In vitro susceptibility testing of Fonsecaea pedrosoi to antifungals

Based on the difficulties experienced in the treatment of chromoblastomycosis, 12 primary human isolates of F. pedrosoi, were tested for their in vitro susceptibility to various antimycotics. We adapted the recommendations of the NCCLS for yeasts and followed the indications for mold testing from other authors in order to determine their MIC’s and the MLC’s. It was found that a significant proportion of the isolates were resistant to 3 of the 4 antimycotics tested, as revealed by high MIC values, as follows: 33% were resistant to amphotericin B (AMB), 58.3% to 5 fluocytosine (5 FC) and 66.7% to fluconazole (FLU). Contrarywise, none of the isolates proved resistant to itraconazole (ITZ). Determination of the MLC’s revealed that a larger proportion of the isolates were not killed by AMB, 5 FC (91.7%), FLU (100%) or even, ITZ (41.7%). These data indicate that it would be desirable to determine the susceptibility of F. pedrosoi before initiating therapy, in order to choose the more effective antifungal and avoid clinical failure

Massive shift in the pneumococcal nasopharyngeal flora after the 7-valent conjugate vaccine: epidemiological studies and testing pathogenic potential in animal models

Dissertation presented to obtain the PhD degree in Biology/Molecular Biology by Universidade Nova de Lisboa, Instituto de Tecnologia Química e Biológica

Congenital cytomegalovirus infection in a neonatal intensive care unit in Brazil evaluated by PCR and association with perinatal aspects

Cytomegalovirus (CMV) infection is the most common congenital infection, affecting 0.4% to 2.3% newborns. Most of them are asymptomatic at birth, but later 10% develop handicaps, mainly neurological disturbances. Our aim was to determine the prevalence of CMV shed in urine of newborns from a neonatal intensive care unit using the polymerase chain reaction (PCR) and correlate positive cases to some perinatal aspects. Urine samples obtained at first week of life were processed according to a PCR protocol. Perinatal data were collected retrospectively from medical records. Twenty of the 292 cases (6.8%) were CMV-DNA positive. There was no statistical difference between newborns with and without CMV congenital infection concerning birth weight (p=0.11), gestational age (p=0.11), Apgar scores in the first and fifth minutes of life (p=0.99 and 0.16), mother's age (p=0.67) and gestational history. Moreover, CMV congenital infection was neither related to gender (p=0.55) nor to low weight (<2,500g) at birth (p=0.13). This high prevalence of CMV congenital infection (6.8%) could be due to the high sensitivity of PCR technique, the low socioeconomic level of studied population or the severe clinical status of these newborns.

Dor crónica lombar : incapacidada auto-reportada em trabalhadores de saúde e sua relação com variáveis psicossociais e intensidade da dor

