Modification of microfiltration membranes by hydrogel impregnation for pDNA purification

The huge efforts for the achievement of highly purified biomolecules are growing every day. A great number of efficient techniques, such as chromatography, are already available in laboratory for separation processes. However, membrane-based technologies are the best match to assure simplicity, efficiency and easy scale-up procedures. Herein we report the modification of a commercial microfiltration membrane for plasmid DNA purification by agarose gel impregnation. The membrane was characterized by SEM, ATR-FTIR, EDS, contact angle, and porosity measurements. Additionally, the membrane pore radius was estimated from observed rejections of different proteins and with that information the rejection of a 6050 bp plasmid DNA (pDNA) molecule was estimated for different values of flux using a theoretical model of large flexible molecules in membranes with parallel cylindrical pores, which is applicable to pDNA ultrafiltration in conventional membranes, as recently shown in the literature. The experimental results show that the modified membrane has higher pDNA rejections than the predicted by the model, suggesting that the different type of porous structure that a hydrogel has, may have a positive effect on pDNA rejections as compared to other biomolecules with more rigid structures, making this type of modified membranes potential better candidates to be used for the selective recovery of pDNA in this type of bioprocesses.

The importance of risk assessment in the context of investment project management: a case study

Risk management is an important component of project management. Nevertheless, such process begins with risk assessment and evaluation. In this research project, a detailed analysis of the methodologies used to treat risks in investment projects adopted by the Banco da Amazonia S.A. was made. Investment projects submitted to the FNO (Constitutional Fund for Financing the North) during 2011 and 2012 were considered for that purpose. It was found that the evaluators of this credit institution use multiple indicators for risk assessment which assume a central role in terms of decision-making and contribute for the approval or the rejection of the submitted projects; namely, the proven ability to pay, the financial records of project promotors, several financial restrictions, level of equity, level of financial indebtedness, evidence of the existence of a consumer market, the proven experience of the partners/owners in the business, environmental aspects, etc. Furthermore, the bank has technological systems to support the risk assessment process, an internal communication system and a unique system for the management of operational risk.

Abordagem histórica e institucional da mudança em contabilidade de gestão: o caso da normalização nos hospitais públicos portugueses

A teoria institucional constituiu o enquadramento no qual foi suportada a pergunta geral desta investigação: como e porquê a Normalização da Contabilidade de Gestão (NCG) nos hospitais públicos portugueses surgiu e evoluiu? O objetivo geral foi compreender de forma profunda o surgimento e a mudança nas regras de NCG dos hospitais públicos portugueses no período histórico 1954-2011. Face ao enquadramento institucional que justificou uma investigação interpretativa, foi usado como método de investigação um estudo de caso explanatório. A evidência sobre o caso da NCG nos hospitais públicos portugueses foi recolhida em documentos e através de 58 entrevistas realizadas em 47 unidades de análise (nos serviços centrais de contabilidade do Ministério da Saúde e em 46 hospitais públicos, num total de 53 existentes). Quanto aos principais resultados obtidos, no período 1954-1974, as regras criadas pelo poder político para controlo dos gastos públicos e a contabilidade orçamental de base de caixa estiveram na génese dos primeiros conceitos de Contabilidade de Gestão (CG) para os serviços públicos de saúde portugueses. A transição de um regime ditatorial para um regime democrático (25 de Abril de 1974), a criação do Plano Oficial de Contabilidade (POC/77) e a implementação de um estado social com Serviço Nacional de Saúde (SNS) criaram a conjuntura crítica necessária para o surgimento de um Plano Oficial de Contabilidade para os Serviços de Saúde (POCSS/80) que incluiu regras de CG. A primeira edição do Plano de Contabilidade Analítica dos Hospitais (PCAH), aprovada em 1996, não foi uma construção de raiz, mas antes uma adaptação para os hospitais das regras de CG incluídas no POCSS/91 que havia revisto o POCSS/80. Após o início da implementação do PCAH, em 1998, ocorreram sequências de autorreforço institucionalizadoras destas normas, no período 1998-2011, por influência de pressões isomórficas coercivas que delinearam um processo de evolução incremental cujo resultado foi uma reprodução por adaptação, num contexto de dependência de recursos. Vários agentes internos e externos pressionaram, no período 2003-2011, através de sequências reativas para a desinstitucionalização do PCAH em resposta ao persistente fenómeno de loose coupling. Mas o PCAH só foi descontinuado nos hospitais com privatização da governação e rejeição dos anteriores sistemas de informação. Ao nível da extensão da teoria, este estudo de caso adotou o institucionalismo histórico na investigação em CG, quanto se sabe pela primeira vez, que se mostra útil na interpretação dos processos e dos resultados da criação e evolução de instituições de CG num determinado contexto histórico. Na condição de dependência de recursos, as sequências de autorreforço, via isomorfismo coercivo, tendem para uma institucionalização com fenómeno de loose coupling. Como resposta a este fenómeno, ocorrem sequências reativas no sentido da desinstitucionalização. Perante as pressões (políticas, funcionais, sociais e tecnológicas) desinstitucionalizadoras, o fator governação privada acelera o processo de desinstitucionalização, enquanto o fator governação pública impede ou abranda esse processo.

