Selective Permeation and Organic Extraction of Recombinant Green Fluorescent Protein (gfpuv) from Escherichia coli

Background Transformed cells of Escherichia coli DH5-α with pGFPuv, induced by IPTG (isopropyl-β-d-thiogalactopyranoside), express the green fluorescent protein (gfpuv) during growth phases. E. coli subjected to the combination of selective permeation by freezing/thawing/sonication cycles followed by the three-phase partitioning extraction (TPP) method were compared to the direct application of TPP to the same culture of E. coli on releasing gfpuv from the over-expressing cells. Material and Methods Cultures (37°C/100 rpm/ 24 h; μ = 0.99 h-1 - 1.10 h-1) of transformed (pGFP) Escherichia coli DH5-α, expressing the green fluorescent protein (gfpuv, absorbance at 394 nm and emission at 509 nm) were sonicated in successive intervals of sonication (25 vibrations/pulse) to determine the maximum amount of gfpuv released from the cells. For selective permeation, the transformed previously frozen (-75°C) cells were subjected to three freeze/thaw (-20°C/ 0.83°C/min) cycles interlaid by sonication (3 pulses/ 6 seconds/ 25 vibrations). The intracellular permeate with gfpuv in extraction buffer (TE) solution (25 mM Tris-HCl, pH 8.0, 1 mM β-mercaptoethanol β-ME, 0.1 mM PMSF) was subjected to the three-phase partitioning (TPP) method with t-butanol and 1.6 M ammonium sulfate. Sonication efficiency was verified on the application to the cells previously treated by the TPP method. The intra-cell releases were mixed and eluted through methyl HIC column with a buffer solution (10 mM Tris-HCl, 10 mM EDTA, pH 8.0). Results The sonication maximum released amount obtained from the cells was 327.67 μg gfpuv/mL (20.73 μg gfpuv/mg total proteins – BSA), after 9 min of treatment. Through the selective permeation by three repeated freezing/thawing/sonication cycles applied to the cells, a close content of 241.19 μg gfpuv/mL (29.74 μg gfpuv/mg BSA) was obtained. The specific mass range of gfpuv released from the same cultures, by the three-phase partitioning (TPP) method, in relation to total proteins, was higher, between 107.28 μg/mg and 135.10 μg/mg. Conclusions The selective permeation of gfpuv by freezing/thawing/sonication followed by TPP separation method was equivalent to the amount of gfpuv extracted from the cells directly by TPP; although selective permeation extracts showed better elution through the HIC column.

Management of diabetes mellitus and associated cardiovascular risk factors in Brazil – the Brazilian study on the practice of diabetes care

Abstract Background The Brazilian Study on the Practice of Diabetes Care main objective was to provide an epidemiological profile of individuals with type 1 and 2 diabetes mellitus (DM) in Brazil, concerning therapy and adherence to international guidelines in the medical practice. Methods This observational, cross-sectional, multicenter study collected and analyzed data from individuals with type 1 and 2 DM attending public or private clinics in Brazil. Each investigator included the first 10 patients with type 2 DM who visited his/her office, and the first 5 patients with type 1 DM. Results A total of 1,358 patients were analyzed; 375 (27.6%) had type 1 and 983 (72.4%) had type 2 DM. Most individuals were women, Caucasian, and private health care users. High prevalence rates of hypertension, dyslipidemia and central obesity were observed, particularly in type 2 DM. Only 7.3% and 5.1% of the individuals with types 1 and 2 DM, respectively, had optimal control of blood pressure, plasma glucose and lipids. The absence of hypertension and female sex were associated with better control of type 1 DM and other cardiovascular risk factors. In type 2 DM, older age was also associated with better control. Conclusions Female sex, older age, and absence of hypertension were associated with better metabolic control. An optimal control of plasma glucose and other cardiovascular risk factors are obtained only in a minority of individuals with diabetes. Local numbers, compared to those from other countries are worse.