RESUMO: A dor crónica lombar, é uma condição de saúde cuja prevalência tem aumentado nas últimas décadas. É uma condição que pode ser bastante incapacitante para o indivíduo e por consequência, ter importante impacto social e económico na sociedade. É um fenómeno complexo, multifactorial e pouco estudado na população portuguesa. Objectivo: Estudar a associação entre a catastrofização da dor, crenças de medo evitamento da dor, intensidade da dor e a incapacidade funcional auto reportada em indivíduos com dor crónica lombar. Metodologia: Estudo observacional analítico de corte transversal, com uma amostra de 38 indivíduos com dor crónica lombar, seleccionados a partir de uma população de 186 trabalhadores de uma unidade local de saúde. A recolha de dados foi realizada através de 4 instrumentos de avaliação: Questionário de caracterização e levantamento de factores de risco e impacto associados à dor crónica lombar; Questionário de incapacidade de Roland e Morris; Escala de catastrofização da dor; e Questionário de crenças de medo evitamento da dor. A análise dos dados foi feita através de estatística descritiva pela distribuição de frequências e medidas de tendência central para análise da prevalência e caracterização da amostra e por estatística inferencial para estudar as relações entre variáveis através do teste de correlação não paramétrico de Spearman. Resultados: A variável catastrofização da dor obteve um valor de correlação com a incapacidade auto-reportada de rs=0,473, para p<0,01; a variável crença de medo evitamento da dor relacionada com o trabalho obteve um valor de correlação com a incapacidade auto-reportada de rs=0,462 para p<0,01, a percepção da intensidade actual de dor e a intensidade percepcionada no ano anterior, obtiveram valores de correlação com a incapacidade auto-reportada de rs=0,327 e rs= 0,359 respectivamente para valor de p<0,05. Conclusão: As variáveis psicossociais catastrofização da dor e crença de medo evitamento da dor relacionada com o trabalho, influenciam de forma moderada a incapacidade em indivíduos com dor crónica lombar. A associação entre a intensidade da dor e a incapacidade parece ter um papel menos importante demonstrando associações baixas.--------------------------ABSTRACT: Chronic low back pain is a health condition whose prevalence has increased in recent decades. It is a condition that can be quite disabling for the individual and therefore have important social and economic impact on society. It is a complex phenomenon, multifactorial and poorly studied in the Portuguese population. Objective: To study the association between pain catastrophizing, fear avoidance beliefs, pain, pain intensity and self-reported functional disability in individuals with chronic low back pain. Methods: Observational analytical cross sectional study of a sample of 38 individuals with chronic low back pain, selected from a population of 186 workers at a local health unit. Data collection was performed through four assessment instruments: questionnaire characterization, evaluation of risk factors and impact associated to chronic low back pain, questionnaire Roland and Morris disability, pain catastrophizing scale and fear avoidance beliefs questionnaire. Data analysis was performed using descriptive statistics for the distribution of frequencies and measures of central tendency to analyze the prevalence and characteristics of the sample and inferential statistics to study the relationships between variables by testing for Spearman nonparametric correlation. Results: The pain catastrophizing variable had a correlation value rs= 0,473, p<0,01 with the self-reported disability, the variable of fear avoidance belief of pain related to the work achived a correlation value with the self-reported disability, rs = 0.462 p <0.01, current pain intensity and in the previous year obtained values of correlation with self-reported disability rs = 0.327 and rs = 0.359 respectively for values of p <0.05 .Conclusion: The psychosocial variables of pain catastrophizing and fear avoidance belief of pain related to the work had a moderate association with disability in individuals with chronic low back pain. The association between pain intensity and disability seems to have a less important role demonstrating low associations.

Evaluation of Etest and macrodilution broth method for antifungal susceptibility testing of Candida sp strains isolated from oral cavities of AIDS patients

A comparison of the Etest and the reference broth macrodilution susceptibility test for fluconazole, ketoconazole, itraconazole and amphotericin B was performed with 59 of Candida species isolated from the oral cavities of AIDS patients. The Etest method was performed according to the manufacturer's instructions, and the reference method was performed according to National Committee for Clinical Laboratory Standards document M27-A guidelines. Our data showed that there was a good correlation between the MICs obtained by the Etest and broth dilution methods. When only the MIC results at ± 2 dilutions for both methods were considered, the agreement rates were 90.4% for itraconazole, ketoconazole and amphotericin B and 84.6% for fluconazole of the C. albicans tested. In contrast, to the reference method, the Etest method classified as susceptible three fluconazole-resistant isolates and one itraconazole-resistant isolate, representing four very major errors. These results indicate that Etest could be considered useful for antifungal sensitivity evaluation of yeasts in clinical laboratories.

Per-unit royalty and fixed-fee licensing in a differentiated Stackelberg model

In the present paper we consider a differentiated Stackelberg model, when the leader firm engages in an R&D process that gives an endogenous cost-reducing innovation. The aim is to study the licensing of the cost-reduction by a per-unit royalty and a fixed-fee. We analyse the implications of these types of licensing contracts over the R&D effort, the profits of the firms, the consumer surplus and the social welfare. By using comparative static analysis, we conclude that the degree of the differentiation of the goods plays an important role in the results.