Pesquisa participativa: novas vias de estudo da relação entre jornalismo e democracia

É difícil conseguir captar a essência das dietas mediáticas e do pensamento dos cidadãos sobre a democracia, ido além da visão mais tradicional, associada a um pensamento de sentido negativo e de afastamento. Neste artigo indicamos as vantagens de optar por metodologias qualitativas participativas e longitudinais, que permitem melhor apreender atitudes e hábitos e os contextos do quotidiano em que se desenvolvem. Estas opções possibilitam melhor conhecer espaços de micro análise e ainda favorecer ambientes em que os participantes na pesquisa sentem que dão um contributo ativo para o desenrolar da investigação. Por esta via, consegue-se apreender melhor processos de ligação entre jornalismo e democracia, tendo uma das conclusões deste estudo apontado para processos de ligação próxima entre opiniões sobre jornalismo, democracia e contextos quotidianos. Os jovens que indicaram ter uma visão mais alargada sobre o jornalismo, designadamente percepcionando as suas ambivalências e considerando-as inerentes à profissão, foram também os que revelaram maior capacidade para entender a política numa visão alargada do seu significado.

Permanent and temporary pacemaker implantation after orthotopic heart transplantation

PURPOSE:To determine the indication for and incidence and evolution of temporary and permanent pacemaker implantation in cardiac transplant recipients. METHODS: A retrospective review of 114 patients who underwent orthotopic heart transplantation InCor (Heart Institute USP BR) between March 1985 and May 1993. We studied the incidence of and indication for temporary pacing, the relationship between pacing and rejection, the need for pemanent pacing and the clinical follow-up. RESULTS: Fourteen of 114 (12%)heart transplant recipients required temporary pacing and 4 of 114 (3.5%) patients required permanent pacing. The indication for temporary pacing was sinus node dysfunction in 11 patients (78.5%) and atrioventricular (AV) block in 3 patients (21.4%). The indication for permanent pacemaker implantation was sinus node dysfunction in 3 patients (75%) and atrioventricular (AV) block in 1 patient (25%). We observed rejection in 3 patients (21.4%) who required temporary pacing and in 2 patients (50%) who required permanent pacing. The previous use of amiodarone was observed in 10 patients (71.4%) with temporary pacing. Seven of the 14 patients (50%) died during follow-up. CONCLUSION: Sinus node dysfunction was the principal indication for temporary and permanent pacemaker implantation in cardiac transplant recipients. The need for pacing was related to worse prognosis after cardiac transplantation.

