A Practical Teaching Course in Directed Protein Evolution Using the Green Fluorescent Protein as a Model

Protein engineering is a powerful tool, which correlates protein structure with specific functions, both in applied biotechnology and in basic research. Here, we present a practical teaching course for engineering the green fluorescent protein (GFP) from Aequorea victoria by a random mutagenesis strategy using error-prone polymerase chain reaction. Screening of bacterial colonies transformed with random mutant libraries identified GFP variants with increased fluorescence yields. Mapping the three-dimensional structure of these mutants demonstrated how alterations in structural features such as the environment around the fluorophore and properties of the protein surface can influence functional properties such as the intensity of fluorescence and protein solubility.

Effect of a sequential education and monitoring programme on quality-of-life components in heart failure

Aims Trials of disease management programmes (DMP) in heart failure (HF) have shown controversial results regarding quality of life. We hypothesized that a DMP applied over the long-term could produce different effects on each of the quality-of-life components. Methods and results We extended the prospective, randomized REMADHE Trial, which studied a DMP in HF patients. We analysed changes in Minnesota Living with Heart Failure Questionnaire components in 412 patients, 60.5% male, age 50.2 +/- 11.4 years, left ventricular ejection fraction 34.7 +/- 10.5%. During a mean follow-up of 3.6 +/- 2.2 years, 6.3% of patients underwent heart transplantation and 31.8% died. Global quality-of-life scores improved in the DMP intervention group, compared with controls, respectively: 57.5 +/- 3.1 vs. 52.6 +/- 4.3 at baseline, 32.7 +/- 3.9 vs. 40.2 +/- 6.3 at 6 months, 31.9 +/- 4.3 vs. 41.5 +/- 7.4 at 12 months, 26.8 +/- 3.1 vs. 47.0 +/- 5.3 at the final assessment; P<0.01. Similarly, the physical component (23.7 +/- 1.4 vs. 21.1 +/- 2.2 at baseline, 16.2 +/- 2.9 vs. 18.0 +/- 3.3 at 6 months, 17.3 +/- 2.9 vs. 23.1 +/- 5.7 at 12 months, 11.4 +/- 1.6 vs. 19.9 +/- 2.4 final; P<0.01), the emotional component (13.2 +/- 1.0 vs. 12.1 +/- 1.4 at baseline, 11.7 +/- 2.7 vs. 12.3 +/- 3.1 at 6 months, 12.4 +/- 2.9 vs. 16.8 +/- 5.9 at 12 months, 6.7 +/- 1.0 vs. 10.6 +/- 1.4 final; P<0.01) and the additional questions (20.8 +/- 1.2 vs. 19.3 +/- 1.8 at baseline, 14.3 +/- 2.7 vs. 17.3 +/- 3.1 at 6 months, 12.4 +/- 2.9 vs. 21.0 +/- 5.5 at 12 months, 6.7 +/- 1.4 vs. 17.3 +/- 2.2 final; P<0.01) were better (lower) in the intervention group. The emotional component improved earlier than the others. Post-randomization quality of life was not associated with events. Conclusion Components of the quality-of-life assessment responded differently to DMP. These results indicate the need for individualized DMP strategies in patients with HF. Trial registration information www.clincaltrials.gov NCT00505050-REMADHE.

A practical approach to assess depression risk and to guide risk reduction strategies in later life

