Telomerase activity in the vaginal margins of radical hysterectomy in patients with carcinoma of the cervix: Correlation with histology and human papillomavirus

This study was undertaken to evaluate the telomerase activity both in the tumor and in the vaginal margins of radical hysterectomy in patients with squamous cell carcinoma (SCC) of the cervix. Thirty-three patients with SCC of the cervix (study group) and 13 patients with uterine myoma (control group) were prospectively studied. Tissue samples were taken from the tumor or cervix, anterior vaginal margin (AVM), and posterior vaginal margin (PVM). The specimens were analyzed by histopathology, by a telomerase PCR-TRAP-ELISA kit, and by polymerase chain reaction using human papillomavirus (HPV) DNA. The telomerase activity was significantly higher in the tumor than in the benign cervix (P < 0.001). There was no difference in telomerase activity in the AVM and PVM in patients with cervical carcinoma compared to the control group. Telomerase activity was associated with the presence of histologic malignancy in the PVM of patients submitted to radical hysterectomy (P = 0.03). This association was not observed with the presence of HPV in AVM or PVM in the study group. Telomerase activity is a marker of histologic malignancy in patients with SCC of the cervix. There was no association between the telomerase activity and the presence of HPV in vaginal margins of patients submitted to radical hysterectomy. © 2006, Copyright the Authors.

Rastreabilidade da farinha de vísceras de aves em codornas submetidas a longo período de criação utilizando a técnica dos isótopos estáveis 'delta' 'intpot. 13 c' e 'delta' 'intpot. 15 N'

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Modeling the spatial dynamics of economic models

The advances that have been characterizing spatial econometrics in recent years are mostly theoretical and have not found an extensive empirical application yet. In this work we aim at supplying a review of the main tools of spatial econometrics and to show an empirical application for one of the most recently introduced estimators. Despite the numerous alternatives that the econometric theory provides for the treatment of spatial (and spatiotemporal) data, empirical analyses are still limited by the lack of availability of the correspondent routines in statistical and econometric software. Spatiotemporal modeling represents one of the most recent developments in spatial econometric theory and the finite sample properties of the estimators that have been proposed are currently being tested in the literature. We provide a comparison between some estimators (a quasi-maximum likelihood, QML, estimator and some GMM-type estimators) for a fixed effects dynamic panel data model under certain conditions, by means of a Monte Carlo simulation analysis. We focus on different settings, which are characterized either by fully stable or quasi-unit root series. We also investigate the extent of the bias that is caused by a non-spatial estimation of a model when the data are characterized by different degrees of spatial dependence. Finally, we provide an empirical application of a QML estimator for a time-space dynamic model which includes a temporal, a spatial and a spatiotemporal lag of the dependent variable. This is done by choosing a relevant and prolific field of analysis, in which spatial econometrics has only found limited space so far, in order to explore the value-added of considering the spatial dimension of the data. In particular, we study the determinants of cropland value in Midwestern U.S.A. in the years 1971-2009, by taking the present value model (PVM) as the theoretical framework of analysis.

Molecular cloning and characterization of NcROP2Fam-1, a member of the ROP2 family of rhoptry proteins in Neospora caninum that is targeted by antibodies neutralizing host cell invasion in vitro.

Recent publications demonstrated that a fragment of a Neospora caninum ROP2 family member antigen represents a promising vaccine candidate. We here report on the cloning of the cDNA encoding this protein, N. caninum ROP2 family member 1 (NcROP2Fam-1), its molecular characterization and localization. The protein possesses the hallmarks of ROP2 family members and is apparently devoid of catalytic activity. NcROP2Fam-1 is synthesized as a pre-pro-protein that is matured to 2 proteins of 49 and 55 kDa that localize to rhoptry bulbs. Upon invasion the protein is associated with the nascent parasitophorous vacuole membrane (PVM), evacuoles surrounding the host cell nucleus and, in some instances, the surface of intracellular parasites. Staining was also observed within the cyst wall of 'cysts' produced in vitro. Interestingly, NcROP2Fam-1 was also detected on the surface of extracellular parasites entering the host cells and antibodies directed against NcROP2Fam-1-specific peptides partially neutralized invasion in vitro. We conclude that, in spite of the general belief that ROP2 family proteins are intracellular antigens, NcROP2Fam-1 can also be considered as an extracellular antigen, a property that should be taken into account in further experiments employing ROP2 family proteins as vaccines.

A Plasmodium phospholipase is involved in disruption of the liver stage parasitophorous vacuole membrane

The coordinated exit of intracellular pathogens from host cells is a process critical to the success and spread of an infection. While phospholipases have been shown to play important roles in bacteria host cell egress and virulence, their role in the release of intracellular eukaryotic parasites is largely unknown. We examined a malaria parasite protein with phospholipase activity and found it to be involved in hepatocyte egress. In hepatocytes, Plasmodium parasites are surrounded by a parasitophorous vacuole membrane (PVM), which must be disrupted before parasites are released into the blood. However, on a molecular basis, little is known about how the PVM is ruptured. We show that Plasmodium berghei phospholipase, PbPL, localizes to the PVM in infected hepatocytes. We provide evidence that parasites lacking PbPL undergo completely normal liver stage development until merozoites are produced but have a defect in egress from host hepatocytes. To investigate this further, we established a live-cell imaging-based assay, which enabled us to study the temporal dynamics of PVM rupture on a quantitative basis. Using this assay we could show that PbPL-deficient parasites exhibit impaired PVM rupture, resulting in delayed parasite egress. A wild-type phenotype could be re-established by gene complementation, demonstrating the specificity of the PbPL deletion phenotype. In conclusion, we have identified for the first time a Plasmodium phospholipase that is important for PVM rupture and in turn for parasite exit from the infected hepatocyte and therefore established a key role of a parasite phospholipase in egress.

