Isolation of dengue 2 virus from a patient with central nervous system involvement (transverse myelitis)

A dengue fever case is described in a 58-year-old male patient with febrile illness and thrombocytopenia complicated by neurological involvement characterized by transverse myelitis followed by weakness of both legs and flaccid paralysis. Muscle strength was much diminished and bilateral areflexia was observed. Dengue 2 (DEN-2) virus was isolated and the patient sero-converted by hemagglutination-inhibition and IgM-ELISA tests. The RT-PCR test was positive to DEN-2 in acute phase serum and culture supernatant, but negative in the cerebrospinal fluid. After three weeks of hospitalization the patient was discharged. No other infectious agent was detected in the blood and cerebrospinal fluid samples. The patient had full recovery from paralysis six months after the onset of DEN-2 infection.

Development of a "green process" for the isolation of natural functional extracts with anti-cancer activity - Application of high-pressure technology

Dissertação para obtenção do Grau de Mestre em Engenharia Química e Bioquímica

Isolation and encapsulation of a natural colourant using green technologies

The use of natural pigments instead of synthetic colourants is receiving growing interest in the food industry. In this field, cactus pears (Opuntia spp.) have been identified to be a promising betalainic crops covering a wide coloured spectrum. The aim of this work was to develop adequate clean and mild methodologies for the isolation and encapsulation of betacyanins, from cactus pear fruits (Opuntia spp.). Firstly, two different emerging technologies, namely PLE (Pressurized Liquid Extraction) and HPCDAE (High Pressure Carbon Dioxide-Assisted Extraction), were exploited to isolation of betacyanins form cactus pear fruits. Different process conditions were tested for the maximum recovery of betacyanins. Results showed that highest extraction yields were achieved for HPCDAE and mass ratio of pressurized carbon dioxide vs. acidified water was the parameter that most affected the betacyanins extraction. At optimum conditions of HPCDAE, Opuntia spp. extract presented a total betacyanin content of 211 ± 10 mg/100 g whereas extracts obtained using conventional extraction, PLE in static and in dynamic mode presented a total betacyanin content of 85 ± 3, 191 ± 2 and 153 ± 5 mg/100 g, respectively. HPCDAE has proven to be a successful technology to extract betacyanins from Opuntia spp. fruits. Afterward, Supercritical Fluid Technology was exploited to develop lipidic particles of betalain-rich extract. A betacyanin-rich conventional extract was encapsulated by PGSS® (Particles from Gas Saturated Solutions) technique. Different process conditions were tested in order to model the encapsulation of betacyanins. The pressure had a negative effect on betacyanin encapsulation. Lower pressures leads to an increase in the betacyanin encapsulation. This effect was more pronounced at higher temperatures and lower equilibrium time. At these conditions, Opuntia spp. particles presented 64.4 ± 4.5 mg/100 g and high antioxidant capacity. When compared with the Opuntia spp. dried extract, lipidic particles contributed to a better homogenization of the pink colour after incorporation in ice cream.

Integrated data model and DSL modifications

Companies are increasingly more and more dependent on distributed web-based software systems to support their businesses. This increases the need to maintain and extend software systems with up-to-date new features. Thus, the development process to introduce new features usually needs to be swift and agile, and the supporting software evolution process needs to be safe, fast, and efficient. However, this is usually a difficult and challenging task for a developer due to the lack of support offered by programming environments, frameworks, and database management systems. Changes needed at the code level, database model, and the actual data contained in the database must be planned and developed together and executed in a synchronized way. Even under a careful development discipline, the impact of changing an application data model is hard to predict. The lifetime of an application comprises changes and updates designed and tested using data, which is usually far from the real, production, data. So, coding DDL and DML SQL scripts to update database schema and data, is the usual (and hard) approach taken by developers. Such manual approach is error prone and disconnected from the real data in production, because developers may not know the exact impact of their changes. This work aims to improve the maintenance process in the context of Agile Platform by Outsystems. Our goal is to design and implement new data-model evolution features that ensure a safe support for change and a sound migration process. Our solution includes impact analysis mechanisms targeting the data model and the data itself. This provides, to developers, a safe, simple, and guided evolution process.

Notification of the first isolation of Cacipacore virus in a human in the State of Rondônia, Brazil

Flavivirus is a genus of arthropod-transmitted viruses of the family Flaviviridae, and in Brazil, up to eleven different Flavivirus have been isolated. We collected blood from farmers in the municipality of Theobroma, which is located 320km from the City of Porto Velho, the former capital of the Brazilian State of Rondônia. For viral isolation, we used newborn mouse brain, followed by RT-PCR with specific universal Flavivirus primers. We obtained fragments 958bp and 800bp in length. Based on BLAST, these sequences were 91% similar to a sequence of Cacipacore virus.