Background: Many factors have been associated with the onset and maintenance of depressive symptoms in later life, although this knowledge is yet to be translated into significant health gains for the population. This study gathered information about common modifiable and non-modifiable risk factors for depression with the aim of developing a practical probabilistic model of depression that can be used to guide risk reduction strategies. \Methods: A cross-sectional study was undertaken of 20,677 community-dwelling Australians aged 60 years or over in contact with their general practitioner during the preceding 12 months. Prevalent depression (minor or major) according to the Patient Health Questionnaire (PHQ-9) assessment was the main outcome of interest. Other measured exposures included self-reported age, gender, education, loss of mother or father before age 15 years, physical or sexual abuse before age 15 years, marital status, financial stress, social support, smoking and alcohol use, physical activity, obesity, diabetes, hypertension, and prevalent cardiovascular diseases, chronic respiratory diseases and cancer. Results: The mean age of participants was 71.7 +/- 7.6 years and 57.9% were women. Depression was present in 1665 (8.0%) of our subjects. Multivariate logistic regression showed depression was independently associated with age older than 75 years, childhood adverse experiences, adverse lifestyle practices (smoking, risk alcohol use, physical inactivity), intermediate health hazards (obesity, diabetes and hypertension), comorbid medical conditions (clinical history of coronary heart disease, stroke, asthma, chronic obstructive pulmonary disease, emphysema or cancers), and social or financial strain. We stratified the exposures to build a matrix that showed that the probability of depression increased progressively with the accumulation of risk factors, from less than 3% for those with no adverse factors to more than 80% for people reporting the maximum number of risk factors. Conclusions: Our probabilistic matrix can be used to estimate depression risk and to guide the introduction of risk reduction strategies. Future studies should now aim to clarify whether interventions designed to mitigate the impact of risk factors can change the prevalence and incidence of depression in later life.

Enhanced classification of Chagas serologic results and epidemiologic characteristics of seropositive donors at three large blood centers in Brazil

BACKGROUND: A major problem in Chagas disease donor screening is the high frequency of samples with inconclusive results. The objective of this study was to describe patterns of serologic results among donors to the three Brazilian REDS-II blood centers and correlate with epidemiologic characteristics. STUDY DESIGN AND METHODS: The centers screened donor samples with one Trypanosoma cruzi lysate enzyme immunoassay (EIA). EIA-reactive samples were tested with a second lysate EIA, a recombinant-antigen based EIA, and an immunfluorescence assay. Based on the serologic results, samples were classified as confirmed positive (CP), probable positive (PP), possible other parasitic infection (POPI), and false positive (FP). RESULTS: In 2007 to 2008, a total of 877 of 615,433 donations were discarded due to Chagas assay reactivity. The prevalences (95% confidence intervals [CIs]) among first-time donors for CP, PP, POPI, and FP patterns were 114 (99-129), 26 (19-34), 10 (5-14), and 96 (82-110) per 100,000 donations, respectively. CP and PP had similar patterns of prevalence when analyzed by age, sex, education, and location, suggesting that PP cases represent true T. cruzi infections; in contrast the demographics of donors with POPI were distinct and likely unrelated to Chagas disease. No CP cases were detected among 218,514 repeat donors followed for a total of 718,187 person-years. CONCLUSION: We have proposed a classification algorithm that may have practical importance for donor counseling and epidemiologic analyses of T. cruzi-seroreactive donors. The absence of incident T. cruzi infections is reassuring with respect to risk of window phase infections within Brazil and travel-related infections in nonendemic countries such as the United States.

A inclusão da criança com Síndrome de Down na rede regular de ensino: desafios e possibilidades

O estudo teve como objetivo buscar evidências na literatura acerca da inclusão de crianças com Síndrome de Down na rede regular de ensino. Elaboraram-se revisão da literatura e busca dos artigos nas bases de dados PubMed e PsycINFO, utilizando as palavras-chave Down syndrome, schools, mainstreaming (education), education, infant, newborn, adolescent, child e preschool, no período de 1994 a 2007. Selecionaram-se oito artigos e sua análise permitiu a identificação do tema: experiências e recomendações para a inclusão. Os dados desta revisão, em sua maioria provenientes de relatos de experiências, indicaram que os fatores que colaboraram ou dificultaram o processo de inclusão da criança com síndrome de Down na rede regular de ensino relacionaram-se à escola, aos pais e ao professor. Os resultados deste estudo oferecem possibilidades para melhorar o processo de inclusão, apresentam os desafios e ainda apontam a necessidade do desenvolvimento de novas pesquisas, cujos resultados possam ser aplicados na prática.

