New insights into host-parasite ubiquitin proteome dynamics in P. falciparum infected red blood cells using a TUBEs-MS approach

Malaria, caused by Plasmodium falciparum (P. falciparum), ranks as one of the most baleful infectious diseases worldwide. New antimalarial treatments are needed to face existing or emerging drug resistant strains. Protein degradation appears to play a significant role during the asexual intraerythrocytic developmental cycle (IDC) of P. falciparum. Inhibition of the ubiquitin proteasome system (UPS), a major intracellular proteolytic pathway, effectively reduces infection and parasite replication. P. falciparum and erythrocyte UPS coexist during IDC but the nature of their relationship is largely unknown. We used an approach based on Tandem Ubiquitin-Binding Entities (TUBEs) and 1D gel electrophoresis followed by mass spectrometry to identify major components of the TUBEs-associated ubiquitin proteome of both host and parasite during ring, trophozoite and schizont stages. Ring-exported protein (REX1), a P. falciparum protein located in Maurer's clefts and important for parasite nutrient import, was found to reach a maximum level of ubiquitylation in trophozoites stage. The Homo sapiens (H. sapiens) TUBEs associated ubiquitin proteome decreased during the infection, whereas the equivalent P. falciparum TUBEs-associated ubiquitin proteome counterpart increased. Major cellular processes such as DNA repair, replication, stress response, vesicular transport and catabolic events appear to be regulated by ubiquitylation along the IDC P. falciparum infection.

New insights into functional regulation in MS-based drug profiling

We present a novel data analysis strategy which combined with subcellular fractionation and liquid chromatography-mass spectrometry (LC-MS) based proteomics provides a simple and effective workflow for global drug profiling. Five subcellular fractions were obtained by differential centrifugation followed by high resolution LC-MS and complete functional regulation analysis. The methodology combines functional regulation and enrichment analysis into a single visual summary. The workflow enables improved insight into perturbations caused by drugs. We provide a statistical argument to demonstrate that even crude subcellular fractions leads to improved functional characterization. We demonstrate this data analysis strategy on data obtained in a MS-based global drug profiling study. However, this strategy can also be performed on other types of large scale biological data.

Design of a sea-level tsunami detection network for the Gulf of Cadiz

The devastating impact of the Sumatra tsunami of 26 December 2004, raised the question for scientists of how to forecast a tsunami threat. In 2005, the IOC-UNESCO XXIII assembly decided to implement a global tsunami warning system to cover the regions that were not yet protected, namely the Indian Ocean, the Caribbean and the North East Atlantic, the Mediterranean and connected seas (the NEAM region). Within NEAM, the Gulf of Cadiz is the more sensitive area, with an important record of devastating historical events. The objective of this paper is to present a preliminary design for a reliable tsunami detection network for the Gulf of Cadiz, based on a network of sea-level observatories. The tsunamigenic potential of this region has been revised in order to define the active tectonic structures. Tsunami hydrodynamic modeling and GIS technology have been used to identify the appropriate locations for the minimum number of sea-level stations. Results show that 3 tsunameters are required as the minimum number of stations necessary to assure an acceptable protection to the large coastal population in the Gulf of Cadiz. In addition, 29 tide gauge stations could be necessary to fully assess the effects of a tsunami along the affected coasts of Portugal, Spain and Morocco.

Epidemiological methods for research with drug misusers: review of methods for studying prevalence and morbidity

Epidemiological studies of drug misusers have until recently relied on two main forms of sampling: probability and convenience. The former has been used when the aim was simply to estimate the prevalence of the condition and the latter when in depth studies of the characteristics, profiles and behaviour of drug users were required, but each method has its limitations. Probability samples become impracticable when the prevalence of the condition is very low, less than 0.5% for example, or when the condition being studied is a clandestine activity such as illicit drug use. When stratified random samples are used, it may be difficult to obtain a truly representative sample, depending on the quality of the information used to develop the stratification strategy. The main limitation of studies using convenience samples is that the results cannot be generalised to the whole population of drug users due to selection bias and a lack of information concerning the sampling frame. New methods have been developed which aim to overcome some of these difficulties, for example, social network analysis, snowball sampling, capture-recapture techniques, privileged access interviewer method and contact tracing. All these methods have been applied to the study of drug misuse. The various methods are described and examples of their use given, drawn from both the Brazilian and international drug misuse literature.

