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Sample records for gli iperstrati sferici

  1. Radiation-induced chemical evolution of glycine to (Gly)2, (Gly)3, and (Gly)4

    Matsui, T.; Izumi, Y.; Kamohara, M.; Nakagawa, K.; Yokoya, A.

    2006-01-01

    Recently amino acids were detected from some meteorites. Since these amino acids were found after hydrolysis, some oligopeptides were possibly formed in space. A simulation experiment of chemical evolution from Glycine (Gly) to Glycylglycine ((Gly)2) was reported by Kaneko et al. In this work, we irradiated (Gly)2 with 8 eV vacuum ultraviolet photons or with 530 eV soft X-ray photons and examined absolute values of quantum yield of radiation-induced chemical evolution from Gly2 to Glycylglycylglycine ((Gly)3) and Glycylglycylglycylglycine ((Gly)4). Thin films of (Gly)2 were prepared on quartz plate or CuBe plate with a vacuum evaporation technique. These samples were irradiated by 8 eV photons from a Xe 2 * excimer lamp or by 530 eV soft X-ray photons at SPring-8 Synchrotron Radiation Facility. Irradiated samples were analyzed with a high performance liquid chromatography HPLC. Decomposition of (Gly)2 and production of Gly, (Gly)3 and (Gly)4 were observed. Quantum yield Y was defined to be N = Y N 0 , where N is the number of produced or decomposed molecule, and N 0 is the number of (Gly)2 molecules excited by photons. Obtained results by 8 eV irradiation were summarized in Table 1. The similar magnitude of decomposition of (Gly)2 may show that yield of the primary breaking reaction upon photo-excitation is of similar magnitude. It should be noted that (Gly)3 and (Gly)4 was produced by irradiation with the yield of 10 -4 without any catalysis. For soft X-ray irradiation, yield of Gly was tentatively determined to be about 40. This largervalue than that for 8 eV irradiation may originate from large energy of incident soft X-ray photons just like a result reported by Simakov et al. We will discuss in detail at the conference. (authors)

  2. Gli atomi di Boltzmann

    Lindley, David

    2002-01-01

    Ludwig Boltzmann (1844-1906) è il fisico e matematico austriaco che negli ultimi decenni dell'Ottocento e ancora ai primi del Novecento lottò contro l'opinione dominante tra gli scienziati dell'epoca per affermare la teoria atomica della materia. È noto come con Albert Einstein e fino a oggi la fisica si sia sviluppata e abbia celebrato i propri trionfi lungo le linee anticipate da Boltzmann. La controversia con Mach non riguardava soltanto l'esistenza degli atomi, ma l'intero modo di fare fisica che Boltzmann non riteneva di dover limitare allo studio di quantità misurabili, introducendo invece spiegazioni più elaborate basate su ipotesi più ampie.

  3. Surface-enhanced Raman spectrum of Gly-Gly adsorbed on the silver colloidal surface

    Xiaojuan, Yuan; Huaimin, Gu; Jiwei, Wu

    2010-08-01

    Raman and SERS spectra of homodipeptide Gly-Gly and Gly were recorded and compared in this paper, and band assignment for the functional groups contained in these molecules was analyzed in detail. Time-dependent and pH-dependent SERS spectra of Gly-Gly molecule adsorbed on nano-colloidal silver surface were also studied. The time-dependent SERS spectra of Gly-Gly are characterized by the increase in intensity of bands primarily representing the vibrational signatures emanating from the amino and amide moiety of Gly-Gly molecule. It is found that the adsorption style of Gly-Gly on the silver colloid changes as time goes on; at 5 min after adding the sample to the silver colloid, Gly-Gly adsorbs on silver surface firstly through the carboxylate, amino and amide groups, and then the carboxylate group is far away from the silver surface at 10 min to 3 days. The SERS variation of Gly-Gly with the change of pH suggests that the adsorption style is pH-dependent, the different adsorption behavior of the Gly-Gly occurs on silver surface at different pH values.

  4. Domestic allergens; Gli allergeni domestici

    Bressa, G. [Padua Univ., Padua (Italy). DIpt. di Tossicologia Ambientale

    2000-12-01

    Closed rooms can be the ideal habitat for the growth of many micro-organisms, some of which pathogenic for man. Indoor biologic contamination sources can vary: 1) external air can, through dust particles or water droplets, transfer a large number of germs inside the house; 2) air conditioning plants can become, if maintenance isn't carried out at regular intervals, a cultural medium for microbes which later spread to the air of house or office; 3) the presence of sick people or healthy carriers of pathogenic germs in closed rooms can represent a significant source of biologic contamination; 4) lastly, pets too can be considered as disease carriers. Particularly interesting is the so-called biologic dust - where mites proliferate - which, being trapped in curtains, carpeting, tapestry, fabrics and carpets, often causes respiratory system diseases. [Italian] Gli ambienti chiusi possono costituire l'habita ideale per la crescita di molti microorganismi, alcuni dei quali patogeni per l'uomo. Le fonti di contaminaizone biologica indoor possono essere diverse: 1) l'aria esterna puo' trasferire all'interno, attraverso particelle di polvere o goccioline d'acqua, un gran numero di germi; 2) gli impianti di condizionamento possono divenire, quando non si effettua una corretta manutenzione periodica, terreno di coltura di microbi che successivamente si diffondono nell'aria dell'abitazione o dell'ufficio; 3) la presenza in locali chiusi di persone malate o portatrici sane di germi patogeni puo' costituire un'importante sorgente di contaminazione biologica; 4) infine anche gli animali domestici possono essere considerati vettori di malattie. Di particolare interesse sono le polveri cosidette biologiche, in cui proliferano gli acari, che, essendo trattenute da tendaggi, parati, moquette, tessuti e tappeti, provocano frequentemente malattie dell'apparato respiratorio.

  5. Differential requirements for Gli2 and Gli3 in the regional specification of the mouse hypothalamus

    Roberta eHaddad-Tóvolli

    2015-03-01

    Full Text Available Secreted protein Sonic hedgehog (Shh ventralizes the neural tube by modulating the crucial balance between activating and repressing functions (GliA, GliR of transcription factors Gli2 and Gli3. This balance—the Shh-Gli code—is species- and context-dependent and has been elucidated for the mouse spinal cord. The hypothalamus, a forebrain region regulating vital functions like homeostasis and hormone secretion, shows dynamic and intricate Shh expression as well as complex regional differentiation. Here we asked if particular combinations of Gli2 and Gli3 and of GliA and GliR functions contribute to the variety of hypothalamic regions, i.e. we wanted to clarify the hypothalamic version of the Shh-Gli code. Based on mouse mutant analysis, we show that: 1 hypothalamic regional heterogeneity is based in part on differentially stringent requirements for Gli2 or Gli3; 2 another source of diversity are differential requirements for Shh of neural vs non-neural origin; 3 Gli2 is indispensable for the specification of a medial progenitor domain generating several essential hypothalamic nuclei plus the pituitary and median eminence; 4 the suppression of Gli3R by neural and non-neural Shh is essential for hypothalamic specification. Finally, we have mapped our results on a recent model which considers the hypothalamus as a transverse region with alar and basal portions. Our data confirm the model and are explained by it.

  6. Secondary Structure Adopted by the Gly-Gly-X Repetitive Regions of Dragline Spider Silk

    Geoffrey M. Gray

    2016-12-01

    Full Text Available Solid-state NMR and molecular dynamics (MD simulations are presented to help elucidate the molecular secondary structure of poly(Gly-Gly-X, which is one of the most common structural repetitive motifs found in orb-weaving dragline spider silk proteins. The combination of NMR and computational experiments provides insight into the molecular secondary structure of poly(Gly-Gly-X segments and provides further support that these regions are disordered and primarily non-β-sheet. Furthermore, the combination of NMR and MD simulations illustrate the possibility for several secondary structural elements in the poly(Gly-Gly-X regions of dragline silks, including β-turns, 310-helicies, and coil structures with a negligible population of α-helix observed.

  7. Role of GLI2 in hypopituitarism phenotype.

    Arnhold, Ivo J P; França, Marcela M; Carvalho, Luciani R; Mendonca, Berenice B; Jorge, Alexander A L

    2015-06-01

    GLI2 is a zinc-finger transcription factor involved in the Sonic Hedgehog pathway. Gli2 mutant mice have hypoplastic anterior and absent posterior pituitary glands. We reviewed the literature for patients with hypopituitarism and alterations in GLI2. Twenty-five patients (16 families) had heterozygous truncating mutations, and the phenotype frequently included GH deficiency, a small anterior pituitary lobe and an ectopic/undescended posterior pituitary lobe on magnetic resonance imaging and postaxial polydactyly. The inheritance pattern was autosomal dominant with incomplete penetrance and variable expressivity. The mutation was frequently inherited from an asymptomatic parent. Eleven patients had heterozygous non-synonymous GLI2 variants that were classified as variants of unknown significance, because they were either absent from or had a frequency lower than 0.001 in the databases. In these patients, the posterior pituitary was also ectopic, but none had polydactyly. A third group of variants found in patients with hypopituitarism were considered benign because their frequency was ≥ 0.001 in the databases. GLI2 is a large and polymorphic gene, and sequencing may identify variants whose interpretation may be difficult. Incomplete penetrance implies in the participation of other genetic and/or environmental factors. An interaction between Gli2 mutations and prenatal ethanol exposure has been demonstrated in mice dysmorphology. In conclusion, a relatively high frequency of GLI2 mutations and variants were identified in patients with congenital GH deficiency without other brain defects, and most of these patients presented with combined pituitary hormone deficiency and an ectopic posterior pituitary lobe. Future studies may clarify the relative role and frequency of GLI2 alterations in the aetiology of hypopituitarism. © 2015 Society for Endocrinology.

  8. Radiation Chemical Studies of Gly-Met-Gly in Aqueous Solution

    Barata-Vallejo, Sebastian; Ferreri, Carla; Zhang, Tao; Permentier, Hjalmar; Bischoff, Rainer; Bobrowski, Krzysztof; Chatgilialoglu, Chryssostomos

    2016-01-01

    Important biological consequences are related to the reaction of HO(•) radicals with methionine (Met). Several fundamental aspects remain to be defined when Met is an amino acid residue incorporated in the interior of peptides and proteins. The present study focuses on Gly-Met-Gly, the simplest

  9. Gli as a novel therapeutic target in malignant pleural mesothelioma.

    Hui Li

    Full Text Available Malignant pleural mesothelioma (MPM is a highly aggressive tumor with poor prognosis. Current treatment is rarely curative, thus novel meaningful therapies are urgently needed. Inhibition of Hedgehog (Hh signaling at the cell membrane level in several cancers has shown anti-cancer activity in recent clinical studies. Evidence of Hh-independent Gli activation suggests Gli as a more potent therapeutic target. The current study is aimed to evaluate the potential of Gli as a therapeutic target to treat MPM. The expression profiles of Gli factors and other Hh signaling components were characterized in 46 MPM patient tissue samples by RT-PCR and immunohistochemistry. Cultured cell lines were employed to investigate the requirement of Gli activation in tumor cell growth by inhibiting Gli through siRNA or a novel small molecule Gli inhibitor (Gli-I. A xenograft model was used to evaluate Gli-I in vivo. In addition, a side by side comparison between Gli and Smoothened (Smo inhibition was conducted in vitro using siRNA and small molecule inhibitors. Our study reported aberrant Gli1 and Gli2 activation in a large majority of tissues. Inhibition of Gli by siRNAs or Gli-I suppressed cell growth dramatically both in vitro and in vivo. Inhibition of Gli exhibited better cytotoxicity than that of Smo by siRNA and small molecule inhibitors vismodegib and cyclopamine. Combination of Gli-I and pemetrexed, as well as Gli-I and vismodegib demonstrated synergistic effects in suppression of MPM proliferation in vitro. In summary, Gli activation plays a critical role in MPM. Inhibition of Gli function holds strong potential to become a novel, clinically effective approach to treat MPM.

  10. The first N-terminal unprotected (Gly-Aib)n peptide: H-Gly-Aib-Gly-Aib-OtBu.

    Gessmann, Renate; Brückner, Hans; Petratos, Kyriacos

    2015-12-01

    Glycine (Gly) is incorporated in roughly half of all known peptaibiotic (nonribosomally biosynthesized antibiotic peptides of fungal origin) sequences and is the residue with the greatest conformational flexibility. The conformational space of Aib (α-aminoisobutyric acid) is severely restricted by the second methyl group attached to the Cα atom. Most of the crystal structures containing Aib are N-terminal protected. Deprotection of the N- or C-terminus of peptides may alter the hydrogen-bonding scheme and/or the structure and may facilitate crystallization. The structure reported here for glycyl-α-aminoisobutyrylglycyl-α-aminoisobutyric acid tert-butyl ester, C16H30N4O5, describes the first N-terminal-unprotected (Gly-Aib)n peptide. The achiral peptide could form an intramolecular hydrogen bond between the C=O group of Gly1 and the N-H group of Aib4. This hydrogen bond is found in all tetrapeptides and N-terminal-protected tripeptides containing Aib, apart from one exception. In the present work, this hydrogen bond is not observed (N...O = 5.88 Å). Instead, every molecule is hydrogen bonded to six other symmetry-related molecules with a total of eight hydrogen bonds per molecule. The backbone conformation starts in the right-handed helical region (and the left-handed helical region for the inverted molecule) and reverses the screw sense in the last two residues.

  11. Glycine transporter GlyT1, but not GlyT2, is expressed in rat dorsal root ganglion--Possible implications for neuropathic pain

    Schlösser, Lukas; Barthel, Franziska; Brandenburger, Timo; Neumann, Elena; Bauer, Inge; Eulenburg, Volker; Werdehausen, Robert; Hermanns, Henning

    2015-01-01

    Glycinergic inhibitory neurotransmission plays a pivotal role in the development of neuropathic pain. The glycine concentration in the synaptic cleft is controlled by the glycine transporters GlyT1 and GlyT2. GlyT1 is expressed throughout the central nervous system, while GlyT2 is exclusively

  12. Gli studi di Enrico Cerulli su Dante

    Andrea Celli

    2013-12-01

    Full Text Available Nel 1949 lo studioso di lingue semitiche Enrico Cerulli pubblicò un volume, intitolato Il “Libro della scala” e la questione delle fonti arabo-spagnole della Divina Commedia, che segnò gli studi danteschi, per lo meno per quanto riguarda il dibattito sulle fonti arabo-islamiche del medioevo europeo. Le ricerche di Cerulli su Dante si presentavano come una analisi di fatti di trasmissione culturale. Ma l’autore si era già affermato, nella prima parte della sua carriera, in un più controverso ambito di competenze: alto funzionario dell’amministrazione coloniale italiana e vice-governatore dell’Africa Orientale Italiana, è stato rilevante studioso di lingue del Corno d’Africa, cioè le lingue delle colonie italiane. Questo saggio vuole connettere gli studi danteschi di Cerulli alle responsabilità coloniali e post-coloniali dell’autore, attraverso la ricostruzione della sua biografia intellettuale. Difatti, da queste ricerche Cerulli ricavò il concetto di unità della civiltà mediterranea, formula di evidente valore politico in rapporto all’Italia del secondo dopoguerra. Il saggio prova a offrire anche uno stato dell’arte del dibattito attuale sulle fonti arabe di Dante.  In 1949, Enrico Cerulli, an Italian scholar of Ethiopic-Semitic languages, published a volume entitled The “Book of the Ladder” and the Problem of Divine Comedy’s Arabic-Spanish Sources, which was to become a landmark in the discussion on the knowledge held by medieval Europe of Arabic-Islamic culture. His investigation was an analysis of cases of cultural transmission. However, Cerulli had established himself during the first half of his career in a more controversial domain: he was a senior official of Italian colonial administration and he was already well known as an expert of Italian colonies’ languages and traditions. This essay, through a survey of Cerulli’s intellectual biography, aims to bridge his research on Dante and his involvement

  13. Gli2 activator function in preosteoblasts is sufficient to mediate Ihh-dependent osteoblast differentiation, whereas the repressor function of Gli2 is dispensable for endochondral ossification.

    Kesper, Dörthe Andrea; Didt-Koziel, Lydia; Vortkamp, Andrea

    2010-06-01

    Signaling of Indian hedgehog (Ihh), one of the key regulators of endochondral ossification is mediated by transcription factors of the Gli family, Gli1, Gli2, and Gli3. Gli3 and to a lesser extent Gli2 can be proteolytically processed into short repressor proteins. Upon Ihh signaling, processing is inhibited and the full-length proteins function as activators of transcription. Gli3 has been shown to mainly act as a repressor of Ihh target genes in chondrocytes, but the role of other Gli isoforms is less clear. Analyzing mouse mutants deficient for Ihh;Gli2 or Gli3;Gli2, we show here that the Gli2 repressor has no detectable function in chondrocyte or osteoblast differentiation. Instead, Gli2 seems to act as an activator to fully induce the expression of Ihh target genes in skeletal tissues. Furthermore, we show that, in the absence of Gli3, the activator function of Gli2 is sufficient to induce Ihh-dependent osteoblast differentiation.

  14. The position of the Gly-xxx-Gly motif in transmembrane segments modulates dimer affinity.

    Johnson, Rachel M; Rath, Arianna; Deber, Charles M

    2006-12-01

    Although the intrinsic low solubility of membrane proteins presents challenges to their high-resolution structure determination, insight into the amino acid sequence features and forces that stabilize their folds has been provided through study of sequence-dependent helix-helix interactions between single transmembrane (TM) helices. While the stability of helix-helix partnerships mediated by the Gly-xxx-Gly (GG4) motif is known to be generally modulated by distal interfacial residues, it has not been established whether the position of this motif, with respect to the ends of a given TM segment, affects dimer affinity. Here we examine the relationship between motif position and affinity in the homodimers of 2 single-spanning membrane protein TM sequences: glycophorin A (GpA) and bacteriophage M13 coat protein (MCP). Using the TOXCAT assay for dimer affinity on a series of GpA and MCP TM segments that have been modified with either 4 Leu residues at each end or with 8 Leu residues at the N-terminal end, we show that in each protein, centrally located GG4 motifs are capable of stronger helix-helix interactions than those proximal to TM helix ends, even when surrounding interfacial residues are maintained. The relative importance of GG4 motifs in stabilizing helix-helix interactions therefore must be considered not only in its specific residue context but also in terms of the location of the interactive surface relative to the N and C termini of alpha-helical TM segments.

  15. GLI3 Links Environmental Arsenic Exposure and Human Fetal Growth

    Emily F. Winterbottom

    2015-06-01

    Full Text Available Although considerable evidence suggests that in utero arsenic exposure affects children's health, these data are mainly from areas of the world where groundwater arsenic levels far exceed the World Health Organization limit of 10 μg/L. We, and others, have found that more common levels of in utero arsenic exposure may also impact children's health. However, the underlying molecular mechanisms are poorly understood. To address this issue, we analyzed the expression of key developmental genes in fetal placenta in a birth cohort of women using unregulated water supplies in a US region with elevated groundwater arsenic. We identified several genes whose expression associated with maternal arsenic exposure in a fetal sex-specific manner. In particular, expression of the HEDGEHOG pathway component, GLI3, in female placentae was both negatively associated with arsenic exposure and positively associated with infant birth weight. This suggests that modulation of GLI3 in the fetal placenta, and perhaps in other fetal tissues, contributes to arsenic's detrimental effects on fetal growth. We showed previously that arsenic-exposed NIH3T3 cells have reduced GLI3 repressor protein. Together, these studies identify GLI3 as a key signaling node that is affected by arsenic, mediating a subset of its effects on developmental signaling and fetal health.

  16. Complementary Gli activity mediates early patterning of the mouse visual system.

    Furimsky, Marosh; Wallace, Valerie A

    2006-03-01

    The Sonic hedgehog (Shh) signaling pathway plays a key role in the development of the vertebrate central nervous system, including the eye. This pathway is mediated by the Gli transcription factors (Gli1, Gli2, and Gli3) that differentially activate and repress the expression of specific downstream target genes. In this study, we investigated the roles of the three vertebrate Glis in mediating midline Shh signaling in early ocular development. We examined the ocular phenotypes of Shh and Gli combination mutant mouse embryos and monitored proximodistal and dorsoventral patterning by the expression of specific eye development regulatory genes using in situ hybridization. We show that midline Shh signaling relieves the repressor activity of Gli3 adjacent to the midline and then promotes eye pattern formation through the nonredundant activities of all three Gli proteins. Gli3, in particular, is required to specify the dorsal optic stalk and to define the boundary between the optic stalk and the optic cup.

  17. A Novel Gli3 Enhancer Controls the Gli3 Spatiotemporal Expression Pattern through a TALE Homeodomain Protein Binding Site ▿‡

    Coy, Sarah; Caamaño, Jorge H.; Carvajal, Jaime; Cleary, Michael L.; Borycki, Anne-Gaëlle

    2011-01-01

    The zinc finger transcription factor Gli3 is an essential mediator of hedgehog signaling. Gli3 has a dynamic expression pattern during embryonic development. In the neural tube, Gli3 transcripts are patterned along the anteroposterior and dorsoventral axes such that the initial broad expression in the posterior neural tube becomes dorsally restricted as neurogenesis takes place. Little is known about the molecular mechanisms that regulate this dynamic expression. Here, we report on a phylogen...

  18. Evolution and Functional Diversification of the GLI Family of Transcription Factors in Vertebrates

    Amir Ali Abbasi

    2009-05-01

    Full Text Available Background: In vertebrates the “SONIC HEDGEHOG” signalling pathway has been implicated in cell-fate determination, proliferation and the patterning of many different cell types and organs. As the GLI family members (GLI1, GLI2 and GLI3 are key mediators of hedgehog morphogenetic signals, over the past couple of decades they have been extensively scrutinized by genetic, molecular and biochemical means. Thus, a great deal of information is currently available about the functional aspects of GLI proteins in various vertebrate species. To address the roles of GLI genes in diversifying the repertoire of the Hh signalling and deploying them for the vertebrate specifications, in this study we have examined the evolutionary patterns of vertebrate GLI sequences within and between species. Results: Phylogenetic tree analysis suggests that the vertebrate GLI1, GLI2 and GLI3 genes diverged after the separation of urochordates from vertebrates and before the tetrapods-bony fishes split. Lineage specific duplication events were also detected. Estimation of mode and strength of selection acting on GLI orthologs demonstrated that all members of the GLI gene family experienced more relaxed selection in teleost fish than in the mammalian lineage. Furthermore, the GLI1 gene appeared to have been exposed to different functional constraints in fish and tetrapod lineages, whilst a similar level of functional constraints on GLI2 and GLI3 was suggested by comparable average non-synonymous (Ka substitutions across the lineages. A relative rate test suggested that the majority of the paralogous copies of the GLI family analyzed evolved with similar evolutionary rates except GLI1 which evolved at a significantly faster rate than its paralogous counterparts in tetrapods. Conclusions: Our analysis shows that sequence evolutionary patterns of GLI family members are largely correlated with the reported similarities and differences in the functionality of GLI proteins

  19. Adult Gli2+/-;Gli3Δ699/+ Male and Female Mice Display a Spectrum of Genital Malformation.

    Fei He

    Full Text Available Disorders of sexual development (DSD encompass a broad spectrum of urogenital malformations and are amongst the most common congenital birth defects. Although key genetic factors such as the hedgehog (Hh family have been identified, a unifying postnatally viable model displaying the spectrum of male and female urogenital malformations has not yet been reported. Since human cases are diagnosed and treated at various stages postnatally, equivalent mouse models enabling analysis at similar stages are of significant interest. Additionally, all non-Hh based genetic models investigating DSD display normal females, leaving female urogenital development largely unknown. Here, we generated compound mutant mice, Gli2+/-;Gli3Δ699/+, which exhibit a spectrum of urogenital malformations in both males and females upon birth, and also carried them well into adulthood. Analysis of embryonic day (E18.5 and adult mice revealed shortened anogenital distance (AGD, open ventral urethral groove, incomplete fusion of scrotal sac, abnormal penile size and structure, and incomplete testicular descent with hypoplasia in male mice, whereas female mutant mice displayed reduced AGD, urinary incontinence, and a number of uterine anomalies such as vaginal duplication. Male and female fertility was also investigated via breeding cages, and it was identified that male mice were infertile while females were unable to deliver despite becoming impregnated. We propose that Gli2+/-;Gli3Δ699/+ mice can serve as a genetic mouse model for common DSD such as cryptorchidism, hypospadias, and incomplete fusion of the scrotal sac in males, and a spectrum of uterine and vaginal abnormalities along with urinary incontinence in females, which could prove essential in revealing new insights into their equivalent diseases in humans.

  20. Gli3 is a negative regulator of Tas1r3-expressing taste cells

    Jyotaki, Masafumi; Redding, Kevin; Jiang, Peihua

    2018-01-01

    Mouse taste receptor cells survive from 3–24 days, necessitating their regeneration throughout adulthood. In anterior tongue, sonic hedgehog (SHH), released by a subpopulation of basal taste cells, regulates transcription factors Gli2 and Gli3 in stem cells to control taste cell regeneration. Using single-cell RNA-Seq we found that Gli3 is highly expressed in Tas1r3-expressing taste receptor cells and Lgr5+ taste stem cells in posterior tongue. By PCR and immunohistochemistry we found that Gli3 was expressed in taste buds in all taste fields. Conditional knockout mice lacking Gli3 in the posterior tongue (Gli3CKO) had larger taste buds containing more taste cells than did control wild-type (Gli3WT) mice. In comparison to wild-type mice, Gli3CKO mice had more Lgr5+ and Tas1r3+ cells, but fewer type III cells. Similar changes were observed ex vivo in Gli3CKO taste organoids cultured from Lgr5+ taste stem cells. Further, the expression of several taste marker and Gli3 target genes was altered in Gli3CKO mice and/or organoids. Mirroring these changes, Gli3CKO mice had increased lick responses to sweet and umami stimuli, decreased lick responses to bitter and sour taste stimuli, and increased glossopharyngeal taste nerve responses to sweet and bitter compounds. Our results indicate that Gli3 is a suppressor of stem cell proliferation that affects the number and function of mature taste cells, especially Tas1r3+ cells, in adult posterior tongue. Our findings shed light on the role of the Shh pathway in adult taste cell regeneration and may help devise strategies for treating taste distortions from chemotherapy and aging. PMID:29415007

  1. Inflammation and Gli2 suppress gastrin gene expression in a murine model of antral hyperplasia.

    Milena Saqui-Salces

    Full Text Available Chronic inflammation in the stomach can lead to gastric cancer. We previously reported that gastrin-deficient (Gast⁻/⁻ mice develop bacterial overgrowth, inflammatory infiltrate, increased Il-1β expression, antral hyperplasia and eventually antral tumors. Since Hedgehog (Hh signaling is active in gastric cancers but its role in precursor lesions is poorly understood, we examined the role of inflammation and Hh signaling in antral hyperplasia. LacZ reporter mice for Sonic hedgehog (Shh, Gli1, and Gli2 expression bred onto the Gast⁻/⁻ background revealed reduced Shh and Gli1 expression in the antra compared to wild type controls (WT. Gli2 expression in the Gast⁻/⁻ corpus was unchanged. However in the hyperplastic Gast⁻/⁻ antra, Gli2 expression increased in both the mesenchyme and epithelium, whereas expression in WT mice remained exclusively mesenchymal. These observations suggested that Gli2 is differentially regulated in the hyperplastic Gast⁻/⁻ antrum versus the corpus and by a Shh ligand-independent mechanism. Moreover, the proinflammatory cytokines Il-1β and Il-11, which promote gastric epithelial proliferation, were increased in the Gast⁻/⁻ stomach along with Infγ. To test if inflammation could account for elevated epithelial Gli2 expression in the Gast⁻/⁻ antra, the human gastric cell line AGS was treated with IL-1β and was found to increase GLI2 but decrease GLI1 levels. IL-1β also repressed human GAST gene expression. Indeed, GLI2 but not GLI1 or GLI3 expression repressed gastrin luciferase reporter activity by ∼50 percent. Moreover, chromatin immunoprecipitation of GLI2 in AGS cells confirmed that GLI2 directly binds to the GAST promoter. Using a mouse model of constitutively active epithelial GLI2 expression, we found that activated GLI2 repressed Gast expression but induced Il-1β gene expression and proliferation in the gastric antrum, along with a reduction of the number of G-cells. In summary

  2. Targeting GLI by GANT61 involves mechanisms dependent on inhibition of both transcription and DNA licensing.

    Zhang, Ruowen; Wu, Jiahui; Ferrandon, Sylvain; Glowacki, Katie J; Houghton, Janet A

    2016-12-06

    The GLI genes are transcription factors and in cancers are oncogenes, aberrantly and constitutively activated. GANT61, a specific GLI inhibitor, has induced extensive cytotoxicity in human models of colon cancer. The FOXM1 promoter was determined to be a transcriptional target of GLI1. In HT29 cells, inhibition of GLI1 binding at the GLI consensus sequence by GANT61 led to inhibited binding of Pol II, the pause-release factors DSIF, NELF and p-TEFb. The formation of R-loops (RNA:DNA hybrids, ssDNA), were reduced by GANT61 at the FOXM1 promoter. Pretreatment of HT29 cells with α-amanitin reduced GANT61-induced γH2AX foci. Co-localization of GLI1 and BrdU foci, inhibited by GANT61, indicated GLI1 and DNA replication to be linked. By co-immunoprecipitation and confocal microscopy, GLI1 co-localized with the DNA licensing factors ORC4, CDT1, and MCM2. Significant co-localization of GLI1 and ORC4 was inhibited by GANT61, and enrichment of ORC4 occurred at the GLI binding site in the FOXM1 promoter. CDT1 was found to be a transcription target of GLI1. Overexpression of CDT1 in HT29 and SW480 cells reduced GANT61-induced cell death, gH2AX foci, and cleavage of caspase-3. Data demonstrate involvement of transcription and of DNA replication licensing factors by non-transcriptional and transcriptional mechanisms in the GLI-dependent mechanism of action of GANT61.

  3. Peptide analysis as amino alcohols by gas chromatography-mass spectrometry. Application to hyperoligopeptiduria. Detection of Gly-3Hyp-4Hyp and Gly-Pro-4Hyp-Gly.

    Steiner, W; Niederwieser, A

    1979-03-15

    A method for the qualitative analysis of oligopeptides in human urine in cases of peptiduria is described. After sample precleaning on a strongly acidic ion exchanger, the trifluoroacetyl/methyl esters were formed and the peptide derivatives were transformed into trifluoroethyl oligoamino alcohols according to Nau and Biemann. It was found that oligoamino alcohols could be isolated selectively on a weakly acidic ion exchanger. The O-trimethylsilylated trifluoroethyl oligoamino alcohols were separated on a SE-30 glass capillary column and analyzed by computer-assisted gas chromatography-mass spectrometry. In order to increase specificity and to facilitate mass spectrometric interpretation, aliquots of the sample were reduced separately with lithium-aluminium deuteride and hydride. Each peptide gave a pair of derivatives with characteristic mass differences of the ions, namely 2 mass units per reduced oxo group (deuterium-hydrogen-labelling of oxo groups by reduction). Correct identification is assumed only if both mass spectral patterns fit the theory. Sample volumes of 5--100 ml of urine are needed. About six samples can be derivatized per week. Three cases with suspected peptiduria were investigated and the following peptides were found: Gly-Pro-4Hyp-Gly; Gly-Pro-4Hyp; Gly-Hyp-Hyp (postulated isomer Gly-3Hyp-4Hyp); Pro-4Hyp and Gly-Pro. With exception of the tetrapeptide, these compounds could be detected also in the urine of a healthy child.

  4. Pressure dependence of backbone chemical shifts in the model peptides Ac-Gly-Gly-Xxx-Ala-NH2.

    Erlach, Markus Beck; Koehler, Joerg; Crusca, Edson; Kremer, Werner; Munte, Claudia E; Kalbitzer, Hans Robert

    2016-06-01

    For a better understanding of nuclear magnetic resonance (NMR) detected pressure responses of folded as well as unstructured proteins the availability of data from well-defined model systems are indispensable. In this work we report the pressure dependence of chemical shifts of the backbone atoms (1)H(α), (13)C(α) and (13)C' in the protected tetrapeptides Ac-Gly-Gly-Xxx-Ala-NH2 (Xxx one of the 20 canonical amino acids). Contrary to expectation the chemical shifts of these nuclei have a nonlinear dependence on pressure in the range from 0.1 to 200 MPa. The polynomial pressure coefficients B 1 and B 2 are dependent on the type of amino acid studied. The coefficients of a given nucleus show significant linear correlations suggesting that the NMR observable pressure effects in the different amino acids have at least partly the same physical cause. In line with this observation the magnitude of the second order coefficients of nuclei being direct neighbors in the chemical structure are also weakly correlated.

  5. Pressure dependence of backbone chemical shifts in the model peptides Ac-Gly-Gly-Xxx-Ala-NH{sub 2}

    Erlach, Markus Beck; Koehler, Joerg [University of Regensburg, Institute of Biophysics and Physical Biochemistry and Centre of Magnetic Resonance in Chemistry and Biomedicine (Germany); Crusca, Edson [University of São Paulo, Physics Institute of São Carlos (Brazil); Kremer, Werner [University of Regensburg, Institute of Biophysics and Physical Biochemistry and Centre of Magnetic Resonance in Chemistry and Biomedicine (Germany); Munte, Claudia E. [University of São Paulo, Physics Institute of São Carlos (Brazil); Kalbitzer, Hans Robert, E-mail: hans-robert.kalbitzer@biologie.uni-regensburg.de [University of Regensburg, Institute of Biophysics and Physical Biochemistry and Centre of Magnetic Resonance in Chemistry and Biomedicine (Germany)

    2016-06-15

    For a better understanding of nuclear magnetic resonance (NMR) detected pressure responses of folded as well as unstructured proteins the availability of data from well-defined model systems are indispensable. In this work we report the pressure dependence of chemical shifts of the backbone atoms {sup 1}H{sup α}, {sup 13}C{sup α} and {sup 13}C′ in the protected tetrapeptides Ac-Gly-Gly-Xxx-Ala-NH{sub 2} (Xxx one of the 20 canonical amino acids). Contrary to expectation the chemical shifts of these nuclei have a nonlinear dependence on pressure in the range from 0.1 to 200 MPa. The polynomial pressure coefficients B{sub 1} and B{sub 2} are dependent on the type of amino acid studied. The coefficients of a given nucleus show significant linear correlations suggesting that the NMR observable pressure effects in the different amino acids have at least partly the same physical cause. In line with this observation the magnitude of the second order coefficients of nuclei being direct neighbors in the chemical structure are also weakly correlated.Graphical Abstract.

  6. Application of evolutionary algorithm methods to polypeptide folding: comparison with experimental results for unsolvated Ac-(Ala-Gly-Gly)5-LysH+

    Damsbo, Martin; Kinnear, Brian S; Hartings, Matthew R

    2004-01-01

    We present an evolutionary method for finding the low-energy conformations of polypeptides. The application, called FOLDAWAY,is based on a generic framework and uses several evolutionary operators as well as local optimization to navigate the complex energy landscape of polypeptides. It maintains...... mobility measurements. It has a flat energy landscape where helical and globular conformations have similar energies. FOLDAWAY locates several large groups of structures not found in previous molecular dynamics simulations for this peptide, including compact globular conformations, which are probably...... two complementary representations of the structures and uses the CHARMM force field for evaluating the energies. The method is applied to unsolvated Met-enkephalin and Ac-(Ala-Gly-Gly)(5)-Lys(+)H(+). Unsolvated Ac-(Ala-Gly-Gly)(5)-Lys(+)H(+) has been the object of recent experimental studies using ion...

  7. Effects of Pro-Gly-Pro tripeptide on the dopamine system.

    Meshavkin, V K; Batishcheva, E Yu; Kost, N V; Sokolov, O Yu; Trufanova, A V; Samonina, G E

    2011-08-01

    Tripeptide Pro-Gly-Pro interacted with dopamine receptors in vitro and reduced behavioral manifestations of apomorphine-induced hyperfunction of the dopamine system in verticalization, stereotypy, and yawning tests. Presumably, the behavioral effects of Pro-Gly-Pro tripeptide were mediated through post- and presynaptic D(2)and D(3)receptors.

  8. Pressure dependence of side chain 13C chemical shifts in model peptides Ac-Gly-Gly-Xxx-Ala-NH2.

    Beck Erlach, Markus; Koehler, Joerg; Crusca, Edson; Munte, Claudia E; Kainosho, Masatsune; Kremer, Werner; Kalbitzer, Hans Robert

    2017-10-01

    For evaluating the pressure responses of folded as well as intrinsically unfolded proteins detectable by NMR spectroscopy the availability of data from well-defined model systems is indispensable. In this work we report the pressure dependence of 13 C chemical shifts of the side chain atoms in the protected tetrapeptides Ac-Gly-Gly-Xxx-Ala-NH 2 (Xxx, one of the 20 canonical amino acids). Contrary to expectation the chemical shifts of a number of nuclei have a nonlinear dependence on pressure in the range from 0.1 to 200 MPa. The size of the polynomial pressure coefficients B 1 and B 2 is dependent on the type of atom and amino acid studied. For H N , N and C α the first order pressure coefficient B 1 is also correlated to the chemical shift at atmospheric pressure. The first and second order pressure coefficients of a given type of carbon atom show significant linear correlations suggesting that the NMR observable pressure effects in the different amino acids have at least partly the same physical cause. In line with this observation the magnitude of the second order coefficients of nuclei being direct neighbors in the chemical structure also are weakly correlated. The downfield shifts of the methyl resonances suggest that gauche conformers of the side chains are not preferred with pressure. The valine and leucine methyl groups in the model peptides were assigned using stereospecifically 13 C enriched amino acids with the pro-R carbons downfield shifted relative to the pro-S carbons.

  9. Gli3 Regulation of Myogenesis Is Necessary for Ischemia-Induced Angiogenesis

    Renault, Marie-Ange; Vandierdonck, Soizic; Chapouly, Candice; Yu, Yang; Qin, Gangjian; Metras, Alexandre; Couffinhal, Thierry; Losordo, Douglas W.; Yao, Qinyu; Reynaud, Annabel; Jaspard-Vinassa, Béatrice; Belloc, Isabelle; Desgranges, Claude; Gadeau, Alain-Pierre

    2015-01-01

    Rationale A better understanding of the mechanism underlying skeletal muscle repair is required to develop therapies that promote tissue regeneration in adults. Hedgehog signaling has been shown previously to be involved in myogenesis and angiogenesis: 2 crucial processes for muscle development and regeneration. Objective The objective of this study was to identify the role of the hedgehog transcription factor Gli3 in the crosstalk between angiogenesis and myogenesis in adults. Methods and Results Using conditional knockout mice, we found that Gli3 deficiency in endothelial cells did not affect ischemic muscle repair, whereas in myocytes, Gli3 deficiency resulted in severely delayed ischemia-induced myogenesis. Moreover, angiogenesis was also significantly impaired in HSA-CreERT2; Gli3Flox/Flox mice, demonstrating that impaired myogenesis indirectly affects ischemia-induced angiogenesis. The role of Gli3 in myocytes was then further investigated. We found that Gli3 promotes myoblast differentiation through myogenic factor 5 regulation. In addition, we found that Gli3 regulates several proangiogenic factors, including thymidine phosphorylase and angiopoietin-1 both in vitro and in vivo, which indirectly promote endothelial cell proliferation and arteriole formation. In addition, we found that Gli3 is upregulated in proliferating myoblasts by the cell cycle–associated transcription factor E2F1. Conclusions This study shows for the first time that Gli3-regulated postnatal myogenesis is necessary for muscle repair–associated angiogenesis. Most importantly, it implies that myogenesis drives angiogenesis in the setting of skeletal muscle repair and identifies Gli3 as a potential target for regenerative medicine. PMID:24044950

  10. Identification of RegIV as a novel GLI1 target gene in human pancreatic cancer.

    Feng Wang

    2011-04-01

    Full Text Available GLI1 is the key transcriptional factor in the Hedgehog signaling pathway in pancreatic cancer. RegIV is associated with regeneration, and cell growth, survival, adhesion and resistance to apoptosis. We aimed to study RegIV expression in pancreatic cancer and its relationship to GLI1.GLI1 and RegIV expression were evaluated in tumor tissue and adjacent normal tissues of pancreatic cancer patients and 5 pancreatic cancer cell lines by qRT-PCR, Western blot, and immunohistochemistry (IHC, and the correlation between them. The GLI1-shRNA lentiviral vector was constructed and transfected into PANC-1, and lentiviral vector containing the GLI1 expression sequence was constructed and transfected into BxPC-3. GLI1 and RegIV expression were evaluated by qRT-PCR and Western blot. Finally we demonstrated RegIV to be the target of GLI1 by chromatin immunoprecipitation (CHIP and electrophoretic mobility shift assays (EMSA.The results of IHC and qRT-PCR showed that RegIV and GLI1 expression was higher in pancreatic cancer tissues versus adjacent normal tissues (p<0.001. RegIV expression correlated with GLI1 expression in these tissues (R = 0.795, p<0.0001. These results were verified for protein (R = 0.939, p = 0.018 and mRNA expression (R = 0.959, p = 0.011 in 5 pancreatic cancer cell lines. RegIV mRNA and protein expression was decreased (94.7±0.3%, 84.1±0.5%; respectively when GLI1 was knocked down (82.1±3.2%, 76.7±2.2%; respectively by the RNAi technique. GLI1 overexpression in mRNA and protein level (924.5±5.3%, 362.1±3.5%; respectively induced RegIV overexpression (729.1±4.3%, 339.0±3.7%; respectively. Moreover, CHIP and EMSA assays showed GLI1 protein bound to RegIV promotor regions (GATCATCCA in pancreatic cancer cells.GLI1 promotes RegIV transcription by binding to the RegIV gene promoter in pancreatic cancer.

  11. A mechanism for vertebrate Hedgehog signaling: recruitment to cilia and dissociation of SuFu–Gli protein complexes

    Tukachinsky, Hanna; Lopez, Lyle V.; Salic, Adrian

    2010-01-01

    In vertebrates, Hedgehog (Hh) signaling initiated in primary cilia activates the membrane protein Smoothened (Smo) and leads to activation of Gli proteins, the transcriptional effectors of the pathway. In the absence of signaling, Gli proteins are inhibited by the cytoplasmic protein Suppressor of Fused (SuFu). It is unclear how Hh activates Gli and whether it directly regulates SuFu. We find that Hh stimulation quickly recruits endogenous SuFu–Gli complexes to cilia, suggesting a model in wh...

  12. Penetrance of eye defects in mice heterozygous for mutation of Gli3 is enhanced by heterozygous mutation of Pax6

    Price David J

    2006-10-01

    Full Text Available Abstract Background Knowledge of the consequences of heterozygous mutations of developmentally important genes is important for understanding human genetic disorders. The Gli3 gene encodes a zinc finger transcription factor and homozygous loss-of-function mutations of Gli3 are lethal. Humans heterozygous for mutations in this gene suffer Greig cephalopolysyndactyly or Pallister-Hall syndromes, in which limb defects are prominent, and mice heterozygous for similar mutations have extra digits. Here we examined whether eye development, which is abnormal in mice lacking functional Gli3, is defective in Gli3+/- mice. Results We showed that Gli3 is expressed in the developing eye but that Gli3+/- mice have only very subtle eye defects. We then generated mice compound heterozygous for mutations in both Gli3 and Pax6, which encodes another developmentally important transcription factor known to be crucial for eye development. Pax6+/-; Gli3+/- eyes were compared to the eyes of wild-type, Pax6+/- or Gli3+/- siblings. They exhibited a range of abnormalities of the retina, iris, lens and cornea that was more extensive than in single Gli3+/- or Pax6+/- mutants or than would be predicted by addition of their phenotypes. Conclusion These findings indicate that heterozygous mutations of Gli3 can impact on eye development. The importance of a normal Gli3 gene dosage becomes greater in the absence of a normal Pax6 gene dosage, suggesting that the two genes co-operate during eye morphogenesis.

  13. Ihh/Gli2 signaling promotes osteoblast differentiation by regulating Runx2 expression and function.

    Shimoyama, Atsuko; Wada, Masahiro; Ikeda, Fumiyo; Hata, Kenji; Matsubara, Takuma; Nifuji, Akira; Noda, Masaki; Amano, Katsuhiko; Yamaguchi, Akira; Nishimura, Riko; Yoneda, Toshiyuki

    2007-07-01

    Genetic and cell biological studies have indicated that Indian hedgehog (Ihh) plays an important role in bone development and osteoblast differentiation. However, the molecular mechanism by which Ihh regulates osteoblast differentiation is complex and remains to be fully elucidated. In this study, we investigated the role of Ihh signaling in osteoblast differentiation using mesenchymal cells and primary osteoblasts. We observed that Ihh stimulated alkaline phosphatase (ALP) activity, osteocalcin expression, and calcification. Overexpression of Gli2- but not Gli3-induced ALP, osteocalcin expression, and calcification of these cells. In contrast, dominant-negative Gli2 markedly inhibited Ihh-dependent osteoblast differentiation. Ihh treatment or Gli2 overexpression also up-regulated the expression of Runx2, an essential transcription factor for osteoblastogenesis, and enhanced the transcriptional activity and osteogenic action of Runx2. Coimmunoprecipitation analysis demonstrated a physical interaction between Gli2 and Runx2. Moreover, Ihh or Gli2 overexpression failed to increase ALP activity in Runx2-deficient mesenchymal cells. Collectively, these results suggest that Ihh regulates osteoblast differentiation of mesenchymal cells through up-regulation of the expression and function of Runx2 by Gli2.

  14. New insights into genotype-phenotype correlation for GLI3 mutations.

    Démurger, Florence; Ichkou, Amale; Mougou-Zerelli, Soumaya; Le Merrer, Martine; Goudefroye, Géraldine; Delezoide, Anne-Lise; Quélin, Chloé; Manouvrier, Sylvie; Baujat, Geneviève; Fradin, Mélanie; Pasquier, Laurent; Megarbané, André; Faivre, Laurence; Baumann, Clarisse; Nampoothiri, Sheela; Roume, Joëlle; Isidor, Bertrand; Lacombe, Didier; Delrue, Marie-Ange; Mercier, Sandra; Philip, Nicole; Schaefer, Elise; Holder, Muriel; Krause, Amanda; Laffargue, Fanny; Sinico, Martine; Amram, Daniel; André, Gwenaelle; Liquier, Alain; Rossi, Massimiliano; Amiel, Jeanne; Giuliano, Fabienne; Boute, Odile; Dieux-Coeslier, Anne; Jacquemont, Marie-Line; Afenjar, Alexandra; Van Maldergem, Lionel; Lackmy-Port-Lis, Marylin; Vincent-Delorme, Catherine; Chauvet, Marie-Liesse; Cormier-Daire, Valérie; Devisme, Louise; Geneviève, David; Munnich, Arnold; Viot, Géraldine; Raoul, Odile; Romana, Serge; Gonzales, Marie; Encha-Razavi, Ferechte; Odent, Sylvie; Vekemans, Michel; Attie-Bitach, Tania

    2015-01-01

    The phenotypic spectrum of GLI3 mutations includes autosomal dominant Greig cephalopolysyndactyly syndrome (GCPS) and Pallister-Hall syndrome (PHS). PHS was first described as a lethal condition associating hypothalamic hamartoma, postaxial or central polydactyly, anal atresia and bifid epiglottis. Typical GCPS combines polysyndactyly of hands and feet and craniofacial features. Genotype-phenotype correlations have been found both for the location and the nature of GLI3 mutations, highlighting the bifunctional nature of GLI3 during development. Here we report on the molecular and clinical study of 76 cases from 55 families with either a GLI3 mutation (49 GCPS and 21 PHS), or a large deletion encompassing the GLI3 gene (6 GCPS cases). Most of mutations are novel and consistent with the previously reported genotype-phenotype correlation. Our results also show a correlation between the location of the mutation and abnormal corpus callosum observed in some patients with GCPS. Fetal PHS observations emphasize on the possible lethality of GLI3 mutations and extend the phenotypic spectrum of malformations such as agnathia and reductional limbs defects. GLI3 expression studied by in situ hybridization during human development confirms its early expression in target tissues.

  15. New insights into genotype–phenotype correlation for GLI3 mutations

    Démurger, Florence; Ichkou, Amale; Mougou-Zerelli, Soumaya; Le Merrer, Martine; Goudefroye, Géraldine; Delezoide, Anne-Lise; Quélin, Chloé; Manouvrier, Sylvie; Baujat, Geneviève; Fradin, Mélanie; Pasquier, Laurent; Megarbané, André; Faivre, Laurence; Baumann, Clarisse; Nampoothiri, Sheela; Roume, Joëlle; Isidor, Bertrand; Lacombe, Didier; Delrue, Marie-Ange; Mercier, Sandra; Philip, Nicole; Schaefer, Elise; Holder, Muriel; Krause, Amanda; Laffargue, Fanny; Sinico, Martine; Amram, Daniel; André, Gwenaelle; Liquier, Alain; Rossi, Massimiliano; Amiel, Jeanne; Giuliano, Fabienne; Boute, Odile; Dieux-Coeslier, Anne; Jacquemont, Marie-Line; Afenjar, Alexandra; Van Maldergem, Lionel; Lackmy-Port-Lis, Marylin; Vincent- Delorme, Catherine; Chauvet, Marie-Liesse; Cormier-Daire, Valérie; Devisme, Louise; Geneviève, David; Munnich, Arnold; Viot, Géraldine; Raoul, Odile; Romana, Serge; Gonzales, Marie; Encha-Razavi, Ferechte; Odent, Sylvie; Vekemans, Michel; Attie-Bitach, Tania

    2015-01-01

    The phenotypic spectrum of GLI3 mutations includes autosomal dominant Greig cephalopolysyndactyly syndrome (GCPS) and Pallister–Hall syndrome (PHS). PHS was first described as a lethal condition associating hypothalamic hamartoma, postaxial or central polydactyly, anal atresia and bifid epiglottis. Typical GCPS combines polysyndactyly of hands and feet and craniofacial features. Genotype–phenotype correlations have been found both for the location and the nature of GLI3 mutations, highlighting the bifunctional nature of GLI3 during development. Here we report on the molecular and clinical study of 76 cases from 55 families with either a GLI3 mutation (49 GCPS and 21 PHS), or a large deletion encompassing the GLI3 gene (6 GCPS cases). Most of mutations are novel and consistent with the previously reported genotype–phenotype correlation. Our results also show a correlation between the location of the mutation and abnormal corpus callosum observed in some patients with GCPS. Fetal PHS observations emphasize on the possible lethality of GLI3 mutations and extend the phenotypic spectrum of malformations such as agnathia and reductional limbs defects. GLI3 expression studied by in situ hybridization during human development confirms its early expression in target tissues. PMID:24736735

  16. Brasile 1960: Gli anni della svolta per Alberto Moravia

    Simone Casini

    2016-06-01

    Full Text Available Alberto Moravia arrived in Brazil in the summer of 1960 to preside the Congress of the PEN Club. He had recently finished La noia, his eleventh novel, the one that most directly is connected to his first novel, Gli indifferenti (1929, and it marks a decisive turning point in his artistic, intellectual, and human course. The essay reconstructs the life and work of the 50-year-old writer, in which a 10-year research is summarized and concluded, opening a new phase. Rereading the articles written for Il Corriere della Sera and the review of the Quarto de despejo by Carolina de Jesus, we focused on the origin and acute reflection of Moravia about Brazil in those years (Brasilia, Bahia, Rio, between past and future. The experience of Brazil, for the writer, who in those following years will attempt new paths in the intellectual and literary field, and new goals for his travels, always more oriented, in the company of Dacia Maraini and Pier Paolo Pasolini, toward the Third World.

  17. Signal interaction of Hedgehog/GLI and epidermal growth factor receptor signaling in cancer development

    Eberl, M.

    2012-01-01

    The subject of this PhD thesis is based on the cooperation of Hedgehog (HH)/GLI with epidermal growth factor receptor (EGFR) signaling synergistically promoting oncogenic transformation and cancer growth. In previous studies we have demonstrated that the HH/GLI and EGFR signaling pathways interact synergistically resulting not only in selective induction of HH/GLI-EGFR target genes, but also in the onset of oncogenic transformation and tumor formation (Kasper, Schnidar et al. 2006; Schnidar, Eberl et al. 2009). However, the molecular key mediators acting downstream of HH/GLI and EGFR signal cooperation were largely unknown and the in vivo evidence for the therapeutic relevance of HH/GLI and EGFR signal cooperation in HH-associated cancers was lacking. During my PhD thesis I could demonstrate that the integration of EGFR and HH/GLI signaling involves activation of RAS/MEK/ERK and JUN/AP1 signaling in response to EGFR activation. Furthermore I succeeded in identifying genes, including stem cell- (SOX2, SOX9), tumor growth- (JUN, TGFA, FGF19) and metastasis-associated genes (SPP1/osteopontin, CXCR4) that showed synergistic transcriptional activation by HH/GLI-EGFR signal integration. Importantly, I could demonstrate that these genes arrange themselves within a stable interdependent signaling network, which is required for in vivo growth of basal cell carcinoma (BCC) and tumor-initiating pancreatic cancer cells. These data validate EGFR signaling as additional drug target in HH/GLI driven cancers and provide new therapeutic strategies based on combined targeting of cooperative HH/GLI-EGFR signaling and selected downstream target genes (Eberl, Klingler et al. 2012). (author) [de

  18. The Gli2 transcriptional activator is a crucial effector for Ihh signaling in osteoblast development and cartilage vascularization.

    Joeng, Kyu Sang; Long, Fanxin

    2009-12-01

    Indian hedgehog (Ihh) critically regulates multiple aspects of endochondral bone development. Although it is generally believed that all Ihh functions are mediated by the Gli family of transcription activators and repressors, formal genetic proof for this notion has not been provided. Moreover, the extent to which different Gli proteins contribute to Ihh functions is not fully understood. Previous work has shown that de-repression of the Gli3 repressor is the predominant mode through which Ihh controls chondrocyte proliferation and maturation, but that osteoblast differentiation and hypertrophic cartilage vascularization require additional mechanisms. To test the involvement of Gli2 activation in these processes, we have generated a mouse strain that expresses a constitutive Gli2 activator in a Cre-dependent manner, and have attempted to rescue the Ihh-null mouse with the Gli2 activator, either alone or in combination with Gli3 removal. Here, we report that the Gli2 activator alone is sufficient to induce vascularization of the hypertrophic cartilage in the absence of Ihh but requires simultaneous removal of Gli3 to restore osteoblast differentiation. These results therefore provide direct genetic evidence that Gli2 and Gli3 collectively mediate all major aspects of Ihh function during endochondral skeletal development.

  19. Histone acetyltransferase PCAF is required for Hedgehog-Gli-dependent transcription and cancer cell proliferation

    Malatesta, Martina; Steinhauer, Cornelia; Mohammad, Faizaan

    2013-01-01

    The Hedgehog (Hh) signaling pathway plays an important role in embryonic patterning and development of many tissues and organs as well as in maintaining and repairing mature tissues in adults. Uncontrolled activation of the Hh-Gli pathway has been implicated in developmental abnormalities as well...... that the histone acetyltransferase PCAF/KAT2B is an important factor of the Hh pathway. Specifically, we show that PCAF depletion impairs Hh activity and reduces expression of Hh target genes. Consequently, PCAF downregulation in medulloblastoma and glioblastoma cells leads to decreased proliferation and increased...... apoptosis. In addition, we found that PCAF interacts with GLI1, the downstream effector in the Hh-Gli pathway, and that PCAF or GLI1 loss reduces the levels of H3K9 acetylation on Hh target gene promoters. Finally, we observed that PCAF silencing reduces the tumor-forming potential of neural stem cells...

  20. Synthesis and Characterization of Novel Acyl-Glycine Inhibitors of GlyT2.

    Mostyn, Shannon N; Carland, Jane E; Shimmon, Susan; Ryan, Renae M; Rawling, Tristan; Vandenberg, Robert J

    2017-09-20

    It has been demonstrated previously that the endogenous compound N-arachidonyl-glycine inhibits the glycine transporter GlyT2, stimulates glycinergic neurotransmission, and provides analgesia in animal models of neuropathic and inflammatory pain. However, it is a relatively weak inhibitor with an IC 50 of 9 μM and is subject to oxidation via cyclooxygenase, limiting its therapeutic value. In this paper we describe the synthesis and testing of a novel series of monounsaturated C18 and C16 acyl-glycine molecules as inhibitors of the glycine transporter GlyT2. We demonstrate that they are up to 28 fold more potent that N-arachidonyl-glycine with no activity at the closely related GlyT1 transporter at concentrations up to 30 μM. This novel class of compounds show considerable promise as a first generation of GlyT2 transport inhibitors.

  1. Identification of a 3rd Na+ Binding Site of the Glycine Transporter, GlyT2.

    Nandhitha Subramanian

    Full Text Available The Na+/Cl- dependent glycine transporters GlyT1 and GlyT2 regulate synaptic glycine concentrations. Glycine transport by GlyT2 is coupled to the co-transport of three Na+ ions, whereas transport by GlyT1 is coupled to the co-transport of only two Na+ ions. These differences in ion-flux coupling determine their respective concentrating capacities and have a direct bearing on their functional roles in synaptic transmission. The crystal structures of the closely related bacterial Na+-dependent leucine transporter, LeuTAa, and the Drosophila dopamine transporter, dDAT, have allowed prediction of two Na+ binding sites in GlyT2, but the physical location of the third Na+ site in GlyT2 is unknown. A bacterial betaine transporter, BetP, has also been crystallized and shows structural similarity to LeuTAa. Although betaine transport by BetP is coupled to the co-transport of two Na+ ions, the first Na+ site is not conserved between BetP and LeuTAa, the so called Na1' site. We hypothesized that the third Na+ binding site (Na3 site of GlyT2 corresponds to the BetP Na1' binding site. To identify the Na3 binding site of GlyT2, we performed molecular dynamics (MD simulations. Surprisingly, a Na+ placed at the location consistent with the Na1' site of BetP spontaneously dissociated from its initial location and bound instead to a novel Na3 site. Using a combination of MD simulations of a comparative model of GlyT2 together with an analysis of the functional properties of wild type and mutant GlyTs we have identified an electrostatically favorable novel third Na+ binding site in GlyT2 formed by Trp263 and Met276 in TM3, Ala481 in TM6 and Glu648 in TM10.

  2. New insights into genotype–phenotype correlation for GLI3 mutations

    Démurger, Florence; Ichkou, Amale; Mougou-Zerelli, Soumaya; Le Merrer, Martine; Goudefroye, Géraldine; Delezoide, Anne-Lise; Quélin, Chloé; Manouvrier, Sylvie; Baujat, Geneviève; Fradin, Mélanie; Pasquier, Laurent; Megarbané, André; Faivre, Laurence; Baumann, Clarisse; Nampoothiri, Sheela

    2014-01-01

    The phenotypic spectrum of GLI3 mutations includes autosomal dominant Greig cephalopolysyndactyly syndrome (GCPS) and Pallister–Hall syndrome (PHS). PHS was first described as a lethal condition associating hypothalamic hamartoma, postaxial or central polydactyly, anal atresia and bifid epiglottis. Typical GCPS combines polysyndactyly of hands and feet and craniofacial features. Genotype–phenotype correlations have been found both for the location and the nature of GLI3 mutations, highlightin...

  3. A genome scale RNAi screen identifies GLI1 as a novel gene regulating vorinostat sensitivity.

    Falkenberg, K J; Newbold, A; Gould, C M; Luu, J; Trapani, J A; Matthews, G M; Simpson, K J; Johnstone, R W

    2016-07-01

    Vorinostat is an FDA-approved histone deacetylase inhibitor (HDACi) that has proven clinical success in some patients; however, it remains unclear why certain patients remain unresponsive to this agent and other HDACis. Constitutive STAT (signal transducer and activator of transcription) activation, overexpression of prosurvival Bcl-2 proteins and loss of HR23B have been identified as potential biomarkers of HDACi resistance; however, none have yet been used to aid the clinical utility of HDACi. Herein, we aimed to further elucidate vorinostat-resistance mechanisms through a functional genomics screen to identify novel genes that when knocked down by RNA interference (RNAi) sensitized cells to vorinostat-induced apoptosis. A synthetic lethal functional screen using a whole-genome protein-coding RNAi library was used to identify genes that when knocked down cooperated with vorinostat to induce tumor cell apoptosis in otherwise resistant cells. Through iterative screening, we identified 10 vorinostat-resistance candidate genes that sensitized specifically to vorinostat. One of these vorinostat-resistance genes was GLI1, an oncogene not previously known to regulate the activity of HDACi. Treatment of vorinostat-resistant cells with the GLI1 small-molecule inhibitor, GANT61, phenocopied the effect of GLI1 knockdown. The mechanism by which GLI1 loss of function sensitized tumor cells to vorinostat-induced apoptosis is at least in part through interactions with vorinostat to alter gene expression in a manner that favored apoptosis. Upon GLI1 knockdown and vorinostat treatment, BCL2L1 expression was repressed and overexpression of BCL2L1 inhibited GLI1-knockdown-mediated vorinostat sensitization. Taken together, we present the identification and characterization of GLI1 as a new HDACi resistance gene, providing a strong rationale for development of GLI1 inhibitors for clinical use in combination with HDACi therapy.

  4. GLI1 interferes with the DNA mismatch repair system in pancreatic cancer through BHLHE41-mediated suppression of MLH1.

    Inaguma, Shingo; Riku, Miho; Hashimoto, Mitsuyoshi; Murakami, Hideki; Saga, Shinsuke; Ikeda, Hiroshi; Kasai, Kenji

    2013-12-15

    The mismatch repair (MMR) system is indispensable for the fidelity of DNA replication, the impairment of which predisposes to the development and progression of many types of cancers. To date, GLI1 transcription factor, a key molecule of the Hedgehog signaling pathway, has been shown to regulate the expression of several genes crucial for a variety of cancer cell properties in many types of cancers, including pancreatic ductal adenocarcinoma (PDAC), but whether GLI1 could control the MMR system was not known. Here, we showed that GLI1 and GLI2 indirectly suppressed the expression of MLH1 in PDAC cells. Through GLI1 target gene screening, we found that GLI1 and GLI2 activated the expression of a basic helix-loop-helix type suppressor BHLHE41/DEC2/SHARP1 through a GLI-binding site in the promoter. Consistent with a previous report that BHLHE41 suppresses the MLH1 promoter activity, we found that the activation of GLI1 led to the BHLHE41-dependent suppression of MLH1, and a double knockdown of GLI1 and GLI2 conversely increased the MLH1 protein in PDAC cells. Using TALEN-based modification of the MLH1 gene, we further showed that GLI1 expression was indeed associated with an increased tolerance to a methylating agent, methylnitrosourea cooperatively with a lower copy number status of MLH1. Finally, GLI1 expression was immunohistochemically related positively with BHLHE41 and inversely with MLH1 in PDAC cells and precancerous lesions of the pancreas. On the basis of these results, we propose that GLI1 depresses the MMR activity and might contribute to the development and progression of PDAC. ©2013 AACR.

  5. The transcription factor Foxc1 is necessary for Ihh-Gli2-regulated endochondral ossification.

    Yoshida, Michiko; Hata, Kenji; Takashima, Rikako; Ono, Koichiro; Nakamura, Eriko; Takahata, Yoshifumi; Murakami, Tomohiko; Iseki, Sachiko; Takano-Yamamoto, Teruko; Nishimura, Riko; Yoneda, Toshiyuki

    2015-03-26

    Indian hedgehog (Ihh) regulates endochondral ossification in both a parathyroid hormone-related protein (PTHrP)-dependent and -independent manner by activating transcriptional mediator Gli2. However, the molecular mechanisms underlying these processes remain elusive. Here by using in vivo microarray analysis, we identify forkhead box C1 (Foxc1) as a transcriptional partner of Gli2. Foxc1 stimulates expression of Ihh target genes, including PTHrP and Col10a1, through its physical and functional interaction with Gli2. Conversely, a dominant negative Foxc1 inhibits the Ihh target gene expression. In a spontaneous loss of Foxc1 function mouse (Foxc1(ch/ch)), endochondral ossification is delayed and the expression of Ihh target genes inhibited. Moreover, the pathological Foxc1 missense mutation observed in the Axenfeld-Rieger syndrome impairs Gli2-Foxc1 association as well as Ihh function. Our findings suggest that Foxc1 is an important transcriptional partner of Ihh-Gli2 signalling during endochondral ossification, and that disruption of the Foxc1-Gli2 interaction causes skeletal abnormalities observed in the Axenfeld-Rieger syndrome.

  6. Association between Toll-like receptor 4 Asp299Gly polymorphism and coronary heart disease susceptibility.

    Wu, B W; Zhu, J; Shi, H M; Jin, B; Wen, Z C

    2017-08-07

    Published data on the association between Toll-like receptor 4 (TLR4) Asp299Gly polymorphism and coronary heart disease (CHD) susceptibility are inconclusive. To derive a more precise estimation of the relationship, a meta-analysis was performed. English-language studies were identified by searching PubMed and Embase databases (up to November 2016). All epidemiological studies were regarding Caucasians because no TLR4 Asp/Gly and Gly/Gly genotypes have been detected in Asians. A total of 20 case-control studies involving 14,416 cases and 10,764 controls were included in the meta-analysis. Overall, no significant associations were found between TLR4 Asp299Gly polymorphism and CHD susceptibility in the dominant model (OR=0.89; 95%CI=0.74 to 1.06; P=0.20) pooled in the meta-analysis. In the subgroup analysis by CHD, non-significant associations were found in cases compared to controls. When stratified by control source, no significantly decreased risk was found in the additive model or dominant model. The present meta-analysis suggests that the TLR4 Asp299Gly polymorphism was not associated with decreased CHD risk in Caucasians.

  7. Gli studi provenzali nel percorso critico di Cesare De Lollis

    Diego Stefanelli

    2015-12-01

    Full Text Available L’intervento indaga l’attività di provenzalista di Cesare De Lollis. Sulla scia dell’opera del maestro Ernesto Monaci essa si concentrò inizialmente sull’edizione di codici provenzali e sullo studio dei primordi della provenzalistica nell’umanesimo italiano. Seguì l’edizione dell’opera di Sordello (1896 che innescò un’aspra polemica con Francesco Torraca, da collocarsi nel contesto della querelle tra critica estetica ed erudita. In seguito, la letteratura dei trovatori fu studiata da De Lollis nel suo rapporto con quella italiana. Ripercorrendo i suoi principali interventi di materia provenzale posteriori all’edizione di Sordello, si dimostra come proprio dal confronto tra le due letterature De Lollis individuò una  linea (che dagli ultimi provenzali arrivava fino a Dante, passando per gli stilnovisti e Guittone caratterizzata dal primato della forma. Tale linea fu fondamentale per l’elaborazione di una delle idee forti della critica dello studioso, la dialettica tra tradizione letteraria codificata e margini di libertà espressiva degli autori in essa operanti, che sarà alla base dei Saggi sulla forma poetica italiana dell’Ottocento.The article focuses on Cesare De Lollis’ studies on Occitan Litterature. At first, following Ernesto Monaci’s example, he worked on the editions of Occitan codes and he studied the beginnings of Occitan philology during the Italian Umanesimo. In 1896 he published the critical edition of Sordello, entering into debate with Francesco Torraca: this debate has to be set in the context of the querelle between critica estetica e critica erudita, that was very strong in Italy at the end of XIX century. After the Sordello’s edition, De Lollis studied the relationship between Occitan and Italian Litterature and he identified a poetic tradition (the last troubadours, Guittone, the stilnovisti, Dante, characterized by the importance of the forma. He focused also on the dialectic between a

  8. GlyT1 Inhibitor NFPS Exerts Neuroprotection via GlyR Alpha1 Subunit in the Rat Model of Transient Focal Cerebral Ischaemia and Reperfusion

    Baosheng Huang

    2016-05-01

    Full Text Available Background/Aims: Glycine is a strychnine-sensitive inhibitory neurotransmitter in the central nervous system (CNS, especially in the spinal cord, brainstem, and retina. The objective of the present study was to investigate the potential neuroprotective effects of GlyT1 inhibitor N [3-(4'-fluorophenyl-3-(4'-phenylphenoxy propyl] sarcosine (NFPS in the rat model of experimental stroke. Methods: In vivo ischaemia was induced by transient middle cerebral artery occlusion (tMCAO. The methods of Western Blotting, Nissl Staining and Morris water maze methods were applied to analyze the anti-ischaemia mechanism. Results: The results showed that high dose of NFPS (H-NFPS significantly reduced infarct volume, neuronal injury and the expression of cleaved caspase-3, enhanced Bcl-2/Bax, and improved spatial learning deficits which were administered three hours after transient middle cerebral artery occlusion (tMCAO induction in rats, while, low dose of NFPS (L-NFPS exacerbated the injury of ischaemia. These findings suggested that low and high dose of NFPS produced opposite effects. Importantly, it was demonstrated that H-NFPS-dependent neuronal protection was inverted by salicylate (Sal, a specific GlyR ɑ1 antagonist. Such effects could probably be attributed to the enhanced glycine level in both synaptic and extrasynaptic clefts and the subsequently altered extrasynaptic GlyRs and their subtypes. Conclusions: These data imply that GlyT1 inhibitor NFPS may be a novel target for clinical treatment of transient focal cerebral ischaemia and reperfusion which are associated with altered GlyR alpha 1 subunits.

  9. Elongation Factor Tu Prevents Misediting of Gly-tRNA(Gly Caused by the Design Behind the Chiral Proofreading Site of D-Aminoacyl-tRNA Deacylase.

    Satya Brata Routh

    2016-05-01

    Full Text Available D-aminoacyl-tRNA deacylase (DTD removes D-amino acids mischarged on tRNAs and is thus implicated in enforcing homochirality in proteins. Previously, we proposed that selective capture of D-aminoacyl-tRNA by DTD's invariant, cross-subunit Gly-cisPro motif forms the mechanistic basis for its enantioselectivity. We now show, using nuclear magnetic resonance (NMR spectroscopy-based binding studies followed by biochemical assays with both bacterial and eukaryotic systems, that DTD effectively misedits Gly-tRNAGly. High-resolution crystal structure reveals that the architecture of DTD's chiral proofreading site is completely porous to achiral glycine. Hence, L-chiral rejection is the only design principle on which DTD functions, unlike other chiral-specific enzymes such as D-amino acid oxidases, which are specific for D-enantiomers. Competition assays with elongation factor thermo unstable (EF-Tu and DTD demonstrate that EF-Tu precludes Gly-tRNAGly misediting at normal cellular concentrations. However, even slightly higher DTD levels overcome this protection conferred by EF-Tu, thus resulting in significant depletion of Gly-tRNAGly. Our in vitro observations are substantiated by cell-based studies in Escherichia coli that show that overexpression of DTD causes cellular toxicity, which is largely rescued upon glycine supplementation. Furthermore, we provide direct evidence that DTD is an RNA-based catalyst, since it uses only the terminal 2'-OH of tRNA for catalysis without the involvement of protein side chains. The study therefore provides a unique paradigm of enzyme action for substrate selection/specificity by DTD, and thus explains the underlying cause of DTD's activity on Gly-tRNAGly. It also gives a molecular and functional basis for the necessity and the observed tight regulation of DTD levels, thereby preventing cellular toxicity due to misediting.

  10. Blocking signaling at the level of GLI regulates downstream gene expression and inhibits proliferation of canine osteosarcoma cells.

    Shahi, Mehdi Hayat; Holt, Roseline; Rebhun, Robert B

    2014-01-01

    The Hedgehog-GLI signaling pathway is active in a variety of human malignancies and is known to contribute to the growth and survival of human osteosarcoma cells. In this study, we examined the expression and regulation of GLI transcription factors in multiple canine osteosarcoma cell lines and analyzed the effects of inhibiting GLI with GANT61, a GLI-specific inhibitor. Compared with normal canine osteoblasts, real-time PCR showed that GLI1 and GLI2 were highly expressed in two out of three cell lines and correlated with downstream target gene expression of PTCH1and PAX6. Treatment of canine osteosarcoma cells with GANT61 resulted in decreased expression of GLI1, GLI2, PTCH1, and PAX6. Furthermore, GANT61 inhibited proliferation and colony formation in all three canine osteosarcoma cell lines. The finding that GLI signaling activity is present and active in canine osteosarcoma cells suggests that spontaneously arising osteosarcoma in dogs might serve as a good model for future preclinical testing of GLI inhibitors.

  11. Blocking signaling at the level of GLI regulates downstream gene expression and inhibits proliferation of canine osteosarcoma cells.

    Mehdi Hayat Shahi

    Full Text Available The Hedgehog-GLI signaling pathway is active in a variety of human malignancies and is known to contribute to the growth and survival of human osteosarcoma cells. In this study, we examined the expression and regulation of GLI transcription factors in multiple canine osteosarcoma cell lines and analyzed the effects of inhibiting GLI with GANT61, a GLI-specific inhibitor. Compared with normal canine osteoblasts, real-time PCR showed that GLI1 and GLI2 were highly expressed in two out of three cell lines and correlated with downstream target gene expression of PTCH1and PAX6. Treatment of canine osteosarcoma cells with GANT61 resulted in decreased expression of GLI1, GLI2, PTCH1, and PAX6. Furthermore, GANT61 inhibited proliferation and colony formation in all three canine osteosarcoma cell lines. The finding that GLI signaling activity is present and active in canine osteosarcoma cells suggests that spontaneously arising osteosarcoma in dogs might serve as a good model for future preclinical testing of GLI inhibitors.

  12. Abrogation of Gli3 expression suppresses the growth of colon cancer cells via activation of p53

    Kang, Han Na; Oh, Sang Cheul; Kim, Jun Suk; Yoo, Young A.

    2012-01-01

    p53, the major human tumor suppressor, appears to be related to sonic hedgehog (Shh)–Gli-mediated tumorigenesis. However, the role of p53 in tumor progression by the Shh–Gli signaling pathway is poorly understood. Herein we investigated the critical regulation of Gli3–p53 in tumorigenesis of colon cancer cells and the molecular mechanisms underlying these effects. RT-PCR analysis indicated that the mRNA level of Shh and Gli3 in colon tumor tissues was significantly higher than corresponding normal tissues (P < 0.001). The inhibition of Gli3 by treatment with Gli3 siRNA resulted in a clear decrease in cell proliferation and enhanced the level of expression of p53 proteins compared to treatment with control siRNA. The half-life of p53 was dramatically increased by treatment with Gli3 siRNA. In addition, treatment with MG132 blocked MDM2-mediated p53 ubiquitination and degradation, and led to accumulation of p53 in Gli3 siRNA-overexpressing cells. Importantly, ectopic expression of p53 siRNA reduced the ability of Gli3 siRNA to suppress proliferation of those cells compared with the cells treated with Gli3 siRNA alone. Moreover, Gli3 siRNA sensitized colon cancer cells to treatment with anti-cancer agents (5-FU and bevacizumab). Taken together, our studies demonstrate that loss of Gli3 signaling leads to disruption of the MDM2–p53 interaction and strongly potentiate p53-dependent cell growth inhibition in colon cancer cells, indicating a basis for the rational use of Gli3 antagonists as a novel treatment option for colon cancer.

  13. Codon 201Gly Polymorphic Type of the DCC Gene is Related to Disseminated Neuroblastoma

    Xiao-Tang Kong

    2001-01-01

    Full Text Available The deleted in colorectal carcinoma (DCC gene is a potential tumor- suppressor gene on chromosome 18821.3. The relatively high frequency of loss of heterozygosity (LOH and loss of expression of this gene in neuroblastoma, especially in the advanced stages, imply the possibility of involvement of the DCC gene in progression of neuroblastoma. However, only few typical mutations have been identified in this gene, indicating that other possible mechanisms for the inactivation of this gene may exist. A polymorphic change (Arg to Gly at DCC codon 201 is related to advanced colorectal carcinoma and increases in the tumors with absent DCC protein expression. In order to understand whether this change is associated with the development or progression of neuroblastoma, we investigated codon 201 polymorphism of the DCC gene in 102 primary neuroblastomas by polymerase chain reaction single-strand conformation polymorphism. We found no missense or nonsense mutations, but a polymorphic change from CGA (Arg to GGA (Gly at codon 201 resulting in three types of polymorphism: codon 201Gly type, codon 201Arg/Gly type, and codon 201Arg type. The codon 201Gly type occurred more frequently in disseminated (stages IV and IVs neuroblastomas (72% than in localized (stages I, II, and III tumors (48% (P=.035, and normal controls (38% (P=.024. In addition, the codon 201Gly type was significantly more common in tumors found clinically (65% than in those found by mass screening (35% (P=.002. The results suggested that the codon 201Gly type of the DCC gene might be associated with a higher risk of disseminating neuroblastoma.

  14. Steroidal regulation of Ihh and Gli1 expression in the rat uterus.

    Kubota, Kaiyu; Yamauchi, Nobuhiko; Yamagami, Kazuki; Nishimura, Sho; Gobaru, Takafumi; Yamanaka, Ken-ichi; Wood, Chris; Soh, Tomoki; Takahashi, Masashi; Hattori, Masa-aki

    2010-05-01

    Ovarian steroid hormones, progesterone (P4), and estradiol (E2) strictly regulate the endometrial tissue remodeling required for successful embryo implantation. Indian hedgehog (Ihh) is up-regulated by P4 and critically mediates uterine receptivity in the mouse. However, the regulation of Ihh expression during the implantation period still remains unclear. The present study was conducted to elucidate the mechanism of the steroidal regulation in the expression of Ihh and Gli1, the mediator of the Ihh pathway. Ihh mRNA was expressed in the rat uterus on 3.5-5.5 days post-coitus (dpc), while Gli1 expression transiently increased at 3.5 dpc but decreased significantly on 5.5 dpc (P Ihh was induced by the implantation-induced E2 treatment in the primed rat uterus. In contrast, expression of Gli1 was significantly decreased by E2 treatment (P = 0.016). In the case of ICI182.780 (ICI) treatment, Ihh expression was eliminated by ICI, whilst Gli1 expression increased. These results suggest that Ihh expression is maintained at a high level until the initiation of implantation, while the expression of Gli1 is decreased just prior to the initiation of implantation depending on the E2 action. This observation aids in the understanding of the Ihh signaling pathway mediating uterine remodeling for implantation.

  15. Structural basis of SUFU–GLI interaction in human Hedgehog signalling regulation

    Cherry, Amy L.; Finta, Csaba; Karlström, Mikael; Jin, Qianren; Schwend, Thomas; Astorga-Wells, Juan; Zubarev, Roman A.; Del Campo, Mark; Criswell, Angela R.; Sanctis, Daniele de; Jovine, Luca; Toftgård, Rune

    2013-01-01

    Crystal and small-angle X-ray scattering structures of full-length human SUFU alone and in complex with the conserved SYGHL motif from GLI transcription factors show major conformational changes associated with binding and reveal an intrinsically disordered region crucial for pathway activation. Hedgehog signalling plays a fundamental role in the control of metazoan development, cell proliferation and differentiation, as highlighted by the fact that its deregulation is associated with the development of many human tumours. SUFU is an essential intracellular negative regulator of mammalian Hedgehog signalling and acts by binding and modulating the activity of GLI transcription factors. Despite its central importance, little is known about SUFU regulation and the nature of SUFU–GLI interaction. Here, the crystal and small-angle X-ray scattering structures of full-length human SUFU and its complex with the key SYGHL motif conserved in all GLIs are reported. It is demonstrated that GLI binding is associated with major conformational changes in SUFU, including an intrinsically disordered loop that is also crucial for pathway activation. These findings reveal the structure of the SUFU–GLI interface and suggest a mechanism for an essential regulatory step in Hedgehog signalling, offering possibilities for the development of novel pathway modulators and therapeutics

  16. Structural basis of SUFU–GLI interaction in human Hedgehog signalling regulation

    Cherry, Amy L.; Finta, Csaba; Karlström, Mikael; Jin, Qianren; Schwend, Thomas [Karolinska Institutet, Novum, Hälsovägen 7, SE-141 83 Huddinge (Sweden); Astorga-Wells, Juan [Karolinska Institutet, Scheeles väg 2, SE-171 77 Stockholm (Sweden); Biomotif AB, Enhagsvägen 7, SE-182 12 Danderyd (Sweden); Zubarev, Roman A. [Karolinska Institutet, Scheeles väg 2, SE-171 77 Stockholm (Sweden); Del Campo, Mark; Criswell, Angela R. [Rigaku Americas Corporation, 9009 New Trails Drive, The Woodlands, TX 77381 (United States); Sanctis, Daniele de [European Synchrotron Radiation Facility, 6 Rue Jules Horowitz, 38043 Grenoble (France); Jovine, Luca, E-mail: luca.jovine@ki.se; Toftgård, Rune, E-mail: luca.jovine@ki.se [Karolinska Institutet, Novum, Hälsovägen 7, SE-141 83 Huddinge (Sweden)

    2013-12-01

    Crystal and small-angle X-ray scattering structures of full-length human SUFU alone and in complex with the conserved SYGHL motif from GLI transcription factors show major conformational changes associated with binding and reveal an intrinsically disordered region crucial for pathway activation. Hedgehog signalling plays a fundamental role in the control of metazoan development, cell proliferation and differentiation, as highlighted by the fact that its deregulation is associated with the development of many human tumours. SUFU is an essential intracellular negative regulator of mammalian Hedgehog signalling and acts by binding and modulating the activity of GLI transcription factors. Despite its central importance, little is known about SUFU regulation and the nature of SUFU–GLI interaction. Here, the crystal and small-angle X-ray scattering structures of full-length human SUFU and its complex with the key SYGHL motif conserved in all GLIs are reported. It is demonstrated that GLI binding is associated with major conformational changes in SUFU, including an intrinsically disordered loop that is also crucial for pathway activation. These findings reveal the structure of the SUFU–GLI interface and suggest a mechanism for an essential regulatory step in Hedgehog signalling, offering possibilities for the development of novel pathway modulators and therapeutics.

  17. Regulation of Calvarial Osteogenesis by Concomitant De-repression of GLI3 and Activation of IHH Targets

    Lotta K. Veistinen; Tuija Mustonen; Tuija Mustonen; Md. Rakibul Hasan; Maarit Takatalo; Yukiho Kobayashi; Yukiho Kobayashi; Dörthe A. Kesper; Andrea Vortkamp; David P. Rice; David P. Rice

    2017-01-01

    Loss-of-function mutations in GLI3 and IHH cause craniosynostosis and reduced osteogenesis, respectively. In this study, we show that Ihh ligand, the receptor Ptch1 and Gli transcription factors are differentially expressed in embryonic mouse calvaria osteogenic condensations. We show that in both Ihh−/− and Gli3Xt−J/Xt−J embryonic mice, the normal gene expression architecture is lost and this results in disorganized calvarial bone development. RUNX2 is a master regulatory transcription facto...