Alterações de parâmetros relacionados ao metabolismo ósseo em mulheres submetidas à derivação gástrica em Y de Roux

OBJETIVO: Avaliar o metabolismo ósseo e a densidade mineral óssea (DMO) em mulheres adultas pós-derivação gástrica em Y de Roux (DGYR). SUJEITOS E MÉTODOS: Estudo transversal com 48 mulheres submetidas a DGYR há três anos e 41 saudáveis. Dados obtidos: índice de massa corporal (IMC), atividade física, consumo alimentar e DMO da coluna lombar, colo e fêmur total. Dosagem de cálcio, fósforo, magnésio, albumina, fosfatase alcalina, telopeptídeo-C (CTX), paratormônio (PTH), 25-hidroxivitamina D (25OHD), osteocalcina e cálcio urinário. RESULTADOS: Maiores alterações no grupo DGYR observadas nos níveis de osteocalcina (p < 0,001), CTX (p < 0,001) e PTH (p < 0,001). Deficiência de 25OHD foi a mais frequente no grupo DGYR (p = 0,010). Deficiência/insuficiência de 25OHD associou-se com hiperparatiroidismo secundário (p = 0,025). Não houve diferença entre os grupos em relação à DMO. A ingestão de energia (p = 0,036) e proteína (p = 0,004) foi maior no grupo controle. CONCLUSÃO: Em mulheres pós-DGYR, encontraram-se alta frequência de deficiência de vitamina D, hiperparatireoidismo secundário e elevação nos marcadores de remodelação óssea, sem alteração na DMO quando comparado com o grupo controle não obeso.

Revista do Museu Paulista e(m) capas: identidade e representação institucional em texto e imagem

Este artigo propõe reflexão sobre a escrita da história, a partir de estudo das capas da Revista do Museu Paulista, especialmente durante as gestões dos dois primeiros diretores da instituição, quando certa continuidade editorial foi preservada. Tomadas como "fontes", ou seja, objeto de trabalho do historiador, as capas revelam vestígios de processos que permitem reconstruir, em suas dimensões históricas, as complexas imbricações entre a produção da Revista, sua materialidade, e os atos de leitura e interpretação que condicionam e refletem relações entre produtores de textos e seus leitores nas primeiras décadas do século XX, a partir de São Paulo, onde era editada.

Biological and physicochemical stability of ceftazidime and aminophylline on glucose parenteral solution

Ceftazidime is a broad spectrum antibiotic administered mainly by the parenteral route, and it is especially effective against Pseudomonas aeruginosa. The period of time in which serum levels exceed the Minimum Inhibitory Concentration (MIC) is an important pharmacodynamic parameter for its efficacy. One of the forms to extend this period is to administer the antibiotic by continuous infusion, after prior dilution in a Parenteral Solution (PS). The present work assessed the stability of ceftazidime in 5% glucose PS for 24 hours, combined or not with aminophylline, through High Performance Liquid Chromatography (HPLC). The physicochemical evaluation was accompanied by in vitro antimicrobial activity compared MIC test in the 24-hour period. Escherichia coli and Pseudomonas aeruginosa were the microorganisms chosen for the MIC comparison. The HPLC analysis confirmed ceftazidime and aminophylline individual stability on PS, while the MIC values were slightly higher than the mean described in the literature. When both drugs were associated in the same PS, the ceftazidime concentration by HPLC decreased 25% after 24 hours. Not only did the MIC values show high loss of antibiotic activity within the same period, but also altered MIC values immediately after the preparation, which was not detected by HPLC. Our results indicate that this drug combination is not compatible, even if used right away, and that PS might not be the best vehicle for ceftazidime, emphasizing the importance of the MIC evaluation for drug interactions.