Hippuric acid in urine: reference values

OBJECTIVE: To establish reference values for hippuric acid (HA) excreted in the urine, and to evaluate the impact of age, gender, alcohol, and tobacco, on these levels in a population nonexposed to toluene. METHODS: Reference values for hippuric acid in urine were determined in 115 toluene nonexposed healthy volunteers, from Alfenas city, Southeastern Brazil. A questionnaire was applied to each volunteer and data on occupational and personal habits were collected. Biochemical and hematological analyses were used to confirm the volunteers' good health condition. Reference values were expressed in g HA/g urine creatinine, as mean ± standard deviation (x ± SD), median, 95% confidence interval (95%CI), 95th percentile, and upper reference value (URV, mean +2 SD). RESULTS: Reference values of hippuric acid in urine were: mean ± standard deviation =0.18±0.10; median =0.15; 95% confidence interval =0.16±0.20; 95th percentile = 0.36 and upper reference value (URV, mean +2 SD) =0.38. Statistically significant differences in urinary HA (Wilcoxon - Mann/Whitney, p<0.05) were observed for different genders and age groups. Alcohol ingestion and smoking habit did not significantly affect the results. CONCLUSIONS: The reference values of hippuric acid in urine can be used in biomonitoring programs of workers occupationally exposed to toluene, especially in the southern region of the state of Minas Gerais. Age and gender may affect the HA reference values.

Risk of drug interaction: combination of antidepressants and other drugs

OBJECTIVE: To assess the frequency of combination of antidepressants with other drugs and risk of drug interactions in the setting public hospital units in Brazil. METHODS: Prescriptions of all patients admitted to a public hospital from November 1996 to February 1997 were surveyed from the hospital's data processing center in São Paulo, Brazil. A manual search of case notes of all patients admitted to the psychiatric unit from January 1993 to December 1995 and all patients registered in the affective disorders outpatient clinic in December 1996 was carried out. Patients taking any antidepressant were identified and concomitant use of drugs was checked. By means of a software program (Micromedex®) drug interactions were identified. RESULTS: Out of 6,844 patients admitted to the hospital, 63 (0.9%) used antidepressants and 16 (25.3%) were at risk of drug interaction. Out of 311 patients in the psychiatric unit, 63 (20.2%) used antidepressants and 13 of them (20.6%) were at risk. Out of 87 patients in the affective disorders outpatient clinic, 43 (49.4%) took antidepressants and 7 (16.2%) were at risk. In general, the use of antidepressants was recorded in 169 patients and 36 (21.3%) were at risk of drug interactions. Twenty different forms of combinations at risk of drug interactions were identified: four were classified as mild, 15 moderate and one severe interaction. CONCLUSION: In the hospital general units the number of drug interactions per patient was higher than in the psychiatric unit; and prescription for depression was lower than expected.

VPP´s multi-level negotiation in smart grids and competitive electricity markets

The increase of distributed generation (DG) has brought about new challenges in electrical networks electricity markets and in DG units operation and management. Several approaches are being developed to manage the emerging potential of DG, such as Virtual Power Players (VPPs), which aggregate DG plants; and Smart Grids, an approach that views generation and associated loads as a subsystem. This paper presents a multi-level negotiation mechanism for Smart Grids optimal operation and negotiation in the electricity markets, considering the advantages of VPPs’ management. The proposed methodology is implemented and tested in MASCEM – a multiagent electricity market simulator, developed to allow deep studies of the interactions between the players that take part in the electricity market negotiations.

Drug therapy complexity: the importance of pharmacotherapeutic follow-up

Nowadays, the phenomenon of population ageing represents an worldwide problem, which assumes particular significance in Portugal. As they get older, individuals present more comorbidities and consequently consume an increasing number of drugs, which contributes to a growing drug therapy complexity. The institutionalized elders are particularly affected by this occurrence. Drug therapy complexity is defined as the conciliator of several characteristics of the pharmacotherapy and can affect patient’s safety and medication adherence. It can be measured with Medication Regimen Complexity Index (MRCI). This study aims to determine the drug therapy complexity of institutionalized elders in order to assess the need of pharmacotherapeutic follow-up.

Chemical and biological studies from an Azorean macroalga: Ulva rigida

3rd Portuguese Meeting on Medicinal Chemistry and 1st Portuguese-Spanish-Brazilian Meeting on Medicinal Chemistry, Aveiro, 28-30 Novembro 2012.

