Computational analysis and physico-chemical characterization of an inclusion compound between praziquantel and methyl-beta-cyclodextrin for use as an alternative in the treatment of schistosomiasis

Schistosomiasis is still an endemic disease in many regions, with 250 million people infected with Schistosoma and about 500,000 deaths per year. Praziquantel (PZQ) is the drug of choice for schistosomiasis treatment, however it is classified as Class II in the Biopharmaceutics Classification System, as its low solubility hinders its performance in biological systems. The use of cyclodextrins is a useful tool to increase the solubility and bioavailability of drugs. The aim of this work was to prepare an inclusion compound of PZQ and methyl-beta-cyclodextrin (MeCD), perform its physico-chemical characterization, and explore its in vitro cytotoxicity. SEM showed a change of the morphological characteristics of PZQ:MeCD crystals, and IR data supported this finding, with changes after interaction with MeCD including effects on the C-H of the aromatic ring, observed at 758 cm(-1). Differential scanning calorimetry measurements revealed that complexation occurred in a 1:1 molar ratio, as evidenced by the lack of a PZQ transition temperature after inclusion into the MeCD cavity. In solution, the PZQ UV spectrum profile in the presence of MeCD was comparable to the PZQ spectrum in a hydrophobic solvent. Phase solubility diagrams showed that there was a 5.5-fold increase in PZQ solubility, and were indicative of a type A(L) isotherm, that was used to determine an association constant (K(a)) of 140.8 M(-1). No cytotoxicity of the PZQ:MeCD inclusion compound was observed in tests using 3T3 cells. The results suggest that the association of PZQ with MeCD could be a good alternative for the treatment of schistosomiasis.

A self-inducible heterologous protein expression system in Escherichia coli

International audience

Early Warning System Potential for Single Sourcing Risk Mitigation

Part 17: Risk Analysis

EXISTENCE OF UNUSED MANAGED CAPACITY ON DEDICATED LANES AND AN ALTERNATIVE ON HOW TO SELL IT VIA AUCTIONS

This dissertation verifies whether the following two hypotheses are true: (1) High-occupancy/toll lanes (and therefore other dedicated lanes) have capacity that could still be used; (2) such unused capacity (or more precisely, “unused managed capacity”) can be sold successfully through a real-time auction. To show that the second statement is true, this dissertation proposes an auction-based metering (ABM) system, that is, a mechanism that regulates traffic that enters the dedicated lanes. Participation in the auction is voluntary and can be skipped by paying the toll or by not registering to the new system. This dissertation comprises the following four components: a measurement of unused managed capacity on an existing HOT facility, a game-theoretic model of an ABM system, an operational description of the ABM system, and a simulation-based evaluation of the system. Some other and more specific contributions of this dissertation include the following: (1) It provides a definition and a methodology for measuring unused managed capacity and another important variable referred as “potential volume increase”. (2) It proves that the game-theoretic model has a unique Bayesian Nash equilibrium. (3) And it provides a specific road design that can be applied or extended to other facilities. The results provide evidence that the hypotheses are true and suggest that the ABM system would benefit a public operator interested in reducing traffic congestion significantly, would benefit drivers when making low-reliability trips (such as work-to-home trips), and would potentially benefit a private operator interested in raising revenue.

A regulation-based classification system for marine protected areas (MPAs)

Dados suplementares associados com este artigo disponíveis na versão online em: http://dx.doi.org/10.1016/j.marpol.2016.06.021

RISKS TO FOOD AVAILABILITY AND ACCESS FROM CLIMATE POLICIES: AN INTEGRATED ASSESSMENT OF REGIONAL FOOD AVAILABILITY AND ACCESS WITH ALTERNATIVE CLIMATE MITIGATION STRATEGIES TO 2050