Tracking error of exchange-traded funds: Evidence from the UK

This paper is mainly concerned with the tracking accuracy of Exchange Traded Funds (ETFs) listed on the London Stock Exchange (LSE) but also evaluates their performance and pricing efficiency. The findings show that ETFs offer virtually the same return but exhibit higher volatility than their benchmark. It seems that the pricing efficiency, which should come from the creation and redemption process, does not fully hold as equity ETFs show consistent price premiums. The tracking error of the funds is generally small and is decreasing over time. The risk of the ETF, daily price volatility and the total expense ratio explain a large part of the tracking error. Trading volume, fund size, bid-ask spread and average price premium or discount did not have an impact on the tracking error. Finally, it is concluded that market volatility and the tracking error are positively correlated.

Environmental isolation, biochemical identification, and antifungal drug susceptibility of Cryptococcus species

Introduction The incidence of opportunistic fungal infections has increased in recent years and is considered an important public health problem. Among systemic and opportunistic mycoses, cryptococcosis is distinguished by its clinical importance due to the increased risk of infection in individuals infected by human immunodeficiency virus. Methods To determine the occurrence of pathogenic Cryptococcus in pigeon excrement in the City of Araraquara, samples were collected from nine environments, including state and municipal schools, abandoned buildings, parks, and a hospital. The isolates were identified using classical tests, and susceptibility testing for the antifungal drugs (fluconazole, itraconazole, voriconazole, and amphotericin B) independently was also performed. After collection, the excrement samples were plated on Niger agar and incubated at room temperature. Results A total of 87 bird dropping samples were collected, and 66.6% were positive for the genus Cryptococcus. The following species were identified: Cryptococcus neoformans (17.2%), Cryptococcus gattii (5.2%), Cryptococcus ater (3.5%), Cryptococcus laurentti (1.7%), and Cryptococcus luteolus (1.7%). A total of 70.7% of the isolates were not identified to the species level and are referred to as Cryptococcus spp. throughout the manuscript. Conclusions Although none of the isolates demonstrated resistance to antifungal drugs, the identification of infested areas, the proper control of birds, and the disinfection of these environments are essential for the epidemiological control of cryptococcosis.

Evolutionary dynamics of Chlamydia trachomatis genome and identification of molecular patterns of hypothetical protein coding genes

The obligate intracellular bacterium Chlamydia trachomatis is a human pathogen of major public health significance. Strains can be classified into 15 main serovars (A to L3) that preferentially cause ocular infections (A-C), genital infections (D-K) or lymphogranuloma venereum (LGV) (L1-L3), but the molecular basis behind their distinct tropism, ecological success and pathogenicity is not welldefined. Most chlamydial research demands culture in eukaryotic cell lines, but it is not known if stains become laboratory adapted. By essentially using genomics and transcriptomics, we aimed to investigate the evolutionary patterns underlying the adaptation of C. trachomatis to the different human tissues, given emphasis to the identification of molecular patterns of genes encoding hypothetical proteins, and to understand the adaptive process behind the C. trachomatis in vivo to in vitro transition. Our results highlight a positive selection-driven evolution of C. trachomatis towards nichespecific adaptation, essentially targeting host-interacting proteins, namely effectors and inclusion membrane proteins, where some of them also displayed niche-specific expression patterns. We also identified potential "ocular-specific" pseudogenes, and pointed out the major gene targets of adaptive mutations associated with LGV infections. We further observed that the in vivo-derived genetic makeup of C. trachomatis is not significantly compromised by its long-term laboratory propagation. In opposition, its introduction in vitro has the potential to affect the phenotype, likely yielding virulence attenuation. In fact, we observed a "genital-specific" rampant inactivation of the virulence gene CT135, which may impact the interpretation of data derived from studies requiring culture. Globally, the findings presented in this Ph.D. thesis contribute for the understanding of C.trachomatis adaptive evolution and provides new insights into the biological role of C. trachomatishypothetical proteins. They also launch research questions for future functional studies aiming toclarify the determinants of tissue tropism, virulence or pathogenic dissimilarities among C. trachomatisstrains.