  18. A mechanism for vertebrate Hedgehog signaling: recruitment to cilia and dissociation of SuFu-Gli protein complexes.

    Tukachinsky, Hanna; Lopez, Lyle V; Salic, Adrian

    2010-10-18

    In vertebrates, Hedgehog (Hh) signaling initiated in primary cilia activates the membrane protein Smoothened (Smo) and leads to activation of Gli proteins, the transcriptional effectors of the pathway. In the absence of signaling, Gli proteins are inhibited by the cytoplasmic protein Suppressor of Fused (SuFu). It is unclear how Hh activates Gli and whether it directly regulates SuFu. We find that Hh stimulation quickly recruits endogenous SuFu-Gli complexes to cilia, suggesting a model in which Smo activates Gli by relieving inhibition by SuFu. In support of this model, we find that Hh causes rapid dissociation of the SuFu-Gli complex, thus allowing Gli to enter the nucleus and activate transcription. Activation of protein kinase A (PKA), an inhibitor of Hh signaling, blocks ciliary localization of SuFu-Gli complexes, which in turn prevents their dissociation by signaling. Our results support a simple mechanism in which Hh signals at vertebrate cilia cause dissociation of inactive SuFu-Gli complexes, a process inhibited by PKA.

  19. A mechanism for vertebrate Hedgehog signaling: recruitment to cilia and dissociation of SuFu–Gli protein complexes

    Tukachinsky, Hanna; Lopez, Lyle V.

    2010-01-01

    In vertebrates, Hedgehog (Hh) signaling initiated in primary cilia activates the membrane protein Smoothened (Smo) and leads to activation of Gli proteins, the transcriptional effectors of the pathway. In the absence of signaling, Gli proteins are inhibited by the cytoplasmic protein Suppressor of Fused (SuFu). It is unclear how Hh activates Gli and whether it directly regulates SuFu. We find that Hh stimulation quickly recruits endogenous SuFu–Gli complexes to cilia, suggesting a model in which Smo activates Gli by relieving inhibition by SuFu. In support of this model, we find that Hh causes rapid dissociation of the SuFu–Gli complex, thus allowing Gli to enter the nucleus and activate transcription. Activation of protein kinase A (PKA), an inhibitor of Hh signaling, blocks ciliary localization of SuFu–Gli complexes, which in turn prevents their dissociation by signaling. Our results support a simple mechanism in which Hh signals at vertebrate cilia cause dissociation of inactive SuFu–Gli complexes, a process inhibited by PKA. PMID:20956384

  20. Lithium inhibits tumorigenic potential of PDA cells through targeting hedgehog-GLI signaling pathway.

    Zhonglu Peng

    Full Text Available Hedgehog signaling pathway plays a critical role in the initiation and development of pancreatic ductal adenocarcinoma (PDA and represents an attractive target for PDA treatment. Lithium, a clinical mood stabilizer for mental disorders, potently inhibits the activity of glycogen synthase kinase 3β (GSK3β that promotes the ubiquitin-dependent proteasome degradation of GLI1, an important downstream component of hedgehog signaling. Herein, we report that lithium inhibits cell proliferation, blocks G1/S cell-cycle progression, induces cell apoptosis and suppresses tumorigenic potential of PDA cells through down-regulation of the expression and activity of GLI1. Moreover, lithium synergistically enhances the anti-cancer effect of gemcitabine. These findings further our knowledge of mechanisms of action for lithium and provide a potentially new therapeutic strategy for PDA through targeting GLI1.

  1. Mutation of the conserved Gly94 and Gly126 in elongation factor Tu from Escherichia coli. Elucidation of their structural and functional roles

    Knudsen, Charlotte Rohde; Kjaersgård, I V; Wiborg, O

    1995-01-01

    All guanine-nucleotide-binding proteins cycle between an inactive, GDP-bound and an active, GTP-bound conformation whereby they function as molecular switches. Elongation factor Tu from Escherichia coli is used as a model for defining residues important in the switch mechanism. Gly94 and Gly126...... were separately mutated to alanine residues to study their role in the switch mechanism. The mutant proteins are denoted [G94A]EF-Tu and [G126A]EF-Tu, respectively. Both mutations affect the affinities for guanine nucleotides considerably, resulting in a decrease in the characteristic preference...... for GDP over GTP. Furthermore the [G94A]EF-Tu mutant possesses an increased GTPase activity. The aminoacyl-tRNA affinity is much reduced for [G94A]EF-Tu, as reflected in an increase of the dissociation rate constant for the ternary complex by a factor of 40. Surprisingly, however, both mutants...

  2. Covalent attachment of cell-adhesive peptide Gly-Arg-Gly-Asp (GRGD) to poly(etheretherketone) surface by tailored silanization layers technique

    Zheng, Yanyan; Xiong, Chengdong; Li, Xiaoyu; Zhang, Lifang

    2014-01-01

    Highlights: • The carbonyl groups on PEEK surface were effectively reduced to hydroxyl groups using sodium borohydride. • Silanization layers technique was employed to immobilize the cell-adhesive peptide Gly-Arg-Gly-Asp (GRGD) on hydroxylation-pretreated PEEK sheet surface by covalent chemical attachment. • XPS, surface profiler and water contact angle measurements proved the presence of GRGD on PEEK surface. • Osteoblast-like cells (MC3T3-E1) attachment and proliferation were improved effectively on GRGD-modified PEEK surface. - Abstract: Poly(etheretherketone) (PEEK) is a rigid semicrystalline polymer that combines excellent mechanical properties, broad chemical resistance and bone-like stiffness and is widely used in biomedical fields. However, PEEK is naturally bioinert, leading to limited biomedical applications, especially when a direct bone-implant osteointegration is desired. In this study, a three-step reaction procedure was employed to immobilize the cell-adhesive peptide Gly-Arg-Gly-Asp (GRGD) on the surface of PEEK sheet by covalent chemical attachment to favor cell adhesion and proliferation. First, hydroxylation-pretreated PEEK surfaces were silanized with 7-Oct-1-enyltrichlorosilane (OETS) in dry cyclohexane, resulting in a silanization layer with terminal ethenyl. Second, the terminal ethylenic double bonds of the silanization layer on PEEK surface were converted to carboxyl groups through acidic potassium manganate oxidation. Finally, GRGD was covalently attached by carbodiimide mediated condensation between the carboxyl on PEEK surface and amine presents in GRGD. X-ray photoelectron spectroscopy (XPS), attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy, surface profiler and water contact angle measurements were applied to characterize the modified surfaces. The effect of cells attachment and proliferation on each specimen was investigated. Pre-osteoblast cells (MC3T3-E1) attachment, spreading and proliferation

  3. GlyGly-CTERM and rhombosortase: a C-terminal protein processing signal in a many-to-one pairing with a rhomboid family intramembrane serine protease.

    Daniel H Haft

    Full Text Available The rhomboid family of serine proteases occurs in all domains of life. Its members contain at least six hydrophobic membrane-spanning helices, with an active site serine located deep within the hydrophobic interior of the plasma membrane. The model member GlpG from Escherichia coli is heavily studied through engineered mutant forms, varied model substrates, and multiple X-ray crystal studies, yet its relationship to endogenous substrates is not well understood. Here we describe an apparent membrane anchoring C-terminal homology domain that appears in numerous genera including Shewanella, Vibrio, Acinetobacter, and Ralstonia, but excluding Escherichia and Haemophilus. Individual genomes encode up to thirteen members, usually homologous to each other only in this C-terminal region. The domain's tripartite architecture consists of motif, transmembrane helix, and cluster of basic residues at the protein C-terminus, as also seen with the LPXTG recognition sequence for sortase A and the PEP-CTERM recognition sequence for exosortase. Partial Phylogenetic Profiling identifies a distinctive rhomboid-like protease subfamily almost perfectly co-distributed with this recognition sequence. This protease subfamily and its putative target domain are hereby renamed rhombosortase and GlyGly-CTERM, respectively. The protease and target are encoded by consecutive genes in most genomes with just a single target, but far apart otherwise. The signature motif of the Rhombo-CTERM domain, often SGGS, only partially resembles known cleavage sites of rhomboid protease family model substrates. Some protein families that have several members with C-terminal GlyGly-CTERM domains also have additional members with LPXTG or PEP-CTERM domains instead, suggesting there may be common themes to the post-translational processing of these proteins by three different membrane protein superfamilies.

  4. Covalent attachment of cell-adhesive peptide Gly-Arg-Gly-Asp (GRGD) to poly(etheretherketone) surface by tailored silanization layers technique

    Zheng, Yanyan [Chengdu Institute of Organic Chemistry, Chinese Academy of Sciences, Chengdu 610041 (China); University of Chinese Academy of Sciences, Beijing 100049 (China); Xiong, Chengdong [Chengdu Institute of Organic Chemistry, Chinese Academy of Sciences, Chengdu 610041 (China); Li, Xiaoyu [State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Zhang, Lifang, E-mail: zhanglfcioc@163.com [Chengdu Institute of Organic Chemistry, Chinese Academy of Sciences, Chengdu 610041 (China)

    2014-11-30

    Highlights: • The carbonyl groups on PEEK surface were effectively reduced to hydroxyl groups using sodium borohydride. • Silanization layers technique was employed to immobilize the cell-adhesive peptide Gly-Arg-Gly-Asp (GRGD) on hydroxylation-pretreated PEEK sheet surface by covalent chemical attachment. • XPS, surface profiler and water contact angle measurements proved the presence of GRGD on PEEK surface. • Osteoblast-like cells (MC3T3-E1) attachment and proliferation were improved effectively on GRGD-modified PEEK surface. - Abstract: Poly(etheretherketone) (PEEK) is a rigid semicrystalline polymer that combines excellent mechanical properties, broad chemical resistance and bone-like stiffness and is widely used in biomedical fields. However, PEEK is naturally bioinert, leading to limited biomedical applications, especially when a direct bone-implant osteointegration is desired. In this study, a three-step reaction procedure was employed to immobilize the cell-adhesive peptide Gly-Arg-Gly-Asp (GRGD) on the surface of PEEK sheet by covalent chemical attachment to favor cell adhesion and proliferation. First, hydroxylation-pretreated PEEK surfaces were silanized with 7-Oct-1-enyltrichlorosilane (OETS) in dry cyclohexane, resulting in a silanization layer with terminal ethenyl. Second, the terminal ethylenic double bonds of the silanization layer on PEEK surface were converted to carboxyl groups through acidic potassium manganate oxidation. Finally, GRGD was covalently attached by carbodiimide mediated condensation between the carboxyl on PEEK surface and amine presents in GRGD. X-ray photoelectron spectroscopy (XPS), attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy, surface profiler and water contact angle measurements were applied to characterize the modified surfaces. The effect of cells attachment and proliferation on each specimen was investigated. Pre-osteoblast cells (MC3T3-E1) attachment, spreading and proliferation

  5. GLI1 is a central mediator of EWS/FLI1 signaling in Ewing tumors.

    Jay Joo

    2009-10-01

    Full Text Available The Ewing Sarcoma Family Tumors (ESFT consist of the classical pathologic entities of Ewing Sarcoma and peripheral Primitive Neuroectodermal Tumor. Occurring largely in the childhood through young adult years, these tumors have an unsurpassed propensity for metastasis and have no defined cell of origin. The biology of these aggressive malignancies centers around EWS/FLI1 and related EWS/ETS chimeric transcription factors, which are largely limited to this tumor class. Much progress has been made in the identification of a network of loci whose expression is modulated by EWS/FLI1 and its congeners. To date, little progress has been made in reconstructing the sequence of direct and indirect events that produce this network of modulated loci. The recent identification of GLI1 as an upregulated target of EWS/ETS transcription factors suggests a target which may be a more central mediator in the ESFT signaling network. In this paper, we further define the relationship of EWS/FLI1 expression and GLI1 upregulation in ESFT. This relationship is supported with data from primary tumor specimens. It is consistently observed across multiple ESFT cell lines and with multiple means of EWS/FLI1 inhibition. GLI1 inhibition affects tumor cell line phenotype whether shRNA or endogenous or pharmacologic inhibitors are employed. As is seen in model transformation systems, GLI1 upregulation by EWS/FLI1 appears to be independent of Hedgehog stimulation. Consistent with a more central role in ESFT pathogenesis, several known EWS/FLI1 targets appear to be targeted through GLI1. These findings further establish a central role for GLI1 in the pathogenesis of Ewing Tumors.

  6. Maintenance of Taste Organs Is Strictly Dependent on Epithelial Hedgehog/GLI Signaling.

    Alexandre N Ermilov

    2016-11-01

    Full Text Available For homeostasis, lingual taste papilla organs require regulation of epithelial cell survival and renewal, with sustained innervation and stromal interactions. To investigate a role for Hedgehog/GLI signaling in adult taste organs we used a panel of conditional mouse models to manipulate GLI activity within epithelial cells of the fungiform and circumvallate papillae. Hedgehog signaling suppression rapidly led to taste bud loss, papilla disruption, and decreased proliferation in domains of papilla epithelium that contribute to taste cells. Hedgehog responding cells were eliminated from the epithelium but retained in the papilla stromal core. Despite papilla disruption and loss of taste buds that are a major source of Hedgehog ligand, innervation to taste papillae was maintained, and not misdirected, even after prolonged GLI blockade. Further, vimentin-positive fibroblasts remained in the papilla core. However, retained innervation and stromal cells were not sufficient to maintain taste bud cells in the context of compromised epithelial Hedgehog signaling. Importantly taste organ disruption after GLI blockade was reversible in papillae that retained some taste bud cell remnants where reactivation of Hedgehog signaling led to regeneration of papilla epithelium and taste buds. Therefore, taste bud progenitors were either retained during epithelial GLI blockade or readily repopulated during recovery, and were poised to regenerate taste buds once Hedgehog signaling was restored, with innervation and papilla connective tissue elements in place. Our data argue that Hedgehog signaling is essential for adult tongue tissue maintenance and that taste papilla epithelial cells represent the key targets for physiologic Hedgehog-dependent regulation of taste organ homeostasis. Because disruption of GLI transcriptional activity in taste papilla epithelium is sufficient to drive taste organ loss, similar to pharmacologic Hedgehog pathway inhibition, the findings

  7. Maintenance of Taste Organs Is Strictly Dependent on Epithelial Hedgehog/GLI Signaling.

    Ermilov, Alexandre N; Kumari, Archana; Li, Libo; Joiner, Ariell M; Grachtchouk, Marina A; Allen, Benjamin L; Dlugosz, Andrzej A; Mistretta, Charlotte M

    2016-11-01

    For homeostasis, lingual taste papilla organs require regulation of epithelial cell survival and renewal, with sustained innervation and stromal interactions. To investigate a role for Hedgehog/GLI signaling in adult taste organs we used a panel of conditional mouse models to manipulate GLI activity within epithelial cells of the fungiform and circumvallate papillae. Hedgehog signaling suppression rapidly led to taste bud loss, papilla disruption, and decreased proliferation in domains of papilla epithelium that contribute to taste cells. Hedgehog responding cells were eliminated from the epithelium but retained in the papilla stromal core. Despite papilla disruption and loss of taste buds that are a major source of Hedgehog ligand, innervation to taste papillae was maintained, and not misdirected, even after prolonged GLI blockade. Further, vimentin-positive fibroblasts remained in the papilla core. However, retained innervation and stromal cells were not sufficient to maintain taste bud cells in the context of compromised epithelial Hedgehog signaling. Importantly taste organ disruption after GLI blockade was reversible in papillae that retained some taste bud cell remnants where reactivation of Hedgehog signaling led to regeneration of papilla epithelium and taste buds. Therefore, taste bud progenitors were either retained during epithelial GLI blockade or readily repopulated during recovery, and were poised to regenerate taste buds once Hedgehog signaling was restored, with innervation and papilla connective tissue elements in place. Our data argue that Hedgehog signaling is essential for adult tongue tissue maintenance and that taste papilla epithelial cells represent the key targets for physiologic Hedgehog-dependent regulation of taste organ homeostasis. Because disruption of GLI transcriptional activity in taste papilla epithelium is sufficient to drive taste organ loss, similar to pharmacologic Hedgehog pathway inhibition, the findings suggest that taste

  8. Hedgehog-GLI signaling drives self-renewal and tumorigenicity of human melanoma-initiating cells.

    Santini, Roberta; Vinci, Maria C; Pandolfi, Silvia; Penachioni, Junia Y; Montagnani, Valentina; Olivito, Biagio; Gattai, Riccardo; Pimpinelli, Nicola; Gerlini, Gianni; Borgognoni, Lorenzo; Stecca, Barbara

    2012-09-01

    The question of whether cancer stem/tumor-initiating cells (CSC/TIC) exist in human melanomas has arisen in the last few years. Here, we have used nonadherent spheres and the aldehyde dehydrogenase (ALDH) enzymatic activity to enrich for CSC/TIC in a collection of human melanomas obtained from a broad spectrum of sites and stages. We find that melanomaspheres display extensive in vitro self-renewal ability and sustain tumor growth in vivo, generating human melanoma xenografts that recapitulate the phenotypic composition of the parental tumor. Melanomaspheres express high levels of Hedgehog (HH) pathway components and of embryonic pluripotent stem cell factors SOX2, NANOG, OCT4, and KLF4. We show that human melanomas contain a subset of cells expressing high ALDH activity (ALDH(high)), which is endowed with higher self-renewal and tumorigenic abilities than the ALDH(low) population. A good correlation between the number of ALDH(high) cells and sphere formation efficiency was observed. Notably, both pharmacological inhibition of HH signaling by the SMOOTHENED (SMO) antagonist cyclopamine and GLI antagonist GANT61 and stable expression of shRNA targeting either SMO or GLI1 result in a significant decrease in melanoma stem cell self-renewal in vitro and a reduction in the number of ALDH(high) melanoma stem cells. Finally, we show that interference with the HH-GLI pathway through lentiviral-mediated silencing of SMO and GLI1 drastically diminishes tumor initiation of ALDH(high) melanoma stem cells. In conclusion, our data indicate an essential role of the HH-GLI1 signaling in controlling self-renewal and tumor initiation of melanoma CSC/TIC. Targeting HH-GLI1 is thus predicted to reduce the melanoma stem cell compartment. Copyright © 2012 AlphaMed Press.

  9. The transcription factor GLI1 modulates the inflammatory response during pancreatic tissue remodeling.

    Mathew, Esha; Collins, Meredith A; Fernandez-Barrena, Maite G; Holtz, Alexander M; Yan, Wei; Hogan, James O; Tata, Zachary; Allen, Benjamin L; Fernandez-Zapico, Martin E; di Magliano, Marina Pasca

    2014-10-03

    Pancreatic cancer, one of the deadliest human malignancies, is almost uniformly associated with a mutant, constitutively active form of the oncogene Kras. Studies in genetically engineered mouse models have defined a requirement for oncogenic KRAS in both the formation of pancreatic intraepithelial neoplasias, the most common precursor lesions to pancreatic cancer, and in the maintenance and progression of these lesions. Previous work using an inducible model allowing tissue-specific and reversible expression of oncogenic Kras in the pancreas indicates that inactivation of this GTPase at the pancreatic intraepithelial neoplasia stage promotes pancreatic tissue repair. Here, we extend these findings to identify GLI1, a transcriptional effector of the Hedgehog pathway, as a central player in pancreatic tissue repair upon Kras inactivation. Deletion of a single allele of Gli1 results in improper stromal remodeling and perdurance of the inflammatory infiltrate characteristic of pancreatic tumorigenesis. Strikingly, this partial loss of Gli1 affects activated fibroblasts in the pancreas and the recruitment of immune cells that are vital for tissue recovery. Analysis of the mechanism using expression and chromatin immunoprecipitation assays identified a subset of cytokines, including IL-6, mIL-8, Mcp-1, and M-csf (Csf1), as direct GLI1 target genes potentially mediating this phenomenon. Finally, we demonstrate that canonical Hedgehog signaling, a known regulator of Gli1 activity, is required for pancreas recovery. Collectively, these data delineate a new pathway controlling tissue repair and highlight the importance of GLI1 in regulation of the pancreatic microenvironment during this cellular process. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  10. Gli Ungulati in Italia: status, gestione e ricerca scientifica

    Marco Apollonio

    2004-06-01

    managers in public administrations is suggested, as a further step in the development of a proper management of ungulates and other wildlife. Scientific research on ungulates in Italy was, in the last decade, quite uneven both in the species and in the disciplines chosen. There is a clear opportunistic approach to subjects, that often has no link with the real necessities of wildlife management. The development of long term studies is suggested, together with a more efficient coordination among scientist in the choice of the research subjects. Last but not least a more efficient founding mechanisms is needed in order to avoid money waste and to promote serious research programmes. Riassunto Gli Ungulati nel nostro Paese hanno proseguito, nell'ultimo decennio, l'espansione dei loro areali già iniziata nel decennio precedente e, per quanto i dati disponibili possono testimoniare, della loro consistenza. Tale tendenza è particolarmente chiara nel Centro-Nord, mentre nell'area meridionale il processo di sviluppo di popolamenti di ungulati delle diverse specie è rallentato se non assente, sia pure con notevoli eccezioni come la Sardegna dove, grazie ad un'opportuna politica di reintroduzioni, il Cervo sardo (Cervus elaphus corsicanus sta tornando nelle quattro province dell'isola. In questo quadro generale deve essere ricordato il ruolo rilevante dell'aumento degli ungulati nel favorire l'espansione dei grandi predatori, in particolare del Lupo (Canis lupus in ampi settori del paese. La gestione degli ungulati ha anch'essa subito una positiva evoluzione durante l'ultimo decennio sebbene persistano forti differenze fra Centro-Nord e Sud. Reintroduzioni di molte specie, effettuate con procedure nel complesso più corrette di quelle eseguite nel passato, hanno contribuito a colmare le lacune della distribuzione naturale di molte specie. Le statistiche d'abbattimento disponibili hanno dimensioni che suggeriscono un'accresciuta importanza degli

  11. Gli3 acts as a repressor downstream of Ihh in regulating two distinct steps of chondrocyte differentiation.

    Koziel, Lydia; Wuelling, Manuela; Schneider, Sabine; Vortkamp, Andrea

    2005-12-01

    During endochondral ossification, the secreted growth factor Indian hedgehog (Ihh) regulates several differentiation steps. It interacts with a second secreted factor, parathyroid hormone-related protein (PTHrP), to regulate the onset of hypertrophic differentiation, and it regulates chondrocyte proliferation and ossification of the perichondrium independently of PTHrP. To investigate how the Ihh signal is translated in the different target tissues, we analyzed the role of the zinc-finger transcription factor Gli3, which acts downstream of hedgehog signals in other organs. Loss of Gli3 in Ihh mutants restores chondrocyte proliferation and delays the accelerated onset of hypertrophic differentiation observed in Ihh-/- mutants. Furthermore the expression of the Ihh target genes patched (Ptch) and PTHrP is reactivated in Ihh-/-;Gli3-/- mutants. Gli3 seems thus to act as a strong repressor of Ihh signals in regulating chondrocyte differentiation. In addition, loss of Gli3 in mice that overexpress Ihh in chondrocytes accelerates the onset of hypertrophic differentiation by reducing the domain and possibly the level of PTHrP expression. Careful analysis of chondrocyte differentiation in Gli3-/- mutants revealed that Gli3 negatively regulates the differentiation of distal, low proliferating chondrocytes into columnar, high proliferating cells. Our results suggest a model in which the Ihh/Gli3 system regulates two distinct steps of chondrocyte differentiation: (1) the switch from distal into columnar chondrocytes is repressed by Gli3 in a PTHrP-independent mechanism; (2) the transition from proliferating into hypertrophic chondrocytes is regulated by Gli3-dependent expression of PTHrP. Furthermore, by regulating distal chondrocyte differentiation, Gli3 seems to position the domain of PTHrP expression.

  12. Identification of GLI Mutations in Patients With Hirschsprung Disease That Disrupt Enteric Nervous System Development in Mice.

    Liu, Jessica Ai-Jia; Lai, Frank Pui-Ling; Gui, Hong-Sheng; Sham, Mai-Har; Tam, Paul Kwong-Hang; Garcia-Barcelo, Maria-Mercedes; Hui, Chi-Chung; Ngan, Elly Sau-Wai

    2015-12-01

    Hirschsprung disease is characterized by a deficit in enteric neurons, which are derived from neural crest cells (NCCs). Aberrant hedgehog signaling disrupts NCC differentiation and might cause Hirschsprung disease. We performed genetic analyses to determine whether hedgehog signaling is involved in pathogenesis. We performed deep-target sequencing of DNA from 20 patients with Hirschsprung disease (16 men, 4 women), and 20 individuals without (controls), and searched for mutation(s) in GLI1, GLI2, GLI3, SUFU, and SOX10. Biological effects of GLI mutations were tested in luciferase reporter assays using HeLa or neuroblastoma cell lines. Development of the enteric nervous system was studied in Sufu(f/f), Gli3(Δ699), Wnt1-Cre, and Sox10(NGFP) mice using immunohistochemical and whole-mount staining procedures to quantify enteric neurons and glia and analyze axon fasciculation, respectively. NCC migration was studied using time-lapse imaging. We identified 3 mutations in GLI in 5 patients with Hirschsprung disease but no controls; all lead to increased transcription of SOX10 in cell lines. SUFU, GLI, and SOX10 form a regulatory loop that controls the neuronal vs glial lineages and migration of NCCs. Sufu mutants mice had high Gli activity, due to loss of Sufu, disrupting the regulatory loop and migration of enteric NCCs, leading to defective axonal fasciculation, delayed gut colonization, or intestinal hypoganglionosis. The ratio of enteric neurons to glia correlated inversely with Gli activity. We identified mutations that increase GLI activity in patients with Hirschsprung disease. Disruption of the SUFU-GLI-SOX10 regulatory loop disrupts migration of NCCs and development of the enteric nervous system in mice. Copyright © 2015 AGA Institute. Published by Elsevier Inc. All rights reserved.

  13. PKC-Dependent GlyT1 Ubiquitination Occurs Independent of Phosphorylation: Inespecificity in Lysine Selection for Ubiquitination.

    Susana P Barrera

    Full Text Available Neurotransmitter transporter ubiquitination is emerging as the main mechanism for endocytosis and sorting of cargo into lysosomes. In this study, we demonstrate PKC-dependent ubiquitination of three different isoforms of the glycine transporter 1 (GlyT1. Incubation of cells expressing transporter with the PKC activator phorbol ester induced a dramatic, time-dependent increase in GlyT1 ubiquitination, followed by accumulation of GlyT1 in EEA1 positive early endosomes. This occurred via a mechanism that was abolished by inhibition of PKC. GlyT1 endocytosis was confirmed in both retinal sections and primary cultures of mouse amacrine neurons. Replacement of only all lysines in the N-and C-termini to arginines prevented ubiquitination and endocytosis, displaying redundancy in the mechanism of ubiquitination. Interestingly, a 40-50% reduction in glycine uptake was detected in phorbol-ester stimulated cells expressing the WT-GlyT1, whereas no significant change was for the mutant protein, demonstrating that endocytosis participates in the reduction of uptake. Consistent with previous findings for the dopamine transporter DAT, ubiquitination of GlyT1 tails functions as sorting signal to deliver transporter into the lysosome and removal of ubiquitination sites dramatically attenuated the rate of GlyT1 degradation. Finally, we showed for the first time that PKC-dependent GlyT1 phosphorylation was not affected by removal of ubiquitination sites, suggesting separate PKC-dependent signaling events for these posttranslational modifications.

  14. Implications of the Transition From Zapletal to GLI Reference Values for Spirometry

    Raaijmakers, Lena; Zwitserloot, Annelies; Merkus, Peter; Gappa, Monika

    The current standard for monitoring lung function in children with asthma is spirometry. In Europe, results of these lung function tests have been related to Zapletal reference values published in 1977. Recently, the Global Lung Function Initiative (GLI) published predicted values of spirometry for

  15. Alpha-adducin Gly460Trp polymorphism and renal hemodynamics in essential hypertension

    Beeks, Esther; van der Klauw, Melanie M; Kroon, Abraham A; Spiering, Wilko; Fuss-Lejeune, Monique J M J; de Leeuw, Peter W

    2004-01-01

    Previous studies have shown an association between the alpha-adducin Gly460Trp polymorphism and salt-sensitive hypertension. Not much is known about the effects of the variants of this polymorphism on renal hemodynamics and function. Therefore, we performed the present study to investigate the

  16. Come diventare una Rockstar. Gli Houser e l’evoluzione del franchise Grand Theft Auto.

    Federico Giordano

    2013-03-01

    Full Text Available Wanted. La storia criminale di Grand Theft Auto, Multiplayer, 2012, il libro scritto da David Kushner, giornalista e docente universitario già autore del precedente, molto apprezzato, Master of Doom, è un testo importante per gli studi sui videogiochi, nonostante non si presenti in apparenza – a partire dal titolo evocativo – come testo accademico.

  17. Functional effects of the DCM mutant Gly159Asp troponin C in skinned muscle fibres

    Preston, Laura C; Lipscomb, Simon; Robinson, Paul

    2006-01-01

    We recently reported a dilated cardiomyopathy (DCM) causing mutation in a novel disease gene, TNNC1, which encodes cardiac troponin C (TnC). We have determined how this mutation, Gly159Asp, affects contractile regulation when incorporated into muscle fibres. Endogenous troponin in rabbit skinned...

  18. Stabilization of a chitinase from Serratia marcescens by Gly-->Ala and Xxx-->Pro mutations.

    Gaseidnes, S.; Synstad, B.; Jia, X.; Kjellesvik, H.; Vriend, G.; Eijsink, V.G.

    2003-01-01

    This paper describes attempts to increase the kinetic stability of chitinase B from Serratia marcescens (ChiB) by the introduction of semi-automatically designed rigidifying mutations of the Gly-->Ala and Xxx-->Pro type. Of 15 single mutants, several displayed significant increases in thermal

  19. GliZ, a transcriptional regulator of gliotoxin in Aspergillus fumigatus

    Bok, J.W.; Chung, D.W.; Balajee, A.

    2006-01-01

    Gliotoxin is a nonribosomal peptide produced by Aspergillus fumigatus. This compound has been proposed as an A. fumigatus virulence factor due to its cytotoxic, genotoxic, and apoptotic properties. Recent identification of the gliotoxin gene cluster identified several genes (gli genes) likely inv...

  20. GLI1, a master regulator of the hallmark of pancreatic cancer.

    Kasai, Kenji

    2016-12-01

    Hedgehog signaling is highly conserved across species and governs proper embryonic development. Germline gene mutations that reduce this signaling activity cause a variety of developmental abnormalities such as holoprosencephaly, while those that enhance Hedgehog signaling activity induce a tumor-predisposition condition Nevoid basal cell carcinoma syndrome. Furthermore, dysregulated activation of Hedgehog signaling has been recognized in various sporadic malignancies, including pancreatic adenocarcinoma. Pancreatic adenocarcinoma develops through a multistep carcinogenesis starting with oncogenic mutation of the KRAS gene. During this process, precancerous or cancer cells secrete Hedgehog ligand proteins to promote characteristic desmoplastic stroma around the cells, which in turn activates the expression of the downstream transcription factor GLI1 inside the cells. The quantitative and spatiotemporal dysregulation of GLI1 subsequently leads to the expression of transcriptional target genes of GLI1 that govern the hallmark of malignant properties. Here, after a brief introductory outline, a perspective is offered of Hedgehog signaling with a special focus on the role of GLI1 in pancreatic carcinogenesis. © 2016 Japanese Society of Pathology and John Wiley & Sons Australia, Ltd.

  1. Ihh controls cartilage development by antagonizing Gli3, but requires additional effectors to regulate osteoblast and vascular development.

    Hilton, Matthew J; Tu, Xiaolin; Cook, Julie; Hu, Hongliang; Long, Fanxin

    2005-10-01

    Indian hedgehog (Ihh) controls multiple aspects of endochondral skeletal development, including proliferation and maturation of chondrocytes, osteoblast development and cartilage vascularization. Although it is known that Gli transcription factors are key effectors of hedgehog signaling, it has not been established which Gli protein mediates Ihh activity in skeletal development. Here, we show that removal of Gli3 in Ihh-null mouse embryos restored normal proliferation and maturation of chondrocytes, but only partially rescued the defects in osteoblast development and cartilage vascularization. Remarkably, in both Ihh-/- and Ihh-/-; Gli3-/- embryos, vascularization promoted osteoblast development in perichondrial progenitor cells. Our results not only establish Gli3 as a critical effector for Ihh activity in the developing skeleton, but also identify an osteogenic role for a vasculature-derived signal, which integrates with Ihh and Wnt signals to determine the osteoblast versus chondrocyte fate in the mesenchymal progenitors.

  2. GLI1 is involved in cell cycle regulation and proliferation of NT2 embryonal carcinoma stem cells

    Vestergaard, Janni; Lind-Thomsen, Allan; Pedersen, Mikkel W.

    2008-01-01

    of altered HH signaling are interpreted by specific cell types. We have investigated the role of the HH transcription factor glioma-associated oncogene homolog 1 (GLI1) in the human Ntera2=D1 (NT2) embryonal carcinoma stem cell line. The study revealed that expression of GLI1 and its direct transcriptional......1 phase cyclins. In conclusion, our results suggest that GLI1 is involved in cell cycle and proliferation control in the embryonal carcinoma stem cell line NT2....... target Patched (PTCH) is downregulated in the early stages of retinoic acid-induced neuronal differentiation of NT2 cells. To identify transcriptional targets of the HH transcription factor GLI1 in NT2 cells, we performed global expression profiling following GLI1 RNA interference (RNAi). Of the similar...

  3. Expression of the glioma-associated oncogene homolog (GLI) 1 in human breast cancer is associated with unfavourable overall survival

    Haaf, Anette ten; Bektas, Nuran; Serenyi, Sonja von; Losen, Inge; Arweiler, Elfriede Christel; Hartmann, Arndt; Knüchel, Ruth; Dahl, Edgar

    2009-01-01

    The transcription factor GLI1, a member of the GLI subfamily of Krüppel-like zinc finger proteins is involved in signal transduction within the hedgehog pathway. Aberrant hedgehog signalling has been implicated in the development of different human tumour entities such as colon and lung cancer and increased GLI1 expression has been found in these tumour entities as well. In this study we questioned whether GLI1 expression might also be important in human breast cancer development. Furthermore we correlated GLI1 expression with histopathological and clinical data to evaluate whether GLI1 could represent a new prognostic marker in breast cancer treatment. Applying semiquantitative realtime PCR analysis and immunohistochemistry (IHC) GLI1 expression was analysed in human invasive breast carcinomas (n = 229) in comparison to normal human breast tissues (n = 58). GLI1 mRNA expression was furthermore analysed in a set of normal (n = 3) and tumourous breast cell lines (n = 8). IHC data were statistically interpreted using SPSS version 14.0. Initial analysis of GLI1 mRNA expression in a small cohort of (n = 5) human matched normal and tumourous breast tissues showed first tendency towards GLI1 overexpression in human breast cancers. However only a small sample number was included into these analyses and values for GLI1 overexpression were statistically not significant (P = 0.251, two-tailed Mann-Whitney U-test). On protein level, nuclear GLI1 expression in breast cancer cells was clearly more abundant than in normal breast epithelial cells (P = 0.008, two-tailed Mann-Whitney U-test) and increased expression of GLI1 protein in breast tumours significantly correlated with unfavourable overall survival (P = 0.019), but also with higher tumour stage (P < 0.001) and an increased number of tumour-positive axillar lymph nodes (P = 0.027). Interestingly, a highly significant correlation was found between GLI1 expression and the expression of SHH, a central upstream molecule of

  4. The topical penta-peptide Gly-Pro-Ile-Gly-Ser increases the proportion of thick hair in Japanese men with androgenetic alopecia.

    Iwabuchi, Tokuro; Takeda, Shunsuke; Yamanishi, Haruyo; Ideta, Ritsuro; Ehama, Ritsuko; Tsuruda, Akinori; Shibata, Hideaki; Ito, Tomoko; Komatsu, Nobuyuki; Terai, Keiko; Oka, Syuichi

    2016-06-01

    A penta-peptide, Gly-Pro-Ile-Gly-Ser (GPIGS), promotes proliferation of mouse hair keratinocytes and accelerates hair growth in mice. This study focused on the ability of the peptide to promote human hair growth. We used a human hair keratinocyte proliferation assay and organ cultures of human hair follicle as in vitro systems. The lotions with and without the penta-peptide were administered to 22 Japanese men with androgenetic alopecia (AGA) for 4 months in a double-blind and randomized clinical study. The penta-peptide significantly stimulated the proliferation of human hair keratinocytes at a concentration of 2.3 μm (P baldness (P = 0.020) when blinded reviewers graded photographs of the participants according to a standardized baldness scale. No adverse dermatological effects due to treatment were noted during this clinical study. This penta-peptide promotes proliferation of human hair keratinocytes and hair shaft elongation of human hair follicles, in vitro. This peptide increases thick hair ratio in vivo, and this compound is useful for the improvement of AGA. © 2016 Wiley Periodicals, Inc.

  5. Identification and Validation of Novel Hedgehog-Responsive Enhancers Predicted by Computational Analysis of Ci/Gli Binding Site Density

    Richards, Neil; Parker, David S.; Johnson, Lisa A.; Allen, Benjamin L.; Barolo, Scott; Gumucio, Deborah L.

    2015-01-01

    The Hedgehog (Hh) signaling pathway directs a multitude of cellular responses during embryogenesis and adult tissue homeostasis. Stimulation of the pathway results in activation of Hh target genes by the transcription factor Ci/Gli, which binds to specific motifs in genomic enhancers. In Drosophila, only a few enhancers (patched, decapentaplegic, wingless, stripe, knot, hairy, orthodenticle) have been shown by in vivo functional assays to depend on direct Ci/Gli regulation. All but one (orthodenticle) contain more than one Ci/Gli site, prompting us to directly test whether homotypic clustering of Ci/Gli binding sites is sufficient to define a Hh-regulated enhancer. We therefore developed a computational algorithm to identify Ci/Gli clusters that are enriched over random expectation, within a given region of the genome. Candidate genomic regions containing Ci/Gli clusters were functionally tested in chicken neural tube electroporation assays and in transgenic flies. Of the 22 Ci/Gli clusters tested, seven novel enhancers (and the previously known patched enhancer) were identified as Hh-responsive and Ci/Gli-dependent in one or both of these assays, including: Cuticular protein 100A (Cpr100A); invected (inv), which encodes an engrailed-related transcription factor expressed at the anterior/posterior wing disc boundary; roadkill (rdx), the fly homolog of vertebrate Spop; the segment polarity gene gooseberry (gsb); and two previously untested regions of the Hh receptor-encoding patched (ptc) gene. We conclude that homotypic Ci/Gli clustering is not sufficient information to ensure Hh-responsiveness; however, it can provide a clue for enhancer recognition within putative Hedgehog target gene loci. PMID:26710299

  6. Regulation of Calvarial Osteogenesis by Concomitant De-repression of GLI3 and Activation of IHH Targets

    Lotta K. Veistinen

    2017-12-01

    Full Text Available Loss-of-function mutations in GLI3 and IHH cause craniosynostosis and reduced osteogenesis, respectively. In this study, we show that Ihh ligand, the receptor Ptch1 and Gli transcription factors are differentially expressed in embryonic mouse calvaria osteogenic condensations. We show that in both Ihh−/− and Gli3Xt−J/Xt−J embryonic mice, the normal gene expression architecture is lost and this results in disorganized calvarial bone development. RUNX2 is a master regulatory transcription factor controlling osteogenesis. In the absence of Gli3, RUNX2 isoform II and IHH are upregulated, and RUNX2 isoform I downregulated. This is consistent with the expanded and aberrant osteogenesis observed in Gli3Xt−J/Xt−J mice, and consistent with Runx2-I expression by relatively immature osteoprogenitors. Ihh−/− mice exhibited small calvarial bones and HH target genes, Ptch1 and Gli1, were absent. This indicates that IHH is the functional HH ligand, and that it is not compensated by another HH ligand. To decipher the roles and potential interaction of Gli3 and Ihh, we generated Ihh−/−;Gli3Xt−J/Xt−J compound mutant mice. Even in the absence of Ihh, Gli3 deletion was sufficient to induce aberrant precocious ossification across the developing suture, indicating that the craniosynostosis phenotype of Gli3Xt−J/Xt−J mice is not dependent on IHH ligand. Also, we found that Ihh was not required for Runx2 expression as the expression of RUNX2 target genes was unaffected by deletion of Ihh. To test whether RUNX2 has a role upstream of IHH, we performed RUNX2 siRNA knock down experiments in WT calvarial osteoblasts and explants and found that Ihh expression is suppressed. Our results show that IHH is the functional HH ligand in the embryonic mouse calvaria osteogenic condensations, where it regulates the progression of osteoblastic differentiation. As GLI3 represses the expression of Runx2-II and Ihh, and also elevates the Runx2-I expression

  7. Regulation of Calvarial Osteogenesis by Concomitant De-repression of GLI3 and Activation of IHH Targets.

    Veistinen, Lotta K; Mustonen, Tuija; Hasan, Md Rakibul; Takatalo, Maarit; Kobayashi, Yukiho; Kesper, Dörthe A; Vortkamp, Andrea; Rice, David P

    2017-01-01

    Loss-of-function mutations in GLI3 and IHH cause craniosynostosis and reduced osteogenesis, respectively. In this study, we show that Ihh ligand, the receptor Ptch1 and Gli transcription factors are differentially expressed in embryonic mouse calvaria osteogenic condensations. We show that in both Ihh -/- and Gli3 Xt - J / Xt - J embryonic mice, the normal gene expression architecture is lost and this results in disorganized calvarial bone development. RUNX2 is a master regulatory transcription factor controlling osteogenesis. In the absence of Gli3 , RUNX2 isoform II and IHH are upregulated, and RUNX2 isoform I downregulated. This is consistent with the expanded and aberrant osteogenesis observed in Gli3 Xt - J / Xt - J mice, and consistent with Runx2-I expression by relatively immature osteoprogenitors. Ihh -/- mice exhibited small calvarial bones and HH target genes, Ptch1 and Gli1 , were absent. This indicates that IHH is the functional HH ligand, and that it is not compensated by another HH ligand. To decipher the roles and potential interaction of Gli3 and Ihh, we generated Ihh -/- ; Gli3 Xt - J / Xt - J compound mutant mice. Even in the absence of Ihh, Gli3 deletion was sufficient to induce aberrant precocious ossification across the developing suture, indicating that the craniosynostosis phenotype of Gli3 Xt - J / Xt - J mice is not dependent on IHH ligand. Also, we found that Ihh was not required for Runx2 expression as the expression of RUNX2 target genes was unaffected by deletion of Ihh . To test whether RUNX2 has a role upstream of IHH, we performed RUNX2 siRNA knock down experiments in WT calvarial osteoblasts and explants and found that Ihh expression is suppressed. Our results show that IHH is the functional HH ligand in the embryonic mouse calvaria osteogenic condensations, where it regulates the progression of osteoblastic differentiation. As GLI3 represses the expression of Runx2-II and Ihh , and also elevates the Runx2-I expression, and as IHH

  8. Phe-Gly dipeptidomimetics designed for the di-/tripeptide transporters PEPT1 and PEPT2

    Våbenø, Jon; Lejon, Tore; Nielsen, Carsten Uhd

    2004-01-01

    A series of five Phe-Gly dipeptidomimetics containing different amide bond replacements have been synthesized in a facile way from the readily available unsaturated ketoester 1, and their affinities for the di-/tripeptide transporters hPEPT1 (Caco-2 cells) and rPEPT2 (SKPT cells) were tested...... information about the importance of flexibility and of the stereochemistry at the C(4)-position for this class of compounds. Furthermore, the intracellular uptake of 2a-4a in Caco-2 cells was investigated, showing a 3-fold reduction of the uptake of 2a in the presence of the competetive inhibitor Gly......-Pro, indicating contribution from an active transport component. No active uptake of 3a and 4a was observed. Transepithelial transport studies also indicated active transport of 2a across Caco-2 monolayers....

  9. Surface Chirality of Gly-Pro Dipeptide Adsorbed on a Cu(110) Surface.

    Cruguel, Hervé; Méthivier, Christophe; Pradier, Claire-Marie; Humblot, Vincent

    2015-07-01

    The adsorption of chiral Gly-Pro dipeptide on Cu(110) has been characterized by combining in situ polarization modulation infrared reflection absorption spectroscopy (PM-RAIRS) and X-ray photoelectron spectroscopy (XPS). The chemical state of the dipeptide, and its anchoring points and adsorption geometry, were determined at various coverage values. Gly-Pro molecules are present on Cu(110) in their anionic form (NH2 /COO(-)) and adsorb under a 3-point binding via both oxygen atoms of the carboxylate group and via the nitrogen atom of the amine group. Low-energy electron diffraction (LEED) and scanning tunneling microscopy (STM) have shown the presence of an extended 2D chiral array, sustained via intermolecular H-bonds interactions. Furthermore, due to the particular shape of the molecule, only one homochiral domain is formed, creating thus a truly chiral surface. © 2015 Wiley Periodicals, Inc.

  10. Characterization of a TK6-Bcl-xL gly-159-ala Human Lymphoblast Clone

    Chyall, L.: Gauny, S.; Kronenberg, A.

    2006-01-01

    TK6 cells are a well-characterized human B-lymphoblast cell line derived from WIL-2 cells. A derivative of the TK6 cell line that was stably transfected to express a mutated form of the anti-apoptotic protein Bcl-xL (TK6-Bcl-xL gly-159- ala clone #38) is compared with the parent cell line. Four parameters were evaluated for each cell line: growth under normal conditions, plating efficiency, and frequency of spontaneous mutation to 6‑thioguanine resistance (hypoxanthine phosphoribosyl transferase locus) or trifluorothymidine resistance (thymidine kinase locus). We conclude that the mutated Bcl-xL protein did not affect growth under normal conditions, plating efficiency or spontaneous mutation frequencies at the thymidine kinase (TK) locus. Results at the hypoxanthine phosphoribosyl transferase (HPRT) locus were inconclusive. A mutant fraction for TK6‑Bcl-xL gly-159-ala clone #38 cells exposed to 150cGy of 160kVp x-rays was also calculated. Exposure to x-irradiation increased the mutant fraction of TK6‑Bcl-xL gly-159-ala clone #38 cells.

  11. Assessment of airborne soy-hull allergen (Gly m 1) in the Port of Ancona, Italy.

    Antonicelli, L; Ruello, M L; Monsalve, R I; González, R; Fava, G; Bonifazi, F

    2010-10-01

    Epidemic asthma outbreaks are potentially a very high-risk medical situation in seaport towns where large volumes of soybean are loaded and unloaded Airborne allergen assessment plays a pivotal role in evaluating the resulting environmental pollution. The aim of this study was to measure the airborne Gly m 1 allergen level in the seaport of Ancona in order assess the soybean-specific allergenic risk for the city. Allergen and PM10 were evaluated at progressive distances from the port area. Allergen analysis was performed by monoclonal antibody-based immunoassay on the sampled filters. Daily meteorological data were obtained from the local meteorological station. For estimating the assimilative capacity of the atmosphere, an approach based on dispersive ventilation coefficient was tried. The allergen concentrations detected were low (range = 0.4-171 ng/m3). A decreasing gradient of the airborne allergen from the unloading area (22.1 +/- 41.2 ng/m3) to the control area (0.6 +/- 0.7 ng/m3) was detected. The concentration of the airborne Gly m 1 was not coupled with the presence of the soy-carrying ships in the port. A statistically significant relationship between airborne allergen, PM10 and local meteorological parameters quantifies the association with the atmospheric condition. Airborne Gly m 1 is part of the atmospheric dust of Ancona. The low level of this allergen seems consistent with the absence of asthma epidemic outbreak.

  12. MicroRNAs Promote Granule Cell Expansion in the Cerebellum Through Gli2.

    Constantin, Lena; Wainwright, Brandon J

    2015-12-01

    MicroRNAs (miRNAs) are important regulators of cerebellar function and homeostasis. Their deregulation results in cerebellar neuronal degeneration and spinocerebellar ataxia type 1 and contributes to medulloblastoma. Canonical miRNA processing involves Dicer, which cleaves precursor miRNAs into mature double-stranded RNA duplexes. In order to address the role of miRNAs in cerebellar granule cell precursor development, loxP-flanked exons of Dicer1 were conditionally inactivated using the granule cell precursor-specific Atoh1-Cre recombinase. A reduction of 87% in Dicer1 transcript was achieved in this conditional Dicer knockdown model. Although knockdown resulted in normal survival, mice had disruptions to the cortical layering of the anterior cerebellum, which resulted from the premature differentiation of granule cell precursors in this region during neonatal development. This defect manifested as a thinner external granular layer with ectopic mature granule cells, and a depleted internal granular layer. We found that expression of the activator components of the Hedgehog-Patched pathway, the Gli family of transcription factors, was perturbed in conditional Dicer knockdown mice. We propose that loss of Gli2 mRNA mediated the anterior-restricted defect in conditional Dicer knockdown mice and, as proof of principle, were able to show that miR-106b positively regulated Gli2 mRNA expression. These findings confirm the importance of miRNAs as positive mediators of Hedgehog-Patched signalling during granule cell precursor development.

  13. Relationship between ADD1 Gly460Trp gene polymorphism and essential hypertension in Madeira Island.

    Sousa, Ana Célia; Palma Dos Reis, Roberto; Pereira, Andreia; Borges, Sofia; Freitas, Ana Isabel; Guerra, Graça; Góis, Teresa; Rodrigues, Mariana; Henriques, Eva; Freitas, Sónia; Ornelas, Ilídio; Pereira, Décio; Brehm, António; Mendonça, Maria Isabel

    2017-10-01

    Essential hypertension (EH) is a complex disease in which physiological, environmental, and genetic factors are involved in its genesis. The genetic variant of the alpha-adducin gene (ADD1) has been described as a risk factor for EH, but with controversial results.The objective of this study was to evaluate the association of ADD1 (Gly460Trp) gene polymorphism with the EH risk in a population from Madeira Island.A case-control study with 1614 individuals of Caucasian origin was performed, including 817 individuals with EH and 797 controls. Cases and controls were matched for sex and age, by frequency-matching method. All participants collected blood for biochemical and genotypic analysis for the Gly460Trp polymorphism. We further investigated which variables were independently associated to EH, and, consequently, analyzed their interactions.In our study, we found a significant association between the ADD1 gene polymorphism and EH (odds ratio 2.484, P = .01). This association remained statistically significant after the multivariate analysis (odds ratio 2.548, P = .02).The ADD1 Gly460Trp gene polymorphism is significantly and independently associated with EH risk in our population. The knowledge of genetic polymorphisms associated with EH is of paramount importance because it leads to a better understanding of the etiology and pathophysiology of this pathology.

  14. Synergism between Hedgehog-GLI and EGFR signaling in Hedgehog-responsive human medulloblastoma cells induces downregulation of canonical Hedgehog-target genes and stabilized expression of GLI1.

    Frank Götschel

    Full Text Available Aberrant activation of Hedgehog (HH signaling has been identified as a key etiologic factor in many human malignancies. Signal strength, target gene specificity, and oncogenic activity of HH signaling depend profoundly on interactions with other pathways, such as epidermal growth factor receptor-mediated signaling, which has been shown to cooperate with HH/GLI in basal cell carcinoma and pancreatic cancer. Our experimental data demonstrated that the Daoy human medulloblastoma cell line possesses a fully inducible endogenous HH pathway. Treatment of Daoy cells with Sonic HH or Smoothened agonist induced expression of GLI1 protein and simultaneously prevented the processing of GLI3 to its repressor form. To study interactions between HH- and EGF-induced signaling in greater detail, time-resolved measurements were carried out and analyzed at the transcriptomic and proteomic levels. The Daoy cells responded to the HH/EGF co-treatment by downregulating GLI1, PTCH, and HHIP at the transcript level; this was also observed when Amphiregulin (AREG was used instead of EGF. We identified a novel crosstalk mechanism whereby EGFR signaling silences proteins acting as negative regulators of HH signaling, as AKT- and ERK-signaling independent process. EGFR/HH signaling maintained high GLI1 protein levels which contrasted the GLI1 downregulation on the transcript level. Conversely, a high-level synergism was also observed, due to a strong and significant upregulation of numerous canonical EGF-targets with putative tumor-promoting properties such as MMP7, VEGFA, and IL-8. In conclusion, synergistic effects between EGFR and HH signaling can selectively induce a switch from a canonical HH/GLI profile to a modulated specific target gene profile. This suggests that there are more wide-spread, yet context-dependent interactions, between HH/GLI and growth factor receptor signaling in human malignancies.

  15. Msx genes are important apoptosis effectors downstream of the Shh/Gli3 pathway in the limb.

    Lallemand, Yvan; Bensoussan, Vardina; Cloment, Cécile Saint; Robert, Benoît

    2009-07-15

    In tetrapods, the anteroposterior (AP) patterning of the limb is under the control of the antagonistic activities of the secreted factor Sonic hedgehog (Shh) and Gli3R, the truncated repressor form of the transcription factor Gli3. In this report, we show that Msx1 and Msx2 are targets and downstream effectors of Gli3R. Consequently, in Shh null mutants, Msx genes are overexpressed and, furthermore, partially responsible for the limb phenotype. This is exemplified by the fact that reducing Msx activity in Shh mutants partially restores a normal limb development. Finally, we show that the main action of the Msx genes, in both normal and Shh(-/-) limb development, is to control cell death in the mesenchyme. We propose that, in the limb, Msx genes act downstream of the Shh/Gli3 pathway by transducing BMP signaling and that, in the absence of Shh signaling, their deregulation contributes to the extensive apoptosis that impairs limb development.

  16. AMP-Activated Protein Kinase Directly Phosphorylates and Destabilizes Hedgehog Pathway Transcription Factor GLI1 in Medulloblastoma

    Yen-Hsing Li

    2015-07-01

    Full Text Available The Hedgehog (Hh pathway regulates cell differentiation and proliferation during development by controlling the Gli transcription factors. Cell fate decisions and progression toward organ and tissue maturity must be coordinated, and how an energy sensor regulates the Hh pathway is not clear. AMP-activated protein kinase (AMPK is an important sensor of energy stores and controls protein synthesis and other energy-intensive processes. AMPK is directly responsive to intracellular AMP levels, inhibiting a wide range of cell activities if ATP is low and AMP is high. Thus, AMPK can affect development by influencing protein synthesis and other processes needed for growth and differentiation. Activation of AMPK reduces GLI1 protein levels and stability, thus blocking Sonic-hedgehog-induced transcriptional activity. AMPK phosphorylates GLI1 at serines 102 and 408 and threonine 1074. Mutation of these three sites into alanine prevents phosphorylation by AMPK. This leads to increased GLI1 protein stability, transcriptional activity, and oncogenic potency.

  17. Lithium Suppresses Hedgehog Signaling via Promoting ITCH E3 Ligase Activity and Gli1–SUFU Interaction in PDA Cells

    Xinshuo Wang

    2017-11-01

    Full Text Available Dysregulation of Hedgehog (Hh signaling pathway is one of the hallmarks of pancreatic ductal adenocarcinoma (PDA. Lithium, a clinical mood stabilizer for the treatment of mental disorders, is known to suppress tumorigenic potential of PDA cells by targeting the Hh/Gli signaling pathway. In this study, we investigated the molecular mechanism of lithium induced down-regulation of Hh/Gli1. Our data show that lithium promotes the poly-ubiquitination and proteasome-mediated degradation of Gli1 through activating E3 ligase ITCH. Additionally, lithium enhances interaction between Gli1 and SUFU via suppressing GSK3β, which phosphorylates SUFU and destabilizes the SUFU-Gli1 inhibitory complex. Our studies illustrate a novel mechanism by which lithium suppresses Hh signaling via simultaneously promoting ITCH-dependent Gli1 ubiquitination/degradation and SUFU-mediated Gli1 inhibition.

  18. Book review. Conoscere gli animali familiari. Francesca Bellini, Alessia Liverini, Vincenzo Rosa

    Manuel Graziani

    2014-03-01

    Full Text Available Conoscere gli animali familiari è la settima uscita della collana diretta da Paolo Polidori "produzioni animali e sicurezza alimentare" sulla ricerca nell'ambito della nutrizione e alimentazione animale, zootecnia, ispezione degli alimenti di origine animale, clinica medica e parassitologia veterinaria con risvolti di natura tecnica, scientifica e pratica. Il volume, redatto da medici veterinari in servizio presso differenti Aziende Sanitarie italiane, è un sintetico manuale su domesticazione, cura sanitaria, etnografia e addestramento del proprio cane (o gatto. Gli autori partono dall'assunto che i proprietari scelgono il cane per lo più sulla base del gusto visivo o sulla consuetudine tramandata in famiglia, senza un'esaustiva conoscenza degli aspetti riguardanti l'origine dell'animale, la sua attitudine prevalente, le sue patologie ricorrenti e le eventuali predisposizioni genetiche nei confronti di una determinata patologia. Puntano l'accento sin dalle prime battute sul cambiamento culturale che ha modificato anche il rapporto uomo-animale sotto l'aspetto sociale, effettuale e giuridico. L'animale da semplice "res" si è da tempo affermato come un essere diverso ma senziente, quindi destinatario di tutele, con diritti contemplati dalle carte costituzionali di diversi Paesi. Per questo motivo chi detiene un animale domestico deve prepararsi ad un impegno non riducibile al possesso di un oggetto, al punto di risponderne penalmente per eventuali sofferenze fisiche o psicologiche, per l'abbandono (anche solo temporaneo o per l'incuria. Conoscere gli animali familiari fornisce nozioni scientifiche e sanitarie alla portata di tutti, utili soprattutto nelle occasioni in cui i proprietari dovranno recarsi dal veterinario o affrontare visite legate alla profilassi e alle vaccinazioni di routine.

  19. Gli infortuni lavorativi in minori: risultati di uno studio multicentrico italiano

    G. Aggazzotti

    2003-05-01

    Full Text Available

    Obiettivi: gli infortuni lavorativi, in particolare quelli subiti da minorenni, rappresentano un problema di notevole importanza sociale: in Italia, da dati INPS, risultano lavorare circa 68 adolescenti su 1000. L’obiettivo di questo studio è stato quello di raccogliere informazioni sugli infortuni lavorativi avvenuti a minori (14-17 anni in 14 città italiane nel periodo gennaio-giugno 2000. Metodi: si tratta di uno studio epidemiologico descrittivo, basato sulle informazioni raccolte consultando direttamente le cartelle cliniche presso centri di Pronto Soccorso (PS attivi nelle città coinvolte. Sono stati presi in considerazione tutti gli infortuni avvenuti a minori: tra questi sono stati considerati come lavorativi quelli accaduti nel corso di attività descritta come lavorativa e quelli occorsi “in itinere”. Le analisi statistiche sono state condotte con SPSS; è stata effettuata una cluster analysis per evidenziare eventuali sottogruppi omogenei. Risultati: la popolazione residente nelle aree indagate di età tra 14 e 17 anni è stata stimata in circa 850.000 soggetti, pari al 31% della popolazione italiana della stessa età: gli infortuni totali sono risultati 13423 di cui 317 lavorativi (2.4%. I soggetti di sesso maschile, diciassettenni, impiegati nel comparto industriale sono risultati il gruppo maggiormente coinvolto: la prognosi è risultata per lo più inferiore a 8 giorni. Sono apparse due diverse tipologie di infortunio: una riguarda i casi avvenuti in itinere, con caratteristiche molto simili agli incidenti stradali, e un’altra, più specifica, riguarda gli eventi sul posto di lavoro, dove si registrano lesioni al polso, alle mani, al capo e agli occhi con frequenza superiore rispetto agli infortuni in genere.

    Conclusioni: il fenomeno è risultato non trascurabile, soprattutto tenendo conto del fatto che si riferisce solamente al lavoro minorile regolarmente

  20. Structure/Function Studies of the α4 Subunit Reveal Evolutionary Loss of a GlyR Subtype Involved in Startle and Escape Responses

    Sophie Leacock

    2018-01-01

    Full Text Available Inhibitory glycine receptors (GlyRs are pentameric ligand-gated anion channels with major roles in startle disease/hyperekplexia (GlyR α1, cortical neuronal migration/autism spectrum disorder (GlyR α2, and inflammatory pain sensitization/rhythmic breathing (GlyR α3. However, the role of the GlyR α4 subunit has remained enigmatic, because the corresponding human gene (GLRA4 is thought to be a pseudogene due to an in-frame stop codon at position 390 within the fourth membrane-spanning domain (M4. Despite this, a recent genetic study has implicated GLRA4 in intellectual disability, behavioral problems and craniofacial anomalies. Analyzing data from sequenced genomes, we found that GlyR α4 subunit genes are predicted to be intact and functional in the majority of vertebrate species—with the exception of humans. Cloning of human GlyR α4 cDNAs excluded alternative splicing and RNA editing as mechanisms for restoring a full-length GlyR α4 subunit. Moreover, artificial restoration of the missing conserved arginine (R390 in the human cDNA was not sufficient to restore GlyR α4 function. Further bioinformatic and mutagenesis analysis revealed an additional damaging substitution at K59 that ablates human GlyR α4 function, which is not present in other vertebrate GlyR α4 sequences. The substitutions K59 and X390 were also present in the genome of an ancient Denisovan individual, indicating that GLRA4 has been a pseudogene for at least 30,000–50,000 years. In artificial synapses, we found that both mouse and gorilla α4β GlyRs mediate synaptic currents with unusually slow decay kinetics. Lastly, to gain insights into the biological role of GlyR α4 function, we studied the duplicated genes glra4a and glra4b in zebrafish. While glra4b expression is restricted to the retina, using a novel tol2-GAL4FF gene trap line (SAIGFF16B, we found that the zebrafish GlyR α4a subunit gene (glra4a is strongly expressed in spinal cord and hindbrain commissural

  1. Reactivity of cosmetic UV filters towards skin proteins: model studies with Boc-lysine, Boc-Gly-Phe-Gly-Lys-OH, BSA and gelatin.

    Stiefel, C; Schwack, W

    2014-12-01

    Organic UV filters are used as active ingredients in most sunscreens and also in a variety of daily care products. Their good (photo) stability is of special interest to guarantee protective function and to prevent interactions with the human skin. Due to the mostly electrophilic character of the UV filters, reactions with nucleophilic protein moieties like lysine side chains are conceivable. Prior studies showed that the UV filters octocrylene (OCR), butyl methoxydibenzoylmethane (BM-DBM), ethylhexyl salicylate (EHS), ethylhexyl methoxycinnamate (EHMC), benzophenone-3 (BP-3), ethylhexyl triazone (EHT) and dibenzoylmethane (DBM) were able to covalently bind to an HPTLC amino phase and the amino acid models ethanolamine and butylamine after slightly heating and/or radiation. Boc-protected lysine, the tetrapeptide Boc-Gly-Phe-Gly-Lys-OH, bovine serum albumin (BSA) and porcine gelatin were used as more complex models to determine the reactivity of the mentioned UV filters towards skin proteins under thermal or UV irradiation conditions. After gentle heating at 37°C, benzophenone imines were identified as reaction products of BP-3 and OCR with Boc-lysine and the tetrapeptide, whereas DBM and BM-DBM yielded enamines. For EHMC, a Michael-type reaction occurred, which resulted in addition of Boc-lysine or the tetrapeptide to the conjugated double bond. Ester aminolysis of EHS and EHT mainly afforded the corresponding amides. Reactions of the UV filters with BSA changed the UV spectrum of BSA, generally associated with an increase of the absorption strength in the UVA or UVB range. For all protein models, the UV filters showed an increasing reactivity in the order EHT < EHMC < EHS < BP-3 < OCR < DBM < BM-DBM. Especially the UV absorbers BM-DBM, OCR and BP-3, which are seen as common allergens or photoallergens, showed a high reactivity towards the different skin protein models. As the formation of protein adducts is recognized as important key element in the induction of

  2. Molecular shape and binding force of Mycoplasma mobile's leg protein Gli349 revealed by an AFM study

    Lesoil, Charles [Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsutacho 4259, Midori-ku, Yokohama 226-8501 (Japan); Nonaka, Takahiro [Department of Biology, Graduate School of Science, Osaka City University, Sumiyoshi-ku, Osaka 558-8585 (Japan); Sekiguchi, Hiroshi; Osada, Toshiya [Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsutacho 4259, Midori-ku, Yokohama 226-8501 (Japan); Miyata, Makoto [Department of Biology, Graduate School of Science, Osaka City University, Sumiyoshi-ku, Osaka 558-8585 (Japan); Afrin, Rehana [Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsutacho 4259, Midori-ku, Yokohama 226-8501 (Japan); Biofrontier Center, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsutacho 4259, Midori-ku, Yokohama 226-8501 (Japan); Ikai, Atsushi, E-mail: ikai.a.aa@m.titech.ac.jp [Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsutacho 4259, Midori-ku, Yokohama 226-8501 (Japan)

    2010-01-15

    Recent studies of the gliding bacteria Mycoplasma mobile have identified a family of proteins called the Gli family which was considered to be involved in this novel and yet fairly unknown motility system. The 349 kDa protein called Gli349 was successfully isolated and purified from the bacteria, and electron microscopy imaging and antibody experiments led to the hypothesis that it acts as the 'leg' of M. mobile, responsible for attachment to the substrate as well as for gliding motility. However, more precise evidence of the molecular shape and function of this protein was required to asses this theory any further. In this study, an atomic force microscope (AFM) was used both as an imaging and a force measurement device to provide new information about Gli349 and its role in gliding motility. AFM images of the protein were obtained revealing a complex structure with both rigid and flexible parts, consistent with previous electron micrographs of the protein. Single-molecular force spectroscopy experiments were also performed, revealing that Gli349 is able to specifically bind to sialyllactose molecules and withstand unbinding forces around 70 pN. These findings strongly support the idea that Gli349 is the 'leg' protein of M. mobile, responsible for binding and also most probably force generation during gliding motility.

  3. Molecular shape and binding force of Mycoplasma mobile's leg protein Gli349 revealed by an AFM study

    Lesoil, Charles; Nonaka, Takahiro; Sekiguchi, Hiroshi; Osada, Toshiya; Miyata, Makoto; Afrin, Rehana; Ikai, Atsushi

    2010-01-01

    Recent studies of the gliding bacteria Mycoplasma mobile have identified a family of proteins called the Gli family which was considered to be involved in this novel and yet fairly unknown motility system. The 349 kDa protein called Gli349 was successfully isolated and purified from the bacteria, and electron microscopy imaging and antibody experiments led to the hypothesis that it acts as the 'leg' of M. mobile, responsible for attachment to the substrate as well as for gliding motility. However, more precise evidence of the molecular shape and function of this protein was required to asses this theory any further. In this study, an atomic force microscope (AFM) was used both as an imaging and a force measurement device to provide new information about Gli349 and its role in gliding motility. AFM images of the protein were obtained revealing a complex structure with both rigid and flexible parts, consistent with previous electron micrographs of the protein. Single-molecular force spectroscopy experiments were also performed, revealing that Gli349 is able to specifically bind to sialyllactose molecules and withstand unbinding forces around 70 pN. These findings strongly support the idea that Gli349 is the 'leg' protein of M. mobile, responsible for binding and also most probably force generation during gliding motility.

  4. GLI1, a crucial mediator of sonic hedgehog signaling in prostate cancer, functions as a negative modulator for androgen receptor

    Chen, Guangchun; Goto, Yutaka; Sakamoto, Ryuichi; Tanaka, Kimitaka; Matsubara, Eri; Nakamura, Masafumi; Zheng, Hong; Lu, Jian; Takayanagi, Ryoichi; Nomura, Masatoshi

    2011-01-01

    Research highlights: → GLI1, which play a central role in sonic hedgehog signaling in prostate cancer, can act as a co-repressor to substantially block androgen receptor-mediated transactivation. → GLI1 directly interacts with AR. → SHH-GLI pathway might be one of determinants governing the transition of prostate cancer from an androgen-dependent to an androgen-independent state. -- Abstract: Sonic hedgehog (SHH) signaling, acting in a combinatorial manner with androgen signaling, is essential for prostate patterning and development. Recently, elevated activation of SHH signaling has been shown to play important roles in proliferation, progression and metastasis of prostate cancer. In this report, we demonstrate for the first time, that GLI1, which has been shown to play a central role in SHH signaling in prostate cancer, can act as a co-repressor to substantially block androgen receptor (AR)-mediated transactivation, at least in part, by directly interacting with AR. Our observations suggest that the SHH-GLI pathway might be one of determinants governing the transition of prostate cancer from an androgen-dependent to an androgen-independent state by compensating, or even superseding androgen signaling.

  5. Constitutive activation of Gli2 impairs bone formation in postnatal growing mice.

    Kyu Sang Joeng

    Full Text Available Indian hedgehog (Ihh signaling is indispensable for osteoblast differentiation during endochondral bone development in the mouse embryo. We have previously shown that the Gli2 transcription activator critically mediates Ihh function in osteoblastogenesis. To explore the possibility that activation of Hedgehog (Hh signaling may enhance bone formation, we generated mice that expressed a constitutively active form of Gli2 in the Osx-lineage cells. Unexpectedly, these mice exhibited severe osteopenia due to a marked decrease in osteoblast number and function, although bone resorption was not affected. Quantitative analyses of the molecular markers indicated that osteoblast differentiation was impaired in the mutant mouse. However, the osteoblast-lineage cells isolated from these mice exhibited more robust osteoblast differentiation than normal in vitro. Similarly, pharmacological stimulation of Hh signaling enhanced osteoblast differentiation from Osx-expressing cells isolated from the wild-type mouse. Thus, even though Hh signaling directly promotes osteoblast differentiation in vitro, constitutive activation of this pathway impairs bone formation in vivo, perhaps through an indirect mechanism.

  6. Molecular Determinants for Substrate Interactions with the Glycine Transporter GlyT2.

    Carland, Jane E; Thomas, Michael; Mostyn, Shannon N; Subramanian, Nandhitha; O'Mara, Megan L; Ryan, Renae M; Vandenberg, Robert J

    2018-03-21

    Transporters in the SLC6 family play key roles in regulating neurotransmission and are the targets for a wide range of therapeutics. Important insights into the transport mechanisms and the specificity of drug interactions of SLC6 transporters have been obtained from the crystal structures of a bacterial homologue of the family, LeuT Aa , and more recently the Drosophila dopamine transporter and the human serotonin transporter. However, there is disputed evidence that the bacterial leucine transporter, LeuT Aa , contains two substrate binding sites that work cooperatively in the mechanism of transport, with the binding of a second substrate being required for the release of the substrate from the primary site. An alternate proposal is that there may be low affinity binding sites that serve to direct the flow of substrates to the primary site. We have used a combination of molecular dynamics simulations of substrate interactions with a homology model of GlyT2, together with radiolabeled amino acid uptake assays and electrophysiological analysis of wild-type and mutant transporters, to provide evidence that substrate selectivity of GlyT2 is determined entirely by the primary substrate binding site and, furthermore, if a secondary site exists then it is a low affinity nonselective amino acid binding site.

  7. [CONTATTI SONORI] Morfologia e poetica della musica di Dave Holland fra gli anni Settanta e Ottanta

    Giuseppe Sergi

    2015-02-01

    Full Text Available La musica del contrabbassista inglese Dave Holland ha radici nel jazz ma dialoga con i linguaggi della tradizione colta. Le esigenze compositive si affiancano alla necessità dell'improvvisazione. Holland espone così la propria poetica: «gli uccelli si riuniscono per cantare insieme, ognuno affermando con il canto la propria libertà. Il mio desiderio è condividere il medesimo spirito con i musicisti e comunicarlo alla gente». Questo studio affronta un'analisi di incisioni significative che mettono in luce i risultati musicali di tale pensiero. Il brano Conference of the birds è un manifesto poetico: esso spicca per le scelte sonore, la metrica, l'incrocio delle voci e le giustapposizioni modali. First snow si caratterizza per la correlazione tra il trattamento armonico e la natura modale delle proposte melodiche. Homecoming presenta una peculiare sequenza di episodi musicali e un significativo rapporto fra soluzioni melodiche e armoniche di carattere tonale e modale. L'analisi solleva interrogativi di rilievo: quali strategie compositive e performative vengono adottate nella musica di Dave Holland? Che legame esse intrattengono con il linguaggio del jazz o della musica 'd'arte'? Che tipo di equilibrio si crea fra la dimensione compositiva e l'improvvisazione? Tentando di fornire risposte a tali domande questo studio mette in luce gli elementi rilevanti della musica di uno maggiori compositori e musicisti del jazz strumentale.

  8. Phenotypic heterogeneity associated with a novel mutation (Gly112Glu) in the Norrie disease protein.

    Allen, R C; Russell, S R; Streb, L M; Alsheikheh, A; Stone, E M

    2006-02-01

    To determine the molecular pathology and clinical severity of two pedigrees with a history of early retinal detachment and peripheral retinal vascular abnormalities. Longitudinal cohort study. A longitudinal clinical study and DNA analysis was performed on 49 family members of two pedigrees. Nine individuals were found to be hemizygous for a mutation at codon 112 (Gly112Glu) of the Norrie disease protein (NDP) in one pedigree. Significant phenotypic heterogeneity was found. The proband presented with a unilateral subtotal retinal detachment at the age of 3 years, and subsequently developed a slowly progressive tractional retinal detachment involving the macula in the contralateral eye at the age of 4 years. One individual had only mild peripheral retinal pigmentary changes with normal vision at the age of 79 years. The remaining seven individuals had varying degrees of peripheral retinal vascular abnormalities and anterior segment findings. Seven affected members of a second pedigree affected by a previously reported mutation, Arg74Cys, also demonstrated wide ocular phenotypic variation. A novel mutation (Gly112Glu), which represents the most carboxy located, NDP mutation reported, results in significant phenotypic heterogeneity. These data support the contention that the spectrum of ocular disease severity associated with these NDP mutations is broad. Use of terms that characterize this entity by phenotypic appearance, such as familial exudative vitreoretinopathy, do not adequately communicate the potential spectrum of severity of this disorder to affected or carrier family members.

  9. Loss-of-Function of Gli3 in Mice Causes Abnormal Frontal Bone Morphology and Premature Synostosis of the Interfrontal Suture

    Veistinen, Lotta; Takatalo, Maarit; Tanimoto, Yukiho; Kesper, Dörthe A.; Vortkamp, Andrea; Rice, David P. C.

    2012-01-01

    Greig cephalopolysyndactyly syndrome (GCPS) is an autosomal dominant disorder with polydactyly and syndactyly of the limbs and a broad spectrum of craniofacial abnormalities. Craniosynostosis of the metopic suture (interfrontal suture in mice) is an important but rare feature associated with GCPS. GCPS is caused by mutations in the transcription factor GLI3, which regulates Hedgehog signaling. The Gli3 loss-of-function (Gli3Xt-J/Xt-J) mouse largely phenocopies the human syndrome with the mice...

  10. Synthesis and structure-activity relationship of N-alkyl Gly-boro-Pro inhibitors of DPP4, FAP, and DPP7.

    Hu, Yi; Ma, Lifu; Wu, Min; Wong, Melissa S; Li, Bei; Corral, Sergio; Yu, Zhizhou; Nomanbhoy, Tyzoon; Alemayehu, Senaiet; Fuller, Stacy R; Rosenblum, Jonathan S; Rozenkrants, Natasha; Minimo, Lauro C; Ripka, William C; Szardenings, Anna K; Kozarich, John W; Shreder, Kevin R

    2005-10-01

    The structure-activity relationship of various N-alkyl Gly-boro-Pro derivatives against three dipeptidyl peptidases (DPPs) was studied. In a series of N-cycloalkyl analogs, DPP4 and fibroblast activation protein-alpha (FAP) optimally preferred N-cycloheptyl whereas DPP7 tolerated even larger cycloalkyl rings. Gly alpha-carbon derivatization of N-cyclohexyl or N-(2-adamantyl) Gly-boro-Pro resulted in a significant decrease in potency against all the three DPPs.

  11. Design and Synthesis of Novel and Selective Glycine Transporter-1 (GlyT1) Inhibitors with Memory Enhancing Properties.

    Santora, Vincent J; Almos, Theresa A; Barido, Richard; Basinger, Jillian; Bellows, Chris L; Bookser, Brett Carder; Breitenbucher, J Guy; Broadbent, Nicola J; Cabebe, Clifford; Chai, Chih-Kun; Chen, Mi; Chow, Stephine; Chung, De Michael; Crickard, Lindsay; Danks, Anne M; Freestone, Graeme; Gitnick, Dany; Gupta, Varsha; Hoffmaster, Christine; Hudson, Andrew R; Kaplan, Alan P; Kennedy, Michael R; Lee, Dong; Limberis, James; Ly, Kiev; Mak, Chi Ching; Masatsugu, Brittany; Morse, Andrew C; Na, Jim; Neul, David; Nikpur, John; Peters, Marco; Petroski, Robert E; Renick, Joel; Sebring, Kristen; Sevidal, Samantha; Tabatabaei, Ali; Wen, Jenny; Yan, Yingzhuo; Yoder, Zachary W; Zook, Douglas

    2018-06-11

    We report here the identification and optimization of a novel series of potent GlyT1 inhibitors. A ligand design campaign that utilized known GlyT1 inhibitors as starting points led to the identification of a novel series of pyrrolo[3,4-c]pyrazoles amides (21-50) with good in vitro potency. Subsequent optimization of physicochemical and in vitro ADME properties produced several compounds with promising pharmacokinetic profiles. In vivo inhibition of GlyT1 was demonstrated for select compounds within this series by measuring the elevation of glycine in the cerebrospinal fluid (CSF) of rats after a single oral dosing of 10 mg/kg. Ultimately, an optimized lead, compound 46, demonstrated in vivo efficacy in a rat Novel Object Recognition (NOR) assay after oral dosing at 0.1, 1, and 3 mg/kg.

  12. The Role of the PGC1α Gly482Ser Polymorphism in Weight Gain due to Intensive Diabetes Therapy

    Samir S. Deeb

    2009-01-01

    Full Text Available The Diabetes Control and Complications Trial (DCCT involved intensive diabetes therapy of subjects with type 1 diabetes mellitus (T1DM for an average period of 6.5 years. A subset of these subjects gained excessive weight. We tested for association of polymorphisms in 8 candidate genes with the above trait. We found the Gly482Ser polymorphism in the peroxisome proliferator-activated receptor γ coactivator-1α (PGC1α to be significantly associated with weight gain in males (P=.0045 but not in females. The Ser allele was associated with greater weight gain than the Gly allele (P=.005. Subjects with a family history of type 2 diabetes mellitus (T2DM were more common among those who gained excessive weight. We conclude that T2DM and the Gly482Ser polymorphism in PGC1α contribute to the effect of intensive diabetes therapy on weight gain in males with T1DM.

  13. Sall4-Gli3 system in early limb progenitors is essential for the development of limb skeletal elements.

    Akiyama, Ryutaro; Kawakami, Hiroko; Wong, Julia; Oishi, Isao; Nishinakamura, Ryuichi; Kawakami, Yasuhiko

    2015-04-21

    Limb skeletal elements originate from the limb progenitor cells, which undergo expansion and patterning to develop each skeletal element. Posterior-distal skeletal elements, such as the ulna/fibula and posterior digits develop in a Sonic hedgehog (Shh)-dependent manner. However, it is poorly understood how anterior-proximal elements, such as the humerus/femur, the radius/tibia and the anterior digits, are developed. Here we show that the zinc finger factors Sall4 and Gli3 cooperate for proper development of the anterior-proximal skeletal elements and also function upstream of Shh-dependent posterior skeletal element development. Conditional inactivation of Sall4 in the mesoderm before limb outgrowth caused severe defects in the anterior-proximal skeletal elements in the hindlimb. We found that Gli3 expression is reduced in Sall4 mutant hindlimbs, but not in forelimbs. This reduction caused posteriorization of nascent hindlimb buds, which is correlated with a loss of anterior digits. In proximal development, Sall4 integrates Gli3 and the Plzf-Hox system, in addition to proliferative expansion of cells in the mesenchymal core of nascent hindlimb buds. Whereas forelimbs developed normally in Sall4 mutants, further genetic analysis identified that the Sall4-Gli3 system is a common regulator of the early limb progenitor cells in both forelimbs and hindlimbs. The Sall4-Gli3 system also functions upstream of the Shh-expressing ZPA and the Fgf8-expressing AER in fore- and hindlimbs. Therefore, our study identified a critical role of the Sall4-Gli3 system at the early steps of limb development for proper development of the appendicular skeletal elements.

  14. Antithrombotic Protective Effects of Arg-Pro-Gly-Pro Peptide during Emotional Stress Provoked by Forced Swimming Test in Rats.

    Grigor'eva, M E; Lyapina, L A

    2017-01-01

    Blood coagulation was enhanced and all factors (total, enzyme, and non-enzyme) of the fibrinolytic system were suppressed in rats in 60 min after forced swimming test. Argininecontaining tetrapeptide glyproline Arg-Pro-Gly-Pro administered prior to this test activated fibrinolysis and prevented hypercoagulation. Administration of this peptide in 5 min after swimming test also enhanced anticoagulant, fibrinolytic, and antithrombotic activity of the blood. Therefore, glyproline Arg-Pro-Gly-Pro exerted both preventive and curative effects on the hemostasis system and prevented enhancement of blood coagulation provoked by emotional stress modeled by forced swimming test.

  15. Preclinical evaluation of isostructural Tc-99m- and Re-188-folate-Gly-Gly-Cys-Glu for folate receptor-positive tumor targeting.

    Kim, Woo Hyoung; Kim, Chang Guhn; Kim, Myoung Hyoun; Kim, Dae-Weung; Park, Cho Rong; Park, Ji Yong; Lee, Yun-Sang; Youn, Hyewon; Kang, Keon Wook; Jeong, Jae Min; Chung, June-Key

    2016-06-01

    The purpose of the present study was to prepare isostructural Tc-99m- and Re-188-folate-Gly-Gly-Cys-Glu (folate-GGCE), and to evaluate the feasibility of their use for folate receptor (FR)-targeted molecular imaging and as theranostic agents in a mouse tumor model. Folate-GGCE was synthesized using solid-phase peptide synthesis and radiolabeled with Tc-99m or Re-188. Radiochemical characterization was performed by radio-high-performance liquid chromatography. The biodistribution of Tc-99m-folate-GGCE was studied, with or without co-injection of excess free folate, in mice bearing both FR-positive (KB cell) and FR-negative (HT1080 cell) tumors. Biodistribution of Re-188-folate-GGCE was studied in mice bearing KB tumors. Serial planar scintigraphy was performed in the dual tumor mouse model after intravenous injection of Tc-99m-folate-GGCE. Serial micro-single photon emission computed tomography/computed tomography (SPECT/CT) studies were performed, with or without co-injection of excess free folate, in the mouse tumor model after injection of Tc-99m-folate-GGCE or Re-188-folate-GGCE. The radiolabeling efficiency and radiochemical stability of Tc-99m- and Re-188-folate-GGCE were more than 95 % for up to 4 h after radiolabeling. Uptake of Tc-99m-folate-GGCE at 1, 2, and 4 h after injection in KB tumor was 16.4, 23.2, and 17.6 % injected dose per gram (%ID/g), respectively. This uptake was suppressed by 97.4 % when excess free folate was co-administered. Tumor:normal organ ratios at 4 h for blood, liver, lung, muscle, and kidney were 54.3, 25.2, 38.3, 97.8, and 0.3, respectively. Tumor uptake of Re-188-folate-GGCE at 2, 4, 8, and 16 h after injection was 17.4, 21.7, 24.1, and 15.6 %ID/g, respectively. Tumor:normal organ ratios at 8 h for blood, liver, lung, muscle, and kidney were 126.8, 21.9, 54.8, 80.3, and 0.4, respectively. KB tumors were clearly visualized at a high intensity using serial scintigraphy and micro-SPECT/CT in mice injected with Tc-99m- or Re

  16. Constitutive endocytosis and turnover of the neuronal glycine transporter GlyT2 is dependent on ubiquitination of a C-terminal lysine cluster.