Exposições em museus de ciências: reflexões e critérios para análise

Hoje, no que diz respeito à sua interface direta com o público, os museus são considerados, em teoria, instituições com objetivos variados, como educação, lazer, informação e inclusão social. Neste contexto, as exposições aparecem como elemento fundamental da relação entre museus e sociedade, e a expografia torna-se, portanto, ferramenta de comunicação essencial para a efetividade do processo de divulgação. Mas, como se dá a comunicação nestes espaços? Trata-se o presente, de uma reflexão acerca das estratégias de comunicação nas exposições em museus de ciências e sua eficácia. Foram selecionadas três temáticas, a primeira delas é a dos tipos de exposição, e entre as numerosas possibilidades de categorização, optou-se por aquelas descritas por Dean (2003) e Davallon (1986 e 1989). Outra questão discutida é a do uso do objeto, tido, dentre as diferentes mídias a que se pode recorrer em uma exposição, como o diferencial do museu. Por fim, discute-se a questão da interatividade, por vezes reduzida à manipulação, a partir dos níveis propostos por Wagensberg (2000). Apontam-se exemplificações dessas temáticas na exposição de longa duração Pesquisa em Zoologia: a biodiversidade sob o olhar do zoólogo, inaugurada pelo Museu de Zoologia da Universidade de São Paulo em setembro de 2002.

A medição da grandeza practical peak voltage na prática radiológica

OBJETIVO: O objetivo deste trabalho foi estudar a grandeza practical peak voltage (PPV), determinada a partir da forma de onda de tensão aplicada a tubos radiológicos, e compará-la com algumas definições de kVp para diferentes tipos de geradores: monofásico (onda completa, clínico), trifásico (seis pulsos, clínico) e potencial constante (industrial). MATERIAIS E MÉTODOS: O trabalho envolveu a comparação do PPV medido invasivamente (utilizando um divisor de tensão) com a resposta de dois medidores comerciais não invasivos, além dos valores de outras grandezas usadas para medição da tensão de pico aplicada ao tubo de raios X, e a análise da variação do PPV com a ondulação percentual da tensão (ripple). RESULTADOS: Verificou-se que a diferença entre o PPV e as definições mais comuns de tensão de pico aumenta com o ripple. Os valores de PPV variaram em até 3% e 5%, respectivamente, na comparação entre medições invasivas e não invasivas feitas com os equipamentos trifásico e monofásico. CONCLUSÃO: Os resultados demonstraram que a principal grandeza de influência que afeta o PPV é o ripple da tensão. Adicionalmente, valores de PPV obtidos com medidores não invasivos devem ser avaliados considerando que eles dependem da taxa de aquisição e da forma de onda adquirida pelo instrumento.

Declarações de nascidos mortos no município de São Paulo: avaliação descritiva do preenchimento

OBJECTIVES: to study the information contained in Stillbirth Registers (SBRs) in the Municipality of São Paulo. METHODS: the adequacy of the filling out of SBR forms was assessed on the basis of the SBRs (6722) made available by the FSEADE (Foundation for Statistical Data Analysis System), using a Data Completion Index (DCI), making it possible to compare the three years studied (2001-3). Variables relating to the mother and the fetus were included where the DCI was greater than 10%. Education, parity, place of residence, birth type, for the mother and weight, gestational age and underlying cause of death, for the fetus. RESULTS: the absolute stillbirth component changed little in the first two of the three years, falling slightly in the third. The variable most frequently registered was sex (98%), followed by place of residence (82.9%) and parity (70%). The data least often registered were those relating to the mother's age and schooling, 20.0% and 16.7%, respectively. The underlying cause was recorded in 46.7%, fetal weight in 37% and type of birth in 25.3%. CONCLUSIONS: the data demonstrate that the difficulty encountered in incorporating this health indicator into the traditional set of indicators is in part due to the inadequacy of the data provided on the SBR form.