Head-Up Tilt Testing with Different Nitroglycerin Dosages: Experience in Elderly Patients with Unexplained Syncope

AIMS: Protocols using sublingual nitrates have been increasingly used to improve diagnostic accuracy of head-up tilt testing (HUT). Nevertheless, exaggerated responses to nitrates have been frequently described, particularly in elderly patients. The aim of this article is to evaluate, in an elderly population with unexplained syncope, whether the impact of sublingual nitroglycerin (NTG) used as a provocative agent is dose-dependent. METHODS AND RESULTS: One hundred and twenty consecutive elderly patients submitted to HUT using NTG after an asymptomatic drug-free phase were studied. Patients were divided into three groups according to the NTG dosage: 500, 375 and 250 microg. The test was considered positive when there was reproduction of symptoms with bradycardia and/or arterial hypotension. A gradual decrease in the blood pressure after NTG was considered an exaggerated response to nitrates. There were no differences in the clinical characteristics of the different subgroups. A positive test was obtained in 50% of the patients in each group. The rate of exaggerated responses was identical in all groups and ranged between 15 and 17%. CONCLUSION: In an elderly population with syncope of unknown origin submitted to HUT, the response to NTG is not dose-dependent, and no difference was found in the rate of exaggerated responses to nitrates with different NTG dosages.

Primary screening of blood donors by nat testing for HCV-RNA: development of an "in-house" method and results

An "in-house" RT-PCR method was developed that allows the simultaneous detection of the RNA of the Hepatitis C Virus (HCV) and an artificial RNA employed as an external control. Samples were analyzed in pools of 6-12 donations, each donation included in two pools, one horizontal and one vertical, permitting the immediate identification of a reactive donation, obviating the need for pool dismembering. The whole process took 6-8 hours per day and results were issued in parallel to serology. The method was shown to detect all six HCV genotypes and a sensitivity of 500 IU/mL was achieved (95% hit rate). Until July 2005, 139,678 donations were tested and 315 (0.23%) were found reactive for HCV-RNA. Except for five false-positives, all 310 presented the corresponding antibody as well, so the yield of NAT-only donations was zero, presenting a specificity of 99.83%. Detection of a window period donation, in the population studied, will probably demand testing of a larger number of donations. International experience is showing a rate of 1:200,000 - 1:500,000 of isolated HCV-RNA reactive donations.

In vitro susceptibility testing of dermatophytes isolated from pediatric cases in Nigeria against five antifungals

The antifungal activities of itraconazole, ketoconazole, fluconazole, terbinafine and griseofulvin were tested by broth microdilution methods against 71 isolates of dermatophytes isolated from Nigerian children. Most drugs were very active against all the dermatophytes and the MIC 90 ranged from 0.03 to 8.0 µg/mL. This appears to be the first documented data on the antifungal susceptibility testing of isolates of dermatophytes from Nigerian children.

Prognostic Value of a New Cardiopulmonary Exercise Testing Parameter in Chronic Heart Failure: Oxygen Uptake Efficiency at Peak Exercise - Comparison with Oxygen Uptake Efficiency Slope

