A Novel Chemically Modified Curcumin Reduces Severity of Experimental Periodontal Disease in Rats: Initial Observations

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cytogenetic and molecular data demonstrate that the bryconinae (ostariophysi, bryconidae) species from southeastern Brazil form a phylogenetic and phylogeographic unit

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Cellular interference in craniofrontonasal syndrome: males mosaic for mutations in the X-linked EFNB1 gene are more severely affected than true hemizygotes

Craniofrontonasal syndrome (CFNS), an X-linked disorder caused by loss-of-function mutations of EFNB1, exhibits a paradoxical sex reversal in phenotypic severity: females characteristically have frontonasal dysplasia, craniosynostosis and additional minor malformations, but males are usually more mildly affected with hypertelorism as the only feature. X-inactivation is proposed to explain the more severe outcome in heterozygous females, as this leads to functional mosaicism for cells with differing expression of EPHRIN-B1, generating abnormal tissue boundaries-a process that cannot occur in hemizygous males. Apparently challenging this model, males occasionally present with a more severe female-like CFNS phenotype. We hypothesized that such individuals might be mosaic for EFNB1 mutations and investigated this possibility in multiple tissue samples from six sporadically presenting males. Using denaturing high performance liquid chromatography, massively parallel sequencing and multiplex-ligation-dependent probe amplification (MLPA) to increase sensitivity above standard dideoxy sequencing, we identified mosaic mutations of EFNB1 in all cases, comprising three missense changes, two gene deletions and a novel point mutation within the 5' untranslated region (UTR). Quantification by Pyrosequencing and MLPA demonstrated levels of mutant cells between 15 and 69%. The 5' UTR variant mutates the stop codon of a small upstream open reading frame that, using a dual-luciferase reporter construct, was demonstrated to exacerbate interference with translation of the wild-type protein. These results demonstrate a more severe outcome in mosaic than in constitutionally deficient males in an X-linked dominant disorder and provide further support for the cellular interference mechanism, normally related to X-inactivation in females.

Coastal flood vulnerability assessment with geomatic methods: Test sites of western Thailand, Sydney (Australia) and aeolian islands (south tyrrhenian sea, Italy)

The work undertaken in this PhD thesis is aimed at the development and testing of an innovative methodology for the assessment of the vulnerability of coastal areas to marine catastrophic inundation (tsunami). Different approaches are used at different spatial scales and are applied to three different study areas: 1. The entire western coast of Thailand 2. Two selected coastal suburbs of Sydney – Australia 3. The Aeolian Islands, in the South Tyrrhenian Sea – Italy I have discussed each of these cases study in at least one scientific paper: one paper about the Thailand case study (Dall’Osso et al., in review-b), three papers about the Sydney applications (Dall’Osso et al., 2009a; Dall’Osso et al., 2009b; Dall’Osso and Dominey-Howes, in review) and one last paper about the work at the Aeolian Islands (Dall’Osso et al., in review-a). These publications represent the core of the present PhD thesis. The main topics dealt with are outlined and discussed in a general introduction while the overall conclusions are outlined in the last section.

Applicazione e confronto di modelli per lo studio della vulnerabilita da maremoto nel golfo di siracusa

Il lavoro svolto in questa tesi s’inserisce e sviluppa soprattutto nel campo dell’analisi della vulnerabilità relativa agli tsunami ed è centrato sull’analisi della vulnerabilità di strutture ed edifici. Per la precisione si è focalizzata l’attenzione su un’area geografica specifica, cioè si è considerata l’ipotesi che un maremoto colpisca le coste orientali della Sicilia ed in particolare della città di Siracusa. Questo lavoro di tesi prenderà in considerazione due modelli distinti per la stima della vulnerabilità: il modello SCHEMA (SCenarios for Hazard-induced Emergencies MAnagement) che prende il nome dal progetto europeo in cui è stato sviluppato e il modello PTVA (Papathoma Tsunami Vulnerability Assessment) introdotto da Papathoma et al. (2003) e successivamente modificato da Dominey-Howes et al. (2007) e da Dall’Osso et al. (2009). Tali modelli sono esempi dei due possibili approcci (quantitativo e qualitativo). Nei seguenti capitoli si sono trattate le curve di fragilità e di danno, in particolare seguendo la metodologia di Koshimura et al. (2009) ed il lavoro di Valencia et al. (2011). A seguire sono stati descritti i due metodi utilizzati per lo studio della vulnerabilità (SCHEMA, PTVA) ed il lavoro che è stato condotto nell’area di Siracusa. Il lavoro di tesi si è concluso mostrando i risultati della classificazione di vulnerabilità evidenziando e discutendo differenze e similarità delle mappe risultanti dai due metodi applicati.