    Jaime de Juan-Sanz

    Full Text Available Inhibitory glycinergic neurotransmission is terminated by sodium and chloride-dependent plasma membrane glycine transporters (GlyTs. The mainly glial glycine transporter GlyT1 is primarily responsible for the completion of inhibitory neurotransmission and the neuronal glycine transporter GlyT2 mediates the reuptake of the neurotransmitter that is used to refill synaptic vesicles in the terminal, a fundamental role in the physiology and pathology of glycinergic neurotransmission. Indeed, inhibitory glycinergic neurotransmission is modulated by the exocytosis and endocytosis of GlyT2. We previously reported that constitutive and Protein Kinase C (PKC-regulated endocytosis of GlyT2 is mediated by clathrin and that PKC accelerates GlyT2 endocytosis by increasing its ubiquitination. However, the role of ubiquitination in the constitutive endocytosis and turnover of this protein remains unexplored. Here, we show that ubiquitination of a C-terminus four lysine cluster of GlyT2 is required for constitutive endocytosis, sorting into the slow recycling pathway and turnover of the transporter. Ubiquitination negatively modulates the turnover of GlyT2, such that increased ubiquitination driven by PKC activation accelerates transporter degradation rate shortening its half-life while decreased ubiquitination increases transporter stability. Finally, ubiquitination of GlyT2 in neurons is highly responsive to the free pool of ubiquitin, suggesting that the deubiquitinating enzyme (DUB ubiquitin C-terminal hydrolase-L1 (UCHL1, as the major regulator of neuronal ubiquitin homeostasis, indirectly modulates the turnover of GlyT2. Our results contribute to the elucidation of the mechanisms underlying the dynamic trafficking of this important neuronal protein which has pathological relevance since mutations in the GlyT2 gene (SLC6A5 are the second most common cause of human hyperekplexia.

  17. Genetic variations and alternative splicing. The Glioma associated oncogene 1, GLI1.

    Peter eZaphiropoulos

    2012-07-01

    Full Text Available Alternative splicing is a post-transcriptional regulatory process that is attaining stronger recognition as a modulator of gene expression. Alternative splicing occurs when the primary RNA transcript is differentially processed into more than one mature RNAs. This is the result of a variable definition/inclusion of the exons, the sequences that are excised from the primary RNA to form the mature RNAs. Consequently, RNA expression can generate a collection of differentially spliced RNAs, which may distinctly influence subsequent biological events, such as protein synthesis or other biomolecular interactions. Still the mechanisms that control exon definition and exon inclusion are not fully clarified. This mini-review highlights advances in this field as well as the impact of single nucleotide polymorphisms in affecting splicing decisions. The Glioma associated oncogene 1, GLI1, is taken as an example in addressing the role of nucleotide substitutions for splicing regulation.

  18. Gli arcipelaghi and Il Figlio di Bakunìn: the narrator in film and novel

    Myriam Mereu

    2012-12-01

    Full Text Available Obiettivo del presente contributo è analizzare la struttura di due noti romanzi sardi, Il figlio di Bakunìn di Sergio Atzeni e Gli arcipelaghi di Maria Giacobbe, caratterizzati da una narrazione in prima persona, e dimostrare come il materiale narrativo è riorganizzato e riplasmato nelle trasposizioni cinematografiche, dirette rispettivamente da Gianfranco Cabiddu (1997 e da Giovanni Columbu (2001. La locuzione impiegata per riferirsi ai film è quella di traduzione intersemiotica, che riprende la terminologia del linguista Roman Jakobson. In questo modo, si vogliono mettere in evidenza l’intento dei registi di ricreare i fili narrativi dei romanzi e la volontà di riproporre, reintepretandoli, i punti di vista di ciascun personaggio. In un secondo momento, l’attenzione sarà rivolta alle scelte linguistiche adottate nei romanzi e nei film.

  19. TLR4 Asp299Gly polymorphism may be protective against chronic periodontitis.

    Sellers, R M; Payne, J B; Yu, F; LeVan, T D; Walker, C; Mikuls, T R

    2016-04-01

    Periodontitis results from interplay between genetic and environmental factors. Single nucleotide polymorphisms (SNPs) in the coding region of the toll-like receptor 4 gene (TLR4) may be associated with periodontitis, although previous studies have been inconclusive. Moreover, the interaction between environmental factors, such as cigarette smoking (a major risk factor for periodontitis), and Porphyromonas gingivalis (a major periodontal pathogen) with the TLR4 coding region Asp299Gly SNP (rs4986790; a SNP associated with lipopolysaccharide-mediated inflammatory responses in periodontitis), have been largely ignored in previous reports. Therefore, the objective of this study was to examine the association between TLR4 Asp299Gly (rs4986790) with alveolar bone height loss (ABHL) and periodontitis, accounting for interactions between this SNP with smoking and P. gingivalis prevalence. The CD14/-260 SNP (rs2569190) served as a control, as a recent meta-analysis suggested no relationship between this SNP and periodontitis. This multicenter study included 617 participants who had rheumatoid arthritis or osteoarthritis. This report presents a secondary outcome from the primary case-control study examining the relationship of periodontitis with established rheumatoid arthritis. The Centers for Disease Control/American Academy of Periodontology case definitions of periodontitis were used for this analysis. Participants received a full-mouth clinical periodontal examination and panoramic radiograph. Percentage ABHL was measured on posterior teeth. The TLR4 Asp299Gly and CD14/-260 SNPs were selected a priori and genotypes were determined using the ImmunoChip array (Illumina(®) ). Minor allele frequencies and associations with periodontitis and ABHL did not differ according to rheumatoid arthritis vs. osteoarthritis status; therefore, data from these two groups were pooled. The presence of P. gingivalis was detected in subgingival plaque by PCR. Multivariate ordinal

  20. Molecular and clinical analyses of Greig cephalopolysyndactyly and Pallister-Hall syndromes: Robust phenotype prediction from the type and position of GLI3 mutations

    Johnston, Jennifer J.; Olivos-Glander, Isabelle; Killoran, Christina; Elson, Emma; Turner, Joyce T.; Peters, Kathryn F.; Abbott, Margaret H.; Aughton, David J.; Aylsworth, Arthur S.; Bamshad, Michael J.; Booth, Carol; Curry, Cynthia J.; David, Albert; Dinulos, Mary Beth; Flannery, David B.; Fox, Michelle A.; Graham, John M.; Grange, Dorothy K.; Guttmacher, Alan E.; Hannibal, Mark C.; Henn, Wolfram; Hennekam, Raoul C. M.; Holmes, Lewis B.; Hoyme, H. Eugene; Leppig, Kathleen A.; Lin, Angela E.; Macleod, Patrick; Manchester, David K.; Marcelis, Carlo; Mazzanti, Laura; McCann, Emma; McDonald, Marie T.; Mendelsohn, Nancy J.; Moeschler, John B.; Moghaddam, Billur; Neri, Giovanni; Newbury-Ecob, Ruth; Pagon, Roberta A.; Phillips, John A.; Sadler, Laurie S.; Stoler, Joan M.; Tilstra, David; Walsh Vockley, Catherine M.; Zackai, Elaine H.; Zadeh, Touran M.; Brueton, Louise; Black, Graeme Charles M.; Biesecker, Leslie G.

    2005-01-01

    Mutations in the GLI3 zinc-finger transcription factor gene cause Greig cephalopolysyndactyly syndrome (GCPS) and Pallister-Hall syndrome (PHS), which are variable but distinct clinical entities. We hypothesized that GLI3 mutations that predict a truncated functional repressor protein cause PHS and

  1. The Effect of the Arg389Gly Beta-1 Adrenoceptor Polymorphism on Plasma Renin Activity and Heart Rate and the Genotype-Dependent Response to Metoprolol Treatment

    Petersen, Morten; Andersen, Jon T; Jimenez-Solem, Espen

    2012-01-01

    A gene-drug interaction has been indicated between beta-1 selective beta-blockers and the Arg389Gly polymorphism (rs1801253) in the adrenergic beta-1 receptor gene (ADRB1). We studied the effect of the ADRB1 Arg389Gly polymorphism on plasma renin activity (PRA) and heart rate (HR) and the genotype...

  2. A Patient With a Mild Holoprosencephaly Spectrum Phenotype and Heterotaxy and a 1.3 Mb Deletion Encompassing GLI2

    Kevelam, S.H.G.; van Harssel, J.J.; van der Zwaag, B.; Smeets, H.J.; Paulussen, A.D.; Lichtenbelt, K.D.

    2012-01-01

    Loss-of-function mutations of GLI2 are associated with features at the mild end of the holoprosencephaly spectrum, including abnormal pituitary gland formation and/or function, and craniofacial abnormalities. In addition patients may have branchial arch anomalies and polydactyly. Large,

  3. Interaction between the Gly460Trp alpha-adducin gene variant and diuretics on the risk of myocardial infarction

    van Wieren-de Wijer, Diane B M A; Maitland-van der Zee, Anke-Hilse; de Boer, Anthonius; Kroon, Abraham A; de Leeuw, Peter W; Schiffers, Paul; Janssen, Rob G J H; Psaty, Bruce M; van Duijn, Cornelia M; Stricker, Bruno H Ch; Klungel, Olaf H

    INTRODUCTION: The Gly460Trp variant of the alpha-adducin gene has been associated with the salt-sensitive and diuretic responsive form of hypertension. OBJECTIVE: The aim of the study was to determine whether the alpha-adducin 460Trp variant allele modifies the risk-lowering effect of diuretics on

  4. Affinity capillary electrophoresis and quantum mechanical calculations applied to investigation of [Gly(6)]-antamanide binding with sodium and potassium ions

    Pangavhane, Sachin; Böhm, S.; Makrlík, E.; Ruzza, P.; Kašička, Václav

    2017-01-01

    Roč. 38, č. 12 (2017), s. 1551-1559 ISSN 0173-0835 R&D Projects: GA ČR(CZ) GA15-01948S Institutional support: RVO:61388963 Keywords : affinity capillary electrophoresis * density functional theory * [Gly(6)]-antamanide * peptide complex * stability constant Subject RIV: CB - Analytical Chemistry, Separation OBOR OECD: Analytical chemistry Impact factor: 2.744, year: 2016

  5. Inhibition of p70S6K2 down-regulates Hedgehog/GLI pathway in non-small cell lung cancer cell lines

    Kotani Hidehito

    2009-07-01

    Full Text Available Abstract Background The Hedgehog (HH pathway promotes tumorigenesis in a diversity of cancers. Activation of the HH signaling pathway is caused by overexpression of HH ligands or mutations in the components of the HH/GLI1 cascade, which lead to increased transactivation of GLI transcription factors. Although negative kinase regulators that antagonize the activity of GLI transcription factors have been reported, including GSK3β, PKA and CK1s, little is known regarding positive kinase regulators that are suitable for use on cancer therapeutic targets. The present study attempted to identify kinases whose silencing inhibits HH/GLI signalling in non-small cell lung cancer (NSCLC. Results To find positive kinase regulators in the HH pathway, kinome-wide siRNA screening was performed in a NSCLC cell line, A549, harboring the GLI regulatory reporter gene. This showed that p70S6K2-silencing remarkably reduced GLI reporter gene activity. The decrease in the activity of the HH pathway caused by p70S6K2-inhibition was accompanied by significant reduction in cell viability. We next investigated the mechanism for p70S6K2-mediated inhibition of GLI1 transcription by hypothesizing that GSK3β, a negative regulator of the HH pathway, is activated upon p70S6K2-silencing. We found that phosphorylated-GSK3β (Ser9 was reduced by p70S6K2-silencing, causing a decreased level of GLI1 protein. Finally, to further confirm the involvement of p70S6K2 in GLI1 signaling, down-regulation in GLI-mediated transcription by PI3KCA-inhibition was confirmed, establishing the pivotal role of the PI3K/p70S6K2 pathway in GLI1 cascade regulation. Conclusion We report herein that inhibition of p70S6K2, known as a downstream effector of the PI3K pathway, remarkably decreases GLI-mediated transactivation in NSCLC by reducing phosphorylated-GSK3β followed by GLI1 degradation. These results infer that p70S6K2 is a potential therapeutic target for NSCLC with hyperactivated HH/GLI pathway.

  6. Downregulation of the Sonic Hedgehog/Gli pathway transcriptional target Neogenin-1 is associated with basal cell carcinoma aggressiveness.

    Casas, Bárbara S; Adolphe, Christelle; Lois, Pablo; Navarrete, Nelson; Solís, Natalia; Bustamante, Eva; Gac, Patricio; Cabané, Patricio; Gallegos, Ivan; Wainwright, Brandon J; Palma, Verónica

    2017-10-13

    Basal Cell Carcinoma (BCC) is one of the most diagnosed cancers worldwide. It develops due to an unrestrained Sonic Hedgehog (SHH) signaling activity in basal cells of the skin. Certain subtypes of BCC are more aggressive than others, although the molecular basis of this phenomenon remains unknown. We have previously reported that Neogenin-1 (NEO1) is a downstream target gene of the SHH/GLI pathway in neural tissue. Given that SHH participates in epidermal homeostasis, here we analyzed the epidermal expression of NEO1 in order to identify whether it plays a role in adult epidermis or BCC. We describe the mRNA and protein expression profile of NEO1 and its ligands (Netrin-1 and RGMA) in human and mouse control epidermis and in a broad range of human BCCs. We identify in human BCC a significant positive correlation in the levels of NEO1 receptor, NTN-1 and RGMA ligands with respect to GLI1 , the main target gene of the canonical SHH pathway. Moreover, we show via cyclopamine inhibition of the SHH/GLI pathway of ex vivo cultures that NEO1 likely functions as a downstream target of SHH/GLI signaling in the skin. We also show how Neo1 expression decreases throughout BCC progression in the K14-Cre:Ptch1 lox/lox mouse model and that aggressive subtypes of human BCC exhibit lower levels of NEO1 than non-aggressive BCC samples. Taken together, these data suggest that NEO1 is a SHH/GLI target in epidermis. We propose that NEO1 may be important in tumor onset and is then down-regulated in advanced BCC or aggressive subtypes.

  7. Characterization of serine hydroxymethyltransferase GlyA as a potential source of D-alanine in Chlamydia pneumoniae.

    De Benedetti, Stefania; Bühl, Henrike; Gaballah, Ahmed; Klöckner, Anna; Otten, Christian; Schneider, Tanja; Sahl, Hans-Georg; Henrichfreise, Beate

    2014-01-01

    For intracellular Chlamydiaceae, there is no need to withstand osmotic challenges, and a functional cell wall has not been detected in these pathogens so far. Nevertheless, penicillin inhibits cell division in Chlamydiaceae resulting in enlarged aberrant bodies, a phenomenon known as chlamydial anomaly. D-alanine is a unique and essential component in the biosynthesis of bacterial cell walls. In free-living bacteria like Escherichia coli, penicillin-binding proteins such as monofunctional transpeptidases PBP2 and PBP3, the putative targets of penicillin in Chlamydiaceae, cross-link adjacent peptidoglycan strands via meso-diaminopimelic acid and D-Ala-D-Ala moieties of pentapeptide side chains. In the absence of genes coding for alanine racemase Alr and DadX homologs, the source of D-Ala and thus the presence of substrates for PBP2 and PBP3 activity in Chlamydiaceae has puzzled researchers for years. Interestingly, Chlamydiaceae genomes encode GlyA, a serine hydroxymethyltransferase that has been shown to exhibit slow racemization of D- and L-alanine as a side reaction in E. coli. We show that GlyA from Chlamydia pneumoniae can serve as a source of D-Ala. GlyA partially reversed the D-Ala auxotrophic phenotype of an E. coli racemase double mutant. Moreover, purified chlamydial GlyA had racemase activity on L-Ala in vitro and was inhibited by D-cycloserine, identifying GlyA, besides D-Ala ligase MurC/Ddl, as an additional target of this competitive inhibitor in Chlamydiaceae. Proof of D-Ala biosynthesis in Chlamydiaceae helps to clarify the structure of cell wall precursor lipid II and the role of chlamydial penicillin-binding proteins in the development of non-dividing aberrant chlamydial bodies and persistence in the presence of penicillin.

  8. Characterization of serine hydroxymethyltransferase GlyA as a potential source of D-alanine in Chlamydia pneumoniae

    Stefania eDe Benedetti

    2014-02-01

    Full Text Available For intracellular Chlamydiaceae, there is no need to withstand osmotic challenges, and a functional cell wall has not been detected in these pathogens so far. Nevertheless, penicillin inhibits cell division in Chlamydiaceae resulting in enlarged aberrant bodies, a phenomenon known as chlamydial anomaly.D-alanine is a unique and essential component in the biosynthesis of bacterial cell walls. In free-living bacteria like Escherichia coli, penicillin-binding proteins such as monofunctional transpeptidases PBP2 and PBP3, the putative targets of penicillin in Chlamydiaceae, cross-link adjacent peptidoglycan strands via meso-diaminopimelic acid and D-Ala-D-Ala moieties of pentapeptide side chains. In the absence of genes coding for alanine racemase Alr and DadX homologs, the source of D-Ala and thus the presence of substrates for PBP2 and PBP3 activity in Chlamydiaceae has puzzled researchers for years. Interestingly, Chlamydiaceae genomes encode GlyA, a serine hydroxymethyltransferase that has been shown to exhibit slow racemization of D- and L- alanine as a side reaction in E. coli. We show that GlyA from Chlamydia pneumoniae can serve as a source of D-Ala. GlyA partially reversed the D-Ala auxotrophic phenotype of an E. coli racemase double mutant. Moreover, purified chlamydial GlyA had racemase activity on L-Ala in vitro and was inhibited by D-cycloserine, identifying GlyA, besides D-Ala ligase MurC/Ddl, as an additional target of this competitive inhibitor in Chlamydiaceae. Proof of D-Ala biosynthesis in Chlamydiaceae helps to clarify the structure of cell wall precursor lipid II and the role of chlamydial penicillin-binding proteins in the development of non-dividing aberrant chlamydial bodies and persistence in the presence of penicillin.

  9. Reciprocal Regulation of Hypoxia-Inducible Factor 2α and GLI1 Expression Associated With the Radioresistance of Renal Cell Carcinoma

    Zhou, Jiancheng [Department of Urology, First Affiliated Hospital of Medical School, Xi' an Jiaotong University, Xi' an (China); Department of Urology, Department of Radiation Oncology, University of Texas Southwestern Medical Center, Dallas, Texas (United States); Wu, Kaijie [Department of Urology, First Affiliated Hospital of Medical School, Xi' an Jiaotong University, Xi' an (China); Gao, Dexuan [Department of Urology, Shandong Provincial Hospital affiliated with Shandong University, Ji' nan (China); Zhu, Guodong; Wu, Dapeng; Wang, Xinyang; Chen, Yule; Du, Yuefeng; Song, Wenbin; Ma, Zhenkun [Department of Urology, First Affiliated Hospital of Medical School, Xi' an Jiaotong University, Xi' an (China); Authement, Craig; Saha, Debabrata [Department of Urology, Department of Radiation Oncology, University of Texas Southwestern Medical Center, Dallas, Texas (United States); Hsieh, Jer-Tsong, E-mail: jt.hsieh@utsouthwestern.edu [Department of Urology, Department of Radiation Oncology, University of Texas Southwestern Medical Center, Dallas, Texas (United States); He, Dalin, E-mail: dalinhe@yahoo.com [Department of Urology, First Affiliated Hospital of Medical School, Xi' an Jiaotong University, Xi' an (China)

    2014-11-15

    Purpose: Renal cell carcinoma (RCC) is often considered a radioresistant tumor, but the molecular mechanism underlying its radioresistance is poorly understood. This study explored the roles of hypoxia-inducible factor 2α (HIF2α) and sonic hedgehog (SHH)-GLI1 signaling in mediating the radioresistance of RCC cells and to unveil the interaction between these 2 signaling pathways. Methods and Materials: The activities of SHH-GLI1 signaling pathway under normoxia and hypoxia in RCC cells were examined by real-time polymerase chain reaction, Western blot, and luciferase reporter assay. The expression of HIF2α and GLI1 in RCC patients was examined by immunohistochemistry, and their correlation was analyzed. Furthermore, RCC cells were treated with HIF2α-specific shRNA (sh-HIF2α), GLI1 inhibitor GANT61, or a combination to determine the effect of ionizing radiation (IR) on RCC cells based on clonogenic assay and double-strand break repair assay. Results: RCC cells exhibited elevated SHH-GLI1 activities under hypoxia, which was mediated by HIF2α. Hypoxia induced GLI1 activation through SMO-independent pathways that could be ablated by PI3K inhibitor or MEK inhibitor. Remarkably, the SHH-GLI1 pathway also upregulated HIF2α expression in normoxia. Apparently, there was a positive correlation between HIF2α and GLI1 expression in RCC patients. The combination of sh-HIF2α and GLI1 inhibitor significantly sensitized RCC cells to IR. Conclusions: Cross-talk between the HIF2α and SHH-GLI1 pathways was demonstrated in RCC. Cotargeting these 2 pathways, significantly sensitizing RCC cells to IR, provides a novel strategy for RCC treatment.

  10. Investigation of possible association between Ser9Gly polymorphism of the D3 dopaminergic receptor gene and response to typical antipsychotics in patients with schizophrenia

    Quirino Cordeiro

    Full Text Available Typical antipsychotics have a high affinity for dopamine receptors. It is therefore of interest to investigate such loci in pharmacogenetic studies on psychosis. We investigated the hypothesis that Ser9Gly polymorphism of the DRD3 gene may play a role in the differences in individual response to typical antipsychotics between schizophrenic patients. The sample was composed of 53 good responders and 59 poor ones. No significant differences between the good and poor responders were found in the allelic distribution (good responders: Ser9 61.32%, Gly9 38.67%; poor responders: Ser9 64.40%, Gly9 35.59%; odds ratio, OR = 0.88, 0.49 < OR < 1.56; chi2 = 0.23, 1 degree of freedom, df, p = 0.63 and genotype distribution (good responders: Ser9/Ser9 37.73%, Ser9/Gly9 47.16%, Gly9/Gly9 15.09%; poor responders: Ser9/Ser9 42.37%, Ser9/Gly9 44.06%, Gly9/Gly9 13.55%; chi2 = 0.25, 2 df, p = 0.88. Nor was there any association with homozygosity (good responders: homozygous: 52.82%, heterozygous: 47.16%; poor responders: homozygous: 55.92%, heterozygous: 44.06%; odds ratio, OR = 0.88, 0.39 < OR < 1.99; chi2 = 0.11, 1 df, p = 0.74. The results did not support the hypothesis that Ser9Gly polymorphism of the DRD3 gene influences the response to typical antipsychotics in our sample of schizophrenics.

  11. Attraverso gli studi spagnoli di Cesare de Lollis (1887-1924

    Diego Stefanelli

    2016-07-01

    Full Text Available L’articolo considera gli studi di ispanistica di Cesare De Lollis nel loro complesso: dalla fine dell’Ottocento, con gli articoli sulla lirica ispano-portoghese medievale (di stampo filologico-erudito e gli interventi (di tipo giornalistico-divulgativo su alcuni scrittori spagnoli dell’Ottocento, agli anni Venti del Novecento, con il volume su Cervantes reazionario (1924. Un momento importante di questo percorso fu il passaggio di De Lollis alla cattedra romana di «Letterature francese e spagnola moderne» (avvenuto nel 1905: ricostruite le vicende della cattedra, si pone l’attenzione sugli scritti coevi di De Lollis incentrati sulla necessità di uno studio serio delle letterature moderne. Vengono poi studiate le note e le recensioni di letteratura spagnola apparse nelle prime decadi del secolo, in particolare lo scritto Classicismo e secentismo (1908 su Fernando de Herrera, esponente di quella poesia «eroica» studiata a piú riprese da De Lollis (in particolare per il contesto francese. L’articolo si concentra poi sul volume Cervantes reazionario, mostrando che molte delle questioni trattate erano presenti già nel primo articolo del 1913, contemporaneo a quelli sui romantici italiani confluiti poi nei Saggi sulla forma poetica italiana dell’Ottocento. Proprio il nesso tra questi scritti e quelli su Cervantes consente di leggere il volume cervantino alla luce di un tema centrale della critica di De Lollis: la contrapposizione tra l’«eroe» della poesia classicistica e il «realismo» della rivoluzione romantica.  The paper deals with Cesare De Lollis’ Hispanic Studies from the end of the 19th Century (with the philological studies on Spanish-Portuguese poetry of the Middle Age and the coeval journalistic articles on Spanish writers of the 19th Century to the book Cervantes reazionario (1924. A turning moment in De Lollis’ approach to Spanish literature is 1905, when he obtained the chair of «Modern French and Spanish

  12. The Hedgehog-GLI pathway in embryonic development and cancer: implications for pulmonary oncology therapy

    Armas-López, Leonel; Zúñiga, Joaquín; Arrieta, Oscar; Ávila-Moreno, Federico

    2017-01-01

    Transcriptional regulation and epigenetic mechanisms closely control gene expression through diverse physiological and pathophysiological processes. These include the development of germ layers and post-natal epithelial cell-tissue differentiation, as well as, involved with the induction, promotion and/or progression of human malignancies. Diverse studies have shed light on the molecular similarities and differences involved in the stages of embryological epithelial development and dedifferentiation processes in malignant tumors of epithelial origin, of which many focus on lung carcinomas. In lung cancer, several transcriptional, epigenetic and genetic aberrations have been described to partly arise from environmental risk factors, but ethnic genetic predisposition factors may also play a role. The classification of the molecular hallmarks of cancer has been essential to study and achieve a comprehensive view of the interaction networks between cell signaling pathways and functional roles of the transcriptional and epigenetic regulatory mechanisms. This has in turn increased understanding on how these molecular networks are involved in embryo-layers and malignant diseases development. Ultimately, a major biomedicine goal is to achieve a thorough understanding of their roles as diagnostic, prognostic and treatment response indicators in lung oncological patients. Recently, several notable cell-signaling pathways have been studied based on their contribution to promoting and/or regulating the engagement of different cancer hallmarks, among them genome instability, exacerbated proliferative signaling, replicative immortality, tumor invasion-metastasis, inflammation, and immune-surveillance evasion mechanisms. Of these, the Hedgehog-GLI (Hh) cell-signaling pathway has been identified as a main molecular contribution into several of the abovementioned functional embryo-malignancy processes. Nonetheless, the systematic study of the regulatory epigenetic and

  13. New zinc-glycine-iodide complexes as a product of equilibrium and non-equilibrium crystallization in the Gly – ZnI2 – H2O system

    Tepavitcharova, S.; Havlíček, D.; Matulková, I.; Rabadjieva, D.; Gergulova, R.; Plocek, Jiří; Němec, I.; Císařová, I.

    2016-01-01

    Roč. 1120, SEP (2016), s. 42-49 ISSN 0022-2860 Institutional support: RVO:61388980 Keywords : [Zn(gly)I2], [Zn(gly)2I2] * [Zn3(H2O)4(μ-gly)2I6] * Crystal structure * Vibrational spectra * Thermal behaviour Subject RIV: CA - Inorganic Chemistry Impact factor: 1.753, year: 2016

  14. Anti-arrhythmic peptide N-3-(4-hydroxyphenyl)propionyl Pro-Hyp-Gly-Ala-Gly-OH reduces dispersion of action potential duration during ischemia/reperfusion in rabbit hearts

    Kjølbye, Anne Louise; Petersen, Jørgen Søberg; Holstein-Rathlou, N.-H.

    2002-01-01

    During ischemia, cardiac gap junctions close and neighboring cells uncouple. This leads to slow conduction, increased dispersion of APD90 (duration from action potential beginning to 90% of repolarization), nonuniform anisotropy, and unidirectional conduction block, all of which favor the induction...... of reentry arrhythmias. It has been suggested that anti-arrhythmic peptides increase gap junction conductance during states of reduced coupling. The aim of this study was to test the effect of the anti-arrhythmic peptide N-3-(4-hydroxyphenyl)propionyl Pro-Hyp-Gly-Ala-Gly-OH (HP-5) (10(-10) ) on dispersion...... of epicardial APD90 during both normokalemic and hypokalemic ischemia/reperfusion in isolated perfused rabbit hearts. HP-5 did not affect average APD90, heart rate, left ventricular contractility (LVP dP/dtmax), or mean coronary flow. HP-5 significantly reduced the epicardial APD dispersion during hypokalemic...

  15. L’arcipelago postmoderno. Oreste del Buono e gli anni Settanta

    Andrea Chiurato

    2012-12-01

    Il tentativo di cooptazione postuma tra le file dei simpatizzanti da parte del Gruppo 63 avrà breve fortuna (verrà incluso nella prima antologia del gruppo, ma non nelle se-guenti, e Renato Barilli arriverà addirittura a porlo a capo di un improbabile «Polo Bu-tor» (Chiurato 14, uno sparuto gruppo di „infiltrati‟ (insieme a Raffaele La Capria, Emilio Tadini, e Giuliano Gramigna nelle file della compagine neoavanguardista. Quarant‟anni dopo la questione rimane aperta e l‟eredità di quel turbolento periodo è ancora oggetto di discussione. La nostra intenzione è proporre un approccio diverso, che cer-chi di restituire la fluidità e la ricchezza delle suggestioni di allora poi irrigiditesi in cate-gorie critiche spesso frutto di semplificazioni indebite. Del Buono, irriducibile outsider, ci permette così di riscoprire gli anni Settanta da un‟angolazione inedita e di colmare, nello stesso tempo, alcune improvvide lacune nella nostra memoria culturale

  16. Il degrado abusivo e il "degrado legale" del territorio: gli ecomostri

    Anna Cicchetti

    2010-07-01

    Full Text Available Il fenomeno di una urbanizzazione senza criteri né misure, collegata a ragioni turistiche, molto spesso opportunistiche, è affrontato dalla giurisprudenza nella sua azione di controllo del territorio: ne sono un caso esemplare gli “ecomostri”, ormai comunemente e tristemente sono conosciuti dalla società civile. Queste costruzioni di grande impatto territoriale, ambientale e paesaggistico, in alcuni casi sono legittimati da interventi normativi non bene soppesati e in altri avvengono in maniera c.d. abusiva, con l’unica conseguenza, in entrambe le ipotesi, di deturpare l’ambiente circostante sotto molteplici profili, non ultimo quello incidente su località turistiche. Il contributo si pone l’obiettivo di contribuire ad una politica turistica attenta al profilo ambientale: occorre individuare una maggiore cogenza e conoscenza normativa a favore di una pianificazione urbanistica e politica del paesaggio, in grado di sostenere il recupero, la riqualificazione e valorizzazione delle bellezze naturali presenti sul territorio. Solo il coinvolgimento e la necessaria collaborazione di tutti i soggetti del territorio per una programmazione orientata a realizzare uno «sviluppo capace di rispondere ai bisogni delle generazioni attuali, senza compromettere la capacità delle generazioni future di rispondere ai propri».

  17. La misurazione dei processi di regolazione cognitiva e metacognitiva durante lo studio con gli ipermedia

    Roger Azevedo

    2010-01-01

    Full Text Available L’apprendimento autoregolato rappresenta una modalità di apprendimento di fondamentale importanza quando ci si avvale del supporto di ambienti ipermediali. Obiettivo di questo articolo è presentare quattro assunzioni chiave che consentono la misurazione dei processi cognitivi e metacognitivi attivati durante l’apprendimento tramite ipermedia. Innanzi tutto, assumiamo che sia possibile individuare, tracciare, modellare e favorire processi di apprendimento auto-regolato durante lo studio con gli ipermedia. La seconda assunzione si focalizza sul comprendere come la complessità dei processi regolatori che avvengono durante l’apprendimento mediato da sistemi ipermediali sia importante per determinare il perché alcuni processi vengono messi in atto durante l’esecuzione di un compito. Le terza assunzione è relativa al considerare che l’utilizzo di processi di apprendimento auto-regolato possa dinamicamente cambiare nel tempo e che tali processi sono di natura ciclica (influenzati dalle condizioni interne ed esterne e da meccanismi di feedback. Infine, l’identificazione, raccolta e classificazione dei processi di apprendimento autoregolato utilizzati durante lo studio con sistemi ipermediali, può risultare un compito alquanto difficoltoso.

  18. Manganese induces mitochondrial dynamics impairment and apoptotic cell death: a study in human Gli36 cells.

    Alaimo, Agustina; Gorojod, Roxana M; Miglietta, Esteban A; Villarreal, Alejandro; Ramos, Alberto J; Kotler, Mónica L

    2013-10-25

    Manganese (Mn) is an essential trace element due to its participation in many physiological processes. However, overexposure to this metal leads to a neurological disorder known as Manganism whose clinical manifestations and molecular mechanisms resemble Parkinson's disease. Several lines of evidence implicate astrocytes as an early target of Mn neurotoxicity being the mitochondria the most affected organelles. The aim of this study was to investigate the possible mitochondrial dynamics alterations in Mn-exposed human astrocytes. Therefore, we employed Gli36 cells which express the astrocytic markers GFAP and S100B. We demonstrated that Mn triggers the mitochondrial apoptotic pathway revealed by increased Bax/Bcl-2 ratio, by the loss of mitochondrial membrane potential and by caspase-9 activation. This apoptotic program may be in turn responsible of caspase-3/7 activation, PARP-1 cleavage, chromatin condensation and fragmentation. In addition, we determined that Mn induces deregulation in mitochondria-shaping proteins (Opa-1, Mfn-2 and Drp-1) expression levels in parallel with the disruption of the mitochondrial network toward to an exacerbated fragmentation. Since mitochondrial dynamics is altered in several neurodegenerative diseases, these proteins could become future targets to be considered in Manganism treatment. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  19. Interaction of the univalent silver cation with [Gly6]-antamanide: Experimental and theoretical study

    Makrlík, Emanuel; Böhm, Stanislav; Kvíčala, Jaroslav; Vaňura, Petr; Ruzza, Paolo

    2018-03-01

    On the basis of extraction experiments and γ-activity measurements, the extraction constant corresponding to the equilibrium Ag+(aq) + 1.Na+(nb) ⇄ 1.Ag+ (nb) + Na+(aq) occurring in the two-phase water - nitrobenzene system (1 = [Gly6]-antamanide; aq = aqueous phase, nb = nitrobenzene phase) was determined as log Kex (Ag+,1·Na+) = 1.5 ± 0.1. Further, the stability constant of the 1·Ag+ complex in nitrobenzene saturated with water was calculated for a temperature of 25 °C: log βnb (1·Ag+) = 4.5 ± 0.2. Finally, by using quantum chemical DFT calculations, the most probable structure of the cationic complex species 1·Ag+ was derived. In the resulting complex, the "central" cation Ag+ is coordinated by four noncovalent interactions to the corresponding four carbonyl oxygen atoms of the parent ligand 1. Besides, the whole 1·Ag+ complex structure is stabilized by two intramolecular hydrogen bonds. The interaction energy of the considered 1·Ag+ complex was found to be -465.5 kJ/mol, confirming also the formation of this cationic species.

  20. Arg16Gly and Gln27Glu β2 adrenergic polymorphisms influence cardiac autonomic modulation and baroreflex sensitivity in healthy young Brazilians

    Atala, Magda M; Goulart, Alessandra; Guerra, Grazia M; Mostarda, Cristiano; Rodrigues, Bruno; Mello, Priscila R; Casarine, Dulce E; Irigoyen, Maria-Claudia; Pereira, Alexandre C; Consolim-Colombo, Fernanda M

    2015-01-01

    The association between functional β2 adrenergic receptor (β2-AR) polymorphisms and cardiac autonomic modulation is still unclear. Thus, two common polymorphisms in the β2-AR gene (Gln27Glu β2 and Arg16Gly β2) were studied to determine whether they might affect tonic and reflex cardiac sympathetic activity in healthy young subjects. A total of 213 healthy young white subjects of both genders (53% female), aged 18-30 years (23.5±3.4 y), had their continuous blood pressure curves noninvasively recorded by Finometer at baseline, and other hemodynamic parameters, as cardiac autonomic modulation, baroreflex sensitivity, and allele, genotype, and diplotype frequencies calculated. Associations were made between Arg16Gly β2 and Gln27Glu β2 polymorphisms and between β2-AR diplotypes and all variables. The heart rate was significantly lower (P<0.001) in the presence of homozygous Arg/Arg alleles (60.9±1.5 bpm) than in that of Arg/Gly heterozygotes (65.9±1.0 bpm) or Gly/Gly homozygotes (66.3±1.2 bpm). Homozygous carriers of Arg16 allele had an alpha index (19.2±1.3) significantly higher (P<0.001) than that of the subjects with the Gly allele Gly/Gly (14.5±0.7) or Arg/Gly (14.6±0.7). Furthermore, the recessive Glu27Glu and the heterozygous Gln27Glu genotypes had a higher percentage of low-frequency components (LF%) than the homozygous Gln27Gln (15.1% vs. 16.0% vs. 8.2%, P=0.03, respectively). In healthy young subjects, the presence of β2-AR Arg16 allele in a recessive model was associated with higher baroreflex sensitivity, and increased parasympathetic modulation in studied individuals. PMID:25755837

  1. Meanings of a Territorial Infrastructure: the Airports Significati di una infrastruttura territoriale: gli aeroporti

    Giuseppe Mazzeo

    2011-11-01

    lacking of physical and spatial integration with their urban context; for this the airports have evolved into semi-autonomous spaces. For their dimensional characteristics and for the incidence of pollutants factors, must be placed outside the city, often at significant distances from the same: their fundamental role in the urban structure is expressed, therefore, in external spaces from the city. As defined above, the airport is an urban infrastructure. But it is other things also: it is an ordered place, because its management requires defined actions and sequences of activities that are the true opposite of the urban disorder. Both the structures are complexes but the airport is a side where the hierarchical roles are well defined, more than in other sides, and it is the side where the authority is recognized and well accepted. The building of new airports, or the modernization of existing structures are a sign of the sectors’ vitality and of the necessity to offer better services to an expanding custom. But the airports have created a lot of imagines around their presence and use.

    mplessi, ma il primo è uno spazio in cui i ruoli gerarchici sono ben definiti e dove è l'autorità è riconosciuta e ben accettata.

     

    La costruzione di nuovi aeroporti, o l'ammodernamento delle strutture esistenti, è un segno della vitalità del settore e della necessità di offrire migliori servizi ad una utenza in espansione.

    L’articolo vuole esplorare i significati principali di una infrastruttura come l’aeroporto. Essa è una infrastruttura territoriale che aumenta l'accessibilità spaziale e il potenziale economico di un territorio; in questo senso, è una porta per incrementare gli scambi turistici e culturali. È anche una infrastruttura che porta con sé gli investimenti in altre aree della mobilità. A partire dagli anni Sessanta, inoltre, è diventato una icona e un luogo simbolico dove sperimentare nuove soluzioni architettoniche. Sono fattori limitativi

  2. The Brachyury Gly177Asp SNP Is not Associated with a Risk of Skull Base Chordoma in the Chinese Population

    Zhen Wu

    2013-10-01

    Full Text Available A recent chordoma cancer genotyping study reveals that the rs2305089, a single nucleotide polymorphism (SNP located in brachyury gene and a key gene in the development of notochord, is significantly associated with chordoma risk. The brachyury gene is believed to be one of the key genes involved in the pathogenesis of chordoma, a rare primary bone tumor originating along the spinal column or at the base of the skull. The association between the brachyury Gly177Asp single nucleotide polymorphism (SNP and the risk of skull base chordoma in Chinese populations is currently unknown. We investigated the genotype distribution of this SNP in 65 skull-base chordoma cases and 120 healthy subjects. Comparisons of the genotype distributions and allele frequencies did not reveal any significant difference between the groups. Our data suggest that the brachyury Gly177Asp SNP is not involved in the risks of skull-base chordoma, at least in the Chinese population.

  3. Cyclization of the N-Terminal X-Asn-Gly Motif during Sample Preparation for Bottom-Up Proteomics

    Zhang, Xumin; Højrup, Peter

    2010-01-01

    We, herein, report a novel -17 Da peptide modification corresponding to an N-terminal cyclization of peptides possessing the N-terminal motif of X-Asn-Gly. The cyclization occurs spontaneously during sample preparation for bottom-up proteomics studies. Distinct from the two well-known N-terminal ......We, herein, report a novel -17 Da peptide modification corresponding to an N-terminal cyclization of peptides possessing the N-terminal motif of X-Asn-Gly. The cyclization occurs spontaneously during sample preparation for bottom-up proteomics studies. Distinct from the two well-known N......-terminal cyclizations, cyclization of N-terminal glutamine and S-carbamoylmethylcysteine, it is dependent on pH instead of [NH(4)(+)]. The data set from our recent study on large-scale N(α)-modified peptides revealed a sequence requirement for the cyclization event similar to the well-known deamidation of Asn to iso...

  4. Affinity capillary electrophoresis and density functional theory study of noncovalent interactions of cyclic peptide [Gly(6)]-antamanide with small cations

    Pangavhane, Sachin; Böhm, S.; Makrlík, E.; Ruzza, P.; Kašička, Václav

    2017-01-01

    Roč. 38, č. 16 (2017), s. 2025-2033 ISSN 0173-0835 R&D Projects: GA ČR(CZ) GA15-01948S Institutional support: RVO:61388963 Keywords : affinity capillary electrophoresis * cesium complex * density functional theory * [Gly(6)]-antamanide * rubidium complex * stability constants Subject RIV: CB - Analytical Chemistry, Separation OBOR OECD: Analytical chemistry Impact factor: 2.744, year: 2016

  5. Sall4-Gli3 system in early limb progenitors is essential for the development of limb skeletal elements

    Akiyama, Ryutaro; Kawakami, Hiroko; Wong, Julia; Oishi, Isao; Nishinakamura, Ryuichi; Kawakami, Yasuhiko

    2015-01-01

    The limb skeletal elements that have unique morphology and distinct locations are developed from limb progenitors, derived from the lateral plate mesoderm. These skeletal elements arise during limb development. In this study, we show genetic evidence that function of Sall4 is essential prior to limb outgrowth for development of the anterior-proximal skeletal elements. Furthermore, genetic interaction between Sall4 and Gli3 is upstream of establishing Shh (Sonic hedgehog) expression, and there...

  6. Structure-activity relationship of linear peptide Bu-His-DPhe-Arg-Trp-Gly-NH(2) at the human melanocortin-1 and -4 receptors: histidine substitution.

    Cheung, Adrian Wai-Hing; Danho, Waleed; Swistok, Joseph; Qi, Lida; Kurylko, Grazyna; Rowan, Karen; Yeon, Mitch; Franco, Lucia; Chu, Xin-Jie; Chen, Li; Yagaloff, Keith

    2003-01-06

    Systematic substitution of His(6) residue using non-selective hMC4R pentapeptide agonist (Bu-His(6)-DPhe(7)-Arg(8)-Trp(9)-Gly(10)-NH(2)) as the template led to the identification of Bu-Atc(6)(2-aminotetraline-2-carboxylic acid)-DPhe(7)-Arg(8)-Trp(9)-Gly(10)-NH(2) which showed moderate selectivity towards hMC4R over hMC1R. Further SAR studies resulted in the discovery of Penta-5-BrAtc(6)-DPhe(7)-Arg(8)-Trp(9)-Gly(10)-NH(2) and Penta-5-Me(2)NAtc(6)-DPhe(7)-Arg(8)-Trp(9)-Gly(10)-NH(2) which are potent hMC4R agonists and are inactive in hMC1R, hMC3R and hMC5R agonist assays.

  7. Intestinal drug transport via the proton-coupled amino acid transporter PAT1 (SLC36A1) is inhibited by Gly-X(aa) dipeptides

    Frølund, Sidsel; Langthaler, Louise; Kall, Morten A

    2012-01-01

    -Sar as substrates of the amino acid transporter PAT1. The aim of the present study is to investigate if other Gly-containing dipeptides interact with PAT1, and whether they can inhibit PAT1 mediated drug absorption, in vitro and in vivo. The in vitro methods included two-electrode voltage clamp measurements on h...... of different dipeptides. The in vivo part consisted of a pharmacokinetic study in rats following oral administration of gaboxadol and preadministration of 200 mg/kg dipeptide. The results showed that in hPAT1 expressing oocytes Gly-Tyr, Gly-Pro, and Gly-Phe inhibited currents induced by drug substances......, the present study identifies selected dipeptides as inhibitors of PAT1 mediated drug absorption in various in vitro models....

  8. Activity of Metabotropic Glutamate Receptor 4 Suppresses Proliferation and Promotes Apoptosis With Inhibition of Gli-1 in Human Glioblastoma Cells

    Zhichao Zhang

    2018-05-01

    Full Text Available Glioblastoma multiforme (GBM is the most lethal glioma variant in the adult brain and among the deadliest of human cancers. Increasing evidence has shown that metabotropic glutamate receptor subtype 4 (mGluR4 expression may play roles in regulating the growth of neural stem cells as well as several cancer cell lines. Here, we investigated the effects of mGluR4 on the growth and apoptosis of the LN229 GBM cell line. Involvement of Gli-1, one of the key transcription factors in the sonic Hedgehog (SHH signaling pathway, was further explored. In this study, mGluR4 was activated using selective agonist VU0155041; and gene-targeted siRNAs were used to generate loss of function of mGluR4 and Gli-1 in LN229 cells. The results demonstrated that LN229 cells expressed mGluR4 and the agonist VU0155041 decreased cell viability in a dose- and time-dependent manner. Activation of mGluR4 inhibited cyclin D1 expression, activated pro-caspase-8/9/3, and disrupted the balance of Bcl-2/Bax expression, which indicated cell cycle arrest and apoptosis of LN229 cells, respectively. Furthermore, Gli-1 expression was reduced by mGluR4 activation in LN229 cells, and downregulation of Gli-1 expression by gene-targeted siRNA resulted in both inhibition of cell proliferation and promotion of apoptosis. Moreover, VU0155041 treatment substantially blocked SHH-induced cyclin D1 expression and cell proliferation, while increasing TUNEL-positive cells and the activation of apoptosis-related proteins. We concluded that activation of mGluR4 expressed in LN229 cells could inhibit GBM cell growth by decreasing cell proliferation and promoting apoptosis. Further suppression of intracellular Gli-1 expression might be involved in the action of mGluR4 on cancer cells. Our study suggested a novel role of mGluR4, which might serve as a potential drug target for control of GBM cell growth.

  9. Analysis of the Gli-D2 locus identifies a genetic target for simultaneously improving the breadmaking and health-related traits of common wheat.

    Li, Da; Jin, Huaibing; Zhang, Kunpu; Wang, Zhaojun; Wang, Faming; Zhao, Yue; Huo, Naxin; Liu, Xin; Gu, Yong Q; Wang, Daowen; Dong, Lingli

    2018-05-11

    Gliadins are a major component of wheat seed proteins. However, the complex homoeologous Gli-2 loci (Gli-A2, -B2 and -D2) that encode the α-gliadins in commercial wheat are still poorly understood. Here we analyzed the Gli-D2 locus of Xiaoyan 81 (Xy81), a winter wheat cultivar. A total of 421.091 kb of the Gli-D2 sequence was assembled from sequencing multiple bacterial artificial clones, and 10 α-gliadin genes were annotated. Comparative genomic analysis showed that Xy81 carried only eight of the α-gliadin genes of the D genome donor Aegilops tauschii, with two of them each experiencing a tandem duplication. A mutant line lacking Gli-D2 (DLGliD2) consistently exhibited better breadmaking quality and dough functionalities than its progenitor Xy81, but without penalties in other agronomic traits. It also had an elevated lysine content in the grains. Transcriptome analysis verified the lack of Gli-D2 α-gliadin gene expression in DLGliD2. Furthermore, the transcript and protein levels of protein disulfide isomerase were both upregulated in DLGliD2 grains. Consistent with this finding, DLGliD2 had increased disulfide content in the flour. Our work sheds light on the structure and function of Gli-D2 in commercial wheat, and suggests that the removal of Gli-D2 and the gliadins specified by it is likely to be useful for simultaneously enhancing the end-use and health-related traits of common wheat. Because gliadins and homologous proteins are widely present in grass species, the strategy and information reported here may be broadly useful for improving the quality traits of diverse cereal crops. © 2018 The Authors The Plant Journal © 2018 John Wiley & Sons Ltd.

  10. Efficient Absorption of X-Hydroxyproline (Hyp)-Gly after Oral Administration of a Novel Gelatin Hydrolysate Prepared Using Ginger Protease.

    Taga, Yuki; Kusubata, Masashi; Ogawa-Goto, Kiyoko; Hattori, Shunji

    2016-04-13

    Recent studies have reported that oral intake of gelatin hydrolysate has various beneficial effects, such as reduction of joint pain and lowering of blood sugar levels. In this study, we produced a novel gelatin hydrolysate using a cysteine-type ginger protease having unique substrate specificity with preferential peptide cleavage with Pro at the P2 position. Substantial amounts of X-hydroxyproline (Hyp)-Gly-type tripeptides were generated up to 2.5% (w/w) concomitantly with Gly-Pro-Y-type tripeptides (5%; w/w) using ginger powder. The in vivo absorption of the ginger-degraded gelatin hydrolysate was estimated using mice. The plasma levels of collagen-derived oligopeptides, especially X-Hyp-Gly, were significantly high (e.g., 2.3-fold for Glu-Hyp-Gly, p < 0.05) compared with those of the control gelatin hydrolysate, which was prepared using gastrointestinal proteases and did not contain detectable X-Hyp-Gly. This study demonstrated that orally administered X-Hyp-Gly was effectively absorbed into the blood, probably due to the high protease resistance of this type of tripeptide.

  11. Le maschere senza volto. Gli eroi “tragici” del nouveau roman

    Andrea Chiurato

    2017-11-01

    Full Text Available Tra i vari movimenti sperimentali affermatisi nel corso del secondo Novecento il nouveau roman offre un’occasione quanto mai preziosa a chi volesse riconsiderare il mutevole valore e le innumerevoli funzioni attribuite al “tragico” alle soglie dell’era postmoderna. Posta al centro del dibattito critico da Alain Robbe-Grillet, la riflessione sull’ambiguo statuto di questo peculiare genere ha d’altronde rappresentato un momento cruciale nella definizione dell’identità dell’école du regard e del suo peculiare rapporto con la tradizione del canone occidentale. Robbe-Grillet, dal canto suo, pur ispirandosi alla vicenda di Edipo nella sua opera d’esordio, non esiterà in seguito a condannare la tragedia come ingannevole forma di compromesso tra l’uomo e le choses e a rifiutare, insieme a essa, la “metafisica della presenza” di cui sarebbe espressione. Altri nouveaux romanciers – tra cui Michel Butor – preferiranno invece sfruttarne le potenzialità in maniera inedita, ritenendola uno sfondo mitopoietico ideale su cui far emergere i tratti essenziali di una condizione esistenziale all’insegna del crollo delle grandi narrazioni. Al di là delle diverse intenzioni, la rivisitazione della “forma” in sé (in chiave parodica o attualizzante ha di conseguenza svolto un ruolo cruciale sia nella decostruzione del “personaggio-uomo”, sia nella contestazione delle cosiddette “nozioni scadute” tanto indigeste a questa eterogenea collection d’écrivains. Da Les gommes (1953 a L’emploi du temps (1956, per arrivare sino a Sphinx en chapeau (1984, si potrebbero menzionare numerosi esempi a tal riguardo, segno di una fascinazione costante, ma non per questo meno conflittuale, destinata a esercitare una profonda influenza su alcuni dei maggiori dei successi editoriali dello sperimentalismo francese. Un’influenza di cui oggi, ormai esauritisi gli accesi toni dello scandalo e della provocazione che accompagnarono l

  12. Synthesis of Fluorine-18 Labeled Glucose-Lys-Arg-Gly-Asp-D-Phe as a Potential Tumor Imaging Agent

    Lee, Kyo Chul; Kim, Ji Sun; Sung, Hyun Ju; Jung, Jae Ho; An, Gwang Il; Chi, Dae Yoon; Lee, Byung Chul; Moon, Byung Seok; Choi, Tae Hyun; Chuna, Kwon Soo

    2005-01-01

    The α v β 3 integrin is an important receptor affecting tumor growth, metastatic potential on proliferating endothelial cells as well as on tumor cells of various origin, tumor-induced angiogenesis could be blocked by antagonizing the α v β 3 integrin with RGD. Therefore, α v β 3 integrin is a target for angiogenesis imaging that might be useful in assessing tumor-induced angiogenesis and identifying tumor metastasis. To design potent radiotracer for imaging angiogenesis containing a cRGD moiety should include low hepatic uptake in vivo. Tripeptide Arg-Gly-Asp (RGD), naturally existed in extracellular matrix proteins, is known to be the primary binding site of the α v β 3 integrin. The imaging of α v β 3 receptor expression will give the information of the metastatic ability of the tumor which is not available by [ 18 F]FDG. Our interest in developing new radiopharmaceuticals for in vivo visualization of angiogenesis has led us to synthesize derivatives of cRGD (cyclic arginineglycine-aspartic acid) that contains glucose moiety. Because sugar-protein interaction is a key step in metastasis and angiogenesis, it has also been proposed to play an intriguing role in imaging of tumor. We designed and synthesized two fluorine-18 labeled RGD glycopeptides . N-fluorobenzyl-diaminobutane-N'-glucose-Lys-Arg-Gly-Asp-D-Phe ([ 18 F]fluorobenzyl-glucose-KRGDf, and Nfluorobenzoyl- diaminobutane-N'-glucose-Lys-Arg-Gly-Asp-D-Phe ([ 18 F]fluorobenzoyl-glucose-KRGDf, from same precursor as a diagnostic tumor imaging agent for positron emission tomography (PET). Fluorine-18 labeled cRGD glycopeptides were prepared using two different simple labeling methods: one is reductive alkylation of an amine with [ 18 F]fluorobenzaldehyde and the other is amide condensation with [ 18 F]fluorobenzoic acid

  13. Expression of the glioma-associated oncogene homolog 1 (gli1 in advanced serous ovarian cancer is associated with unfavorable overall survival.

    Alessandra Ciucci

    Full Text Available Recent evidence links aberrant activation of Hedgehog (Hh signaling with the pathogenesis of several cancers including medulloblastoma, glioblastoma, melanoma as well as pancreas, colorectal, and prostate carcinomas. Here we investigated the role of the transcription factor Gli1 in ovarian cancer. To this end, the expression profile of Gli1 was examined in normal ovaries, ovarian tumors, and ovarian cancer cell lines, and the in vitro effects of a specific Hh-pathway blocker, KAAD-cyclopamine, or a specific Gli1 inhibitor (GANT58 on cell proliferation and on Hh target gene expression were also assessed. Results obtained showed that epithelial cells in ovarian cancer tissue express significantly higher levels of nuclear Gli1 than in normal ovarian tissue, where the protein was almost undetectable. In addition, multivariate analysis showed that nuclear Gli1 was independently associated to poor survival in advanced serous ovarian cancer patients (HR = 2.2, 95%CI 1.0-5.1, p = 0.04. In vitro experiments demonstrated Gli1 expression in the three ovarian carcinoma cell lines tested, A2780, SKOV-3 and OVCAR-3. Remarkably, although KAAD-cyclopamine led to decreased cell proliferation, this treatment did not inhibit hedgehog target gene expression in any of the three ovarian cancer cell lines, suggesting that the inhibition of cell proliferation was a nonspecific or toxic effect. In line with these data, no differences on cell proliferation were observed when cell lines were treated with GANT58. Overall, our clinical data support the role of Gli1 as a prognostic marker in advanced serous ovarian cancer and as a possible therapeutic target in this disease. However, our in vitro findings draw attention to the need for selection of appropriate experimental models that accurately represent human tumor for testing future therapies involving Hh pathway inhibitors.

  14. Pre-synaptic glycine GlyT1 transporter--NMDA receptor interaction: relevance to NMDA autoreceptor activation in the presence of Mg2+ ions.

    Musante, Veronica; Summa, Maria; Cunha, Rodrigo A; Raiteri, Maurizio; Pittaluga, Anna

    2011-05-01

    Rat hippocampal glutamatergic terminals possess NMDA autoreceptors whose activation by low micromolar NMDA elicits glutamate exocytosis in the presence of physiological Mg(2+) (1.2 mM), the release of glutamate being significantly reduced when compared to that in Mg(2+)-free condition. Both glutamate and glycine were required to evoke glutamate exocytosis in 1.2 mM Mg(2+), while dizocilpine, cis-4-[phosphomethyl]-piperidine-2-carboxylic acid and 7-Cl-kynurenic acid prevented it, indicating that occupation of both agonist sites is needed for receptor activation. D-serine mimicked glycine but also inhibited the NMDA/glycine-induced release of [(3H]D-aspartate, thus behaving as a partial agonist. The NMDA/glycine-induced release in 1.2 mM Mg(2+) strictly depended on glycine uptake through the glycine transporter type 1 (GlyT1), because the GlyT1 blocker N-[3-(4'-fluorophenyl)-3-(4'-phenylphenoxy)propyl])sarcosine hydrochloride, but not the GlyT2 blocker Org 25534, prevented it. Accordingly, [(3)H]glycine was taken up during superfusion, while lowering the external concentration of Na(+), the monovalent cation co-transported with glycine by GlyT1, abrogated the NMDA-induced effect. Western blot analysis of subsynaptic fractions confirms that GlyT1 and NMDA autoreceptors co-localize at the pre-synaptic level, where GluN3A subunits immunoreactivity was also recovered. It is proposed that GlyT1s coexist with NMDA autoreceptors on rat hippocampal glutamatergic terminals and that glycine taken up by GlyT1 may permit physiological activation of NMDA pre-synaptic autoreceptors. © 2011 The Authors. Journal of Neurochemistry © 2011 International Society for Neurochemistry.

  15. X-linked Alport syndrome associated with a synonymous p.Gly292Gly mutation alters the splicing donor site of the type IV collagen alpha chain 5 gene.

    Fu, Xue Jun; Nozu, Kandai; Eguchi, Aya; Nozu, Yoshimi; Morisada, Naoya; Shono, Akemi; Taniguchi-Ikeda, Mariko; Shima, Yuko; Nakanishi, Koichi; Vorechovsky, Igor; Iijima, Kazumoto

    2016-10-01

    X-linked Alport syndrome (XLAS) is a progressive hereditary nephropathy caused by mutations in the type IV collagen alpha chain 5 gene (COL4A5). Although many COL4A5 mutations have previously been identified, pathogenic synonymous mutations have not yet been described. A family with XLAS underwent mutational analyses of COL4A5 by PCR and direct sequencing, as well as transcript analysis of potential splice site mutations. In silico analysis was also conducted to predict the disruption of splicing factor binding sites. Immunohistochemistry (IHC) of kidney biopsies was used to detect α2 and α5 chain expression. We identified a hemizygous point mutation, c.876A>T, in exon 15 of COL4A5 in the proband and his brother, which is predicted to result in a synonymous amino acid change, p.(Gly292Gly). Transcript analysis showed that this mutation potentially altered splicing because it disrupted the splicing factor binding site. The kidney biopsy of the proband showed lamellation of the glomerular basement membrane (GBM), while IHC revealed negative α5(IV) staining in the GBM and Bowman's capsule, which is typical of XLAS. This is the first report of a synonymous COL4A5 substitution being responsible for XLAS. Our findings suggest that transcript analysis should be conducted for the future correct assessment of silent mutations.

  16. A novel hemoglobin variant found on the α1 chain: Hb KSVGH (HBA1: p.Lys57_Gly58insSerHisGlySerAlaGlnValLys).

    Wang, Mei-Chun; Tsai, Kuo-Wang; Chu, Chih-Hsun; Yu, Ming-Sun; Lam, Hing-Chung

    2015-01-01

    Glycosylated hemoglobin (Hb A1C) is a crucial indicator for the long-term control and the diagnosis of diabetes. However, the presence of hemoglobin (Hb) variants may affect the measured value of Hb A1C and result in an abnormal graph trend and inconsistency between the clinical blood sugar test and Hb A1C values. In this study, laboratory data of 41,267 patients with diabetes were collected. The Hb A1C levels and the graph results were examined. We identified 74 cases containing abnormal Hb A1C graph trends. The conducted blood cell counts and capillary Hb electrophoresis were used to analyze Hb variants. We also determined gene variation for the Hb variants by a sequence approach. Fifteen different types of Hb variants were identified in this study. Among these, we found a novel variant in which the α1 subunit of Hb showed an insertion of 24 nucleotides (nts) between the 56th and 57th residues. We named this novel variant Hb Kaohsiung Veterans General Hospital (Hb KSVGH) (HBA1: p.Lys57_Gly58insSerHisGlySerAlaGlnValLys).

  17. IL FILM A LEZIONE DI ITALIANO L2: GLI ESAMI AL CINEMA

    Michela Albizzati

    2014-02-01

    Full Text Available Può il film rivelarsi uno strumento utile a lezione di italiano L2? Quali sono i limiti ed i vantaggi dell’utilizzo di materiale audiovisivo autentico in aula? Come strutturare un’attività basata sulla visione di un film italiano in lingua originale? Quali strategie mette in atto l’apprendente per comprendere un film nella L2? “Il film a lezione di italiano L2: gli esami al cinema” cerca di rispondere a queste domande attraverso una riflessione sull’utilità del cinema a lezione e proponendo un’attività didattica basata sulla visione di sequenze tratte da film italiani. Considerando la pellicola come documento di civiltà, veicolo di una lingua viva e “in situazione” ed esperienza di apprendimento plurisensoriale, l’articolo mette in luce come l’apprendimento dell’italiano L2 possa trarre beneficio dall’utilizzo in aula di materiali audiovisivi autentici quali i film, se opportunamente selezionati e didattizzati. Dalla riflessione alla pratica: dopo aver identificato obiettivi, destinatari ed i loro bisogni linguistico-comunicativi, viene presentata un’attività basata sulla didattizzazione di tre sequenze che hanno come tema centrale l’esame universitario orale. Ponendo i bisogni linguistico-comunicativi dei discenti al centro si scoprirà che, se opportunamente didattizzato, un film o parte di esso può trasformarsi in materiale didattico estremamente utile ai fini dell’apprendimento di una L2, nonché fonte inesauribile di spunti a partire dai quali l’insegnante può creare attività didattiche originali e stimolanti non solo linguistiche ma anche interculturali. Movies during italian l2 lessons: exams at the cinemaAre movies useful tools during Italian L2 lessons? What are the limits and advantages of using authentic audio-visual materials in the classroom? How can an activity based on viewing an Italian film in the original language be structured? What strategies do learners use to understand movies in

  18. Molecular Analysis of Gli3, Ihh, Rab23, and Jag1 in a Rabbit Model of Craniosynostosis: Likely Exclusion as the Loci of Origin.

    Gilbert, James R; Taylor, Gwen M; Losee, Joseph E; Mooney, Mark P; Cooper, Gregory M

    2018-03-01

    Craniosynostosis (CS) involves the premature fusion of one or more cranial sutures. The etiology of CS is complex and mutations in more than 50 distinct genes have been causally linked to the disorder. Many of the genes that have been associated with CS in humans play an essential role in tissue patterning and early craniofacial development. Among these genes are members of the Hedgehog (HH) and Notch signal transduction pathways, including the GLI family member Gli3, Indian Hedgehog ( Ihh), the RAS oncogene family member Rab23, and the Notch ligand JAGGED1 ( Jag1). We have previously described a colony of rabbits with a heritable pattern of coronal suture synostosis, although the genetic basis for synostosis within this model remains unknown. The present study was performed to determine if coding errors in Gli3, Ihh, Rab23, or Jag1 could be causally linked to craniosynostosis in this unique animal model. Sequencing of cDNA templates was performed using samples obtained from wild-type and craniosynostotic rabbits. Several nucleotide polymorphisms were identified in Gli3, Ihh, and Rab23, although these variants failed to segregate by phenotype. No nucleotide polymorphisms were identified in Jag1. These data indicate that the causal locus for heritable craniosynostosis in this rabbit model is not located within the protein coding regions of Gli3, Ihh, Rab23, or Jag1.

  19. Modulation of mitochondrial activity in HaCaT keratinocytes by the cell penetrating peptide Z-Gly-RGD(DPhe)-mitoparan.

    Richardson, Adam; Muir, Lewis; Mousdell, Sasha; Sexton, Darren; Jones, Sarah; Howl, John; Ross, Kehinde

    2018-01-30

    Biologically active cell penetrating peptides (CPPs) are an emerging class of therapeutic agent. The wasp venom peptide mastoparan is an established CPP that modulates mitochondrial activity and triggers caspase-dependent apoptosis in cancer cells, as does the mastoparan analogue mitoparan (mitP). Mitochondrial depolarisation and activation of the caspase cascade also underpins the action of dithranol, a topical agent for treatment of psoriasis. The effects of a potent mitP analogue on mitochondrial activity were therefore examined to assess its potential as a novel approach for targeting mitochondria for the treatment of psoriasis. In HaCaT keratinocytes treated with the mitP analogue Z-Gly-RGD(DPhe)-mitP for 24 h, a dose-dependent loss of mitochondrial activity was observed using the methyl-thiazolyl-tetrazolium (MTT) assay. At 10 μmol L -1 , MTT activity was less than 30% that observed in untreated cells. Staining with the cationic dye JC-1 suggested that Z-Gly-RGD(DPhe)-mitP also dissipated the mitochondrial membrane potential, with a threefold increase in mitochondrial depolarisation levels. However, caspase activity appeared to be reduced by 24 h exposure to Z-Gly-RGD(DPhe)-mitP treatment. Furthermore, Z-Gly-RGD(DPhe)-mitP treatment had little effect on overall cell viability. Our findings suggest Z-Gly-RGD(DPhe)-mitP promotes the loss of mitochondrial activity but does not appear to evoke apoptosis in HaCaT keratinocytes.

  20. The recent multi-ethnic global lung initiative 2012 (GLI2012) reference values don't reflect contemporary adult's North African spirometry.

    Ben Saad, Helmi; El Attar, Mohamed Nour; Hadj Mabrouk, Khaoula; Ben Abdelaziz, Ahmed; Abdelghani, Ahmed; Bousarssar, Mohamed; Limam, Khélifa; Maatoug, Chiraz; Bouslah, Hmida; Charrada, Ameur; Rouatbi, Sonia

    2013-12-01

    The applicability of the recent multi-ethnic reference equations derived by the ERS Global Lung Initiative (ERS/GLI) in interpreting spirometry data in North African adult subjects has not been studied. To ascertain how well the recent ERS/GLI reference equations fit contemporary adult Tunisian spirometric data. Spirometric data were recorded from 1192 consecutive spirometry procedures in adults aged 18-60 years. Reference values and lower limits of normality (LLN) were calculated using the local and the ERS/GLI reference equations. Applied definitions: large airway obstructive ventilatory defect (LAOVD): FEV1/FVC contemporary Tunisian spirometry. Using Tunisian reference equations, 71.31%, 6.71% and 19.04% of spirometry records were interpreted as normal, and as having, LAOVD and TRVD, respectively. Using the ERS/GLI reference equations, these figures were respectively, 85.82%, 4.19% and 8.39%. The mean ± SD Z-scores for the contemporary healthy North African subject's data were -0.55 ± 0.87 for FEV1, -0.62 ± 0.86 for FVC and 0.10 ± 0.73 for FEV1/FVC. The present study don't recommend the use of the recent ERS/GLI reference equations to interpret spirometry in North African adult population. Copyright © 2013 Elsevier Ltd. All rights reserved.

  1. Sonic hedgehog (SHH) and glioblastoma-2 (Gli-2) expressions are associated with poor jaundice-free survival in biliary atresia.

    Jung, Hae Yoen; Jing, Jin; Lee, Kyoung Bun; Jang, Ja-June

    2015-03-01

    Biliary atresia (BA) causes biliary obstruction in neonates. Although the Kasai operation can successfully treat certain BA cases, many patients exhibit recurrent jaundice and secondary biliary cirrhosis requiring liver transplantation. Consequently, studies of the prognostic factors of the Kasai operation are needed. Accordingly, sonic hedgehog (SHH) pathway expression at the extrahepatic bile duct (EHBD), an important bile duct repair mechanism, will be investigated via immunohistochemistry in patients with BA to examine the association with post-Kasai operation prognosis. Fifty-seven EHBD specimens were obtained during Kasai operations from 1992 to 2009. The SHH, patched (PTCH), and glioblastoma-2 (Gli-2) immunohistochemical staining results were analyzed quantitatively. Overall, 57.9% of patients had bile flow normalization after the Kasai operation; 43.1% did not. High preoperative serum total bilirubin, direct bilirubin, and aspartate aminotransferase levels were associated with sustained jaundice post-Kasai operation, as was an age ≥65days at the time of surgery (all pjaundice relapse. Strong Gli-2 and SHH expression in the EHBD might be a poor prognostic factor in Kasai operation-treated patients with BA. Copyright © 2015 Elsevier Inc. All rights reserved.

  2. Diarylheptanoids suppress proliferation of pancreatic cancer PANC-1 cells through modulating shh-Gli-FoxM1 pathway.

    Dong, Guang-Zhi; Jeong, Ji Hye; Lee, Yu-Ih; Lee, So Yoon; Zhao, Hui-Yuan; Jeon, Raok; Lee, Hwa Jin; Ryu, Jae-Ha

    2017-04-01

    Pancreatic cancer is one of the leading causes of cancer, and it has the lowest 5-year survival rates. It is necessary to develop more potent anti-pancreatic cancer drugs to overcome the fast metastasis and resistance to surgery, radiotherapy, chemotherapy, and combinations of these. We have identified several diarylheptanoids as anti-pancreatic cancer agents from Alpinia officinarum (lesser galangal) and Alnus japonica. These diarylheptanoids suppressed cell proliferation and induced the cell cycle arrest of pancreatic cancer cells (PANC-1). Among them, the most potent compounds 1 and 7 inhibited the shh-Gli-FoxM1 pathway and their target gene expression in PANC-1 cells. Furthermore, they suppressed the expression of the cell cycle associated genes that were rescued by the overexpression of exogenous FoxM1. Taken together, (E)-7-(4-hydroxy-3-methoxyphenyl)-1-phenylhept-4-en-3-one (1) from Alpinia officinarum (lesser galangal) and platyphyllenone (7) from Alnus japonica inhibit PANC-1 cell proliferation by suppressing the shh-Gli-FoxM1 pathway, and they can be potential candidates for anti-pancreatic cancer drug development.

  3. Analysis of PGC-1α variants Gly482Ser and Thr612Met concerning their PPARγ2-coactivation function

    Nitz, Inke; Ewert, Agnes; Klapper, Maja; Doering, Frank

    2007-01-01

    Peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) is a cofactor involved in adaptive thermogenesis, fatty acid oxidation, and gluconeogenesis. Dysfunctions of this protein are likely to contribute to the development of obesity and the metabolic syndrome. This is in part but not definitely confirmed by results of population studies. The aim of this study was to investigate if common genetic variants rs8192678 (Gly482Ser) and rs3736265 (Thr612Met) in the PGC-1α gene lead to a functional consequence in cofactor activity using peroxisome proliferator-activated receptor-γ 2 (PPARγ2) as interacting transcription factor. Reporter gene assays in HepG2 cells with wildtype and mutant proteins of both PGC1α and PPARγ2 (Pro12Ala, rs1801282) using the acyl-CoA-binding protein (ACBP) promoter showed no difference in coactivator activity. This is First study implicating that the Gly482Ser and Thr612Met polymorphisms in PGC-1α and Pro12Ala polymorphism in PPARγ2 do not affect the functional integrity of these proteins

  4. A Gly482Ser missense mutation in the peroxisome proliferator-activated receptor gamma coactivator-1 is associated with altered lipid oxidation and early insulin secretion in Pima Indians

    Muller, Yunhua Li; Bogardus, Clifton; Pedersen, Oluf

    2003-01-01

    of the PGC-1 gene predicts a glycine to serine substitution at amino acid 482 and has been associated with type 2 diabetes in a Danish population. In this study, we examined whether this Gly482Ser polymorphism is associated with type 2 diabetes or obesity, or metabolic predictors of these diseases, in Pima...... Indians. There was no association of the Gly482Ser polymorphism with either type 2 diabetes or BMI (n = 984). However, among nondiabetic Pima Indians (n = 183-201), those with the Gly/Gly genotype had a lower mean insulin secretory response to intravenous and oral glucose and a lower mean rate of lipid...

  5. The Bridge Impacts: Unsustainable Undergrowth Gli impatti del Ponte: sottosviluppo insostenibile

    Alberto Ziparo

    2011-11-01

    " contenuta nel progetto '80, alle posizioni critiche che, rispetto a tali concetti, sono maturate negli studi relativi alle più recenti versioni del progetto. Nella seconda parte si descrivono gli elementi principali del "progetto di attraversamento aereo dello Stretto di Messina" presentato dalla società dello Stretto al governo il 31/12/02 (progetto preliminare, che ha registrato il parere positivo con prescrizioni del CIPE nel 2005, di cui è stata di recente presentata la versione “definitiva”; e ancora note tratte dalle "Osservazioni sullo studio di impatto ambientale", avanzate per conto di Legambiente, WWF, Italia Nostra e di altre associazioni ambientaliste, da un gruppo di studio coordinato dall'autore delle note presenti, riassumendo anche recenti lavori di aggiornamento (Bettini, Guerzoni, Ziparo, 2002; Pieroni, 2000; Pieroni, Ziparo, 2010; Marino, 2011.

  6. Hedgehog signaling pathway is active in GBM with GLI1 mRNA expression showing a single continuous distribution rather than discrete high/low clusters.

    Chandra, Vikas; Das, Tapojyoti; Gulati, Puneet; Biswas, Nidhan K; Rote, Sarang; Chatterjee, Uttara; Ghosh, Samarendra N; Deb, Sumit; Saha, Suniti K; Chowdhury, Anup K; Ghosh, Subhashish; Rudin, Charles M; Mukherjee, Ankur; Basu, Analabha; Dhara, Surajit

    2015-01-01

    Hedgehog (Hh) signaling pathway is a valid therapeutic target in a wide range of malignancies. We focus here on glioblastoma multiforme (GBM), a lethal malignancy of the central nervous system (CNS). By analyzing RNA-sequencing based transcriptomics data on 149 clinical cases of TCGA-GBM database we show here a strong correlation (r = 0.7) between GLI1 and PTCH1 mRNA expression--as a hallmark of the canonical Hh-pathway activity in this malignancy. GLI1 mRNA expression varied in 3 orders of magnitude among the GBM patients of the same cohort showing a single continuous distribution-unlike the discrete high/low-GLI1 mRNA expressing clusters of medulloblastoma (MB). When compared with MB as a reference, the median GLI1 mRNA expression in GBM appeared 14.8 fold lower than that of the "high-Hh" cluster of MB but 5.6 fold higher than that of the "low-Hh" cluster of MB. Next, we demonstrated statistically significant up- and down-regulation of GLI1 mRNA expressions in GBM patient-derived low-passage neurospheres in vitro by sonic hedgehog ligand-enriched conditioned media (shh-CM) and by Hh-inhibitor drug vismodegib respectively. We also showed clinically achievable dose (50 μM) of vismodegib alone to be sufficient to induce apoptosis and cell cycle arrest in these low-passage GBM neurospheres in vitro. Vismodegib showed an effect on the neurospheres, both by down-regulating GLI1 mRNA expression and by inducing apoptosis/cell cycle arrest, irrespective of their relative endogenous levels of GLI1 mRNA expression. We conclude from our study that this single continuous distribution pattern of GLI1 mRNA expression technically puts almost all GBM patients in a single group rather than discrete high- or low-clusters in terms of Hh-pathway activity. That is suggestive of therapies with Hh-pathway inhibitor drugs in this malignancy without a need for further stratification of patients on the basis of relative levels of Hh-pathway activity among them.

  7. The Gly482Ser genotype at the PPARGC1A gene and elevated blood pressure: a meta-analysis involving 13,949 individuals

    Vimaleswaran, Karani Santhanakrishnan; Luan, Jian'an; Andersen, Gitte

    2008-01-01

    The protein encoded by the PPARGC1A gene is expressed at high levels in metabolically active tissues and is involved in the control of oxidative stress via reactive oxygen species detoxification. Several recent reports suggest that the PPARGC1A Gly482Ser (rs8192678) missense polymorphism may relate...... inversely with blood pressure. We used conventional meta-analysis methods to assess the association between Gly482Ser and systolic (SBP) or diastolic blood pressures (DBP) or hypertension in 13,949 individuals from 17 studies, of which 6,042 were previously unpublished observations. The studies comprised...

  8. Evaluation of the oxidative stress modulation in Drosophila melanogaster strains deficient in endogenous antioxidants and with chronic exposure to casiopeina Cas II-gly and gamma radiation

    Jimenez V, E. R.

    2013-01-01

    The casiopeinas are a family of coordination compounds with copper metallic center that have shown to have antineoplastic activity. The experimental evidences suggest that its action mechanism is through the generation of free radicals. The casiopeina (Cas II-gly) is believed to causes oxidative damage in the mitochondria, leading to the cellular death. The present study has the purpose to evaluate the antioxidant potential of the tetrapyrroles: cupro-sodica chlorophyllin (CSC), protoporphyrin-Ix (Pp-Ix) and the bilirubin (Bili) against the oxidant action of the Cas II-gly. The present study will also contribute in the characterization of the biological activity of the Cas II-gly. For this purpose is quantifies the effect of these compounds in the enzymes activity, superoxide dismutase (Sod) and catalase (Cat) in wild Drosophila melanogaster strains Canton-S and in the deficient in Sod and Cat. Two protocols were used, in the first male of 1-24 h of age were pre-treated with 0, 0.01, 0.1 and 1 m M of Cas II-gly and later on they were treated with radiation (15 Gy), and the second 69 m M of CSC, Pp-Ix or Bili, during 8 days and later they were treated with 0.1 m M of Cas II-gly during 24 h. The enzymatic activity was measured with the detection packages of enzymes Sod and Cat of Sigma. It was found that none of the three pigments increment the Sod activity but, if they diminished that of Cat (p≤0.007). The three concentrations of Cas II-gly did not increase the Sod activity significantly, only the concentration of 0.1 m M diminishes in 5.6 U the Cat activity (p <0.03) the same as the treatment with 15 Gy of gamma rays (8 U, p <0.004). The Cas II-gly combination 0.1 m M with the pigments does not modify the Sod and Cat activity. These results suggest that the proven pigments act as antioxidants, avoiding the induction of exogenous antioxidants caused by the gamma rays or the Cas II-gly. (Author)

  9. Pituitary adenylyl cyclase activating polypeptide inhibits gli1 gene expression and proliferation in primary medulloblastoma derived tumorsphere cultures

    Dong Hongmei

    2010-12-01

    Full Text Available Abstract Background Hedgehog (HH signaling is critical for the expansion of granule neuron precursors (GNPs within the external granular layer (EGL during cerebellar development. Aberrant HH signaling within GNPs is thought to give rise to medulloblastoma (MB - the most commonly-observed form of malignant pediatric brain tumor. Evidence in both invertebrates and vertebrates indicates that cyclic AMP-dependent protein kinase A (PKA antagonizes HH signalling. Receptors specific for the neuropeptide pituitary adenylyl cyclase activating polypeptide (PACAP, gene name ADCYAP1 are expressed in GNPs. PACAP has been shown to protect GNPs from apoptosis in vitro, and to interact with HH signaling to regulate GNP proliferation. PACAP/ptch1 double mutant mice exhibit an increased incidence of MB compared to ptch1 mice, indicating that PACAP may regulate HH pathway-mediated MB pathogenesis. Methods Primary MB tumorsphere cultures were prepared from thirteen ptch1+/-/p53+/- double mutant mice and treated with the smoothened (SMO agonist purmorphamine, the SMO antagonist SANT-1, the neuropeptide PACAP, the PKA activator forskolin, and the PKA inhibitor H89. Gene expression of gli1 and [3H]-thymidine incorporation were assessed to determine drug effects on HH pathway activity and proliferation, respectively. PKA activity was determined in cell extracts by Western blotting using a phospho-PKA substrate antibody. Results Primary tumor cells cultured for 1-week under serum-free conditions grew as tumorspheres and were found to express PAC1 receptor transcripts. Gli1 gene expression was significantly reduced by SANT-1, PACAP and forskolin, but was unaffected by purmorphamine. The attenuation of gli1 gene expression by PACAP was reversed by the PKA inhibitor H89, which also blocked PKA activation. Treatment of tumorsphere cultures with PACAP, forskolin, and SANT-1 for 24 or 48 hours reduced proliferation. Conclusions Primary tumorspheres derived from ptch1+/-/p53

  10. Pituitary adenylyl cyclase activating polypeptide inhibits gli1 gene expression and proliferation in primary medulloblastoma derived tumorsphere cultures

    Cohen, Joseph R; Resnick, Daniel Z; Niewiadomski, Pawel; Dong, Hongmei; Liau, Linda M; Waschek, James A

    2010-01-01

    Hedgehog (HH) signaling is critical for the expansion of granule neuron precursors (GNPs) within the external granular layer (EGL) during cerebellar development. Aberrant HH signaling within GNPs is thought to give rise to medulloblastoma (MB) - the most commonly-observed form of malignant pediatric brain tumor. Evidence in both invertebrates and vertebrates indicates that cyclic AMP-dependent protein kinase A (PKA) antagonizes HH signalling. Receptors specific for the neuropeptide pituitary adenylyl cyclase activating polypeptide (PACAP, gene name ADCYAP1) are expressed in GNPs. PACAP has been shown to protect GNPs from apoptosis in vitro, and to interact with HH signaling to regulate GNP proliferation. PACAP/ptch1 double mutant mice exhibit an increased incidence of MB compared to ptch1 mice, indicating that PACAP may regulate HH pathway-mediated MB pathogenesis. Primary MB tumorsphere cultures were prepared from thirteen ptch1 +/- /p53 +/- double mutant mice and treated with the smoothened (SMO) agonist purmorphamine, the SMO antagonist SANT-1, the neuropeptide PACAP, the PKA activator forskolin, and the PKA inhibitor H89. Gene expression of gli1 and [ 3 H]-thymidine incorporation were assessed to determine drug effects on HH pathway activity and proliferation, respectively. PKA activity was determined in cell extracts by Western blotting using a phospho-PKA substrate antibody. Primary tumor cells cultured for 1-week under serum-free conditions grew as tumorspheres and were found to express PAC1 receptor transcripts. Gli1 gene expression was significantly reduced by SANT-1, PACAP and forskolin, but was unaffected by purmorphamine. The attenuation of gli1 gene expression by PACAP was reversed by the PKA inhibitor H89, which also blocked PKA activation. Treatment of tumorsphere cultures with PACAP, forskolin, and SANT-1 for 24 or 48 hours reduced proliferation. Primary tumorspheres derived from ptch1 +/- /p53 +/- mice exhibit constitutive HH pathway activity

  11. Severe coagulation factor VII deficiency caused by a novel homozygous mutation (p. Trp284Gly) in loop 140s.

    Hao, Xiuping; Cheng, XiaoLi; Ye, Jiajia; Wang, Yingyu; Yang, LiHong; Wang, Mingshan; Jin, Yanhui

    2016-06-01

    Congenital coagulation factor VII (FVII) deficiency is a rare disorder caused by mutation in F7 gene. Herein, we reported a patient who had unexplained hematuria and vertigo with consanguineous parents. He has been diagnosed as having FVII deficiency based on the results of reduced FVII activity (2.0%) and antigen (12.8%). The thrombin generation tests verified that the proband has obstacles in producing thrombin. Direct sequencing analysis revealed a novel homozygous missense mutation p.Trp284Gly. Also noteworthy is the fact that the mutational residue belongs to structurally conserved loop 140s, which majorly undergo rearrangement after FVII activation. Model analysis indicated that the substitution disrupts these native hydrophobic interactions, which are of great importance to the conformation in the activation domain of FVIIa.