Although mitigating GHG emissions is necessary to reduce the overall negative climate change impacts on crop yields and agricultural production, certain mitigation measures may generate unintended consequences to food availability and access due to land use competition and economic burden of mitigation. Prior studies have examined the co-impacts on food availability and global producer prices caused by alternative climate policies. More recent studies have looked at the reduction in total caloric intake driven by both changing income and changing food prices under one specific climate policy. However, due to inelastic calorie demand, consumers’ well-being are likely further reduced by increased food expenditures. Built upon existing literature, my dissertation explores how alternative climate policy designs might adversely affect both caloric intake and staple food budget share to 2050, by using the Global Change Assessment Model (GCAM) and a post-estimated metric of food availability and access (FAA). My dissertation first develop a set of new metrics and methods to explore new perspectives of food availability and access under new conditions. The FAA metric consists of two components, the fraction of GDP per capita spent on five categories of staple food and total caloric intake relative to a reference level. By testing the metric against alternate expectations of the future, it shows consistent results with previous studies that economic growth dominates the improvement of FAA. As we increase our ambition to achieve stringent climate targets, two policy conditions tend to have large impacts on FAA driven by competing land use and increasing food prices. Strict conservation policies leave the competition between bioenergy and agriculture production on existing commercial land, while pricing terrestrial carbon encourages large-scale afforestation. To avoid unintended outcomes to food availability and access for the poor, pricing land emissions in frontier forests has the advantage of selecting more productive land for agricultural activities compared to the full conservation approach, but the land carbon price should not be linked to the price of energy system emissions. These results are highly relevant to effective policy-making to reduce land use change emissions, such as the Reduced Emissions from Deforestation and Forest Degradation (REDD).

Quantifying the Resilience of an Urban Traffic-Electric Power Coupled System

Transportation system resilience has been the subject of several recent studies. To assess the resilience of a transportation network, however, it is essential to model its interactions with and reliance on other lifelines. In this work, a bi-level, mixed-integer, stochastic program is presented for quantifying the resilience of a coupled traffic-power network under a host of potential natural or anthropogenic hazard-impact scenarios. A two-layer network representation is employed that includes details of both systems. Interdependencies between the urban traffic and electric power distribution systems are captured through linking variables and logical constraints. The modeling approach was applied on a case study developed on a portion of the signalized traffic-power distribution system in southern Minneapolis. The results of the case study show the importance of explicitly considering interdependencies between critical infrastructures in transportation resilience estimation. The results also provide insights on lifeline performance from an alternative power perspective.

Targeted gene modification in Fragaria vesca mediated by CRISPR/Cas9 system

Genome editing is becoming an important biotechnological tool for gene function analysis and crop improvement, being the CRISPR-Cas9 (Clustered Regularly Interspaced Short Palindromic Repeat-CRISPR associated protein 9) system the most widely used. The natural CRISPR/Cas9 system has been reduced to two components: a single-guide RNA (sgRNA) for target recognition via RNA-DNA base pairing, which is commonly expressed using a promoter for small-RNAs (U6 promoter), and the Cas9 endonuclease for DNA cleavage (1). To validate the CRISPR/Cas9 system in strawberry plants, we designed two sgRNAs directed against the floral homeotic gene APETALA3 (sgRNA-AP3#1 and sgRNA-AP3#2). This gene was selected because ap3 mutations induce clear developmental phenotypes in which petals and stamens are missing or partially converted to sepals and carpels respectively (2). In this work, we used two different U6 promoters to drive the sgRNA-AP3s expression: AtU6-26 from Arabidopsis (4), and a U6 promoter from Fragaria vesca (FvU6) (this work). We also tested two different coding sequences of Cas9: a human- (hSpCas9) (3) and a plant-codon optimized (pSpCas9) (this work). Transient expression experiments using both CRISPR/Cas9 systems (AtU6-26:sgRNA-AP3#1_35S:hSpCas9_AtU6-26:sgRNA-AP3#2 and FvU6:sgRNA-AP3#1_35S:pSpCas9_FvU6:sgRNA-AP3#2) were performed infiltrating Agrobacterium tumefaciens into F. vesca fruits. PCR amplification and sequencing analyses across the target sites showed a deletion of 188-189 bp corresponding to the region comprised between the two cutting sites of Cas9, confirming that the CRISPR/Cas9 system is functional in F. vesca. Remarkably, the two systems showed different mutagenic efficiency that could be related to differences in expression of the U6 promoters as well as differences in the Cas9 transcripts stability and translation. Stable transformants for both F. vesca (2n) and Fragaria X anannassa (8n) are currently being established to test whether is possible to obtain heritable homozygous mutants derived from CRISPR/Cas9 strategies in strawberry. Thus, our work offers a promising tool for genome editing and gene functional analysis in strawberry. This tool might represent a more efficient alternative to the sometimes inefficient RNAi silencing methods commonly used in this species.