Heart transplantation in neonates and children. Intermediate-term results

OBJECTIVE: To assess intermediate-term outcome in children who have undergone orthotopic heart transplantation. METHODS: We carried out a longitudinal and prospective study between October '92 and June '99 comprising 20 patients with ages ranging from 12 days to 7 years (mean of 2.8 years). We employed a double immunosuppression protocol with cyclosporine and azathioprine and induction therapy with polyclonal antithymocyte serum. Survival and complications resulting from the immunosuppression protocol were analyzed. RESULTS:The double immunosuppression protocol and the induction therapy with polyclonal antithymocyte serum resulted in an actuarial survival curve of 90% and 78.2% at 1 and 6 years, respectively, with a mean follow-up period of 3.6 years. One patient died due to acute rejection 40 days after transplantation; another patient died 2 years after transplantation due to lymphoproliferative disorder; a third patient died because of primary failure of the graft; and a fourth patient died due to bronchopneumonia. The major complications were as follows: acute rejection, infection, nephrotoxicity, and systemic hypertension. The means of rejection and infection episodes per patient were 2.9 and 3.4, respectively. After one year of transplantation, a slight reduction in the creatinine clearance and systemic hypertension were observed in 7 (38.9%) patients. CONCLUSION: Heart transplantation made life possible for those patients with complex congenital heart diseases and cardiomyopathies in refractory congestive heart failure constituting a therapeutical option for this group of patients in the terminal phase.

Modelos de gestão pública e as opções de reforma do XIX governo constitucional: análise do discurso

Dissertação de Mestrado em Gestão e Políticas Públicas

Fair procedures. Evidence from games involving lotteries

Procedural fairness plays a prominent role in the social discourse concerning the marketplace in particular, and social institutions in general. Random procedures are a simple case, and they have found application in several important social allocation decisions. We investigate random procedures in the laboratory. We find that an unbiased random procedure is an acceptable substitute for an unbiased allocation: similar patterns of acceptance and rejection result when either is inserted as a feasible proposal in a sequential battle-of-the-sexes. We also find that unbiasedness, known to be a crucial characteristic of allocation fairness, is important to procedural fairness: in the context of a random offer game, a biased outcome is more readily accepted when chosen by an unbiased random draw than by one that is biased. Procedural fairness is conceptually different than allocation fairness or attribution-based behavior, and none of the current models of fairness and reciprocity captures our results. Post hoc extension of one of these models (ERC) suggests that a deeper understanding of procedural fairness requires further investigation of competing fairness norms.

The Rise of democracy in Europe and the fight against mass poverty in Latin America: the implications for marxist thought of some recent mainstream papers

Recently a number of mainstream papers have treated the rise of democracy in 19th century Europe and its instability in Latin America in an eminently Marxist fashion. This paper sets out their implications for Marxist thought. With respect to Europe, Marx's emphasis on political action backed by the threat of violence is vindicated but his justification for socialism is not. With respect to Latin America, the unequal distribution of wealth is the cause of political instability that is, in turn, the root cause of mass poverty. In addition it is possible to explain some of the paradoxical characteristics of neo-liberalism and to make a weak argument for socialism in spite of its rejection in Europe.

Chagas' disease: selective affinity and cytotoxicity of Trypanosoma cruzi-immune lymphocytes to parasympathetic ganglion cells

The megaesophagus and megacolon endemic in South America are related , to Chagas' disease. These mega conditions are found in patients with chronic Chagas's infection, when the parasite is not demonstrable in the lesions. These are characterized by depopulation of parasympathetic ganglion cells, dilation and hypertrophy of the viscera. In the experiments described here we deminstrate a selective affinity and adherence of Trypanosoma cruzi-immune lymphocytes to myenteric, parasympathetic ganglion cells, leading to neuronolysis. None of these features are observed when non-immune lymphocytes from control rabbits are used, or when the immune lymphocytes are allowed to react with CNS neurons. This demonstration is an indication of the high degree of specificity of the destruction of parasympathetic neurons in Chagas' disease. We postulate that the T. cruzi-immune lymphocyte rejection of parasympathetic neurons, but not of CNS neurons, might be related to recognition of a cross-reacting antigenic determinant secreted only by the target neurons. In favor of this interpretation is the observation of lymphocytic infiltrates and parasympathetic ganglion cell destruction in chronic Chagas' infection in the absence of encephalitis.