Determinants of salivary cotinine level: a population-based study in Brazil

OBJECTIVE: A cross-sectional population-based study was conducted to assess, in active smokers, the relationship of number of cigarettes smoked and other characteristics to salivary cotinine concentrations. METHODS: A random sample of active smokers aged 15 years or older was selected using a stepwise cluster sample strategy, in the year 2000 in Rio de Janeiro, Brazil. The study included 401 subjects. Salivary cotinine concentration was determined using gas chromatography with nitrogen-phosphorus detection. A standard questionnaire was used to collect demographic and smoking behavioral data. The relation between the number of cigarettes smoked in the last 24h and cotinine level was examined by means of a nonparametric fitting technique of robust locally weighted regression. RESULTS: Significantly (p<0.05) higher adjusted mean cotinine levels were found in subjects smoking their first cigarette within five minutes after waking up, and in those smoking 1-20 cigarettes in the last 24h who reported inhaling more than ½ the time. In those smoking 1-20 cigarettes, the slope was significantly higher for those subjects waiting for more than five minutes before smoking their first cigarette after waking up, and those smoking "light" cigarettes when compared with their counterparts. These heterogeneities became negligible and non-significant when subjects with cotinine >40 ng/mL per cigarette were excluded. CONCLUSIONS: There was found a positive association between self-reporting smoking five minutes after waking up, and inhaling more than ½ the time are consistent and higher cotinine levels. These can be markers of dependence and higher nicotine intake. Salivary cotinine proved to be a useful biomarker of recent smoking and can be used in epidemiological studies and smoking cessation programs.

Core group approach to identify college students at risk for sexually transmitted infections

OBJECTIVE: To analyze the core group for sexually transmitted infections (STI) among college students. METHODS: Cross-sectional study carried out in a convenience sample comprising 711 college students of the public university of Morelos, Mexico, between 2001 and 2003. Sociodemographic and sexual behavior information were collected using self-applied questionnaires. Herpes simplex 2 (HSV-2) infection was tested in the blood. The number of sexual partners in the last year and cocaine consumption were used as indicators to construct the dependent variable "level of STI risk" in three categories: low, medium and high risk (core group). A multinomial analysis was conducted to evaluate whether different sex behaviors were associated with the variable "level of STI risk". RESULTS: There was significant association between HSV-2 seroprevalence and the variable "level of STI risk": 13%, 5.6% and 3.8% were found in high (core group), medium and low categories, respectively. There were gender differences regarding the core group. Men started having sexual intercourse earlier, had more sex partners, higher alcohol and drug consumption, higher frequency of sex intercourse with sex workers, exchanging sex for money, occasional and concurrent partners compared to women. CONCLUSIONS: The study findings suggest existing contextual characteristics in the study population that affect their sex behavior. In Mexico, the cultural conception of sexuality is determined mainly by gender differences where men engage in higher risky sexual behavior than women.

Homicide and impunity: an ecological analysis at state level in Brazil

OBJECTIVE: To assess a new impunity index and variables that have been found to predict variation in homicide rates in other geographical levels as predictive of state-level homicide rates in Brazil. METHODS: This was a cross-sectional ecological study. Data from the mortality information system relating to the 27 Brazilian states for the years 1996 to 2005 were analyzed. The outcome variables were taken to be homicide victim rates in 2005, for the entire population and for men aged 20-29 years. Measurements of economic and social development, economic inequality, demographic structure and life expectancy were analyzed as predictors. An "impunity index", calculated as the total number of homicides between 1996 and 2005 divided by the number of individuals in prison in 2007, was constructed. The data were analyzed by means of simple linear regression and negative binomial regression. RESULTS: In 2005, state-level crude total homicide rates ranged from 11 to 51 per 100,000; for young men, they ranged from 39 to 241. The impunity index ranged from 0.4 to 3.5 and was the most important predictor of this variability. From negative binomial regression, it was estimated that the homicide victim rate among young males increased by 50% for every increase of one point in this ratio. CONCLUSIONS: Classic predictive factors were not associated with homicides in this analysis of state-level variation in Brazil. However, the impunity index indicated that the greater the impunity, the higher the homicide rate.

Stability of the tree-level vacuum in two Higgs doublet models against charge or CP spontaneous violation

We show that in two Higgs doublet models at tree-level the potential minimum preserving electric charge and CP symmetries, when it exists, is the global one. Furthermore, we derived a very simple condition, involving only the coefficients of the quartic terms of the potential, that guarantees spontaneous CP breaking. (C) 2004 Elsevier B.V. All rights reserved.

Sustainable blended-learning in HEI: developing and implementing multi-level interventions

This chapter aims to demonstrate how PAOL - Unit for Innovation in Education, a project from ISCAP - School of Accounting and Administration of Oporto ....