Long-term live imaging reveals cytosolic immune responses of host hepatocytes against Plasmodium infection and parasite escape mechanisms

Plasmodium parasites are transmitted by Anopheles mosquitoes to the mammalian host and actively infect hepatocytes after passive transport in the bloodstream to the liver. In their target host hepatocyte, parasites reside within a parasitophorous vacuole (PV). In the present study it was shown that the parasitophorous vacuole membrane (PVM) can be targeted by autophagy marker proteins LC3, ubiquitin, and SQSTM1/p62 as well as by lysosomes in a process resembling selective autophagy. The dynamics of autophagy marker proteins in individual Plasmodium berghei-infected hepatocytes were followed by live imaging throughout the entire development of the parasite in the liver. Although the host cell very efficiently recognized the invading parasite in its vacuole, the majority of parasites survived this initial attack. Successful parasite development correlated with the gradual loss of all analyzed autophagy marker proteins and associated lysosomes from the PVM. However, other autophagic events like nonselective canonical autophagy in the host cell continued. This was indicated as LC3, although not labeling the PVM anymore, still localized to autophagosomes in the infected host cell. It appears that growing parasites even benefit from this form of nonselective host cell autophagy as an additional source of nutrients, as in host cells deficient for autophagy, parasite growth was retarded and could partly be rescued by the supply of additional amino acid in the medium. Importantly, mouse infections with P. berghei sporozoites confirmed LC3 dynamics, the positive effect of autophagy activation on parasite growth, and negative effects upon autophagy inhibition.

Transmembrane Insertion of the Toxoplasma gondii GRA5 Protein Occurs after Soluble Secretion into the Host Cell

The intracellular parasite Toxoplasma gondii resides within a specialized compartment, the parasitophorous vacuole (PV), that resists fusion with host cell endocytic and lysosomal compartments. The PV is extensively modified by secretion of parasite proteins, including the dense granule protein GRA5 that is specifically targeted to the delimiting membrane of the PV (PVM). We show here that GRA5 is present both in a soluble form and in hydrophobic aggregates. GRA5 is secreted as a soluble form into the PV after which it becomes stably associated with the PVM. Topological studies demonstrated that GRA5 was inserted into the PVM as a transmembrane protein with its N-terminal domain extending into the cytoplasm and its C terminus in the vacuole lumen. Deletion of 8 of the 18 hydrophobic amino acids of the single predicted transmembrane domain resulted in the failure of GRA5 to associate with the PVM; yet it remained correctly packaged in the dense granules and was secreted as a soluble protein into the PV. Collectively, these studies demonstrate that the secretory pathway in Toxoplasma is unusual in two regards; it allows soluble export of proteins containing typical transmembrane domains and provides a mechanism for their insertion into a host cell membrane after secretion from the parasite.

Toxoplasma invasion: the parasitophorous vacuole is formed from host cell plasma membrane and pinches off via a fission pore.

Most intracellular pathogens avoid lysing their host cells during invasion by wrapping themselves in a vacuolar membrane. This parasitophorous vacuole membrane (PVM) is often retained, serving as a critical transport interface between the parasite and the host cell cytoplasm. To test whether the PVM formed by the parasite Toxoplasma gondii is derived from host cell membrane or from lipids secreted by the parasite, we used time-resolved capacitance measurements and video microscopy to assay host cell surface area during invasion. We observed no significant change in host cell surface area during PVM formation, demonstrating that the PVM consists primarily of invaginated host cell membrane. Pinching off of the PVM from the host cell membrane occurred after an unexpected delay (34-305 sec) and was seen as a 0.219 +/- 0.006 pF drop in capacitance, which corresponds well to the predicted surface area of the entire PVM (30-33 microns2). The formation and closure of a fission pore connecting the extracellular medium and the vacuolar space was detected as the PVM pinched off. This final stage of parasite entry was accomplished without any breach in cell membrane integrity.

The Empirical Content of the Present Value Model: A survey of the instrumental uses of farmland prices. Factor Markets Working Document No. 53, June 2013

After reviewing the Present Value Model (PVM), in its basic form and with its major extensions, the authors carried out a literature review on the instrumental uses of farm land prices; namely what land prices may reveal in the framework of the PVM. Urban influence, non-market goods and climate change are topics where the PVM used with applied data may reveal farmers’ or landowners’ beliefs or subjective values, which are discussed in this paper. There is also extensive discussion of the topic of public regulations, and how they may affect land price directly, or through its present value.