INTRODUCTION: A growing body of evidence shows the prognostic value of oxygen uptake efficiency slope (OUES), a cardiopulmonary exercise test (CPET) parameter derived from the logarithmic relationship between O(2) consumption (VO(2)) and minute ventilation (VE) in patients with chronic heart failure (CHF). OBJECTIVE: To evaluate the prognostic value of a new CPET parameter - peak oxygen uptake efficiency (POUE) - and to compare it with OUES in patients with CHF. METHODS: We prospectively studied 206 consecutive patients with stable CHF due to dilated cardiomyopathy - 153 male, aged 53.3±13.0 years, 35.4% of ischemic etiology, left ventricular ejection fraction 27.7±8.0%, 81.1% in sinus rhythm, 97.1% receiving ACE-Is or ARBs, 78.2% beta-blockers and 60.2% spironolactone - who performed a first maximal symptom-limited treadmill CPET, using the modified Bruce protocol. In 33% of patients an cardioverter-defibrillator (ICD) or cardiac resynchronization therapy device (CRT-D) was implanted during follow-up. Peak VO(2), percentage of predicted peak VO(2), VE/VCO(2) slope, OUES and POUE were analyzed. OUES was calculated using the formula VO(2) (l/min) = OUES (log(10)VE) + b. POUE was calculated as pVO(2) (l/min) / log(10)peakVE (l/min). Correlation coefficients between the studied parameters were obtained. The prognosis of each variable adjusted for age was evaluated through Cox proportional hazard models and R2 percent (R2%) and V index (V6) were used as measures of the predictive accuracy of events of each of these variables. Receiver operating characteristic (ROC) curves from logistic regression models were used to determine the cut-offs for OUES and POUE. RESULTS: pVO(2): 20.5±5.9; percentage of predicted peak VO(2): 68.6±18.2; VE/VCO(2) slope: 30.6±8.3; OUES: 1.85±0.61; POUE: 0.88±0.27. During a mean follow-up of 33.1±14.8 months, 45 (21.8%) patients died, 10 (4.9%) underwent urgent heart transplantation and in three patients (1.5%) a left ventricular assist device was implanted. All variables proved to be independent predictors of this combined event; however, VE/VCO2 slope was most strongly associated with events (HR 11.14). In this population, POUE was associated with a higher risk of events than OUES (HR 9.61 vs. 7.01), and was also a better predictor of events (R2: 28.91 vs. 22.37). CONCLUSION: POUE was more strongly associated with death, urgent heart transplantation and implantation of a left ventricular assist device and proved to be a better predictor of events than OUES. These results suggest that this new parameter can increase the prognostic value of CPET in patients with CHF.

Comparison between E-test and CLSI broth microdilution method for antifungal susceptibility testing of Candida albicans oral isolates

Thirty Candida albicans isolated from oral candidosis patients and 30 C. albicans isolated from control individuals were studied. In vitro susceptibility tests were performed for amphotericin B, fluconazole, 5-flucytosine and itraconazole through the Clinical and Laboratorial Standards Institute (CLSI) reference method and E test system. The results obtained were analyzed and compared. MIC values were similar for the strains isolated from oral candidosis patients and control individuals. The agreement rate for the two methods was 66.67% for amphotericin B, 53.33% for fluconazole, 65% for flucytosine and 45% for itraconazole. According to our data, E test method could be an alternative to trial routine susceptibility testing due to its simplicity. However, it can not be considered a substitute for the CLSI reference method.

Desenvolvimento de Frameworks de Testes Automáticos de Software

O objetivo deste trabalho é o desenvolvimento de frameworks de testes automáticos de software. Este tipo de testes normalmente está associado ao modelo evolucionário e às metodologias ágeis de desenvolvimento de software, enquanto que os testes manuais estão relacionados com o modelo em cascata e as metodologias tradicionais. Como tal foi efetuado um estudo comparativo sobre os tipos de metodologias e de testes existentes, para decidir quais os que melhor se adequavam ao projeto e dar resposta à questão "Será que realmente compensa realizar testes (automáticos)?". Finalizado o estudo foram desenvolvidas duas frameworks, a primeira para a implementação de testes funcionais e unitários sem dependências a ser utilizada pelos estagiários curriculares da LabOrders, e a segunda para a implementação de testes unitários com dependências externas de base de dados e serviços, a ser utilizada pelos funcionários da empresa. Nas últimas duas décadas as metodologias ágeis de desenvolvimento de software não pararam de evoluir, no entanto as ferramentas de automação não conseguiram acompanhar este progresso. Muitas áreas não são abrangidas pelos testes e por isso alguns têm de ser feitos manualmente. Posto isto foram criadas várias funcionalidades inovadoras para aumentar a cobertura dos testes e tornar as frameworks o mais intuitivas possível, nomeadamente: 1. Download automático de ficheiros através do Internet Explorer 9 (e versões mais recentes). 2. Análise do conteúdo de ficheiros .pdf (através dos testes). 3. Obtenção de elementos web e respetivos atributos através de código jQuery utilizando a API WebDriver com PHP bindings. 4. Exibição de mensagens de erro personalizadas quando não é possível encontrar um determinado elemento. As frameworks implementadas estão também preparadas para a criação de outros testes (de carga, integração, regressão) que possam vir a ser necessários no futuro. Foram testadas em contexto de trabalho pelos colaboradores e clientes da empresa onde foi realizado o projeto de mestrado e os resultados permitiram concluir que a adoção de uma metodologia de desenvolvimento de software com testes automáticos pode aumentar a produtividade, reduzir as falhas e potenciar o cumprimento de orçamentos e prazos dos projetos das organizações.