Microbial Translocation Is Associated with Extensive Immune Activation in Dengue Virus Infected Patients with Severe Disease

Background: Severe dengue virus (DENV) disease is associated with extensive immune activation, characterized by a cytokine storm. Previously, elevated lipopolysaccharide (LPS) levels in dengue were found to correlate with clinical disease severity. In the present cross-sectional study we identified markers of microbial translocation and immune activation, which are associated with severe manifestations of DENV infection. Methods: Serum samples from DENV-infected patients were collected during the outbreak in 2010 in the State of Sa˜o Paulo, Brazil. Levels of LPS, lipopolysaccharide binding protein (LBP), soluble CD14 (sCD14) and IgM and IgG endotoxin core antibodies were determined by ELISA. Thirty cytokines were quantified using a multiplex luminex system. Patients were classified according to the 2009 WHO classification and the occurrence of plasma leakage/shock and hemorrhage. Moreover, a (non-supervised) cluster analysis based on the expression of the quantified cytokines was applied to identify groups of patients with similar cytokine profiles. Markers of microbial translocation were linked to groups with similar clinical disease severity and clusters with similar cytokine profiles. Results: Cluster analysis indicated that LPS levels were significantly increased in patients with a profound pro-inflammatory cytokine profile. LBP and sCD14 showed significantly increased levels in patients with severe disease in the clinical classification and in patients with severe inflammation in the cluster analysis. With both the clinical classification and the cluster analysis, levels of IL-6, IL-8, sIL-2R, MCP-1, RANTES, HGF, G-CSF and EGF were associated with severe disease. Conclusions: The present study provides evidence that both microbial translocation and extensive immune activation occur during severe DENV infection and may play an important role in the pathogenesis.

Biological and physicochemical stability of ceftazidime and aminophylline on glucose parenteral solution

Ceftazidime is a broad spectrum antibiotic administered mainly by the parenteral route, and it is especially effective against Pseudomonas aeruginosa. The period of time in which serum levels exceed the Minimum Inhibitory Concentration (MIC) is an important pharmacodynamic parameter for its efficacy. One of the forms to extend this period is to administer the antibiotic by continuous infusion, after prior dilution in a Parenteral Solution (PS). The present work assessed the stability of ceftazidime in 5% glucose PS for 24 hours, combined or not with aminophylline, through High Performance Liquid Chromatography (HPLC). The physicochemical evaluation was accompanied by in vitro antimicrobial activity compared MIC test in the 24-hour period. Escherichia coli and Pseudomonas aeruginosa were the microorganisms chosen for the MIC comparison. The HPLC analysis confirmed ceftazidime and aminophylline individual stability on PS, while the MIC values were slightly higher than the mean described in the literature. When both drugs were associated in the same PS, the ceftazidime concentration by HPLC decreased 25% after 24 hours. Not only did the MIC values show high loss of antibiotic activity within the same period, but also altered MIC values immediately after the preparation, which was not detected by HPLC. Our results indicate that this drug combination is not compatible, even if used right away, and that PS might not be the best vehicle for ceftazidime, emphasizing the importance of the MIC evaluation for drug interactions.

Una realizzazione distribuita di test neuropsicologici

Questa tesi fornisce una realizzazione di alcuni test neuropsicologici, scelti in collaborazione di un neuropsicologo, che cercano di sostituire foglio e penna con touch screen e dita. L'obiettivo è quello di ricreare il test senza aggiungere agenti esterni derivanti dalla nuova interfaccia, per far sì che la fruizione del test sia buona almeno quanto quella del corrispettivo non digitale. Il risultato finale dovrebbe apportare dei vantaggi sostanziali al processo di somministrazione di questi test.

Come tradurre il metadiscorso letterario. Esempi di scrittura femminile nell'Ottocento austriaco