Diseño de un diplexor basado en resonadores BAW

Este proyecto plantea el diseño de un diplexor basado en resonadores BAW, pensado para aplicaciones de comunicaciones móviles. Éste permite a los terminales seleccionar los canales de uplink y downlink en sistemas que utilicen duplexado por división en frecuencia. Se plantean tres alternativas de diseño a partir de filtros implementados con resonadores BAW en una topología en escalera. Éstas serán evaluadas y comparadas considerando las pérdidas de inserción, el rechazo fuera de banda y las dimensiones de la estructura obtenida.

Prevention of bone marrow graft failure by an anti LFA-1 monoclonal antibody

Graft rejection is the major cause of failure of HLA mismatched bone marrow transplantation because of residual host immunity. we have proposed to use a monoclonal murine antibody specific for the LFA-1 molecule (25-3) to prevent graft failure in HLA mismatched bone marrow transplantation (BMT). The rationale for this approach is three fold: LFA-1 deficient patients (3/3) do not reject HLA mismatched BMT; anti LFA-1 blocka in vitro the induction of T cell responses and T/ non T cytotoxic functions; LFA-1 is not expressed by other cells than leucocytes. We have accordingly treated twenty two patients with inherited diseases and 8 with leikemia. The bone marrow was T cells depled by E rosetting of Campath antibody. The antibody was given at days -3, -1, +1, +3, +5 at dose of .1 mg/kg/d for the first 9 and then .2mg/kg/d from day -3 to +6. Engraftment occured in 23/30 patients as shown by at least HLA typing. Hematological recovery was rapid, GVH was limited. Side effects of antibody infusion included fever and possibly an increased incidence of early bacteral infection (sepsis, 1 death). Immunological reconstitution occured slowly leading in six cases to EBV-induced B cell poliferation (1 death and in two others to transient auto immune hemolytic anemia. There has been only one secondary graft rejection. Sisteen patients are alive 3 to 26 months post transplant with functional grafts. Although the number of patients treated is still low the absence of late rejection so far, gives hope for long term maintenance of the graft using anti LFA-1. Since the antibody is an IgG 1 unable to bind human complement, and since it is known to inhibit phagocytosis, there is a good suggestion that 25-3 act through functional blocking of host T and non T luymphocytes at both induction and effector levels.

Price level convergence, purchasing power parity and multiple structural breaks: an application to US cities

This article provides a fresh methodological and empirical approach for assessing price level convergence and its relation to purchasing power parity (PPP) using annual price data for seventeen US cities. We suggest a new procedure that can handle a wide range of PPP concepts in the presence of multiple structural breaks using all possible pairs of real exchange rates. To deal with cross-sectional dependence, we use both cross-sectional demeaned data and a parametric bootstrap approach. In general, we find more evidence for stationarity when the parity restriction is not imposed, while imposing parity restriction provides leads toward the rejection of the panel stationar- ity. Our results can be embedded on the view of the Balassa-Samuelson approach, but where the slope of the time trend is allowed to change in the long-run. The median half-life point estimate are found to be lower than the consensus view regardless of the parity restriction.