Complement C3 is a novel plasma clot component with anti-fibrinolytic properties

Increased plasma clot density and prolonged lysis times are associated with cardiovascular disease. In this study, we employed a functional proteomics approach to identify novel clot components which may influence clot phenotypes.

Do invasive species perform better in their new ranges?

A fundamental assumption in invasion biology is that most invasive species exhibit enhanced performance in their introduced range relative to their home ranges. This idea has given rise to numerous hypotheses explaining “invasion success” by virtue of altered ecological and evolutionary pressures. There are surprisingly few data, however, testing the underlying assumption that the performance of introduced populations, including organism size, reproductive output, and abundance, is enhanced in their introduced compared to their native range. Here, we combined data from published studies to test this hypothesis for 26 plant and 27 animal species that are considered to be invasive. On average, individuals of these 53 species were indeed larger, more fecund, and more abundant in their introduced ranges. The overall mean, however, belied significant variability among species, as roughly half of the investigated species (N = 27) performed similarly when compared to conspecific populations in their native range. Thus, although some invasive species are performing better in their new ranges, the pattern is not universal, and just as many are performing largely the same across ranges.

An updated synthesis of the observed and projected impacts of climate change on the chemical, physical and biological processes in the oceans

The 5th Assessment Report (AR5) of the Intergovernmental Panel on Climate Change (IPCC) states with very high certainty that anthropogenic emissions have caused measurable changes in the physical ocean environment. These changes are summarized with special focus on those that are predicted to have the strongest, most direct effects on ocean biological processes; namely, ocean warming and associated phenomena (including stratification and sea level rise) as well as deoxygenation and ocean acidification. The biological effects of these changes are then discussed for microbes (including phytoplankton), plants, animals, warm and cold-water corals, and ecosystems. The IPCC AR5 highlighted several areas related to both the physical and biological processes that required further research. As a rapidly developing field, there have been many pertinent studies published since the cut off dates for the AR5, which have increased our understanding of the processes at work. This study undertook an extensive review of recently published literature to update the findings of the AR5 and provide a synthesized review on the main issues facing future oceans. The level of detail provided in the AR5 and subsequent work provided a basis for constructing projections of the state of ocean ecosystems in 2100 under two the Representative Concentration Pathways RCP4.5 and 8.5. Finally the review highlights notable additions, clarifications and points of departure from AR5 provided by subsequent studies.

Contrasting futures for ocean and society from different anthropogenic CO2 emissions scenarios

The ocean moderates anthropogenic climate change at the cost of profound alterations of its physics, chemistry, ecology, and services. Here, we evaluate and compare the risks of impacts on marine and coastal ecosystems—and the goods and services they provide—for growing cumulative carbon emissions under two contrasting emissions scenarios. The current emissions trajectory would rapidly and significantly alter many ecosystems and the associated services on which humans heavily depend. A reduced emissions scenario—consistent with the Copenhagen Accord’s goal of a global temperature increase of less than 2°C—is much more favorable to the ocean but still substantially alters important marine ecosystems and associated goods and services. The management options to address ocean impacts narrow as the ocean warms and acidifies. Consequently, any new climate regime that fails to minimize ocean impacts would be incomplete and inadequate.

Quality standards for bone conduction implants.

CONCLUSION Bone conduction implants are useful in patients with conductive and mixed hearing loss for whom conventional surgery or hearing aids are no longer an option. They may also be used in patients affected by single-sided deafness. OBJECTIVES To establish a consensus on the quality standards required for centers willing to create a bone conduction implant program. METHOD To ensure a consistently high level of service and to provide patients with the best possible solution the members of the HEARRING network have established a set of quality standards for bone conduction implants. These standards constitute a realistic minimum attainable by all implant clinics and should be employed alongside current best practice guidelines. RESULTS Fifteen items are thoroughly analyzed. They include team structure, accommodation and clinical facilities, selection criteria, evaluation process, complete preoperative and surgical information, postoperative fitting and assessment, follow-up, device failure, clinical management, transfer of care and patient complaints.