Routine Transthoracic Echocardiography in a General Intensive Care Unit: an 18 Month Survey in 704 Patients

The authors analyzed 704 transthoracic echocardiographic (TTE) examinations, performed routinely to all admitted patients to a general 16-bed Intensive Care Unit (ICU) during an 18-month period. Data acquisition and prevalence of abnormalities of cardiac structures and function were assessed, as well as the new, previously unknown severe diagnoses. A TTE was performed within the first 24 h of admission on 704 consecutive patients, with a mean age of 61.5+/-17.5 years, ICU stay of 10.6+/-17.1 days, APACHE II 22.6+/-8.9, and SAPS II 52.7+/-20.4. In four patients, TTE could not be performed. Left ventricular (LV) dimensions were quantified in 689 (97.8%) patients, and LV function in 670 (95.2%) patients. Cardiac output (CO) was determined in 610 (86.7%), and mitral E/A in 399 (85.9% of patients in sinus rhythm). Echocardiographic abnormalities were detected in 234 (33%) patients, the most common being left atrial (LA) enlargement (n=163), and LV dysfunction (n=132). Patients with these alterations were older (66+/-16.5 vs 58.1+/-17.4, p<0.001), presented a higher APACHE II score (24.4+/-8.7 vs 21.1+/-8.9, p<0.001), and had a higher mortality rate (40.1% vs 25.4%, p<0.001). Severe, previously unknown echocardiographic diagnoses were detected in 53 (7.5%) patients; the most frequent condition was severe LV dysfunction. Through a multivariate logistic regression analysis, it was determined that mortality was affected by tricuspid regurgitation (p=0.016, CI 1.007-1.016) and ICU stay (p<0.001, CI 1-1.019). We conclude that TTE can detect most cardiac structures in a general ICU. One-third of the patients studied presented cardiac structural or functional alterations and 7.5% severe previously unknown diagnoses.

In vitro susceptibility testing of dermatophytes isolated in Goiania, Brazil, against five antifungal agents by broth microdilution method

The antifungal activities of fluconazole, itraconazole, ketoconazole, terbinafine and griseofulvin were tested by broth microdilution technique, against 60 dermatophytes isolated from nail or skin specimens from Goiania city patients, Brazil. In this study, the microtiter plates were incubated at 28 ºC allowing a reading of the minimal inhibitory concentration (MIC) after four days of incubation for Trichophyton mentagrophytes and five days for T. rubrum and Microsporum canis. Most of the dermatophytes had uniform patterns of susceptibility to the antifungal agents tested. Low MIC values as 0.03 µg/mL were found for 33.3%, 31.6% and 15% of isolates for itraconazole, ketoconazole and terbinafine, respectively.

Testing Nelder-Mead Based Repulsion Algorithms for Multiple Roots of Nonlinear Systems via a Two-Level Factorial Design of Experiments

This paper addresses the challenging task of computing multiple roots of a system of nonlinear equations. A repulsion algorithm that invokes the Nelder-Mead (N-M) local search method and uses a penalty-type merit function based on the error function, known as 'erf', is presented. In the N-M algorithm context, different strategies are proposed to enhance the quality of the solutions and improve the overall efficiency. The main goal of this paper is to use a two-level factorial design of experiments to analyze the statistical significance of the observed differences in selected performance criteria produced when testing different strategies in the N-M based repulsion algorithm. The main goal of this paper is to use a two-level factorial design of experiments to analyze the statistical significance of the observed differences in selected performance criteria produced when testing different strategies in the N-M based repulsion algorithm.