The efficacy of the old way/new way methodology on the correction of an automated technical error and its impact on the athlete’s psychological skills : Case study in tennis

The aim of this study was to evaluated the efficacy of the Old Way/New Way methodology (Lyndon, 1989/2000) with regard to the permanent correction of a consolidated and automated technical error experienced by a tennis athlete (who is 18 years old and has been engaged in practice mode for about 6 years) in the execution of serves. Additionally, the study assessed the impact of intervention on the athlete’s psychological skills. An individualized intervention was designed using strategies that aimed to produce a) a detailed analysis of the error using video images; b) an increased kinaesthetic awareness; c) a reactivation of memory error; d) the discrimination and generalization of the correct motor action. The athlete’s psychological skills were measured with a Portuguese version of the Psychological Skills Inventory for Sports (Cruz & Viana, 1993). After the intervention, the technical error was corrected with great efficacy and an increase in the athlete’s psychological skills was verified. This study demonstrates the methodology’s efficacy, which is consistent with the effects of this type of intervention in different contexts.

Gram-scale isolation of isobrucein B and neosergeolide from Picrolemma sprucei Hook. f.

Quassinoids neosergeolide and isobrucein B, obtained from Picrolemma sprucei, have proven in vitro antitumor, antimalarial, anthelminthic, cytotoxic, insecticide and leishmanicidal activities. There is interest in the in vivo pharmacological study of these natural compounds and their semi-synthetic derivatives, however, the quantities obtained in previous extraction processes have been shown to be a limiting factor for continuation of these studies. Herein, we describe a method for obtaining grams of these quassinoids whose purification relies only on recrystallization.

The isolation of Aspergillus spp from harvested maize in three Portuguese regions

In Portugal, maize is the cereal that involves more agriculture explorations. Aspergillus spp., among other species, are usually associated with this cereal, during drying and storage, making this commodity susceptible to mycotoxins (such as aflatoxins, ochratoxins, and cyclopiazonic acid). The aim of this study was to evaluate the mycotoxigenic potential of isolated Aspergillus strains from these maize samples and correlate it with the sampling place, the weather conditions, and local practices during drying and storage. The samples were collected between November 2008 and April 2009 in maize association of producers facilities in Coimbra, Santarém and Portalegre. The isolated strains were divided in three distinct groups, Aspergillus section Flavi, Aspergillus section Nigri and others Aspergillus. The preliminary results show that there are differences between the incidence of these groups in the three sampling places, especially in Coimbra, probably due to a lower mean temperatures and higher humidity levels. These data will be presented and discussed.

Modelo integrado de cuantificación del nivel de significación y de asignación del error tolerable en procesos de auditoría de estados contables

El objetivo que persigue un proceso de auditoría de estados contables es la comunicación por parte del auditor de una conclusión en relación al grado de razonabilidad con que tales estados reflejan la situación patrimonial, económica y financiera del ente de acuerdo a los criterios plasmados en las normas contables de referencia a ser utilizadas. El hecho que un auditor emita una conclusión errónea como consecuencia de su labor puede implicar la asunción de responsabilidades profesionales, civiles y penales como consecuencia de reclamos de usuarios de los estados contables que pudieran haberse visto perjudicados como consecuencia de la emisión de la conclusión errónea. Las normas contables a nivel nacional e internacional admiten la existencia de errores u omisiones en la información contenida en los estados contables, en la medida que tales desvíos no provoquen en los usuarios interesados en tales estados una decisión distinta a la que tomarían en caso de no existir los errores u omisiones aludidos. De lo expuesto en el párrafo anterior surge la cabal importancia que la determinación del nivel de significación total (nivel de desvíos admitidos por los usuarios de los estados contables en la información por ellos contenida) adquiere en los procesos de auditoría, como así también la asignación de tal nivel entre los distintos componentes de los estados contables (asignación del error tolerable) a los efectos de que los auditores eviten asumir responsabilidades de índole profesional, civil y/o penal. Hasta el momento no se conoce la existencia de modelos matemáticos que respalden de modo objetivo y verificable el cálculo del nivel de significación total y la asignación del error tolerable entre los distintos elementos conformantes de los estados contables. Entendemos que el desarrollo e integración de un modelo de cuantificación del nivel de significación total y de asignación del error tolerable tiene las siguientes repercusiones: 1 – Representaría para el auditor un elemento que respalde el modo de cuantificación del nivel de significación y la asignación del error tolerable entre los componentes de los estados contables. 2 – Permitiría que los auditores reduzcan las posibilidades de asumir responsabilidades de carácter profesional, civil y/o penales como consecuencia de su labor. 3 – Representaría un principio de avance a los efectos de que los organismos emisores de normas de auditoría a nivel nacional e internacional recepten elementos a los efectos de fijar directrices en relación al cálculo del nivel de significación y de asignación del error tolerable. 4 - Eliminaría al cálculo del nivel de significación como una barrera que afecte la comparabilidad de los estados contables.