  12. Direct Assessment of the Effect of the Gly380Arg Achondroplasia Mutation on FGFR3 Dimerization Using Quantitative Imaging FRET

    Placone, Jesse; Hristova, Kalina

    2012-01-01

    The Gly380Arg mutation in FGFR3 is the genetic cause for achondroplasia (ACH), the most common form of human dwarfism. The mutation has been proposed to increase FGFR3 dimerization, but the dimerization propensities of wild-type and mutant FGFR3 have not been compared. Here we use quantitative imaging FRET to characterize the dimerization of wild-type FGFR3 and the ACH mutant in plasma membrane-derived vesicles from HEK293T cells. We demonstrate a small, but statistically significant increase in FGFR3 dimerization due to the ACH mutation. The data are consistent with the idea that the ACH mutation causes a structural change which affects both the stability and the activity of FGFR3 dimers in the absence of ligand. PMID:23056398

  13. Direct assessment of the effect of the Gly380Arg achondroplasia mutation on FGFR3 dimerization using quantitative imaging FRET.

    Jesse Placone

    Full Text Available The Gly380Arg mutation in FGFR3 is the genetic cause for achondroplasia (ACH, the most common form of human dwarfism. The mutation has been proposed to increase FGFR3 dimerization, but the dimerization propensities of wild-type and mutant FGFR3 have not been compared. Here we use quantitative imaging FRET to characterize the dimerization of wild-type FGFR3 and the ACH mutant in plasma membrane-derived vesicles from HEK293T cells. We demonstrate a small, but statistically significant increase in FGFR3 dimerization due to the ACH mutation. The data are consistent with the idea that the ACH mutation causes a structural change which affects both the stability and the activity of FGFR3 dimers in the absence of ligand.

  14. Microbial expression of proteins containing long repetitive Arg-Gly-Asp cell adhesive motifs created by overlap elongation PCR

    Kurihara, Hiroyuki; Shinkai, Masashige; Nagamune, Teruyuki

    2004-01-01

    We developed a novel method for creating repetitive DNA libraries using overlap elongation PCR, and prepared a DNA library encoding repetitive Arg-Gly-Asp (RGD) cell adhesive motifs. We obtained various length DNAs encoding repetitive RGD from a short monomer DNA (18 bp) after a thermal cyclic reaction without a DNA template for amplification, and isolated DNAs encoding 2, 21, and 43 repeats of the RGD motif. We cloned these DNAs into a protein expression vector and overexpressed them as thioredoxin fusion proteins: RGD2, RGD21, and RGD43, respectively. The solubility of RGD43 in water was low and it formed a fibrous precipitate in water. Scanning electron microscopy revealed that RGD43 formed a branched 3D-network structure in the solid state. To evaluate the function of the cell adhesive motifs in RGD43, mouse fibroblast cells were cultivated on the RGD43 scaffold. The fibroblast cells adhered to the RGD43 scaffold and extended long filopodia

  15. Gli effetti socio-economici e spaziali del servizio ferroviario dell’Alta Velocità: due casi a confronto

    Loredana Consuelo Travascio

    2008-05-01

    Full Text Available L’articolo fornisce una lettura degli effetti socio-economici e spaziali generati dalla messa in esercizio di una rete ferroviaria ad AV sull’organizzazione territoriale. Esso si articola in due parti. Nella prima, sono analizzati gli effetti che in letteratura sono attribuiti alle infrastrutture di trasporto, focalizzandosi sulle ripercussioni che la messa in esercizio dell’AV ha generato sulla crescita demografica e sulla localizzazione delle attività economiche attorno alle stazioni, nonché sulla domanda di trasporto e sulla riconfigurazione dei flussi. Nella seconda parte si passa allo studio di due casi significativi: il Giappone, dove l’introduzione del servizio ha modificato la struttura socio-economica del territorio; la Spagna, dove ha favorito la comparsa di nuove tipologie di utenti e la ridefinizione delle relazioni tra le varie città servite dall’infrastruttura.

  16. Proteolysis of His-Phe-Arg-Trp-Pro-Gly-Pro in the blood and brain of rats in vivo.

    Shevchenko, K V; Nagaev, I Yu; Babakov, V N; Andreeva, L A; Shevchenko, V P; Radilov, A S; Myasoedov, N F

    2015-01-01

    The kinetics of the content of His-Phe-Arg-Trp-Pro-Gly-Pro (ACTH (6-9)PGP) and its hydrolysis products in the blood and brain of rats in the case of intranasal administration and intravenous injection of tritiated ACTH(6-9)PGP was studied. The parameters of bioavailability of ACTH(6-9)PGP administered intranasally were higher, indicating certain prospects in the intranasal application in clinical practice. We also found that the factor that determines ACTH(6-9)PGP proteolysis in experiments both in vivo and in vitro is aminopeptidases. The main products of ACTH(6-9)PGP during its metabolism in rats are short peptides and amino acids.

  17. Dopamine D3 receptor Ser9Gly variant is associated with impulse control disorders in Parkinson's disease patients.

    Krishnamoorthy, Soumya; Rajan, Roopa; Banerjee, Moinak; Kumar, Hardeep; Sarma, Gangadhara; Krishnan, Syam; Sarma, Sankara; Kishore, Asha

    2016-09-01

    Impulse control disorders (ICD) are reported to occur at variable frequencies in different ethnic groups. Genetic vulnerability is suspected to underlie the individual risk for ICD. We investigated whether the allelic variants of dopamine (DRD3), glutamate (GRIN2B) and serotonin (HTR2A) receptors are linked to ICD in Indian Parkinson's disease (PD) patients. We conducted a prospective, case-control study which included PD patients (70 with ICD, 100 without ICD categorized after direct psychiatric interview of patient and caregiver) and 285 healthy controls. Single nucleotide polymorphism (SNP) variants of DRD3 p.S9G (rs6280), GRIN2B c.2664C>T (rs1806201) and HTR2A c.102T>C (rs6313) were genotyped. Multivariate regression analysis revealed that DRD3 p.Ser9Gly (rs6280) heterozygous variant CT (OR = 2.22, 95% CI: 1.03-4.86, p = 0.041), higher daily Levodopa equivalent doses (LED) of drugs (for 100 mg LED, OR = 1.14, 95% CI: 1.01-1.29, p = 0.041), current dopamine agonist but not Levodopa use (OR = 2.16, 95% CI: 1.03-4.55, p = 0.042) and age of onset of motor symptoms under 50 years (OR 2.09, 95% CI: 1.05-4.18, p = 0.035) were independently associated with ICD. DRD3 p.Ser9Gly (rs6280) CT genotype is associated with ICD in Indian PD patients and this association is novel. Enhanced D3 receptor affinity due to gain-of-function conferred by the glycine residues could impair reward-risk assessment in the mesolimbic system and contribute to development of impulsive behaviour, in carriers of this genotype. Copyright © 2016. Published by Elsevier Ltd.

  18. The functional DRD3 Ser9Gly polymorphism (rs6280) is pleiotropic, affecting reward as well as movement.

    Savitz, Jonathan; Hodgkinson, Colin A; Martin-Soelch, Chantal; Shen, Pei-Hong; Szczepanik, Joanna; Nugent, Allison; Herscovitch, Peter; Grace, Anthony A; Goldman, David; Drevets, Wayne C

    2013-01-01

    Abnormalities of motivation and behavior in the context of reward are a fundamental component of addiction and mood disorders. Here we test the effect of a functional missense mutation in the dopamine 3 receptor (DRD3) gene (ser9gly, rs6280) on reward-associated dopamine (DA) release in the striatum. Twenty-six healthy controls (HCs) and 10 unmedicated subjects with major depressive disorder (MDD) completed two positron emission tomography (PET) scans with [(11)C]raclopride using the bolus plus constant infusion method. On one occasion subjects completed a sensorimotor task (control condition) and on another occasion subjects completed a gambling task (reward condition). A linear regression analysis controlling for age, sex, diagnosis, and self-reported anhedonia indicated that during receipt of unpredictable monetary reward the glycine allele was associated with a greater reduction in D2/3 receptor binding (i.e., increased reward-related DA release) in the middle (anterior) caudate (p<0.01) and the ventral striatum (p<0.05). The possible functional effect of the ser9gly polymorphism on DA release is consistent with previous work demonstrating that the glycine allele yields D3 autoreceptors that have a higher affinity for DA and display more robust intracellular signaling. Preclinical evidence indicates that chronic stress and aversive stimulation induce activation of the DA system, raising the possibility that the glycine allele, by virtue of its facilitatory effect on striatal DA release, increases susceptibility to hyperdopaminergic responses that have previously been associated with stress, addiction, and psychosis.

  19. The functional DRD3 Ser9Gly polymorphism (rs6280 is pleiotropic, affecting reward as well as movement.

    Jonathan Savitz

    Full Text Available Abnormalities of motivation and behavior in the context of reward are a fundamental component of addiction and mood disorders. Here we test the effect of a functional missense mutation in the dopamine 3 receptor (DRD3 gene (ser9gly, rs6280 on reward-associated dopamine (DA release in the striatum. Twenty-six healthy controls (HCs and 10 unmedicated subjects with major depressive disorder (MDD completed two positron emission tomography (PET scans with [(11C]raclopride using the bolus plus constant infusion method. On one occasion subjects completed a sensorimotor task (control condition and on another occasion subjects completed a gambling task (reward condition. A linear regression analysis controlling for age, sex, diagnosis, and self-reported anhedonia indicated that during receipt of unpredictable monetary reward the glycine allele was associated with a greater reduction in D2/3 receptor binding (i.e., increased reward-related DA release in the middle (anterior caudate (p<0.01 and the ventral striatum (p<0.05. The possible functional effect of the ser9gly polymorphism on DA release is consistent with previous work demonstrating that the glycine allele yields D3 autoreceptors that have a higher affinity for DA and display more robust intracellular signaling. Preclinical evidence indicates that chronic stress and aversive stimulation induce activation of the DA system, raising the possibility that the glycine allele, by virtue of its facilitatory effect on striatal DA release, increases susceptibility to hyperdopaminergic responses that have previously been associated with stress, addiction, and psychosis.

  20. Juvenil hæmokromatose forårsaget af homozygot Gly320Val-mutation på hæmojuvelingenet

    Berg, Line Brunemark; Milman, Nils Thorm; Friis-Hansen, Lennart

    2013-01-01

    Juvenile haemochromatosis caused by a homozygous Gly320Val mutation in the haemojuvelin (HJV) gene was diagnosed in a 12-year-old Danish girl and her 10-year-old sister. Both appeared healthy without clinical or biochemical signs of organ damage. They had iron overload (plasma transferrin saturat...

  1. Immunity to endotoxin and Asp299Gly polymorphism of TLR-4 in adult patients with early and late onset of asthma

    Yu. A. Bisyuk

    2015-06-01

    Full Text Available Aim. The gene polymorphism of Asp299Gly TLR-4 may be associated with the risk of asthma development. Methods and results. The gene polymorphism of TLR-4 (Asp299Gly receptor has been researched in 262 early-onset and in 69 late-onset asthma patients. The state of anti-endotoxin immunity was assessed by determination of specific antibodies to the endotoxin of A, M, G classes and sCD14 by ELISA. The polymorphism was analyzed by the allele-specific polymerase chain reaction with electrophoretic detection. It was estimated that the risk of early-onset asthma in the population of Crimea is associated with genotypes AG and GG (Asp299Gly of TLR-4. There were increased levels of anti-endotoxin IgM and decreased of sIgA in patients with late-onset asthma and AA genotype as compared to other genotypes. Conclusion. The gene polymorphism of Asp299Gly TLR-4 is associated with the risk of early-onset asthma development in Crimea population.

  2. BNNTs under the influence of external electric field as potential new drug delivery vehicle of Glu, Lys, Gly and Ser amino acids: A first-principles study

    Farmanzadeh, Davood; Ghazanfary, Samereh

    2014-01-01

    Graphical abstract: - Highlights: • Solvation energies show that the BNNTs/amino acids complex stabilizes in presence of solvent. • The adsorption process is sensitive to the external electric field. • The electric field is a suitable method for adsorption and storage of amino acids on BNNTs. - Abstract: The interaction of Glu (Glutamic acid), Lys (Lysine), Gly (Glycine) and Ser (Serine) amino acids with different polarities and (9, 0) zigzag single-wall boron nitride nanotubes (BNNTs) with various lengths in the presence and absence of external electric field (EF) in gas and solvent phases, are studied using density functional theory. It is found that interaction of Glu, Lys, Gly and Ser amino acids with BNNTs in both phases is energetically favorable. From solvation energy calculations, it can be seen that the BNNTs/amino acid complex dissolution in water is spontaneous. The adsorption energies and quantum molecular descriptors changed in the presence of external EF. Therefore, the study of BNNTs/amino acid complex under influence of external electric field is very important in proposing or designing new drug delivery systems in the presence of external EF. Results indicate that Glu, Lys, Gly and Ser amino acids can be adsorbed considerably on the BNNTs in the existence of external electric field. Our results showed that the BNNTs can act as a suitable drug delivery vehicle of Glu, Lys, Gly and Ser amino acids within biological systems and strength of adsorption and rate of drug release can be controlled by the external EF

  3. Association Analysis of Arg16Gly Polymorphism of the Beta2-adreneric Receptor Gene in Offspring from Hypertensive and Normotensive Families

    Jindra, A.; Horký, K.; Peleška, Jan; Jáchymová, M.; Bultas, J.; Umnerová, V.; Heller, S.; Hlubocká, Z.

    2002-01-01

    Roč. 11, č. 4 (2002), s. 213-217 ISSN 0803-7051 R&D Projects: GA MZd NA5615 Keywords : Arg16Gly polymorphism of the beta2-adrenergic receptor gene * normotensive offspring * essential hypertension Subject RIV: BB - Applied Statistics, Operational Research Impact factor: 1.344, year: 2002

  4. Atomic-resolution structure of the CAP-Gly domain of dynactin on polymeric microtubules determined by magic angle spinning NMR spectroscopy.

    Yan, Si; Guo, Changmiao; Hou, Guangjin; Zhang, Huilan; Lu, Xingyu; Williams, John Charles; Polenova, Tatyana

    2015-11-24

    Microtubules and their associated proteins perform a broad array of essential physiological functions, including mitosis, polarization and differentiation, cell migration, and vesicle and organelle transport. As such, they have been extensively studied at multiple levels of resolution (e.g., from structural biology to cell biology). Despite these efforts, there remain significant gaps in our knowledge concerning how microtubule-binding proteins bind to microtubules, how dynamics connect different conformational states, and how these interactions and dynamics affect cellular processes. Structures of microtubule-associated proteins assembled on polymeric microtubules are not known at atomic resolution. Here, we report a structure of the cytoskeleton-associated protein glycine-rich (CAP-Gly) domain of dynactin motor on polymeric microtubules, solved by magic angle spinning NMR spectroscopy. We present the intermolecular interface of CAP-Gly with microtubules, derived by recording direct dipolar contacts between CAP-Gly and tubulin using double rotational echo double resonance (dREDOR)-filtered experiments. Our results indicate that the structure adopted by CAP-Gly varies, particularly around its loop regions, permitting its interaction with multiple binding partners and with the microtubules. To our knowledge, this study reports the first atomic-resolution structure of a microtubule-associated protein on polymeric microtubules. Our approach lays the foundation for atomic-resolution structural analysis of other microtubule-associated motors.

  5. Evaluation of the oxidative stress modulation in Drosophila melanogaster strains deficient in endogenous antioxidants and with chronic exposure to casiopeina Cas II-gly and gamma radiation; Evaluacion de la modulacion del estres oxidante en cepas de Drosophila melanogaster deficientes en antioxidantes endogenos y con exposicion cronica a casiopeina CII-gly y radiacion gamma

    Jimenez V, E. R.

    2013-07-01

    The casiopeinas are a family of coordination compounds with copper metallic center that have shown to have antineoplastic activity. The experimental evidences suggest that its action mechanism is through the generation of free radicals. The casiopeina (Cas II-gly) is believed to causes oxidative damage in the mitochondria, leading to the cellular death. The present study has the purpose to evaluate the antioxidant potential of the tetrapyrroles: cupro-sodica chlorophyllin (CSC), protoporphyrin-Ix (Pp-Ix) and the bilirubin (Bili) against the oxidant action of the Cas II-gly. The present study will also contribute in the characterization of the biological activity of the Cas II-gly. For this purpose is quantifies the effect of these compounds in the enzymes activity, superoxide dismutase (Sod) and catalase (Cat) in wild Drosophila melanogaster strains Canton-S and in the deficient in Sod and Cat. Two protocols were used, in the first male of 1-24 h of age were pre-treated with 0, 0.01, 0.1 and 1 m M of Cas II-gly and later on they were treated with radiation (15 Gy), and the second 69 m M of CSC, Pp-Ix or Bili, during 8 days and later they were treated with 0.1 m M of Cas II-gly during 24 h. The enzymatic activity was measured with the detection packages of enzymes Sod and Cat of Sigma. It was found that none of the three pigments increment the Sod activity but, if they diminished that of Cat (p≤0.007). The three concentrations of Cas II-gly did not increase the Sod activity significantly, only the concentration of 0.1 m M diminishes in 5.6 U the Cat activity (p <0.03) the same as the treatment with 15 Gy of gamma rays (8 U, p <0.004). The Cas II-gly combination 0.1 m M with the pigments does not modify the Sod and Cat activity. These results suggest that the proven pigments act as antioxidants, avoiding the induction of exogenous antioxidants caused by the gamma rays or the Cas II-gly. (Author)

  6. Structural requirements of acylated Gly-l-Ala-d-Glu analogs for activation of the innate immune receptor NOD2.

    Gobec, Martina; Mlinarič-Raščan, Irena; Dolenc, Marija Sollner; Jakopin, Žiga

    2016-06-30

    The fragment of bacterial peptidoglycan muramyl dipeptide (MDP) has long been known for its adjuvant activity, however the underlying mechanism of this action has only recently been elucidated. It is ascribed to its agonist action on the nucleotide-binding oligomerization domain-containing protein 2 (NOD2). In spite of the pressing need for novel adjuvants for human use, this discovery is hampered, by not knowing the structural requirements underlying the immunostimulatory activity. We have investigated how minor modifications of hit compound acyl Gly-L-Ala-D-Glu derivative I modulate the molecular recognition by NOD2. A series of novel desmuramyldipeptides has been designed and synthesized leading to the identification of compound 16, in which the sugar moiety is replaced by a 6-phenylindole moiety, that exhibits the strongest NOD2 activation to date sans the carbohydrate moiety. The results have enabled a deeper understanding of the structural requirements of desmuramylpeptides for NOD2 activation. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  7. Sox2, Tlx, Gli3, and Her9 converge on Rx2 to define retinal stem cells in vivo.

    Reinhardt, Robert; Centanin, Lázaro; Tavhelidse, Tinatini; Inoue, Daigo; Wittbrodt, Beate; Concordet, Jean-Paul; Martinez-Morales, Juan Ramón; Wittbrodt, Joachim

    2015-06-03

    Transcriptional networks defining stemness in adult neural stem cells (NSCs) are largely unknown. We used the proximal cis-regulatory element (pCRE) of the retina-specific homeobox gene 2 (rx2) to address such a network. Lineage analysis in the fish retina identified rx2 as marker for multipotent NSCs. rx2-positive cells located in the peripheral ciliary marginal zone behave as stem cells for the neuroretina, or the retinal pigmented epithelium. We identified upstream regulators of rx2 interrogating the rx2 pCRE in a trans-regulation screen and focused on four TFs (Sox2, Tlx, Gli3, and Her9) activating or repressing rx2 expression. We demonstrated direct interaction of the rx2 pCRE with the four factors in vitro and in vivo. By conditional mosaic gain- and loss-of-function analyses, we validated the activity of those factors on regulating rx2 transcription and consequently modulating neuroretinal and RPE stem cell features. This becomes obvious by the rx2-mutant phenotypes that together with the data presented above identify rx2 as a transcriptional hub balancing stemness of neuroretinal and RPE stem cells in the adult fish retina. © 2015 The Authors. Published under the terms of the CC BY NC ND 4.0 license.

  8. GIANFRANCO BETTETINI, Il timpano dell’occhio. Gli intrecci e i giochi dei suoni e delle immagini, Milano, Bompiani, 2009

    Alessandro Bratus

    2013-12-01

    Full Text Available Il timpano dell’occhio di Gianfranco Bettetini è uno studio dedicato all’analisi semiotica dei testi audiovisivi. Nel volume l’autore si propone di indagare gli effetti che l’azione «strutturalmente sinergica e compatta» (p. 9 di suono e immagine produce sullo spettatore. Lungo tale percorso, Bettetini affronta un nodo cruciale degli studi attuali sulla comunicazione audiovisiva, offrendo allo stesso tempo un prezioso compendio della letteratura semiotica sull’argomento. Nella prima parte del volume lo studioso definisce le linee essenziali di un nuovo approccio metodologico che integra, con la sociologia e la psicologia, il quadro delle discipline che si sono occupate sistematicamente di analisi del testo filmico dal secondo dopoguerra in avanti. Nella seconda parte l’autore prende in esame quattro lungometraggi: Caterina va in città (2003 di Paolo Virzì, Notturno Bus (2007 di Davide Marengo, Il flauto magico (2007 di Kenneth Branagh e La fine del mondo – Ministoria della musica in un tempo (1969 dello stesso Bettetini.

  9. Time-dependent, bidirectional, anti- and pro-spinal hyper-reflexia and muscle spasticity effect after chronic spinal glycine transporter 2 (GlyT2) oligonucleotide-induced downregulation.

    Kamizato, Kota; Marsala, Silvia; Navarro, Michael; Kakinohana, Manabu; Platoshyn, Oleksandr; Yoshizumi, Tetsuya; Lukacova, Nadezda; Wancewicz, Ed; Powers, Berit; Mazur, Curt; Marsala, Martin

    2018-07-01

    The loss of local spinal glycine-ergic tone has been postulated as one of the mechanisms contributing to the development of spinal injury-induced spasticity. In our present study using a model of spinal transection-induced muscle spasticity, we characterize the effect of spinally-targeted GlyT2 downregulation once initiated at chronic stages after induction of spasticity in rats. In animals with identified hyper-reflexia, the anti-spasticity effect was studied after intrathecal treatment with: i) glycine, ii) GlyT2 inhibitor (ALX 1393), and iii) GlyT2 antisense oligonucleotide (GlyT2-ASO). Administration of glycine and GlyT2 inhibitor led to significant suppression of spasticity lasting for a minimum of 45-60 min. Treatment with GlyT2-ASO led to progressive suppression of muscle spasticity seen at 2-3 weeks after treatment. Over the subsequent 4-12 weeks, however, the gradual appearance of profound spinal hyper-reflexia was seen. This was presented as spontaneous or slight-tactile stimulus-evoked muscle oscillations in the hind limbs (but not in upper limbs) with individual hyper-reflexive episodes lasting between 3 and 5 min. Chronic hyper-reflexia induced by GlyT2-ASO treatment was effectively blocked by intrathecal glycine. Immunofluorescence staining and Q-PCR analysis of the lumbar spinal cord region showed a significant (>90%) decrease in GlyT2 mRNA and GlyT2 protein. These data demonstrate that spinal GlyT2 downregulation provides only a time-limited therapeutic benefit and that subsequent loss of glycine vesicular synthesis resulting from chronic GlyT2 downregulation near completely eliminates the tonic glycine-ergic activity and is functionally expressed as profound spinal hyper-reflexia. These characteristics also suggest that chronic spinal GlyT2 silencing may be associated with pro-nociceptive activity. Copyright © 2018 Elsevier Inc. All rights reserved.

  10. Data on the purification and crystallization of the loss-of-function von Willebrand disease variant (p.Gly1324Ser of the von Willebrand factor A1 domain

    James C. Cambell

    2016-06-01

    In this data article we describe the production, quality control and crystallization of the p.Gly1324Ser vWD variant of the A1 domain of VWF. p.Gly1324Ser A1 was expressed in Escherichia coli as insoluble inclusion bodies. After the preparation of the inclusion bodies, the protein was solubilized, refolded, purified by affinity chromatography and crystallized. The crystal structure of the p.Gly1324Ser mutant of the A1 domain is deposited at the Protein Data Bank PDB: 5BV8

  11. Factor VII Tokushima: the first case of factor VII Cys22Gly with the development of myocardial infarction in the proband receiving recombinant factor VIIa replacement therapy.

    Shigekiyo, Toshio; Sekimoto, Etsuko; Shibata, Hironobu; Ozaki, Shuji; Okumura, Takanobu; Fujinaga, Hiroyuki; Shibata, Hiroshi; Aihara, Ken-ichi; Akaike, Masashi

    2015-12-01

    An 81-year-old man was referred to our department because of suspected factor VII (FVII) deficiency. His FVII activity was under 1%, whereas the FVII activity levels of his son and granddaughter were 65 and 109%, respectively. The nucleotide at position 3886 of his FVII gene was homozygous for G. A single T to G substitution results in the replacement of wild-type Cys at residue 22 by Gly. His son was heterozygous for G and T at position 3886, whereas his granddaughter was homozygous for wild-type T. These results suggest that he was homozygous for FVII Cys22Gly. He underwent radiofrequency ablation (RFA) for hepatocellular carcinoma, receiving 20 μg/kg of recombinant FVIIa prior to RFA and 10 μg/kg of recombinant FVIIa twice after RFA. He showed no bleeding tendency; however, a myocardial infarction was diagnosed and percutaneous coronary intervention was performed.

  12. Mutation of Gly717Phe in human topoisomerase 1B has an effect on enzymatic function, reactivity to the camptothecin anticancer drug and on the linker domain orientation

    Wang, Zhenxing; D'Annessa, Ilda; Tesauro, Cinzia

    2015-01-01

    –DNA covalent adduct. In this work the role of the Gly717 residue, located in a α-helix structure bridging the active site and the linker domain, has been investigated mutating it in Phe. The mutation gives rise to drug resistance in vivo as observed through a viability assay of yeast cells. In vitro activity...... assays show that the mutant is characterized by a fast religation rate, only partially reduced by the presence of the drug. Comparative molecular dynamics simulations of the native and mutant proteins indicate that the mutation of Gly717 affects the motion orientation of the linker domain, changing its...... interaction with the DNA substrate, likely affecting the strand rotation and religation rate. The mutation also causes a slight rearrangement of the active site and of the drug binding site, providing an additional explanation for the lowered effect of camptothecin toward the mutant....

  13. Anticancer drugs and the regulation of Hedgehog genes GLI1 and PTCH1, a comparative study in nonmelanoma skin cancer cell lines

    Olesen, Uffe H; Bojesen, Sophie; Gehl, Julie

    2017-01-01

    Nonmelanoma skin cancer is the most common cancer in humans, comprising mainly basal cell carcinoma (BCC) and squamous cell carcinoma (SCC). BCC proliferation is highly dependent on the Hedgehog signaling pathway. We aimed to investigate a panel of anticancer drugs with known activity against skin...... of immortalized keratinocytes (HaCaT), BCC (UWBCC1 and BCC77015), and SCC (A431 and SCC25) cell lines. The impact of treatment on the regulation of Hedgehog pathway target genes (GLI1 and PTCH1), measured by real-time PCR, was compared between UWBCC1 and HaCaT. Varying cell line sensitivity profiles...... to the examined anticancer drugs were observed. Generally, 24-h drug exposure was sufficient to reduce cell viability. We found that 5-FU, MTX, and cisplatin significantly downregulated the expression of two genes controlled by the Hedgehog pathway (≤25-, 2.9-, and 12.5-fold, respectively, for GLI1 in UWBCC1...

  14. Ask the users, il valore aggiunto della valutazione dei sistemi informativi culturali on line coinvolgendo gli utenti: il caso del progetto “Una Città per gli Archivi” / Ask the users, the added value of the evaluation of on-line cultural information systems involving users: the case of “Una città per gli archvi” project

    Pierluigi Feliciati

    2012-10-01

    Digital environments aimed at offering services on historical records and documents put together archival domain standards with the typical features of digital libraries, with item-level descriptions, access to digital objects, subject-based searching. How can on line archives move from the plain delivery of structured information to the facilitation of a cultural experience? This communication presents some of these issues, with a focus on the peculiarities of web displays for archival descriptions and reproductions. In this field, the role played by user studies is crucial, to guarantee that return on investment whose first profit should be users satisfaction. After a short review of the international state of the art the author will present an on-going user study project, launched to test the prototype of Una Città per gli Archivi, the web portal disseminating the results of a huge project of preservation and valorization of documentary funds of the city of Bologna, including photographic and graphic materials, sound and audiovisual recordings, in addition to paper archives.

  15. A Gly1127Ser mutation in an EGF-like domain of the Fibrillin-I gene is a risk factor for ascending aortic aneurysm and dissection

    Francke, U.; Berg, M.A.; Tynan, K. [Stanford Univ. Medical Center, CA (United States)] [and others

    1995-06-01

    Ascending aortic disease, ranging from mild aortic root enlargement to aneurysm and/or dissection, has been identified in 10 individuals of a kindred, none of whom had classical Marfan syndrome (MFS). Single-strand conformation analysis of the entire fibrillin-1 (FBN1) cDNA of an affected family member revealed a G-to-A transition at nucleotide 3379, predicting a Gly1127Ser substitution. The glycine in this position is highly conserved in EGF-like domains of FBN1 and other proteins. This mutation was present in 9 of 10 affected family members and in 1 young unaffected member but was not found in other unaffected members, in 168 chromsomes from normal controls, and in 188 chromosomes from other individuals with MFS or related phenotypes. FBN1 intragenic marker haplotypes ruled out the possibility that the other allele played a significant role in modulating the phenotype in this family. Pulse-chase studies revealed normal fibrillin synthesis but reduced fibrillin deposition into the extracellular matrix in cultured fibroblasts from a Gly1127Ser carrier. We postulate that the Gly1127Ser FBN1 mutation is responsible for reduced matrix deposition. We suggest that mutations such as this one may disrupt EFG-like domain folding less drastically than do substitutions of cysteine or of other amino acids important for calcium-binding that cause classical MFS. The Gly 1127Ser mutation, therefore, produces a mild form of autosomal dominantly inherited weakness of elastic tissue, which predisposes to ascending aortic aneurysm and dissection later in life. 33 refs., 6 figs.

  16. Transmembrane potential of GlyCl-expressing instructor cells induces a neoplastic-like conversion of melanocytes via a serotonergic pathway

    Douglas Blackiston

    2011-01-01

    Understanding the mechanisms that coordinate stem cell behavior within the host is a high priority for developmental biology, regenerative medicine and oncology. Endogenous ion currents and voltage gradients function alongside biochemical cues during pattern formation and tumor suppression, but it is not known whether bioelectrical signals are involved in the control of stem cell progeny in vivo. We studied Xenopus laevis neural crest, an embryonic stem cell population that gives rise to many cell types, including melanocytes, and contributes to the morphogenesis of the face, heart and other complex structures. To investigate how depolarization of transmembrane potential of cells in the neural crest’s environment influences its function in vivo, we manipulated the activity of the native glycine receptor chloride channel (GlyCl. Molecular-genetic depolarization of a sparse, widely distributed set of GlyCl-expressing cells non-cell-autonomously induces a neoplastic-like phenotype in melanocytes: they overproliferate, acquire an arborized cell shape and migrate inappropriately, colonizing numerous tissues in a metalloprotease-dependent fashion. A similar effect was observed in human melanocytes in culture. Depolarization of GlyCl-expressing cells induces these drastic changes in melanocyte behavior via a serotonin-transporter-dependent increase of extracellular serotonin (5-HT. These data reveal GlyCl as a molecular marker of a sparse and heretofore unknown cell population with the ability to specifically instruct neural crest derivatives, suggest transmembrane potential as a tractable signaling modality by which somatic cells can control stem cell behavior at considerable distance, identify a new biophysical aspect of the environment that confers a neoplastic-like phenotype upon stem cell progeny, reveal a pre-neural role for serotonin and its transporter, and suggest a novel strategy for manipulating stem cell behavior.

  17. Flexible xxx-asp/asn and gly-xxx residues of equine cytochrome C in matrix-assisted laser desorption/ionization in-source decay mass spectrometry.

    Takayama, Mitsuo

    2012-01-01

    The backbone flexibility of a protein has been studied from the standpoint of the susceptibility of amino acid residues to in-source decay (ISD) in matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). Residues more susceptible to MALDI-ISD, namely Xxx-Asp/Asn and Gly-Xxx, were identified from the discontinuous intense peak of c'-ions originating from specific cleavage at N-Cα bonds of the backbone of equine cytochrome c. The identity of the residues susceptible to ISD was consistent with the known flexible backbone amides as estimated by hydrogen/deuterium exchange (HDX) experiments. The identity of these flexible amino acid residues (Asp, Asn, and Gly) is consistent with the fact that these residues are preferred in flexible secondary structure free from intramolecular hydrogen-bonded structures such as α-helix and β-sheet. The MALDI-ISD spectrum of equine cytochrome c gave not only intense N-terminal side c'-ions originating from N-Cα bond cleavage at Xxx-Asp/Asn and Gly-Xxx residues, but also C-terminal side complement z'-ions originating from the same cleavage sites. The present study implies that MALDI-ISD can give information about backbone flexibility of proteins, comparable with the protection factors estimated by HDX.

  18. Retrieval of precipitable water using near infrared channels of Global Imager/Advanced Earth Observing Satellite-II (GLI/ADEOS-II)

    Kuji, M.; Uchiyama, A.

    2002-01-01

    Retrieval of precipitable water (vertically integrated water vapor amount) is proposed using near infrared channels og Global Imager onboard Advanced Earth Observing Satellite-II (GLI/ADEOS-II). The principle of retrieval algorithm is based upon that adopted with Moderate Resolution Imaging Spectroradiometer (MODIS) onboard Earth Observing System (EOS) satellite series. Simulations were carried out with GLI Signal Simulator (GSS) to calculate the radiance ratio between water vapor absorbing bands and non-absorbing bands. As a result, it is found that for the case of high spectral reflectance background (a bright target) such as the land surface, the calibration curves are sensitive to the precipitable water variation. For the case of low albedo background (a dark target) such as the ocean surface, on the contrary, the calibration curve is not very sensitive to its variation under conditions of the large water vapor amount. It turns out that aerosol loading has little influence on the retrieval over a bright target for the aerosol optical thickness less than about 1.0 at 500nm. It is also anticipated that simultaneous retrieval of the water vapor amount using GLI data along with other channels will lead to improved accuracy of the determination of surface geophysical properties, such as vegetation, ocean color, and snow and ice, through the better atmospheric correction

  19. Dye adsorbates BrPDI, BrGly, and BrAsp on anatase TiO2(001) for dye-sensitized solar cell applications

    Çakır, D.; Gülseren, O.; Mete, E.; Ellialtıoǧlu, Ş.

    2009-07-01

    Using the first-principles plane-wave pseudopotential method within density functional theory, we systematically investigated the interaction of perylenediimide (PDI)-based dye compounds (BrPDI, BrGly, and BrAsp) with both unreconstructed (UR) and reconstructed (RC) anatase TiO2(001) surfaces. All dye molecules form strong chemical bonds with surface in the most favorable adsorption structures. In UR-BrGly, RC-BrGly, and RC-BrAsp cases, we have observed that highest occupied molecular orbital and lowest unoccupied molecular orbital levels of molecules appear within band gap and conduction-band region, respectively. Moreover, we have obtained a gap narrowing upon adsorption of BrPDI on the RC surface. Because of the reduction in effective band gap of surface-dye system and possibly achieving the visible-light activity, these results are valuable for photovoltaic and photocatalytic applications. We have also considered the effects of hydration of surface to the binding of BrPDI. It has been found that the binding energy drops significantly for the completely hydrated surfaces.

  20. Alpha-adducin Gly460Trp polymorphism and hypertension risk: a meta-analysis of 22 studies including 14303 cases and 15961 controls.

    Kuo Liu

    Full Text Available BACKGROUND: No clear consensus has been reached on the alpha-adducin polymorphism (Gly460Trp and essential hypertension risk. We performed a meta-analysis in an effort to systematically summarize the possible association. METHODOLOGY/PRINCIPAL FINDINGS: Studies were identified by searching MEDLINE and EMBASE databases complemented with perusal of bibliographies of retrieved articles and correspondence with original authors. The fixed-effects model and the random-effects model were applied for dichotomous outcomes to combine the results of the individual studies. We selected 22 studies that met the inclusion criteria including a total of 14303 hypertensive patients and 15961 normotensive controls. Overall, the 460Trp allele showed no statistically significant association with hypertension risk compared to Gly460 allele (P = 0.69, OR = 1.02, 95% CI 0.94-1.10, P(heterogeneity<0.0001 in all subjects. Meta-analysis under other genetic contrasts still did not reveal any significant association in all subjects, Caucasians, East Asians and others. The results were similar but heterogeneity did not persist when sensitivity analyses were limited to these studies. CONCLUSIONS/SIGNIFICANCE: Our meta-analysis failed to provide evidence for the genetic association of α-adducin gene Gly460Trp polymorphism with hypertension. Further studies investigating the effect of genetic networks, environmental factors, individual biological characteristics and their mutual interactions are needed to elucidate the possible mechanism for hypertension in humans.

  1. Toll receptor 4 Asp299Gly polymorphism and its association with preterm birth and premature rupture of membranes in a South American population.

    Rey, G; Skowronek, F; Alciaturi, J; Alonso, J; Bertoni, B; Sapiro, R

    2008-09-01

    Preterm birth (PTB) is a worldwide health problem and remains the leading cause of perinatal morbidity and mortality. Systemic and local intrauterine infections have been implicated in the pathogenesis of preterm labor and delivery. Common pathways between PTB, premature rupture of ovular membranes (PROM) and altered molecular routes of inflammation have been proposed. There is evidence to support a genetic component in these conditions. Lipopolysaccharide (LPS), a component of the cell wall of Gram-negative bacteria, is thought to play a key role in eliciting an inflammatory response. LPS is recognized by proteins of the innate immune system, including Toll-like receptor 4 (TLR4). Individuals from some European countries carrying the variant alleles resulting in an amino acid substitution (Asp299Gly) are at increased risk of Gram-negative infections and premature birth. The objective of this study was to determine if preterm newborns have different allele frequency of the Asp299Gly TLR4 variant from healthy term neonates in Uruguay. The impact of PROM was also examined. There was an increase in the risk for fetuses carrying the Asp299Gly substitution in TLR4 of being severely premature (<33 weeks) and to present PROM at the same time.

  2. Evaluation of a novel Arg-Gly-Asp-conjugated α-melanocyte stimulating hormone hybrid peptide for potential melanoma therapy.

    Yang, Jianquan; Guo, Haixun; Gallazzi, Fabio; Berwick, Marianne; Padilla, R Steven; Miao, Yubin

    2009-08-19

    The purpose of this study was to determine whether Arg-Gly-Asp (RGD)-conjugated α-melanocyte stimulating hormone (α-MSH) hybrid peptide could be employed to target melanocortin-1 (MC1) receptor for potential melanoma therapy. The RGD motif {cyclic(Arg-Gly-Asp-DTyr-Asp)} was coupled to [Cys(3,4,10), DPhe(7), Arg(11)]α-MSH(3-13) {(Arg(11))CCMSH} to generate RGD-Lys-(Arg(11))CCMSH hybrid peptide. The MC1 receptor binding affinity of RGD-Lys-(Arg(11))CCMSH was determined in B16/F1 melanoma cells. The internalization and efflux, melanoma targeting and pharmacokinetic properties and single photon emission computed tomography/CT (SPECT/CT) imaging of (99m)Tc-RGD-Lys-(Arg(11))CCMSH were determined in B16/F1 melanoma cells and melanoma-bearing C57 mice. Clonogenic cytotoxic effect of RGD-Lys-(Arg(11))CCMSH was examined in B16/F1 melanoma cells. RGD-Lys-(Arg(11))CCMSH displayed 2.1 nM MC1 receptor binding affinity. (99m)Tc-RGD-Lys-(Arg(11))CCMSH showed rapid internalization and extended retention in B16/F1 cells. The cellular uptake of (99m)Tc-RGD-Lys-(Arg(11))CCMSH was MC1 receptor-mediated. (99m)Tc-RGD-Lys-(Arg(11))CCMSH exhibited high tumor uptake (14.83 ± 2.94% ID/g 2 h postinjection) and prolonged tumor retention (7.59 ± 2.04% ID/g 24 h postinjection) in B16/F1 melanoma-bearing mice. Nontarget organ uptakes were generally low except for the kidneys. Whole-body clearance of (99m)Tc-RGD-Lys-(Arg(11))CCMSH was rapid, with approximately 62% of the injected radioactivity cleared through the urinary system by 2 h postinjection. Flank melanoma tumors were clearly imaged by small animal SPECT/CT using (99m)Tc-RGD-Lys-(Arg(11))CCMSH as an imaging probe 2 h postinjection. Single treatment (3 h incubation) with 100 nM of RGD-Lys-(Arg(11))CCMSH significantly (p < 0.05) decreased the clonogenic survival of B16/F1 cells by 65% compared to the untreated control cells. Favorable melanoma targeting property of (99m)Tc-RGD-Lys-(Arg(11))CCMSH and remarkable cytotoxic effect of RGD

  3. Gli affari di messer Palla Strozzi (e di suo padre Nofri. Imprenditoria e mecenatismo nella Firenze del primo Rinascimento

    Sergio Tognetti

    2011-10-01

    Full Text Available Il saggio si propone di far luce sulle vicende economico-patrimoniali di messer Palla di Nofri Strozzi (Firenze 1372-Padova 1462, personaggio di spicco della Firenze del primo Rinascimento, sia come uomo politico ingiustamente travolto dalla purga medicea scatenata nel 1434 contro gli oppositori di Cosimo il Vecchio, sia come letterato, collezionista di codici librari, promotore degli studia humanitatis e committente di opere d’arte. Al centro di una frammentata serie di indagini da parte di storici dell’arte, della cultura e della politica, Palla Strozzi ha suscitato invece scarsa attenzione da parte degli storici dell’economia, nonostante un particolare non secondario: al catasto fiorentino del 1427 risultava essere il cittadino più ricco, ma dopo nemmeno cinque anni era già così oberato dai debiti da non riuscire più a pagare le tasse al fisco fiorentino. La vicenda di messer Palla, messa a confronto con quella del padre Nofri, infaticabile uomo d’affari capace di ammassare una gigantesca fortuna (poi parzialmente dissipata dal figlio, è analizzata nei dettagli grazie alla ricchissima serie archivistica delle carte strozziane: in particolare tramite lo spoglio di numerosi libri di conti intestati ad aziende e patrimoni di famiglia. Il confronto tra le differenti attitudini dei due personaggi fa emergere in tutta la sua originalità la figura di messer Palla: uomo coltissimo ma anche elitario, eppure dotato di un alto senso dello stato, capace di mostrare verso il denaro un’inclinazione che lo avvicina più all’aristocratico rentier dell’ancien régime e/o al raffinato signore padano del pieno rinascimento, piuttosto che al mondo dei grandi «mercatanti» che lo aveva generato.

  4. Immunoproteomics based identification of thioredoxin reductase GliT and novel Aspergillus fumigatus antigens for serologic diagnosis of invasive aspergillosis.

    Shi, Li-ning; Li, Fang-qiu; Huang, Mei; Lu, Jing-fen; Kong, Xiao-xiang; Wang, Shi-qin; Shao, Hai-feng

    2012-01-18

    There has been a rising incidence of invasive aspergillosis (IA) in critically ill patients, even in the absence of an apparent predisposing immunodeficiency. The diagnosis of IA is difficult because clinical signs are not sensitive and specific, and serum galactomannan has relatively low sensitivity in this group of patients. Therefore, more prompt and accurate disease markers for early diagnosis are needed. To establish disease markers demands a thorough knowledge of fungal antigens which may be detected in the serum or other body fluids of patients. Herein we report novel immunodominant antigens identified from extracellular proteins of Aspergillus fumigatus. Extracellular proteins of A. fumigatus were separated by two-dimensional electrophoresis (2-DE) and probed with the sera from critically ill patients with proven IA. The immunoreactive protein spots were identified by MALDI-TOF mass spectrometry (MALDI-TOF -MS). Forty spots from 2DE gels were detected and 17 different proteins were identified as immunogenic in humans. Function annotation revealed that most of these proteins were metabolic enzymes involved in carbohydrate, fatty acid, amino acid, and energy metabolism. One of the proteins, thioredoxin reductase GliT (TR), which showed the best immunoactivity, was analyzed further for secretory signals, protein localization, and homology. The results indicated that TR is a secretory protein with a signal sequence exhibiting a high probability for secretion. Furthermore, TR did not match any human proteins, and had low homology with most other fungi. The recombinant TR was recognized by the sera of all proven IA patients with different underlying diseases in this study. The immunoreactive proteins identified in this study may be helpful for the diagnosis of IA in critically ill patients. Our results indicate that TR and other immunodominant antigens have potential as biomarkers for the serologic diagnosis of invasive aspergillosis.

  5. Immunoproteomics based identification of thioredoxin reductase GliT and novel Aspergillus fumigatus antigens for serologic diagnosis of invasive aspergillosis

    Shi Li-ning

    2012-01-01

    Full Text Available Abstract Background There has been a rising incidence of invasive aspergillosis (IA in critically ill patients, even in the absence of an apparent predisposing immunodeficiency. The diagnosis of IA is difficult because clinical signs are not sensitive and specific, and serum galactomannan has relatively low sensitivity in this group of patients. Therefore, more prompt and accurate disease markers for early diagnosis are needed. To establish disease markers demands a thorough knowledge of fungal antigens which may be detected in the serum or other body fluids of patients. Herein we report novel immunodominant antigens identified from extracellular proteins of Aspergillus fumigatus. Results Extracellular proteins of A. fumigatus were separated by two-dimensional electrophoresis (2-DE and probed with the sera from critically ill patients with proven IA. The immunoreactive protein spots were identified by MALDI-TOF mass spectrometry (MALDI-TOF -MS. Forty spots from 2DE gels were detected and 17 different proteins were identified as immunogenic in humans. Function annotation revealed that most of these proteins were metabolic enzymes involved in carbohydrate, fatty acid, amino acid, and energy metabolism. One of the proteins, thioredoxin reductase GliT (TR, which showed the best immunoactivity, was analyzed further for secretory signals, protein localization, and homology. The results indicated that TR is a secretory protein with a signal sequence exhibiting a high probability for secretion. Furthermore, TR did not match any human proteins, and had low homology with most other fungi. The recombinant TR was recognized by the sera of all proven IA patients with different underlying diseases in this study. Conclusions The immunoreactive proteins identified in this study may be helpful for the diagnosis of IA in critically ill patients. Our results indicate that TR and other immunodominant antigens have potential as biomarkers for the serologic diagnosis

  6. Gly[14]-humanin inhibits ox-LDL uptake and stimulates cholesterol efflux in macrophage-derived foam cells.

    Zhu, Wa-Wa; Wang, Shu-Rong; Liu, Zhi-Hua; Cao, Yong-Jun; Wang, Fen; Wang, Jing; Liu, Chun-Feng; Xie, Ying; Xie, Ying; Zhang, Yan-Lin

    2017-01-01

    Foam cell formation, which is caused by imbalanced cholesterol influx and efflux by macrophages, plays a vital role in the occurrence and development of atherosclerosis. Humanin (HN), a mitochondria-derived peptide, can prevent the production of reactive oxygen species and death of human aortic endothelial cells exposed to oxidized low-density lipoprotein (ox-LDL) and has a protective effect on patients with in early atherosclerosis. However, the effects of HN on the regulation of cholesterol metabolism in RAW 264.7 macrophages are still unknown. This study was designed to investigate the role of [Gly14]-humanin (HNG) in lipid uptake and cholesterol efflux in RAW 264.7 macrophages. Flow cytometry and live cell imaging results showed that HNG reduced Dil-ox-LDL accumulation in the RAW 264.7 macrophages. A similar result was obtained for lipid accumulation by measuring cellular cholesterol content. Western blot analysis showed that ox-LDL treatment upregulated not only the protein expression of CD36 and LOX-1, which mediate ox-LDL endocytosis, but also ATP-binding cassette (ABC) transporter A1 and ABCG1, which mediate ox-LDL exflux. HNG pretreatment inhibited the upregulation of CD36 and LOX-1 levels, prompting the upregulation of ABCA1 and ABCG1 levels induced by ox-LDL. Therefore we concluded that HNG could inhibit ox-LDL-induced macrophage-derived foam cell formation, which occurs because of a decrease in lipid uptake and an increase in cholesterol efflux from macrophage cells. Copyright © 2016 Elsevier Inc. All rights reserved.

  7. I big data e gli strumenti di visualizzazione analitica: interazioni e studi induttivi per le P.A.

    Giuseppe Roccasalva

    2012-12-01

    Full Text Available Il saggio presenta alcuni risultati di una collaborazione tra Politecnico di Torino e il CSI Piemonte (Società di servizi Informatizzati partecipata dalla Regione Piemonte. Sono stati selezionati e studiati diversi strumenti di visualizzazione dei dati scientifici (Gapminder, ManyEyes, Open eXplorer e Fineo al fine di individuare quello più utile per una lettura induttiva di grandi quantità di dati informativi (big data. Lo sfruttamento intelligente dei dati digitali può portare a uno sviluppo conoscitivo ma anche a un profitto, le cui soglie di sfruttamento possono essere misurate in un sistema economico. Nell’irreversibile fenomeno di crescita dei dati digitali, la disciplina del “Data Visualization” diventa cruciale per accedere e comprendere informazioni complesse. Few, guru della comunicazione visiva, scrive che “scopriamo il mondo attraverso gli occhi”; le forme di comunicazione e interpretazione tradizionali dei dati hanno puntato sulla dimensione visuale per migliorare la comprensione e hanno permesso sia agli analisti sia agli utenti la sperimentazione di nuove interazioni (“story-telling”. Come urbanisti e cittadini, ci affidiamo alla vista che gestisce molti dei sensori (70% legati alla percezione, alle mappe cognitive, agli errori e ai nuovi pensieri. L’ipotesi di fondo di questo articolo vuole generare delle riflessioni sui Big Data come strategia importante per le imprese pubbliche e private che intendono imparare a cambiare dalle informazioni digitali di cui oggi disponiamo. Attraverso l’uso di uno strumento analitico di visualizzazione dei dati informativi, si descrive un recente caso di studio in un contesto territoriale come quello dei nuovi consorzi amministrativi (Unione dei Comuni NordEst Torino. In questo esperimento torna a essere attuale la necessità di pianificare le scelte in modo sistematico anche cercando di utilizzare in modo nuovo e semplice i sistemi informativi territoriali già disponibili.

  8. The Rho ADP-ribosylating C3 exoenzyme binds cells via an Arg-Gly-Asp motif.

    Rohrbeck, Astrid; Höltje, Markus; Adolf, Andrej; Oms, Elisabeth; Hagemann, Sandra; Ahnert-Hilger, Gudrun; Just, Ingo

    2017-10-27

    The Rho ADP-ribosylating C3 exoenzyme (C3bot) is a bacterial protein toxin devoid of a cell-binding or -translocation domain. Nevertheless, C3 can efficiently enter intact cells, including neurons, but the mechanism of C3 binding and uptake is not yet understood. Previously, we identified the intermediate filament vimentin as an extracellular membranous interaction partner of C3. However, uptake of C3 into cells still occurs (although reduced) in the absence of vimentin, indicating involvement of an additional host cell receptor. C3 harbors an Arg-Gly-Asp (RGD) motif, which is the major integrin-binding site, present in a variety of integrin ligands. To check whether the RGD motif of C3 is involved in binding to cells, we performed a competition assay with C3 and RGD peptide or with a monoclonal antibody binding to β1-integrin subunit and binding assays in different cell lines, primary neurons, and synaptosomes with C3-RGD mutants. Here, we report that preincubation of cells with the GRGDNP peptide strongly reduced C3 binding to cells. Moreover, mutation of the RGD motif reduced C3 binding to intact cells and also to recombinant vimentin. Anti-integrin antibodies also lowered the C3 binding to cells. Our results indicate that the RGD motif of C3 is at least one essential C3 motif for binding to host cells and that integrin is an additional receptor for C3 besides vimentin. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  9. The sequential action of a dipeptidase and a beta-lyase is required for the release of the human body odorant 3-methyl-3-sulfanylhexan-1-ol from a secreted Cys-Gly-(S) conjugate by Corynebacteria.

    Emter, Roger; Natsch, Andreas

    2008-07-25

    Human axillary odor is formed by the action of Corynebacteria on odorless axilla secretions. Sulfanylalkanols, 3-methyl-3-sulfanylhexan-1-ol in particular, form one key class of the odoriferous compounds. A conjugate with the dipeptide Cys-Gly has been reported as the secreted precursor for 3-methyl-3-sulfanylhexan-1-ol. Here, we confirm the Cys-Gly-(S) conjugate as the major precursor of this odorant, with lower levels of the Cys-(S) conjugate being present in axilla secretions. The enzymatic release of 3-methyl-3-sulfanylhexan-1-ol from the Cys-Gly-(S) conjugate by the axilla isolate Corynebacterium Ax20 was thus investigated. Cellular extracts of Ax20 released 3-methyl-3-sulfanylhexan-1-ol from the Cys-Gly-(S) conjugate and from the Cys-(S) conjugate, whereas the previously isolated C-S lyase of this bacterial strain was only able to cleave the Cys-(S) conjugate. o-Phenanthroline blocked the release from the Cys-Gly-(S) conjugate but did not affect cleavage of the Cys-(S) conjugate, indicating that in a first step, a metal-dependent dipeptidase hydrolyzes the Cys-Gly bond. This enzyme was purified by four chromatographic steps and gel electrophoresis, and the partial amino acid sequence was determined. The corresponding gene was cloned and expressed in Escherichia coli. It codes for a novel dipeptidase with a high affinity toward the Cys-Gly-(S) conjugate of 3-methyl-3-sulfanylhexan-1-ol. Co-incubating either the synthetic Cys-Gly-(S) conjugate or fresh axilla secretions with both the C-S lyase and the novel dipeptidase did release 3-methyl-3-sulfanylhexan-1-ol, proving that the sequential action of these two enzymes from the skin bacterium Corynebacterium Ax20 does release the odorant from the key secreted precursor.

  10. Gli Archetipi della Fantasia

    Michela De Domenico

    2015-12-01

    Full Text Available In comics, visions of fantastic architecture contributed, along visions of utopia architecture in movies and video games, to the creation of an imaginary fantastic architecture. At the base of these creations there’s an ideational process starting from drawings and that allows to see things and locations not existing yet. In such – not fully explored – context, target is to investigate themes related to ephemeral and fantasy then set up tools for analysis through the research of main archetypes, the identification of the grammar and semiotic who communise message of such architectures. Target is to show, by the mutual exchages among cities and comics, the existence of a cross area inspired to a memory of common architecture, to a similar imaginary, and showing a common interest for ambient mutations.

  11. Gly184 of the Escherichia coli cAMP receptor protein provides optimal context for both DNA binding and RNA polymerase interaction.

    Hicks, Matt N; Gunasekara, Sanjiva; Serate, Jose; Park, Jin; Mosharaf, Pegah; Zhou, Yue; Lee, Jin-Won; Youn, Hwan

    2017-10-01

    The Escherichia coli cAMP receptor protein (CRP) utilizes the helix-turn-helix motif for DNA binding. The CRP's recognition helix, termed F-helix, includes a stretch of six amino acids (Arg180, Glu181, Thr182, Val183, Gly184, and Arg185) for direct DNA contacts. Arg180, Glu181 and Arg185 are known as important residues for DNA binding and specificity, but little has been studied for the other residues. Here we show that Gly184 is another F-helix residue critical for the transcriptional activation function of CRP. First, glycine was repeatedly selected at CRP position 184 for its unique ability to provide wild type-level transcriptional activation activity. To dissect the glycine requirement, wild type CRP and mutants G184A, G184F, G184S, and G184Y were purified and their in vitro DNA-binding activity was measured. G184A and G184F displayed reduced DNA binding, which may explain their low transcriptional activation activity. However, G184S and G184Y displayed apparently normal DNA affinity. Therefore, an additional factor is needed to account for the diminished transcriptional activation function in G184S and G184Y, and the best explanation is perturbations in their interaction with RNA polymerase. The fact that glycine is the smallest amino acid could not fully warrant its suitability, as shown in this study. We hypothesize that Gly184 fulfills the dual functions of DNA binding and RNA polymerase interaction by conferring conformational flexibility to the F-helix.

  12. Progressive Encephalomyelitis with Rigidity and Myoclonus Associated With Anti-GlyR Antibodies and Hodgkin’s Lymphoma: A Case Report

    Linda Borellini

    2017-08-01

    Full Text Available IntroductionA 60-year-old man presented with a 6-month history of low back pain and progressive rigidity of the trunk and lower limbs, followed by pruritus, dysphonia, hyperhydrosis, and urinary retention. Brain and spinal imaging were normal. EMG showed involuntary motor unit hyperactivity. Onconeural, antiglutamic acid decarboxylase (anti-GAD, voltage-gated potassium channel, and dipeptidyl peptidase-like protein 6 (DPPX autoantibodies were negative. CSF was negative. Symptoms were partially responsive to baclofen, gabapentin, and clonazepam, but he eventually developed severe dysphagia. Antiglycine receptor (anti-GlyR antibodies turned out positive on both serum and CSF. A plasmapheresis cycle was completed with good clinical response. A PET scan highlighted an isolated metabolically active axillary lymphnode that turned out to be a classic type Hodgkin lymphoma (HL, in the absence of bone marrow infiltration nor B symptoms. Polychemotherapy with ABVD protocol was completed with good clinical response and at 1-year follow-up the neurological examination is normal.BackgroundProgressive encephalomyelitis with rigidity and myoclonus (PERM is a rare and severe neurological syndrome characterized by muscular rigidity and spasms as well as brain stem and autonomic dysfunction. It can be associated with anti-GAD, GlyR, and DPPX antibodies. All of these autoantibodies may be variably associated with malignant tumors and their response to immunotherapy, as well as to tumor removal, is not easily predictable.ConclusionProgressive encephalomyelitis with rigidity and myoclonus has already been described in association with HL, but this is the first case report of a HL manifesting as anti-GlyR antibodies related PERM. Our report highlights the importance of malignancy screening in autoimmune syndromes of suspected paraneoplastic origin.

  13. The new industrial political tools based on the sustainable development of productive activities; Gli strumenti attuativi di una politica di sviluppo sostenibile dell'impresa

    Luciani, R.; Andriola, L. [ENEA, Centro Ricerche Casaccia, Rome (Italy). Dipt. Ambiente

    1999-07-01

    The new industrial political tools based on the sustainable development of productive activities, and originating at the Rio Conference in 1992, late introduced by the European Union, the Government and voluntary organizations (ISO, CEM, UNI) are gradually entering the market, thereby underling their effect in terms of industrial competitiveness. The main tools were examined in the report and mostly refer to the voluntary ones which, in surpassing the command and control logic, increasingly tend to get close to traditional fiscal, tariff and financial ones. Voluntary agreements, environmental management systems (EMAS, ISO14000), the risk prevention systems (UNI 10617), certification (Ecolabel, ISO 14040) and environment communications tools (environment balance sheet, environmental report, environmental statement) are synthetically described. Finally, the tools regarding the incentives still under Government concern an their diffusion are examined. [Italian] I nuovi strumenti di politica industriale orientati allo sviluppo sostenibile delle attivita' produttive che traggono origine dalla Conferenza di Rio del 1992 e sono stati introdotti successivamente da parte della Comunita' Europea, dal Governo e dagli organismi di normazione volontaria (ISO, CEN, UNI), stanno gradualmente penetrando sul mercato facendo valere i loro effetti anche in termini di competitivita' delle imprese. Nel rapporto sono stati esaminati i principali tra questi strumenti con particolare riferimento a quelli volontari che sempre piu', nel superamento della logica del {sup c}ommand-control{sup ,} tendono ad affiancarsi a quelli tradizionali di carattere fiscale, tariffario e finanziario. In particolare vengono descritti sinteticamente gli accordi volontari, i sistemi di gestione ambientali e della sicurezza (EMAS, norme ISO 14001 e UNI 10617), la certificazione di prodotto (Ecolabel, norma ISO 14040) e gli strumenti di comunicazione ambientale (bilancio ambientale, rapporto

  14. Encapsulation of rat bone marrow stromal cells using a poly-ion complex gel of chitosan and succinylated poly(Pro-Hyp-Gly).

    Kusumastuti, Yuni; Shibasaki, Yoshiaki; Hirohara, Shiho; Kobayashi, Mime; Terada, Kayo; Ando, Tsuyoshi; Tanihara, Masao

    2017-03-01

    Encapsulation of stem cells into a three-dimensional (3D) scaffold is necessary to achieve tissue regeneration. Prefabricated 3D scaffolds, such as fibres or porous sponges, have limitations regarding homogeneous cell distribution. Hydrogels that can encapsulate cells such as animal-derived collagen gels need adjustment of the pH and/or temperature upon cell mixing. In this report, we fabricated a poly-ion complex (PIC) hydrogel of chitosan and succinylated poly(Pro-Hyp-Gly) and assessed its effect on cell viability after encapsulation of rat bone marrow stromal cells. PIC hydrogels were obtained successfully with a concentration of each precursor as low as 3.0-3.8 mg/ml. The maximum gelation and swelling ratios were achieved with an equal molar ratio (1:1) of anionic and cationic groups. Using chitosan acetate as a cationic precursor produced a PIC hydrogel with both a significantly greater gelation ratio and a better swelling ratio than chitosan chloride. Ammonium succinylated poly(Pro-Hyp-Gly) as an anionic precursor gave similar gelation and swelling ratios to those of sodium succinylated poly(Pro-Hyp-Gly). Cell encapsulation was also achieved successfully by mixing rat bone marrow stromal cells with the PIC hydrogel simultaneously during its formation. The PIC hydrogel was maintained in the culture medium for 7 days at 37°C and the encapsulated cells survived and proliferated in it. Although it is necessary to improve its functionality, this PIC hydrogel has the potential to act as a 3D scaffold for cell encapsulation and tissue regeneration. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

  15. The new industrial political tools based on the sustainable development of productive activities; Gli strumenti attuativi di una politica di sviluppo sostenibile dell'impresa

    Luciani, R; Andriola, L [ENEA, Centro Ricerche Casaccia, Rome (Italy). Dipt. Ambiente

    1999-07-01

    The new industrial political tools based on the sustainable development of productive activities, and originating at the Rio Conference in 1992, late introduced by the European Union, the Government and voluntary organizations (ISO, CEM, UNI) are gradually entering the market, thereby underling their effect in terms of industrial competitiveness. The main tools were examined in the report and mostly refer to the voluntary ones which, in surpassing the command and control logic, increasingly tend to get close to traditional fiscal, tariff and financial ones. Voluntary agreements, environmental management systems (EMAS, ISO14000), the risk prevention systems (UNI 10617), certification (Ecolabel, ISO 14040) and environment communications tools (environment balance sheet, environmental report, environmental statement) are synthetically described. Finally, the tools regarding the incentives still under Government concern an their diffusion are examined. [Italian] I nuovi strumenti di politica industriale orientati allo sviluppo sostenibile delle attivita' produttive che traggono origine dalla Conferenza di Rio del 1992 e sono stati introdotti successivamente da parte della Comunita' Europea, dal Governo e dagli organismi di normazione volontaria (ISO, CEN, UNI), stanno gradualmente penetrando sul mercato facendo valere i loro effetti anche in termini di competitivita' delle imprese. Nel rapporto sono stati esaminati i principali tra questi strumenti con particolare riferimento a quelli volontari che sempre piu', nel superamento della logica del {sup c}ommand-control{sup ,} tendono ad affiancarsi a quelli tradizionali di carattere fiscale, tariffario e finanziario. In particolare vengono descritti sinteticamente gli accordi volontari, i sistemi di gestione ambientali e della sicurezza (EMAS, norme ISO 14001 e UNI 10617), la certificazione di prodotto (Ecolabel, norma ISO 14040) e gli strumenti di comunicazione ambientale (bilancio ambientale, rapporto ambientale

  16. Altering adsorbed proteins or cellular gene expression in bone-metastatic cancer cells affects PTHrP and Gli2 without altering cell growth

    Jonathan M. Page

    2015-09-01

    Full Text Available The contents of this data in brief are related to the article titled “Matrix Rigidity Regulates the Transition of Tumor Cells to a Bone-Destructive Phenotype through Integrin β3 and TGF-β Receptor Type II”. In this DIB we will present our supplemental data investigating Integrin expression, attachment of cells to various adhesion molecules, and changes in gene expression in multiple cancer cell lines. Since the interactions of Integrins with adsorbed matrix proteins are thought to affect the ability of cancer cells to interact with their underlying substrates, we examined the expression of Integrin β1, β3, and β5 in response to matrix rigidity. We found that only Iβ3 increased with increasing substrate modulus. While it was shown that fibronectin greatly affects the expression of tumor-produced factors associated with bone destruction (parathyroid hormone-related protein, PTHrP, and Gli2, poly-l-lysine, vitronectin and type I collagen were also analyzed as potential matrix proteins. Each of the proteins was independently adsorbed on both rigid and compliant polyurethane films which were subsequently used to culture cancer cells. Poly-l-lysine, vitronectin and type I collagen all had negligible effects on PTHrP or Gli2 expression, but fibronectin was shown to have a dose dependent effect. Finally, altering the expression of Iβ3 demonstrated that it is required for tumor cells to respond to the rigidity of the matrix, but does not affect other cell growth or viability. Together these data support the data presented in our manuscript to show that the rigidity of bone drives Integrinβ3/TGF-β crosstalk, leading to increased expression of Gli2 and PTHrP.

  17. Structure-activity relationship of linear peptide Bu-His-DPhe-Arg-Trp-Gly-NH(2) at the human melanocortin-1 and -4 receptors: arginine substitution.

    Cheung, Adrian Wai-Hing; Danho, Waleed; Swistok, Joseph; Qi, Lida; Kurylko, Grazyna; Franco, Lucia; Yagaloff, Keith; Chen, Li

    2002-09-02

    A series of pentapeptides, based on Bu-His(6)-DPhe(7)-Arg(8)-Trp(9)-Gly(10)-NH(2) and modified at the Arg(8) position, was prepared and pharmacologically characterized. Peptides containing either cyanoguanidine or acylguanidine, two substantially less basic arginine surrogates, were found to retain the agonist activity of the parent peptide at both hMC1R and hMC4R. This study unequivocally shows that the positive charge of Arg(8) is not essential for efficient interactions of our pentapeptide with both hMC1R and hMC4R.

  18. Potent μ-Opioid Receptor Agonists from Cyclic Peptides Tyr-c[D-Lys-Xxx-Tyr-Gly]: Synthesis, Biological, and Structural Evaluation.

    Li, Yangmei; Cazares, Margret; Wu, Jinhua; Houghten, Richard A; Toll, Laurence; Dooley, Colette

    2016-02-11

    To optimize the structure of a μ-opioid receptor ligand, analogs H-Tyr-c[D-Lys-Xxx-Tyr-Gly] were synthesized and their biological activity was tested. The analog containing a Phe(3) was identified as not only exhibiting binding affinity 14-fold higher than the original hit but also producing agonist activity 3-fold more potent than morphine. NMR study suggested that a trans conformation at D-Lys(2)-Xxx(3) is crucial for these cyclic peptides to maintain high affinity, selectivity, and functional activity toward the μ-opioid receptor.

  19. Comparative proteomic analyses reveal that FlbA down-regulates gliT expression and SOD activity in Aspergillus fumigatus.

    Shin, Kwang-Soo; Park, Hee-Soo; Kim, Young-Hwan; Yu, Jae-Hyuk

    2013-07-11

    FlbA is a regulator of G-protein signaling protein that plays a central role in attenuating heterotrimeric G-protein mediated vegetative growth signaling in Aspergillus. The deletion of flbA (∆flbA) in the opportunistic human pathogen Aspergillus fumigatus results in accelerated cell death and autolysis in submerged culture. To further investigate the effects of ∆flbA on intracellular protein levels we carried out 2-D proteome analyses of 2-day old submerged cultures of ∆flbA and wild type (WT) strains and observed 160 differentially expressed proteins. Via nano-LC-ESI-MS/MS analyses, we revealed the identity of 10 and 2 proteins exhibiting high and low level accumulation, respectively, in ∆flbA strain. Notably, the GliT protein is accumulated at about 1800-fold higher levels in ∆flbA than WT. Moreover, GliT is secreted at high levels from ∆flbA strain, whereas Sod1 (superoxide dismutase) is secreted at a higher level in WT. Northern blot analyses reveal that ∆flbA results in elevated accumulation of gliT mRNA. Consequently, ∆flbA strain exhibits enhanced tolerance to gliotoxin toxicity. Finally, ∆flbA strain displayed enhanced SOD activity and elevated resistance to menadione and paraquat. In summary, FlbA-mediated signaling control negatively affects cellular responses associated with detoxification of reactive oxygen species and of exogenous gliotoxin in A. fumigatus. Regulator of G protein Signaling (RGS) proteins play crucial roles in fundamental biological processes in filamentous fungi. FlbA is the first studied filamentous fungal RGS protein, yet much remains to be understood about its roles in the opportunistic human pathogen Aspergillus fumigatus. In the present study, we examined the effects of the deletion of flbA using comprehensive analyses of the intra- and extracellular proteomes of A. fumigatus wild type and the flbA deletion mutant. Via MS analyses, we identified 10 proteins exhibiting high level accumulation in the flbA deletion

  20. Cell adhesion to fibrillin-1: identification of an Arg-Gly-Asp-dependent synergy region and a heparin-binding site that regulates focal adhesion formation

    Bax, Daniel V; Mahalingam, Yashithra; Cain, Stuart

    2007-01-01

    We have defined the molecular basis of cell adhesion to fibrillin-1, the major structural component of extracellular microfibrils that are associated with elastic fibres. Using human dermal fibroblasts, and recombinant domain swap fragments containing the Arg-Gly-Asp motif, we have demonstrated...... a requirement for upstream domains for integrin-alpha(5)beta(1)-mediated cell adhesion and migration. An adjacent heparin-binding site, which supports focal adhesion formation, was mapped to the fibrillin-1 TB5 motif. Site-directed mutagenesis revealed two arginine residues that are crucial for heparin binding...

  1. A novel germ-line point mutation in RET exon 8 (Gly(533)Cys) in a large kindred with familial medullary thyroid carcinoma

    Silva, Adriana Madeira Alvares da [UNIFESP; Maciel, Rui Monteiro de Barros [UNIFESP; Dias-da-Silva, Magnus Régios [UNIFESP; Toledo, Silvia Regina Caminada de [UNIFESP; De Carvalho, Marcos B.; Cerutti, Janete Maria [UNIFESP

    2003-01-01

    Familial medullary thyroid carcinoma is related to germ-line mutations in the RET oncogene, mainly in cysteine codon 10 or 11, whereas noncysteine mutations in codons 13 - 15 are rare. We now report a new missense point mutation in exon 8 of the RET gene (1597G-->T) corresponding to a Gly(533)Cys substitution in the cystein-rich domain of RET protein in 76 patients from a 6-generation Brazilian family with 229 subjects, with ascendants from Spain. It is likely that the mutation causes familia...

  2. Improving Tumor Uptake and Pharmacokinetics of 64Cu-Labeled Cyclic RGD Peptide Dimers with Gly3 and PEG4 Linkers

    Shi, Jiyun; Kim, Young-Seung; Zhai, Shizhen; Liu, Zhaofei; Chen, Xiaoyuan; Liu, Shuang

    2009-01-01

    Radiolabeled cyclic RGD (Arg-Gly-Asp) peptides represent a new class of radiotracers with potential for the early tumor detection and non-invasive monitoring of tumor metastasis and therapeutic response in cancer patients. This report describes the synthesis of two cyclic RGD peptide dimer conjugates, DOTA-PEG4-E[PEG4-c(RGDfK)]2 (DOTA-3PEG4-dimer: DOTA = 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid; PEG4 = 15-amino-4,7,10,13-tetraoxapentadecanoic acid) and DOTA-G3-E[G3-c(RGDfK)]2 ...

  3. Role of ASCA and the NOD2/CARD15 mutation Gly908Arg in predicting increased surgical costs in Crohn's disease patients: a project of the European Collaborative Study Group on Inflammatory Bowel Disease.

    Odes, Shmuel; Friger, Michael; Vardi, Hillel; Claessens, Greet; Bossuyt, Xavier; Riis, Lene; Munkholm, Pia; Wolters, Frank; Yona, Hagit; Hoie, Ole; Beltrami, Marina; Tsianos, Epameinondas; Katsanos, Kostas; Mouzas, Ioannis; Clofent, Juan; Monteiro, Estela; Messori, Andrea; Politi, Patrizia; O'Morain, Colm; Limonard, Charles; Russel, Maurice; Vatn, Morten; Moum, Bjorn; Stockbrugger, Reinhold; Vermeire, Severine

    2007-07-01

    NOD2/CARD15, the first identified susceptibility gene in Crohn's disease (CD), is associated with ileal stenosis and increased frequency of surgery. Anti-Saccharomyces cerevisiae antibody (ASCA), a serological marker for CD, is associated with ileal location and a high likelihood for surgery. We hypothesized that the presence of ASCA and NOD2/CARD15 mutations could predict increased health care cost in CD. CD patients in a prospectively designed community-based multinational European and Israeli cohort (n = 228) followed for mean 8.3 (SD 2.6) years had blood drawn for measurement of ASCA (IgG, IgA), Arg702Trp, Gly908Arg, and Leu1007fsinsC. Days spent in the hospital and the costs of medical and surgical hospitalizations and medications were calculated. The median duration of surgical hospitalizations was longer in Gly908Arg-positive than -negative patients, 3.5 and 1.5 days/patient-year (P < 0.01), and in ASCA-positive than -negative patients, 1.1 and 0 days/patient-year (P < 0.001). Median surgical hospitalization cost was 1,580 euro/patient-year in Gly908Arg-positive versus 0 euro/patient-year in -negative patients (P < 0.01), and 663 euro/patient-year in ASCA-positive versus 0 euro/patient-year in -negative patients (P < 0.001). Differences in cost of medications between groups were not significant. The effect of Gly908Arg was expressed in countries with higher Gly908Arg carriage rates. ASCA raised surgical costs independently of the age at diagnosis of disease. Arg702Trp and Leu1007fsinsC did not affect the cost of health care. Since CD patients positive for Gly908Arg and ASCA demonstrated higher health care costs, it is possible that measurement of Gly908Arg and ASCA at disease diagnosis can forecast the expensive CD patients.

  4. Tritium labelling of highly selective probes for. delta. -opioid receptors: ( sup 3 H)Tyr-D-Ser(O-t-Bu)-Gly-Phe-Leu-Thr(DSTBULET) and ( sup 3 H)Tyr-D-Ser(O-t-Bu)-Gly-Phe-Leu-Thr(O-t-Bu)(BUBU)

    Fellion, E.; Gacel, G.; Roques, B.P. (Institut National de la Sante et de la Recherche Medicale (INSERM), 75 - Paris (France)); Roy, J.; Morgat, J.L. (CEA Centre d' Etudes Nucleaires de Saclay, 91 - Gif-sur-Yvette (France). Service de Biochimie)

    1990-08-01

    The introduction of bulky residue(s) in linear enkephalin-related hexapeptides represents a new approach in the design of selective probes for {delta}-opioid receptors, displaying the appropriate criteria to investigate biological and pharmacological properties of the assumed binding site ({delta}) of endogenous enkephalins. The selectivities and high affinities of Tyr-D-Ser(O-t-Bu)-Gly-Phe-Leu-Thr(DSTBULET) and especially Tyr-D-Ser(O-t-Bu)Gly-Phe-Leu-Thr(O-t-Bu) (BUBU) associated with a satisfactory resistance to peptidases, make them the most suitable {delta}-probes reported to date. In the present paper, we report the synthesis of DSTBULET and BUBU under tritiated forms with high specific radioactivities. These radio-labelled probes will enable extensive in vitro and in vivo investigations of {delta}-opioid receptors properties to be carried out. (author).

  5. Thermo-chemotherapy Induced miR-218 upregulation inhibits the invasion of gastric cancer via targeting Gli2 and E-cadherin.

    Ruan, Qiang; Fang, Zhi-Yuan; Cui, Shu-Zhong; Zhang, Xiang-Liang; Wu, Yin-Bing; Tang, Hong-Sheng; Tu, Yi-Nuo; Ding, Yan

    2015-08-01

    Thermo-chemotherapy has been proven to reduce the invasion capability of cancer cells. However, the molecular mechanism underlying this anti-invasion effect is still unclear. In this study, the role of thermo-chemotherapy in the inhibition of tumor invasion was studied. The results demonstrated that expression of miR-218 was downregulated in gastric cancer tissues, which had a positive correlation with tumor invasion and metastasis. In vitro thermo-chemotherapy increased miR-218 expression in SGC7901 cells and inhibited both proliferation and invasion of cancer cells. Gli2 was identified as a downstream target of miR-218, and its expression was negatively regulated by miR-218. The thermo-chemotherapy induced miR-218 upregulation was also accompanied by increasing of E-cadherin expression. In conclusion, the present study indicates that thermo-chemotherapy can effectively decrease the invasion capability of cancer cells and increase cell-cell adhesion. miR-218 and its downstream target Gli2, as well as E-cadherin, participate in the anti-invasion process.

  6. The single nucleotide polymorphism Gly482Ser in the PGC-1α gene impairs exercise-induced slow-twitch muscle fibre transformation in humans.

    Peter Steinbacher

    Full Text Available PGC-1α (peroxisome proliferator-activated receptor γ co-activator 1α is an important regulator of mitochondrial biogenesis and a master regulator of enzymes involved in oxidative phosphorylation. Recent evidence demonstrated that the Gly482Ser single nucleotide polymorphism (SNP in the PGC-1α gene affects insulin sensitivity, blood lipid metabolism and binding to myocyte enhancer factor 2 (MEF2. Individuals carrying this SNP were shown to have a reduced cardiorespiratory fitness and a higher risk to develop type 2 diabetes. Here, we investigated the responses of untrained men with the Gly482Ser SNP to a 10 week programme of endurance training (cycling, 3 x 60 min/week, heart rate at 70-90% VO2peak. Quantitative data from analysis of biopsies from vastus lateralis muscle revealed that the SNP group, in contrast to the control group, lacked a training-induced increase in content of slow contracting oxidative fibres. Capillary supply, mitochondrial density, mitochondrial enzyme activities and intramyocellular lipid content increased similarly in both groups. These results indicate that the impaired binding of MEF2 to PGC-1α in humans with this SNP impedes exercise-induced fast-to-slow muscle fibre transformation.

  7. The single nucleotide polymorphism Gly482Ser in the PGC-1α gene impairs exercise-induced slow-twitch muscle fibre transformation in humans.

    Steinbacher, Peter; Feichtinger, René G; Kedenko, Lyudmyla; Kedenko, Igor; Reinhardt, Sandra; Schönauer, Anna-Lena; Leitner, Isabella; Sänger, Alexandra M; Stoiber, Walter; Kofler, Barbara; Förster, Holger; Paulweber, Bernhard; Ring-Dimitriou, Susanne

    2015-01-01

    PGC-1α (peroxisome proliferator-activated receptor γ co-activator 1α) is an important regulator of mitochondrial biogenesis and a master regulator of enzymes involved in oxidative phosphorylation. Recent evidence demonstrated that the Gly482Ser single nucleotide polymorphism (SNP) in the PGC-1α gene affects insulin sensitivity, blood lipid metabolism and binding to myocyte enhancer factor 2 (MEF2). Individuals carrying this SNP were shown to have a reduced cardiorespiratory fitness and a higher risk to develop type 2 diabetes. Here, we investigated the responses of untrained men with the Gly482Ser SNP to a 10 week programme of endurance training (cycling, 3 x 60 min/week, heart rate at 70-90% VO2peak). Quantitative data from analysis of biopsies from vastus lateralis muscle revealed that the SNP group, in contrast to the control group, lacked a training-induced increase in content of slow contracting oxidative fibres. Capillary supply, mitochondrial density, mitochondrial enzyme activities and intramyocellular lipid content increased similarly in both groups. These results indicate that the impaired binding of MEF2 to PGC-1α in humans with this SNP impedes exercise-induced fast-to-slow muscle fibre transformation.

  8. Investigation of the spatial structure of des-Gly9-[Arg8]vasopressin by the methods of two-dimensional NMR spectroscopy and theoretical conformational analysis

    Shenderovich, M.D.; Sekatsis, I.P.; Liepin'sh, E.E.; Nikiforovich, G.V.; Papsuevich, O.S.

    1986-01-01

    An assignment of the 1 H NMR signals of des-Gly 9 -[Arg 8 ]vasopressin in dimethyl sulfoxide has been made by 2D spectroscopy. The SSCCs and temperature coefficients Δδ/Δ T have been obtained for the amide protons and the system of NOE cross-peaks in the two-dimensional NOESY spectrum has been analyzed. The most important information on the spatial structure of des-Gly 9 -[Arg 8 ]vasopressin is given by the low value of the temperature coefficient Δδ/Δ T of the Asn 5 amide proton and the NOE between the α-protons of Cys 1 and Cys 6 . It is assumed that the screening of the NH proton of the Asn 5 residue from the solvent is connected with a β-bend of the backbone in the 2-5 sequence, and the distance between the C/sup α/H atoms of the Cys 1 and Cys 6 residues does not exceed 4 A. Bearing these limitations in mind, a theoretical conformational analysis of the molecule has been made. The group of low-energy conformations of the backbone obtained has been compared with the complete set of NMR characteristics

  9. Gli1-Mediated Regulation of Sox2 Facilitates Self-Renewal of Stem-Like Cells and Confers Resistance to EGFR Inhibitors in Non-Small Cell Lung Cancer.

