furthermore, whereas the kinetics of reactivation by trichostatin A did not substantially change over the time range considered (5 days), reactivation induced by 5'-aza-2'-deoxycytidine showed profound differences between day 1 and longer time points. We showed that this effect is related to the down-regulation of DNA replication by 5'-aza-2'-deoxycytidine. Finally, we have shown that the simultaneous administration of trichostatin A and 5'-aza-2'-deoxycytidine results in reactivation of the CMV promoter according to a cooperative, not synergistic or additive, mechanism. In conclusion, our cellular system should represent a powerful tool to investigate the complex mechanisms regulating gene silencing and to identify new anticancer drugs.

The aim of the present work was to study the time course effects in levels of mRNA encoding N-methyl-d-aspartate receptor subunit 1 (NMDAR1) after long-term cocaine self-administration (1 mgkg injection) and its extinction using a yoked-box procedure. NMDAR1 content was measured by quantitative in situhybridization histochemistry in prefrontal cortex, caudate-putamen, nucleus accumbens, olfactory tubercle, and piriform cortex immediately after cessation of the last session of cocaine self-administration (Day 0) and 1, 5, and 10 days after the extinction period. The results show that long-term cocaine self-administration and its extinction alter NMDAR1 gene expression in these forebrain regions, and that the changes depend upon the brain region examined and the type of cocaine administration (contingent, noncontingent, and saline). Compared to saline and noncontingent cocaine administration, contingent cocaine produced an up-regulation in NMDAR1 gene expression on Day 0 in all the brain regions analyzed. NMDAR1 levels of contingent animals decreased progressively in the absence of cocaine, and the decrement persisted 10 days after the extinction of cocaine self-administration behavior in all the forebrain areas, with the exception of olfactory tubercle. In contrast, noncontingent cocaine administration did not produce any change in NMDAR1 gene expression on Day 0, and extinction resulted in an increase of NMDAR1 mRNA content on Days 1 and 5 and returned to control (saline) values on Day 10. These results suggest that an interaction between environmental stimuli and the pharmacological action of cocaine during drug self-administration and its extinction may represent an important factor in the regulation of cocaine effects on NMDAR1 gene expression.

The thermal behavior of a purified galena in a chlorine surrounding was determined. The kinetic of the chlorination of galena and the stoichiometry of this reaction between 723 to 973 K (450 to 700°C) were measured by thermogravimetry. The microstructural characterization of the solid residues were determined by SEM, EDXS and XRD. The ΔHvapof the produced lead chloride volatilization was obtained by the application of Clausius-Clapeyron equation considering the process in an equilibrium state.

A single radial haemolysis (SRH) for reovirus antibody determination was developed and compared to standard haemagglutination-inhibition (HI) and complement fixation (CF) techniques. SRH appeared more simple and sensitive than CF, but less sensitive and less specific than HI.

In order to assess the possible association of cytomegalovirus (CMV) with cerebral damage, 65 children aged between 2 and 10 years were tested for CMV isolation and antibody status. All of them had previously presented one or more episodes of febrile convulsions. At the time of our study, they were grouped according to their clinical features and electroencephalogram (EEG). 21 had typical and/or atypical absence attacks with 3 c/s spike-wave EEG (primary generalized epilepsy), 21 presented febrile seizures and spontaneous fits with focal EEG abnormalities, 23 had no seizures and a normal EEG. We performed the same investigation in 41 healthy children. The study showed a similar isolation rate of CMV in healthy subjects and in patients without neurological and EEG alterations, while a significantly higher isolation rate was observed in the groups with neurological and EEG abnormalities. As for serology, no significant difference was observed between the four groups, although the positivity rate of the healthy children was lower than in the other three groups. The study suggests a possible association of CMV with neurological and EEG abnormalities after febrile seizures.

The solution structure of the complex between 4', 6-diamidino-2-phenylindole (DAPI) and DNA oligomer (d(CGATCG))2 at a 2:1 drug/duplex ratio has been characterized by combined use of proton one- and two-dimensional NMR spectroscopy, molecular mechanics, and molecular dynamics computations. Intermolecular nuclear Overhauser effects (NOEs), DNA structure perturbations, and resonance shifts induced by binding provide evidence that DAPI interacts with DNA hexamer by two different binding mechanisms, in fast exchange on the NMR time scale, without any significant distortion of the B-type conformation of DNA hexamer. The results indicate that the ligand binds into the minor groove of the central 5'-ATC-3' region of the hexamer and on the outside of the oligomer by a pi,pi-stacking interaction with the terminal C1:G6 base pairs. A model for both binding mechanisms that accounts for all experimental data was generated by molecular mechanics and dynamics calculations based on experimental NOEs. In the minor groove binding, N2 amino group of G2 precludes a deep insertion of phenyl ring of DAPI into the groove. Position and orientation of the drug in the external stacking interaction resemble those suggested for intercalation of DAPI between C:G base pairs.