Hemolytic activity of alternative complement pathway as an indicator of innate immunity in pacu (Piaractus mesopotamicus)

The objective of this study was to evaluate the methodology to establish the hemolytic activity of alternative complement pathway as an indicator of the innate immunity in Brazilian fish pacu (Piaractus mesopotamicus), in addition to verifying the influence of β-glucan as an immunostimulant. Fish were fed with diets containing 0, 0.1 and 1% β-glucan, during seven days, and then inoculated with Aeromonas hydrophila. Seven days after the challenge, they were bled for serum extraction. The methodology consisted of a kinetic assay that allows calculating the required time for serum proteins of the complement to promote 50% lysis of a rabbit red blood cell suspension. The method developed in mammals was successfully applied for pacu and determined that the hemolytic activity of the proteins of the complement system (alternative pathway) increased after the pathogen challenge, but was not influenced by the β-glucan treatment.

The Planned and the Emergent: An Alternative Model of Learning and Literacy

Within academic institutions, writing centers are uniquely situated, socially rich sites for exploring learning and literacy. I examine the work of the Michigan Tech Writing Center's UN 1002 World Cultures study teams primarily because student participants and Writing Center coaches are actively engaged in structuring their own learning and meaning-making processes. My research reveals that learning is closely linked to identity formation and leading the teams is an important component of the coaches' educational experiences. I argue that supporting this type of learning requires an expanded understanding of literacy and significant changes to how learning environments are conceptualized and developed. This ethnographic study draws on data collected from recordings and observations of one semester of team sessions, my own experiences as a team coach and UN 1002 teaching assistant, and interviews with Center coaches prior to their graduation. I argue that traditional forms of assessment and analysis emerging from individualized instruction models of learning cannot fully account for the dense configurations of social interactions identified in the Center's program. Instead, I view the Center as an open system and employ social theories of learning and literacy to uncover how the negotiation of meaning in one context influences and is influenced by structures and interactions within as well as beyond its boundaries. I focus on the program design, its enaction in practice, and how engagement in this type of writing center work influences coaches' learning trajectories. I conclude that, viewed as participation in a community of practice, the learning theory informing the program design supports identity formation —a key aspect of learning as argued by Etienne Wenger (1998). The findings of this study challenge misconceptions of peer learning both in writing centers and higher education that relegate peer tutoring to the role of support for individualized models of learning. Instead, this dissertation calls for consideration of new designs that incorporate peer learning as an integral component. Designing learning contexts that cultivate and support the formation of new identities is complex, involves a flexible and opportunistic design structure, and requires the availability of multiple forms of participation and connections across contexts.

Feasibility study and flow scheduling optimization of a multi-product pipeline system

The first goal of this study is to analyse a real-world multiproduct onshore pipeline system in order to verify its hydraulic configuration and operational feasibility by constructing a simulation model step by step from its elementary building blocks that permits to copy the operation of the real system as precisely as possible. The second goal is to develop this simulation model into a user-friendly tool that one could use to find an “optimal” or “best” product batch schedule for a one year time period. Such a batch schedule could change dynamically as perturbations occur during operation that influence the behaviour of the entire system. The result of the simulation, the ‘best’ batch schedule is the one that minimizes the operational costs in the system. The costs involved in the simulation are inventory costs, interface costs, pumping costs, and penalty costs assigned to any unforeseen situations. The key factor to determine the performance of the simulation model is the way time is represented. In our model an event based discrete time representation is selected as most appropriate for our purposes. This means that the time horizon is divided into intervals of unequal lengths based on events that change the state of the system. These events are the arrival/departure of the tanker ships, the openings and closures of loading/unloading valves of storage tanks at both terminals, and the arrivals/departures of trains/trucks at the Delivery Terminal. In the feasibility study we analyse the system’s operational performance with different Head Terminal storage capacity configurations. For these alternative configurations we evaluated the effect of different tanker ship delay magnitudes on the number of critical events and product interfaces generated, on the duration of pipeline stoppages, the satisfaction of the product demand and on the operative costs. Based on the results and the bottlenecks identified, we propose modifications in the original setup.