    Bora-Singhal, Namrata; Perumal, Deepak; Nguyen, Jonathan; Chellappan, Srikumar

    2015-07-01

    Non-small cell lung cancer (NSCLC) patients have very low survival rates because the current therapeutic strategies are not fully effective. Although EGFR tyrosine kinase inhibitors are effective for NSCLC patients harboring EGFR mutations, patients invariably develop resistance to these agents. Alterations in multiple signaling cascades have been associated with the development of resistance to EGFR inhibitors. Sonic Hedgehog and associated Gli transcription factors play a major role in embryonic development and have recently been found to be reactivated in NSCLC, and elevated Gli1 levels correlate with poor prognosis. The Hedgehog pathway has been implicated in the functions of cancer stem cells, although the underlying molecular mechanisms are not clear. In this context, we demonstrate that Gli1 is a strong regulator of embryonic stem cell transcription factor Sox2. Depletion of Gli1 or inhibition of the Hedgehog signaling significantly abrogated the self-renewal of stem-like side-population cells from NSCLCs as well as vascular mimicry of such cells. Gli1 was found to transcriptionally regulate Sox2 through its promoter region, and Gli1 could be detected on the Sox2 promoter. Inhibition of Hedgehog signaling appeared to work cooperatively with EGFR inhibitors in markedly reducing the viability of NSCLC cells as well as the self-renewal of stem-like cells. Thus, our study demonstrates a cooperative functioning of the EGFR signaling and Hedgehog pathways in governing the stem-like functions of NSCLC cancer stem cells and presents a novel therapeutic strategy to combat NSCLC harboring EGFR mutations. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  10. Regulation of sonic hedgehog-GLI1 downstream target genes PTCH1, Cyclin D2, Plakoglobin, PAX6 and NKX2.2 and their epigenetic status in medulloblastoma and astrocytoma

    Eberhart Charles G

    2010-11-01

    Full Text Available Abstract Background The Sonic hedgehog (Shh signaling pathway is critical for cell growth and differentiation. Impairment of this pathway can result in both birth defects and cancer. Despite its importance in cancer development, the Shh pathway has not been thoroughly investigated in tumorigenesis of brain tumors. In this study, we sought to understand the regulatory roles of GLI1, the immediate downstream activator of the Shh signaling pathway on its downstream target genes PTCH1, Cyclin D2, Plakoglobin, NKX2.2 and PAX6 in medulloblastoma and astrocytic tumors. Methods We silenced GLI1 expression in medulloblastoma and astrocytic cell lines by transfection of siRNA against GLI1. Subsequently, we performed RT-PCR and quantitative real time RT-PCR (qRT-PCR to assay the expression of downstream target genes PTCH1, Cyclin D2, Plakoglobin, NKX2.2 and PAX6. We also attempted to correlate the pattern of expression of GLI1 and its regulated genes in 14 cell lines and 41 primary medulloblastoma and astrocytoma tumor samples. We also assessed the methylation status of the Cyclin D2 and PTCH1 promoters in these 14 cell lines and 58 primary tumor samples. Results Silencing expression of GLI1 resulted up-regulation of all target genes in the medulloblastoma cell line, while only PTCH1 was up-regulated in astrocytoma. We also observed methylation of the cyclin D2 promoter in a significant number of astrocytoma cell lines (63% and primary astrocytoma tumor samples (32%, but not at all in any medulloblastoma samples. PTCH1 promoter methylation was less frequently observed than Cyclin D2 promoter methylation in astrocytomas, and not at all in medulloblastomas. Conclusions Our results demonstrate different regulatory mechanisms of Shh-GLI1 signaling. These differences vary according to the downstream target gene affected, the origin of the tissue, as well as epigenetic regulation of some of these genes.

  11. Regulation of sonic hedgehog-GLI1 downstream target genes PTCH1, Cyclin D2, Plakoglobin, PAX6 and NKX2.2 and their epigenetic status in medulloblastoma and astrocytoma

    Shahi, Mehdi H; Afzal, Mohammad; Sinha, Subrata; Eberhart, Charles G; Rey, Juan A; Fan, Xing; Castresana, Javier S

    2010-01-01

    The Sonic hedgehog (Shh) signaling pathway is critical for cell growth and differentiation. Impairment of this pathway can result in both birth defects and cancer. Despite its importance in cancer development, the Shh pathway has not been thoroughly investigated in tumorigenesis of brain tumors. In this study, we sought to understand the regulatory roles of GLI1, the immediate downstream activator of the Shh signaling pathway on its downstream target genes PTCH1, Cyclin D2, Plakoglobin, NKX2.2 and PAX6 in medulloblastoma and astrocytic tumors. We silenced GLI1 expression in medulloblastoma and astrocytic cell lines by transfection of siRNA against GLI1. Subsequently, we performed RT-PCR and quantitative real time RT-PCR (qRT-PCR) to assay the expression of downstream target genes PTCH1, Cyclin D2, Plakoglobin, NKX2.2 and PAX6. We also attempted to correlate the pattern of expression of GLI1 and its regulated genes in 14 cell lines and 41 primary medulloblastoma and astrocytoma tumor samples. We also assessed the methylation status of the Cyclin D2 and PTCH1 promoters in these 14 cell lines and 58 primary tumor samples. Silencing expression of GLI1 resulted up-regulation of all target genes in the medulloblastoma cell line, while only PTCH1 was up-regulated in astrocytoma. We also observed methylation of the cyclin D2 promoter in a significant number of astrocytoma cell lines (63%) and primary astrocytoma tumor samples (32%), but not at all in any medulloblastoma samples. PTCH1 promoter methylation was less frequently observed than Cyclin D2 promoter methylation in astrocytomas, and not at all in medulloblastomas. Our results demonstrate different regulatory mechanisms of Shh-GLI1 signaling. These differences vary according to the downstream target gene affected, the origin of the tissue, as well as epigenetic regulation of some of these genes

  12. Positive selection at codon 38 of the human KCNE1 (= minK gene and sporadic absence of 38Ser-coding mRNAs in Gly38Ser heterozygotes

    Pfeufer Arne

    2009-08-01

    Full Text Available Abstract Background KCNE1 represents the regulatory beta-subunit of the slowly activating delayed rectifier potassium channel (IKs. Variants of KCNE1 have repeatedly been linked to the long-QT syndrome (LQTS, a disorder which predisposes to deafness, ventricular tachyarrhythmia, syncope, and sudden cardiac death. Results We here analyze the evolution of the common Gly38Ser variant (rs1805127, using genomic DNAs, complementary DNAs, and HEK293-expressed variants of altogether 19 mammalian species. The between species comparison reveals that the human-specific Gly38Ser polymorphism evolved under strong positive Darwinian selection, probably in adaptation to specific challenges in the fine-tuning of IKs channels. The involved amino acid exchanges (Asp > Gly, Gly > Ser are moderately radical and do not induce apparent changes in posttranslational modification. According to population genetic analyses (HapMap phase II a heterozygote advantage accounts for the maintenance of the Gly38Ser polymorphism in humans. On the other hand, the expression of the 38Ser allele seems to be disadvantageous under certain conditions, as suggested by the sporadic deficiency of 38Ser-coding mRNAs in heterozygote Central Europeans and the depletion of homozygotes 38Ser in the Yoruban sample. Conclusion We speculate that individual differences in genomic imprinting or genomic recoding might have contributed to conflicting results of recent association studies between Gly38Ser polymorphism and QT phenotype. The findings thus highlight the relevance of mRNA data in future association studies of genotypes and clinical disorders. To the best of our knowledge, they moreover provide first time evidence for a unique pattern; i.e. coincidence of positive Darwinian selection and polymorphism with a sporadically suppressed expression of one allele.

  13. Targeting a Cross-Reactive Gly m 5 Soy Peptide as Responsible for Hypersensitivity Reactions in a Milk Allergy Mouse Model

    Curciarello, Renata; Smaldini, Paola L.; Candreva, Angela M.; González, Virginia; Parisi, Gustavo; Cauerhff, Ana; Barrios, Ivana; Blanch, Luis Bruno; Fossati, Carlos A.

    2014-01-01

    Background Cross-reactivity between soybean allergens and bovine caseins has been previously reported. In this study we aimed to map epitopes of the major soybean allergen Gly m 5 that are co-recognized by casein specific antibodies, and to identify a peptide responsible for the cross-reactivity. Methods Cow's milk protein (CMP)-specific antibodies were used in different immunoassays (immunoblotting, ELISA, ELISA inhibition test) to evaluate the in vitro recognition of soybean proteins (SP). Recombinant Gly m 5 (α), a truncated fragment containing the C-terminal domain (α-T) and peptides of α-T were obtained and epitope mapping was performed with an overlapping peptide assay. Bioinformatics tools were used for epitope prediction by sequence alignment, and for modelling the cross-recognized soy proteins and peptides. The binding of SP to a monoclonal antibody was studied by surface Plasmon resonance (SPR). Finally, the in vivo cross-recognition of SP was assessed in a mouse model of milk allergy. Results Both α and α-T reacted with the different CMP-specific antibodies. α-T contains IgG and IgE epitopes in several peptides, particularly in the peptide named PA. Besides, we found similar values of association and dissociation constants between the α-casein specific mAb and the different milk and soy components. The food allergy mouse model showed that SP and PA contain the cross-reactive B and T epitopes, which triggered hypersensitivity reactions and a Th2-mediated response on CMP-sensitized mice. Conclusions Gly m 5 is a cross-reactive soy allergen and the α-T portion of the molecule contains IgG and IgE immunodominant epitopes, confined to PA, a region with enough conformation to be bound by antibodies. These findings contribute to explain the intolerance to SP observed in IgE-mediated CMA patients, primarily not sensitised to SP, as well as it sets the basis to propose a mucosal immunotherapy for milk allergy using this soy peptide. PMID:24416141

  14. Variantes polimórficas Ala513Pro y Gly972Arg del gen IRS-1 no se asocian a la diabetes mellitus tipo 2 en un grupo de la población cubana The Ala513Pro and Gly972ARg polymorphous variants of IRS-1 gen are not associated with type diabetes mellitus in a group of the Cuban population

    Luis Miguel Pérez

    2011-08-01

    Full Text Available Introducción: la diabetes mellitus tipo 2 es una enfermedad heterogénea y multifactorial, que está determinada por factores genéticos y no genéticos. El sustrato 1 del receptor de la insulina (IRS-1 cumple una función fundamental en la transmisión de la señal insulínica, por tanto sus variantes génicas constituyen blancos importantes en el estudio de la susceptibilidad genética a esta enfermedad en las diferentes poblaciones. Objetivo: explorar el papel de las variantes polimórficas Gly972Arg y Ala513Pro del gen IRS-1 en la susceptibilidad genética de la diabetes mellitus tipo 2 en un grupo de la población cubana. Métodos: se determinó la frecuencia de los polimorfismos Gly972Arg y Ala513Pro del IRS-1 en 499 ciudadanos cubanos, con un índice de masa corporal entre 22-30, con edades comprendidas entre los 40 y 70 años: de ellos 272 (54,5 % diabéticos y 227 (45,5 % no diabéticos. Resultados: la frecuencia del alelo Pro513 fue baja (1,2 % y similar para ambos grupos (1,1 % vs. 1,3 % para el grupo de diabéticos y el grupo control, respectivamente. La frecuencia del polimorfismo Gly972Arg fue de 16,2%, superior a la reportada para la mayoría de las poblaciones estudiadas. No se encontraron diferencias significativas en la frecuencia del alelo Arg972 entre el grupo de diabéticos y el grupo control (15,4 % vs. 17,3 %, ni cambios en los niveles de glucemia e insulinemia asociados a la presencia del alelo polimórfico Arg972. Conclusiones: en este grupo de sujetos de la población cubana, las variantes polimórficas Ala513Pro y Gly972Arg del gen IRS-1 no participan en la etiología de la diabetes mellitus tipo 2.Introduction: the type 2 diabetes mellitus is a heterogeneous and multifactor disease determined by genetic and no-genetic factors. The substrate 1 of insulin receptor (IRS-1 has a fundamental function in transmission of insulin signal, thus its genic variants are significant targets in study of genetic susceptibility to

  15. The increased incidence of the RET p.Gly691Ser variant in French-Canadian vesicoureteric reflux patients is not replicated by a larger study in Ireland.

    Darlow, John M

    2012-02-01

    The p.Gly691Ser variant of the RET protein, resulting from the \\'A\\' allele of the SNP rs1799939 in exon 11 of the RET gene, was recently found to be present in a high proportion of primary vesicoureteric reflux (pVUR) patients in Quebec. We have determined the genotype of this SNP in 221 unrelated index cases of pVUR from the Irish population, in 190 full siblings of 160 of the index cases, and in 592 healthy controls. We found no significant difference in genotype or allele frequencies in patients and controls, and no tendency of affected siblings to share the same genotype. We also found no difference in the presence of additional phenotypic features such as duplex kidneys, between patients with and without the \\'A\\' allele, and no difference in grade of reflux. We find no evidence of any influence of RET SNP rs1799939 on pVUR phenotype.

  16. Osteogenesis imperfecta Type IV: a newly identified variant at position c.560 (G > T; p.Gly187Val) in the COL1A2 gene.

    Usta, Akin; Karademir, Dilay; Sen, Eylem; Yazici, Selcuk; Adali, Ertan; Erdem, Erkan; Karacan, Meric

    2017-01-01

    Osteogenesis imperfecta is a clinically heterogenous disease caused by defective collagen syntesis associated with a mutation in the COL1A1 or COL1A2 genes. In this report, we present a case of osteogenesis imperfecta (OI) type IV, seen in a female fetus with incurved femurs at 18 weeks of gestation. Molecular analysis of the newborn revealed a novel mutation at position c.560 (c.560 G > T) of the exon 12 in the COL1A2 gene; which lead to the glycine modification with valine (p.Gly187Val) at codon 187. The pregnancy follow-up was uneventful. After delivery, the newborn underwent biphosponat therapy and no fracture was detected until 1 year old.

  17. In-depth evaluation of Gly-Sar transport parameters as a function of culture time in the Caco-2 cell model

    Bravo, Silvina A.; Nielsen, Carsten Uhd; Amstrup, Jan

    2004-01-01

    The aim of the present study was to investigate the influence of culture time on hPEPT1-mediated transport in Caco-2 cell monolayers. Peptide transport activity in Caco-2 cells grown in standard media and in a "rapid" 4-day model was first compared. The rapid 4-day Caco-2 cell model, cultured using...... a cocktail of growth factors and agonists, displayed lower peptide uptake capacity than Caco-2 cells grown for 4 days in conventional media, and was judged to be unsuitable for peptide transport studies. Peptide transport activity as well as monolayer integrity and tissue morphology were evaluated...... in the standard >21 days model as a function of the culture time. Peptide transport activity was studied using [14C]-glycylsarcosine ([ 14C]-Gly-Sar). Monolayer integrity was evaluated by transepithelial electrical resistance (TEER) measurements and [3H]-mannitol permeabilities. Tissue morphology and hPEPT1...

  18. Synthesis of the tritium labelled β-casomorphine analogues 3H-Phe-Pro-Gly-OH and 3H2-Tyr-Pro-3H-Phe-pyrrolidide

    Oehlke, J.; Niedrich, H.; Born, I.; Neubert, K.; Mittag, E.

    1991-01-01

    The precursor peptides H-Phe(I)-Pro-Gly-OH (III) and H-Tyr(I 2 )-Pro-Phe(I)-pyrrolidide (VIII) were synthesized by stepwise elongation from the C-terminal end and by coupling of Boc-Tyr(I 2 )-Pro-OH with H-Phe(I)-pyrrolidide and following deprotection of the Boc-residue respectively. Catalytic dehalotritiation yielded tritated peptides with specific radioactivities of 450 and 1500 GBq/mmol respectively. Cleavage of 3 H 2 -Tyr-Pro- 3 H-Phe-pyrrolidide by dipeptidylpeptidase IV resulted in fragments with specific radioactivities of 950 ( 3 H 2 -Tyr-Pro) and 590 GBq/ mmol ( 3 H-Phe-pyrrolidide). (author)

  19. Environmental analysis applied to schools. Methodologies for data acquisition; Analisi ambientale iniziale per gli istituti d'istruzione. Metodologia per l'acquisizione dei dati

    Andriola, L.; Ceccacci, R. [ENEA, Div. Caratterizzazione dell' Ambiente e del Territorio, Centro Ricerche Casaccia, Rome (Italy)

    2001-07-01

    The environment analysis is the basis of environmental management for organizations and it is considered as the first step in EMAS. It allows to identify, deal with the issues and have a clear knowledge on environmental performances of organizations. Schools can be included in the organizations. Nevertheless, the complexity of environmental issues and applicable regulations makes very difficult for a school, that wants to implement an environmental management system (EMAS, ISO 14001, etc.), to face this first step. So, it has been defined an instrument, that is easy but complete and coherent with reference standard, to let schools choose their process for elaborating the initial environmental revue. This instrument consists, essentially, in cards that, if completed, facilitate the drafting of the environmental analysis report. [Italian] L'analisi ambientale iniziale costituisce il presupposto di qualsiasi azione di gestione ambientale da parte di un'organizzazione ed e' esplicitamente prevista, come prima tappa, in EMAS. Essa consente infatti di individuare ed approfondire le problematiche ambientali e di fotografare, per mezzo di riferimenti oggettivi, le prestazioni ambientali di un'organizzazione, e quindi anche di una scuola, in modo da poter stabilire e controllare nel tempo gli obiettivi di miglioramento. Tuttavia la complessita' delle problematiche ambientali e delle relative normative di riferimento rende estremamente difficoltoso, per un istituto scolastico che intenda intraprendere un percorso di gestione ambientale comunque finalizzato (EMAS, ISO 14001, Comunicazione ambientale, ecc.) compiere questo passo iniziale. E' stato pertanto realizzato uno strumento semplificato, ma comunque completo e coerente con gli standard di riferimento, per consentire alle scuole di effettuare il percorso di elaborazione di un'analisi ambientale iniziale. Lo strumento consiste essenzialmente in schede che, una volta compilate

  20. Role of Thr399Ile and Asp299Gly polymorphisms of toll-like receptor-4 gene in acute dental abscess.

    Miri-Moghaddam, Ebrahim; Farhad-Mollashahi, Narges; Baghaee, Elnaz; Bazi, Ali; Garme, Yasaman

    2017-02-01

    Apical Periodontitis (AP) is an inflammatory disease that affects the tissues surrounding the root end of a tooth. The disease which is caused by endodontic infections presents in different clinical ways including development of an acute abscess. Recent studies have provided information suggesting role of a multitude of factors in pathogenesis of acute apical abscess (AAA). In this case-control study, our goal was to evaluate the frequency and potential role of two common polymorphisms of toll like receptor-4 (TLR-4) gene; Thr399Ile (1196 C>T) and Asp299Gly (+896 A>G), in 50 patients with AAA as cases and 50 patients with asymptomatic apical periodontitis (AAP) as controls. Saliva sample containing mucosal epithelial cells was used for DNA extraction. Polymorphisms were detected by Tetra-ARMS (Amplification Refractory Mutation System) PCR method. Statistical analyses were carried out in SPSS 21 software. Homozygous wild type (CC) and heterozygous (CT) genotypes of Thr399Ile polymorphism were detected in 84% and 16% of AAA patients respectively. In controls, respective ratios were 94% (CC) and 6% (CT). Observed difference was not statistically significant ( P >0.05) for distribution of these genotypes. The mutant homozygous (TT) genotype of this polymorphism was identified in neither of the participants. Overall, T allele frequency was obtained 8% in AAA and 3% in AAP (OR=2.6, 95% CI; 0. 6-10.6, p >0.05). For Asp299Gly polymorphism, no individual was detected with the mutant allele in case or control groups. Our results indicated a possible role for Thr399Ile polymorphism in acute presentations of abscess in AAA. However, the impact of this polymorphism needs to be more assessed in future studies. Key words: Genetic polymorphism, periapical abscess, periapical periodontitis, toll-like receptor 4.

  1. Gli effetti delle imposte sulla convenienza a detenere titoli pubblici in Italia (1974-1986. (The effects of taxes on the yields on government bonds in Italy

    A. DI MAJO

    2013-12-01

    Full Text Available Nel 1974 l'Italia ha emanato una riforma fiscale che differenzia il trattamento fiscale per i detentori di obbligazioni a seconda che essi siano imprese o privati cittadini. A sua volta, il " reddito fisso " attività finanziarie sono state distinte in base alla natura dell'emittente , pubblico o privato . Il presente lavoro tenta di tracciare e valutare l'impatto di questa riforma . In particolare, le variazioni di rendimento sui titoli di Stato viene analizzata per gli anni successivi all'introduzione della riforma fino alla sua modifica più recente , realizzato nel mese di settembre 1986. a tal fine , gli autori riassumono la modifica apportata nel codice fiscale prima quantificazione del loro impatto , soprattutto con riferimento ai conti profitti e perdite di credito istituzioni. Alcune indicazioni sugli effetti della più recente modifica del trattamento fiscale degli interessi sono anche descritti In 1974 Italy enacted a tax reform that differentiated the tax treatment for bond holders according to whether they are businesses or private individuals. In turn, "fixed income” financial activities were distinguished based on the nature of the issuer, public or private. The present work attempts to trace and evaluate the impact of this reform. In particular, the changes in yield on government bonds is analysed for the years following the introduction of the reform up to its most recent change, made in September 1986. To this end, the authors summarise the change made in the tax code before quantifying their impact, mainly with reference to the profit and loss accounts of credit institutions. Some indications on the effects of the most recent change in the tax treatment of interest are also outlined.JEL: H20

  2. Environmental analysis applied to schools. Methodologies for data acquisition; Analisi ambientale iniziale per gli istituti d'istruzione. Metodologia per l'acquisizione dei dati

    Andriola, L; Ceccacci, R [ENEA, Div. Caratterizzazione dell' Ambiente e del Territorio, Centro Ricerche Casaccia, Rome (Italy)

    2001-07-01

    The environment analysis is the basis of environmental management for organizations and it is considered as the first step in EMAS. It allows to identify, deal with the issues and have a clear knowledge on environmental performances of organizations. Schools can be included in the organizations. Nevertheless, the complexity of environmental issues and applicable regulations makes very difficult for a school, that wants to implement an environmental management system (EMAS, ISO 14001, etc.), to face this first step. So, it has been defined an instrument, that is easy but complete and coherent with reference standard, to let schools choose their process for elaborating the initial environmental revue. This instrument consists, essentially, in cards that, if completed, facilitate the drafting of the environmental analysis report. [Italian] L'analisi ambientale iniziale costituisce il presupposto di qualsiasi azione di gestione ambientale da parte di un'organizzazione ed e' esplicitamente prevista, come prima tappa, in EMAS. Essa consente infatti di individuare ed approfondire le problematiche ambientali e di fotografare, per mezzo di riferimenti oggettivi, le prestazioni ambientali di un'organizzazione, e quindi anche di una scuola, in modo da poter stabilire e controllare nel tempo gli obiettivi di miglioramento. Tuttavia la complessita' delle problematiche ambientali e delle relative normative di riferimento rende estremamente difficoltoso, per un istituto scolastico che intenda intraprendere un percorso di gestione ambientale comunque finalizzato (EMAS, ISO 14001, Comunicazione ambientale, ecc.) compiere questo passo iniziale. E' stato pertanto realizzato uno strumento semplificato, ma comunque completo e coerente con gli standard di riferimento, per consentire alle scuole di effettuare il percorso di elaborazione di un'analisi ambientale iniziale. Lo strumento consiste essenzialmente in schede che, una volta compilate, faciliteranno la stesura del rapporto di

  3. Impact of engineered Streptococcus thermophilus trains overexpressing glyA gene on folic acid and acetaldehyde production in fermented milk Impacto de linhagens de Streptococcus thermophilus com aumento da expressão do gene glyA na produção de ácido folico e acetaldeído em leite fermentado

    Ana Carolina Sampaio Dória Chaves

    2003-11-01

    Full Text Available The typical yogurt flavor is caused by acetaldehyde produced through many different pathways by the yogurt starter bacteria L. bulgaricus and S. thermophilus. The attention was focused on one specific reaction for acetaldehyde and folic acid formation catalyzed by serine hydroxymethyltransferase (SHMT, encoded by the glyA gene. In S. thermophilus, this enzyme SHMT also plays the typical role of the enzyme threonine aldolase (TA that is the interconvertion of threonine into glycine and acetaldehyde. The behavior of engineered S. thermophilus strains in milk fermentation is described, folic acid and acetaldehyde production were measured and pH and counts were followed. The engineered S. thermophilus strains StA2305 and StB2305, have the glyA gene (encoding the enzyme serine hydroxymethyltransferase overexpressed. These engineered strains showed normal growth in milk when it was supplemented with Casitione. When they were used in milk fermentation it was observed an increase in folic acid and in acetaldehyde production by StA2305 and for StB2305 it was noticed a significative increase in folic acid formation.O acetaldeído, responsável pelo sabor e aroma característicos de iogurte, é produzido por diferentes vias metabólicas pelas bactérias lácticas: Streptococcus thermophilus (S. thermophilus e Lactobacillus delbrueckii subsp. bulgaricus (L. bulgaricus. Neste trabalho, a atenção foi focada especificamente na reação para a formação de acetaldeído e de ácido fólico, catalisada pela enzima serina hidroximetil transferase (SHMT, codificada pelo gene glyA. A enzima SHMT catalisa diversas reações e, no caso da bactéria S. thermophilus, ela exerce também a atividade característica da enzima treonina aldolase (TA, definida como a interconversão do aminoácido treonina em glicina e acetaldeído. Foram construídas linhagens de S. thermophilus (StA2305 e StB2305 com super expressão do gene glyA. Estas linhagens modificadas apresentaram

  4. Effective Biotransformation of Ethyl 4-Chloro-3-Oxobutanoate into Ethyl (S)-4-Chloro-3-Hydroxybutanoate by Recombinant E. coli CCZU-T15 Whole Cells in [ChCl][Gly]-Water Media.

    Dai, Yong; Huan, Bin; Zhang, Hai-Sheng; He, Yu-Cai

    2017-04-01

    To increase the biocatalytic activity of Escherichia coli CCZU-T15 whole cells, choline chloride/glycerol ([ChCl][Gly]) was firstly used as biocompatible solvent for the effective biotransformation of ethyl 4-chloro-3-oxobutanoate (COBE) into ethyl (S)-4-chloro-3-hydroxybutanoate [(S)-CHBE]. Furthermore, L-glutamine (150 mM) was added into [ChCl][Gly]-water ([ChCl][Gly] 12.5 vol%, pH 6.5) media instead of NAD + for increasing the biocatalytic efficiency. To further improve the biosynthesis of (S)-CHBE (>99 % e.e.) by E. coli CCZU-T15 whole cells, Tween-80 (7.5 mM) was also added into this reaction media, and (S)-CHBE (>9 % e.e.) could be effectively synthesized from 2000 and 3000 mM COBE in the yields of 100 and 93.0 % by whole cells of recombinant E. coli CCZU-T15, respectively. TEM image indicated that the cell membrane was permeabilized and lost its integrity and when the cell was exposed to [ChCl][Gly]-water media with Tween-80. Clearly, this bioprocess has high potential for the effective biosynthesis of (S)-CHBE (>99 % e.e.).

  5. Mutation of Gly195 of the ChlH subunit of Mg-chelatase reduces chlorophyll and further disrupts PS II assembly in a Ycf48-deficient strain of Synechocystis sp. PCC 6803

    Tim Crawford

    2016-07-01

    Full Text Available Biogenesis of the photosystems in oxygenic phototrophs requires co-translational insertion of chlorophyll a. The first committed step of chlorophyll a biosynthesis is the insertion of a Mg2+ ion into the tetrapyrrole intermediate protoporphyrin IX, catalyzed by Mg-chelatase. We have identified a Synechocystis sp. PCC 6803 strain with a spontaneous mutation in chlH that results in a Gly195 to Glu substitution in a conserved region of the catalytic subunit of Mg-chelatase. Mutant strains containing the ChlH Gly195 to Glu mutation were generated using a two-step protocol that introduced the chlH gene into a putative neutral site in the chromosome prior to deletion of the native gene. The Gly195 to Glu mutation resulted in strains with decreased chlorophyll a. Deletion of the PS II assembly factor Ycf48 in a strain carrying the ChlH Gly195 to Glu mutation did not grow photoautotrophically. In addition, the ChlH-G195E:ΔYcf48 strain showed impaired PS II activity and decreased assembly of PS II centers in comparison to a ΔYcf48 strain. We suggest decreased chlorophyll in the ChlH-G195E mutant provides a background to screen for the role of assembly factors that are not essential under optimal growth conditions.

  6. Influence of turn (or fold) and local charge in fragmentation of the peptide analogue molecule CH3CO-Gly-NH2 following single-photon VUV (118.22 nm) ionization.

    Bhattacharya, Atanu; Bernstein, Elliot R

    2011-10-06

    The radical cationic reactivity of the peptide analogue molecule CH(3)CO-Gly-NH(2) is addressed both experimentally and theoretically. The radical cation intermediate of CH(3)CO-Gly-NH(2) is created by single-photon ionization of this molecule at 118.22 nm (~10.5 eV). The two most stable conformers (C(7) and C(5)) of this molecule exhibit different folds along the backbone: the C(7) conformer has a γ-turn structure, and the C(5) conformer has a β-strand structure. The experimental results show that the radical cation intermediate of CH(3)CO-Gly-NH(2) dissociates and generates a fragment-ion signal at 73 amu that is observed through TOFMS. Theoretical results show how the fragment-ion signal at 73 amu is generated by only one conformer of CH(3)CO-Gly-NH(2) (C(7)) and how local charge and specific hydrogen bonding in the molecule influence fragmentation of the radical cation intermediate of CH(3)CO-Gly-NH(2). The specific fold of the molecule controls fragmentation of this reactive radical cation intermediate. Whereas the radical cation of the C(7) conformer dissociates through a hydrogen-transfer mechanism followed by HNCO elimination, the radical cation of the C(5) conformer does not dissociate at all. CASSCF calculations show that positive charge in the radical cationic C(7) conformer is localized at the NH(2)CO moiety of the molecular ion. This site-specific localization of the positive charge enhances the acidity of the terminal NH(2) group, facilitating hydrogen transfer from the NH(2) to the COCH(3) end of the molecular ion. Positive charge in the C(5) conformer of the CH(3)CO-Gly-NH(2) radical cation is, however, localized at the COCH(3) end of the molecular ion, and this conformer does not have enough energy to surmount the energy barrier to dissociation on the ion potential energy surface. CASSCF results show that conformation-specific localization of charge in the CH(3)CO-Gly-NH(2) molecular ion occurs as a result of the different hydrogen

  7. Obesity is associated with the Arg389Gly ADRB1 but not with the Trp64Arg ADRB3 polymorphism in children from San Luis PotosÍ and León, México.

    Aradillas-Garc X Cd, Celia; Cruz, Miguel; Pérez-Luque, Elva; Garay-Sevilla, María E; Malacara, Juan M; R, Aduna; Peralta, Jesús; Burguete-García, Ana; Alegría-Torres, Jorge A

    2016-10-17

    This research was designed to analyze the possible associations of Arg389Gly ADRB1 and Trp64Arg ADRB3 polymorphisms in children with obesity. A cross-sectional study included 1,046 school-age Mexican participants (6-12 years old) from the cities of San Luis PotosÍ and León. Children were classified as non-obese or obese according to their body mass index (BMI) percentile; obese children had a BMI≥95th percentile for sex and age. Biochemical data were collected. Polymorphisms were detected using TaqMan qPCR assay. A logistic regression analysis was used to calculate the risk of obesity based on genotypes. Differences were found between groups where obese children had a significant increase in systolic and diastolic blood pressure, fasting plasma glucose, insulin, HOMA-IR, LDL-cholesterol, triglycerides, and lower HDL-cholesterol compared with the normal weight group (P<0.05). The distribution of allele frequency in the population was Arg= 87.4 and Gly= 12.6 (Hardy Weinberg equilibrium c 2 = 3.16 , P = 0.07 ); Trp= 81.5 and Arg= 18.5 (Hardy Weinberg equilibrium c 2 = 2.2, P = 0.14 ) for ADRB1 and ADRB3, respectively. Even though no different frequencies of Arg389Gly polymorphism between groups were found (P = 0.08), children carriers of one Gly389 ADRB1 allele had a risk for obesity of OR=1.40 (95%CI, 1.03-1.90, P = 0.03) after adjustment for age and gender. No other association was found for Trp64Arg ADRB3 polymorphism. Only the Arg389Gly ADRB1 polymorphism was associated with risk for obesity in Mexican children.

  8. Una «tragedia in forma di romanzo»? Teatralità e intertestualità pirandelliana ne Gli indifferenti di Alberto Moravia / A «tragedia in forma di romanzo» (tragedy in the form of a novel? Theatricality and Pirandello’s inrtextuality in “Gli Indifferenti” by Alberto Moravia.

    Agnese Marasca

    2015-06-01

    Gli indifferenti by Moravia is a theatrical novel both in the form for the use of techniques and procedures typical of drama, and in the content for the presence of subjects and figures which refer to the theater. The dimension of theatrics in the novel is also closely related to the intertextuality with Pirandello, from whom philosophical imaginary the romance takes a lot of suggestions, as first the mask and the staging social. But what deeply connects Gli indifferenti with the work of Pirandello is the theme of the impossibility of the tragedy, that is the first cause of Michele’s ineptitude in life. The loss of the tragic sense prevents the realization of the oedipal drama, because the tragic of the novel remains a purely external fact. The absence of any tragical assumption is clarified thanks to the comparison of the plot and the characters with the ones of Sei personaggi in cerca d’autore, with which Moravia’s novel has dense and hidden relations of intertextuality.

  9. Catestatin Gly364Ser Variant Alters Systemic Blood Pressure and the Risk for Hypertension in Human Populations via Endothelial Nitric Oxide Pathway.

    Kiranmayi, Malapaka; Chirasani, Venkat R; Allu, Prasanna K R; Subramanian, Lakshmi; Martelli, Elizabeth E; Sahu, Bhavani S; Vishnuprabu, Durairajpandian; Kumaragurubaran, Rathnakumar; Sharma, Saurabh; Bodhini, Dhanasekaran; Dixit, Madhulika; Munirajan, Arasambattu K; Khullar, Madhu; Radha, Venkatesan; Mohan, Viswanathan; Mullasari, Ajit S; Naga Prasad, Sathyamangla V; Senapati, Sanjib; Mahapatra, Nitish R

    2016-08-01

    Catestatin (CST), an endogenous antihypertensive/antiadrenergic peptide, is a novel regulator of cardiovascular physiology. Here, we report case-control studies in 2 geographically/ethnically distinct Indian populations (n≈4000) that showed association of the naturally-occurring human CST-Gly364Ser variant with increased risk for hypertension (age-adjusted odds ratios: 1.483; P=0.009 and 2.951; P=0.005). Consistently, 364Ser allele carriers displayed elevated systolic (up to ≈8 mm Hg; P=0.004) and diastolic (up to ≈6 mm Hg; P=0.001) blood pressure. The variant allele was also found to be in linkage disequilibrium with other functional single-nucleotide polymorphisms in the CHGA promoter and nearby coding region. Functional characterization of the Gly364Ser variant was performed using cellular/molecular biological experiments (viz peptide-receptor binding assays, nitric oxide [NO], phosphorylated extracellular regulated kinase, and phosphorylated endothelial NO synthase estimations) and computational approaches (molecular dynamics simulations for structural analysis of wild-type [CST-WT] and variant [CST-364Ser] peptides and docking of peptide/ligand with β-adrenergic receptors [ADRB1/2]). CST-WT and CST-364Ser peptides differed profoundly in their secondary structures and showed differential interactions with ADRB2; although CST-WT displaced the ligand bound to ADRB2, CST-364Ser failed to do the same. Furthermore, CST-WT significantly inhibited ADRB2-stimulated extracellular regulated kinase activation, suggesting an antagonistic role towards ADRB2 unlike CST-364Ser. Consequently, CST-WT was more potent in NO production in human umbilical vein endothelial cells as compared with CST-364Ser. This NO-producing ability of CST-WT was abrogated by ADRB2 antagonist ICI 118551. In conclusion, CST-364Ser allele enhanced the risk for hypertension in human populations, possibly via diminished endothelial NO production because of altered interactions of CST-364Ser

  10. Toll-like receptor 4 Asp299Gly and Thr399Ile polymorphisms and typhoid susceptibility in Asian Malay population in Malaysia.

    Bhuvanendran, Saatheeyavaane; Hussin, Hani M; Meran, Lila P; Anthony, Amy A; Zhang, Leilei; Burch, Lauranell H; Phua, Kia K; Ismail, Asma; Balaram, Prabha

    2011-09-01

    Typhoid fever is a major health problem with frequent outbreaks in Kelantan, Malaysia. Prevalence of TLR4 gene polymorphisms varies with ethnic groups (0-20%) and predisposean individual to gram-negative infections. The prevalence rate of TLR4 Asp299Gly and Thr399lle polymorphisms in the Malay population or the influence of these on typhoid fever susceptibility is not yet reported. 250 normal and 304 susceptible Malay individuals were investigated for these polymorphisms using allele-specific PCR and analysed for its association with typhoid fever susceptibility. The total prevalence of polymorphisms in the normal population was 4.8% in comparison to 12.5% in the susceptible population (p = 0.002). An increased frequency of both polymorphisms was observed in the susceptible population (p population and suggests that these polymorphisms confer a higher risk for typhoid, infection. The higher incidence of typhoid fever in Kelantan could be attributed to the higher percentage of Malays (95%) in this state. In order to reduce the incidence of this disease, people with these polymorphisms, can be prioritised for prophylactic strategies. Copyright © 2011 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  11. Effects of ionizing radiation and pretreatment with [D-Leu6,des-Gly10] luteinizing hormone-releasing hormone ethylamide on developing rat ovarian follicles

    Jarrell, J.; YoungLai, E.V.; McMahon, A.; Barr, R.; O'Connell, G.; Belbeck, L.

    1987-01-01

    To assess the effects of a gonadotropin-releasing hormone agonist, [D-Leu6,des-Gly10] luteinizing hormone-releasing hormone ethylamide, in ameliorating the damage caused by ionizing radiation, gonadotropin-releasing hormone agonist was administered to rats from day 22 to 37 of age in doses of 0.1, 0.4, and 1.0 microgram/day or vehicle and the rats were sacrificed on day 44 of age. There were no effects on estradiol, progesterone, luteinizing, or follicle-stimulating hormone, nor an effect on ovarian follicle numbers or development. In separate experiments, rats treated with gonadotropin-releasing hormone agonist in doses of 0.04, 0.1, 0.4, or 1.0 microgram/day were either irradiated or sham irradiated on day 30 and all groups sacrificed on day 44 of age. Irradiation produced a reduction in ovarian weight and an increase in ovarian follicular atresia. Pretreatment with the agonist prevented the reduction in ovarian weight and numbers of primordial and preantral follicles but not healthy or atretic antral follicles. Such putative radioprotection should be tested on actual reproductive performance

  12. Sox2, Tlx, Gli3, and Her9 converge on Rx2 to define retinal stem cells in vivo

    Reinhardt, Robert; Centanin, Lázaro; Tavhelidse, Tinatini; Inoue, Daigo; Wittbrodt, Beate; Concordet, Jean-Paul; Martinez-Morales, Juan Ramón; Wittbrodt, Joachim

    2015-01-01

    Transcriptional networks defining stemness in adult neural stem cells (NSCs) are largely unknown. We used the proximal cis-regulatory element (pCRE) of the retina-specific homeobox gene 2 (rx2) to address such a network. Lineage analysis in the fish retina identified rx2 as marker for multipotent NSCs. rx2-positive cells located in the peripheral ciliary marginal zone behave as stem cells for the neuroretina, or the retinal pigmented epithelium. We identified upstream regulators of rx2 interrogating the rx2 pCRE in a trans-regulation screen and focused on four TFs (Sox2, Tlx, Gli3, and Her9) activating or repressing rx2 expression. We demonstrated direct interaction of the rx2 pCRE with the four factors in vitro and in vivo. By conditional mosaic gain- and loss-of-function analyses, we validated the activity of those factors on regulating rx2 transcription and consequently modulating neuroretinal and RPE stem cell features. This becomes obvious by the rx2-mutant phenotypes that together with the data presented above identify rx2 as a transcriptional hub balancing stemness of neuroretinal and RPE stem cells in the adult fish retina. PMID:25908840

  13. Continuing evolution of canine parvovirus in China: Isolation of novel variants with an Ala5Gly mutation in the VP2 protein.

    Wang, Jianke; Lin, Peng; Zhao, Hang; Cheng, Yuening; Jiang, Zhong; Zhu, Hongwei; Wu, Hua; Cheng, Shipeng

    2016-03-01

    Canine parvovirus (CPV) type 2c is a new antigenic variant of CPV-2. Since the year 2000 it has spread to several countries, causing severe hemorrhagic enteritis in dogs. In 2014 and 2015, 58 fecal samples were collected from dogs in Beijing with suspected CPV infection. Regardless of the vaccination status of the dogs, 43 samples were found positive for CPV according to PCR results; i.e., 18, 7, and 18 respectively belonged to antigenic types new CPV-2a, new CPV-2b, and CPV-2c. A phylogenetic tree based on their VP2 gene sequences indicated that the Chinese CPV-2c strains form a separate cluster. In addition to synonymous mutations, the CPV-2c strains also contain a unique coding mutation in VP2 that leads to glycine at residue 5, instead of the highly conserved alanine at this position in all other CPV-2c strains sequenced to date. Using F81 cells, several novel isolates of CPV-2c, each with the Ala5Gly mutation, were obtained. One of these was used to infect experimentally beagle dogs, which subsequently developed the typical clinical symptoms of CPV infection. Hence, it appears that CPV-2c is still evolving in China, a finding that warrants continuous surveying and the eventual adaptation of current vaccines. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Sedimentology and biostratigraphy of the Early-Middle Miocene transition in NW Transylvanian Basin (Pâglişa and Dej sections

    Claudia Beldean

    2013-01-01

    Full Text Available The Early - Middle Miocene transition can be studied in several sections from the north-western Transylvanian Basin, across the boundary between the Hida and Dej Formations. The Pâglişa and Râpa Dracului (Dej locally outcrop sections offer a very good record of the changes in the sedimentary facies and micropaleontological content across the boundary. The lower parts of the sections display the upper Hida Formation with prodelta fine grained sediments covered by coarser grained sediments related to a delta front progradation. Four sedimentary facies associations have been separated based on grain size and associated sedimentary structures. Micropaleontological assemblages are moderately to poorly preserved, with a low abundance and diversity. Planktonic Streptochilus pristinum is the main taxon associated with rare (< 10% calcareous benthic foraminifera (Bulimina, Bolivina, Cibibicidoides. The Dej Formation consists of stacked channel lag deposits associated with lateral accretion bars and crevasse splays followed by fall-out tuffs and redeposited low density tuffites interbedded with low density fine siliciclastics. The deposits of Dej Formation from the investigated areas were separated in four facies associations based on grain size/petrography and primary sedimentary structures. Micropaleontological assemblages contain typical early Badenian planktonic foraminifera (species of Orbulina, Praeorbulina, Globigerinoides, and Globorotalia associated to the main phase of the marine transgresion from the beginning of the Middle Miocene.

  15. Over-expression of Arabidopsis thaliana SFD1/GLY1, the gene encoding plastid localized glycerol-3-phosphate dehydrogenase, increases plastidic lipid content in transgenic rice plants.

    Singh, Vijayata; Singh, Praveen Kumar; Siddiqui, Adnan; Singh, Subaran; Banday, Zeeshan Zahoor; Nandi, Ashis Kumar

    2016-03-01

    Lipids are the major constituents of all membranous structures in plants. Plants possess two pathways for lipid biosynthesis: the prokaryotic pathway (i.e., plastidic pathway) and the eukaryotic pathway (i.e., endoplasmic-reticulum (ER) pathway). Whereas some plants synthesize galactolipids from diacylglycerol assembled in the plastid, others, including rice, derive their galactolipids from diacylglycerols assembled by the eukaryotic pathway. Arabidopsis thaliana glycerol-3-phosphate dehydrogenase (G3pDH), coded by SUPPRESSOR OF FATTY ACID DESATURASE 1 (SFD1; alias GLY1) gene, catalyzes the formation of glycerol 3-phosphate (G3p), the backbone of many membrane lipids. Here SFD1 was introduced to rice as a transgene. Arabidopsis SFD1 localizes in rice plastids and its over-expression increases plastidic membrane lipid content in transgenic rice plants without any major impact on ER lipids. The results suggest that over-expression of plastidic G3pDH enhances biosynthesis of plastid-localized lipids in rice. Lipid composition in the transgenic plants is consistent with increased phosphatidylglycerol synthesis in the plastid and increased galactolipid synthesis from diacylglycerol produced via the ER pathway. The transgenic plants show a higher photosynthetic assimilation rate, suggesting a possible application of this finding in crop improvement.

  16. Ile-1781-Leu and Asp-2078-Gly Mutations in ACCase Gene, Endow Cross-resistance to APP, CHD, and PPZ in Phalaris minor from Mexico

    Hugo Cruz-Hipolito

    2015-09-01

    Full Text Available Herbicides that inhibit acetyl coenzyme A carboxylase (ACCase are commonly used in Mexico to control weedy grasses such as little seed canarygrass (Phalaris minor. These herbicides are classified into three major families (ariloxyphenoxypropionates (APP, cyclohexanodiones (CHD, and, recently, phenylpyrazolines (PPZ. In this work, the resistance to ACCase (APP, CHD, and PPZ inhibiting herbicides was studied in a biotype of Phalaris minor (P. minor from Mexico, by carrying out bioassays at the whole-plant level and investigating the mechanism behind this resistance. Dose-response and ACCase in vitro activity assays showed cross-resistance to all ACCase herbicides used. There was no difference in the absorption, translocation, and metabolism of the 14C-diclofop-methyl between the R and S biotypes. The PCR generated CT domain fragments of ACCase from the R biotype and an S reference were sequenced and compared. The Ile-1781-Leu and Asp-2078-Gly point mutations were identified. These mutations could explain the loss of affinity for ACCase by the ACCase-inhibing herbicides. This is the first report showing that this substitution confers resistance to APP, CHD, and PPZ herbicides in P. minor from Mexico. The mutations have been described previously only in a few cases; however, this is the first study reporting on a pattern of cross-resistance with these mutations in P. minor. The findings could be useful for better management of resistant biotypes carrying similar mutations.

  17. Fisica del nucleo stelle di neutroni

    Bernardini, Carlo

    1982-01-01

    Studio sistematico delle masse nucleari ; energia di legame ; raggi nucleari dai nuclei speculari ; il nucleo in approssimazione di Thomas-Fermi ; introduzione allo scattering : caratterizzazione cinematica degli urti ; la sezione d'urto ; il fattore di forma dei nuclei ; gli atomi u-mesici ; proprieta elettromagnetiche statiche dei nuclei (momenti di multipolo) ; momento di quadrupolo elettrico dei nuclei ; momento d dipolo magnetico dei nuclei ; metodo delle risonanze magnetiche nucleari (NMR) ; misura del fattore giromagnetico e del momento di quadrupolo ; modelli nucleari a particelle indipendenti, il modello a shell ; l'interazione spin-orbita ; il modello a shell e le linee di Schmidt per il momento magnetico dei nuclei con "A" dispari ; nuclei non sferici (deformati) ; stelle di neutroni.

  18. The expression of Hedgehog genes (Ihh, Dhh) and Hedgehog target genes (Ptc1, Gli1, Coup-TfII) is affected by estrogenic stimuli in the uterus of immature female rats.

    Katayama, Seiichi; Ashizawa, Koji; Gohma, Hiroshi; Fukuhara, Tadahiro; Narumi, Kazunori; Tsuzuki, Yasuhiro; Tatemoto, Hideki; Nakada, Tadashi; Nagai, Kenji

    2006-12-15

    The objective of this study was to investigate the effects of estrogen receptor (ER) agonists and an ER antagonist on the expression of Hedgehog genes (Indian hedgehog: Ihh; Desert hedgehog: Dhh) and Hedgehog target genes (Patched 1: Ptc1; glioma-associated oncogene homolog 1: Gli1; chicken ovalbumin upstream promoter transcription factor II: Coup-TfII) in the rat uterus. Immature female rats were administered once with 17alpha-ethynyl estradiol (EE, an ER agonist), propyl pyrazole triole (PPT, an ERalpha-selective agonist), diarylpropionitrile (DPN, an ERbeta-selective agonist), or ICI 182,780 (an ER antagonist). Expression of mRNA for Ihh, Dhh, and Ptc1 was dose-dependently downregulated by EE in the uterus of immature rats, mediated by ER as confirmed by coadministration of ICI 182,780. The mRNA expression levels of Ptc1, Gli1, and Coup-TfII were simultaneously downregulated during the period in which the mRNA expression levels of Ihh and Dhh were downregulated in the uterus after administration of EE. PPT downregulated the transcription of Ihh, Dhh, Ptc1, Gli1, and Coup-TfII, indicating that expression of these genes was regulated by the ERalpha-dependent pathway. DPN also downregulated the transcription of Ihh and Dhh, although the effect was weaker than that of PPT, indicating that the regulation of uterine Ihh and Dhh transcription was also affected by the ERbeta-dependent pathway. These results suggest that the expression of Hedgehog genes (Ihh, Dhh) and Hedgehog target genes (Ptc1, Gli1, Coup-TfII) is affected by estrogenic stimuli in the uterus of immature female rats.

  19. The expression of Hedgehog genes (Ihh, Dhh) and Hedgehog target genes (Ptc1, Gli1, Coup-TfII) is affected by estrogenic stimuli in the uterus of immature female rats

    Katayama, Seiichi; Ashizawa, Koji; Gohma, Hiroshi; Fukuhara, Tadahiro; Narumi, Kazunori; Tsuzuki, Yasuhiro; Tatemoto, Hideki; Nakada, Tadashi; Nagai, Kenji

    2006-01-01

    The objective of this study was to investigate the effects of estrogen receptor (ER) agonists and an ER antagonist on the expression of Hedgehog genes (Indian hedgehog: Ihh; Desert hedgehog: Dhh) and Hedgehog target genes (Patched 1: Ptc1; glioma-associated oncogene homolog 1: Gli1; chicken ovalbumin upstream promoter transcription factor II: Coup-TfII) in the rat uterus. Immature female rats were administered once with 17α-ethynyl estradiol (EE, an ER agonist), propyl pyrazole triole (PPT, an ERα-selective agonist), diarylpropionitrile (DPN, an ERβ-selective agonist), or ICI 182,780 (an ER antagonist). Expression of mRNA for Ihh, Dhh, and Ptc1 was dose-dependently downregulated by EE in the uterus of immature rats, mediated by ER as confirmed by coadministration of ICI 182,780. The mRNA expression levels of Ptc1, Gli1, and Coup-TfII were simultaneously downregulated during the period in which the mRNA expression levels of Ihh and Dhh were downregulated in the uterus after administration of EE. PPT downregulated the transcription of Ihh, Dhh, Ptc1, Gli1, and Coup-TfII, indicating that expression of these genes was regulated by the ERα-dependent pathway. DPN also downregulated the transcription of Ihh and Dhh, although the effect was weaker than that of PPT, indicating that the regulation of uterine Ihh and Dhh transcription was also affected by the ERβ-dependent pathway. These results suggest that the expression of Hedgehog genes (Ihh, Dhh) and Hedgehog target genes (Ptc1, Gli1, Coup-TfII) is affected by estrogenic stimuli in the uterus of immature female rats

  20. Structure-activity relationship of linear peptide Bu-His6-DPhe7-Arg8-Trp9-Gly10-NH2 at the human melanocortin-1 and -4 receptors: DPhe7 and Trp9 substitution.

    Danho, Waleed; Swistok, Joseph; Cheung, Adrian Wai-Hing; Kurylko, Grazyna; Franco, Lucia; Chu, Xin-Jie; Chen, Li; Yagaloff, Keith

    2003-02-24

    A series of pentapeptides, based on hMC4R pentapeptide agonist (Bu-His(6)-DPhe(7)-Arg(8)-Trp(9)-Gly(10)-NH(2)), was prepared in which either DPhe(7) or Trp(9) residue was systematically substituted. A number of interesting DPhe surrogates (D-Thi, D-3-CF(3)Phe, D-2-Nal and D-3,4-diClPhe) as well as Trp surrogates (2-Nal and Bta) were identified in this study.

  1. “Economic heritage impact assessment” come strumento per valutare gli impatti dei grandi progetti di riqualificazione delle città costiere siti Unesco. Il caso studio di Torre Annunziata nel golfo di Napoli

    Mariarosaria Angrisano

    2015-12-01

    Full Text Available Quest’articolo parte dallo studio delle città portuali storiche, siti UNESCO, che oggi affrontano la sfida della riqualificazione dei waterfront urbani attraverso la conservazione del patrimonio culturale e paesaggistico. Le raccomandazioni sull’ “Historic Urban Landscape”, e gli strumenti operativi promossi dall’ICOMOS “Guidance on Heritage Impact Assessments for Cultural World Heritage Properties” del 2011, si configurano come le più recenti disposizioni concernenti la conservazione, protezione e valorizzazione del patrimonio culturale (Fusco Girard, 2010. La Guida ICOMOS è stata considerata lo strumento più adatto per valutare gli impatti dei grandi progetti di riqualificazione dei waterfront urbani sul patrimonio culturale. Per comprendere al meglio tale strumento è stata fatta un’applicazione per valutare gli impatti del Grande Progetto Pompei sul waterfront di Torre Annunziata, città costiera nel Golfo di Napoli sito UNESCO. A questa fase di analisi segue una riflessione su come migliorare l’Heritage Impact Assessment, come strumento capace di valutare non soltanto gli impatti culturali ma anche quelli economici, per parlare di “economia del patrimonio culturale”. La proposta ultima è quella di affiancare al processo di “Heritage Impact Assessment” una valutazione degli impatti economici, attraverso un’analisi costi-benefici, per quantificare in termini monetari la convenienza degli investimenti nella conservazione del patrimonio storico urbano delle città costiere, proponendo una “Economic Heritage Impact Assessment” (EHIA. Tale metodo, elaborato dall’autrice all’interno della tesi di dottorato “Il paesaggio storico urbano delle città costiere: sfide e opportunità. Il caso Torre Annunziata”, offre la possibilità di superare la sola valutazione degli impatti culturali proposta dall’ICOMOS.

  2. Gli2a protein localization reveals a role for Iguana/DZIP1 in primary ciliogenesis and a dependence of Hedgehog signal transduction on primary cilia in the zebrafish

    van Eeden Freek

    2010-04-01

    Full Text Available Abstract Background In mammalian cells, the integrity of the primary cilium is critical for proper regulation of the Hedgehog (Hh signal transduction pathway. Whether or not this dependence on the primary cilium is a universal feature of vertebrate Hedgehog signalling has remained contentious due, in part, to the apparent divergence of the intracellular transduction pathway between mammals and teleost fish. Results Here, using a functional Gli2-GFP fusion protein, we show that, as in mammals, the Gli2 transcription factor localizes to the primary cilia of cells in the zebrafish embryo and that this localization is modulated by the activity of the Hh pathway. Moreover, we show that the Igu/DZIP1protein, previously implicated in the modulation of Gli activity in zebrafish, also localizes to the primary cilium and is required for its proper formation. Conclusion Our findings demonstrate a conserved role of the primary cilium in mediating Hedgehog signalling activity across the vertebrate phylum and validate the use of the zebrafish as a representative model for the in vivo analysis of vertebrate Hedgehog signalling.

  3. HECT E3 Ubiquitin Ligase Itch Functions as a Novel Negative Regulator of Gli-Similar 3 (Glis3 Transcriptional Activity.

    Gary T ZeRuth

    Full Text Available The transcription factor Gli-similar 3 (Glis3 plays a critical role in the generation of pancreatic ß cells and the regulation insulin gene transcription and has been implicated in the development of several pathologies, including type 1 and 2 diabetes and polycystic kidney disease. However, little is known about the proteins and posttranslational modifications that regulate or mediate Glis3 transcriptional activity. In this study, we identify by mass-spectrometry and yeast 2-hybrid analyses several proteins that interact with the N-terminal region of Glis3. These include the WW-domain-containing HECT E3 ubiquitin ligases, Itch, Smurf2, and Nedd4. The interaction between Glis3 and the HECT E3 ubiquitin ligases was verified by co-immunoprecipitation assays and mutation analysis. All three proteins interact through their WW-domains with a PPxY motif located in the Glis3 N-terminus. However, only Itch significantly contributed to Glis3 polyubiquitination and reduced Glis3 stability by enhancing its proteasomal degradation. Itch-mediated degradation of Glis3 required the PPxY motif-dependent interaction between Glis3 and the WW-domains of Itch as well as the presence of the Glis3 zinc finger domains. Transcription analyses demonstrated that Itch dramatically inhibited Glis3-mediated transactivation and endogenous Ins2 expression by increasing Glis3 protein turnover. Taken together, our study identifies Itch as a critical negative regulator of Glis3-mediated transcriptional activity. This regulation provides a novel mechanism to modulate Glis3-driven gene expression and suggests that it may play a role in a number of physiological processes controlled by Glis3, such as insulin transcription, as well as in Glis3-associated diseases.

  4. The immunoprofile of odontogenic keratocyst (keratocystic odontogenic tumor) that includes expression of PTCH, SMO, GLI-1 and bcl-2 is similar to ameloblastoma but different from odontogenic cysts.

    Vered, M; Peleg, O; Taicher, S; Buchner, A

    2009-08-01

    The aggressive biological behavior of odontogenic keratocysts (OKCs), unlike that of other odontogenic cysts, has argued for its recent re-classification as a neoplasm, 'keratocystic odontogenic tumor'. Identification of mutations in the PTCH gene in some of the OKCs that were expected to produce truncated proteins, resulting in loss of control of the cell cycle, provided additional support for OKCs having a neoplastic nature. We investigated the immunohistochemical expression of the sonic hedgehog (SHH) signaling pathway-related proteins, PTCH, smoothened (SMO) and GLI-1, and of the SHH-induced bcl-2 oncoprotein in a series of primary OKC (pOKC), recurrent OKC (rOKC) and nevoid basal cell carcinoma syndrome-associated OKCs (NBCCS-OKCs), and compared them to solid ameloblastomas (SAMs), unicystic ameloblastomas (UAMs), 'orthokeratinized' OKCs (oOKCs), dentigerous cysts (DCs) and radicular cysts (RCs). All studied lesions expressed the SHH pathway-related proteins in a similar pattern. The expression of bcl-2 in OKCs (pOKCs and NBCCS-OKCs) and SAMs was significantly higher than in oOKCs, DCs and RCs (P < 0.001). The present results of the immunoprofile of OKCs (that includes the expression of the SHH-related proteins and the SHH-induced bcl-2 oncoprotein) further support the notion of OKC having a neoplastic nature. As OKCs vary considerably in their biologic behavior, it is suggested that the quality and quantity of interactions between the SHH and other cell cycle regulatory pathways are likely to work synergistically to define the individual phenotype and corresponding biological behavior of this lesion.

  5. Model-free nuclear magnetic resonance study of intermolecular free energy landscapes in liquids with paramagnetic Ln3+ spotlights: theory and application to Arg-Gly-Asp.

    Fries, Pascal H

    2012-01-28

    We propose an easily applicable method for investigating the pair distribution function of a lanthanide Ln(3+) complex LnL (L = ligand) with respect to any solvent or solute molecule A carrying observable nuclear spins. Let r be the distance of Ln(3+) to the observed nuclear spin I. We derive a simple expression of the experimental value of the configurational average of 1/r(6) in terms of longitudinal paramagnetic relaxation (rate) enhancements (PREs) of the spin I measured on a standard high-resolution NMR spectrometer and due to well-chosen concentrations of LnL complexes in which Ln(3+) is a fast-relaxing paramagnetic lanthanide or the slowly-relaxing gadolinium Gd(3+). The derivation is justified in the general case of a molecule A which is by turns in a bound state where it follows the complex and a free state where it moves independently. It rests on the expression of the underlying PRE theory in terms of the angle-dependent pair distribution function of LnL and A. The simplifications of this theory in the high-field regime and under the condition of fast exchange between bound and free states are carefully discussed. We also show that original information on the angle dependence of the molecular pair distribution function can be gained from the measured paramagnetic dipolar shifts induced by complexed fast-relaxing Ln(3+) ions. The method is illustrated by the case study of the anionic Lnttha(3-) = [Ln(3+)(ttha)](3-) (ttha(6-) = triethylene tetraamine hexacetate) complex interacting with the biologically important tripeptide Arg-Gly-Asp (RGD) which carries peripheral ionic groups. The usefulness of an auxiliary reference outer sphere probe solute is emphasized. © 2012 American Institute of Physics

  6. A Clinical Update of the Hb Siirt [β27(B9)Ala→Gly; HBB: c.83C>G] Hemoglobin Variant.

    Cappabianca, Maria Pia; Colosimo, Alessia; Sabatucci, Annalaura; Dainese, Enrico; Di Biagio, Paola; Piscitelli, Roberta; Sarra, Ofelia; Zei, Daniela; Amato, Antonio

    2017-01-01

    We report a clinical update of the hemoglobin (Hb) variant [β27(B9)Ala→Gly; HBB: c.83C>G], named Hb Siirt, that was previously described as a silent variant in a 23-year-old Kurdish female. The patient was also a carrier of the codon 5 (-CT) (HBB: c.17_18delCT) frameshift mutation and of the ααα anti 3.7 triplication. Her initial moderate β-thalassemia intermedia (β-TI) phenotype worsened with time, causing the patient to become a transfusion-dependent subject at the age of ∼40 years. Subsequent molecular characterization of both parents revealed that the Hb Siirt variant was inherited by the mother, while the other two globin alterations (HBB: c.17_18delCT and ααα anti 3.7 triplication) were genetically transmitted by the father. The latter remained a carrier of a mild β-TI phenotype throughout his life, at least until the age of 65 years. We hypothesize that the worsened clinical conditions in the daughter were due to the additional, maternally inherited Hb Siirt variant. However, protein 3D conformational analysis did not seem to reveal substantial overall structural changes. Among the other three described variants [Hb Volga (HBB: c.83C>A), Hb Knossos (HBB: c.82 G>T), Hb Grange-Blanche (HBB: c.83C>T] that are due to nucleotide substitutions at codon 27 of the β-globin gene; only Hb Knossos causes a β + -thalassemia (β + -thal) phenotype.

  7. Gerberto, gli Arabi e Gerusalemme

    Sigismondi, C.

    Gerbert of Aurillac went in Catalonia from 967 to 970, to visit the bishop Atton of Vic, famous for mathematical knowledge, during the apogee of Cordova's reign of the Caliph Al-Hakam II ibn Abd al-Rahman (961-976). He maintained contacts with local personalities like Mirò Bonfill (Bishop of Girona) and Lupitus of Barcelona through letters (both dated 984). When he was pope, Gerbert sent five papal privileges to Catalan dignitaries, showing his attention to this territory. The Caliph's prime minister Al-Mansu'r depredated Barcelona in 985, and conquered Santiago de Compostela in 997, drawing to an end the cultural exchanges between Christian and Arabic world through Mozarabic and Jewish translators. Gerbert learned and after taught and used Indo-Arabic numbers, abaci, astrolabes, monochords and theoretical music. It was possible for pilgrims to reach Jerusalem at Gerbert's times, and a letter written by Gerbert probably for an abbot starting his pilgrimage seems to introduce the first ideas of crusade and indulgences related to offers. The change of attitude of the Al-Andalus' government toward Christianity is probably responsible of these words written in a time very close to the depredations of Barcelona. Jerusalem is also in the index of geographic places in the book of the astrolabe, attributed to Gerbert, as well as Santiago de Compostela. Finally the legend of the Golem and the mass celebrated in the church of Jerusalem in Rome, completes the panorama of Gerbert and the Holy City.

  8. Sostenere le produzioni cinematografiche: vantaggi fiscali e di immagine per gli investitori / Sustaining cinematographic productions: fiscal and image opportunities for investors

    Katia Giusepponi

    2016-05-01

    Full Text Available Questo contributo è focalizzato su organizzazioni non appartenenti al settore cinematografico, osservate nel loro ruolo di investitori esterni. L’importanza del valore collettivo di sostenere le produzioni cineaudiovisive è alla base del presente lavoro. Tuttavia la nostra attenzione è qui rivolta, da un lato, a studiare i vantaggi dell’investimento per l’azienda esterna, dall’altro, ad analizzarne i rischi nella prospettiva di una sempre più chiara consapevolezza di scelta. Dall’analisi di strumenti, dinamiche e dati emergono opportunità rilevanti per l’azienda, insieme a condizioni importanti per utilizzarle con successo. È essenziale che l’organizzazione proceda senza improvvisare, curando, da un lato, un approccio alla pianificazione e alla dettagliata valutazione del progetto, dall’altro, flessibilità e apertura nelle interazioni con gli altri interlocutori, interazioni che devono essere orientate al raggiungimento dell’equilibrio migliore a livello collettivo e individuale. Soprattutto, è necessario che l’azienda proceda nel contesto di un percorso strategico solido e coerente, perché un progetto di film può valorizzare l’identità aziendale ma non risolvere incertezze a questo riguardo.   This contribution focuses on organisations that are not in the cinema industry, studied in their role as external investors. The importance of the collective value of sustaining cine audio visual productions is at the basis of this work. Nonetheless, our attention is oriented, on one hand, to study the investment advantages for the external organization, on the other, to analyse the risks of this investment in the perspective of an increasingly clear choice. From the analysis of tools, dynamics and data, relevant opportunities for the organisation emerge, together with important conditions for their successful implementation. It is essential that the organization proceed without improvising, following, on one side, an

  9. The Gly2019Ser mutation in LRRK2 is not fully penetrant in familial Parkinson's disease: the GenePD study

    Corbett Alastair

    2008-11-01

    Full Text Available Abstract Background We report age-dependent penetrance estimates for leucine-rich repeat kinase 2 (LRRK2-related Parkinson's disease (PD in a large sample of familial PD. The most frequently seen LRRK2 mutation, Gly2019Ser (G2019S, is associated with approximately 5 to 6% of familial PD cases and 1 to 2% of idiopathic cases, making it the most common known genetic cause of PD. Studies of the penetrance of LRRK2 mutations have produced a wide range of estimates, possibly due to differences in study design and recruitment, including in particular differences between samples of familial PD versus sporadic PD. Methods A sample, including 903 affected and 58 unaffected members from 509 families ascertained for having two or more PD-affected members, 126 randomly ascertained PD patients and 197 controls, was screened for five different LRRK2 mutations. Penetrance was estimated in families of LRRK2 carriers with consideration of the inherent bias towards increased penetrance in a familial sample. Results Thirty-one out of 509 families with multiple cases of PD (6.1% were found to have 58 LRRK2 mutation carriers (6.4%. Twenty-nine of the 31 families had G2019S mutations while two had R1441C mutations. No mutations were identified among controls or unaffected relatives of PD cases. Nine PD-affected relatives of G2019S carriers did not carry the LRRK2 mutation themselves. At the maximum observed age range of 90 to 94 years, the unbiased estimated penetrance was 67% for G2019S families, compared with a baseline PD risk of 17% seen in the non-LRRK2-related PD families. Conclusion Lifetime penetrance of LRRK2 estimated in the unascertained relatives of multiplex PD families is greater than that reported in studies of sporadically ascertained LRRK2 cases, suggesting that inherited susceptibility factors may modify the penetrance of LRRK2 mutations. In addition, the presence of nine PD phenocopies in the LRRK2 families suggests that these susceptibility

  10. Polycystic kidney disease in the medaka (Oryzias latipes pc mutant caused by a mutation in the Gli-Similar3 (glis3 gene.

    Hisashi Hashimoto

    Full Text Available Polycystic kidney disease (PKD is a common hereditary disease in humans. Recent studies have shown an increasing number of ciliary genes that are involved in the pathogenesis of PKD. In this study, the Gli-similar3 (glis3 gene was identified as the causal gene of the medaka pc mutant, a model of PKD. In the pc mutant, a transposon was found to be inserted into the fourth intron of the pc/glis3 gene, causing aberrant splicing of the pc/glis3 mRNA and thus a putatively truncated protein with a defective zinc finger domain. pc/glis3 mRNA is expressed in the epithelial cells of the renal tubules and ducts of the pronephros and mesonephros, and also in the pancreas. Antisense oligonucleotide-mediated knockdown of pc/glis3 resulted in cyst formation in the pronephric tubules of medaka fry. Although three other glis family members, glis1a, glis1b and glis2, were found in the medaka genome, none were expressed in the embryonic or larval kidney. In the pc mutant, the urine flow rate in the pronephros was significantly reduced, which was considered to be a direct cause of renal cyst formation. The cilia on the surface of the renal tubular epithelium were significantly shorter in the pc mutant than in wild-type, suggesting that shortened cilia resulted in a decrease in driving force and, in turn, a reduction in urine flow rate. Most importantly, EGFP-tagged pc/glis3 protein localized in primary cilia as well as in the nucleus when expressed in mouse renal epithelial cells, indicating a strong connection between pc/glis3 and ciliary function. Unlike human patients with GLIS3 mutations, the medaka pc mutant shows none of the symptoms of a pancreatic phenotype, such as impaired insulin expression and/or diabetes, suggesting that the pc mutant may be suitable for use as a kidney-specific model for human GLIS3 patients.

  11. Assessment of the toll-like receptor 4 Asp299Gly, Thr399Ile and interleukin-8 -251 polymorphisms in the risk for the development of distal gastric cancer

    Maldonado-Garza Hector J

    2007-04-01

    Full Text Available Abstract Background The intensity of the inflammation induced by Helicobacter pylori colonization is associated with the development of distal gastric cancer (GC. The host response to H. pylori has been related to genetic polymorphisms that influence both innate and adaptive immune responses. Our aim was to investigate whether the presence of the TLR4 Asp299Gly, TLR4 Thr399Ile and IL-8-251 A/T polymorphisms had any influence in the development of distal GC in a Mexican population. Methods We studied 337 patients that were divided in two groups: 78 patients with histologically confirmed distal GC and 259 non-cancer controls. The presence of H. pylori in the control population was defined by positive results of at least two of four diagnostic tests: serology, histology, rapid urease test and culture. Human DNA was purified and genotyped for TLR4 Asp299Gly polymorphism by pyrosequencing, for TLR4 Thr399Ile by PCR-RFLP and for IL8-251 by the amplification refractory mutation system (ARMS-PCR. Results The non-cancer control group was found to be in Hardy-Weinberg equilibrium at the polymorphic loci studied (chi-square H-W = 0.58 for IL8-251, 0.42 for TLR4 Asp299Gly and 0.17 for TLR4 Thr399Ile. The frequencies of mutated alleles (homozygous plus heterozygous were compared between cases and controls. We found no significant difference for TLR4- Asp299Gly [the 7.7% of distal GC patients and 7.7 % non-cancer controls (p = 0.82] and for TLR4 Thr399Ile [the 1.3% of GC patients and the 5% of the control population (p = 0.2]. In contrast, for IL-8-251 A/T, 80.77% of the GC patients and 66.4% in the control group age and gender matched had at least one copy of mutated allele (OR = 2.12, 95% CI = 1.1–4.2 (p = 0.023. Conclusion This study showed that the IL8-251*A allele could be related to the development of distal gastric cancer in this Mexican population.

  12. Studies of the Gly482Ser polymorphism of the peroxisome proliferator-activated receptor gamma coactivator 1alpha (PGC-1alpha) gene in Danish subjects with the metabolic syndrome

    Ambye, Louise; Rasmussen, Susanne; Fenger, Mogens

    2005-01-01

    related to this syndrome. The variant was examined, using PCR-RFLP, in the DanMONICA cohort comprising a population-based sample of 2349 subjects. MS was defined using the National Cholesterol Education Program -- Adult Treatment Panel III (NCEP-ATPIII) criteria. The allelic frequency of the Ser482 allele...... and insulin secretion, 24-ambulatory blood pressure or left ventricular mass index. In conclusion, the Gly482Ser polymorphism of the PGC-1alpha gene is not associated with the metabolic syndrome, related quantitative traits or cardiac hypertrophy among Danish Caucasian subjects...

  13. Book review. Gli animali, l'uomo e l'ambiente. (a cura di Giorgio Battelli, Raffaella Baldelli, Fabio Ostanello, Santino Prosperi

    Manuel Graziani

    2013-12-01

    à di produzione e commercializzazione di alimenti di origine animale, alle emergenze di tipo sanitario, alle nuove istanze dei consumatori e degli amministratori pubblici. La prima parte "La storia e i concetti" si sofferma in particolare sull'evoluzione della SPV in Italia, sulle zoonosi, sul rapporto uomo-animale anche in ambito urbano con l'ultimo contributo relativo all'igiene urbana veterinaria. La seconda parte "Le attività" passa in rassegna i Servizi Veterinari pubblici del Paese, i NAS, gli Enti scientifici (tra cui IIZZSS e ISS fino a trattare attività quali la sicurezza alimentare, le emergenze non epidemiche, il benessere animale e la pet therapy. Nella terza e ultima parte "Le sfide" ci si interroga sul ruolo sociale della medicina veterinaria, sul concetto di medicina unica, sull'etica nella sperimentazione animale, sulla formazione, l'educazione e la comunicazione.

  14. Technetium-99m-labeled Arg-Gly-Asp-conjugated alpha-melanocyte stimulating hormone hybrid peptides for human melanoma imaging

    Yang Jianquan; Guo Haixun [College of Pharmacy, University of New Mexico, Albuquerque, NM 87131 (United States); Miao Yubin, E-mail: ymiao@salud.unm.ed [College of Pharmacy, University of New Mexico, Albuquerque, NM 87131 (United States); Cancer Research and Treatment Center, University of New Mexico, Albuquerque, NM 87131 (United States); Department of Dermatology, University of New Mexico, Albuquerque, NM 87131 (United States)

    2010-11-15

    Introduction: The purpose of this study was to examine whether {sup 99m}Tc-labeled Arg-Gly-Asp (RGD)-conjugated alpha-melanocyte stimulating hormone ({alpha}-MSH) hybrid peptide targeting both melanocortin-1 (MC1) and {alpha}{sub v{beta}3} integrin receptors was superior in melanoma targeting to {sup 99m}Tc-labeled {alpha}-MSH or RGD peptide targeting only the MC1 or {alpha}{sub v{beta}3} integrin receptor. Methods: RGD-Lys-(Arg{sup 11})CCMSH, RAD-Lys-(Arg{sup 11})CCMSH and RGD-Lys-(Arg{sup 11})CCMSHscramble were designed to target both MC1 and {alpha}{sub v{beta}3} integrin receptors, MC1 receptor only and {alpha}{sub v{beta}3} integrin receptor only, respectively. The MC1 or {alpha}{sub v{beta}3} integrin receptor binding affinities of three peptides were determined in M21 human melanoma cells. The melanoma targeting properties of {sup 99m}Tc-labeled RGD-Lys-(Arg{sup 11})CCMSH, RAD-Lys-(Arg{sup 11})CCMSH and RGD-Lys-(Arg{sup 11})CCMSHscramble were determined in M21 human melanoma-xenografted nude mice. Meanwhile, the melanoma uptake of {sup 99m}Tc-RGD-Lys-(Arg{sup 11})CCMSH was blocked with various non-radiolabeled peptides in M21 melanoma xenografts. Results: RGD-Lys-(Arg{sup 11})CCMSH displayed 2.0 and 403 nM binding affinities to both MC1 and {alpha}{sub v{beta}3} integrin receptors, whereas RAD-Lys-(Arg{sup 11})CCMSH or RGD-Lys-(Arg{sup 11})CCMSHscramble lost their {alpha}{sub v{beta}3} integrin receptor binding affinity by greater than 248-fold or MC1 receptor binding affinity by more than 100-fold, respectively. The melanoma uptake of {sup 99m}Tc-RGD-Lys-(Arg{sup 11})CCMSH was 2.49 and 2.24 times (P < .05) the melanoma uptakes of {sup 99m}Tc-RAD-Lys-(Arg{sup 11})CCMSH and {sup 99m}Tc-RGD-Lys-(Arg{sup 11})CCMSHscramble at 2 h post-injection, respectively. Either RGD or (Arg{sup 11})CCMSH peptide co-injection could block 42% and 57% of the tumor uptake of {sup 99m}Tc-RGD-Lys-(Arg{sup 11})CCMSH, whereas the coinjection of RGD+(Arg{sup 11})CCMSH peptide mixture

  15. Studies of the Gly482Ser polymorphism of the peroxisome proliferator-activated receptor gamma coactivator 1alpha (PGC-1alpha) gene in Danish subjects with the metabolic syndrome

    Ambye, L; Rasmussen, S; Fenger, Mogens

    2005-01-01

    The peroxisome proliferator-activated receptor gamma co-activator 1alpha (PGC-1alpha) is a novel transcriptional co-activator that holds an important role in lipid and glucose metabolism. PGC-1alpha is a candidate gene for the metabolic syndrome (MS) as well as type 2 diabetes. Recent studies...... related to this syndrome. The variant was examined, using PCR-RFLP, in the DanMONICA cohort comprising a population-based sample of 2349 subjects. MS was defined using the National Cholesterol Education Program -- Adult Treatment Panel III (NCEP-ATPIII) criteria. The allelic frequency of the Ser482 allele...... and insulin secretion, 24-ambulatory blood pressure or left ventricular mass index. In conclusion, the Gly482Ser polymorphism of the PGC-1alpha gene is not associated with the metabolic syndrome, related quantitative traits or cardiac hypertrophy among Danish Caucasian subjects...

  16. Modulation of Colorectal Cancer Risk by Polymorphisms in 51Gln/His, 64Ile/Val, and 148Asp/Glu of APEX Gene; 23Gly/Ala of XPA Gene; and 689Ser/Arg of ERCC4 Gene

    L. Dziki

    2017-01-01

    Full Text Available Polymorphisms in DNA repair genes may affect the activity of the BER (base excision repair and NER (nucleotide excision repair systems. Using DNA isolated from blood taken from patients (n=312 and a control group (n=320 with CRC, we have analyzed the polymorphisms of selected DNA repair genes and we have demonstrated that genotypes 51Gln/His and 148Asp/Glu of APEX gene and 23Gly/Ala of XPA gene may increase the risk of colorectal cancer. At the same time analyzing the gene-gene interactions, we suggest the thesis that the main factor to be considered when analyzing the impact of polymorphisms on the risk of malignant transformation should be intergenic interactions. Moreover, we are suggesting that some polymorphisms may have impact not only on the malignant transformation but also on the stage of the tumor.

  17. The novel RAF1 mutation p.(Gly361Ala) located outside the kinase domain of the CR3 region in two patients with Noonan syndrome, including one with a rare brain tumor.

    Harms, Frederike L; Alawi, Malik; Amor, David J; Tan, Tiong Y; Cuturilo, Goran; Lissewski, Christina; Brinkmann, Julia; Schanze, Denny; Kutsche, Kerstin; Zenker, Martin

    2018-02-01

    Noonan syndrome is characterized by typical craniofacial dysmorphism, postnatal growth retardation, congenital heart defect, and learning difficulties and belongs to the RASopathies, a group of neurodevelopmental disorders caused by germline mutations in genes encoding components of the RAS-MAPK pathway. Mutations in the RAF1 gene are associated with Noonan syndrome, with a high prevalence of hypertrophic cardiomyopathy (HCM). RAF1 mutations cluster in exons encoding the conserved region 2 (CR2), the kinase activation segment of the CR3 domain, and the C-terminus. We present two boys with Noonan syndrome and the identical de novo RAF1 missense variant c.1082G>C/p.(Gly361Ala) affecting the CR3, but located outside the kinase activation segment. The p.(Gly361Ala) mutation has been identified as a RAF1 allele conferring resistance to RAF inhibitors. This amino acid change favors a RAF1 conformation that allows for enhanced RAF dimerization and increased intrinsic kinase activity. Both patients with Noonan syndrome showed typical craniofacial dysmorphism, macrocephaly, and short stature. One individual developed HCM and was diagnosed with a disseminated oligodendroglial-like leptomeningeal tumor (DOLT) of childhood at the age of 9 years. While there is a well-established association of NS with malignant tumors, especially childhood hemato-oncological diseases, brain tumors have rarely been reported in Noonan syndrome. Our data demonstrate that mutation scanning of the entire coding region of genes associated with Noonan syndrome is mandatory not to miss rare variants located outside the known mutational hotspots. © 2017 Wiley Periodicals, Inc.

  18. A cyclic carbo-isosteric penta-depsipeptide: cyclo(Phe1–d-Ala2–Gly3–Phe4–APO5

    Stéphanie M. Guéret

    2015-01-01

    Full Text Available The title compound, cyclo(Phe1–d-Ala2–Gly3–Phe4–APO5, C26H32N4O5, is the minor diastereoisomer of a cyclic penta-peptidomimetic analogue containing a novel 2-aminopropyl lactone (APO motif, which displays the same number of atoms as the native amino acid glycine and has a methyl group in place of the carbonyl O atom. The crystal structure presented here allows the analysis of the secondary structure of this unprecedented cyclic carbo-isosteric depsipeptide. The conformation of the central ring is stabilized by an intramolecular N—H...O hydrogen bond between the carbonyl O atom of the first residue (Phe1 and the amide group H atom of the fourth residue (Phe4. Based on the previously reported hydrogen bond and on the values of the torsion angles ϕ and ψ, the loop formed by the first, second, third and fourth residues (Phe1, d-Ala2, Gly3 and Phe4 can be classified as a type II′ β-turn. The loop around the new peptidomimetic motif, on the other hand, resembles an open γ-turn containing a weak N—H...O hydrogen bond between the carbonyl group O atom of the fourth residue (Phe4 and the amide unit H atom of the first residue (Phe1. In the crystal, the peptidomimetic molecules are arranged in chains along the b-axis direction. Within such a chain, the molecules of the structure are linked via N—H...O hydrogen bonds between the amide group H atom of the secondary residue (d-Ala2 and the carboxy unit O atom of the fourth residue (Phe4 in a neighboring molecule. The newly formed methyl stereocentre of the APO peptidomimetic motif (APO5 was obtained as the minor diastereoisomer in a ring-closing reductive amination reaction and adopts an R configuration.