Switch to Sirolimus-based immunosuppresion in stable renal transplant recipients

Purpose: Sirolimus (SRL) has been used to replace calcineurin inhibitors (CNI) for various indications including CNI-induced toxicity. The aim of this study was to evaluate the efficacy and safety of switching from CNI to SRL in stable renal transplant recipients (RTR) with low grade proteinuria (<1 g/24 h). Methods and materials: Between 2001 and 2007, 41 patients (20 females, 21 males; mean age 47 ± 13) were switched after a median time post-transplantation of 73.5 months (range 0.2-273.2 months). Indications for switch were CNI nephrotoxicity (39%), thrombotic micro-angiopathy (14.6%), post-transplantation cancer (24.4%), CNI neurotoxicity (7.4%), or others (14.6%). Mean follow-up after SRL switch was 23.8±16.3 months. Mean SRL dosage and through levels were 2.4 ± 1.1 mg/day and 8 ± 2.2 ug/l respectively. Immunosuppressive regiments were SRL + mycophenolate mofetil (MMF) (31.7%), SRL + MMF + prednisone (36.58%), SRL + prednisone (19.51%), SRL + Azathioprine (9.75%), or SRL alone (2.43%). Results: Mean creatinine decreased from 164 to 143 μmol/l (p <0.03), mean estimated glomerular filtration rate (eGFR) increased significantly from 50.13 to 55.01 ml/minute (p <0.00001), mean systolic and diastolic blood pressure decreased from 138 to 132 mm Hg (p <0.03) and from 83 to78 mm Hg (p <0.01), but mean proteinuria increased from 0.21 to 0.63 g/24 h (p <0.001). While mean total cholesterolemia didn't increased significantly from 5.09 to 5.56 mmol/l (p = 0.06). The main complications after SRL switch were dermatitis (19.5%), urinary tract infections (24.4%), ankle edema (13.3%), and transient oral ulcers (20%). Acute rejection after the switch occurred in 7.3% of patients (n = 3), and 2 acute rejections were successfully treated with corticosteroids and 1 did not respond to treatment (not related to switch). SRL had to be discontinued in 17% of patients (2 nephrotic syndromes, 2 severe edema, 1 acute rejection, 1 thrombotic micro-angiopathy, and 1 fever). Conclusion: In conclusion, we found that switching from CNI to SRL in stable RTR was safe and associated with a significant improvement of renal function and blood pressure. Known side-effects of SRL led to drug discontinuation in less than 20% of patients and the acute rejection rate was 7.3%. This experience underlines the importance of patient selection before switching to SRL, in particular regarding preswitch proteinuria.

Oral valganciclovir prophylaxis in kidney transplant recipients

Background: Immunosuppressive and antivira[ prophy[ actic drugs are needed to prevent acute rejection and infection after organ transplantation. We assessed the effectiveness of a new combined regimen introduced at our transplantation center. Methods: We reviewed at[ consecutive patients who underwent kidney transplantation at our institution over a 5.5-year period, with a follow-up of at [east 6 months. Patients transplanted from 1/2000 to 3/2003 (Period 1) were compared to patients transplanted from 4/2003 to 7/2005 (Period 2). In period 1, patients were treated with Basi[iximab, Cic[osporin, steroids and Mycophenotate or Azathioprine. Prophylaxis with Va[acic[ ovir was prescribed in CMV D+/R- patients; otherwise, a preemptive antivira[ approach was used. In period 2, immunosuppressive drugs were Basi[- iximab, Tacro[imus, steroids and Mycopheno[ate. A 3-month CMV prophylaxis with Va[gancic[ovir was used, except in D-/R- patients. Results: Sixty-three patients were transplanted in period 1 and 70 patients in period 2. Baseline characteristics of both groups were comparable; in particular 17% of patients were CMV D+/R- in period 1 compared to 23% in period 2 (p=0.67). Acute rejection was more frequent in period 1 than in period 2 (40% of patients vs 7%, respectively p<0.001). Nineteen patients (30%) in period 1 were diagnosed with CMV infection/disease that required treatment, compared with 8 patients (11.4%) in period 2 (p = 0.003). Of these 8 patients, at[ had CMV infection/disease after discontinuation of Va[gancic[ovir prophylaxis, 6 were D+/R- (75%), and at[ were treated with oral Va[gancic[ovir. There was no difference between periods in terms of incidence of BK nephropathy, post-transplant [ymphopro[ iferative disease, graft toss, and mortality. Conclusions: These results indicate that a 3-month course of oral Va[gancic[ovir is very effective to prevent CMV infection/disease in kidney transplantation. Late-onset CMV disease is a residual problem in D+/R- patients receiving VGC prophylaxis.