Winter, Louise & Albert

Signatur des Originals: S 36/F11902

Evidence for keratinocyte stem cells in vitro: Long term engraftment and persistence of transgene expression from retrovirus-transduced keratinocytes

Epidermis is renewed by a population of stem cells that have been defined in vivo by slow turnover, label retention, position in the epidermis, and enrichment in β1 integrin, and in vitro by clonogenic growth, prolonged serial passage, and rapid adherence to extracellular matrix. The goal of this study is to determine whether clonogenic cells with long-term growth potential in vitro persist in vivo and give rise to a fully differentiated epidermis. Human keratinocytes were genetically labeled in culture by transduction with a retrovirus encoding the lacZ gene and grafted to athymic mice. Analysis of the cultures before grafting showed that 21.1–27.8% of clonogenic cells with the capacity for >30 generations were successfully transduced. In vivo, β-galactosidase (β-gal) positive cells participated in the formation of a fully differentiated epithelium and were detected throughout the 40-week postgraft period, initially as loosely scattered clusters and later as distinct vertical columns. Viable cells recovered from excised grafts were seeded at clonal densities and 23.3–33.3% of the colonies thus formed were β-gal positive. In addition, no evidence of transgene inactivation was obtained: all keratinocyte colonies recovered from grafted tissue that were β-gal negative also lacked the lacZ transgene. These results show that cells with long-term growth properties in vitro do indeed persist in vivo and form a fully differentiated epidermis, thereby exhibiting the properties of stem cells.

RAD51 supports spontaneous non-homologous recombination in mammalian cells, but not the corresponding process induced by topoisomerase inhibitors

The RAD51 protein has been shown to participate in homologous recombination by promoting ATP-dependent homologous pairing and strand transfer reactions. In the present study, we have investigated the possible involvement of RAD51 in non-homologous recombination. We demonstrate that overexpression of CgRAD51 enhances the frequency of spontaneous non-homologous recombination in the hprt gene of Chinese hamster cells. However, the rate of non-homologous recombination induced by the topoisomerase inhibitors campothecin and etoposide was not altered by overexpression of RAD51. These results indicate that the RAD51 protein may perform a function in connection with spontaneous non-homologous recombination that is not essential to or not rate-limiting for non-homologous recombination induced by camptothecin or etoposide. We discuss the possibility that the role played by RAD51 in non-homologous recombination observed here may not be linked to non-homologous end-joining.

Efficient pyrophosphorolysis by a hepatitis B virus polymerase may be a primer-unblocking mechanism

Effective antiviral agents are thought to inhibit hepatitis B virus (HBV) DNA synthesis irreversibly by chain termination because reverse transcriptases (RT) lack an exonucleolytic activity that can remove incorporated nucleotides. However, since the parameters governing this inhibition are poorly defined, fully delineating the catalytic mechanism of the HBV-RT promises to facilitate the development of antiviral drugs for treating chronic HBV infection. To this end, pyrophosphorolysis and pyrophosphate exchange, two nonhydrolytic RT activities that result in the removal of newly incorporated nucleotides, were characterized by using endogenous avian HBV replication complexes assembled in vivo. Although these activities are presumed to be physiologically irrelevant for every polymerase examined, the efficiency with which they are catalyzed by the avian HBV-RT strongly suggests that it is the first known polymerase to catalyze these reactions under replicative conditions. The ability to remove newly incorporated nucleotides during replication has important biological and clinical implications: these activities may serve a primer-unblocking function in vivo. Analysis of pyrophosphorolysis on chain-terminated DNA revealed that the potent anti-HBV drug β-l-(−)-2′,3′-dideoxy-3′-thiacytidine (3TC) was difficult to remove by pyrophosphorolysis, in contrast to ineffective chain terminators such as ddC. This disparity may account for the strong antiviral efficacy of 3TC versus that of ddC. The HBV-RT pyrophosphorolytic activity may therefore be a novel determinant of antiviral drug efficacy, and could serve as a target for future antiviral drug therapy. The strong inhibitory effect of cytoplasmic pyrophosphate concentrations on viral DNA synthesis may also partly account for the apparent slow rate of HBV genome replication.