L’argomento affrontato nel presente lavoro di tesi dal titolo “Come tradurre il metadiscorso letterario. Esempi di scrittura femminile nell’Ottocento austriaco” è la versione interlinguistica di testi saggistici afferenti all’ambito del metadiscorso letterario. Nello specifico, non vengono analizzati testi di critica e/o metodologia ma scritti funzionali, di forte carattere pragmatico, che pur tuttavia rientrano tra le testimonianze di alta caratura letteraria, perché dovuti ad autrici che hanno fatto dell’espressione estetica la propria finalità primaria. I materiali scelti per l’analisi linguistico-testuale, compresi in un arco temporale tra la fine del Settecento e la metà dell’Ottocento, sono realizzati da donne che hanno operato in ambito teatrale facendo dell’attività di scrittura lo strumento della propria emancipazione intellettuale ed economica. La necessità di trovare una via alla pubblicazione le ha indotte a strategie di scrittura connotate da particolari stilemi e artifici retorici atti a favorire l’accettazione e la diffusione delle proposte editoriali di cui questi “paratesti” costituivano il momento giustificante. Il “lavoro di penna” è un’esperienza che viene ad assumere molteplici contorni, non privi di ricadute al momento della scelta delle strategie traduttive. Dal punto di vista formale, le testimonianze si collocano in una zona di modalità espressiva contigua alla testimonianza autobiografica. Il periodo storico e l’area di provenienza delle autrici hanno reso necessario un approccio capace di incrociare il piano diacronico con la dimensione diatopica, rendendo conto delle componenti diamesiche di una scrittura che nasce dal teatro per il teatro e ad esso e ai suoi frequentatori deve rapportarsi. Il modello traduttologico applicato ricava le sue linee fondamentali dalle riflessioni della linguistica testuale e dall’approccio integrato/multidisciplinare della “prototipologia dinamica”.

Un'applicazione dei web playground all'insegnamento dell'informatica

Lo scopo di questa tesi è dimostrare che è possibile effettuare in maniera proficua esami di tecnologie web all'università senza utilizzare carta e penna. Nello specifico si parla di ExaM Bin, piattaforma open-source creata come adattamento del tool di web playground JSBIN per fare esami di tecnologie web.

Policy Contexts of Social Work in Britain: the wider implications of 'New' Labour and the 'New Legal Regime'

Several commentators have expressed disappointment with New Labour's apparent adherence to the policy frameworks of the previous Conservative administrations. The employment orientation of its welfare programmes, the contradictory nature of the social exclusion initiatives, and the continuing obsession with public sector marketisation, inspections, audits, standards and so on, have all come under critical scrutiny (c.f., Blyth 2001; Jordan 2001; Orme 2001). This paper suggests that in order to understand the socio-economic and political contexts affecting social work we need to examine the relationship between New Labour's modernisation project and its insertion within an architecture of global governance. In particular, membership of the European Union (EU), International Monetary Fund (IMF) and World Trade Organisation (WTO) set the parameters for domestic policy in important ways. Whilst much has been written about the economic dimensions of 'globalisation' in relation to social work rather less has been noted about the ways in which domestic policy agenda are driven by multilateral governance objectives. This policy dimension is important in trying to respond to various changes affecting social work as a professional activity. What is possible, what is encouraged, how things might be done, is tightly bounded by the policy frameworks governing practice and affected by those governing the lives of service users. It is unhelpful to see policy formulation in purely national terms as the UK is inserted into a network governance structure, a regulatory framework where decisions are made by many countries and organisations and agencies. Together, they are producing a 'new legal regime', characterised by a marked neo-liberal policy agenda. This paper aims to demonstrate the relationship of New Labour's modernisation programme to these new forms of legality by examining two main policy areas and the welfare implications they are enmeshed in. The first is privatisation, and the second is social policy in the European Union. Examining these areas allows a demonstration of how much of the New Labour programme can be understood as a local implementation of a transnational strategy, how parts of that strategy produce much of the social exclusion it purports to address, and how social welfare, and particularly social work, are noticeable by their absence within policy discourses of the strategy. The paper details how the privatisation programme is considered to be a crucial vehicle for the further development of a transnational political-economy, where capital accumulation has been redefined as 'welfare'. In this development, frameworks, codes and standards are central, and the final section of the paper examines how the modernisation strategy of the European Union depends upon social policy marked by an employment orientation and risk rationality, aimed at reconfiguring citizen identities.The strategy is governed through an 'open mode of coordination', in which codes, standards, benchmarks and so on play an important role. The paper considers the modernisation strategy and new legality within which it is embedded as dependent upon social policy as a technology of liberal governance, one demonstrating a new rationality in comparison to that governing post-Second World War welfare, and which aims to reconfigure institutional infrastructure and citizen identity.

What Price Social and Health Care? Commodities, Competition and Consumers

In this paper we explore some important disputes and problems surrounding the legal status and social purpose of Health