Impacto de las prácticas agrícolas en los recursos hídricos de la provincia de Córdoba: aislamiento y caracterización de bacterias degradadoras de pesticidas para su utilización en procesos de biorremediación. Impact of agricultural activities on aquatic environments of Córdoba: isolation and characterization of bacteria with pesticide removal capacities for bioremediation treatments.

Los sistemas de agua dulce constituyen fuentes vitales para el desarrollo de la vida. Entre otras causas, el excesivo y poco controlado uso de pesticidas por las prácticas agrícolas modernas ha contribuido con la degradación de estos ecosistemas en la provincia de Córdoba (Secretaría de Ambiente, 2008). Los pesticidas son compuestos tóxicos y químicamente estables en el ambiente. Diversas especies microbianas presentan la capacidad para mineralizar dichos compuestos, siendo uno de los mecanismos de descomposición más importante. En el presente trabajo se pretende (1) Detectar y cuantificar principios activos de pesticidas en diferentes aguas ambientales de la región centro-sur de la provincia de Córdoba; (2) Aislar y caracterizar especies bacterianas a partir de muestras de aguas superficiales y subterráneas de la región que demuestren ser eficientes en la biodegradación de diferentes pesticidas. Para ello, se estudiará la presencia de 10 pesticidas organoclorados y organofosforados (lindano, 2,4D, DDT, p,p-DDE, �-endosulfán, �-endosulfán, clorpirifós, dimetoato) y herbicidas (atrazina) en muestras de agua superficiales y subterráneas (8 - 12m de profundidad) de la región agrícola centro-sur de Córdoba. Se establecerán diferentes puntos de muestreo en las principales cuencas hidrográficas: Río Tercero y Embalse de Río Tercero, Canal Desviador de Bell Ville, Laguna La Salada, Río Saladillo, Río Carcarañá y Río Cuarto. La determinación de pesticidas se realizará mediante Cromatografía de Gases (GC). Conjuntamente, se realizará el aislamiento de microorganismos a partir de las muestras de agua suplementadas con diferentes concentraciones de pesticidas (López et al., 2005) y se procederá a la caracterización morfológica y bioquímica (Lechevalier, 1989). Se realizarán curvas de crecimiento (DO600) y se determinará la viabilidad celular mediante el método de recuento en placa. El potencial catabólico de cada aislamiento se determinará analizando la concentración residual de pesticidas en el sobrenadante de los cultivos (Benimeli et al, 2003) y mediante ensayos de resting-cell (Hernández et al, 2008). Finalmente se realizará la caracterización genética (Weisburg, 1991) de los aislamientos que demuestren una mayor eficiencia en la biodegradación de pesticidas. El presente estudio pretende abordar una temática prioritaria como es la contaminación ambiental de ecosistemas acuáticos de la provincia de Córdoba. la detección de principios activos de pesticidas sería, por lo tanto, un indicador de contaminación de origen antropológica y brindaría información respecto al deterioro de la calidad del agua. Por otra parte, el aislamiento de bacterias adaptadas a las condiciones ecológicas de la región y capaces de metabolizar eficientemente diferentes pesticidas como fuentes de carbono y energía sería beneficioso para su utilización en futuros ensayos de biorremediación.

Non-classical error bounds in the central limit theorem

The classical central limit theorem states the uniform convergence of the distribution functions of the standardized sums of independent and identically distributed square integrable real-valued random variables to the standard normal distribution function. While first versions of the central limit theorem are already due to Moivre (1730) and Laplace (1812), a systematic study of this topic started at the beginning of the last century with the fundamental work of Lyapunov (1900, 1901). Meanwhile, extensions of the central limit theorem are available for a multitude of settings. This includes, e.g., Banach space valued random variables as well as substantial relaxations of the assumptions of independence and identical distributions. Furthermore, explicit error bounds are established and asymptotic expansions are employed to obtain better approximations. Classical error estimates like the famous bound of Berry and Esseen are stated in terms of absolute moments of the random summands and therefore do not reflect a potential closeness of the distributions of the single random summands to a normal distribution. Non-classical approaches take this issue into account by providing error estimates based on, e.g., pseudomoments. The latter field of investigation was initiated by work of Zolotarev in the 1960's and is still in its infancy compared to the development of the classical theory. For example, non-classical error bounds for asymptotic expansions seem not to be available up to now ...