The right to the screen: from media education to educommunication in Brazil

Nowadays in Brazil, some social organizations, governments and mass media are discussing the need to establish an oversight committee to guarantee the quality of television programmes, as well as the need to set a system to determine what kind of pro, gram is appropriate for every television time slot. Across Brazil, a representative body of children and young people have come to the conclusion that the right to receive quality television programmes is not enough. The children of the new generations think they have the right to access new technologies and the production of their own messages, in accordance with their own creativity, interests and lifestyle projects within society.

The right to screens: from media education to educommunication in Brazil

Social organizations, local governments and the media in Brazil are confronting them, selves over the need for an oversight board to guarantee quality television programming, and establishing a system to determine which television programs are appropriate for which television time slots. Across Brazil, a representative body of children and young adults have determined that the right to receive quality programming is not currently being met. Children of the new generation see themselves as having a right to access new technologies which enable them to produce their own messages according to their own creativity, interests, and social participation. This new generation wants to go beyond education in order to watch and conquer their ""right to screens"".

Development, implementation, and assessment of a distance module in endocrine physiology

Rangel EM, Mendes IA, Carnio EC, Marchi Alves LM, Godoy S, Crispim JA. Development, implementation, and assessment of a distance module in endocrine physiology. Adv Physiol Educ 34: 70-74, 2010; doi: 10.1152/advan.00070.2009.-This study aimed to develop, implement, and assess a distance module in endocrine physiology in TelEduc for undergraduate nursing students from a public university in Brazil, with a sample size of 44 students. Stage 1 consisted of the development of the module, through the process of creating a distance course by means of the Web. Stage 2 was the planning of the module's practical functioning, and stage 3 was the planning of student evaluations. In the experts' assessment, the module complied with pedagogical and technical requirements most of the time. In the practical functioning stage, 10 h were dedicated for on-site activities and 10 h for distance activities. Most students (93.2%) were women between 19 and 23 yr of age (75%). The internet was the most used means to remain updated for 23 students (59.0%), and 30 students (68.2%) accessed it from the teaching institution. A personal computer was used by 23 students (56.1%), and most of them (58.1%) learned to use it alone. Access to a forum was more dispersed (variation coefficient: 86.80%) than access to chat (variation coefficient: 65.14%). Average participation was 30 students in forums and 22 students in the chat. Students' final grades in the module averaged 8.5 (SD: 1.2). TelEduc was shown to be efficient in supporting the teaching- learning process of endocrine physiology.

Dendritic Cells Transfected with scFv from Mab 7.B12 Mimicking Original Antigen gp43 Induces Protection against Experimental Paracoccidioidomycosis

Paracoccidioidomycosis (PCM), endemic in Latin America, is a progressive systemic mycosis caused by Paracoccidioides brasiliensis (P. brasiliensis), which primarily attacks lung tissue. Dendritic cells (DCs) are able to initiate a response in naive T cells, and they also participate in Th-cell education. Furthermore, these cells have been used for therapy in several disease models. Here we transfected DCs with a plasmid (pMAC/PS-scFv) encoding a single chain variable fragment (scFv) of an anti-Id antibody that is capable of mimicking gp43, the main antigenic component of P. brasiliensis. First, Balb/c mice were immunized subcutaneously with pMAC/PS-scFv and, after seven days, scFv protein was presented to the regional lymph nodes cells. Moreover, we showed that the DCs transfected with scFv were capable of efficiently activating proliferation of total lymph node cells and inducing a decrease in lung infection. Therefore, our results suggested that the use of scFv-transfected DCs may be a promising therapy in the paracoccidioidomycosis (PCM) model.