Reforming the Swedish employment-related social security system:activation, administrative modernization and strengthening local autonomy

The Swedish system of social security has often been regarded as comprehensive and comprehensive and inclusive. During major reforms in the 1990s and 2000s, it has maintained its essential character as a popular and well-endowed provider of social security and stability. Employment-related benefits are generous in financial terms, but come with the need for recipients to remain actively engaged in the economic or educational field. However, Sweden’s geographical and demographic diversity made it necessary to increase the role of local authorities in implementing active labour market policies. This article tracks these developments since the mid-1990s, both with regard to changing the benefits system and with regard to changing local government involvement. It argues that backed by broad political support, the Swedish system has achieved the necessary modernisation and adaptation to remain a viable alternative to more neo-liberal welfare retrenchment projects conducted in other European countries.

Assessment of MALDI-TOF MS as Alternative Tool for Streptococcus suis Identification

The accuracy of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for identifying Streptococcus suis isolates obtained from pigs, wild animals, and humans was evaluated using a PCR-based identification assay as the gold standard. In addition, MALDI-TOF MS was compared with the commercial multi-tests Rapid ID 32 STREP system. From the 129 S. suis isolates included in the study and identified by the molecular method, only 31 isolates (24.03%) had score values ≥2.300 and 79 isolates (61.24%) gave score values between 2.299 and 2.000. After updating the currently available S. suis MALDI Biotyper database with the spectra of three additional clinical isolates of serotypes 2, 7, and 9, most isolates had statistically significant higher score values (mean score: 2.65) than those obtained using the original database (mean score: 2.182). Considering the results of the present study, we suggest using a less restrictive threshold score of ≥2.000 for reliable species identification of S. suis. According to this cut-off value, a total of 125 S. suis isolates (96.9%) were correctly identified using the updated database. These data indicate an excellent performance of MALDI-TOF MS for the identification of S. suis.

Development of a responsive fisheries management system for the Portuguese crustacean bottom trawl fishery: Lessons learnt

A prototype for a Responsive Fisheries Management System (RFMS) was developed in the context of the European FP7 project EcoFishMan and tested on the Portuguese crustacean trawl fishery. Building on Results Based Management principles, RFMS involves the definition of specific and measurable objectives for a fishery by the relevant authorities but allows resource users the freedom to find ways to achieve the objectives and to provide adequate documentation. Taking into account the main goals of the new Common Fisheries Policy, such as sustainable utilization of the resources, end of discards and unwanted catches, a management plan for the Portuguese crustacean trawl fishery was developed in cooperation with the fishing industry, following the process and design laid out in the RFMS concept. The plan considers biological, social and economic goals and assigns a responsibility for increased data collection to the resource users. The performance of the plan with regard to selected indicators was evaluated through simulations. In this paper the process towards a RFMS is described and the lessons learnt from the interaction with stakeholders in the development of an alternative management plan are discussed. (C) 2014 Elsevier Ltd. All rights reserved.

A regulation-based classification system for marine protected areas (MPAs)

Marine protected areas (MPAs) are a global conservation and management tool to enhance the resilience of linked social-ecological systems with the aim of conserving biodiversity and providing ecosystem services for sustainable use. However, MPAs implemented worldwide include a large variety of zoning and management schemes from single to multiple-zoning and from no-take to multiple-use areas. The current IUCN categorisation of MPAs is based on management objectives which many times have a significant mismatch to regulations causing a strong uncertainty when evaluating global MPAs effectiveness. A novel global classification system for MPAs based on regulations of uses as an alternative or complementing, the current IUCN system of categories is presented. Scores for uses weighted by their potential impact on biodiversity were built. Each zone within a MPA was scored and an MPA index integrates the zone scores. This system classifies MPAs as well as each MPA zone individually, is globally applicable and unambiguously discriminates the impacts of uses. (C) 2016 The Authors. Published by Elsevier Ltd.