  19. Supplementation of antipsychotic treatment with sarcosine – GlyT1 inhibitor – causes changes of glutamatergic (1)NMR spectroscopy parameters in the left hippocampus in patients with stable schizophrenia.

    Strzelecki, Dominik; Podgórski, Michał; Kałużyńska, Olga; Gawlik-Kotelnicka, Oliwia; Stefańczyk, Ludomir; Kotlicka-Antczak, Magdalena; Gmitrowicz, Agnieszka; Grzelak, Piotr

    2015-10-08

    Glutamatergic system, the main stimulating system of the brain, plays an important role in the pathogenesis of schizophrenia. Hippocampus, a structure crucial for memory and cognitive functions and rich in glutamatergic neurons, is a natural object of interest in studies on psychoses. Sarcosine, a glycine transporter (GlyT-1) inhibitor influences the function of NMDA receptor and glutamate-dependent transmission. The aim of the study was to assess the effects of sarcosine on metabolism parameters in the left hippocampus in patients with schizophrenia. Assessments were performed using proton nuclear magnetic resonance ((1)H NMR) spectroscopy (1.5T). Fifty patients diagnosed with schizophrenia (DSM-IV-TR), with dominant negative symptoms, in stable clinical condition and stable antipsychotics doses were treated either with sarcosine (n=25) or placebo (n=25). Spectroscopic parameters were evaluated within groups and between two groups before and after 6-month intervention. All patients were also assessed with the Positive and Negative Syndrome Scale (PANSS). In the sarcosine group, after 6-month treatment, we found significant decrease in hippocampal Glx/Cr (Glx-complex of glutamate, glutamine and GABA, Cr-creatine) and Glx/Cho (Cho-choline), while N-acetylaspartate (NAA), myo-inositol (mI), Cr and Cho parameters remained stable along the study and also did not differ significantly between both groups. This is the first study showing that a pharmacological intervention in schizophrenia, particularly augmentation of the antypsychotic treatment with sarcosine, may reverse the pathological increase in glutamatergic transmission in the hippocampus. The results confirm involvement of glutamatergic system in the pathogenesis of schizophrenia and demonstrate beneficial effects of GlyT-1 inhibitor on the metabolism in the hippocampus and symptoms of schizophrenia. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  20. Dimer of the peptide cycle (Ar-Gly-Asp-D-Phe-Lys) radiolabeled with {sup 99m}Tc for the integrin s over-expression image: formulation, biokinetics and dosimetry; Dimero del peptido ciclo(Arg-Gly-Asp-D-Phe-Lys) radiomarcado con {sup 99m}Tc para la imagen de sobre-expresion de integrinas: formulacion, biocinetica y dosimetria

    Ortiz A, Z.

    2013-07-01

    In breast cancer, α(v)β(3) and/or α(v)β(5) integrin s are over-expressed in both endothelial and tumour cells. Radiolabeled peptides based on the RGD (Arg-Gly-Asp) sequence are radiopharmaceuticals with high affinity and selectivity for those integrin s. The RGD-dimer peptide (E-[c(RGDfK)]{sub 2}) radiolabeled with {sup 99m}Tc has been reported as a radiopharmaceutical with 10-fold higher affinity for the α(v)β(3) integrin as compared to the RGD-monomer. EDDA (Ethylenediamine-N,N-diacetic acid) is a hydrophilic molecule that may favours renal excretion when used as coligand in the {sup 99m}Tc labelling of HYNIC-peptides and can easily be formulated in a lyophilized kit. Aim: Establish a biokinetic model for {sup 99m}Tc-EDDA/HYNIC-E-[c(RGDfK)]{sub 2} prepared from lyophilized kits and evaluate the dosimetry as breast cancer imaging agent. Methods: {sup 99m}Tc labelling was performed by addition of sodium pertechnetate solution and 0.2 M phosphate buffer ph 7.0 to a lyophilized formulation containing E-[c(RGDfK)]{sub 2}, EDDA, tricine, mannitol and stannous chloride. Radiochemical purity was evaluated by reversed phase HPLC and ITLC-SG analyses. Stability studies in human serum were carried out by size-exclusion HPLC. In-vitro cell uptake was tested using breast cancer cells (MCF7, T47D and MDA-MB-231) with blocked and non-blocked receptors. Biodistribution and tumour uptake were determined in MCF7 tumour-bearing nude mice with blocked and non-blocked receptors, and images were obtained using a micro-SPECT/CT. Whole-body images from seven healthy women were acquired at 1, 3, 6 and 24 h after {sup 99m}Tc-EDDA/HYNIC-E-[c(RGDfK)]{sub 2} administration obtained with radiochemical purities of >94 %. Regions of interest (ROIs) were drawn around source organs on each time frame. Each ROI was converted to activity using the conjugate view counting method. The image sequence was used to extrapolate {sup 99m}Tc-EDDA/HYNIC-E-[c(RGDfK)]{sub 2} time-activity curves in each

  1. A novel c.1037C > G (p.Ala346Gly) mutation in TP63 as cause of the ectrodactyly-ectodermal dysplasia and cleft lip/palate (EEC) syndrome

    Alves, Leandro Ucela; Pardono, Eliete; Otto, Paulo A.; Mingroni Netto, Regina Célia

    2015-01-01

    Ectrodactyly – ectodermal dysplasia and cleft lip/palate (EEC) syndrome (OMIM 604292) is a rare disorder determined by mutations in the TP63 gene. Most cases of EEC syndrome are associated to mutations in the DNA binding domain (DBD) region of the p63 protein. Here we report on a three-generation Brazilian family with three individuals (mother, son and grandfather) affected by EEC syndrome, determined by a novel mutation c.1037C > G (p.Ala346Gly). The disorder in this family exhibits a broad spectrum of phenotypes: two individuals were personally examined, one presenting the complete constellation of EEC syndrome manifestations and the other presenting an intermediate phenotype; the third affected, a deceased individual not examined personally and referred to by his daughter, exhibited only the split-hand/foot malformation (SHFM). Our findings contribute to elucidate the complex phenotype-genotype correlations in EEC syndrome and other related TP63-mutation syndromes. The possibility of the mutation c.1037C > G being related both to acro-dermato-ungual-lacrimal-tooth (ADULT) syndrome and SHFM is also raised by the findings here reported. PMID:25983622

  2. Physicochemical Properties of Glycine-Based Ionic Liquid [QuatGly-OEt][EtOSO3] (2-Ethoxy-1-ethyl-1,1-dimethyl-2-oxoethanaminium ethyl sulfate and Its Binary Mixtures with Poly(ethylene glycol (Mw = 200 at Various Temperatures

    Chung-Wen Kuo

    2011-12-01

    Full Text Available This work includes specific basic characterization of synthesized glycine-based Ionic Liquid (IL [QuatGly-OEt][EtOSO3] by NMR, elementary analysis and water content. Thermophysical properties such as density, ρ, viscosity, η, refractive index, n, and conductivity, κ, for the binary mixture of [QuatGly-OEt][EtOSO3] with poly(ethylene glycol (PEG [Mw = 200] are measured over the whole composition range. The temperature dependence of density and dynamic viscosity for neat [QuatGly-OEt][EtOSO3] and its binary mixture can be described by an empirical polynomial equation and by the Vogel-Tammann-Fucher (VTF equation, respectively. The thermal expansion coefficient of the ILs is ascertained using the experimental density results, and the excess volume expansivity is evaluated. The negative values of excess molar volume for the mixture indicate the ion-dipole interactions and packing between IL and PEG oligomer. The results of binary excess property (VmE and deviations (Δη, ∆xn, ∆Фn, ∆xR, and ∆ФR are discussed in terms of molecular interactions and molecular structures in the binary mixture.

  3. Physicochemical Properties of Glycine-Based Ionic Liquid [QuatGly-OEt][EtOSO3] (2-Ethoxy-1-ethyl-1,1-dimethyl-2-oxoethanaminium ethyl sulfate) and Its Binary Mixtures with Poly(ethylene glycol) (Mw = 200) at Various Temperatures

    Wu, Tzi-Yi; Chen, Bor-Kuan; Hao, Lin; Lin, Yuan-Chung; Wang, H. Paul; Kuo, Chung-Wen; Sun, I-Wen

    2011-01-01

    This work includes specific basic characterization of synthesized glycine-based Ionic Liquid (IL) [QuatGly-OEt][EtOSO3] by NMR, elementary analysis and water content. Thermophysical properties such as density, ρ, viscosity, η, refractive index, n, and conductivity, κ, for the binary mixture of [QuatGly-OEt][EtOSO3] with poly(ethylene glycol) (PEG) [Mw = 200] are measured over the whole composition range. The temperature dependence of density and dynamic viscosity for neat [QuatGly-OEt][EtOSO3] and its binary mixture can be described by an empirical polynomial equation and by the Vogel-Tammann-Fucher (VTF) equation, respectively. The thermal expansion coefficient of the ILs is ascertained using the experimental density results, and the excess volume expansivity is evaluated. The negative values of excess molar volume for the mixture indicate the ion-dipole interactions and packing between IL and PEG oligomer. The results of binary excess property (VmE ) and deviations (Δη, Δxn, ΔΨn, ΔxR, and ΔΨR) are discussed in terms of molecular interactions and molecular structures in the binary mixture. PMID:22272102

  4. Orchestration of Structural, Stereoelectronic, and Hydrogen-Bonding Effects in Stabilizing Triplexes from Engineered Chimeric Collagen Peptides (Pro(X)-Pro(Y)-Gly)6 Incorporating 4(R/S)-Aminoproline.

    Umashankara, Muddegowda; Sonar, Mahesh V; Bansode, Nitin D; Ganesh, Krishna N

    2015-09-04

    Collagens are an important family of structural proteins found in the extracellular matrix with triple helix as the characteristic structural motif. The collagen triplex is made of three left-handed polyproline II (PPII) helices with each PPII strand consisting of repetitive units of the tripeptide motif X-Y-Gly, where the amino acids X and Y are most commonly proline (Pro) and 4R-hydroxyproline (Hyp), respectively. A C4-endo pucker at X-site and C4-exo pucker at Y-site have been proposed to be the key for formation of triplex, and the nature of pucker is dependent on both the electronegativity and stereochemistry of the substituent. The present manuscript describes a new class of collagen analogues-chimeric cationic collagens-wherein both X- and Y-sites in collagen triad are simultaneously substituted by a combination of 4(R/S)-(OH/NH2/NH3(+)/NHCHO)-prolyl units and triplex stabilities measured at different pHs and in EG:H2O. Based on the results a model has been proposed with the premise that any factors which specifically favor the ring puckers of C4-endo at X-site and C4-exo at Y-site stabilize the PPII conformation and hence the derived triplexes. The pH-dependent triplex stability uniquely observed with ionizable 4-amino substituent on proline enables one to define the critical combination of factors C4-(exo/endo), intraresidue H-bonding, stereoelectronic (R/S) and n → π* interactions in dictating the triplex strength. The ionizable NH2 substituent at C4 in R/S configuration is thus a versatile probe for delineating the triplex stabilizing factors and the results have potential for designing of collagen analogues with customized properties for material and biological applications.

  5. Biological evaluation of 177Lu-labeled DOTA-Ala(SO3H)-Aminooctanoyl-Gln-Trp-Ala-Val-N methyl Gly-His-Statine-Leu-NH2 for gastrin-releasing peptide receptor-positive prostate tumor targeting

    Lim, Jae Cheong; Cho, Eun Ha; Kim, Jin Joo; Choi, Sang Mu; Lee, So young; Nam, Sung Soo; Park, Ul Jae; Park, Soo Hyun

    2015-01-01

    Bombesin binds with selectivity and high affinity to a Gastrin-releasing peptide receptor (GRPR), which is highly overexpressed in prostate cancer cells. The present study describes the in vitro and in vivo biological characteristics of DOTA-Ala(SO 3 H)-Aminooctanoyl-Gln-Trp-Ala-Val-N methyl Gly-His-Statine-Leu-NH 2 (DOTA-sBBNA), an antagonist analogue of bombesin peptide for the targeting of GRPR. DOTA-sBBNA was synthesized and labeled with 177 Lu as previously published. A saturation assay on PC-3 human prostate cancer cells revealed that the Kd value of the radiolabeled peptide was 1.88 nM with a maximum binding capacity (Bmax) of 289.3 fmol/10 6 cells. The radio-peptide slowly internalized, and 24.4 ± 0.5% of the total binding was internalized in 4 hr. Biodistribution studies were conducted in healthy and PC-3 xenografted balb/c mice, which showed high uptake and retention of tumor-associated radioactivity in PC-3 xenografted mice. The tumor-to-blood ratio was 126.02 ± 9.36 at 1.5 hr p.i., and was increased to 216.33 ± 61.58 at 24 hr p.i., which means that the radiolabeled peptide was highly accumulated in a tumor and rapidly cleared from the blood pool. The GRPR is also over-expressed in Korean prostate cancer patients. These results suggest that this 177 Lu-labeled peptide has promising characteristics for application in nuclear medicine, namely for the diagnosis and treatment of GRPR over-expressing prostate tumors

  6. L’Inquisizione, gli indizi, le prove

    Guido Dall’Olio

    2012-11-01

    Full Text Available This essay focuses on some characteristics of the inquisitorial trial, comparing it with the accusatory procedure. The inquisitorial trial, which resulted in almost inevitable torture of the accused, is certainly in our eyes an injustice and a violation of the legal rights that are essential for us in a proper procedure. However, the care with which the evidence was produced and examined by the institutions that adopted the inquisitorial procedure deserves to be emphasized, especially for the consequences it had in certain types of imaginary crime such as witchcraft.

  7. Dimer of the peptide cycle (Ar-Gly-Asp-D-Phe-Lys) radiolabeled with 99mTc for the integrin s over-expression image: formulation, biokinetics and dosimetry

    Ortiz A, Z.

    2013-01-01

    In breast cancer, α(v)β(3) and/or α(v)β(5) integrin s are over-expressed in both endothelial and tumour cells. Radiolabeled peptides based on the RGD (Arg-Gly-Asp) sequence are radiopharmaceuticals with high affinity and selectivity for those integrin s. The RGD-dimer peptide (E-[c(RGDfK)] 2 ) radiolabeled with 99m Tc has been reported as a radiopharmaceutical with 10-fold higher affinity for the α(v)β(3) integrin as compared to the RGD-monomer. EDDA (Ethylenediamine-N,N-diacetic acid) is a hydrophilic molecule that may favours renal excretion when used as coligand in the 99m Tc labelling of HYNIC-peptides and can easily be formulated in a lyophilized kit. Aim: Establish a biokinetic model for 99m Tc-EDDA/HYNIC-E-[c(RGDfK)] 2 prepared from lyophilized kits and evaluate the dosimetry as breast cancer imaging agent. Methods: 99m Tc labelling was performed by addition of sodium pertechnetate solution and 0.2 M phosphate buffer ph 7.0 to a lyophilized formulation containing E-[c(RGDfK)] 2 , EDDA, tricine, mannitol and stannous chloride. Radiochemical purity was evaluated by reversed phase HPLC and ITLC-SG analyses. Stability studies in human serum were carried out by size-exclusion HPLC. In-vitro cell uptake was tested using breast cancer cells (MCF7, T47D and MDA-MB-231) with blocked and non-blocked receptors. Biodistribution and tumour uptake were determined in MCF7 tumour-bearing nude mice with blocked and non-blocked receptors, and images were obtained using a micro-SPECT/CT. Whole-body images from seven healthy women were acquired at 1, 3, 6 and 24 h after 99m Tc-EDDA/HYNIC-E-[c(RGDfK)] 2 administration obtained with radiochemical purities of >94 %. Regions of interest (ROIs) were drawn around source organs on each time frame. Each ROI was converted to activity using the conjugate view counting method. The image sequence was used to extrapolate 99m Tc-EDDA/HYNIC-E-[c(RGDfK)] 2 time-activity curves in each organ in order to adjust the biokinetic model and to

  8. Il concetto di sicurezza e la sua percezione. Gli studenti universitari si interrogano/Le concept de sécurité et sa perception. Les étudiants universitaires s'interrogent/Security concept and its perception. Questionnaire for University students

    Anastasia Moschi

    2013-12-01

    Full Text Available Nell’articolo vengono evidenziati i risultati emersi da una ricerca, concernente la percezione della sicurezza, condotta tra gli studenti universitari nell’anno 2012. Obiettivo dell’indagine era quello di cogliere se, e come, l’appartenenza a corsi di laurea differenti potesse incidere sul modo di trattare e considerare la “questione sicurezza”. Sono emerse, effettivamente, differenze rilevanti nell’approccio al tema in questione declinato nelle sue componenti principali (paura del crimine, vittimizzazione reale e potenziale, interventi auspicabili, preoccupazioni, ruoli istituzionali e dei cittadini. Al contempo però appare evidente anche la necessità che tutti i percorsi universitari e tutti gli attori sociali, anche se da prospettive differenti, si confrontino con il fenomeno-sicurezza per sviluppare una collaborazione tra professionalità diverse che possa condurre verso una cultura della sicurezza. Cet article récapitule les principaux résultats obtenus par une enquête sur la perception de la sécurité menée auprès d’étudiants universitaires en 2012. La présente étude avait pour but d’examiner si le fait d’être inscrit à des cours differents et à différents niveaux pourrait influencer les ideés des étudiants à propos des questions en matière de sécurité. En fait, beaucoup de différences significatives liées à ce sujet ont été remarquées, tels que la peur de la criminalité, le risque réel et potentiel d’être victime de crimes, les mesures d’intervention à envisager, les rôles des institutions et des citoyens. En même temps, l’enquête a souligné la necessité pour les étudiants inscrits à tous les cours et pour tous les acteurs sociaux de se rapprocher au phénomène de la sécurité dans le but de développer la coopération entre différents professionnels en vue d’élaborer une culture commune de la sécurité. This article shows the results of a survey about the perception

  9. Hb Belluno [β111(G13)Val→Gly;β133(H11)Val→Val (HBB: c.335T > G;402G > C)]: Incidental Detection of a New Clinically Silent β Chain Variant During Hb A1c Determination by High Performance Liquid Chromatography.

    Pianezze, Graziano; Toniolo, Manuele; Taddei Masieri, Marina; Dolcini, Bernardetta; Ravani, Anna

    2016-06-01

    A previously unreported β chain variant, Hb Belluno [β111(G13)Val→Gly;β133(H11)Val→Val (HBB: c.335T > G;402G > C)], was incidentally discovered in a woman suffering from diabetes, during glycated hemoglobin (Hb A1c) assay. Its presence was suspected because of a small abnormal peak with a retention time just shorter than that of normal Hb A1c. Standard high performance liquid chromatography (HPLC), capillary zone electrophoresis (CZE) and agarose gel electrophoresis did not allow to separate the variant from Hb A. The reversed phase HPLC of globin chains showed the presence of a heterozygous β-globin variant amounting to approximately 43.5% of the total β chains. Later, this variant was found in five other members of the same family and DNA sequencing analysis confirmed a β-globin gene mutation. The variant is clinically silent in all patients and showed a slight instability with both heat and isopropanol tests. The other three mutations at this locus also affect stability. Hemoglobin (Hb) variants may invalidate the results of Hb A1c analysis and could result in mismanagement of diabetes. A comment alerting the requesting clinician to the presence of the Hb variant must be appended to the Hb A1c result. Additionally, many Hb variants can be chromatographically and/or electrophoretically silent. Therefore, when the clinician suspects a variant Hb, it is not sufficient to get a negative response from an HPLC screening test to rule it out. A dialogue with the pathologist is essential, involving exchange of information and sharing a diagnostic work-up including surveys to assess Hb stability and oxygen affinity, as much as DNA sequencing.

  10. Note sulla concezione del post mortem presso gli Ittiti

    Vigo, Matteo; Bellucci, Benedetta

    significato escatologico della morte; 2) la definizione dello status ultraterreno dei defunti (i.e. il diverso trattamento del defunto, ad esempio dal punto di vista cultuale, sulla base della sua condizione sociale); 3) il delineamento dei caratteri e delle funzioni delle divinità ultra-terrene; 4) la......Oggetto d’analisi di questo paper è la concezione della sfera del post-mortem presso la civiltà degli Ittiti. Lo studio delle tematiche che riguardano la concezione dell’“aldilà” nel mondo ittita, sia in senso fisico che metafisico, riveste una larga importanza nella letteratura ittitologica ed è...... stato, pertanto, diffusamente trattato. Nel presente contributo si è scelto quindi di approfondire solo alcuni aspetti della concezione dell’oltretomba, inteso come dimensione metafisica in cui il corpo del defunto giace, staziona o transita dopo la morte. I temi approfonditi saranno quindi: 1) il...

  11. Una complessa chiarezza: gli ipertesti di Primo Levi

    Tommaso Pepe

    2016-11-01

    Full Text Available Scrittore poliedrico e versatile, Primo Levi è stato anche un costruttore di estesi e articolati ipertesti: strutture macrotestuali fondate su legami e interrelazioni tese fra opere cronologicamente e tipologicamente differenziate. Un esempio straordinario, ma non certo isolato, di questa scrittura di secondo grado, organizzata in complessi palinsesti testuali, è quello che già Marco Belpoliti ha definito nei termini di «macrotesto del Lager»: un reticolo intra- e intertestuale che, a partire da Se questo è un uomo, si è sviluppato con un andamento rizomatico attraverso diverse stratificazioni scrittorie e genealogiche, sino a I sommersi e i salvati. Il «rizoma» tuttavia è semplicemente una delle varie macrostrutture individuabili nell’opera di Levi. Attraverso un confronto contrastivo con l’opera di Calvino, il saggio si propone di indagare alcune delle caratteristiche salienti di questa scrittura ipertestuale, mostrando come l’intensa sperimentazione scrittoria perseguita da Levi sia da ricollegarsi a una parallela svolta epistemologica, data dall’insorgere di quella che Edgar Morin ha definito come «sfida della complessità».Primo Levi can be considered a versatile and polyhedral writer, capable of building long and articulated hypertexts, i.e., macrotextual structures based on the links and interrelations among works which differ both from a chronological and typological point of view. An extraordinary, but certainly not isolated, example of this writing, organized according to complex textual palimpsests, is the one that Marco Belpoliti defined as the «macrotext of the Lager»: it is an intra- and intertextual network that, starting from If This is a Man, developed according to a rhizomatic structure through his different works and genealogical stratifications, ending with The Drowned and the Saved. The «rhizome», however, is only one of the different macrostructures which may be singled out in Levi’s work. By developing a contrastive comparison with the work of Calvino, this essay aims to investigate some of the salient features of such hypertextual writing, thus showing how the marked textual experimentation pursued by Levi has to be linked to a parallel epistemological turn, given by the emergence of what Edgar Morin defined as the «challenge of complexity». PAROLE CHIAVEPrimo Levi; Italo Calvino; ipertesto; rizoma; complessità. 

  12. Gli amministratori indipendenti nelle società quotate

    Della Vedova, Ilaria

    2014-01-01

    This thesis is focused on independent directors and on the role they can play in companies characterized by concentrated ownership. Particular attention is dedicated to independent directors in listed companies and in banks. The survey starts analyzing the rise and development of independent directors in common law systems – that is, in the systems where they were first born – in order to highlight the role these directors have and the interests of shareholders they are asked to protect as...

  13. Purim e gli albori del teatro ebraico in Italia

    Maria Luisa Mayer Modena

    2011-06-01

    Full Text Available The Jewish theatre in Hebrew has its very first beginning in Italy with the "Wedding Comedy" by Leone de Sommi (Mantua, floruit 1550, whose inspiration owes much to the Purim tradition. Jewish theatre, especially the Judaeo-Italian one, continues to flourish until XX century,  always maintaining a strong connection with the Purim celebration. This article will deal with the development of the Jewish theatre - both in Hebrew and in Jewish language -, with  particular emphasis on its relationship with the Purim. In examining the main phases of such  a relevant phenomenon we will rely on the documents currently known, with particular reference  to the Judaeo Italian corpus; one of the first objectives of this work will be to open new perspectives for further research in the field of Jewish dramatic literature.

  14. 3_gly_"&o {ft_y[:! *ogl9_ achieve

    until a terminal day-length of 14 hours at 2O weeks of age was achieved. The control treatment was a ..... this experiment than in the previous experiment (Mac- lachlan et aL, 1977) is in close agreement with ... Grouping of eggs accortling to mass os a percentage of the totol laid, to 28 weeks of age and production per pullet ...

  15. Gli effetti della rabbia nei giochi di coordinamento

    Pietroni, D.; van Kleef, G.A.; Steinel, W.; Alparone, F.R.

    2008-01-01

    The effects of anger in coordination games In coordination games agents' strategies often differ from normative models' prescriptions. Most games are characterized by divergence of interests among agents. Since divergences may elicit strong emotional reactions which may affect agents' coordination

  16. Gli strumenti di contrasto all'economia mafiosa

    Simona Romanò

    2016-12-01

    Full Text Available Il contributo analizza i risultati di una ricerca empirica sulle misure di prevenzione patrimoniale applicate dal Tribunale di Milano negli ultimi quindici anni (2000-2015 nei confronti delle persone indiziate di appartenere a organizzazioni mafiose. Accanto agli aspetti quantitativi della ricerca, viene approfondito il versante qualitativo delle misure di contrasto alla ricchezza mafiosa attraverso lo studio dei casi più significativi. Parole chiave: misure di prevenzione patrimoniale; confisca; amministrazione giudiziaria; economia mafiosa The essay analyzes the results of an empirical research on preventive measures imposed by the Court of Milan in the last fifteen years (2000-2015 on persons suspected of being members of Mafia-type organizations. Alongside quantitative aspects, it has been carried out a qualitative analysis of confiscation measures against organized crime, through a study of the most significant cases. Keywords: preventive measures; confiscation; organized crime

  17. Personaggio e Rivoluzione. Robespierre, Danton e gli altri

    Emanuela Piga

    2016-05-01

    Full Text Available This study aims to enquire the account of the French Revolution through the form of the novel and the building of characters, both invented or historically existed and reconstructed through the lens of the fiction. Starting from Victor Hugo’s nineteen century novel Quatre-vingt-treize (1874, the goal is to intertwine the analysis of the historiographical perspective inherent to the storytelling with the construction of different characters, core instrument of the ”melodramatic imagination” (P. Brooks. In this direction, the goal is also to reflect on the figuralization of moral conflicts which characterize the age of French Revolution and of the Jacobin ideology during the Terror.

  18. Mutation of the conserved Gly83 and Gly94 in Escherichia coli elongation factor Tu. Indication of structural pivots

    Kjaersgård, I V; Knudsen, Charlotte Rohde; Wiborg, O

    1995-01-01

    Elongation factor Tu from Escherichia coli cycles between an active conformation where GTP is bound, and an inactive conformation where GDP is bound. Between the two conformations, elongation factor Tu undergoes major structural changes. The aim of this work has been to reveal the role of two ver...... is an important pivot point in elongation factor-Tu. Udgivelsesdato: 1995-Feb-15...

  19. A single center, pilot, double-blinded, randomized, comparative, prospective clinical study to evaluate improvements in the structure and function of facial skin with tazarotene 0.1% cream alone and in combination with GliSODin® Skin Nutrients Advanced A

    Goldberg LD

    2014-05-01

    Full Text Available Lawrence D Goldberg, Corina CryslerShaft Medical San Diego, San Diego, CA, USABackground: Superoxide dismutase (SOD reduces the reactive oxygen species formation associated with oxidative stress. An imbalance between free radicals and antioxidants can lead to accelerated aging. GliSODin® Skin Nutrients Advanced Anti-Aging Formula (GAAF is an SOD-containing dietary nutricosmetic formulated with other nutraceuticals that promote improvements in the structure and function of the skin, including hydration, elasticity, structural integrity, and photoaging caused by oxidative stress. Tazarotene cream 0.1% (TAZ is a United States Food and Drug Administration-approved drug indicated for use in the mitigation of facial fine wrinkling, facial mottled hyper- and hypopigmentation, and benign facial lentigines when taken in conjunction with a comprehensive skin care and sun avoidance program.Objective: To determine if the antioxidant, anti-aging, hydrating and skin-rejuvenating properties of GAAF complement the retinoic actions of TAZ to improve the structure and function of facial skin.Method: A 90-day comparative study of ten subjects with facial photodamage; daily topical application of TAZ was used in combination with three capsules of GAAF (780 mg each or placebo orally, with food, per the randomization allocation.Results: After 90 days of treatment, TAZ alone and in combination with GAAF improved fine wrinkles (↓1.2 versus 2.0, mottled hyperpigmentation (↓2.2 versus 2.8 and overall photodamage (↓1.0 versus 1.8, as well as patient-reported response to treatment (↓2.0 versus 1.6. At week 12, TAZ/GAAF combination treatment (Group A versus TAZ treatment alone (Group C was of significant clinical benefit, with respect to fine wrinkling (14.7%/41.7%, overall photodamage (15.6%/53.0%, skin moisture (19.1%/103.2%, skin elasticity (12.8%/87.7%, and response to treatment (8.8%/21.4%.Conclusion: The study suggests GAAF in combination with TAZ is safe and

  20. Tc-99m Glu-Cys-Gly-His-Gly-Lys (ECG-HGK), a novel Tc-99m labeled hexapeptide for molecular tumor imaging.

    Kim, Dae-Weung; Kim, Myoung Hyoun; Kim, Chang Guhn

    2016-03-01

    Domain 5 of kinin-free high molecular weight kininogen inhibits the adhesion of many tumor cell lines, and it has been reported that the histidine-glycine-lysine (HGK)-rich region might be responsible for inhibition of cell adhesion. The authors developed HGK-containing hexapeptide, glutamic acid-cysteine-glycine (ECG)-HGK, and evaluated the utility of Tc-99m ECG-HGK for tumor imaging. Hexapeptide, ECG-HGK was synthesized using Fmoc solid-phase peptide synthesis. Radiolabeling efficiency was evaluated. The uptake of Tc-99m ECG-HGK within HT-1080 cells was evaluated in vitro. In HT-1080 tumor-bearing mice, gamma imaging and biodistribution studies were performed. The complexes Tc-99m ECG-HGK was prepared in high yield. The uptake of Tc-99m ECG-HGK within the HT-1080 tumor cells had been demonstrated by in vitro studies. The gamma camera imaging in the murine model showed that Tc-99m ECG-HGK was accumulated substantially in the HT-1080 tumor (tumor-to-muscle ratio = 5.7 ± 1.4 at 4 h), and the tumoral uptake was blocked by the co-injection of excess HGK (tumor-to-muscle ratio = 2.8 ± 0.6 at 4 h). In the present study, Tc-99m ECG-HGK was developed as a new tumor imaging agents. Our in vitro and in vivo studies revealed specific function of Tc-99m ECG-HGK for tumor imaging. Copyright © 2016 John Wiley & Sons, Ltd.

  1. Soybean (Glycine max) allergy in Europe: Gly m 5 (beta-conglycinin) and Gly m 6 (glycinin) are potential diagnostic markers for severe allergic reactions to soy

    Holzhauser, Thomas; Wackermann, Olga; Ballmer-Weber, Barbara K

    2008-01-01

    BACKGROUND: Soybean is considered an important allergenic food, but published data on soybean allergens are controversial. OBJECTIVE: We sought to identify relevant soybean allergens and correlate the IgE-binding pattern to clinical characteristics in European patients with confirmed soy allergy....

  2. GLI ERRORI DI ITALIANO L1 ED L2: INTERFERENZA E APPRENDIMENTO

    Rosaria Solarino

    2011-02-01

    Full Text Available Si può oggi affrontare il tema degli errori di italiano da una prospettiva che possa giovare contemporaneamente a docenti di italiano L1 ed L2? Noi pensiamo di sì: la ricerca glottodidattica sembra aver ormai apprestato un terreno comune alle due situazioni di apprendimento, sgombrando il campo da vecchi pregiudizi e distinzioni che appaiono ormai superate. Attraverso la contrapposizione di concetti quali “lingua parlata/lingua scritta”,  “errori di lingua / errori di linguaggio”, “apprendimento spontaneo/apprendimento guidato”, “italiano L1/italiano L2”, “errori di apprendimento/errori di interferenza, si indicano diversi criteri per la interpretazione degli errori e la loro valutazione in relazione alle cause, alle situazioni comunicative, ai contesti o allo stadio di evoluzione dell’apprendimento della lingua.     Errors in italian L1 and L2: interference and learning   Can errors in Italian be approached in a way that benefits both L1 and L2 Italian teachers? We believe so: glottodidactic research seems to have prepared a common terrain for these two learning situations, clearing the field of old prejudices and obsolete distinctions.  Through the juxtaposition of concepts like “spoken language/written language”, “language errors/speech errors”, “spontaneous learning/guided learning”, “L1 Italian/L2 Italian”, “learning errors/interference errors”, different criteria for interpreting errors and evaluating them in relation to their causes, to communicative situations, to contexts and the developmental state in learning a language are singled out.

  3. Osservazioni cliniche ed anatomiche nella storia della neuropsicologia del linguaggio: gli studi pre-Broca

    Claudio Luzzatti

    2013-08-01

    Full Text Available Clinical and Anatomical Observations in the History of Neurolinguistics: Studies Predating Paul Broca – Classical neurological and neuropsychological handbooks usually cite Paul Broca’s communication on the frontal lobes as the seat of articulated speech (1861 as the starting point in the history of the study of aphasia as. In their seminal study, Benton and Joynt (1960 have collected numerous observations of aphasic patients from the Greek, Latin Medieval and Renaissance medical tradition and from the eighteenth-century Enlightenment literature. In none of the reported cases, however, were the crucial elements of aphasia, that is, is, on the one hand the distinction between language deficits, conceptual damage and speech disorders and, on the other hand, the association with lesions of the left brain hemisphere – clearly identified. The first part of this article describes the work of Johannes Schenck and Johannes Jakob Wepfer, two medical scientists in the six-teenth and seventeenth century, whose studies are almost entirely unknown. Johannes Schenck (1530-1598 completed a collection of clinical observations including both reports from antiquity and from his contemporaries. In his first volume (Observationes medicae de capite humano, Basel, 1584 he considers the major diseases of the head and face. At least 16 of these observations refer to patients with aphasia. Johannes Jakob Wepfer (1620-1695 published several texts on neuroanatomy, brain vascularization and apoplexy. He gathered a collection of neurological cases (Observationes medico-practicae de affectibus capitis internis & externis, which was published posthumously. At least 15 of the 222 observations included report the presence of aphasia. As expected, in almost all cases, aphasia was due to lesions in the left hemisphere. Wepfer, however, does not seem to have noted this asymmetry. In the second part of this article, the extraordinary study of Jean-Baptiste Bouillaud (1825 will be reported: Bouillaud outlined the basic principles of anatomo-functional correlations in neuropsychology and applied these principles in order to confirm empirically the role of the frontal lobes in language and speech production.

  4. Valutare gli impatti della qualità sociale e ambientale sul benessere soggettivo

    Marina Bravi

    2016-08-01

    Full Text Available Monetary metric is useful whenever it is necessary a cost/benefit assessment that implies a change of the degree of individuals and families well-being. The work is devoted to investigate and assess the impact of Environmental and Social Quality (ESQu on Subjective Well-Being (SWB at regional scale, keeping under control the individual heterogeneity. After a brief introduction, the application of the Life Satisfaction Approach (LSA on a large data-base of Italian official statistical source, allows establishing the operative practicality of this method through monetization of the impact of some externalities. In this direction and by way of an example, some regional differentials, in terms of environmental and social benefits-costs, are highlighted and their impact on GDP is estimated.

  5. Laurearsi in Toscana - tutti gli articoli Unipi: il Rettore incontra i ricercatori toscani a Ginevra

    2007-01-01

    The students of our region are beyond hundreds who work near CERN, the most important scientific research structure in the world. Among them, professors of university, researchers, engineers and technicians coming from the 3 Tuscan universities. (1/2 page)

  6. La concezione di "mare" presso gli Ittiti tra simbolo e realtà

    Vigo, Matteo

    2012-01-01

    to these people. The Hittite word for sea is well attested in various religious, mythological, historical and political texts. Research to date suggests, however, that the Hittites did not have a word for “island”. This paper will briefly review the perception of water space (symbolic or not) in the Hittite world......, on the basis of cultural interferences and through the analysis of some texts....

  7. Gli effetti dell'attrito sulla stima della disuguaglianza in Italia

    F. Biagi; A. Giraldo; E. Rettore

    2009-01-01

    Il volume contiene i rapporti finali di un'articolata ricerca su alcune significative dimensioni della disuguaglianza nell'Italia di oggi: la povertà e l'evoluzione recente della distribuzione del reddito; la condizione di salute e la sua relazione con le caratteristiche socio-economiche degli individui; le disparità nella disponibilità della casa e nelle situazioni abitative. La ricerca, condotta nell'ambito dell'"Osservatorio sulle disuguaglianze sociali", è stata coordinata da un'equipe in...

  8. Basolateral glycylsarcosine (Gly-Sar) transport in Caco-2 cell monolayers is pH dependent

    Berthelsen, Ragna; Nielsen, Carsten Uhd; Brodin, Birger

    2013-01-01

    Transepithelial di/tripeptide transport in enterocytes occurs via the apical proton-coupled peptide transporter, hPEPT1 (SLC15A1) and a basolateral peptide transporter, which has only been characterized functionally. In this study we examined the pH dependency, substrate uptake kinetics and subst...

  9. Il comune di Cesena e gli Open Geo Data sugli incidenti stradali

    Morena Moretti

    2015-02-01

    Full Text Available Il Comune di Cesena svolge dal 2005 una georeferenziazione degli incidenti stradali sui propri territori, raccog liendo una mole di dati che saranno pubblicati su un portale accessibile liberamente. Scopo del portale è informare i cittadini rispetto ai luog hi ad elevato tasso di incidentalità, alla trasparenza dell’ente rispetto agli interventi di sicurezza stradale e alle azioni di controllo nei period i critici.The Municipality of Cesena held since 2005, a georeferencing of road accidents of its territory. Since January 2015 these data will be published on the new open data portal of the City realized with technology StatPortal Open Data. By acting as "data visualization" these open geo data will be accessible via the web with maps, charts and graphs with multiple benefits, from increased knowledge of citizens compared to places with a high accident rate to the transparency of the institution from the measures road safety and control actions during critical periods. 

  10. Gli usi sociali e politici dell’arte contemporanea fra pratiche di partecipazione e di resistenza

    Ivan Bargna

    2013-10-01

    Full Text Available Se la produzione, circolazione e fruizione di immagini e oggetti esteticamente elaborati ha sempre giocato, sia pur in maniera e misura diversa a seconda dei tempi e dei luoghi, un ruolo importante nella creazione e disarticolazione dei legami sociali, oggi tutto ciò avviene secondo modalità peculiari e inedite che riconfigurano quello che è il campo di studi di un’antropologia estetica e dell’arte.

  11. LE PAROLE DELL’IDENTITA’: GLI ITALIANI VISTI DAI “NUOVI MILANESI”

    Andrea Groppaldi

    2014-02-01

    Full Text Available La lingua dei “nuovi italiani” (in questo caso “nuovi milanesi” è lingua nuova, che si avvale di prestiti dalle lingue d’origine e li innesta in quella italiana. Ma è una lingua curiosa che si avvale anche di espressioni gergali, dialettismi, forme insolite e colorate che appartengono alla lingua non-ufficiale italiana. Esamina così un consistente numero di forme linguistiche tra italiano, dialetto, gerghi, prestiti e frasi idiomatiche, tratti in gran parte dagli autori che hanno partecipato al convegno. Nell’ultima parte del saggio prende in esame l’uso dei nomi e la loro funzione (talora di gabbia nel processo di integrazione delle varie etnìe.  Words of identity: italians viewed by the “new milanesi” The language of “new Italians” (in this case “new Milanesi” is a new language, which uses loans from the languages of origin and adds them to the Italian language. It is a curious language which also uses jargon, dialects, colorful and unusual forms that are part of the non-official Italian language. The paper examines a large number of linguistic forms between Italian, dialect, slang, idioms and loans, collected by the authors who participated in the conference. The last part examines the use of nouns and their function (sometimes like a cage in the integration process of the various ethnic groups.

  12. Gli archivi e l’archivistica tra consapevolezze, illusioni, delusioni e speranze

    Antonio Romiti

    2013-03-01

    Full Text Available The practice of archival recording has undergone massive changes over the last fifteen years, mainly due to the employment of new technologies for gathering and organizing archived items. Economic and social conditions have also changed, to such an extent that state and other kinds of archives have had to struggle in order to adapt to these changes. However, this phenomenon has been part of a much wider process of diversification as regards the functioning and scope of archival structures in Italy. In 1974, the Ministry of Cultural and Environmental Heritage was introduced, an event which was a watershed for broadening archival procedures, and which made it possible to encompass items from private-funded locales/institutions, including a dedicated section on issues of new technologies, which would change the overall context of archival recording. The Europeanization and globalization of forms of dissemination of knowledge have overturned the former setup of State Archives throughout Italy, and this article will address the phenomenon and its effects on the overall state of the art in the practice of archival recording. By taking into account different case studies in the North and South of the country, the article makes proposals for a renewed role of the archivist in State archives ahead of modern technologies.

  13. Crescere nella migrazione. Generi e sessualità fra gli adolescenti di origine straniera

    Federica Tarabusi

    2015-08-01

    Full Text Available How do the children of immigrants define and represent themselves in the Italian society? How do they build and negotiate their own subjectivity of women/men and experience their sexual bodies in adolescence? In what way their everyday experiences are different from (or similar to those of Italian teenagers?Grounded on an ethnographic work carried out in the context of Bologna and surrounding area, the paper investigates the symbolic construction of gender and sexuality among the teenagers of foreign origin. Exploring their multiple biographical trajectories, the research reveals a pattern of resources, subjective desires, imaginaries largely shared with the younger generations, but also their multiple attempts to negotiate and challenge both parent’s expectations and Italian society’s request of “assimilation”. Giving voices to the children of immigrants, the empirical data helps us to deconstruct, through the lens of sexuality, gender and bodily practices, essentialist ideas and stereotypical representations attached to the so-called second generations as well as to discuss the role play by structural inequalities and exclusionary processes in shaping their experience of being adolescents. 

  14. SCENARI E POLITICHE DI DISTRETTO PER LA CITTÀ METROPOLITANA DI REGGIO CALABRIA: GLI ECODISTRETTI

    Maurizio Malaspina

    2014-12-01

    Full Text Available The contribution highlights how the birth of a district, especially with the proliferation of genres that have characterized its recent spread in terms of models, governance, productive sectors, affects fundamentally the creation of the network, an organization aimed at horizontal bridge the gap dimensional optimizing the quality parameters. The creation of sectorial microdistrict in local contexts with strong identity characterization is the proposed road to the construction process of the development scenarios of the Metropolitan City of Reggio Calabria.

  15. Tecnica e riproducibilità: gli esiti della metafisica occidentale e la nuova dimensione estetica

    Antonio Martino

    2015-03-01

    Full Text Available AbstractNihilism coincides with the accomplishment of metaphysics which, in turn, culminates in the triumph of the scientific rationality. The essence of metaphysics, in its last accomplishment (nihilism, consists of the oblivion of being, while the essence of nihilism represents the awareness of that oblivious accomplishment. The shadow of the Being (Seiende covers the Being (Sein while the real facts lead the scientific-experimental thought determining, from Galileo on, the supremacy of the experimentally reproducible objectivity as well as of the scientific method on the actually productive subjectivity. The result is the so-called «technological planetary orientation»: lost the sense of the metaphysic question, philosophy comes to the end. In other words, after Nietzsche there is only science/technique. Therefore, how has all that weighed on the aesthetic dimension? The artistic productivity, though reflecting the torment of the human soul, towards the nothing of sensemiraculously tends to make the light of the Being (Sein appear from the shadow of the material Being (Seiende. Every originating artistic shine of the Being (Seinis an aesthetic expression from the pure creative human act which celebrates its own being as a dynamic bond of connections (inter-essewould say evocatively Kierkegaard-Climacus that cannot understand the origin but spread its form. Der Nihilismus entspricht der Erfüllung der Metaphysik, die wiederum im Triumph der wissenschaftlichen Rationalität. Das Wesen der Metaphysik, in seiner extremen Erfüllung (Nihilismus, besteht aus dem Vergessen des Seins und das Wesen des Nihilismus ist das Bewusstsein solcher vergessender Erfüllung. Der Schatten des Seiendes verdeckt das Sein und die Elemente der realen Welt (die Tatsachenlenken das wissenschaftliche, experimentelle Denken, das seit Galileo, die Hegemonieder experimentell reproduzierbaren Objektivität, und der wissenschaftlichen Methode über die effektive Subjektivität auslöst. Das Ergebnis ist die sogenannte „technologische/planetarische Ordnung“: geht der Sinn der metaphysischen Frage verloren, kommt die Philosophie zu einem Ende. Mit anderen Worten, nach Nietzsche gibt es nur Wissenschaft/Technik. Nun, wie hat all dies die ästhetische Dimension beeinflusst? Obwohl die künstlerische Produktivität das Leiden der menschlichen Seele angesichts des Nichts des Sinns wiederspiegelt, so neigt sie doch dazu, das Licht des Seins (wie durch ein Wunder aus dem Schatten des materiellen Seiende sichtbar werden zu lassen. Die Quelleeines jeden künstlerischen Schaffens des Wesens ist die ästhetische Manifestation des reinen kreativen Akt des Menschen, der  das eigeneSein als dynamischen Kern von Beziehungen zelebriert (inter-esse würde Kierkegaard-Climacus sagen der nicht im Stande ist, den Ursprung zu erfassen, sondern nur dessen Formen verbreiten.

  16. Com'è profondo il mare Gli uominiche lo misurano, l'Istituto Idrografico della Marina

    Alessandro Nobili

    2014-03-01

    Full Text Available Measuring sea depths: the Italian hydrographic InstituteSince 1873 the Italian Hydrographic Institute (Istituto Idrografico della Marina - IIM has been involved in sea depth measuring in order to improve safety of navigation. The main mission of the institute is to provide hydrographic services for waters of Italian national responsibility required by the International Convention for the Safety of Life at Sea (SOLAS. This role requires the coordination and determination of policy and standards which covers both hydrographic surveying and charting, as well as contributing tothe coordination, exchange and standards related to geospatial data in general. IIM’s traditions and methods take their origins from the pioneers of hydrography; in this paper the author briefl y describes the main stages in IIM’s history, passing throught scientifi c and international activities and concludine with challenges and prospects of digital era.

  17. Il giudice Wyndham e gli spettacoli per l’intrattenimento di Elisabetta I a Norwich

    F. Minetti

    2012-08-01

    Full Text Available ENThe fabrication of a popery charge, which in August 1578 Edward Rokewood was subjected to while hosting Queen Elizabeth in the East Anglia, has been dubiously dealt with in the field of gender studies (Dovey, 1996; Brownlow, 2003 or overrated by historians of Puritanism to corroborate how the queen’s Privy Council ruled the kingdom through a manipulation of her weak will and female simple-mindedness (MacCulloch, 1986; Collinson, 2007. My paper proposes to substantiate how the intrigue against Rokewood arose from a wider social group of officers than the Lord Chamberlain Sussex and the royal favourite Leicester’s clique. Some days after the papist had been jailed in the castle of Norwich, mayor and aldermen commissioned public speeches and a composite biblical pageantry which, though allusively, both staged Rokewood’s conspiracy in favour of the Spanish king’s invasion and urged Elizabeth to inflict a death penalty. But the queen did not only turn a deaf ear to these pressures, presiding over the trial proceedings to get a time of forced reform for Rokewood. Some letters from the judge of assizes Francis Wyndham will help me to suggest that she was nor manipulated neither prevented from seeing that the case had been invented by the municipal corporation to advoke their bishop’s means of control and persecution of dissent.ITL’invenzione di un’accusa di papismo, che ad agosto del 1578 Edward Rokewood subisce mentre ospita la regina Elisabetta nella regione orientale dell’Anglia, è discussa con grande scetticismo nel settore dei gender studies (Dovey, 1996, pp. 54-55; Brownlow, 2003, pp. 11-13, oppure sopravvalutata da storici della cultura puritana convinti che il Privy Council tenga le redini del regno manipolando volontà e credulità della regina (MacCulloch, 1986, pp. 218-219; Collinson, 2007, p. 131. Questo paper si propone di documentare come la macchinazione ai danni di Rokewood si estenda ben oltre la cerchia del ciambellano di corte Sussex o del favorito regio Leicester. Infatti, alcuni giorni dopo che il papista veniva imprigionato a Norwich, la fanatica municipalità commissiona orazioni pubbliche ed uno spettacolo di sacre rappresentazioni che, sia pure in maniera allusiva, drammatizzano come Rokewood cospiri a favore dello sbarco di Filippo II sull’isola e istigano Elisabetta a comminare una condanna a morte. La destinataria non solo resiste alle pressioni giustizialiste, ottenendo che il processo si concluda con un periodo di arresto ‘riabilitativo’. Alcune lettere del giudice di assise mi aiuteranno a segnalare come, a dispetto della corrente storiografia, Elisabetta non è né manipolata né ignora che il caso di eresia sia stato inventato per avocare il potere di sorveglianza e punizione del dissenso che il vescovo è titolato ad esercitare nella diocesi.

  18. Gaimar, Wace e gli altri autori. La traduzione alle origini della letteratura anglo-normanna

    Margherita Lecco

    2015-05-01

    Full Text Available L’articolo traccia un breve profilo del passaggio dall’impiego della lingua latina ed anglo-sassone presso le corti dei re Normanni d’Inghilterra alla lingua anglo-normanna per la stesura di opere dapprima storiche, poi propriamente letterarie. L’uso del volgare, che si avvia con Geffrei Gaimar intorno al 1140, viene mediato dalla traduzione dei testi che egli fa traendo la propria materia dalle opere degli storiografi attivi nel decennio precedente (Guillaume de Malmesbury e Geoffrey de Monmouth ma anche l’Anglo-Saxon Chronicle. Da latino a volgare, e da volgare (anglo-sassone e gallese a volgare, la letteratura anglo-normanna, attraverso l’incrocio e la traduzione da più lingue, viene definendo un corpus di tradizioni e di testi davvero cospicuo nell’età medievale europea. Dalla tradizione anglo-normanna, tuttavia, una letteratura ulteriore viene ad avviarsi, quella middle-english, che riprende l’english poco praticato in età normanna, pronto a risorgere come lingua non solo popolare ma anche letteraria, benché attraverso la ripresa, con un ulteriore processo di traduzione, di temi e testi anglo-normanni. This article briefly outlines the passage from the use of Latin and Anglo-Saxon to the Anglo-Norman language for writing first historical, then also literary works, at the court of the Norman kings of England. The vernacular was first used for literary creation by Geffrei Gaimar (around 1140, who took inspiration from and translated part of the Anglo-Saxon Chronicle and other works of historiographers active in the previous decade (Guillaume de Malmesbury and Geoffrey de Monmouth. The corpus of texts and traditions of Anglo-Norman literature – which is rather conspicuous when compared to others of medieval European literature – generated from intersections and translations of several languages: mainly Latin, Anglo-Saxon and Welsh. The Anglo-Norman literary tradition would, in its turn, pave the way for a new literature in Middle English, which, although scarcely used during the Norman age, would later become not only the language of common people, but also that of literary production.

  19. Aristotele, Einstein e gli altri i grandi scienziati tra pensiero e vita quotidiana

    Fischer, Ernst Peter

    1997-01-01

    Abbiamo spesso un'idea confusa di quelle che sono state le vite degli uomini che hanno consacrato all'impresa scientifica le loro migliori energie. Eppure, non solo nell'arte e nella filosofia, ma anche nella scienza "lo stile è l'uomo" e la psicologia del singolo scienziato può rivelarsi il miglior aiuto per capire la logica della scoperta scientifica. Questa è una "storia della scienza raccontata agli amici davanti al caminetto", come l'autore stesso definisce la propria opera. Che non è soltanto una ricostruzione puntuale delle svolte principali che hanno portato alla scienza contemporanea, ma un racconto delle vicende personali di tutti quegli scienziati che sono stati divorati dalla passione del conoscere.

  20. Il comune di Cesena e gli Open Geo Data sugli incidenti stradali

    Morena Moretti

    2015-02-01

    Full Text Available Il Comune di Cesena svolge dal 2005 una georeferenziazione degli incidenti stradali sui propri territori, raccog liendo una mole di dati che saranno pubblicati su un portale accessibile liberamente. Scopo del portale è informare i cittadini rispetto ai luog hi ad elevato tasso di incidentalità, alla trasparenza dell’ente rispetto agli interventi di sicurezza stradale e alle azioni di controllo nei period i critici. The Municipality of Cesena held since 2005, a georeferencing of road accidents of its territory. Since January 2015 these data will be published on the new open data portal of the City realized with technology StatPortal Open Data. By acting as "data visualization" these open geo data will be accessible via the web with maps, charts and graphs with multiple benefits, from increased knowledge of citizens compared to places with a high accident rate to the transparency of the institution from the measures road safety and control actions during critical periods.

  1. A new case of dysfibrinogenemia A.alpha. 13 Gly/Glu

    Kotlín, R.; Reicheltová, Z.; Mal, M.; Suttnar, J.; Riedel, Tomáš; Salaj, P.; Pohlreichová, V.; Geierová, V.; Dyr, J. E.

    2009-01-01

    Roč. 94, č. 2 (2009), s. 676 ISSN 0390-6078. [Congress of the European Hematology Association. 04.06.2009-07.06.2009, Berlin] R&D Projects: GA AV ČR KAN200670701 Grant - others:Internal Grant Agency of the Ministry of Health(CZ) NS9636-3/08 Institutional research plan: CEZ:AV0Z40500505 Keywords : fibrinogen * blood coagulation Subject RIV: CD - Macromolecular Chemistry

  2. FORS, Fiber Optics Reflectance Spectroscopy con gli spettrometri miniaturizzati per l’identificazione dei pigmenti

    Antonino Cosentino

    2014-01-01

    Full Text Available AbstractQuesto articolo riporta i risultati del test di un sistema FORS (Fiber Optics Reflectance Spectroscopy assemblato con componenti Ocean Optics. Questa ditta ha sviluppato una linea di spettrometri miniaturizzati e dal costo estremamente contenuto. Le trascurabili dimensioni e peso di questo sistema FORS lo rendono uno strumento assolutamente adatto alle indagini dell’arte in cantiere. Sono stati testati 54 pigmenti storici in polvere e stesi con gomma arabica, tempera all’uovo, olio e affresco. Il sistema è stato provato su un quadro dell’800 e su opere murali del barocco siciliano.   This paper shows the results of the testing of a FORS (Fiber Optics Reflectance Spectroscopy system assembled with Ocean Optics components and featuring a USB4000 miniaturized and low cost spectrometer. The small dimensions and little weight of this FORS system make it adapt for the specific needs of art examination in the field. 54 historical pigments have been tested both in powder and laid with gum Arabic, egg tempera, linseed oil and fresco. The FORS system was also tested on a 1800s oil painting and on 1700s Sicilian baroque murals and the results compared with multispectral imaging analysis.   

  3. PGC-1alpha Gly482Ser polymorphism associates with hypertension among Danish whites

    Andersen, Gitte; Wegner, Lise; Jensen, Dorit Packert

    2005-01-01

    in combination with previously published results. The Ser/Ser genotype was significantly associated with a reduced risk of hypertension and with lower systolic, diastolic, and mean arterial blood pressure levels, predominantly among women. Finally, in a combined analysis using data obtained in both sexes...... to elucidate the differential role of this polymorphism in men and women.......PGC-1alpha is a coactivator of numerous transcription factors and is expressed in tissues with high energy demands and abundant in mitochondria. It is induced in the myocardium on fasting and physical exercise, and cardiac-specific overexpression stimulates mitochondrial biogenesis in mice...

  4. Crystal structure of raw pure Mysore silk fibre based on (Ala-Gly)2 ...

    Unknown

    2005-03-23

    Mar 23, 2005 ... at sites 1′ and 2′ by the 2-fold screw symmetry, respec- tively. ..... Demura M, Minami M, Asakura T and Cross T A 1998 Structure of Bombyx more ... A34 3–11. Spek A L 2003 Single-Crystal structure validation with the pro- ... Takahashi Y, Gehoh M and Yuzuriha K 1999 Structure refinement and diffuse ...

  5. Literature in Focus - Gli anelli del sapere = The Rings of Knowledge + The LHC bible

    Tullio Basaglia

    2010-01-01

    A fascinating photographic survey describes the essential phases of constructing the colossal equipment to build the four main experiments. It is enriched by a sophisticated, experimental graphic design and the stories of the Italian scientists from the INFN (National Institute of Nuclear Physics), who made vital contributions to the project and constructing the LHC. The book is completed by descriptions illustrating the important contribution that Italian companies made to this major project, demonstrating Italy’s competitive capacity for excellence in the most important technological undertakings in Europe. Edited by Federico Brunetti (Politecnico di Milano) Published by Editrice Abitare Segesta The presentation of the book will take place on Tuesday, 9th March, 4.00 p.m. in the Pas Perdus area. At the same time, the INFN photo exhibition installed in the occasion of the International Women's day will also be inaugurated. A reception will be sponsored by the Permanent Mission of Italy to th...

  6. U.S. EPA, Pesticide Product Label, GLY N GO, 05/11/2009

    2011-04-21

    ... conjunction with tillage practices in fallow systems or preplant ... natural rainfall or via a sprinkler system. ... I~~~~ T~kSE~~~i~~~ ~~VDE~~~~~Rt~~~~E ...

  7. Inserti abusivi e attribuzioni indifendibili. Spigolando tra gli unica del canzoniere provenzale Sg

    Giuseppe Tavani

    2015-05-01

    Full Text Available The central section of the Occitan songbook kept at the Biblioteca de Catalunya (ms. 146, generally known as Sg since it originally came from Saragossa (from Gil y Gil’s library, continues to prove to be of great interest for the study of the reception of troubadour poetry in Catalan milieux, at a time when a new poetic tradition was taking shape, influenced by the Toulouse tradition of the Consistori de la Gaia Sciensa. These particular circles and period naturally influenced the selection of authors and texts – with the resulting inclusions and exclusions – as well as the accompanying attributions. The latter may in part be due to a tradition that was already corrupt, but also, especially as far as the unica are concerned, to a form of mimetic parasitism, accompanied by a tendency for illicit inclusions in the case of some particular texts for which, as in previous articles, I have tried to point out the degree of ‘impertinence’ and to decide whether to add them to the list of suspects, to place them under interrogation, or – in the most clearly arbitrary cases – to simply remove them. This paper will focus on a planh attributed to Raimbaut de Vaqueiras (394,4a, which Sg shares with Ve-Ag, and on three unica which Sg alone attributes to Giraut de Borneil (242,18a; 52a; 69a.

  8. A novel fibrinogen variant - Praha I: hypofibrinogenemia associated with .gamma. Gly351Ser substitution

    Kotlín, R.; Chytilová, M.; Suttnar, J.; Salaj, P.; Riedel, Tomáš; Šantrůček, J.; Klener, P.; Dyr, J. E.

    2007-01-01

    Roč. 78, č. 5 (2007), s. 410-416 ISSN 0902-4441 Institutional research plan: CEZ:AV0Z40500505 Keywords : fibrinogen * missense mutation * hypofibrinogenemia Subject RIV: CD - Macromolecular Chemistry Impact factor: 2.163, year: 2007

  9. Activity-Based Intelligence prevedere il futuro osservando il presente con gli strumenti Hexagon Geospatial

    Massimo Zotti

    2015-06-01

    Full Text Available The intelligence of human activities on the earth's surface, obtained through the analysis of earth observation data and other geospatial information, is vital for the planning and execution of any military action, for peacekeeping or for humanitarian emergencies. The success of these actions largely depends on the ability to analyze timely data from multiple sources. However, the proliferation of new sources of intelligence in a Geospatial big data scenario increasingly complicate the analysis of such activities by human analysts. Modern technologies solve these problems by enabling the Activity Based Intelligence, a methodology that improves the efficiency and timeliness of intelligence through the analysis of historical, current and future activity, to identify patterns, trends and relationships hidden in large data collections from different sources.

  10. L’esercizio e gli Essais. La retorica filosofica di Montaigne

    Emanuele Ronchetti

    2012-06-01

    Full Text Available The Essais’s writing generates a form of original pyrrhonian philosophical discourse, apt to express the infinite variety of the world. In this way it paves the way to a limitless method of investigation: zetetics.

  11. Supply Chain Optimization of Integrated Glycerol Biorefinery: GlyThink Model Development and Application

    Loureiro da Costa Lira Gargalo, Carina; Carvalho, Ana; Gernaey, Krist

    2017-01-01

    To further advance the development and implementation of glycerol-based biorefinery concepts, it is critical to analyze the glycerol conversion into high value-added products in a holistic manner, considering both production as well as the logistics aspects related to the supply chain structure...... is able to identify operational decisions, including locations, capacity levels, technologies, and product portfolio, as well as strategic decisions such as inventory levels, production amounts, and transportation to the final markets. Several technologies are considered for the glycerol valorization...... to high value-added products. Existing countries with major production and consumption of biodiesel in Europe are considered as candidates for the facility sites and demand markets, and their spatial distribution is also carefully studied. The results showed that (i) the optimal solution that provides...

  12. ll social ed il mobile commerce secondo gli esperti di digital marketing

    Latorre Annaluce; Vernuccio Maria

    2014-01-01

    Advances in social media and mobile communication have ledto the development of "social" and "mobile commerce". Stemming from aliterature review, this paper aims to explore practitioners' interpretative perspective of the phenomena. To this purpose, we have drawn upon 25 in-depth interviews with Italian digital marketing specialists and examined their point of view using content analysis techniques. The findings show the characteristics, potentialities and critical aspects of "social" and "mo...

  13. Legami tra pianure. Gli intermediari nella migrazione dei panjabi indiani in Italia

    Azzeruoli, Vanessa

    2014-01-01

    Connected plains The brokers within the panjabi indian migration in Italy This research contributes to the field of migration studies by investigating the social construction of the brokers in Indian Punjabi migration in Italy. The thesis analyzes the influence of networks during the migration process and its relationship with the migration policies framework and economic conditions. The purpose of the investigation is to examine in depth the multidimensionality of transna...

  14. Gli strumenti di contrasto al terrorismo e al cyber-terrorismo nel contesto europeo

    Carolina Lamberti

    2014-07-01

    Full Text Available This article is a review of European policies towards the fight against terrorism and cyber-terrorism that has been operating since 2001, the year of the win Tower's attack in New York. Its aim is to identify changes occurred after the major terrorist attacks of the beginning of this century (Madrid in 2004, London in 2005, Oslo and Utøya in 2011, Toulouse and Montauban in 2012, Boston in 2013. Starting from the international academic debate about the definition of terrorism and cyber-terrorism, the article highlights the current traits of this phenomenon, with an interest towards the massive effects of its technological evolution. The new distinguishing marks of terrorism, different from the past, have imposed the development of new strategies at national and international levels in order to fight against it. Afterwards, the article focuses on Italian and European policies, underlying the problems regarding the delicate balance between security needs and protection of basic freedoms and democracy.

  15. Gli incerti confini del cosmo dai buchi neri alle macchine del tempo

    De Felice, Fernando

    2000-01-01

    Il libro mostra come la gravitazione abbia un ruolo determinante nei fenomeni cosmici che più ci impressionano, da quelli relativamente vicini alla nostra esperienza (come la formazione delle stelle) a quelli meno intuibili (come la formazione dei buchi neri), alla struttura dell'Universo nel suo insieme. Oltre a spiegare, in modo qualitativo, concetti come la geometria dello spazio-tempo, la relatività del tempo e i coni di luce, l'autore formula un'ipotesi innovativa tendente a spiegare l'origine della gravitazione e con essa quella dei fenomeni circondati ancora da un alone di mistero: la materia oscura dell'Universo e il destino della materia alla fine del collasso gravitazionale.

  16. Real Estate Crowdfunding. Un’innovazione finanziaria per gli investimenti immobiliari

    Giacomo Morri

    2016-12-01

    Full Text Available Questo articolo presenta una panoramica del crowdfunding e delle sue potenziali applicazioni al settore immobiliare. La crisi economica globale del 2008 ha condotto a una paralisi dell’economia che ha comportato la riduzione della fiducia nel sistema bancario e nei servizi finanziari in generale. Nell’ultimo decennio una delle tecniche di finanziamento più rivoluzionaria è stata il crowdfunding, cioè la possibilità di essere finanziati dalla “folla” (crowd attraverso l’utilizzo di Internet. Il crowdfunding nel settore immobiliare ha origine negli Stati Uniti nel 2012 con il Jumpstart Our Business Start-up Act (JOBS Act, dove da allora si sta sviluppando a ritmi elevati. In Italia, il crowdfunding per progetti nel settore immobiliare non si è ancora sviluppato in quanto la legge in concreto non ne consente l’utilizzo; tuttavia esistono alcune piattaforme che stanno utilizzando il concetto in modi differenti, raccogliendo un certo interesse. Il Real Estate Crowdfunding può rappresentare un’interessante opportunità per i privati che vogliono investire nel settore immobiliare, ma non hanno abbastanza capitale per partecipare a grandi transazioni o esperienza nella gestione. In quest’articolo, dopo un’iniziale descrizione del crowdfunding in generale, ci si focalizza sulla sua applicazione nel settore immobiliare, analizzando il mercato statunitense dove vi sono le piattaforme più sviluppate. Infine, si presenta una panoramica del mercato italiano, spiegando le normative e i limiti del crowdfunding e descrivendo la modalità operativa sviluppata da alcune piattaforme per superare legalmente tali limitazioni. Negli Stati Uniti sono già operative oltre 125 piattaforme con una raccolta superiore a un miliardo di dollari; in Italia invece ci sono ancora limiti per le start-up che si occupano di crowdfunding e sono state create solo poche piattaforme che operano con un modello parzialmente diverso.

  17. ethyle ling gr ) R. Br. d ne gly rowth o .] and drough col

    SAM

    soil layers. Drought constitutes one of the most important environmental stress factors limiting plant growth. (Sivakumar, 1992). Large areas of drylands are threatened unproductive due to human activities and ... molecular weight is inert, non-ionic. They are small enough to influence the osmotic potential, but large enough ...

  18. Modernità ‘primitivista’: dopo gli indiani, i neri.

    Sandra Jatahy Pesavento

    2009-05-01

    Full Text Available The concept of primitive, besides that one of identity that cross it, is a social construction, a collective representation founded in a relationship between a “we” and a “others”. It's not possible to contend that a people is primitive in his own nature, because this concept is built with images and discourses, narratives, that put in contrast the world of those ones that analyze and those ones that are analyzed. In this way, the paper presents the discussion – in the process of construction of a Brazilian national identity, between the Nineteenth and Twentieth centuries – concerning the sign of the primitive that brand the Indians and Black people presence in this formative path of “Brazilianity”. Beginning with a negative perception about the difference, in the Nineteenth Century thought, its comes – in the Twenties – to a positive understanding about the Brazilian cultural diversity, in the modernism ideas and in the Gilberto Freyre's analysis. The Freyre sociological approach allow us to think about the frontiers present in a world without frontiers and show us that the boundaries – as imaginaries constructions of the society – are part of the social imaginary.

  19. Characterization and microstructure of HPMC/Gly:AgNO3 polymer composites

    Ananda, H. T.; Urs, G. Thejas; Somashekar, R.

    2018-04-01

    This study reports the synthesis and characterization of AgNo3 doped HPMC/Glycerol blend films. The microstructural parameters of these composites were evaluated employing whole powder pattern fitting method (WPPF) and the results obtained are related with other physical properties. AC conductivity results and optical band gap evaluated from UV/Vis studies are focused to establish structure property relations. These composite films are bio-degradable in nature and non-hazardous, this makes them very suitable candidates for applications in appropriate fields.

  20. Aspetti tributari dei crediti d’impresa contabilizzati secondo gli Ias/Ifrs: prime riflessioni sistematiche

    A. CONTRINO

    2011-01-01

    1. La disciplina dei crediti d’impresa nei principi contabili internazionali. – 1.1. Costo ammortizzato, valore di presumibile realizzo e «perdite da valutazione». – 1.2. Derecognition, «realizzo» ai fini IAS/IFRS e «perdite da cancellazione». – 2. I nuovi termini di rilevanza fiscale dei crediti IAS/IFRS in forza del principio di derivazione «rafforzata». – 2.1. Sui confini dell’area di determinazione dell’imponibile riservata dall’art. 83 ai principi contabili internazionali. – 2.2. Il nuov...

  1. Gli strumenti per la valutazione soggettiva del rischio stress lavoro-correlato

    Conway, Paul Maurice

    2009-01-01

    The instruments for subjective assessment of work-related stress are aimed to evaluate individual perceptions of aspects that are crucial in the causal process linking psychosocial exposure to harmful physical and psychological health effects. Subjective assessment is most commonly performed...... of the above-mentioned instruments are depicted. Finally, important aspects relating to the usage of self-administered questionnaires are emphasized, mainly when field evaluations are undertaken in specific work contexts......., those that have been applied to a higher number of subjects in Italy are the Job Content Questionnaire, the Effort/Reward Imbalance Questionnaire, the Occupational Stress Indicator and the Multidimensional Questionnaire of Organizational Health. In this contribution, the main characteristics...

  2. uno sguardo sul colonialismo italiano: gli scritti di erminia dell'oro

    User

    un'esperienza vissuta accompagnando il mio nipotino al museo di. Storia Naturale di Milano. Da allora ho continuato a scrivere per i ragazzi l'ultimo mio libro s'intitola Il gatto rapito, edito dal Battello a. Vapore”. Nonostante abbia lasciato l'Eritrea a vent'anni e da quel momento abbia sempre vissuto a Milano, nel corso della ...

  3. ArcGIS 9.3 ed i database spaziali: gli scenari di utilizzo

    Francesco Bartoli

    2009-03-01

    Full Text Available ArcGis 9.3 and spatial databases: application sceneriesThe latest news from ESRI suggests that it will soon be possible to link to the PostgreSQL database. This resulted in a collaboration between the PostGis geometry model with SDOGEOMETRY - the oracle database - a hierarchial and spatial design database. This had a direct impact on the PMI review and the business models of local governments. ArcSdewould be replaced by Zig-Gis 2.0 providing greater offerings to the GIS community. Harnessing this system will take advantage of human resources to aid in the design of potentconceptual data models. Further funds are still requiredto promote the product under a prominent license.

  4. “Storm Landscape”: from the reality effect to the moralized mimesis. The examples of Apollonius Rhodius and Quintus of Smyrna. "Le paysage de tempête" : de l'effet de réel à la mimesis moralisée. Les exemples d'Apollonios de Rhodes et de Quintus de Smyrne 'Il paesaggio della tempesta': dall'effetto realistico alla mimesi moralizzante. Gli esempi di Apollonio Rodio e Quinto Smirneo

    Laury-Nuria André

    2013-05-01

    non plus par un « paysage en mouvement » présentant de manière pathétique la violence et la mort, mais par un pathos qui n’est plus perceptible qu’à travers l’usage d’un vocabulaire axiologique et philosophique ? Cette communication entend donc montrer comment le concept de mimesis évolue d’une acception d’imitation, d’effet de réel, à une compréhension plus éthique, moralisée à la fin de l’antiquité. Le paysage constitue donc bien un des lieux d’observation privilégiés de la mise en œuvre d’une mimesis évolutive à la période hellénistique et tardive.La rappresentazione del paesaggio antico nell’epica ellenistica e imperiale è un ambito di indagine privilegiato per l’esame delle funzioni e dell’evoluzione dell’antico concetto di imitazione. Un paesaggio epico in particolare, quello del mare in tempesta, occuperà la nostra attenzione: il passaggio delle Simplegadi in Apollonio Rodio (Argonautica II, 549-610 e la tempesta con cui si concludono i Posthomerica di Quinto Smirneo offrono uno dei più rilevanti studi comparativi. Il paesaggio delle Simplegadi, infatti – un paesaggio estremo – è luogo ove tutti gli elementi risultano capovolti e il poeta descrive una scena di caos, prossima all’irrappresentabilità. Eppure, rileggendo il passo, risulta che esso è tutt’altro che caotico: costruito attentamente a partire da un ritmo narrativo regolare e organizzato in tre fasi che imitano i moti delle onde, esso crea un effetto realistico ‘rassicurante’ dato da una prosodia e da uno stile che seguono Omero molto da vicino. Mimesis, effetto realistico e riscrittura omerica sono strettamente associati in questo tempestoso paesaggio apolloniano.Tuttavia, per la tempesta Quinto presenta un paesaggio dalla differente tessitura mimetica: la tempesta che conclude i Posthomerica, che intendono essere una riscrittura dell’Iliade e dell’Odissea, dipende dall’intertesto ellenistico molto più che da quello

  5. Protesta dei ricercatori contro gli editori: politiche di prezzo insostenibili per la ricerca. Nel mirino Elsevier e Springer

    Maria Chiara Pievatolo

    2012-02-01

    Full Text Available Ricopio, per chi non segue il nostro servizio Twitter, la notizia uscita ieri su Ciber Newsletter, su PLEIADI e altrove.  I mostruosi margini di profitto delle multinazionali dell’editoria scientifica sono riportati qui. Aggiornamento 2/2/2012:  la risposta di Springer è visibile qui; quella di Elsevier qui.  Qui c’è un’analisi dei loro argomenti. Aggiornamento 3/2/2012:  l’Economist si [...

  6. Gli interventi nazionali sui problemi della migrazione: strategie e programmi (National interventions on the issues of migration: strategies and programs

    Carmine CLEMENTE

    2016-03-01

    Full Text Available Europe has become the first continent of attraction for migrants and is also the first continent as for incidence of the population emigrated to the total of population. In Italy, in particular, the foreign population has grown from 2000 to 2013 at an average rate of 10% per annum. This scenario, plus the emergence of immigrant landings on the Italian coast as a point of arrival but also as a point of departure to other European countries, leads to awareness of the need for answers, not only at the Italian institutional level but also within a European convergence framework, in the search of policies and programs most appropriate to a new social context, strongly characterized by the presence of the immigrant population

  7. Ebdomadari versus canonici: gli istituti clericali, il potere ecclesiale e la topografia medievale del complesso episcopale di Napoli

    Lucherini, Vinni

    2006-12-01

    Full Text Available

    This article suggests a new interpretation of a series of medieval records (X-XIV centuries, in which the congregatio ecclesiae Sanctae Restitutae and the congregatio Salvatoris, are mentioned, both congregations of priests attached to Naples Cathedral. The authoress reaches the conclusion that these were not two chapters of canons corresponding to two different cathedral buildings: Santa Restituta and the Salvatore, but rather two hierarchically different congregations within the same Cathedral. The members on one of these congregations -from 1337 officially called Ebdomadari-, in the 18th century, invented the tradition of a second old Cathedral, for economic reasons and to increase their power.



    In questo articolo si propone una nuova interpretazione di una serie di documenti medievali (X-XIV secolo menzionanti le congregazioni clericali della Chiesa cattedrale napoletana, la congregatio ecclesiae Sanctae Restitutae e la congregatio Salvatoris. Le conclusioni a cui si giunge è che non si tratta di due Capitoli di Canonici facenti capo a due distinti edifici cattedrali, Santa Restituta e il Salvatore, ma di due congregazioni gerarchicamente differenziate all’interno della medesima Cattedrale, i membri di una delle quali, ufficialmente chiamati dal 1337 Ebdomadari, inventarono nel Settecento, per ragioni di denaro e di potere, la tradizione di un seconda antica Cattedrale.

  8. Gli spazi marginali di un personaggio bizzarro: l’adattamento del romanzo Totò il buono

    Matteo Martelli

    2012-11-01

    Full Text Available Il saggio propone uno studio del romanzo di Cesare Zavattini, Totò il buono, e della riscrittura filmica Miracolo a Milano (firmata da Zavattini stesso e Vittorio De Sica attraverso l’analisi della figura del protagonista, Totò, personaggio eccentrico e buono. In maniera particolare, a partire dalla presenza in entrambi i testi di spazi di marginalità culturale dai quali si costruiscono le qualità bizzarre del personaggio, il saggio descrive come il lavoro d’adattamento rielabori le formulazioni del romanzo attraverso l’introduzione di un punto di vista della tradizione, estetica e antropologica, del grottesco e del realismo popolare, integrando e spostando al tempo stesso il precedente stato di marginalità da uno spazio asociale alla valorizzazione, sul piano immaginario, di un’utopia comunitaria.

  9. Gli uccelli del Parco Nazionale della Val Grande e delle aree limitrofe (Verbano Cusio Ossola, Piemonte, Italia

    Fabio Casale

    2014-12-01

    Full Text Available Il Parco Nazionale della Val Grande è un’area montuosa protetta sita nelle Alpi, nel Piemonte nord-orientale, Italia. L’area è stata designata quale Parco Nazionale nonché ZPS – Zona di Protezione Speciale secondo la Direttiva “Uccelli” e IBA – Important Bird Area secondo BirdLife International. L’area di studio ha una superficie di 45.873 ha che comprende il Parco Nazionale e le limitrofe Valle Intrasca, Valle Cannobina e Val Loana, i cui ambienti principali sono costituiti da vaste foreste di latifoglie e di conifere, ambienti rocciosi, brughiere, arbusteti e praterie alpine. Le cime più alte raggiungono i 2.300 m s.l.m. La maggior parte della sua superficie è selvaggia e disabitata. In questo lavoro viene riportato un elenco commentato degli uccelli noti per l’area e vengono inoltre forniti dati di sintesi relativi alle attività di monitoraggio condotte in anni recenti (2009-2014. I dati sono stati raccolti sia attraverso revisione bibliografica sia in forma originale, attraverso attività di ricerca sul campo. Queste ultime sono state condotte utilizzando i seguenti metodi: mappaggio dei territori (uccelli nidificanti degli ambienti aperti, punti d’ascolto (uccelli nidificanti, transetti lineari con uso di playback (rapaci notturni, Caprimulgus europaeus, Dryocopus martius, ricerca di campo non standardizzata in ambienti idonei alla nidificazione (Passeriformi nidificanti degli ambienti rocciosi, censimento primaverile di rapaci migratori lungo la rotta migratoria del Lago Maggiore e censimento autunnale di Passeriformi migratori attraverso i passi alpini. L’area ospita o ha ospitato 138 specie di Uccelli delle quali 97 nidificanti, 28 di interesse comunitario (Allegato I della Direttiva “Uccelli”, 42 SPEC - Species of European Conservation Concern secondo BirdLife International. L’area è di grande importanza per numerose specie di uccelli, specialmente per la nidificazione di specie di interesse conservazionistico legate ad habitat montani, quali Bonasa bonasia (20-50 coppie, Tetrao tetrix (40-100 maschi adulti, Alectoris graeca saxatilis (20-50 coppie, Circaetus gallicus (4-5 coppie, Aquila chrysaetos (6-7 coppie, Falco peregrinus (4-5 coppie, Bubo bubo (4 coppie, Aegolius funereus, Dryocopus martius (30 coppie, Monticola saxatilis (5-10 coppie, Tichodroma muraria (5-10 coppie, Pyrrhocorax graculus (50-70 coppie, Prunella collaris (10-20 coppie, Emberiza cia (200-300 coppie, per la migrazione primaverile dei rapaci diurni (15 specie rilevate, delle quali 10 di interesse comunitario, soprattutto Pernis apivorus (21,3% degli individui censiti, Falco tinnunculus (16,7% e Buteo buteo (14,5% e per la migrazione autunnale dei Passeriformi (30 specie rilevate, soprattutto fringillidi (46,9% degli individui censiti, irundinidi (32,8% e motacillidi (15,6%. Gypaetus barbatus e Gyps fulvus vi vengono occasionalmente rilevati.

  10. «Vivere da soldato senza essere tale». Gli scritti di guerra di Thomas Mann

    Francesca Tucci

    2013-11-01

    Full Text Available This essay examines the semantic area linked to military activity and in particular the conditions of the soldier in the essays written by Thomas Mann during the First World War. These reconstructions of a historical theme intermix with contributions about current public affairs, and have as their object the events linked to Germany’s entering the war and the general favour with which this was received. Moreover, warlike metaphors tend to slip comfortably into the profound core of Mann’s writing, since they lend themselves rather effectively to descriptions of the artist’s actual aptitudes and duties.

  11. CtBP1/BARS Gly172 → Glu mutant structure: Impairing NAD(H)-binding and dimerization

    Nardini, Marco; Valente, Carmen; Ricagno, Stefano; Luini, Alberto; Corda, Daniela; Bolognesi, Martino

    2009-01-01

    C-terminal binding proteins (CtBPs) are multi-functional proteins involved in nuclear transcriptional co-repression, Golgi membrane fission, and synaptic ribbon formation. Binding of NAD(H) to CtBPs promotes dimerization. CtBP dimers act as a scaffold for multimeric protein complex formation, thus bridging transcriptional repressors and their targets in the nucleus. Based on size-exclusion chromatography experiments and on the crystal structure of the NAD(H)-free G172E CtBP mutant, we show here that absence of NAD(H) induces flexibility/backbone conformational changes at the dimerization interface and at the CtBP interdomain region. The results presented shed first light on the correlation between NAD(H)-binding and functional CtBP dimerization.

  12. La mente mente, il volto no. Gli indizi dell’inganno nei rapporti interpersonali, negli affari, nella politica, nei Tribunali

    Cinzia Gimelli

    2012-01-01

    Thanks to the scientist Paul Ekman we are now able to assess the type of influence that our non-verbal behaviour has on other people: we can now judge the ability of people to send non-verbal signals and know how to interpret them. We recognize that information provided by words in some cases is contradicted and refuted by the non-verbal expressions that accompany it, and that, when one cannot use words, information can be communicated through non-verbal signs. The emotions are states of mind...