The International LAM Registry: A Component of an Innovative Web-Based Clinician, Researcher, and Patient-Driven Rare Disease Research Platform

Background: A relative friability to capture a sufficiently large patient population in any one geographic location has traditionally limited research into rare diseases. Methods and Results: Clinicians interested in the rare disease lymphangioleiomyomatosis (LAM) have worked with the LAM Treatment Alliance, the MIT Media Lab, and Clozure Associates to cooperate in the design of a state-of-the-art data coordination platform that can be used for clinical trials and other research focused on the global LAM patient population. This platform is a component of a set of web-based resources, including a patient self-report data portal, aimed at accelerating research in rare diseases in a rigorous fashion. Conclusions: Collaboration between clinicians, researchers, advocacy groups, and patients can create essential community resource infrastructure to accelerate rare disease research. The International LAM Registry is an example of such an effort.

The characteristics of depressive symptoms in medical students during medical education and training: a cross-sectional study

Background: Medical education and training can contribute to the development of depressive symptoms that might lead to possible academic and professional consequences. We aimed to investigate the characteristics of depressive symptoms among 481 medical students (79.8% of the total who matriculated). Methods: The Beck Depression Inventory (BDI) and cluster analyses were used in order to better describe the characteristics of depressive symptoms. Medical education and training in Brazil is divided into basic (1(st) and 2(nd) years), intermediate (3(rd) and 4(th) years), and internship (5(th) and 6(th) years) periods. The study organized each item from the BDI into the following three clusters: affective, cognitive, and somatic. Statistical analyses were performed using analysis of variance (ANOVA) with post-hoc Tukey corrected for multiple comparisons. Results: There were 184 (38.2%) students with depressive symptoms (BDI > 9). The internship period resulted in the highest BDI scores in comparison to both the basic (p < .001) and intermediate (p < .001) periods. Affective, cognitive, and somatic clusters were significantly higher in the internship period. An exploratory analysis of possible risk factors showed that females (p = .020) not having a parent who practiced medicine (p = .016), and the internship period (p = .001) were factors for the development of depressive symptoms. Conclusion: There is a high prevalence towards depressive symptoms among medical students, particularly females, in the internship level, mainly involving the somatic and affective clusters, and not having a parent who practiced medicine. The active assessment of these students in evaluating their depressive symptoms is important in order to prevent the development of co-morbidities and suicide risk.

Evaluation of gastrointestinal motility in awake rats: a learning exercise for undergraduate biomedical students

Souza MA, Souza MH, Palheta RC Jr, Cruz PR, Medeiros BA, Rola FH, Magalhaes PJ, Troncon LE, Santos AA. Evaluation of gastrointestinal motility in awake rats: a learning exercise for undergraduate biomedical students. Adv Physiol Educ 33: 343-348, 2009; doi: 10.1152/advan.90176.2008.-Current medical curricula devote scarce time for practical activities on digestive physiology, despite frequent misconceptions about dyspepsia and dysmotility phenomena. Thus, we designed a hands-on activity followed by a small-group discussion on gut motility. Male awake rats were randomly submitted to insulin, control, or hypertonic protocols. Insulin and control rats were gavage fed with 5% glucose solution, whereas hypertonic-fed rats were gavage fed with 50% glucose solution. Insulin treatment was performed 30 min before a meal. All meals (1.5 ml) contained an equal mass of phenol red dye. After 10, 15, or 20 min of meal gavage, rats were euthanized. Each subset consisted of six to eight rats. Dye recovery in the stomach and proximal, middle, and distal small intestine was measured by spectrophotometry, a safe and reliable method that can be performed by minimally trained students. In a separate group of rats, we used the same protocols except that the test meal contained (99m)Tc as a marker. Compared with control, the hypertonic meal delayed gastric emptying and gastrointestinal transit, whereas insulinic hypoglycemia accelerated them. The session helped engage our undergraduate students in observing and analyzing gut motor behavior. In conclusion, the fractional dye retention test can be used as a teaching tool to strengthen the understanding of basic physiopathological features of gastrointestinal motility.