Enforcement and public information strategies for DWI general deterrence: the Boise City, Idaho experience. Final report.

National Highway Traffic Safety Administration, Office of Driver and Pedestrian Research, Washington, D.C.

An evaluation of a community service sanction for DWI: the Baton Rouge community service work program. Volume II. Final report.

National Highway Traffic Safety Administration, Washington, D.C.

The history of Scotland during the reigns of Queen Mary and of King James VI. till his accession to the crown of England. : With a review of Scottish history previous to that period; and an appendix containing original papers. ; In two volumes. /

Includes index.

Design and implementation of a windows-based parallel computing environment for large scale optimization

A parallel computing environment to support optimization of large-scale engineering systems is designed and implemented on Windows-based personal computer networks, using the master-worker model and the Parallel Virtual Machine (PVM). It is involved in decomposition of a large engineering system into a number of smaller subsystems optimized in parallel on worker nodes and coordination of subsystem optimization results on the master node. The environment consists of six functional modules, i.e. the master control, the optimization model generator, the optimizer, the data manager, the monitor, and the post processor. Object-oriented design of these modules is presented. The environment supports steps from the generation of optimization models to the solution and the visualization on networks of computers. User-friendly graphical interfaces make it easy to define the problem, and monitor and steer the optimization process. It has been verified by an example of a large space truss optimization. (C) 2004 Elsevier Ltd. All rights reserved.

RESPONSABILIDADE SOCIAL, PREGAÇÃO E EDUCAÇÃO: TENSÕES MISSIOLÓGICAS NO PROJETO MISSIONÁRIO DA IGREJA METODISTA EM TERRAS BRASILEIRAS

Esta pesquisa, Responsabilidade Social, Pregação e Educação: tensões missiológicas no projeto missionário da Igreja Metodista em terras brasileiras, tem como objetivo analisar as ações missionárias da Igreja Metodista do período de sua implantação até o final do período referente ao 18° Concílio Geral (2006-2011). A análise resgatou a tríade Responsabilidade Social, Pregação e Educação como uma marca identitária do movimento wesleyano e que esse princípio missionário nem sempre foi seguido pelo metodismo brasileiro. Notou-se também que a tríade serviu, em momentos decisivos, como elemento norteador do projeto missionário. Isso é percebido no período da implantação, bem como na constituição do Plano para Vida e Missão (PVM). Em que pese que as ações missionárias no metodismo brasileiro nem sempre foram providas de uma reflexão teológica sistemática, a tríade Responsabilidade Social, Pregação e Educação sempre funcionou como eixos essenciais da missão e de forma indissociável. Não foram poucos os momentos em que o projeto missionário metodista enfrentou tensões na sua execução. O ímpeto pelo crescimento numérico e expansão geográfica, muitas vezes, ofuscou a fidelidade à tríade. Nessa trajetória de pesquisa está construída em quatro capítulos, nos quais se busca a superação de uma compreensão histórica equivocada e faz-se necessário que o projeto missionário assuma os elementos essenciais da tríade como marco regulatório da missão metodista. As repetitivas opções equivocadas não colaboram com a nova compreensão de missão. O resultado da pesquisa encoraja futuras análises para que o metodismo assuma o papel de agente de Deus na missio Dei e para que o seu envolvimento missionário mantenha fidelidade à tríade Responsabilidade Social, Pregação e Educação.

Esquemas de volúmenes finitos de alto orden: implementación en GPUs y aplicación a la simulación de flujos geofísicos

En esta tesis doctoral se exponen los fundamentos teóricos necesarios en el diseño de esquemas numéricos de volúmenes finitos para sistemas hiperbólicos no conservativos de una y dos dimensiones. Para el caso unidimensional se repasan los conceptos de esquema camino-conservativo y esquema bien equilibrado, así como la extensión de los esquemas numéricos a alto orden, basados en la reconstrucción de estados. En particular, se presentan los esquemas de tipo PVM (Polynomial Viscosity Matrix), así como diversos esquemas de limitadores de flujo que resultan de la extensión natural del método WAF, utilizando como base algunos esquemas de tipo PVM. Para el caso bidimensional se aborda el diseño de esquemas numéricos camino-conservativos y bien equilibrados de volúmenes finitos para sistemas hiperbólicos no conservativos y su extensión a alto orden, en particular se presenta una reconstrucción de estados de tercer orden compacta y que resulta de la combinación WENO de paraboloides y planos. 
 Se presenta además el desarrollo de métodos numéricos para el sistema de aguas someras bidimensional de una capa. En particular se definen esquemas de primer orden de tipo HLL y FORCE y su extensión a alto orden, un método de limitadores de flujo basado en el esquema HLL-WAF, así como su implementación en arquitecturas de tipo GPU, usando el entorno de programación CUDA. A continuación, se presenta un esquema numérico de orden uno para el sistema de aguas someras de una capa bidimensional en coordenadas esféricas (longitud/latitud), así como la extensión natural del método de limitadores de flujo presentado en el Capítulo 3 a este sistema. Finalmente, se presenta la validación del esquema de limitadores de flujo mediante la simulación de tsunamis reales, y la comparación con datos de campo.