  13. Gli archivi dell'Inquisizione in Italia: problemi storiografici e descrittivi. Roma, Archivio di Stato, 18 febbraio 2005

    Andrea Del Col (editor

    2006-01-01

    Full Text Available It includes the folowing essays:Andrea Del Col, Presentazione Adriano Prosperi, Nuove prospettive per una storia dell'Inquisizione Grado Giovanni Merlo, Problemi documentari dell'Inquisizione medievale in Italia Silvana Seidel Menchi, I giudici dell'Inquisizione romana: inquisitori e vescovi, commissari, nunzi, cardinali, papi Andrea Del Col, I documenti del Sant'Ufficio come fonti per la storia istituzionale e la storia degli inquisiti Marco Pizzo, La Stanza Storica dell'Archivio del Sant'Uffizio: un metodo di intervento Francesca Cavazzana Romanelli, Il censimento della documentazione inquisitoriale riguardante l'Italia Giovanna Paolin - Andrea Del Col, La banca dati dei processi e la ricostruzione completa dell'attività del Sant'Ufficio di Aquileia e Concordia

  14. La mente mente, il volto no. Gli indizi dell’inganno nei rapporti interpersonali, negli affari, nella politica, nei Tribunali

    Cinzia Gimelli

    2012-11-01

    Full Text Available Thanks to the scientist Paul Ekman we are now able to assess the type of influence that our non-verbal behaviour has on other people: we can now judge the ability of people to send non-verbal signals and know how to interpret them. We recognize that information provided by words in some cases is contradicted and refuted by the non-verbal expressions that accompany it, and that, when one cannot use words, information can be communicated through non-verbal signs. The emotions are states of mind associated with physiological and psychophysiological changes, with internal or external stimuli, or with natural or learned behaviour. They are associated with mood, temperament, personality and disposition, and motivation. In most cases, lies can be detected because of signs of hidden emotions; the more intense and varied the emotions are, the more likely it is that a lie is unintentionally betrayed. The facial expressions and body language can reveal deep emotions: the presence of a specific emotional state is indicated by a shaky voice, an alteration of tone, a sunny smile, or a furrowed brow. Unlike ordinary expressions and gestures, it is difficult to hide micro-expressions and micro-gestures. Micro-expressions express the seven universal emotions: disgust, anger, fear, sadness, happiness, surprise, and contempt. In 1990 Paul Ekman expanded this list of basic emotions, including a range of positive and negative emotions, not all encoded in facial muscles. These emotions are amusement, contempt, embarrassment, excitement, guilt, pride, relief, satisfaction, pleasure, and shame. Micro-gestures are hard to conceal too, because every little movement is like a word that has a different meaning according to the use made of it in a ‘sentence’. Therefore, in order to analyze a gesture correctly one should keep in mind the context in which it is expressed.

  15. 40 CFR 721.7260 - Polymer of poly-ethylene-polyamine and alkanediol di-gly-cidyl ether.

    2010-07-01

    ... 40 Protection of Environment 30 2010-07-01 2010-07-01 false Polymer of poly-ethylene-polyamine and... New Uses for Specific Chemical Substances § 721.7260 Polymer of poly-ethylene-polyamine and alkanediol... chemical substance identified generically as polymer of polyethylenepolyamine and alkanediol diglycidyl...

  16. L'incomprensione tra gli amanti nel poema Khosrow e Širin di Neúàmi

    Paola Orsatti

    2010-01-01

    The problem of a lawful wedding is a crucial point in the plot of the poem. Khosrow’s constant efforts to elude marriage to Širin are due to interference of the historical sources. Indeed Širin too is a historic character: she was Khosrow’s influential Christian wife. In NeÛÁmi’s poem, Širin’s Christian faith is concealed; but Khosrow’s resistance to marriage with her is probably an echo of the opposition that their union encountered in court circles and among the artistocracy, since Širin was a woman of humble origins, as well as being Christian. The way she defends her reputation and her good name can be seen as a reaction against the bad light in which she was portrayed in the historical sources. Neúàmi has skilfully made use of these sources in order to voice his concept of love, of passionate love, as a source of suffering and incomprehension between lovers but also as a cosmic force which permeates every aspect of life, and the most important experience in the poet’s life. At the same time he gives a thorough psychological analysis of the characters, unimaginable for those times.

  17. Ramón Franco Bahamonde: l’adesione al golpe, i rapporti con gli italiani e la morte

    Edoardo Grassia

    2013-10-01

    Full Text Available In the “second part of his life”, that is to say after his choice of abandoning the Republican front to support the 1936 pronunciamento, Ramón had become an enemy of his former allies and a friend of his former enemies. However as General Alfredo Kindelan said: «nobody shook hands with him nor wanted to have relations with him» among his old enemies. In the group of the newly formed allies there were also the Italian soldiers faithful to Mussolini, that the Spanish pilot wanted to kill only a few years earlier. This article first describes friends and enemies of the Caudillo de España’s brother, the takes into account some documents to give a clear indication of the bad relationships between Ramón Franco and the Italian armed forces, both in Spain and in occasion of his visits to Italy. Moreover, some reflections on the circumstances of the Spanish pilot’s death will be proposed.

  18. The gas turbine: Present technology and future developments; La turbina a gas: Tecnologie attuali e gli sviluppi futuri

    Minghetti, E [ENEA, Centro Ricerche Casaccia, Rome (Italy)

    1997-03-01

    The gas turbine is the most widely used prime mover all over the world for either power generation or mechanical drive applications. The above fact is due to the recent great improvements that have been done especially in terms of efficiency, availability and reliability. The future for gas turbine technological development looks very promising. In fact, although tremendous growth has already taken place, there is still the potential for dramatic improvements in performance. Compared with the competitive prime movers (conventional steam power plants and reciprocating piston engines) the gas turbine technology is younger and still following a strong growth curve. The coming decades will witness the continued increasing in turbine inlet temperature, the development of new materials and refrigeration systems and the commercialization of inter cooled system and steam cooled turbines. With the very soon introduction of the {sup G }and {sup H }technology, expected single and combined cycle efficiencies for heavy duty machines are respectively 40% and 60%, while maintaining single digit levels in pollutant emissions. In this report are given wide information on gas turbine present technology (Thermodynamics, features, design, performances, emission control, applications) and are discussed the main lines for the future developments. Finally are presented the research and technological development activities on gas turbine of Italian National Agency for new Technology Energy and the Environment Energy Department.

  19. Ma cosa fanno gli americani? Una breve indagine sui ruoli e i compiti dei landscape architect statunitensi (II

    Danilo Palazzo

    2015-11-01

    Full Text Available American landscape architecture firms seem to have retained between their competences that of city and urban design. To look out how real is this feeling, that refers to what is published on specialized journals and books, the paper display a reconstruction of phases, protagonists, and modalities that characterized the division between the two disciplines, in terms of separate professional associations and academic curriculum. The division appeared to be slow and viscous.The evaluation of city-design-oriented activities conducted by self-defined landscape architecture’s firms is developed since the recognition of the design activities undertook by some of the American firms awarded in 2004 by the ASLA, the American Society of Landscape Architect. 

  20. Thermodynamic properties of peptide solutions 20. Partial molar volumes and isothermal compressions for some tripeptides of sequence gly-X-gly (X = gly, ala, leu, asn, thr, and tyr) in aqueous solution at T = 298.15 K and p = (10–120) MPa

    Hedwig, Gavin R.; Høiland, Harald

    2016-01-01

    Highlights: • Sound speeds were measured for aqueous solutions of some tripeptides at high pressures. • Partial molar volumes and isothermal compressions were derived for T = 298.15 K and p = (10–120) MPa. • The partial molar volumes for non-polar amino acid side-chains decrease with increasing pressure. • The partial molar volumes for polar side-chains do not change significantly with increasing pressure. - Abstract: Sound speeds have been measured for aqueous solutions of six tripeptides of sequence glycyl-X-glycine, where X is one of the amino acids glycine, alanine, leucine, asparagine, threonine, and tyrosine at T = 298.15 K and at the pressures p = (10, 20, 40, 60, 80, 100, and 120) MPa. Using methods described in previous work, these sound speeds were used to derive the partial molar volumes at infinite dilution, V_2"o, the partial molar isentropic compressions at infinite dilution, K_S_,_2"o, and the partial molar isothermal compressions at infinite dilution, K"o_T_,_2 {K"o_T_,_2 = −(∂V_2"o/∂p)_T}, for the tripeptides in aqueous solution at the elevated pressures. The results were used to calculate the partial molar volumes and partial molar isothermal compressions for the various amino acid side-chains over the pressure range p = (10–120) MPa.

  1. Particle range in a laser-plasma generated soft X-ray chamber

    Bollanti, S.; Letardi, T. [ENEA, Centro Ricerche Frascati, Rome (Italy). Dipt. Innovazione; Zheng, C. [EL.EN, Calenzano, Florence (Italy)

    1999-07-01

    Some analytical forms are deduced for calculating the flight range of a spherical particle ejected from the laser plasma target and retarded by gas resistance. it is shown that the gas pressure influence on viscosity can not be neglected when are estimated the expansion ranges fro debris of various sizes in a helium gas-buffered, laser produced plasma chamber. [Italian] Vengono ricavate alcune formule analitiche per il calcolo del range di frammenti sferici espulsi con velocita' iniziale dati e frenati dalla resistenza di un fondo gassoso. Si mostra che nei gas considerati non si puo' ignorare influenza della pressione nella viscosita' del gas.

  2. Primary structure of the precursor for the sea anemone neuropeptide Antho-RFamide (less than Glu-Gly-Arg-Phe-NH2)

    Darmer, D; Schmutzler, C; Diekhoff, D

    1991-01-01

    Neuropeptides containing the carboxylterminal sequence Arg-Phe-NH2 are found throughout the animal kingdom and are important substances mediating neuronal communication. Here, we have cloned the cDNA coding for the precursor protein of the sea anemone neuropeptide (Antho-RFamide) less than Glu...... harbors four other putative neuropeptides that are much less related to Antho-RFamide. This report shows that the biosynthetic machinery for neuropeptides in coelenterates, the lowest animal group having a nervous system, is already very efficient and similar to that of higher invertebrates...

  3. Conformational changes in fragments D and double-D from human fibrin(ogen) upon binding the peptide ligand Gly-His-Arg-Pro-amide.

    Everse, S J; Spraggon, G; Veerapandian, L; Doolittle, R F

    1999-03-09

    The structure of fragment double-D from human fibrin has been solved in the presence and absence of the peptide ligands that simulate the two knobs exposed by the removal of fibrinopeptides A and B, respectively. All told, six crystal structures have been determined, three of which are reported here for the first time: namely, fragments D and double-D with the peptide GHRPam alone and double-D in the absence of any peptide ligand. Comparison of the structures has revealed a series of conformational changes that are brought about by the various knob-hole interactions. Of greatest interest is a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket. Additionally, in the absence of the peptide ligand GPRPam, GHRPam binds to the gamma-chain pocket, a new calcium-binding site being formed concomitantly.

  4. La copertura dei rischi finanziari nelle imprese non finanziarie italiane attraverso gli strumenti derivati (The Hedging of Financial Risks Using Derivatives by Italian Non-financial Firms

    Gianluca Bison

    2012-04-01

    Full Text Available The paper deals with the use of derivatives by Italian non-financial firms, in order to analyse existing theories of hedging behaviour, and provides empirical evidence on a potential differentiation of determinants of derivative use over time. Univariate and multivariate analyses show that the determinants of derivative use reveal marked differences when compared in the different years, so they may change over time. In general, we find that the most determinant variable is foreign sales, and analysis therefore suggests the hypothesis that exposure to exchange rate risk is the strongest determinant in derivative use. Interest rate risk exposure seems to be less relevant. Moreover, from 1997, another variable proves determinant, firm size, which suggests that economies of scale could also be a key variable in derivative use. With respect to financial distress cost models, tax and agency cost theories, the results are mixed.           JEL Codes: G11, G31, G32, F31Keywords: Financial Risk, Firm, Hedging        

  5. Postumano. Il corpo tra psicoanalisi, biopolitica, realtà virtuale e arte Istituto italiano per gli Studi Filosofici – Napoli, 27 maggio 2016

    LO SAPIO, LUCA

    2016-12-01

    Full Text Available Posthuman. The Body between Psychoanalysis, Biopolitics, virtual Reality and Art The posthuman has become a topic of central interest for the definition of certain key-elements of Western thought. The May 27, 2016 in Naples an interesting meeting has been held, in which various speakers discussed on the effects that post-human and posthumanism have been producing in art, philosophy, psychoanalysis and literature. In particular, it was shown that the posthuman is a complex issue that requires multiple strategies to be addressed and understood effectively. The posthuman evokes fears and hopes and in particular it is directly connected with the vocation of man to self-transcendence, a somehow natural tendency which is leading to a general reorganization of Western man’s identity structures and to a general reshaping of his profound imagery.

  6. Global Value Chain Indicators: Application to the Italian Sectors - Gli indicatori della global value chain: un’applicazione ai settori italiani

    Ricotta, Fernanda

    2010-01-01

    In this paper the extent of international fragmentation of production in Italian manufacturing industries for the years 1985, 1995 and 2000 is assessed with different indicators. The objective is to determine where fragmentation is most prevalent and to provide a description of the key characteristics of those sectors. The paper first presents a survey of global value chain indicators and how these have been used in the economic literature. The second part supplies empirical evidence on the g...

  7. Molecularly imprinted nanoparticles with recognition properties towards a laminin H-Tyr-Ile-Gly-Ser-Arg-OH sequence for tissue engineering applications

    Rosellini, Elisabetta; Barbani, Niccoletta; Giusti, Paolo; Ciardelli, Gianluca; Cristallini, Caterina

    2010-01-01

    Nanotechnology is an emerging field that promises to revolutionize medicine and is increasingly used in tissue engineering applications. Our research group proposed for the first time molecular imprinting as a new nanotechnology for the creation of advanced synthetic support structures for cell adhesion and proliferation. The aim of this work was the synthesis and characterization of molecularly imprinted polymers with recognition properties towards a laminin peptide sequence and their application as functionalization structures in the development of bioactive materials. Nanoparticles with an average diameter of 200 nm were synthesized by precipitation polymerization of methacrylic acid in the presence of the template molecule and trimethylpropane trimethacrylate as the cross-linking agent. The imprinted nanoparticles showed good performance in terms of recognition capacity and selectivity. The cytotoxicity tests showed normal vitality of C2C12 myoblasts cultured in the medium that was put in contact with the imprinted polymers. After the deposition on the polymeric film surface, the imprinted particles maintained their specific recognition and rebinding behaviour, showing an even higher quantitative binding than free nanoparticles. Preliminary in vitro cell culture tests demonstrated the ability of functionalized materials to promote cell adhesion, proliferation and differentiation, suggesting that molecular imprinting can be used as an innovative functionalization technique.

  8. L’Italia compie 150 anni. E gli italiani? Una riflessione sul senso di appartenenza, sull’identità e sull’insuperata asimmetria tra Italia e italiani

    Riccardo Giumelli

    2011-11-01

    Full Text Available If Italy in 2011 becomes 150 years old, than how old are Italians? So to paraphrase the title of this paper, we introduce mischievously the idea that Italians have a different age. Obviously, we will try to respond, reflecting on the sense of belonging to a collective identity, in our case the Italian one, in times of globalization or, better, glocalization. The reflexion about Italian identity, or Italic as we are going to call it, comes from the extraordinary and special change due to the global processes that deconstruct the socio-political systems of nation-state created by the Treaty of Westphalia in 1648, which ended the Thirty Years' War. In conclusion, what interests us is the cultural identity is increasingly characterized by processes of human, objects and symbols mobility that no longer recognize, as they did for a long time, territorial barriers and boundaries.

  9. Comparison of human IgE-binding soya bean allergenic protein Gly m I with the antigenicity profiles of calf anti-soya protein sera

    Hessing, M.; Bleeker, H.; Tsuji, H.; Ogawa, T.; Vlooswijk, R.A.A.

    1996-01-01

    In the food, particularly the feed, industry, large quantities of soya bean protein products are used for the formulation of end-products destined for human or animal consumption. These basic ingredients and the final end products often have to fulfill specific requirements regarding the presence of

  10. [A protective effect of GLY272SER polymorphism of GNB3 gene in development of essential hypertension and its relations with environmental hypertension risk factors].

    Polonikov, A V; Solodilova, M A; Ivanov, V P; Shestakov, A M; Ushachev, D V; Vialykh, E K; Vasil'eva, O V; Poliakova, N V; Antsupov, V V; Kabanina, V A; Kupriianova, Ia S; Bulgakova, I V; Kozhukhov, M A; Tevs, D S

    2011-01-01

    To study associations of C825T (rs5443) and G272S (rs16932941) polymorphisms of GNB3 gene in Russian population of the Central Chernozem region with essential hypertension (EH) risk; to elicit the role of environmental risk factors in realization of EH predisposition in this gene genotypes carriers. We studied DNA samples obtained from 205 EH patients and 207 healthy individuals. EH patients were treated in Kursk hospitals. Genotyping of GNB3 gene polymorphisms was conducted by polymerase chain reaction and restriction analysis. Prevalence of 82ST allele of GNB3 gene in EH patients and healthy individual was 0.334 and 0.295, respectively, of 272S allele--0.037 and 0.058, respectively. We found no significant differences by prevalence of genotypes of gene GNB3 polymorphisms C825T and G272S in EH patients and healthy individuals. Non-smoking carriers of 272GS genotype had a low risk of EH (OR 0.42 in 95% CI from 0.18 to 0.97; p = 0.04). Smokers had no protective effect of this genotype. The protective effect of 272GS genotype was also found in individuals with low or moderate alcohol drinking habits (OR 0.29 in 95% CI from 0.11 to 0.77, p = 0.02) and in individuals without chronic exposure to stress (OR 0.29 in 95% CI from 0.09 to 0.91, p = 0.04). In contrast, hard drinkers and patients exposed to chronic stress had no protective effect of heterozygous genotype 272GS of gene GNB3. G272S polymorphism of GNB3 gene can be considered as a new genetic marker of predisposition to EH. The protective effect depends of environmental factors associated with high risk to develop EH.

  11. L'Energy Amplifier di Rubbia : gli studi per la produzione di un'energia nucleare pulita ed il loro impatto sul piano economico

    Sessano, D; Panizza, R

    2000-01-01

    The growing interest towards new energy forms is a common feature of many different studying fields: physics, engineering, economics and politics. The main reason is that human population is growing fast (there's a prevision of about nine billion people in 2050) and with it is increasing the energy request, both for the industrialized world and for the developing countries. This means that the actual organization of the energy market will not be able to satisfy the needs of the near future: clean energy for everybody. The International Energy Agency provided a "business as usual" scenario where is shown that by the year 2030 the world energy demand will be 70% higher than now and the electricity demand would follow the same pattern, needing somewhat like 3475 GW of new electric power. The logic conclusion is that we'll need all the energy forms we know: fossil fuels, nuclear and renewable energy. Nowadays the energy - and particularly the electricity - market is facing large problems: global warming and depen...

  12. «We have ceased to be a nation in retreat». Margaret Thatcher e gli usi strategici del conflitto

    Lidia De Michelis

    2015-03-01

    Full Text Available This essay explores Margaret Thatcher’s political agency and leadership style in terms of her reliance on discursive and media strategies which thrived on the “confusion” of gender stereotypes and her ability to play with multiple, long-established archetypes of women in power, characterized by violent and aggressive femininity. Thatcher’s frequent self-projection as a “masculine” character, the “honorary man” of British politics, and her special grip on British public opinion and fantasies, were never disjointed from her ability to don, exploit, subvert and redirect “motherly’ (or at least “womanly” performances at will, in order to serve her political agenda. Focusing on her strategic deployment of a rhetoric and discourses of aggression, divisiveness, conflict and war, the essay highlights Thatcher’s ability to re-create and keep alive the idea of a national community always under siege, where extraordinary economic, disciplinary and defence measure were required in order to resist and overcome domestic and foreign “enemies”. The Falklands War will be taken as case study, for it provides an outstanding example of Thatcher’s powerful revision of female political authority against the backdrop of the conventionally masculine, heroic imaginary of war. Her masterly and coterminous performances as war leader and “super-mother” of the nation allowed for a complex and hybrid self-projection which still acts as an empowering model whose disciplinary subtext entails a nostalgic investment in imperial national history.

  13. "In questo", il teatro gli scenari della commedia dell'arte "In this", the theater and the scenery of the commedia dell'arte

    Roberto Cuppone

    2001-01-01

    Full Text Available Il segreto della commedia dell'arte è stato nel paradosso di Maschera e Improvvisazione, Tipo Fisso e Testo Variabile, Conservazione e Innovazione. Per questo è stato nei secoli esaltata o denigrata: considerata reazionaria dalla Rivoluzione francese e esaltata come rivoluzionaria dai Romantici. Nel Novecento, Copeau, Mejerchol'd, Mnouchkine e Fo hanno dovuto fare i conti con questa ambivalenza. Dunque ciò che oggi si chiama commedia dell'arte è "reazionario" o "rivoluzionario"? "Popolare"o "populista"?The secret of the Commedia dell'Arte lies in the paradox between Mask and Improvisation, Fixed Character and Variable Text, Conservation and Innovation. Therefore along the centuries it has been exalted or vilified: considered as reactionary during the French revolution and praised by the romantics. In the 20th century Copeau, Meierhold, Mnouchkine and Fo faced this ambivalence. Is that which is today called commedia dell'arte "reactionary" or "revolutionary"? "Popular" or "populist"?

  14. Gli italo-somali dell’Amministrazione Fiduciaria Italiana della Somalia (AFIS: una memoria dimenticata tra le pagine dell’Italia postcoloniale

    Michele Pandolfo

    2017-07-01

    Full Text Available At the end of the Second World War the United Nations gave to Italy a special protectorate on the own former colony by the International Trusteeship System of Somalia, which concludes in 1960. During the Fifties there are many relationships between the italian men and the somali women: from these liaisons there are a lot of children which have different destinies and their history represents one of the heredity of the italian past in Africa. From this italo-somali métissage, rises an association who asks the recognition of all the pains suffered by a part of that community and in the same time it asks a collective reflections of the historic memories about the Trusteeship System.

  15. Nuove strategie di disseminazione e figure emergenti: gli innovation brokers + Beyond dissemination of research findings: innovation brokers as emerging figures in stimulating agricultural innovation

    Klerkx, L.W.A.

    2012-01-01

    More and more it is recognised that innovation cannot be explained by a linear approach to innovation in which public sector agricultural research and extension delivers new technology in a pipeline configuration through a dissemination approach, but calls for systems approach in which innovation is

  16. Investigating the role of GXXXG motifs in helical folding and self-association of plasticins, Gly/Leu-rich antimicrobial peptides.

    Carlier, Ludovic; Joanne, Pierre; Khemtémourian, Lucie; Lacombe, Claire; Nicolas, Pierre; El Amri, Chahrazade; Lequin, Olivier

    2015-01-01

    Plasticins (PTC) are dermaseptin-related antimicrobial peptides characterized by a large number of leucine and glycine residues arranged in GXXXG motifs that are often described to promote helix association within biological membranes. We report the structure and interaction properties of two plasticins, PTC-B1 from Phyllomedusa bicolor and a cationic analog of PTC-DA1 from Pachymedusa dacnicolor, which exhibit membrane-lytic activities on a broad range of microorganisms. Despite a high number of glycine, CD and NMR spectroscopy show that the two plasticins adopt mainly alpha-helical conformations in a wide variety of environments such as trifluoroethanol, detergent micelles and lipid vesicles. In DPC and SDS, plasticins adopt well-defined helices that lie parallel to the micelle surface, all glycine residues being located on the solvent-exposed face. Spectroscopic data and cross-linking experiments indicate that the GXXXG repeats in these amphipathic helices do not provide a strong oligomerization interface, suggesting a different role from GXXXG motifs found in transmembrane helices. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Statistical sources and methodology of elaboration for the regional, national and European energy indicators; Fonti statistiche e metodologie di elaborazione per gli indicatori energetici regionali, nazionali ed europei

    Bianco, R; Perrella, G [ENEA, Centro Ricerche Casaccia, Rome (Italy). Dip. Energia

    1996-10-01

    This report is aimed to presenting the energy-economic and structural data system called EIS (energy indicators system), about the statistical sources and the different methodologies of data valuation and indicators elaboration. This system, elaborated in the activity of ERG-PROM Division of ENEA (Italian Agency for New Technologies, Energy and the Environment), is aimed to analyze the energy by quantity and quality use, in the different sectors, for each final aim and for every source. In the report are defined all the procedure and unit of measure used to process the data bank, so the reader can to interpret the right meaning of the variables and indicators and can make further process.

  18. Somatic mosaicism for the COL7A1 mutation p.Gly2034Arg in the unaffected mother of a patient with dystrophic epidermolysis bullosa pruriginosa

    van den Akker, P. C.; Pasmooij, A. M. G.; Meijer, R.; Scheffer, H.; Jonkman, M. F.

    Dystrophic epidermolysis bullosa (DEB) is a heritable blistering disorder caused by mutations in the type VII collagen gene, COL7A1. Although revertant mosaicism is well known in DEB, 'forward' somatic mosaicism, in which a pathogenic mutation arises on a wild-type (WT) background, extending beyond

  19. Somatic mosaicism for the COL7A1 mutation p.Gly2034Arg in the unaffected mother of a patient with dystrophic epidermolysis bullosa pruriginosa

    Akker, P.C. van den; Pasmooij, A.M.; Meijer, R.; Scheffer, H.; Jonkman, M.F.

    2015-01-01

    Dystrophic epidermolysis bullosa (DEB) is a heritable blistering disorder caused by mutations in the type VII collagen gene, COL7A1. Although revertant mosaicism is well known in DEB, 'forward' somatic mosaicism, in which a pathogenic mutation arises on a wild-type (WT) background, extending beyond

  20. Comprendere gli elementi intangibili dell’esperienza turistica heritage: una ricerca netnografica / Understanding the intangible elements of the heritage-based tourism experience: a netnographic study

    Giacomo Marzi

    2016-06-01

    The aim of the study is to investigate how intangible elements of cultural heritage influence visitors’ perception of a particular tourist destination. This study uses the qualitative research method of netnography, examining travel diaries from online communities in order to analyze the recorded perceptions of heterogeneous visitors of four different tourist destinations (Florence, Naples, Rome and Venice. Evidences show a high quantity of references related to local food and visited communities (everyday life, contact with locals, etc. in the examined diary texts. Such a result indicates how these elements remain anchored in the mind of tourists and, therefore, may be interpreted as significant factors in the evaluation of holiday experiences. Regarding the practical implications, this study aims to improve the decision makers’ awareness about the importance of intangible heritage elements, specifically concerning heritage tourism.

  1. «Fatta l’Europa bisogna fare gli europei». Considerazioni su una educazione e una formazione all’Europa

    Merete Amann Gainotti

    2009-05-01

    Full Text Available This essay faces an important argument: the building of European citizenship. The Author makes a historical reconstruction on this argument and then she writes about the immagine of Europe on scholastic books. What do childreen know about European Union and on their role of European citizens?

  2. Lorenzo Lotto e gli strumenti del mestiere: la periferia come consapevole scelta strategica / Lorenzo Lotto and the tools of the trade: the periphery as a conscious strategic choice

    David Frapiccini

    2014-06-01

    Full Text Available Con il testamento del 25 marzo 1531 Lorenzo Lotto stabiliva la ripartizione dei modelli in gesso e in cera e dei propri disegni tra tre suoi antichi allievi: il bergamasco Francesco Bonetti, il veneziano Pietro di Giovanni – in quel momento attivo a Ragusa, l’odierna Dubrovnik – e il marchigiano maestro Giulio da Amandola, identificabile con Giulio Vergari. Dalle provenienze dei tre artefici si manifesta una chiara mappa geografica, come se Lotto avesse consapevolmente deciso di lasciare i ferri del mestiere nei luoghi della sua esperienza pittorica: a Bergamo, a Venezia e nelle Marche. Contemporaneamente Lotto frequenta i pittori Alessandro Oliverio, Bonifacio de’ Pitati da Verona e Girolamo da Santa Croce: il secondo, stando alle disposizioni testamentarie, avrebbe dovuto completare le opere rimaste incompiute alla morte di Lotto, mentre al terzo sarebbero andati alcuni strumenti della professione a beneficio dei propri figli. Si viene così a delineare un gruppo di artisti marginale rispetto al contesto lagunare, ma orientato verso committenze di entroterra o di area adriatica. La consapevolezza acquisita da Lotto nell’incidenza territoriale della sua attività spiega la ragione delle reminiscenze iconografiche e formali bergamasche in parte della produzione risalente agli estremi anni marchigiani (1549-1556, ovvero a quella fase inaugurata con l’Assunzione anconetana di San Francesco alle Scale, pala realizzata nel 1549-1550 con la collaborazione di Giuseppe Belli da Ponteranica. With the will of March 25, 1531 Lorenzo Lotto established the distribution of models in plaster and wax and their designs to three of his former pupils: Francesco Bonetti from Bergamo, the Venetian Pietro di Giovanni – at that time active in Ragusa, today's Dubrovnik – and maestro Giulio from Amandola identified with Giulio Vergari. The origins of the three painters manifest a clear geographical map, as if Lotto had consciously decided to leave the tools of the trade in the places of his painting experience: in Bergamo, Venice and the Marches. At the same time Lotto associated with the painters Alessandro Oliverio, Bonifacio de' Pitati from Verona and Girolamo da Santa Croce: the second, according to the testamentary dispositions, would have to complete the work left unfinished at the death of Lotto, while the third would go some tools of the profession for the benefit their children apprentices. It is thus to define a group of artists marginal compared to the Venetian milieu, but addressed toward patrons of inland or Adriatic area. The knowledge acquired by Lotto in the territorial incidence of his work explains the reason for his reminiscences of iconographic and formal pattern of the times of Bergamo in the production dating back to the last years in the Marches (1549-1556, particularly at that stage of the Assumption of San Francesco alle Scale to Ancona, altarpiece carried out in 1549-1550 in collaboration with Giuseppe Belli from Ponteranica.

  3. Familial amyloidotic polyneuropathy with severe renal involvement in association with transthyretin Gly47Glu in Dutch, British and American-Finnish families

    Haagsma, EB; Hawkins, PN; Benson, MD; Lachmann, HJ; Bybee, A; Hazenberg, BPC

    Familial amyloidotic polyneuropathy (FAP) is an autosomal dominant disorder associated with more than 80 different transthyretin (TTR) mutations. The clinical features of FAP are broad and variable, but knowledge of the pattern and natural history of disease associated with particular mutations

  4. The Gly16 allele of the G16R single nucleotide polymorphism in the β2-adrenergic receptor gene augments the glycemic response to adrenaline in humans

    Rokamp, Kim Z.; Staalsø, Jonatan M.; Zaar, Morten

    2017-01-01

    Cerebral non-oxidative carbohydrate consumption may be driven by a ß2-adrenergic mechanism. This study tested whether the 46G > A (G16R) single nucleotide polymorphism of the ß2-adrenergic receptor gene (ADRB2) influences the metabolic and cerebrovascular responses to administration of adrenaline....... Forty healthy Caucasian men were included from a group of genotyped individuals. Cardio- and cerebrovascular variables at baseline and during a 60-min adrenaline infusion (0.06 μg kg-1 min-1) were measured by Model flow, near-infrared spectroscopy and transcranial Doppler sonography. Blood samples were...... obtained from an artery and a retrograde catheter in the right internal jugular vein. The ADRB2 G16R variation had no effect on baseline arterial glucose, but during adrenaline infusion plasma glucose was up to 1.2 mM (CI95: 0.36-2.1, P

  5. ‹‹IN AIUTO ALL’UNITÀ DELLA LINGUA E CONTRO GLI ERRORI PROVENIENTI DAL DIALETTO››. IL SILLABARIO E IL VOCABOLARIETTO DI ANTONINO TRAINA

    Michela Dota

    2016-02-01

    Full Text Available Il titolo del presente contributo condensa da una parte le coordinate operative della legislazione scolastica elementare post-unitaria, in particolare riguardo all’ortoepia, e d’altra parte la ricezione di quelle coordinate nella pubblicistica scolastica elementare coeva. Il saggio considera entrambi i cardini della questione: dopo aver esposto brevemente le linee guida della legislazione circa il rapporto tra lingua e dialetti e sulla corretta pronuncia, si analizzano due strumenti didattici, concepiti per la duplice utenza dei maestri e degli alunni, nei quali si concretano due atteggiamenti sensibilmente diversi sulla questione ortoepica. Infine, dall’analisi di questi due sussidi si dedurrà il rapporto, raccomandato più o meno esplicitamente, che doveva intercorrere tra dialetto e lingua nelle aule scolastiche. «In help to the unit of language and against the mistakes coming from dialects». Spelling book and little vocabulary by Antonino TrainaThe title of this contribution epitomizes the praxis of the post-unification legislation concerning primary school, particularly about orthoepy, and the reception of that legislation by the authors of textbooksfor primary school. After having exposed the guidelines of the legislation about the relation between Italian and dialects and about correct pronunciation, this article analyzes two teaching tools (a spelling book and a vocabulary, that can be used by both teachers and students. It’s notable that the spelling book and the vocabulary have a different perspective on orthoepy. Finally, from their analysis it is deduced the model proposed for the relation between Italian and dialects in classrooms.

  6. Preliminary results for filtering-ecosystems in Val Trebbia (PC - Italy); Primi risultati per gli ecosistemi-filtro della Val Trebbia (PC)

    Malcevschi, Sergio [Pavia, Univ. (Italy). Dipt. di Biologia Animale; Bisogni, G Luca [N.Q.A. s.r.l., Pavia (Italy); Bernini, Fabrizio [Amministrazione Provinciale di Piacenza, Piacenza (Italy)

    1997-09-01

    Some artificial wetlands were constructed in 1993-1994 in the Trebbia Valley (Piacenza, Italy). These plants worked as further treatment of secondary effluents of five little towns in the valley: Bobbio, Travo, Perino-Dolgo, Marsaglia, Ottone. The project and the realization considered the systems as Filtering Ecosystems and not a simple Aquatic Plant Systems. In 1994-1995 a research by University of Pavia was carried out, on the filtering ecosystem of Bobbio. Removal efficiency was 9,9 for phaecal coliforms, and 65% for nitrates.

  7. Gli investimenti di portafoglio nelle economie di mercato emergenti:tendenze, dimensioni e problemi (Portfolio Investment in Emerging Market Economies: Trends, Dimensions and Issues

    Dilip K. Das

    2012-04-01

    Full Text Available Securitised financial flows to emerging market economies have become an important feature of the global capital flows. The principal focus of this paper is on an in-depth analysis of current trends in securitised financial flows. It lays special emphasis on private portfolio equity investment into the emerging market economies. The paper begins with the analysis of the process of stimulation of these flows, and identifies the institutional, structural and non-cyclical factors behind them. One of the points it emphasises is the progressively important role of institutional investors, which is the causal factor behind a significant increase in the quantum of portfolio investment into the emerging market economies. These flows were adversely affected by the financial crises of the 1990s. The subject matter of this paper also includes two of the most important policy issues, namely, the "hot money and cold money" issue and the volatility issue.       JEL Codes: G11, O16, F32, P33Keywords: Capital Flows, Financial Flows, Portfolio

  8. Gli investimenti di portafoglio nelle economie di mercato emergenti:tendenze, dimensioni e problemi (Portfolio Investment in Emerging Market Economies: Trends, Dimensions and Issues

    Dilip K. Das

    2000-09-01

    Full Text Available Securitised financial flows to emerging market economies have become an important feature of the global capital flows. The principal focus of this paper is on an in-depth analysis of current trends in securitised financial flows. It lays special emphasis on private portfolio equity investment into the emerging market economies. The paper begins with the analysis of the process of stimulation of these flows, and identifies the institutional, structural and non-cyclical factors behind them. One of the points it emphasises is the progressively important role of institutional investors, which is the causal factor behind a significant increase in the quantum of portfolio investment into the emerging market economies. These flows were adversely affected by the financial crises of the 1990s. The subject matter of this paper also includes two of the most important policy issues, namely, the "hot money and cold money" issue and the volatility issue.

  9. Il clima dell'areale del Lago Maggiore durante gli ultimi cinquant'anni (The climate of Lago Maggiore area during the last fifty years)

    Ambrosetti, Walter; Barbanti, Luigi; Rolla, Angelo

    2006-01-01

    Data collected in over 50 years' activity at the C.N.R. meteorological station of Pallanza were analysed to see whether any changes have taken place in some climatic elements and to quantify the extent of such changes. A statistical analysis of the respective historical series was performed on nine meteorological parameters (solar radiation, insolation, air temperature, atmospheric pressure, humidity, cloud cover, precipitation, evaporation and wind), and on two other strictly limnological fe...

  10. Gli esperimenti di “utopie concrete” con gli indigeni dell’America nel XVI secolo: Vasco de Quiroga e Bartolomé de Las Casas - The experiments of “concrete utopias” with the indigenous of America in the XVI century: Vasco de Quiroga and Bartolomé de Las Casas

    Antonino Colajanni

    2012-10-01

    Full Text Available This essay begins with some considerations about the diffusion in Europe, during the XVIth and XVIIth centuries, of many philosophical-literary writings imagining some perfect societies located in spaces and times very far from the actual environments. Those writings represent a series of radical and irreducible criticism to the characters of contemporary societies. Thomas More, the first and most important representative of the “utopian thinking”, affected with his celebrated work Utopia (1516 the philosophical and political ideas of the period. His fundamental book influenced also directly the intellectual formation as well as the activities and the writings of some ecclesiastic figures of the time engaged in the corrections and modification of the European presence in the Americas (missionaries, theologians, and reformers of the strategies of evangelization, who dedicated years to the concrete realization of some ideas and projects related to utopian thinking. This essay thoroughly examines two important figures of the time: Vasco de Quiroga, Bishop of Michoacán (Mexico, and the well known Dominican theologian and jurist Bartolomé de Las Casas (in the second part of his life Bishop of Chiapas, great adversary of the Spanish Conquest of Latin America. In different ways, and more or less inspired by utopian thinking, these missionary authorities renowned as “defenders of the Indians” proposed and partly realized some projects of radical transformation of the entire life (material and spiritual as well of the indigenous populations of some regions of Latin America. The author proposes to make a sharp distinction between Quiroga and Las Casas. He thinks that they possibly could best be defined as “social reformers” inspired by utopian thinking rather than as “concrete utopians”.

  11. Biological properties of adriamycin bound to biodegradable polymeric carriers

    Hoes, C.J.T.; Hoes, C.J.T.; Grootoonk, J.; Grootoonk, J.; Duncan, R.; Hume, I.C.; Bhakoo, M.; Bouma, J.M.W.; Feijen, Jan

    1993-01-01

    Three different conjugates having adriamycin (ADR) bound to the side chain carboxyl groups of high-molecular weight poly (¿--glutamic acid) (PGA) either directly or by interpolation of GlyGly and GlyGlyGlyLeu spacers, respectively, were compared with respect to immunogenicity and cytotoxicity in

  12. BIOLOGICAL PROPERTIES OF ADRIAMYCIN BOUND TO BIODEGRADABLE POLYMERIC CARRIERS

    HOES, CJT; GROOTOONK, J; DUNCAN, R; HUME, IC; BHAKOO, M; BOUMA, JMW; FEIJEN, J

    Three different conjugates having adriamycin (ADR) bound to the side chain carboxyl groups of high-molecular weight poly(alpha-L-glutamic acid) (PGA) either directly or by interpolation of GlyGly and GlyGlyGlyLeu spacers, respectively, were compared with respect to immunogenicity and cytotoxicity in

  13. RESEARCH ARTICLE

    women with PCOS are at a higher risk for gestational diabetes, miscarriages, ..... meta- analysis on PCOS women reported that Gly/Arg and Gly/Gly genotype is ... IRS1 and IRS2 polymorphisms influence glucose homeostasis and could.

  14. L'inglese per gli italiani; un corso programmato autodidattico realizzato dalla Britannica. Laboratorio Linguistico Individuale Anglotutor [English for Italians; A Self-Instructional Programmed Course developed by Encyclopaedia Britannica].

    Encyclopaedia Britannica, Inc., Chicago, IL.

    This eight-volume audiolingual course in programmed English for Italian students appears with instructions and grammatical explanations in Italian and numerous illustrations. Special attention is given to contrastive pronunciation problems. The materials, intended for use on the beginning level, are accompanied by tapes. (AMM)

  15. Formare gli psicologi dell’emergenza all’intervento psicologico in ricerca di dispersi e al sostegno delle famiglie di persone scomparse1. L’esperienza del laboratorio al 5° Campo Scuola di Marco di Rovereto

    Rina Maria Galeaz

    2012-03-01

    Full Text Available On the occasion of the 5th training camp for the volunteer psychologists of the Civil Protection, it was proposed the workshop “Psychological interventions in search and rescue operations”, managed by experts in support to the aid networks. The aim was to integrate, for the first time, the topic of psychological support to relatives and search and rescue personnel with that of missing persons' families. This experience allowed to develop some reflections about emergency psychologists' training and the intervention models for the dissemination of good practices and the sharing of know-hows with other emergency professionals.

  16. Chiudo la porta e m' arrabio! Gli effetti interpersonali delle emozioni nelle trattative pubbliche e private [I close the door and get angry! The interpersonal effects of emotions in public and private negotiations

    Pietroni, D.; van Kleef, G.A.; Steinel, W.; Rumiati, R.

    2008-01-01

    Negotiators' emotions strongly affect counterparts' behavioral reactions. In particular, some research show that, compared to happy negotiators, angry negotiators get higher concessions, whereas other research indicate that happiness pays more than anger. This incongruence has been spelled out

  17. Fenomeni d’interferenza nell’apprendimento dell’italiano da parte di parlanti spagnolo. Un’indagine a partire da un test a scelta multipla per gli studenti dell’Università Complutense di Madrid.

    Francesco Zurlo

    2009-12-01

    Full Text Available Normal 0 14 MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Tabella normale"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:10.0pt; font-family:"Times New Roman";} È noto che nell’apprendimento di lingue affini come italiano e spagnolo, il fenomeno del transfer linguistico gioca spesso un ruolo fondamentale. La percezione di vicinanza e di congruenza tra i due sistemi linguistici è alla base del frequente trasferimento da parte degli apprendenti, di regole, espressioni ed abitudini proprie della loro L1 nella L2, che non inficiando in molti casi il successo comunicativo rischiano alla lunga di cristallizzarsi e fossilizzarsi. Nella ricerca, svolta a partire da un test a scelta multipla proposto ad alcuni alunni dei corsi di italiano dell’Università Complutense di Madrid, si rilevano e descrivono alcune delle tipologie più ricorrenti di errori dovuti ad interferenza della L1, mettendo in rilievo la loro incidenza e la loro influenza nel determinare la specificità del percorso di apprendimento dell’italiano da parte degli ispanofoni, contraddistinto da accelerazioni e rallentamenti, arresti e stabilizzazioni.  Normal 0 14 MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Tabella normale"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:10.0pt; font-family:"Times New Roman";} It is known that when learning similar languages, such as Italian and Spanish, the phenomenon of linguistic transfer often plays a fundamental role.  The perception of the two language systems as close and congruent is at the basis of the learners’ frequent transfer of rules, expressions and habits belonging to their L1 into the L2  which, since in many cases their communicative success is not hindered, risk taking root and becoming fossilized in the long term. The survey, based on a multiple-choice test run with students on an Italian course at Madrid’s Università Complutense, identifies and describes some of the most recurrent types of errors due to interference from L1, highlighting their incidence and influence in determining the specific nature of the learning route for Spanish speakers studying Italian, marked by acceleration and slowdowns, halts and moments of stabilization.     

  18. A Gly65Val substitution in an actin, GhACT_LI1, disrupts cell polarity and membrane anchoring of F-actin resulting in dwarf, lintless Li1 cotton plants

    • Actin polymerizes to form the cytoskeleton and organize polar growth in all eukaryotic cells. Species with numerous actin genes are especially useful for the dissection of actin molecular function due to redundancy and neofunctionalization. Here, we investigated the role of a cotton (Gossypium hi...

  19. Sentiment Analysis per analizzare gli effetti del cinema sulla percezione dei luoghi. Il caso pugliese / Sentiment Analysis to study the effects of cinema on the perception of places. The case of Puglia Region

    Valentina Albanese

    2016-05-01

    Full Text Available Il film induced iourism è un fenomeno ormai indiscutibile e che va affrontato con sistematicità e metodologie sempre più raffinate per consentire ai policy makers di sfruttarne più consapevolmente le potenzialità. In Puglia la realizzazione di pellicole di successo ha provocato effetti turistici e territoriali notevoli. Per poter comprendere realmente se e quanto l’incoming turistico pugliese sia influenzato, nella sua dimensione quantitativa e qualitativa, dall’immagine veicolata dal cinema, si ipotizza qui l’utilizzo di una nuova metodologia di analisi: la Sentiment Analysis. Si intende passare al setaccio i Big Data tematici, tramite una scansione intelligente dei social network e poi riportare le valutazioni (sentiment sul territorio pugliese espresse nei diversi luoghi virtuali di conversazione da parte della domanda turistica. Questa tipologia di studio del dato è del tutto innovativa per il settore cineturistico e può portare ad esiti del tutto inattesi sovvertendo in alcuni casi le interpretazioni prettamente soggettive dei dati quantitativi più tradizionali. Film induced tourism is an undeniable phenomenon and it is necessary to study it sistematically and with sophisticated methods to allow policy makers to exploit, more consciously, its potentiality. In Apulia the successful movies maybe have caused tourism and territorial remarkable effects. In order to understand if and how incoming in Apulia is influenced, in quantitative and qualitative terms, from the image conveyed by cinema, we’ll use a new method of analysis: the Sentiment Analysis. It means making an intelligent scanning of social networks and then bring feedback (sentiment about Apulia expressed in different virtual places of conversation by the tourist demand. This kind of opinion mining study is totally innovative for film induced tourism and can lead to outcomes completely unexpected subverting, in some cases, subjective interpretations of the quantitative, more traditional, data.

  20. Characterization of rPEPT2-mediated Gly-Sar transport parameters in the rat kidney proximal tubule cell line SKPT-0193 cl.2 cultured in basic growth media

    Bravo, Silvina A; Nielsen, Carsten Uhd; Frokjaer, Sven

    2005-01-01

    The rat proximal kidney tubule cell line SKPT-0193 cl.2 (SKPT) expresses the di-/tripeptide transporter PEPT2 (rPEPT2) and has been used to study PEPT2-mediated transport. Traditionally, SKPT cells have been cultured in growth media supplemented with epidermal growth factor (EGF), apotransferrin,...

  1. In margine a un convegno. Gli intellettuali/scrittori ebrei e il dovere della testimonianza. Per Giorgio Bassani, “di là dal cuore” (Firenze, 7-8-9 novembre 2016

    Martina Romanelli

    2016-12-01

    Full Text Available This brief article aims to investigate some aspects of an international conference held last November in Florence. In the year devoted to Giorgio Bassani, it was an occasion to address aspects of Jewish cultural data, with special reference to the reactions of writers in general and an individual in particular to the unspeakable horrors of the Holocaust. The conference acknowledged the need to face up to harsh reality and search for an adequate historiographical, ethical and literary response.

  2. Individuare gli incendiari. Fermare i piromani. Una professione / Identify the arsonists. Detain the pyromaniacs. A profession / Identifier les incendiaires. Arrêter les pyromanes. Un métier

    Giuseppe Vadalà

    2008-04-01

    Full Text Available From the end of the Second World War onwards, at an international level we witness an increasing development of the so called environmental emergency. The environment has become that immaterial resource that must assure all citizens well-being and renewable basic life resources for the coming generation, but globalization has produced a remarkable growth of negative results resulting in new types of menace.Forest fires directly represent one of these threats causing the destruction of both forest patrimony and natural ecosystems worldwide, contributing to an increase in the carbon dioxide level in the atmosphere. The Act n. 353/2000 concerning forest fires reorganized the whole sector. The aim of the law is to control and to fight the risk of forest fires thanks to the implementation of a synergic system.The criminal system of sanctions represents another type of enforcing action in order to protect woods from fires, but it can not be the only solution to this problem assuming that we have to face a very complex and multifaceted event which causes should be to identify any widespread illegal phenomena or any extremely superficial behaviour as well as in criminal ones, which take on specific characteristics when occurring in a rural context. The identification of a solution for this intricate problem could be research, both at a preventive and at a control level, into activities able to operate on the many different causes that are at the origin of that behaviour which is the cause of forest fires and arson.Depuis la fin de la seconde guerre mondiale, la prétendue émergence des questions environnementales au niveau international s'est développée de façon exponentielle. L'environnement est ainsi devenu ce bien immatériel qui doit assurer aux citoyens la salubrité et les ressources vitales renouvelables pour les générations futures. Toutefois, la mondialisation a provoqué une forte augmentation des effets dangereux pour l'environnement et cela a créé des typologies nouvelles de menaces.Les incendies de forêts représentent exactement une de ces menaces parce qu'ils causent non seulement la destruction du patrimoine forestier mondial, mais aussi celle des écosystèmes naturels et ils contribuent à l'augmentation du niveau de l'anhydride carbonique dans l'atmosphère.La loi cadre en matière d'incendies de forêts, n° 353 du 21 novembre 2000, a réorganisé tout le secteur. Cette norme a pour but de contrôler et de s'opposer à la menace des incendies de forêts par la mise en place d'un système cohérent et synergique. Le système de sanctions représente un moyen supplémentaire de renforcer la défense de forêts contre les incendies, mais il n'est pas décisif en soi car il faut faire face à un événement complexe et multiforme, dont les causes sont liées à l'illégalité répandue, aux comportements superficiels et/ou criminels qui revêtent des caractéristiques spécifiques dans le milieu rural.La résolution d'une problématique si complexe implique une lutte à la fois préventive et répressive grâce aux actions résolues contre les nombreuses causes à la base des comportements d'incendies volontaires et involontaires.

  3. Gli effetti della scala mobile sull'inflazione e sulle politiche di stabilizzazione: un commento al modello Modigliani-Padoa-Schioppa (The effects of wage indexation on inflation and on stabilization policies: a comment on the Modigliani-Padoa Schioppa

    G. VITALETTI

    2014-03-01

    Full Text Available In a recent article, F. Modigliani and T. Padoa-Schioppa argued that indexing at 100% with frequent wage adjustments rises the general rate of inflation associated with a given gap between the price level desired by businesses and that compatible with the real bargaining wage of workers. The present work compares this examination of an economy with wages indexed at 100% with an alternative situation in which indexing is absent or not effective in protecting wages. It is shown that in the presence of expectations of inflation incorporated into the determination of wage rates: 1 if there is a gap between the price level desired by businesses and that compatible with the real wage desired by workers, inflation is growing rather than stable as in the M.-P.S. model; 2 the "return" of inflation through restrictive maneuvers is accomplished with greater difficulty and involves greater time and costs than in the M.-P.S. model. JEL: E24, E31, J31

  4. P. Giulierini, P. Bruschetti, F. Gaultier, L. Haumesser, Gli Etruschi dall’Arno al Tevere, Le collezioni del Louvre a Cortona

    Joncheray, Claire

    2018-01-01

    Dans le contexte d’un passage de la valorisation du patrimoine culturel de l’État italien à la municipalité de Cortone (Province d’Arezzo), le Museo dell’Accademia Etrusca e della città di Cortona (ci-après MAEC) s’investit dans une politique de collaboration internationale : il proposait en 2008 une exposition sur les chefs-d’œuvre d’art étrusque conservés au musée de Saint-Pétersbourg, et il propose en 2011 d’accueillir ceux du Louvre au Palazzo Cas...

  5. Quando gli investimenti rappresentano un vincolo. Contributo alla discussione sulla crisi italiana nella crisi internazionale.(When investments represent a constraint. A note on the discussion on Italy’s crisis within the international crisis

    Stefano Lucarelli

    2013-07-01

    Full Text Available The article continues the debate on the "crisis within the crisis" that characterises Italy's economy. The authors argue that while the international and specifically the European crisis greatly contributed to the deterioration of Italy's economic prospects, the country is affected by longer term issues of its own. To analyse such problems, the article focuses on the growing inability of the Italian private sector to innovate. It does so by analysing in a comparative perspective private sector expenditure for fixed investments and for research and development. The policy conclusions are surprising in so far as a surge of investments would seriously risk to negatively contribute to Italy's current account, thus further aggravating.    

  6. Preparation and characterization of a hetero functional system of gold nanoparticles labeled with {sup 99m}Tc and conjugated to the sequence Arg-Gly-Asp for detection in vivo of angio genesis and evaluation of their toxicity in Hyalella aztec; Preparacion y caracterizacion de un sistema heterofuncional de nanoparticulas de oro marcadas con Tecnecio-99m y conjugadas a la secuencia Arg-Gly-Asp para la deteccion in vivo de angiogenesis y la evaluacion de su toxicidad en Hyalella azteca

    Morales A, E.

    2012-07-01

    Integrin s play critical roles in many physiological processes including angio genesis and also contribute to pathological events such as tumor invasion and metastasis. The {alpha}{sub v}{beta}{sub 3} integrin is expressed in normal endothelial cells but it is over-expressed in the tumor neo vasculature. Peptides based on the Arginine-Glycine-Aspartic acid (RGD) sequence have been reported as molecules with high affinity and selectivity for the {alpha}{sub v}{beta}{sub 3} integrin. Recent studies have demonstrated that conjugating peptides to gold nanoparticles (AuNP) produces biocompatible and stable multifunctional systems with target-specific molecular recognition due to multivalent effects produced by multiple simultaneous interactions between peptides and their receptors. The first aim of this research was to prepare a m ultimeric system of {sup 99m}Tc labeled gold particles conjugated to c[RGDfK(C)] and to evaluate its biological behavior as a potential radiopharmaceutical for molecular imaging of {alpha}{sub v}{beta}{sub 3} tumor expression. Hidrazinonicotinamide-G GC (HYNIC-G GC) and C[RGDfK(C)] peptides were synthesized and conjugated to AuNP (20 nm) by means of spontaneous reaction of the thiol groups of cysteine. The nano conjugate was characterized by transmission electron microscopy, Fourier transform-infrared, Ultraviolet-vis, X-ray photoelectron spectroscopy and Raman spectroscopy. To obtain {sup 99m}Tc-HYNIC-G GC-AuNP-c[RGDfK(C)], the {sup 99m}Tc-HYNIC-G GC radio peptide was first prepared and added to the AuNP solution followed by c[RGDfK(C)]. Radiochemical purity (Rp) was determined by size-exclusion HPLC and I TLC-Sg analyses. In vitro binding studies were carried out in {alpha}{sub v}{beta}{sub 3} receptor-positive C6 glioma cancer cells. Biodistribution studies were accomplished in athymic mice with C6-induced tumors with blocked and non blocked receptors, and images were obtained using a micro-SPECT/CT. Transmission electron microscopy and spectroscopy techniques demonstrated that AuNP were functionalized with peptides. Rp was 96 {+-} 2% without post-labeling purification. {sup 99m}Tc-HYNIC-G GC-AuNP-RGD showed specific recognition for {alpha}{sub v}{beta}{sub 3} integrin s expressed in C6 cells, and 3 h after i.p. administration in mice, the tumor uptake was 8.18 {+-} 0.57% Id/g. Micro-SPECT/CT images showed evident tumor uptake. {sup 99m}Tc-HYNIC-G GC-AuNP-RGD demonstrates properties suitable for use as a new target-specific agent for molecular imaging of tumor {alpha}{sub v}{beta}{sub 3} expression. the second aim of this research was to determine the eco toxicological risk, measured by oxidative stress induction in Hyalella aztec, of the well-characterized multifunctional RGD-AuNP system (Tc-HYNIC-G GC-AuNP-c[RGDfK(C)]). In acute toxicological studies, a median lethal concentration of 1.83 cm{sup 2} per milliliter of medium was established for the Tc-HYNIC-G GC-AuNP-c[RGDfK(C)] conjugate. Tests assessing the effects of oxidative stress such as lipo peroxidation and protein carbonyl protein content suggest that this nano conjugate does not induces changes in oxidative markers because the activity of antioxidant defenses including catalase, glutathione peroxidase and superoxide dismutase do not show an increase related to oxidative stress imbalance. Due to the chain length and steric effect of the peptides attached to gold nanoparticles by Au-S bonds, the Tc-HYNIC-G GC-AuNP-C[RGDfK(C)] complex does not react with intracellular glutathione or thiol-proteins and, therefore, does not induce increases in reactive oxygen species. Therefore, Tc-HYNIC-G GC-AuNP-c[RGDfK(C)] (20 nm) is a new chemically stable diagnostic pharmaceutical that does not induce eco toxicity measured by the oxidative stress induction in Hyalella aztec after complete radionuclide decay. This study suggests that Hyalella aztec is a suitable model animal for environmental toxicology studies of nanoparticles. (Author)

  7. Note critiche sui rapporti tra diritto ed economia

    Tuzet, Giovanni

    2017-01-01

    Il saggio inquadra teoricamente i principali rapporti fra diritto ed economia e analizza i vari metodi e approcci dell'analisi economica del diritto, rilvandone gli aspetti critici e discutendo infine gli approcci comportamentali e la "nudge theory".

  8. Applications of Pharmacogenetics in Revealing Variations in ...

    acer

    taking place in molecular biology techniques. Drug action is now more ... the latest findings of genetic variations in pharmacological targets related to disorders of major .... Similarly, Gly 9 allele. (Ser9Gly) of the dopamine D3 receptor gene.

  9. Localization of glycine-containing neurons in the Macaca monkey retina

    Hendrickson, A.E.; Koontz, M.A.; Pourcho, R.G.; Sarthy, P.V.; Goebel, D.J.

    1988-01-01

    Autoradiography following 3H-glycine (Gly) uptake and immunocytochemistry with a Gly-specific antiserum were used to identify neurons in Macaca monkey retina that contain a high level of this neurotransmitter. High-affinity uptake of Gly was shown to be sodium dependent whereas release of both endogenous and accumulated Gly was calcium dependent. Neurons labeling for Gly included 40-46% of the amacrine cells and nearly 40% of the bipolars. Synaptic labeling was seen throughout the inner plexiform layer (IPL) but with a preferential distribution in the inner half. Bands of labeled puncta occurred in S2, S4, and S5. Both light and postembedding electron microscopic (EM) immunocytochemistry identified different types of amacrine and bipolar cell bodies and their synaptic terminals. The most heavily labeled Gly+ cell bodies typically were amacrine cells having a single, thick, basal dendrite extending deep into the IPL and, at the EM level, electron-dense cytoplasm and prominent nuclear infoldings. This cell type may be homologous with the Gly2 cell in human retina and the AII/Gly2 of cat retina. Gly+ amacrines synapse most frequently onto Gly- amacrines and both Gly- and Gly+ bipolars. Gly+ bipolar cells appeared to be cone bipolars because their labeled dendrites could be traced only to cone pedicles. The pattern of these labeled dendritic trees indicated that both diffuse and midget types of biopolars were Gly+. The EM distribution of labeled synapses showed Gly+ amacrine synapses throughout the IPL, but these composed only 11-23% of the amacrine population. Most of the Gly+ bipolar terminals were in the inner IPL, where 70% of all bipolar terminals were labeled

  10. Teoria della relatività

    Pauli, Wolfgang

    1958-01-01

    Fondamenti della teoria della relatività ristretta ; ausili matematici ; gli sviluppi della teoria della relatività ristretta concernenti la cinematica ; gli sviluppi della teoria della relatività ristretta concernenti l'elettrodinamica ; gli sviluppi della teoria della relatività ristretta concernenti la meccanica e la dinamica generale ; gli sviluppi della teoria della relatività ristretta concernenti la termodinamica e la statistica ; teoria della relatività generale ; teorie sulla natura delle particelle elementari elettricamente cariche.

  11. Determination of the torsion angles of alanine and glycine residues of model compounds of spider silk (AGG){sub 10} using solid-state NMR methods

    Ashida, Jun; Ohgo, Kosuke; Komatsu, Kohei; Kubota, Ayumi; Asakura, Tetsuo [Tokyo University of Agriculture and Technology, Department of Biotechnology (Japan)], E-mail: asakura@cc.tuat.ac.jp

    2003-02-15

    Spiders synthesize several kinds of silk fibers. In the primary structure of spider silk, one of the major ampullate (dragline, frame) silks, spidroin 1, and flagelliform silk (core fibers of adhesive spiral), there are common repeated X-Gly-Gly (X = Ala, Leu, Pro, Tyr, Glu, and Arg) sequences, which are considered to be related to the elastic character of these fibers. In this paper, two dimensional spin diffusion solid-state NMR under off magic angle spinning (OMAS), {sup 13}C chemical shift contour plots, and Rotational Echo DOuble Resonance (REDOR) were applied to determine the torsion angles of one Ala and two kinds of Gly residues in the Ala-Gly-Gly sequence of {sup 13}C=O isotope-labeled (Ala-Gly-Gly){sub 10}. The torsion angles were determined to be ({phi}, {psi}) = (-90 deg., 150 deg.) within an experimental error of {+-}10 deg. for each residue. This conformation is characterized as 3{sub 1} helix which is in agreement with the structure proposed from the X-ray powder diffraction pattern of poly(Ala-Gly-Gly). The 3{sub 1} helix of (Ala-Gly-Gly){sub 10} does not change by formic acid treatment although (Ala-Gly){sub 15} easily changes from the silk I conformation (the structure of Bombyx mori silk fibroin before spinning in the solid state) to silk II conformation (the structure of the silk fiber after spinning) by such treatment. Thus, the 3{sub 1} helix conformation of (Ala-Gly-Gly){sub 10} is considered very stable. Furthermore, the torsion angles of the 16th Leu residue of (Leu-Gly-Gly){sub 10} were also determined as ({phi}, {psi}) = (-90 deg., 150 deg.) and this peptide is also considered to take 3{sub 1} helix conformation.

  12. Journal of Chemical Sciences | Indian Academy of Sciences

    The kinetics of interaction between cis-[RuCl(Me2SO)3(H2O)2]+ and some selected dipeptides such as glycyl glycine(gly gly), glycyl alanine(Gly-L-ala) and glycyl-L-leucine(gly leu) has been studied spectrophotometrically as a function of [RuCl(Me2SO)3(H2O) 2 + ], [dipeptide] and temperature at a particular pH(5.0), where ...

  13. High efficient removal of chromium (VI) using glycine doped polypyrrole adsorbent from aqueous solution

    Ballav, N

    2012-08-01

    Full Text Available Glycine doped polypyrrole (PPy-gly) adsorbent was prepared via in situ polymerization of pyrrole (Py) monomer in the presence of glycine (gly) for the removal of Cr(VI). Formation of PPy homopolymer and inclusion of gly in the PPy matrix were...

  14. Effects of β2-adrenergic receptor polymorphisms on asthma severity ...

    EL-HAKIM

    -ray ... Moreover, statistical analysis revealed association of Arg/Arg ... However, no difference in distribution of Arg/Gly .... age and gender. .... A meta-analysis including a total of 28 studies ... concluded that, Gly/Gly had a much higher risk for.

  15. Genetic markers, which define the occurrence and course of bronchial asthma in children

    Banadyha N.V.

    2016-03-01

    Full Text Available Purpose: to analyze the frequency of polymorphic loci associations rs 1042713 (Arg16Gly of ADRβ2 gene in children with bronchial asthma. Patients and methods: in-depth clinical examination using the special methods of investigation, conducted to 62 children suffering from bronchial asthma. The results of investigation. As a result of depth collection of anamnesis, it was revealed that in 73.68% of patients the anamnesis was unburdened. Among the examined patients, bronchial asthma manifested at the early age in 18 children (33.96% in preschool age in 17 children (32.08%, and in a primary school in 18 children (33.96%. The early debut of disease associated with genotype Arg16Gly, while late manifestation observed in children with genotype Gly16Gly. Mostly the family inheritance depends on mother health, regardless of the severity of bronchial asthma. Іt was found that in both types of inheritance (paternal and maternal dominated genotype Arg16Gly. Found that girls often associated with asthma genotype Gly16Gly (56.52% and Arg16Gly (39.13% while the boys with genotype Arg16Gly (53.84%, less with Gly16Gly (38.89%. However, genotype Arg16Arg was observed in individual patients and in the case of intermittent disease. In case of allergen-induced and virus-induced phenotypes the genotype Arg16Gly was more often diagnosed. It was clarified that intermittent flow associated with two genotypes: Arg16Gly (47.37% and Gly16Gly (42.11%. The persistent mild course of bronchial asthma replied to genotype Gly16Gly (64.71%, but with moderate persistent — to Arg16Gly (57.69%. A good bronchodilator response was observed in patients with genotype diagnosed Arg16Gly and Gly16Gly. At the same time, patients with genotype Arg16Arg ADRβ2 needed the use of combined drugs to overcome the attack. Conclusions: Allelic polymorphism differences of ADRβ2 gene in children with asthma were diagnosed and it indicates that dependence of debut was genetically based as well

  16. Glycine transporter dimers: evidence for occurrence in the plasma membrane.

    Bartholomäus, Ingo; Milan-Lobo, Laura; Nicke, Annette; Dutertre, Sébastien; Hastrup, Hanne; Jha, Alok; Gether, Ulrik; Sitte, Harald H; Betz, Heinrich; Eulenburg, Volker

    2008-04-18

    Different Na(+)/Cl(-)-dependent neurotransmitter transporters of the SLC6a family have been shown to form dimers or oligomers in both intracellular compartments and at the cell surface. In contrast, the glycine transporters (GlyTs) GlyT1 and -2 have been reported to exist as monomers in the plasma membrane based on hydrodynamic and native gel electrophoretic studies. Here, we used cysteine substitution and oxidative cross-linking to show that of GlyT1 and GlyT2 also form dimeric complexes within the plasma membrane. GlyT oligomerization at the cell surface was confirmed for both GlyT1 and GlyT2 by fluorescence resonance energy transfer microscopy. Endoglycosidase treatment and surface biotinylation further revealed that complex-glycosylated GlyTs form dimers located at the cell surface. Furthermore, substitution of tryptophan 469 of GlyT2 by an arginine generated a transporter deficient in dimerization that was retained intracellulary. Based on these results and GlyT structures modeled by using the crystal structure of the bacterial homolog LeuT(Aa), as a template, residues located within the extracellular loop 3 and at the beginning of transmembrane domain 6 are proposed to contribute to the dimerization interface of GlyTs.

  17. Recombinant Brucella abortus gene expressing immunogenic protein

    Mayfield, J.E.; Tabatabai, L.B.

    1991-06-11

    This patent describes a synthetic recombinant DNA molecule containing a DNA sequence. It comprises a gene of Brucella abortus encoding an immunogenic protein having a molecular weight of approximately 31,000 daltons as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis under denaturing conditions, the protein having an isoelectric point around 4.9, and containing a twenty-five amino acid sequence from its amino terminal end consisting of Gln-Ala-Pro-Thr-Phe-Phe-Arg-Ile-Gly-Thr-Gly-Gly-Thr-Ala-Gly-Thr-Tyr-Tyr-Pro-Ile-Gly-Gly-Leu-Ile-Ala, wherein Gln, Ala, Pro, Thr, Phe, Arg, Ile, Gly, Tyr, and Leu, respectively, represent glutamine, alanine, proline, threonine, phenylalanine, arginine, isolecuine, glycine, tyrosine, and leucine.

  18. Effect of substituting barley with glycerol as energy feed on feed intake, milk production and milk quality in dairy cows in mid or late lactation

    Gaillard, Charlotte; Sørensen, Martin Tang; Vestergaard, Mogens

    2018-01-01

    intake, and milking frequency were recorded daily, while milk composition and milk FA daily were analyzed weekly. Milk sensory analysis was performed on fresh and 7 d stored samples for the four diets. The PMR intake increased almost 1 kg from Gly0 to Gly12, and decreased by approximately 1 kg from Gly12......The experiment reported in this research paper aimed to determine the level at which glycerol can substitute barley in grass-clover silage-based ration for dairy cows in mid or late lactation, without affecting milk production, milk composition, milk free fatty acid (FFA) profile, and milk sensory...... quality. Forty Holstein cows were used in a 4 × 4 Latin square experimental design. Crude glycerol substituted barley in the partially mixed ration (PMR) of the cows at inclusion levels of 0% (Gly0), 6% (Gly6), 12% (Gly12), and 18% (Gly18) of dietary dry matter (DM). Individual milk production, feed...

  19. Glycine transporter dimers: evidence for occurrence in the plasma membrane

    Bartholomäus, Ingo; Milan-Lobo, Laura; Nicke, Annette

    2008-01-01

    membrane based on hydrodynamic and native gel electrophoretic studies. Here, we used cysteine substitution and oxidative cross-linking to show that of GlyT1 and GlyT2 also form dimeric complexes within the plasma membrane. GlyT oligomerization at the cell surface was confirmed for both GlyT1 and GlyT2......Different Na(+)/Cl(-)-dependent neurotransmitter transporters of the SLC6a family have been shown to form dimers or oligomers in both intracellular compartments and at the cell surface. In contrast, the glycine transporters (GlyTs) GlyT1 and -2 have been reported to exist as monomers in the plasma...

  20. Effect of arginine:lysine and glycine:methionine intake ratios on dyslipidemia and selected biomarkers implicated in cardiovascular disease: A study with hypercholesterolemic rats.

    Venkatesh, Ravula; Srinivasan, Krishnapura; Singh, Sridevi Annapurna

    2017-07-01

    The effect of intake ratios of arginine (Arg): lysine (Lys) and glycine (Gly): methionine (Met) on lipid profile and selected cardiovascular disease markers, was studied, in rats maintained on a hypercholesterolemic diet. The rise in blood cholesterol was countered by 32%, 24%, and 49%, respectively, through increased oral supplementation of Arg, Gly, and Arg+Gly; a corresponding increase in plasma phospholipids at the end of the 8-week study was observed. The elevated plasma cholesterol to phospholipids ratio was countered by 27, 40, and 57%, respectively, through oral supplementation of Arg, Gly, and Arg+Gly. The elevation in hepatic cholesterol was lowered by 18, 29, and 51%, respectively, while phospholipids concentration was concomitantly increased by these amino acids. The elevated cholesterol to phospholipids ratio was, thus, significantly countered in the hypercholesterolemic situation by orally supplemented Arg, Gly, and Arg+Gly. Increased plasma asymmetric dimethylarginine (ADMA) levels, under hypercholesterolemic conditions, were lowered by 12, 15 and 34%, respectively, while plasma symmetric dimethylarginine (SDMA) levels were lowered by 14, 10 and 17%, respectively, with orally supplemented Arg, Gly and Arg+Gly. Only Gly and Arg+Gly decreased plasma homocysteine levels. Total nitric oxide (NO) concentration was considerably increased by Gly supplementation in hypercholesterolemic rats. Thus, altered ratios of Arg:Lys or Gly:Met offered beneficial influence on the lipid profile and plasma levels of ADMA, SDMA and homocysteine in hypercholesterolemic rats. Optimal beneficial effects, among ratios tested, was observed when Arg:Lys and Gly:Met ratios were maintained in ratios of 1:1 and 2:1, respectively. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  1. Egidio Calzini (1857-1928 e gli studi di storia dell'arte in Romagna e nelle Marche tra XIX e XX secolo / Egidio Calzini (1857-1928 and the studies of art history in Romagna and Marche regions between the 19th and the 20th centuries

    Francesco De Carolis

    2016-06-01

    The paper aims to discover the career and the critical profile of Egidio Calzini, an art historian who actively contributed to the study of the artistic heritage in Romagna and Marches between 19th and 20th century. Following a diachronic order of his studies, the research underlines both his relationship with the rest of scholars of that time, examining unpublished sources, and his best discoveries, analyzing his publications. From the monography on Marco Palmezzano (1894 to direction of the specialized journal titled «Rassegna bibliografica dell’arte italiana» (1898-1916, the article defines his personality, who contributes to the period of the art history in which started the current features of the discipline, both about conservation and  research studies.

  2. GlyEFSA NDA Panel (EFSA Panel on Dietetic Product s, Nutrition and Allergies), 20 13 . Scientific Opinion on the substantiation of a health claim related to glycaemic carbohydrates and recovery of normal muscle function (contraction) after stren uous exercise pursuant to Article 13(5) of Regulation

    Tetens, Inge

    Following an application from Aptonia, submitted for authorisation of a health claim pursuant to Article 13(5) of Regulation (EC) No 1924/2006 via the Competent Authority of France, the EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA) was asked to deliver an opinion on the scientific...... substantiation of a health claim related to glycaemic carbohydrates and recovery of normal muscle function (contraction) after strenuous exercise. The food constituent, glycaemic carbohydrates, which is the subject of the health claim, is sufficiently characterised in relation to the claimed effect. Recovery...... to the recovery of normal skeletal muscle function (contraction), the Panel concludes that a cause and effect relationship has been established between the consumption of glycaemic carbohydrates and recovery of normal muscle function (contraction) after strenuous exercise....

  3. La differenza tra noi e “gli altri”: su alcune mostre coloniali milanesi organizzate nel Ventennio fascista / The difference between us and “the others”: notes on some colonial exhibitions in Milan during the “Ventennio” (the Fascist period

    Silvia Colombo

    2016-12-01

    In this essay, the results obtained by the protagonists who worked for these shows – artists, architects, customers... – will be also considered, together with the didactic intentions illustrated by the team of organisers. In that way it will be possible to underline the communication instruments used by the regime in order to impress the public, to understand the key-role played by the exhibitions during the “Ventennio”; and, finally, to perceive the xenophobic perspective employed by the PNF to educate the masses in seeing “the others”.

  4. Definition of the Bacillus subtilis PurR operator using genetic and bioinformatic tools and expansion of the PurR regulon with glyA, guaC, pbuG, xpt-pbuX, yqhZ-folD, and pbuO

    Saxild, Hans Henrik; Brunstedt, K.; Nielsen, K.I.

    2001-01-01

    The expression of the pur operon, which encodes enzymes of the purine biosynthetic pathway in Bacillus subtilis, is subject to control by the purR gene product (PurR) and phosphoribosylpyrophosphate. This control is also exerted on the purA and purR genes. A consensus sequence for the binding...... of PurR, named the PurBox, has been suggested (M. Kilstrup, S.G. Jessing, S.B. Wichmand-Jorgensen, M. Madsen, and D. Nilsson, J. Bacteriol. 180:3900-3906, 1998). To determine whether the expression of other genes might be regulated by PurR, we performed a search for PurBox sequences in the B. subtilis...

  5. L’influenza del cinema d’animazione nella promozione (cineturistica delle destinazioni. Gli effetti del fenomeno Frozen sul turismo norvegese / The influence of animation cinema in the (cinetouristic promotion of destinations. The effects of the Frozen phenomenon on Norwegian tourism

    Enrico Nicosia

    2016-05-01

    Full Text Available Oggi le location sono diventate vere e proprie  destinazioni turistiche,  cioè  luoghi che offrono un insieme di relazioni, prodotti, servizi, elementi naturali ed artificiali, capaci di attrarre un certo numero di visitatori e soddisfarne le attese sia dal punto di vista funzionale che da quello emozionale.  Il Film Induced Tourism è una tendenza che si è affermata negli ultimi anni con la creazione di agenzie turistiche specializzate nel proporre tour guidati alle mete in cui siano state girate famose scene cinematografiche. Secondo diverse statistiche effettuate sul tema del Film Induced Tourism è dimostrato che, anche se è un fenomeno recente, è in crescita nel mercato turistico mondiale e si impone sempre più all’attenzione dei media, dei sociologi e degli operatori turistici. Con questo lavoro si cercherà di analizzare il Film Induced Tourism con l’intento di approcciarsi ad una nuova frontiera: il Cartoon Induced Tourism (il turismo indotto dalla visione di film d’animazione e cartoons, perché non solo la visione di un film o di una serie tv può indurre il potenziale (cineturista a viaggiare. Anche un film d’animazione, che per definizione è pura creazione, può suscitare allo spettatore le stesse emozioni di una pellicola girata in una location “reale” e spingerlo ad esplorare i luoghi da cui i registi hanno preso spunto per costruire o meglio disegnare il set in cui sono ambientati.  Nowadays locations are becoming tourist destinations, offering relations such as products, services, natural and manmade elements, able to attract a certain number of visitors, satisfying expectations from a functional point of view as well as from an emotional one. Film Induced Tourism represent a trend developed in the last years according with the creation of tourist agencies, specialised in guided tour to specific destination in which famous cinematographic scenes have been made. According to various statistics on Film Induced Tourism it was shown that, even if representing a recent phenomenon, it is a growing trend in the world tourism market, also attracting the  attention of media, sociologist and tourism field operator. With this work I try to analyze the Film Induced Tourism with the intention to approach to a new frontier: the Cartoon Induced Tourism (tourism-induced vision of animated films and cartoons, because not only watching a movie or of a TV series can induce potential (cine tourist to travel. Even an animated film, in this case Frozen, can arouse the viewer the same feeling of a film shot on location “real” and push to explore the places from which the directors have been inspired to build or rather draw the set in which they are set.

  6. Individuazione degli errori di scheggiatura nei diversi metodi di débitage del Paleolitico inferiore e medio. Gli esempi di Cà Belvedere di Monte Poggiolo (FC), Guado San Nicola 1 (IS), Riparo Tagliente (VR), Payre e Abri du Maras (Ardéche, Francia).

    Buonsanto, Cecilia

    2012-01-01

    The understanding of the management of natural resources by prehistoric man through an analysis of stone tools is useful to create parameters for the study of the level of manual dexterity and subsistence strategies in the Paleolithic. The study of knapping errors in experimental and archaeological lithic collections can reveal the presence of beginners or of experienced knappers and offers a comprehensive overview on the various errors associated with different methods of knapping débitage. ...

  7. Renewable energy sources and their contribution to gross domestic energy consumption in Italy in years 1992 and 1993; Censimento per il territorio italiano dei dati relativi alle fonti rinnovabili di energia ed al loro contributo al bilancio energetico per gli anni 1992 e 1993

    Menna, P [ENEA, Centro Ricerche, Portici (Italy). Dip. Energia

    1995-11-01

    In 1992, the gross domestic energy consumption in Italy was 167.5 MTOE (including bunkers) while in 1993 it decreased to 165.9 MTOE. The overall electricity demand passed from 244.8 TWh in 1992 to 246.6 TWh in 1993. In the same period, the overall contribution from Renewable Energy Sources (RES) kept almost constant at 5.y consumption. The RES contribution represented a share larger than gross National production of energy in 1992.

  8. Preparation and characterization of a hetero functional system of gold nanoparticles labeled with 99mTc and conjugated to the sequence Arg-Gly-Asp for detection in vivo of angio genesis and evaluation of their toxicity in Hyalella aztec

    Morales A, E.

    2012-01-01

    Integrin s play critical roles in many physiological processes including angio genesis and also contribute to pathological events such as tumor invasion and metastasis. The α v β 3 integrin is expressed in normal endothelial cells but it is over-expressed in the tumor neo vasculature. Peptides based on the Arginine-Glycine-Aspartic acid (RGD) sequence have been reported as molecules with high affinity and selectivity for the α v β 3 integrin. Recent studies have demonstrated that conjugating peptides to gold nanoparticles (AuNP) produces biocompatible and stable multifunctional systems with target-specific molecular recognition due to multivalent effects produced by multiple simultaneous interactions between peptides and their receptors. The first aim of this research was to prepare a m ultimeric system of 99m Tc labeled gold particles conjugated to c[RGDfK(C)] and to evaluate its biological behavior as a potential radiopharmaceutical for molecular imaging of α v β 3 tumor expression. Hidrazinonicotinamide-G GC (HYNIC-G GC) and C[RGDfK(C)] peptides were synthesized and conjugated to AuNP (20 nm) by means of spontaneous reaction of the thiol groups of cysteine. The nano conjugate was characterized by transmission electron microscopy, Fourier transform-infrared, Ultraviolet-vis, X-ray photoelectron spectroscopy and Raman spectroscopy. To obtain 99m Tc-HYNIC-G GC-AuNP-c[RGDfK(C)], the 99m Tc-HYNIC-G GC radio peptide was first prepared and added to the AuNP solution followed by c[RGDfK(C)]. Radiochemical purity (Rp) was determined by size-exclusion HPLC and I TLC-Sg analyses. In vitro binding studies were carried out in α v β 3 receptor-positive C6 glioma cancer cells. Biodistribution studies were accomplished in athymic mice with C6-induced tumors with blocked and non blocked receptors, and images were obtained using a micro-SPECT/CT. Transmission electron microscopy and spectroscopy techniques demonstrated that AuNP were functionalized with peptides. Rp was 96 ± 2% without post-labeling purification. 99m Tc-HYNIC-G GC-AuNP-RGD showed specific recognition for α v β 3 integrin s expressed in C6 cells, and 3 h after i.p. administration in mice, the tumor uptake was 8.18 ± 0.57% Id/g. Micro-SPECT/CT images showed evident tumor uptake. 99m Tc-HYNIC-G GC-AuNP-RGD demonstrates properties suitable for use as a new target-specific agent for molecular imaging of tumor α v β 3 expression. the second aim of this research was to determine the eco toxicological risk, measured by oxidative stress induction in Hyalella aztec, of the well-characterized multifunctional RGD-AuNP system (Tc-HYNIC-G GC-AuNP-c[RGDfK(C)]). In acute toxicological studies, a median lethal concentration of 1.83 cm 2 per milliliter of medium was established for the Tc-HYNIC-G GC-AuNP-c[RGDfK(C)] conjugate. Tests assessing the effects of oxidative stress such as lipo peroxidation and protein carbonyl protein content suggest that this nano conjugate does not induces changes in oxidative markers because the activity of antioxidant defenses including catalase, glutathione peroxidase and superoxide dismutase do not show an increase related to oxidative stress imbalance. Due to the chain length and steric effect of the peptides attached to gold nanoparticles by Au-S bonds, the Tc-HYNIC-G GC-AuNP-C[RGDfK(C)] complex does not react with intracellular glutathione or thiol-proteins and, therefore, does not induce increases in reactive oxygen species. Therefore, Tc-HYNIC-G GC-AuNP-c[RGDfK(C)] (20 nm) is a new chemically stable diagnostic pharmaceutical that does not induce eco toxicity measured by the oxidative stress induction in Hyalella aztec after complete radionuclide decay. This study suggests that Hyalella aztec is a suitable model animal for environmental toxicology studies of nanoparticles. (Author)

  9. Impact of the β-1 adrenergic receptor polymorphism on tolerability and efficacy of bisoprolol therapy in Korean heart failure patients: association between β adrenergic receptor polymorphism and bisoprolol therapy in heart failure (ABBA) study.

    Lee, Hae-Young; Chung, Wook-Jin; Jeon, Hui-Kyung; Seo, Hong-Seog; Choi, Dong-Ju; Jeon, Eun-Seok; Kim, Jae-Joong; Shin, Joon Han; Kang, Seok-Min; Lim, Sung Cil; Baek, Sang-Hong

    2016-03-01

    We evaluated the association between coding region variants of adrenergic receptor genes and therapeutic effect in patients with congestive heart failure (CHF). One hundred patients with stable CHF (left ventricular ejection fraction [LVEF] adrenergic receptor gene (ADRB1), the observed minor Gly allele frequency (Gly389Arg + Gly389Gly) was 0.21, and no deviation from Hardy-Weinberg equilibrium was observed in the genotypic distribution of Arg389Gly (p = 0.75). Heart rate was reduced from 80.8 ± 14.3 to 70.0 ± 15.0 beats per minute (p < 0.0001). There was no significant difference in final heart rate across genotypes. However, the Arg389Arg genotype group required significantly more bisoprolol compared to the Gly389X (Gly389Arg + Gly389Gly) group (5.26 ± 2.62 mg vs. 3.96 ± 2.05 mg, p = 0.022). There were no significant differences in LVEF changes or remodeling between two groups. Also, changes in exercise capacity and brain natriuretic peptide level were not significant. However, interestingly, there was a two-fold higher rate of readmission (21.2% vs. 10.0%, p = 0.162) and one CHF-related death in the Arg389Arg group. The ADRB1 Gly389X genotype showed greater response to bisoprolol than the Arg389Arg genotype, suggesting the potential of individually tailoring β-blocker therapy according to genotype.