Somatic fusion of mesophyll protoplasts was used to produce hybrids between the frost-tolerant species Solanum commersonii (2n=2x=24) and dihaploid S. tuberosum (2n=2x=24). This is a sexually incompatible combination due to the difference in EBN (Endosperm Balance Number, Johnston et al. 1980). Species with different EBNs as a rule are sexually incompatible. Fifty-seven hybrids were analysed for variation in chromosome number, morphological traits, fertility and frost tolerance. About 70% of the hybrids were tetraploid, and 30% hexaploid. Chloroplast counts in stomatal guard cells revealed a low frequency of cytochimeras. The frequency of aneuploids was relatively higher at the hexaploid level (hypohexaploids) than at the tetraploid level (hypotetraploids). The somatic hybrids were much more vigorous than the parents, and showed an intermediate phenotype for several morphological traits and moderate to profuse flowering. Hexaploid hybrid clones were less vigorous and had a lower degree of flowering than the tetraploid hybrid clones. All of the hybrids were female fertile but male sterile except for one, which was fully fertile and self-compatible. Many seeds were produced on the latter clone by selfing and on the male-sterile clones by crossing. The somatic hybrid plants showed an introgression of genes for frost tolerance and an adaptability to cold from S. commersonii. Therefore, the use of these somatic hybrids in breeding for and in genetic esearch on frost tolerance and cold-hardening is suggested.

The Na+-independent binding of [3H]ß-alanine to rat brain stem plus spinal cord was reinvestigated, in order to study in more detail the characteristics of previously described ß-alanine binding processes. Binding was absent when amino acid-free postnuclear supernatants or crude synaptic membranes were used. Experiments performed with several other Na+-free preparations showed a sole binding component, irrespective of the preparation used. Biochemical characterization of this Na+-independent binding, using frozen/thawed/washed synaptosomal-mitochodrial fractions, showed that binding reached a plateau between 7 min and 13 min, increasing thereafter. Binding was linear with fraction protein over a range of 200–415 µg/ml incubation medium. Binding was completely inhibited by glycine, alanine, a-aminobutyric acid, ß-aminoisobutyric acid, hypotaurine and strychnine, and to a lesser extent by 2,2-dimethyl-ß-alanine, brucine and gelsemine. It was insensitive to taurine, ?-aminobutyric acid (GABA), 2-guanidinoethanesulfonic acid (GES), carnosine, and bicuculline methiodide. Binding was reversible, saturable (KD 20 µM), and heat sensitive.

We present the DNA sequence of a 6.7-kilobase member of the rat long interspersed repeated DNA family (LINE or L1Rn). This member (LINE 3) is flanked by a perfect 14-base-pair (bp) direct repeat and is a full-length, or close-to-full-length, member of this family. LINE 3 contains an approximately 100-bp A-rich right end, a number of long (greater than 400-bp) open reading frames, and a ca. 200-bp G + C-rich (ca. 60%) cluster near each terminus. Comparison of the LINE 3 sequence with the sequence of about one-half of another member, which we also present, as well as restriction enzyme analysis of the genomic copies of this family, indicates that in length and overall structure LINE 3 is quite typical of the 40,000 or so other genomic members of this family which would account for as much as 10% of the rat genome. Therefore, the rat LINE family is relatively homogeneous, which contrasts with the heterogeneous LINE families in primates and mice. Transcripts corresponding to the entire LINE sequence are abundant in the nuclear RNA of rat liver. The characteristics of the rat LINE family are discussed with respect to the possible function and evolution of this family of DNA sequences.

The insulin 1, but not the insulin 2, locus is polymorphic (i.e., exhibits allelic variation) in rats. Restriction enzyme analysis and hybridization studies showed that the polymorphic region is 2.2 kilobases upstream of the insulin 1 coding region and is due to the presence or absence of an approximately 2.7-kilobase repeated DNA element. DNA sequence determination showed that this DNA element is a member of a long interspersed repeated DNA family (LINE) that is highly repeated (greater than 50,000 copies) and highly transcribed in the rat. Although the presence or absence of LINE sequences at the insulin 1 locus occurs in both the homozygous and heterozygous states, LINE-containing insulin 1 alleles are more prevalent in the rat population than are alleles without LINEs. Restriction enzyme analysis of the LINE-containing alleles indicated that at least two versions of the LINE sequence may be present at the insulin 1 locus in different rats. Either repeated transposition of LINE sequences or gene conversion between the resident insulin 1 LINE and other sequences in the genome are possible explanations for this.