  10. Effects of glycerin concentration in steam-flaked corn-based diets with supplemental yellow grease on performance and carcass characteristics of finishing beef steers.

    Buttrey, E K; Luebbe, M K; McCollum, F T; Cole, N A; MacDonald, J C; Hales, K E

    2015-07-01

    Forty-eight individually fed crossbred steers (initial BW = 381 ± 7.61 kg) were used to determine the effects of glycerin (GLY) concentration in steam-flaked corn (SFC)-based diets with added yellow grease on animal performance and carcass characteristics. Glycerin was included at 0, 2.5, 5, and 10% dietary DM replacing SFC. A completely randomized design was used with steers as the experimental unit, and the model included the fixed effects of diet. Contrast statements were used to determine linear and quadratic effects of GLY inclusion. Final BW and DMI were not affected (P > 0.27) by GLY concentration. Average daily gain and G:F based on live BW did not differ as GLY level increased in the diet from 0 to 10% of DM (P > 0.33). Carcass-adjusted final BW and carcass-adjusted G:F were also not affected by GLY concentration (P > 0.22); however, carcass-adjusted ADG tended to respond quadratically by decreasing from 2.5 to 5% GLY inclusion and increasing thereafter (P = 0.10). Calculated dietary NEm and NEg did not differ as GLY increased in the diet (P > 0.37). Hot carcass weight tended to respond quadratically, decreasing from 2.5 to 5% GLY and increasing thereafter (P = 0.10). Likewise, dressing percentage tended to respond quadratically by decreasing from 2.5 to 5% GLY inclusion and increasing to 10% GLY inclusion (P = 0.09). Fat thickness decreased linearly as GLY inclusion increased in the diet (P 0.21). Calculated yield grade (YG) decreased linearly as GLY increased in the diet from 0 to 10% of DM (P = 0.04). Based on our results, there was no animal performance benefit for replacing SFC with GLY in diets containing yellow grease, and the only change in carcass merit was a slight improvement in YG.

  11. A rapid and convenient method for specific 11C-labelling of synthetic polypeptides containing methionine

    Laengstroem, B.; Sjoeberg, S.; Ragnarsson, U.

    1981-01-01

    11 C-labelling of methionine residues in a synthetic peptide via the preparation of the corresponding protected, pure homocysteine peptide has been investigated. Complete deprotection of the peptide and specific methylation of the homocysteine residue can be performed in one step in liquid ammonia. As a first application of this method the synthesis of the tripeptide, Z-Gly-L-Hcy(Bzl)-Gly-O-Bzl, and its conversion to Gly-Met-Gly and the corresponding labelled Gly-([ 11 C]-methyl)-Met-Gly, is reported. Starting with the protected peptide the labelling was performed in 20 +- 5 min (starting with 11 CO 2 ), yielding the labelled peptide in 92 +- 5 % radiochemical yield. Analyses and preparative LC can be performed within 6 min. (author)

  12. Carbohydrate protease conjugates: Stabilized proteases for peptide synthesis

    Wartchow, C.A.; Wang, Peng; Bednarski, M.D.; Callstrom, M.R. [Ohio State Univ., Columbus, OH (United States)]|[Lawrence Berkeley Lab., CA (United States)

    1995-12-31

    The synthesis of oligopeptides using stable carbohydrate protease conjugates (CPCs) was examined in acetonitrile solvent systems. CPC[{alpha}-chymotrypsin] was used for the preparation of peptides containing histidine, phenylalanine, tryptophan in the P{sub 1} position in 60-93% yield. The CPC[{alpha}-chymotrypsin]-catalyzed synthesis of octamer Z-Gly-Gly-Phe-Gly-Gly-Phe-Gly-Gly-OEt from Z-Gly-Gly-Phe-Gly-Gly-Phe-OMe was achieved in 71% yield demonstrating that synthesis peptides containing both hydrophylic and hydrophobic amino acids. The P{sub 2} specificity of papain for aromatic residues was utilized for the 2 + 3 coupling of Z-Tyr-Gly-OMe to H{sub 2}N-Gly-Phe-Leu-OH to generate the leucine enkephalin derivative in 79% yield. Although papain is nonspecific for the hydrolysis of N-benzyloxycarbonyl amino acid methyl esters in aqueous solution, the rates of synthesis for these derivitives with nucleophile leucine tert-butyl ester differed by nearly 2 orders of magnitude. CPC[thermolysin] was used to prepare the aspartame precursor Z-Asp-Phe-OMe in 90% yield. The increased stability of CPCs prepared from periodate-modified poly(2-methacryl- amido-2-deoxy-D-glucose), poly(2-methacrylamido-2-deoxy-D-galactose), and poly(5-methacryl-amido-5-deoxy-D-ribose), carbohydrate materials designed to increase the aldehyde concentration in aqueous solution, suggests that the stability of CPCs is directly related to the aldehyde concentration of the carbohydrate material. Periodate oxidation of poly(2-methacrylamido-2-deoxy-D-glucose) followed by covalent attachment to {alpha}-chymotrypsin gave a CPC with catalytic activity in potassium phosphate buffer at 90{degrees}C for 2 h. 1 fig., 1 tab., 40 refs.

  13. Molecular sites for the positive allosteric modulation of glycine receptors by endocannabinoids.

    Gonzalo E Yévenes

    Full Text Available Glycine receptors (GlyRs are transmitter-gated anion channels of the Cys-loop superfamily which mediate synaptic inhibition at spinal and selected supraspinal sites. Although they serve pivotal functions in motor control and sensory processing, they have yet to be exploited as drug targets partly because of hitherto limited possibilities for allosteric control. Endocannabinoids (ECs have recently been characterized as direct allosteric GlyR modulators, but the underlying molecular sites have remained unknown. Here, we show that chemically neutral ECs (e.g. anandamide, AEA are positive modulators of α(1, α(2 and α(3 GlyRs, whereas acidic ECs (e.g. N-arachidonoyl-glycine; NA-Gly potentiate α(1 GlyRs but inhibit α(2 and α(3. This subunit-specificity allowed us to identify the underlying molecular sites through analysis of chimeric and mutant receptors. We found that alanine 52 in extracellular loop 2, glycine 254 in transmembrane (TM region 2 and intracellular lysine 385 determine the positive modulation of α(1 GlyRs by NA-Gly. Successive substitution of non-conserved extracellular and TM residues in α(2 converted NA-Gly-mediated inhibition into potentiation. Conversely, mutation of the conserved lysine within the intracellular loop between TM3 and TM4 attenuated NA-Gly-mediated potentiation of α(1 GlyRs, without affecting inhibition of α(2 and α(3. Notably, this mutation reduced modulation by AEA of all three GlyRs. These results define molecular sites for allosteric control of GlyRs by ECs and reveal an unrecognized function for the TM3-4 intracellular loop in the allosteric modulation of Cys-loop ion channels. The identification of these sites may help to understand the physiological role of this modulation and facilitate the development of novel therapeutic approaches to diseases such as spasticity, startle disease and possibly chronic pain.

  14. Frequency-dependent glycinergic inhibition modulates plasticity in hippocampus.

    Keck, Tara; Lillis, Kyle P; Zhou, Yu-Dong; White, John A

    2008-07-16

    Previous studies have demonstrated the presence of functional glycine receptors (GlyRs) in hippocampus. In this work, we examine the baseline activity and activity-dependent modulation of GlyRs in region CA1. We find that strychnine-sensitive GlyRs are open in the resting CA1 pyramidal cell, creating a state of tonic inhibition that "shunts" the magnitude of EPSPs evoked by electrical stimulation of the Schaffer collateral inputs. This GlyR-mediated shunting conductance is independent of the presynaptic stimulation rate; however, pairs of presynaptic and postsynaptic action potentials, repeated at frequencies above 5 Hz, reduce the GlyR-mediated conductance and increase peak EPSP magnitudes to levels at least 20% larger than those seen with presynaptic stimulation alone. We refer to this phenomenon as rate-dependent efficacy (RDE). Exogenous GlyR agonists (glycine, taurine) block RDE by preventing the closure of postsynaptic GlyRs. The GlyR antagonist strychnine blocks postsynaptic GlyRs under all conditions, occluding RDE. During RDE, GlyRs are less responsive to local glycine application, suggesting that a reduction in the number or sensitivity of membrane-inserted GlyRs underlies RDE. By extending the RDE induction protocol to include 500 paired presynaptic and postsynaptic spikes, we can induce long-term synaptic depression (LTD). Manipulations that lead to reduced functionality of GlyRs, either pharmacologically or through RDE, also lead to increased LTD. This result suggests that RDE contributes to long-term synaptic plasticity in the hippocampus.

  15. Positive/negative liquid secondary ion mass spectrometry of Ln-EDTA (1:1) complexes. Formation of molecular ion adducts with neutral species of the matrix or Ln-EDTA

    Plaziak, A.S.; Lis, S.; Elbanowski, M.

    1992-01-01

    The mass spectra of 1:1 complexes of EDTA with lanthanide cations (Ln=Sm, Eu, Gd, Tb or Dy) upon positive/negative LSIMS are presented. In glycerol used as a matrix, adduct-ions such as [M+H] + , [M+H+nGly] + , [2M+H] + , [2M+H+Gly] + (positive LSIMS) or [M-H] - , [M-H+nGly] - , [2M-H] - , [2M-H+Gly] - (negative LSIMS), where n=1-3, are formed. Reactions leading to the formation of adduct-ions are suggested. (authors)

  16. Generation of biogas using crude glycerin from biodiesel production as a supplement to cattle slurry

    Robra, S.; Neto, J. A. Almeida [Departamento de Ciencias Agrarias e Ambientais, Universidade Estadual de Santa Cruz, Rod. Ilheus/Itabuna km 16 s/n, CEP 45662-000 Ilheus, Bahia (Brazil); Serpa da Cruz, R.; de Oliveira, A.M.; Santos, J.V. [Departamento de Ciencias Exatas e Tecnologicas, Universidade Estadual de Santa Cruz, Rod. Ilheus/Itabuna km 16 s/n, CEP 45662-000 Ilheus, Bahia (Brazil)

    2010-09-15

    The influence of crude glycerin on biogas production and methane content of the produced biogas was studied, when added to cattle slurry. The experimental design consisted of 5% wt (Gli 5), 10% wt (Gli 10), and 15% wt (Gli 15) of crude glycerin added to cattle slurry, and one control digester without addition of crude glycerin. Anaerobic digestion was carried out in 4 laboratory size CSTR-type biogas digesters with a working volume of 3 L, in semi-continuous regime at mesophilic conditions, over a period of 10 weeks. The highest biogas yields (825.3 mL g{sup -1} and 825.7 mL g{sup -1}, respectively) relative to mass of volatile compounds added, were produced by the treatments Gli 5 and Gli 10. The control treatment produced 268.6 mL g{sup -1}, whereas the treatment Gli 15 produced 387.9 mL g{sup -1}. This low value was due to the breakdown of the process. Compared to the control, methane contents was increased by 9.5%, 14.3%, and 14.6%, respectively, for the treatments Gli 5, Gli 10, and Gli 15. (author)

  17. Towards an understanding of structure-function relationships of elongation factor Tu

    Wiborg, O; Andersen, C; Knudsen, Charlotte Rohde

    1994-01-01

    In light of the recently determined structure of elongation factor Tu, and taking into account chemical studies mapping functional sites, a number of residues have been selected for site-directed mutagenesis studies. Gly94, Gly126, His66, His118, Lys89 and Asp90 have each been point-mutated. Prel......In light of the recently determined structure of elongation factor Tu, and taking into account chemical studies mapping functional sites, a number of residues have been selected for site-directed mutagenesis studies. Gly94, Gly126, His66, His118, Lys89 and Asp90 have each been point...

  18. Enhancement of intestinal water absorption and sodium transport by glycerol in rats.

    Wapnir, R A; Sia, M C; Fisher, S E

    1996-12-01

    Glycerol (Gly) is a hydrophilic, absorbable, and energy-rich solute that could make water absorption more efficient. We investigated the use of Gly in a high-energy beverage containing corn syrup (CS) by using a small intestine perfusion procedure in the rat, an approach shown earlier to provide good preclinical information. The effectiveness of several formulations with Gly and CS was compared with commercial products and to experimental formulas where Gly substituted for glucose (Glc). The CS-Gly combination was more effective than preparations on the market containing sucrose and Glc-fructose syrups (G-P and G-L, respectively) in maintaining a net water absorption balance in the test jejunal segment [CS-Gly = 0.21 +/- 0.226, G-L = -1.516 +/- 0.467, and G-P = -0.299 +/- 0.106 (SE) microliter.min-1.cm-1 (P = 0.0113)] and in reducing sodium release into the lumen [CS-Gly = -133.2 +/- 16.2, G-L = -226.7 +/- 25.2, and G-P = -245.6 +/- 23.4 nmol.min-1.cm-1 (P = 0.0022)]. In other preparations, at equal CS concentrations (60 and 80 g/l, respectively), Gly clearly improved net water absorption over a comparable Glc-containing product [CS60-Gly = 0.422 +/- 0.136 and CS80-Gly = 0.666 +/- 0.378 vs. CS60-Glc = -0.282 +/- 0.200 and CS80-Glc = -1.046 +/- 0.480 microliters.min-1.cm-1 (P = 0.0019)]. On the basis of the data of this rat intestine perfusion model, Gly could be a useful ingredient in energy-rich beverages and might enhance fluid absorption in humans.

  19. Production and molecular characterization of bread wheat lines with reduced amount of α-type gliadins.

    Camerlengo, Francesco; Sestili, Francesco; Silvestri, Marco; Colaprico, Giuseppe; Margiotta, Benedetta; Ruggeri, Roberto; Lupi, Roberta; Masci, Stefania; Lafiandra, Domenico

    2017-12-19

    Among wheat gluten proteins, the α-type gliadins are the major responsible for celiac disease, an autoimmune disorder that affects about 1% of the world population. In fact, these proteins contain several toxic and immunogenic epitopes that trigger the onset of the disease. The α-type gliadins are a multigene family, encoded by genes located at the complex Gli-2 loci. Here, three bread wheat deletion lines (Gli-A2, Gli-D2 and Gli-A2/Gli-D2) at the Gli-2 loci were generated by the introgression in the bread wheat cultivar Pegaso of natural mutations, detected in different bread wheat cultivars. The molecular characterization of these lines allowed the isolation of 49 unique expressed genes coding α-type gliadins, that were assigned to each of the three Gli-2 loci. The number and the amount of α-type gliadin transcripts were drastically reduced in the deletion lines. In particular, the line Gli-A2/Gli-D2 contained only 12 active α-type gliadin genes (-75.6% respect to the cv. Pegaso) and a minor level of transcripts (-80% compared to cv. Pegaso). Compensatory pleiotropic effects were observed in the two other classes of gliadins (ω- and γ-gliadins) either at gene expression or protein levels. Although the comparative analysis of the deduced amino acid sequences highlighted the typical structural features of α-type gliadin proteins, substantial differences were displayed among the 49 proteins for the presence of toxic and immunogenic epitopes. The deletion line Gli-A2/Gli-D2 did not contain the 33-mer peptide, one of the major epitopes triggering the celiac disease, representing an interesting material to develop less "toxic" wheat varieties.

  20. Description of Three New α Variants and Four New β Variants: Hb Montluel [α110(G17)Ala → Val; HBA1: c.332C > T], Hb Cap d'Agde [α131(H14)Ser → Cys; HBA2: c.395C > G] and Hb Corsica [α100(G7)Leu → Pro; HBA1: 302T > C]; Hb Nîmes [β104(G6)Arg → Gly; HBB: c.313A > G], Hb Saint Marcellin [β112(G14)Cys → Gly; HBB: c.337T > G], Hb Saint Chamond [β80(EF4)Asn → 0; HBB: c.241_243delAAC] and Hb Dompierre [β29(B11)Gly → Arg; HBB: c.88G > C].

    Renoux, Céline; Feray, Cécile; Joly, Philippe; Lacan, Philippe; Francina, Alain

    2015-01-01

    We present here seven new hemoglobin (Hb) variants identified during routine Hb analysis. All of them are caused by a missense mutation except Hb Saint Chamond, which results from an in-frame deletion of the asparagine residue at β80. All these variants are clinically silent in the heterozygous state but two of them (Hb Cap d'Agde and Hb Dompierre) may be unstable, whereas Hb Nîmes could present a very slightly elevated oxygen affinity. These data are to be confirmed by appropriate biochemical tests.

  1. The lives and opinions of Socrates and Stilpo as defended by Plutarch against the insidious yet ignorant attacks of Colotes La vie et l'opinion de Socrate et de Stilpon défendues par Plutarque contre les attaques insidieuses mais ignorantes de Colotès Le vite e le opinioni di Socrate e Stilpo come difesa da Plutarco contro gli attacchi insidiosi ancora ignoranti di Colote

    Jan Opsomer

    2013-05-01

    Full Text Available Dans les chapitres 17 à 23 de l’Adversus Colotem, Plutarque défend Socrate et Stilpon contre les critiques de Colotès. J’examine l’insertion de cette section dans le contexte plus large de l’œuvre, les liens entre la section sur Socrate et Stilpon, les critiques de Colotès et les contre-arguments ainsi que les stratégies polémiques de Plutarque. Ce faisant, je tente de démêler les différentes couches de ce texte complexe. Socrate est présenté comme un imposteur et un sceptique par Colotès. Plutarque affirme que la philosophie de Socrate implique certes une méfiance radicale à l’égard des sens, mais que cela n’empêche ni Socrate ni ses disciples de vivre leur vie quotidienne. La philosophie socratique doit être préférée de très loin à l’épicurisme. Stilpon, un mégarique, appartient également à la tradition socratique. Plutarque apporte son soutien à la bonne réputation morale de ce philosophe à l’esprit vif contre le mauvais traitement que Colotès lui avait infligé. Ce dernier avait aussi attaqué le rejet par Stilpon de toute prédication, à l’exception de la prédication d’identité. Plutarque affirme que l’argument de Stilpon est un pur exercice dialectique et ne menace aucunement nos vies quotidiennes. Plutarque lance en outre une contre-attaque contre la philosophie épicurienne du langage, et plus particulièrement la suppression du niveau intermédiaire – celui des significations. Le fameux argument de Stilpon, tel qu’il est présenté par Plutarque, revient à nier la relation ontologique à laquelle correspond la prédication ordinaire. La conclusion de Plutarque – selon laquelle Stilpon nous invite simplement à abandonner l’usage du verbe « être » comme copule – ne convient pas à l’argument tel qu’il le présente. La meilleure explication de ce décalage est l’hypothèse que Plutarque a copié l’argument de manière assez fidèle sans l’analyser dans le détail. Cela expliquerait également le fait que l’argument ne fasse aucun usage de la théorie platonicienne de la prédication dans le Sophiste.Nei capitoli 17-23 del Contro Colote Plutarco difende Socrate e Stilpone dalla critica di Colote. Esamino l’inserimento di questa sezione nel più ampio contesto dell’opera, i legami fra le sezioni dedicate a Socrate e a Stilpone, le critiche di Colote e i controargomenti e le strategie polemiche di Plutarco. Nel fare ciò, mi propongo di distinguere i differenti strati che compongono questo complesso testo. Socrate è rappresentato da Colote come un impostore e uno scettico. Plutarco sostiene che la filosofia socratica implica, sì, una sfiducia radicale nei confronti dei sensi, ma questo non rende impossibile a lui e ai suoi seguaci di vivere la loro vita quotidiana. La filosofia socratica va preferita di gran lunga all’epicureismo. Anche Stilpone, un megarese, appartiene alla tradizione socratica. Plutarco difende l’alta raputazione morale di questo acuto filosofo dagli insulti ricevuti da Colote. Quest’ultimo aveva attaccato anche il rigetto, da parte di Stilpone, di ogni predicazione che non fosse la predicazione di identità. Plutarco afferma che l’argomento di Stiplone è un mero esercizio dialettico che non pone alcuna minaccia alle nostre vite quotidiane. Inoltre, Plutarco si cimenta in un contrattacco alla filosofia epicurea del linguaggio, e in particolare la sua abolizione del livello intermedio – quello dei significati. Quando Plutarco presenta il celebre argomento di Stilpone, esso risulta in una negazione della relazione ontologica a cui corrisponde la predicazione ordinaria. La conclusione plutarchea – Stilpone ci invita soltanto ad abbandonare l’uso del verbo ‘essere’ come copula – non collima con l’argomento così come esso è presentato. Questo scarto può essere spiegato nel modo più plausibile ipotizzando che Plutarco abbia copiato l’argomento piuttosto fedelmente senza analizzarlo in dettaglio. Questo spiegherebbe anche il fatto che l’argomento non faccia alcun uso del resoconto che Platone fa della predicazione nel Sofista.In chapters 17 to 23 of the Adversus Colotem Plutarch defends Socrates and Stilpo against Colotes’ criticism. I examine the embedding of this section in the larger context of the work, the links between the sections on Socrates and Stilpo, Colotes’ criticisms and Plutarch’s counter-arguments and polemical strategies. In doing so, I attempt to disentangle the different layers of this complex text. Socrates is portrayed as an imposter and a sceptic by Colotes. Plutarch argues that Socrates’ philosophy indeed implies a radical distrust of the senses, but that this does not make it impossible for him and his followers to live their daily lives. Socratic philosophy is much to be preferred to Epicureanism. Stilpo, a Megarian, also belongs to the Socratic tradition. Plutarch upholds the high moral reputation of this sharp-witted philosopher against the abuse that he had to suffer from Colotes. The latter also attacked Stilpo’s rejection of all predication except identity predication. Plutarch claims that Stilpo’s argument is a mere dialectical exercise and poses no threat to our daily lives. Plutarch moreover engages in a counter-attack on the Epicurean philosophy of language, more particularly their abolishment of the intermediate level – that of meanings. As Plutarch presents Stilpo’s famous argument, it amounts to a denial of the ontological relation to which ordinary predication corresponds. Plutarch’s conclusion – that Stilpo merely urges us to abandon the use of the verb “to be” as a copula – does not fit the argument as it is presented by him. This mismatch is best explained by the hypothesis that Plutarch copied the argument quite faithfully without analysing it in detail. That would also explain the fact that the argument makes no use of Plato’s account of predication in the Sophist.

  2. Tritium labelling of two highly selective agonists for CCK-B receptors : [[sup 3]H]propionyl-Tyr(SO[sub 3]Na)-gNle-mGly-Trp-(N-Me)Nle-Asp-Phe-NHsub (2) ([[sup 3]H]pBC 264) [[sup 3]H]propionyl-[gamma]D. Glu-Tyr(SO[sub 3]H)-Nle-D. Lys-Trp-Nle-Asp-Phe-NH[sub 2] ([[sup 3]H]pBC 254). [Cholecystokini-B receptors

    Corringer, P J; Durieux, C; Ruiz-Gayo, M; Roques, B P [UA498 CNRS, U266 INSERM, UFR des Sciences Pharmaceutiques et Biologiques, 75 - Paris (France)

    1992-06-01

    Among the CCK-B receptor agonists reported to date, the two modified peptides BC 264 and BC 254 display a high affinity and selectivity for this binding site and are highly protected from enzymatic degradation. Recently, we reported the biological properties of a tritiated analog of this agonist, [[sup 3]H]pBC 264, which fullfils all the criteria required for in vitro as well as in vivo studies of the CCK-B receptor. On the other hand, BC 254 displays a high affinity for the CCK-B binding sites in the guinea-pig (K[sub i] = 0.56 nM) while its affinity in the rat is more than 60-fold lower, a difference which could be due to the occurrence of CCK-B receptor subtypes. In the present paper, we report the synthesis of [[sup 3]H]pBC 264 and of the new tritiated ligand [[sup 3]H]pBC 254 using [[sup 3]H] NPS (N-succinimidyl[2,3-[sup 3]H]propionate) as labelling agent. These two probes have high specific activity (70-100 Ci/mmol) and will enable extensive studies of the CCK-B receptors to be carried out. (author).

  3. Crystallization and preliminary X-ray diffraction analysis of an Escherichia coli tRNAGly acceptor-stem microhelix

    Förster, Charlotte; Perbandt, Markus; Brauer, Arnd B. E.; Brode, Svenja; Fürste, Jens P.; Betzel, Christian; Erdmann, Volker A.

    2006-01-01

    In order to investigate the identity elements of the E. coli tRNA Gly /GlyRS class II system, a tRNA Gly acceptor-stem microhelix was crystallized and a data set was collected to 2.0 Å resolution using synchrotron radiation. The tRNA Gly and glycyl-tRNA synthetase (GlyRS) system is an evolutionary special case within the class II aminoacyl-tRNA synthetases because two divergent types of GlyRS exist: an archaebacterial/human type and an eubacterial type. The tRNA identity elements which determine the correct aminoacylation process are located in the aminoacyl domain of tRNA Gly . To obtain further insight concerning structural investigation of the identity elements, the Escherichia coli seven-base-pair tRNA Gly acceptor-stem helix was crystallized. Data were collected to 2.0 Å resolution using synchrotron radiation. Crystals belong to space group P3 1 21 or P3 2 21, with unit-cell parameters a = b = 35.35, c = 130.82 Å, α = β = 90, γ = 120° and two molecules in the asymmetric unit

  4. Lo Strategic Management Accounting

    G. INVERNIZZI

    2005-01-01

    Il saggio indaga gli aggregati informativi e gli elementi che compongono lo strategic management accounting. Sono quindi analizzate le funzioni svolte nei diversi stadi del processo di gestione strategica osservando il suo ruolo all’interno del management accounting. Infine sono approfonditi i rapporti fra i livelli della gestione strategica e lo strategic management accounting.

  5. Spop promotes skeletal development and homeostasis by positively regulating Ihh signaling.

    Cai, Hongchen; Liu, Aimin

    2016-12-20

    Indian Hedgehog (Ihh) regulates chondrocyte and osteoblast differentiation through the Glioma-associated oncogene homolog (Gli) transcription factors. Previous in vitro studies suggested that Speckle-type POZ protein (Spop), part of the Cullin-3 (Cul3) ubiquitin ligase complex, targets Gli2 and Gli3 for degradation and negatively regulates Hedgehog (Hh) signaling. In this study, we found defects in chondrocyte and osteoblast differentiation in Spop-null mutant mice. Strikingly, both the full-length and repressor forms of Gli3, but not Gli2, were up-regulated in Spop mutants, and Ihh target genes Patched 1 (Ptch1) and parathyroid hormone-like peptide (Pthlh) were down-regulated, indicating compromised Hh signaling. Consistent with this finding, reducing Gli3 dosage greatly rescued the Spop mutant skeletal defects. We further show that Spop directly targets the Gli3 repressor for ubiquitination and degradation. Finally, we demonstrate in a conditional mutant that loss of Spop results in brachydactyly and osteopenia, which can be rescued by reducing the dosage of Gli3. In summary, Spop is an important positive regulator of Ihh signaling and skeletal development.

  6. The intracellular domain of the Drosophila cholinesterase-like neural adhesion protein, gliotactin, is natively unfolded

    Zeev-Ben-Mordehai, T; Rydberg, EH; Solomon, A.; Toker, L; Auld, VJ; Silman, I.; Botti, S; Sussman, J.L.

    2003-01-01

    Drosophila gliotactin (Gli) is a 109-kDa transmembrane, cholinesterase-like adhesion molecule (CLAM), expressed in peripheral glia, that is crucial for formation of the blood-nerve barrier. The intracellular portion (Gli-cyt) was cloned and expressed in the cytosolic fraction of Escherichia coli

  7. Thermodynamics of formation for the 18-crown-6-triglycine molecular complex in water-dimethylsulfoxide solvents

    Usacheva, T. R.; Lan, Pham Thi; Sharnin, V. A.

    2014-06-01

    The effect of a water-dimethylsulfoxide (DMSO) solvent on the formation of a molecular complex of 18-crown-6 (18C6) with triglycine (diglycylglycine, 3Gly) is studied via calorimetric titration. It is found that switching from water to an H2O-DMSO mixture with DMSO mole fraction of 0.30 is accompanied by a monotonic increase in the stability of [3Gly18C6] complex, from log K ∘ = 1.10 to log K ∘ = 2.44, and an increase in the exothermicity of the reaction of its formation, from -5.9 to -16.9 kJ/mol. It is shown that the [3Gly18C6] complex exhibits enthalpy stabilization with negative values of enthalpy and entropy over the investigated range of H2O-DMSO solvents. Analysis of the reagents' solvation characteristics reveals that the increase in the reaction's exothermicity of transfer is due to differences in the solvation of [3Gly18C6] and 18C6 with a small solvation contribution from 3Gly. It is concluded that the change in the Gibbs energy of the reaction 3Glysolv + 18C6solv ↔ [3Gly18C6]solv is due to differences in the change in the solvation state of the complex and the peptide (Δtr G ∘([3Gly18C6])-Δtr G ∘(3Gly)).

  8. AcEST: DK947950 [AcEST

    Full Text Available 10071|GLI3_HUMAN Zinc finger protein GLI3 OS=Homo sapiens GN... 31 4.9 sp|Q8WXX7|AUTS2_HUMAN Autism suscepti...FSPPHPYINPYMDYIRSLHSSPSLSMI 216 >sp|Q8WXX7|AUTS2_HUMAN Autism susceptibility gene 2 protein OS=Homo sapiens

  9. Horseradish peroxidase-catalyzed oligomerization of ferulic acid on a template of a tyrosine-containing tripeptide

    Oudgenoeg, G.; Dirksen, E.; Ingemann, S.; Hilhorst, R.; Gruppen, H.; Boeriu, C.G.; Piersma, S.R.; Berkel, W.J.H. van; Laane, C.; Voragen, A.G.J.

    2002-01-01

    Ferulic acid (FA) is an abundantly present phenolic constituent of plant cell walls. Kinetically controlled incubation of FA and the tripeptide Gly-Tyr-Gly (GYG) with horseradish peroxidase and H2O2 yielded a range of new cross-linked products. Two predominant series of hetero-oligomers of FA linked

  10. Drug: D09937 [KEGG MEDICUS

    Full Text Available cal group: DG01344 ... syntheses Corticotropin (ACTH) [CPD:C02017] MC1R [HSA:4157] [KO:K04199]; MC2R [HSA:4158...e Arg Trp Gly Lys Pro Val Gly Lys Lys Arg Arg-NH2 ... Other ... DG01629 ... Adrenocorticotropic hormone (ACTH) Chemi

  11. Disturbances of water, electrolyte and acid-base metabolism in ...

    Chantel

    tion of intravenous sodium bicarbonate, initially at doses of no more than 1 - 2 mmol/kg of body weight (see information below). • Acute poisonings. These include methanol, ethylene gly- col, and salicylate poisoning. Both methanol and ethylene gly- col are substrates for hepatic alcohol dehydrogenase and are metabolised ...

  12. Gluconeogenesis during endurance exercise in cyclists habituated to a long-term low carbohydrate high fat diet

    Endogenous glucose production (EGP) occurs via hepatic glycogenolysis (GLY) and gluconeogenesis (GNG) and plays an important role in maintaining euglycemia. Rates of GLY and GNG increase during exercise in athletes following a mixed macronutrient diet; however these processes have not been investiga...

  13. Lysosomally cleavable peptide-containing polymersomes modified with anti-EGFR antibody for systemic cancer chemotherapy

    Lee, Jung Seok; Groothuis, T.A.M.; Cusan, Claudia; Mink, Daniel; Feijen, Jan

    2011-01-01

    Polymersomes (Ps) based on a biodegradable and biocompatible block copolymer of methoxy poly(ethylene glycol) (mPEG) and poly(d,l-lactide) (PDLLA) in which apeptide sequence, Gly-Phe-Leu-Gly-Phe (GFLGF), was introduced in between the two blocks(mPEG-pep-PDLLA) were developed. The peptide linker is

  14. Development of an Electrochemical-Cantilever Hybrid Platform

    Fischer, Lee MacKenzie

    . For at binde kobber (II) ioner blev rækker af cantilevere funktionaliseret med aminosyre L-cysteine(Cys) og tetrapeptid Cys-Gly-Gly-His (CGGH). Dette funktionelle lag blev fjernet fra en enkelt cantilever, ved selektivt at anvende et voltammetrisk signal til at generere en ren reference cantilever til brug...

  15. Genotyping for NOD2 genetic variants and crohn disease: a metaanalysis

    Yazdanyar, Shiva; Weischer, Maren; Nordestgaard, Børge

    2009-01-01

    BACKGROUND: Arg702Trp, Gly908Arg, and Leu1007fsinsC variants of the NOD2 gene (nucleotide-binding oligomerization domain containing 2; alias, CARD15) influence the risk of Crohn disease. METHODS: We conducted a systematic review to examine whether Arg702Trp, Gly908Arg, and Leu1007fsinsC are equally...

  16. Dicty_cDB: Contig-U15594-1 [Dicty_cDB

    Full Text Available e, intron; ... 32 1.7 4 ( AY163064 ) Bryum capillare tRNA-Gly (trnG) gene, intron sequ... 32 1.7 4 ( EF362523 ) Plagio...bryum capillare tRNA-Gly (trnG) gene, intro... 32 1.7 4 ( DQ536453 ) Plagiobryum capillare tRNA-Gl

  17. luce profuga di valerio aiolli e la badante. un amore involontario di ...

    User

    with Italians are superficial. The novels Luce ... in queste opere appaiono gli stessi stereotipi che si trovano nei media, cioè gli stranieri .... Georg Simmel, “The Stranger” in On Individual and Social Forms, University of Chicago Press: Chicago ...

  18. Oxytocin biotransformation in the rat limbic brain

    Burbach, J.P.H.; Schotman, P.; Kloet, E.R. de

    2006-01-01

    Two peptide fragments of oxytocin were isolated by high-pressure liquid chromatography from digests of oxytocin obtained after exposure to a SPM preparation of the rat limbic brain. The structures of these peptides, being Gln-Asn-Cys(O)x-Pro-Leu-GlyNH2 and Gln-Asn-Cys(-S-S-Cys)-Pro-Leu-GlyNH2, were

  19. Biografia della fisica

    Gamow, George

    1963-01-01

    Nato a Odessa nel 1904, è morto a Boulder (Colorado) nel 1968. Dopo essersi laureato all'Università di Leningrado, nel 1928 frequentò la scuola estiva di Gottinga. Le ricerche di quel periodo gli consentirono di spegiare il fenomeno, allora misterioso, della radioattività naturale e gli esperimenti di Lord Rutherford sulla trasformazione indotta sugli elementi leggeri...

  20. Passive versus active tumor targeting using RGD- and NGR-modified polymeric nanomedicines

    Kunjachan, Sijumon; Pola, Robert; Gremse, Felix; Theek, Benjamin; Ehling, Josef; Moeckel, Diana; Hermanns-Sachweh, Benita; Pechar, Michal; Ulbrich, Karel; Hennink, Wim E.; Storm, Gert; Lederle, Wiltrud; Kiessling, Fabian; Lammers, Twan

    2014-01-01

    Enhanced permeability and retention (EPR) and the (over-) expression of angiogenesis-related surface receptors are key features of tumor blood vessels. As a consequence, EPR-mediated passive and Arg-Gly-Asp (RGD) and Asn-Gly-Arg (NGR) based active tumor targeting have received considerable attention

  1. Serre fotovoltaiche Attenti alle perdite; Stanghellini: il calo produttivo non è da poco

    Contri, L.; Stanghellini, C.

    2010-01-01

    Gli installatori di pannelli fotovoltaici hanno nel mirino gli ortoflorovivaisti che gestiscono serre. Costoro, per contro, sono alla ricerca di sistemi atti a far loro recuperare la marginalità perduta in questi anni di crisi. Un matrimonio che s'ha da fare? Meglio di no. A sconsigliarlo è Cecilia

  2. Cucurbit[8]uril templated supramolecular ring structure formation and protein assembly modulation

    Ramaekers, M.; Wijnands, S.P.W.; van Dongen, J.L.J.; Brunsveld, L.; Dankers, P.Y.W.

    2015-01-01

    The interplay of Phe-Gly-Gly (FGG)-tagged proteins and bivalent FGG-tagged penta(ethylene glycol) as guest molecules with cucurbit[8]uril (Q8) hosts is studied to modulate the supramolecular assembly process. Ring structure formation of the bivalent guest molecule with Q8 leads to enhanced binding

  3. The Battle of Vukovar: The Battle That Saved Croatia

    2002-07-01

    1991, 4. 53 M. Marjanovic and D. Glišic, “Visoka cena slobode” [The High Price of Freedom], Narodna armija (Belgrade), 21 September 1991, 10. 39...The Bull Has No More Hay”] Ilustrovana politika, (Belgrade) 9 October 1991. o) Marjanovic M. and Glišic D., “Visoka cena slobode” [The High Price of

  4. Trypanosoma brucei TBRGG1, a mitochondrial oligo(U)-binding protein that co-localizes with an in vitro RNA editing activity

    Vanhamme, L.; Perez-Morga, D.; Marchal, C.; Speijer, D.; Lambert, L.; Geuskens, M.; Alexandre, S.; Ismaïli, N.; Göringer, U.; Benne, R.; Pays, E.

    1998-01-01

    We report the characterization of a Trypanosoma brucei 75-kDa protein of the RGG (Arg-Gly-Gly) type, termed TBRGG1. Dicistronic and monocistronic transcripts of the TBRGG1 gene were produced by both alternative splicing and polyadenylation. TBRGG1 was found in two or three forms that differ in their

  5. STANDARDIZATION OF GLYCOHEMOGLOBIN RESULTS AND REFERENCE VALUES IN WHOLE-BLOOD STUDIED IN 103 LABORATORIES USING 20 METHODS

    WEYKAMP, CW; PENDERS, TJ; MUSKIET, FAJ; VANDERSLIK, W

    We investigated the effect of calibration with lyophilized calibrators on whole-blood glycohemoglobin (glyHb) results. One hundred three laboratories, using 20 different methods, determined glyHb in two lyophilized calibrators and two whole-blood samples. For whole-blood samples with low (5%) and

  6. Prediction of solvent dependent Beta-roll formation of a self-assembling silk-like protein domain

    Schor, M.; Martens, A.A.; Wolf, de F.A.; Cohen Stuart, M.A.; Bolhuis, P.G.

    2009-01-01

    Triblock copolymers consisting of a middle silk-like [(Gly-Ala)3-Gly-Glu]n block flanked by two hydrophilic end blocks that assume a random conformation in aqueous solution at all pH spontaneously assemble into micrometre long fibers at low pH. As elucidating the molecular structure of the stacked

  7. Prediction of solvent dependent beta-roll formation of a self-assembling silk-like protein domain

    Schor, M.; Martens, A.A.; de Wolf, F.A.; Cohen Stuart, M.A.; Bolhuis, P.G.

    2009-01-01

    Triblock copolymers consisting of a middle silk-like [(Gly - Ala)(3) - Gly - Glu](n) block flanked by two hydrophilic end blocks that assume a random conformation in aqueous solution at all pH spontaneously assemble into micrometre long fibers at low pH. As elucidating the molecular structure of the

  8. Journal of Biosciences | Indian Academy of Sciences

    Home; Journals; Journal of Biosciences. Sangappa. Articles written in Journal of Biosciences. Volume 30 Issue 2 March 2005 pp 259-268 Articles. Crystal structure of raw pure Mysore silk fibre based on (Ala-Gly)2-Ser-Gly peptide sequence using Linked-Atom-Least-Squares method · Sangappa S S Mahesh R Somashekar.

  9. Polymorphism Glu389Arg of β1-adrenoreceptor gene and cardiovascular complications of hyperthyroidism

    A Yu Babenko

    2010-06-01

    Full Text Available Gly389Arg polymorphism β1-adrenoreceptors can influence the cardio-vascular prognosis. Human heart β1-adrenoreceptors perform a crucial role in mediating the cardiostimulant effects of norepinephrine. Understanding the significance of Gly389Arg polymorphism in the human heart is beginning to emerge, but not in adult patients with thyrotoxicosis. We've studied the Gly389Arg polymorphism of β1-adrenoreceptors gene in relation to Echo-cardiography parameters in 136 normotensive patients with a thyrotoxicosis without any CVD. Echo-CG was performed according to standard protocol before and during the thyreostatic treatment. The genotype distribution was as following: Gly/Gly – 25% (1 group (1 gr., Arg/Gly – 75% (2 group (2 gr., Arg/Arg – 0%. There was significant difference between 1 and 2 gr. by relative left ventricle wall thickness, left ventricular mass index, isovolumic relaxation time, Е/А ratio. The frequency of diastolic dysfunction (DD was in gr. 1–10%, in gr. 2–30%, р <0.001. After treatment during a year this damages were saved. These data demonstrate, that Gly/Gly genotype of β1-adrenoreceptors gene can have cardioprotective effect leading to less of LV hypertrophy and diastolic dysfunction in patients with thyrotoxicosis.

  10. Size-Dependent Affinity of Glycine and Its Short Oligomers to Pyrite Surface: A Model for Prebiotic Accumulation of Amino Acid Oligomers on a Mineral Surface

    Afrin, Rehana; Ganbaatar, Narangerel; Aono, Masashi; Cleaves, H. James; Yano, Taka-aki; Hara, Masahiko

    2018-01-01

    The interaction strength of progressively longer oligomers of glycine, (Gly), di-Gly, tri-Gly, and penta-Gly, with a natural pyrite surface was directly measured using the force mode of an atomic force microscope (AFM). In recent years, selective activation of abiotically formed amino acids on mineral surfaces, especially that of pyrite, has been proposed as an important step in many origins of life scenarios. To investigate such notions, we used AFM-based force measurements to probe possible non-covalent interactions between pyrite and amino acids, starting from the simplest amino acid, Gly. Although Gly itself interacted with the pyrite surface only weakly, progressively larger unbinding forces and binding frequencies were obtained using oligomers from di-Gly to penta-Gly. In addition to an expected increase of the configurational entropy and size-dependent van der Waals force, the increasing number of polar peptide bonds, among others, may be responsible for this observation. The effect of chain length was also investigated by performing similar experiments using l-lysine vs. poly-l-lysine (PLL), and l-glutamic acid vs. poly-l-glutamic acid. The results suggest that longer oligomers/polymers of amino acids can be preferentially adsorbed on pyrite surfaces. PMID:29370126

  11. Degradation of peptides by gamma-irradiation, 2

    Oku, Tadatake; Yoshida, Shigeki; Kondo, Mitsumasa; Ishida, Tomoharu; Fukui, Manabu; Ito, Teiichiro (Nihon Univ., Tokyo (Japan). Coll. of Agriculture and Veterinary Medicine)

    1990-10-01

    The radiolytic products of two kinds of dipeptides containing aromatic amino acid, gly-L-tyr and L-tyr-gly in 1 mM aqueous solution in the presence of air were examined by gamma-irradiation at doses of about 6, 12 and 25 kGy. Peptide samples in aqueous solution were analyzed by HPLC and GC after gamma-irradiation. Amides which the amounts of formation was very small, were collected several times by an amino acid autoanalyzer and isolated by HPLC. The ninhydrin-positive products from gly-L-tyr were detected gly, tyr, dopa, asp, ammonia, methylamine, ethylamine and glycinamide. The products from L-tyr-gly were tyr, gly, dopa, asp, ammonia, methylamine and ethylamine, but tyrosinamide was not confirmed. The total amounts of ninhydrin-positive products formed were less than the decreasing amount of each peptide at every irradiation dose. Methanal and ethanal were detected in both peptides. A radiolytic pathway of gly-L-tyr and L-tyr-gly was estimated from these results. (author).

  12. Molecular determinants of ivermectin sensitivity at the glycine receptor chloride channel

    Lynagh, Timothy; Webb, Timothy I.; Dixon, Christine L.

    2011-01-01

    Ivermectin is an anthelmintic drug that works by activating glutamate-gated chloride channel receptors (GluClRs) in nematode parasites. GluClRs belong to the Cys-loop receptor family that also includes glycine receptor (GlyR) chloride channels. GluClRs and A288G mutant GlyRs are both activated...

  13. Defining the phenotypic spectrum of SLC6A1 mutations

    Johannesen, Katrine M.; Gardella, Elena; Linnankivi, Tarja

    2018-01-01

    /polyspikes-and-slow waves in 25/31. Two patients developed an EEG pattern resembling electrical status epilepticus during sleep. Ataxia was observed in 7/34 cases. We describe 7 truncating and 18 missense variants, including 4 recurrent variants (Gly232Val, Ala288Val, Val342Met, and Gly362Arg). Significance: Most patients...

  14. A Novel Role for Keratin 17 in Coordinating Oncogenic Transformation and Cellular Adhesion in Ewing Sarcoma

    Sankar, Savita; Tanner, Jason M.; Bell, Russell; Chaturvedi, Aashi; Randall, R. Lor; Beckerle, Mary C.

    2013-01-01

    Oncogenic transformation in Ewing sarcoma is caused by EWS/FLI, an aberrant transcription factor fusion oncogene. Glioma-associated oncogene homolog 1 (GLI1) is a critical target gene activated by EWS/FLI, but the mechanism by which GLI1 contributes to the transformed phenotype of Ewing sarcoma was unknown. In this work, we identify keratin 17 (KRT17) as a direct downstream target gene upregulated by GLI1. We demonstrate that KRT17 regulates cellular adhesion by activating AKT/PKB (protein kinase B) signaling. In addition, KRT17 is necessary for oncogenic transformation in Ewing sarcoma and accounts for much of the GLI1-mediated transformation function but via a mechanism independent of AKT signaling. Taken together, our data reveal previously unknown molecular functions for a cytoplasmic intermediate filament protein, KRT17, in coordinating EWS/FLI- and GLI1-mediated oncogenic transformation and cellular adhesion in Ewing sarcoma. PMID:24043308

  15. Characterizing the Secondary Protein Structure of Black Widow Dragline Silk Using Solid-State NMR & X-ray Diffraction

    Jenkins, Janelle E.; Sampath, Sujatha; Butler, Emily; Kim, Jihyun; Henning, Robert W.; Holland, Gregory P.; Yarger, Jeffery L.

    2013-01-01

    This study provides a detailed secondary structural characterization of major ampullate dragline silk from Latrodectus hesperus (black widow) spiders. X-ray diffraction results show that the structure of black widow major ampullate silk fibers is comprised of stacked β-sheet nanocrystallites oriented parallel to the fiber axis and an amorphous region with oriented (anisotropic) and isotropic components. The combination of two-dimensional (2D) 13C-13C through-space and through-bond solid-state NMR experiments provide chemical shifts that are used to determine detailed information about amino acid motif secondary structure in black widow spider dragline silk. Individual amino acids are incorporated into different repetitive motifs that make up the majority of this protein-based biopolymer. From the solid-state NMR measurements, we assign distinct secondary conformations to each repetitive amino acid motif and hence to the amino acids that make up the motifs. Specifically, alanine is incorporated in β-sheet (poly(Alan) and poly(Gly-Ala)), 31-helix (poly(Gly-Gly-Xaa), and α-helix (poly(Gln-Gln-Ala-Tyr)) components. Glycine is determined to be in β-sheet (poly(Gly-Ala)) and 31-helical (poly(Gly-Gly-Xaa)) regions, while serine is present in β-sheet (poly(Gly-Ala-Ser)), 31-helix (poly(Gly-Gly-Ser)), and β-turn (poly(Gly-Pro-Ser)) structures. These various motif-specific secondary structural elements are quantitatively correlated to the primary amino acid sequence of major ampullate spidroin 1 and 2 (MaSp1 and MaSp2) and are shown to form a self-consistent model for black widow dragline silk. PMID:24024617

  16. Mass Spectrometry Identification of N-Chlorinated Dipeptides in Drinking Water.

    Huang, Guang; Jiang, Ping; Li, Xing-Fang

    2017-04-04

    We report the identification of N-chlorinated dipeptides as chlorination products in drinking water using complementary high-resolution quadrupole time-of-flight (QTOF) and quadrupole ion-trap mass spectrometry techniques. First, three model dipeptides, tyrosylglycine (Tyr-Gly), tyrosylalanine (Tyr-Ala), and phenylalanylglycine (Phe-Gly), reacted with sodium hypochlorite, and these reaction solutions were analyzed by QTOF. N-Cl-Tyr-Gly, N,N-di-Cl-Tyr-Gly, N-Cl-Phe-Gly, N,N-di-Cl-Phe-Gly, N-Cl-Tyr-Ala, and N,N-di-Cl-Tyr-Ala were identified as the major products based on accurate masses, 35 Cl/ 37 Cl isotopic patterns, and MS/MS spectra. These identified N-chlorinated dipeptides were synthesized and found to be stable in water over 10 days except N,N-di-Cl-Phe-Gly. To enable sensitive detection of N-chlorinated dipeptides in authentic water, we developed a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method with multiple reaction monitoring (MRM) mode. N-Cl-Tyr-Gly, N,N-di-Cl-Tyr-Gly, N-Cl-Phe-Gly, N-Cl-Tyr-Ala, and N,N-di-Cl-Tyr-Ala along with their corresponding dipeptides were detected in authentic tap water samples. The dipeptides were clearly detected in the raw water, but the N-chlorinated dipeptides were at background levels. These results suggest that the N-chlorinated dipeptides are produced by chlorination. This study has identified N-chlorinated dipeptides as new disinfection byproducts in drinking water. The strategy developed in this study can be used to identify chlorination products of other peptides in drinking water.

  17. Hedgehog signaling pathway in neuroblastoma differentiation.

    Souzaki, Ryota; Tajiri, Tatsuro; Souzaki, Masae; Kinoshita, Yoshiaki; Tanaka, Sakura; Kohashi, Kenichi; Oda, Yoshinao; Katano, Mitsuo; Taguchi, Tomoaki

    2010-12-01

    The hedgehog (Hh) signaling pathway is activated in some adult cancers. On the other hand, the Hh signaling pathway plays an important role in the development of the neural crest in embryos. The aim of this study is to show the activation of Hh signaling pathway in neuroblastoma (NB), a pediatric malignancy arising from neural crest cells, and to reveal the meaning of the Hh signaling pathway in NB development. This study analyzed the expression of Sonic hedgehog (Shh), GLI1, and Patched 1 (Ptch1), transactivators of Hh signaling pathway, by immunohistochemistry in 82 NB and 10 ganglioneuroblastoma cases. All 92 cases were evaluated for the status of MYCN amplification. Of the 92 cases, 67 (73%) were positive for Shh, 62 cases (67%) were positive for GLI1, and 73 cases (79%) were positive for Ptch1. Only 2 (10%) of the 20 cases with MYCN amplification were positive for Shh and GLI1, and 4 cases (20%) were positive for Ptch1 (MYCN amplification vs no MYCN amplification, P ≦ .01). The percentage of GLI1-positive cells in the cases with INSS stage 1 without MYCN amplification was significantly higher than that with INSS stage 4. Of 72 cases without MYCN amplification, 60 were GLI1-positive. Twelve cases were GLI1-negative, and the prognosis of the GLI1-positive cases was significantly better than that of the GLI1-negative cases (P = .015). Most of NBs without MYCN amplification were positive for Shh, GLI1, and Ptch1. In the cases without MYCN amplification, the high expression of GLI1 was significantly associated with early clinical stage and a good prognosis of the patients. In contrast to adult cancers, the activation of the Hh signaling pathway in NB may be associated with the differentiation of the NB. Copyright © 2010 Elsevier Inc. All rights reserved.

  18. In vivo evaluation of carbon-11-labelled non-sarcosine-based glycine transporter 1 inhibitors in mice and conscious monkeys

    Toyohara, Jun [Division of Clinical Neuroscience, Chiba University Center for Forensic Mental Health, Chiba, Japan 260-8670 (Japan); Positron Medical Center, Tokyo Metropolitan Institute of Gerontology, Tokyo, Japan 173-0022 (Japan); Ishiwata, Kiichi; Sakata, Muneyuki [Positron Medical Center, Tokyo Metropolitan Institute of Gerontology, Tokyo, Japan 173-0022 (Japan); Wu, Jin [Division of Clinical Neuroscience, Chiba University Center for Forensic Mental Health, Chiba, Japan 260-8670 (Japan); Nishiyama, Shingo; Tsukada, Hideo [Central Research Laboratory, Hamamatsu Photonics K.K., Shizuoka, Japan 434-8601 (Japan); Hashimoto, Kenji, E-mail: hashimoto@faculty.chiba-u.j [Division of Clinical Neuroscience, Chiba University Center for Forensic Mental Health, Chiba, Japan 260-8670 (Japan)

    2011-05-15

    Introduction: Glycine transporter 1 (GlyT-1) is an attractive target in positron emission tomography (PET) studies. Here, we report the in vivo evaluation of three carbon-11-labelled non-sarcosine-type GlyT-1 inhibitors - [{sup 11}C]SA1, [{sup 11}C]SA2 and [{sup 11}C]SA3 - as novel PET tracers for GlyT-1. Methods: The regional brain distributions of the three compounds in mice were studied at baseline and under receptor-blockade conditions with co-injection of carrier loading or pretreatment with an excess of selective GlyT-1 inhibitors (ALX-5407 and SSR504734). Metabolic stability was investigated by radio high-performance liquid chromatography. Dynamic PET scans in conscious monkeys were performed with/without selective GlyT-1 inhibitors. Results: The IC{sub 50} values of SA1, SA2 and SA3 were 9.0, 6400 and 39.7 nM, respectively. The regional brain uptakes of [{sup 11}C]SA1 and [{sup 11}C]SA3 in mice were heterogeneous and consistent with the known distribution of GlyT-1. [{sup 11}C]SA2 showed low and homogeneous uptake in the brain. Most radioactivity in the brain was detected in unchanged form, although peripherally these compounds were degraded. Carrier loading decreased the uptake of [{sup 11}C]SA1 in GlyT-1-rich regions. However, similar reductions were not observed with [{sup 11}C]SA3. Pretreatment with ALX-5407 decreased the uptake of [{sup 11}C]SA1 in GlyT-1-rich regions. In the monkey at baseline, regional brain uptake of [{sup 11}C]SA1 was heterogeneous and consistent with the known GlyT-1 distribution. Pretreatment with selective GlyT-1 inhibitors significantly decreased the distribution volume ratio of [{sup 11}C] SA1 in GlyT-1-rich regions. Conclusions: [{sup 11}C]SA1 has the most suitable profile among the three carbon-11-labelled GlyT-1 inhibitors. Lead optimization of [{sup 11}C]SA1 structure will be required to achieve in vivo selective GlyT-1 imaging.

  19. In vivo evaluation of carbon-11-labelled non-sarcosine-based glycine transporter 1 inhibitors in mice and conscious monkeys

    Toyohara, Jun; Ishiwata, Kiichi; Sakata, Muneyuki; Wu, Jin; Nishiyama, Shingo; Tsukada, Hideo; Hashimoto, Kenji

    2011-01-01

    Introduction: Glycine transporter 1 (GlyT-1) is an attractive target in positron emission tomography (PET) studies. Here, we report the in vivo evaluation of three carbon-11-labelled non-sarcosine-type GlyT-1 inhibitors - [ 11 C]SA1, [ 11 C]SA2 and [ 11 C]SA3 - as novel PET tracers for GlyT-1. Methods: The regional brain distributions of the three compounds in mice were studied at baseline and under receptor-blockade conditions with co-injection of carrier loading or pretreatment with an excess of selective GlyT-1 inhibitors (ALX-5407 and SSR504734). Metabolic stability was investigated by radio high-performance liquid chromatography. Dynamic PET scans in conscious monkeys were performed with/without selective GlyT-1 inhibitors. Results: The IC 50 values of SA1, SA2 and SA3 were 9.0, 6400 and 39.7 nM, respectively. The regional brain uptakes of [ 11 C]SA1 and [ 11 C]SA3 in mice were heterogeneous and consistent with the known distribution of GlyT-1. [ 11 C]SA2 showed low and homogeneous uptake in the brain. Most radioactivity in the brain was detected in unchanged form, although peripherally these compounds were degraded. Carrier loading decreased the uptake of [ 11 C]SA1 in GlyT-1-rich regions. However, similar reductions were not observed with [ 11 C]SA3. Pretreatment with ALX-5407 decreased the uptake of [ 11 C]SA1 in GlyT-1-rich regions. In the monkey at baseline, regional brain uptake of [ 11 C]SA1 was heterogeneous and consistent with the known GlyT-1 distribution. Pretreatment with selective GlyT-1 inhibitors significantly decreased the distribution volume ratio of [ 11 C] SA1 in GlyT-1-rich regions. Conclusions: [ 11 C]SA1 has the most suitable profile among the three carbon-11-labelled GlyT-1 inhibitors. Lead optimization of [ 11 C]SA1 structure will be required to achieve in vivo selective GlyT-1 imaging.

  20. Functional characterisation of the human alpha1 glycine receptor in a fluorescence-based membrane potential assay

    Jensen, Anders A.; Kristiansen, Uffe

    2004-01-01

    In the present study, we have created a stable HEK293 cell line expressing the human homomeric alpha1 glycine receptor (GlyR) and characterised its functional pharmacology in a conventional patch-clamp assay and in the FLIPR Membrane Potential (FMP) assay, a fluorescence-based high throughput scr...... not be suited for sophisticated studies of GlyR pharmacology and kinetics. However, the assay offers several advantages in studies of ligand-receptor interactions. Furthermore, the assay could be highly useful in the search for structurally novel ligands acting at GlyRs.......In the present study, we have created a stable HEK293 cell line expressing the human homomeric alpha1 glycine receptor (GlyR) and characterised its functional pharmacology in a conventional patch-clamp assay and in the FLIPR Membrane Potential (FMP) assay, a fluorescence-based high throughput...... ion did not appear to potentiate GlyR function at lower concentrations. Analogously, whereas pregnenolone sulphate inhibited alpha1 GlyR function, the potentiation of alpha1 GlyR by pregnenolone in electrophysiological studies could not be reproduced in the assay. In conclusion, the FMP assay may...

  1. Labeling of amino acids and peptides with isotopic oxygen as followed by 17O-N.M.R

    Steinschneider, A.; Burgar, M.I.; Buku, A.; Fiat, D.

    1981-01-01

    17 O was introduced into the respective α- and γ-COOH groups of Boc-Gly and Boc-Glu by saponification of the corresponding O-methyl esters with 1 N NaOH in H 2 17 O. Other 17 O enriched Boc-amino acids were prepared by acid catalyzed exchange into the amino acid α-COOH group followed by t-butyloxycarbonylation with t-butyl S-4, 6-dimethylpyrimidin-2-ylthio carbonate. Final enrichment, by approximately three orders of magnitude over natural abundance, was 60-100% of the possible maximum. The synthesis of [ 17 O]-Gly-Ala, [ 17 O]-Gly-Leu and [ 17 O]-Gly-Glu by DCC/HBT mediated coupling of Boc-Gly-[ 17 O]-α-COOH with amino acid-O-t-butyl esters followed by deprotection with HCL/EtOAc proceeded without undue loss of the isotope. Boc-[ 17 O]-Pro-Leu-Gly-NH 2 was prepared by a similar procedure. [Tyr 2 - 17 O]-, Pro 7 - 17 O]- and [Gly 4 - 17 O]-oxytocin were synthesized using solid phase support. 17 O-chemical shifts of synthetic intermediates and of the final products were as expected for each functional group. Linewith data correlate with the molecular weights of the compounds prepared. (author)

  2. Electrophysiological evidence of increased glycine receptor-mediated phasic and tonic inhibition by blockade of glycine transporters in spinal superficial dorsal horn neurons of adult mice

    Misa Oyama

    2017-03-01

    Full Text Available To understand the synaptic and/or extrasynaptic mechanisms underlying pain relief by blockade of glycine transporter subtypes GlyT1 and GlyT2, whole-cell recordings were made from dorsal horn neurons in spinal slices from adult mice, and the effects of NFPS and ALX-1393, selective GlyT1 and GlyT2 inhibitors, respectively, on phasic evoked or miniature glycinergic inhibitory postsynaptic currents (eIPSCs or mIPSCs were examined. NFPS and ALX-1393 prolonged the decay phase of eIPSCs without affecting their amplitude. In the presence of tetrodotoxin to record mIPSCs, NFPS and ALX-1393 induced a tonic inward current that was reversed by strychnine. Although NFPS had no statistically significant influences on mIPSCs, ALX-1393 significantly increased their frequency. We then further explored the role of GlyTs in the maintenance of glycinergic IPSCs. To facilitate vesicular release of glycine, repetitive high-frequency stimulation (HFS was applied at 10 Hz for 3 min during continuous recordings of eIPSCs at 0.1 Hz. Prominent suppression of eIPSCs was evident after HFS in the presence of ALX-1393, but not NFPS. Thus, it appears that phasic and tonic inhibition may contribute to the analgesic effects of GlyT inhibitors. However, reduced glycinergic inhibition due to impaired vesicular refilling could hamper the analgesic efficacy of GlyT2 inhibitors.

  3. Intermolecular Interactions in Ternary Glycerol–Sample–H2O

    Westh, Peter; Rasmussen, Erik Lumby; Koga, Yoshikata

    2011-01-01

    We studied the intermolecular interactions in ternary glycerol (Gly)–sample (S)–H2O systems at 25 °C. By measuring the excess partial molar enthalpy of Gly, HGlyEHEGly, we evaluated the Gly–Gly enthalpic interaction, HGly-GlyEHEGly--Gly, in the presence of various samples (S). For S, tert...... little effect on HGly-GlyEHEGly--Gly. This contrasts with our earlier studies on 1P–S–H2O in that Na+, F− and Cl− are found as hydration centers from the induced changes on HIP-IPEHEIP--IP in the presence of S, while Br−, I−, and SCN− are found to act as hydrophiles. In comparison with the Hofmeister...... ranking of these ions, the kosmotropes are hydration centers and the more kosmotropic the higher the hydration number, consistent with the original Hofmeister’s concept of “H2O withdrawing power.” Br−, I− and SCN−, on the other hand, acted as hydrophiles and the more chaotropic they are the more...

  4. A critical role for glycine transporters in hyperexcitability disorders

    Robert J Harvey

    2008-03-01

    Full Text Available Defects in mammalian glycinergic neurotransmission result in a complex motor disorder characterized by neonatal hypertonia and an exaggerated startle refl ex, known as hyperekplexia (OMIM 149400. This affects newborn children and is characterized by noise or touch-induced seizures that result in muscle stiffness and breath-holding episodes. Although rare, this disorder can have serious consequences, including brain damage and/or sudden infant death. The primary cause of hyperekplexia is missense and nonsense mutations in the glycine receptor (GlyR α1 subunit gene (GLRA1 on chromosome 5q33.1, although we have also discovered rare mutations in the genes encoding the GlyR β subunit (GLRB and the GlyR clustering proteins gephyrin (GPNH and collybistin (ARHGEF9. Recent studies of the Na+ /Cl--dependent glycine transporters GlyT1 and GlyT2 using mouse knockout models and human genetics have revealed that mutations in GlyT2 are a second major cause of hyperekplexia, while the phenotype of the GlyT1 knockout mouse resembles a devastating neurological disorder known as glycine encephalopathy (OMIM 605899. These findings highlight the importance of these transporters in regulating the levels of synaptic glycine.

  5. Glycine-extended gastrin enhances somatostatin release from cultured rabbit fundic D-cells [v1; ref status: indexed, http://f1000r.es/8n

    Ian LP Beales

    2013-02-01

    Full Text Available The role of the peptide hormone gastrin in stimulating gastric acid secretion is well established. Mature amidated gastrin is processed from larger peptide precursor forms. Increasingly these processing intermediates, such as glycine-extended gastrin (G-Gly and progastrin, have been shown to have biological activities of their own, often separate and complementary to gastrin. Although G-Gly is synthesized and secreted by gastric antral G-cells, the physiological functions of this putative mediator are unclear. Gastrin and cholecystokinin (CCK stimulate the secretion of somatostatin from gastric D-cells as part of the feedback control of gastric acid. In this study the effect of G-Gly and gastrin on the release of somatostatin from rabbit fundic D-cells was examined. D-cells were obtained by collagenase-EDTA digestion and elutriation and cultured for 48 hours. With a 2 hour exposure to the peptides, gastrin but not G-Gly stimulated somatostatin release. Treatment of D-cells for 24 hours with gastrin or G-Gly individually, significantly enhanced subsequent basal as well as CCK- and GLP-1-stimulated somatostatin release. Twenty four hours exposure to gastrin combined with G-Gly synergistically enhanced basal and agonist-stimulated somatostatin release and cellular somatostatin content. Gastrin and G-Gly may be important in the longer term regulation of D-cell function.

  6. Taurolidine as an effective and biocompatible additive for plaque-removing techniques on implant surfaces.

    John, Gordon; Schwarz, Frank; Becker, Jürgen

    2015-06-01

    The aim of the present study was the evaluation of the effectiveness and efficiency of two plaque-removing techniques, plastic curettes (PC) and glycine powder airflow (GLY) in combination with taurolidine (T), chlorhexidine (CHX), or pure water (PW) as additives and compared to groups without previous treatment (NT). Plaque was collected on titanium samples for 48 h in six subjects. Specimens were worn in a special splint in the upper jaw and randomly assigned to test and control groups. After biofilm removal procedures, clean implant surface (CIS) on the samples and treatment time were taken as parameters. Mean CIS was determined in the following descending order: T-GLY > CHX-GLY > NT-GLY > T-PC > PW-GLY > PW-PC > CHX-PC > NT-PC. Mean treatment time was determined in the following ascending order: T-GLY treatment times of the T groups were significantly lower than their corresponding PC or GLY groups. The results of the current study indicate that taurolidine seems to enhance effectiveness of plaque-removing procedures with plastic curettes and glycine powder airflow. Also, the efficiency of both treatment procedures seems to be increased.

  7. Experiences from treatment-predictive KRAS testing; high mutation frequency in rectal cancers from females and concurrent mutations in the same tumor

    Jönsson, Mats; Ekstrand, Anna; Edekling, Thomas

    2009-01-01

    . METHODS: We used a real-time PCR based method to determine KRAS mutations in 136 colorectal cancers with mutations identified in 53 (39%) tumors. RESULTS: KRAS mutations were significantly more often found in rectal cancer (21/38, 55%) than in colon cancer (32/98, 33%) (P = 0.02). This finding...... was explained by marked differences mutation rates in female patients who showed mutations in 33% of the colon cancers and in 67% of the rectal cancers (P = 0.01). Concurrent KRAS mutations were identified in three tumors; two colorectal cancers harbored Gly12Asp/Gly13Asp and Gly12Cys/Gly13Asp and a third tumor...... carried Gly12Cys/Gly12Asp in an adenomatous component and additionally acquired Gly12Val in the invasive component. CONCLUSION: The demonstration of a particularly high KRAS mutation frequency among female rectal cancer patients suggests that this subset is the least likely to respond to anti...

  8. Evaluation of a FRET-peptide substrate to predict virulence in Pseudomonas aeruginosa.

    Wendy E Kaman

    Full Text Available Pseudomonas aeruginosa produces a number of proteases that are associated with virulence and disease progression. A substrate able to detect P. aeruginosa-specific proteolytic activity could help to rapidly alert clinicians to the virulence potential of individual P. aeruginosa strains. For this purpose we designed a set of P. aeruginosa-specific fluorogenic substrates, comprising fluorescence resonance energy transfer (FRET-labeled peptides, and evaluated their applicability to P. aeruginosa virulence in a range of clinical isolates. A FRET-peptide comprising three glycines (3xGly was found to be specific for the detection of P. aeruginosa proteases. Further screening of 97 P. aeruginosa clinical isolates showed a wide variation in 3xGly cleavage activity. The absence of 3xGly degradation by a lasI knock out strain indicated that 3xGly cleavage by P. aeruginosa could be quorum sensing (QS-related, a hypothesis strengthened by the observation of a strong correlation between 3xGly cleavage, LasA staphylolytic activity and pyocyanin production. Additionally, isolates able to cleave 3xGly were more susceptible to the QS inhibiting antibiotic azithromycin (AZM. In conclusion, we designed and evaluated a 3xGly substrate possibly useful as a simple tool to predict virulence and AZM susceptibility.

  9. Association of expression of the hedgehog signal with Merkel cell polyomavirus infection and prognosis of Merkel cell carcinoma.

    Kuromi, Teruyuki; Matsushita, Michiko; Iwasaki, Takeshi; Nonaka, Daisuke; Kuwamoto, Satoshi; Nagata, Keiko; Kato, Masako; Akizuki, Gen; Kitamura, Yukisato; Hayashi, Kazuhiko

    2017-11-01

    Merkel cell carcinoma (MCC) is an aggressive neuroendocrine skin cancer that mostly occurs in the elderly. Merkel cell polyomavirus (MCPyV) is detected in approximately 80% of MCCs and is associated with carcinogenesis. Hedgehog signaling pathway plays a role in human embryogenesis and organogenesis. In addition, reactivation of this pathway later in life can cause tumors. Twenty-nineMCPyV-positive and 21 MCPyV-negative MCCs were immunohistochemically stained with primary antibodies for hedgehog signaling (SHH, IHH, PTCH1, SMO, GLI1, GLI2, and GLI3) and evaluated using H-score. Polymerase chain reaction and sequence analysis for SHH and GLI1 exons were also performed. Expression of SHH was higher in MCPyV-positive MCCs than in MCPyV-negative MCCs (PA. Only 2 mutations with amino acid changes were detected in MCPyV-negative MCCs only: 1 missense mutation in GLI1 exon 4 and 1 nonsense mutation in SHH-3B. Expression of SHH and GLI1 may be useful prognostic markers of MCC because increased expression was associated with better prognosis. The high rate of c.576G>A silent mutation in GLI1 exon 5 was a feature of MCC. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. Glycine Receptor Activation Impairs ATP-Induced Calcium Transients in Cultured Cortical Astrocytes

    Tatiana P. Morais

    2018-01-01

    Full Text Available In central nervous system, glycine receptor (GlyR is mostly expressed in the spinal cord and brainstem, but glycinergic transmission related elements have also been identified in the brain. Astrocytes are active elements at the tripartite synapse, being responsible for the maintenance of brain homeostasis and for the fine-tuning of synaptic activity. These cells communicate, spontaneously or in response to a stimulus, by elevations in their cytosolic calcium (calcium transients, Ca2+T that can be propagated to other cells. How these Ca2+T are negatively modulated is yet poorly understood. In this work, we evaluated GlyR expression and its role on calcium signaling modulation in rat brain astrocytes. We first proved that GlyR, predominantly subunits α2 and β, was expressed in brain astrocytes and its localization was confirmed in the cytoplasm and astrocytic processes by immunohistochemistry assays. Calcium imaging experiments in cultured astrocytes showed that glycine (500 μM, a GlyR agonist, caused a concentration-dependent reduction in ATP-induced Ca2+T, an effect abolished by the GlyR antagonist, strychnine (0.8 μM, as well as by nocodazole (1 μM, known to impair GlyR anchorage to the plasma membrane. This effect was mimicked by activation of GABAAR, another Cl--permeable channel. In summary, we demonstrated that GlyR activation in astrocytes mediates an inhibitory effect upon ATP induced Ca2+T, which most probably involves changes in membrane permeability to Cl- and requires GlyR anchorage at the plasma membrane. GlyR in astrocytes may thus be part of a mechanism to modulate astrocyte-to-neuron communication.

  11. Measurement of endogenous allergens in genetically modified soybeans--short communication.

    Ladics, Gregory S; Budziszewski, Gregory J; Herman, Rod A; Herouet-Guicheney, Corinne; Joshi, Saurabh; Lipscomb, Elizabeth A; McClain, Scott; Ward, Jason M

    2014-10-01

    The measurement of endogenous allergens is required by the European Commission (EC) as part of the compositional analysis for GM products from host plants that are common causes of food allergy, such as soybean (EC Implementing Regulation No. 503/2013). In each case, the EC Implementing Regulation indicates that analysis be conducted on identified allergens as specified in the Organization of Economic Cooperation and Development (OECD) consensus documents on compositional considerations for new plant varieties. This communication discusses the methods available to measure endogenous allergens as well as the endogenous soybean allergens that should be analyzed. It is suggested herein that in conjunction with the 2012 OECD consensus document on soybean, any list of soybean allergens should be based on clinically relevant data among publicly available allergen databases and peer-reviewed scientific publications, and the ability to measure the identified allergen. Based on a detailed analysis of the scientific literature, the following key points are recommended: (1) the acceptance of serum-free, quantitative analytical method data as an alternative to traditional IgE reactivity qualitative or semi-quantitative data for evaluation of endogenous soybean allergen content; (2) eight of the 15 potential allergens listed in the OECD soybean consensus document (Gly m 3, Gly m 4, Gly m Bd28K, Gly m Bd30K, Gly m 5, Gly m 6, Gly m 8, and Kunitz trypsin inhibitor) have both appropriate supporting clinical data and sufficient sequence information to be evaluated in comparative endogenous soybean allergen studies; and (3) the remaining seven proteins (Gly m 1, Gly m 2, unknown 50kDa protein, unknown 39kDa protein, P-22-25, lipoxygenase and lectin) lack sufficient data for clear classification as confirmed allergens and/or available sequence information and should not be currently included in the measurement of endogenous soybean allergens in the compositional analysis for the EU

  12. Voltage-Dependent Inhibition of Glycine Receptor Channels by Niflumic Acid

    Galyna Maleeva

    2017-05-01

    Full Text Available Niflumic acid (NFA is a member of the fenamate class of nonsteroidal anti-inflammatory drugs. This compound and its derivatives are used worldwide clinically for the relief of chronic and acute pain. NFA is also a commonly used blocker of voltage-gated chloride channels. Here we present evidence that NFA is an efficient blocker of chloride-permeable glycine receptors (GlyRs with subunit heterogeneity of action. Using the whole-cell configuration of patch-clamp recordings and molecular modeling, we analyzed the action of NFA on homomeric α1ΔIns, α2B, α3L, and heteromeric α1β and α2β GlyRs expressed in CHO cells. NFA inhibited glycine-induced currents in a voltage-dependent manner and its blocking potency in α2 and α3 GlyRs was higher than that in α1 GlyR. The Woodhull analysis suggests that NFA blocks α1 and α2 GlyRs at the fractional electrical distances of 0.16 and 0.65 from the external membrane surface, respectively. Thus, NFA binding site in α1 GlyR is closer to the external part of the membrane, while in α2 GlyR it is significantly deeper in the pore. Mutation G254A at the cytoplasmic part of the α1 GlyR pore-lining TM2 helix (level 2′ increased the NFA blocking potency, while incorporation of the β subunit did not have a significant effect. The Hill plot analysis suggests that α1 and α2 GlyRs are preferably blocked by two and one NFA molecules, respectively. Molecular modeling using Monte Carlo energy minimizations provides the structural rationale for the experimental data and proposes more than one interaction site along the pore where NFA can suppress the ion permeation.

  13. Mutation of a zinc-binding residue in the glycine receptor α1 subunit changes ethanol sensitivity in vitro and alcohol consumption in vivo.

    McCracken, Lindsay M; Blednov, Yuri A; Trudell, James R; Benavidez, Jillian M; Betz, Heinrich; Harris, R Adron

    2013-02-01

    Ethanol is a widely used drug, yet an understanding of its sites and mechanisms of action remains incomplete. Among the protein targets of ethanol are glycine receptors (GlyRs), which are potentiated by millimolar concentrations of ethanol. In addition, zinc ions also modulate GlyR function, and recent evidence suggests that physiologic concentrations of zinc enhance ethanol potentiation of GlyRs. Here, we first built a homology model of a zinc-bound GlyR using the D80 position as a coordination site for a zinc ion. Next, we investigated in vitro the effects of zinc on ethanol action at recombinant wild-type (WT) and mutant α1 GlyRs containing the D80A substitution, which eliminates zinc potentiation. At D80A GlyRs, the effects of 50 and 200 mM ethanol were reduced as compared with WT receptors. Also, in contrast to what was seen with WT GlyRs, neither adding nor chelating zinc changed the magnitude of ethanol enhancement of mutant D80A receptors. Next, we evaluated the in vivo effects of the D80A substitution by using heterozygous Glra1(D80A) knock-in (KI) mice. The KI mice showed decreased ethanol consumption and preference, and they displayed increased startle responses compared with their WT littermates. Other behavioral tests, including ethanol-induced motor incoordination and strychnine-induced convulsions, revealed no differences between the KI and WT mice. Together, our findings indicate that zinc is critical in determining the effects of ethanol at GlyRs and suggest that zinc binding at the D80 position may be important for mediating some of the behavioral effects of ethanol action at GlyRs.

  14. Un anello per domarli tutti

    Merlini, Antonio

    2014-01-01

    Questa tesina tratta degli acceleratori di particelle. I motivi della loro nascita, la loro evoluzione, il loro presente e il loro futuro. Particolare attenzione sarà data al Large Hadron Collider, il più grande di essi e la più incredibile macchina mai realizzata dall’uomo. Inoltre si parlerà dei rilevatori, che permettono lo studio di particelle invisibili all’uomo; del fatto che gli scienziati non escludono che gli strumenti che utilizzano possano potenzialmente distruggere il pianeta; e di come gli acceleratori hanno cambiato la nostra vita.

  15. An atlas of classification. Signage between open shelves, the Web and the catalogue

    Andrea Fabbrizzi

    2014-05-01

    Questa segnaletica è fondata sulla comunicazione cross-mediale, e integra le modalità comunicative della biblioteca a vari livelli, sia nel contesto dello stesso medium, sia tra media diversi: tra i cartelli sulle testate degli scaffali, tra questi cartelli e il sito web della biblioteca, tra il sito web e il catalogo. Per questo sistema integrato sono particolarmente adatti i dispositivi mobili come i tablet e gli smartphone, perché danno la possibilità di accedere al Web mentre ci si muove tra gli scaffali. Il collegamento diretto tra gli scaffali aperti classificati e il catalogo è reso possibile dai codici QR stampati sui cartelli.

  16. Elementi e relazioni in un sistema di classificazione dei record

    Maria Mata Caravaca

    2017-05-01

    Full Text Available La costruzione di schemi di classificazione dei record è un tema poco sviluppato a cui la disciplina archivistica ha finora dedicato poca attenzione. Non si tratta di un argomento di facile comprensione, dal momento che gli schemi di classificazione dei record sono gli strumenti principali utilizzati tradizionalmente per l'organizzazione e la gestione dei record. Questo articolo esamina gli elementi presi in considerazione nella costruzione di uno schema di classificazione, così come le modalità attraverso le quali questi elementi sono correlati. Vengono inoltre esaminate le relazioni gerarchiche e non gerearchiche, ed identificate le possibili future direzioni della ricerca su questi temi.

  17. JPRS Report, Science & Technology, USSR: Life Sciences.

    1988-02-12

    the [Leu]enkephalin pentapeptide Tyr-Gly-Gly-Phe-Leu (i)and its [D-Ala2]analog Tyr-D-Ala-Gly-Phe-Leu ( la ), which is more resistant to proteinases, by...Biology and Genetics, Ukrainian SSR Academy of Sciences, Kiev] ltl!tra°+V ^° la ^ion ^dies were conducted for mini forms of the M13 bacteri- ophage...of fodder substrates (hay, soya millet,,oats, wheat, barley, bean, etc.) Various degrees of growth were obtained on the various fodder products

  18. Effects of the Amino Acid Linkers on the Melanoma-Targeting and Pharmacokinetic Properties of Indium-111-labeled Lactam Bridge-Cyclized α-MSH Peptides

    Guo, Haixun; Yang, Jianquan; Gallazzi, Fabio; Miao, Yubin

    2011-01-01

    The purpose of this study was to examine the profound effects of the amino acid linkers on the melanoma targeting and pharmacokinetic properties of novel 111In-labeled lactam bridge-cyclized DOTA-[X]-CycMSHhex {1,4,7,10-Tetraazacyclododecane-1,4,7,10-tetraacetic acid-[X]-c[Asp-His-dPhe-Arg-Trp-Lys]-CONH2, X=GlyGlyNle, GlyGluNle or NleGlyGlu} peptides. Methods Three novel DOTA-GGNle-CycMSHhex, DOTA-GENle-CycMSHhex and DOTA-NleGE-CycMSHhex peptides were designed and synthesized. The melanocortin-1 (MC1) receptor binding affinities of the peptides were determined in B16/F1 melanoma cells. The melanoma targeting and pharmacokinetic properties of 111In-DOTA-GGNle-CycMSHhex and 111In-DOTA-GENle-CycMSHhex were determined in B16/F1 melanoma-bearing C57 mice. Results DOTA-GGNle-CycMSHhex and DOTA-GENle-CycMSHhex displayed 2.1 and 11.5 nM MC1 receptor binding affinities, whereas DOTA-NleGE-CycMSHhex showed 873.4 nM MC1 receptor binding affinity. The introduction of the -GlyGly- linker maintained high melanoma uptake while decreased the renal and liver uptakes of 111In-DOTA-GlyGlyNle-CycMSHhex. The tumor uptake values of 111In-DOTA-GGNle-CycMSHhex were 19.05 ± 5.04 and 18.6 ± 3.56 % injected dose/gram (%ID/g) at 2 and 4 h post-injection. 111In-DOTA-GGNle-CycMSHhex exhibited 28, 32 and 42% less renal uptake values than 111In-DOTA-Nle-CycMSHhex we reported previously, and 61, 65 and 68% less liver uptake values than 111In-DOTA-Nle-CycMSHhex at 2, 4 and 24 h post-injection, respectively. Conclusion The amino acid linkers exhibited the profound effects on the melanoma targeting and pharmacokinetic properties of the 111In-labeled lactam bridge-cyclized α-MSH peptides. Introduction of the -GlyGly- linker maintained high melanoma uptake while reducing the renal and liver uptakes of 111In-DOTA-GlyGlyNle-CycMSHhex, highlighting its potential as an effective imaging probe for melanoma detection, as well as a therapeutic peptide for melanoma treatment when labeled with a therapeutic

  19. Theoretical conformational analysis of the bovine adrenal medulla 12 residue peptide molecule

    Akhmedov, N. A.; Tagiyev, Z. H.; Hasanov, E. M.; Akverdieva, G. A.

    2003-02-01

    The spatial structure and conformational properties of the bovine adrenal medulla 12 residue peptide Tyr1-Gly2-Gly3-Phe4-Met5-Arg6-Arg7-Val8-Gly9-Arg10-Pro11-Glu12 (BAM-12P) molecule were studied by theoretical conformational analysis. It is revealed that this molecule can exist in several stable states. The energy and geometrical parameters for the low-energy conformations are obtained. The conformationally rigid and labile segments of this molecule were revealed.

  20. Recursive deconvolution of combinatorial chemical libraries.

    Erb, E; Janda, K D; Brenner, S

    1994-01-01

    A recursive strategy that solves for the active members of a chemical library is presented. A pentapeptide library with an alphabet of Gly, Leu, Phe, and Tyr (1024 members) was constructed on a solid support by the method of split synthesis. One member of this library (NH2-Tyr-Gly-Gly-Phe-Leu) is a native binder to a beta-endorphin antibody. A variation of the split synthesis approach is used to build the combinatorial library. In four vials, a member of the library's alphabet is coupled to a...