AimTo evaluate the diagnostic value of telomerase expression in intracranial tumours.Methods98 surgical specimens from different neoplasms were analysed by the telomeric repeat amplification protocol (TRAP) and the presence of telomerase compared with the histological diagnosis and the proliferation index.ResultsA high degree of positivity for telomerase activity was found in glioblastomas and atypical/anaplastic meningiomas. Telomerase activity was poorly detected in anaplastic astrocytomas.ConclusionsThe TRAP assay seems to be a valuable index for identifying meningeal tumours with aggressive behaviour.

AimTo evaluate the diagnostic value of telomerase expression in intracranial tumours.Methods98 surgical specimens from different neoplasms were analysed by the telomeric repeat amplification protocol (TRAP) and the presence of telomerase compared with the histological diagnosis and the proliferation index.ResultsA high degree of positivity for telomerase activity was found in glioblastomas and atypical/anaplastic meningiomas. Telomerase activity was poorly detected in anaplastic astrocytomas.ConclusionsThe TRAP assay seems to be a valuable index for identifying meningeal tumours with aggressive behaviour.

Abstract: We describe a novel double nucleotide substitution in the SRY gene of a 46,XY female with gonadal dysgenesis or Swyer syndrome. The SRY sequence was analysed by both the single-strand conformational polymorphism assay and direct DNA sequencing of products from the polymerase chain reaction. A double nucleotide substitution was identified at codon 18 of the conserved HMG box motif, causing an arginine to asparagine amino-acid substitution. The altered residue is situated in the high mobility group (HMG)-related box of the SRY protein, a potential DNA-binding domain. Since the mutation abolishes one HhaI recognition site, the results were confirmed by HhaI restriction mapping. No other mutations were found in the remaining regions of the gene. The corresponding DNA region from the patient’s brother was analysed and found to be normal. We conclude that the SRY mutation in the reported XY female occurred de novo and is associated with sex reversal.

Telomeric Repeat Amplification Protocol (TRAP) is a sensitive procedure to measure telomerase activity in small samples of cell or tissue extracts. Due to the strict correlation between high levels of telomerase activity and neoplastic transformation, TRAP assay could provide an important diagnostic marker of malignancy. Although the original TRAP assay is very sensitive and some improvements have been described, occasional artifacts still persist in the modified procedures. Here we describe how changes in the sequence of the primer used for the amplification step enhance the reproducibility and sensitivity in the TRAP assay.

A typical G-rich telomeric DNA strand, which runs 5'-->3' toward the chromosome ends, protrudes by several nucleotides in lower eukaryotes. In human chromosomes long G-rich 3'-overhangs have been found. Apart from the standard G-rich tail, several non-canonical terminal structures have been proposed. However, the mechanism of long-tail formation, the presence and the role of these structures in telomere maintenance or shortening are not completely understood. In a search for a simple method to accurately measure the 3'-overhang we have established a protocol based on the ligation of telomeric oligonucleotide hybridized to non-denatured DNA under stringent conditions (oligonucleotide ligation assay with telomeric repeat oligonucleotide). This method enabled us to detect a large proportion of G-rich single-stranded telomeric DNA that was as short as 24 nt. Nevertheless, we showed G-tails longer than 400 nt. In all tested cells the lengths ranging from 108 to 270 nt represented only 37% of the whole molecule population, while 56-62% were <90 nt. Our protocol provides a simple and sensitive method for measuring the length of naturally occurring unpaired repeated DNA.

A typical G-rich telomeric DNA strand, which runs 5′→3′ toward the chromosome ends, protrudes by several nucleotides in lower eukaryotes. In human chromosomes long G-rich 3′-overhangs have been found. Apart from the standard G-rich tail, several non-canonical terminal structures have been proposed. However, the mechanism of long-tail formation, the presence and the role of these structures in telomere maintenance or shortening are not completely understood. In a search for a simple method to accurately measure the 3′-overhang we have established a protocol based on the ligation of telomeric oligonucleotide hybridized to non-denatured DNA under stringent conditions (oligonucleotide ligation assay with telomeric repeat oligonucleotide). This method enabled us to detect a large proportion of G-rich single-stranded telomeric DNA that was as short as 24 nt. Nevertheless, we showed G-tails longer than 400 nt. In all tested cells the lengths ranging from 108 to 270 nt represented only 37% of the whole molecule population, while 56-62% were &lt;90 nt. Our protocol provides a simple and sensitive method for measuring the length of naturally occurring unpaired repeated DNA.