Determination of pindolol enantiomers in human plasma and urine by simple liquid-liquid extraction and high-performance liquid chromatography

A simple method for the measurement of pindolol enantiomers by HPLC is presented. Alkalinized serum or urine is extracted with ethyl acetate and the residue remaining after evaporation of the organic layer is then derivatised with (S)-(-)-alpha-methylbenzyl isocyanate. The diastereoisomers of derivatised pindolol and metoprolol (internal standard) are separated by high-performance liquid chromatography (HPLC) using a C-18 silica column and detected using fluorescence (excitation lambda: 215 nm, emission lambda: 320 nm). The assay displays reproducible linearity for pindolol enantiomers with a correlation coefficient of r(2) greater than or equal to 0.998 over the concentration range 8-100 ng ml(-1) for plasma and 0.1-2.5 mu g ml(-1) for urine. The coefficient of variation for accuracy and precision of the quality control samples for both plasma and urine are consistently

Evaluation of an immunoassay (EMIT) for mycophenolic acid in plasma from renal transplant recipients compared with a high-performance liquid chromatography assay

Mycophenolic acid is an immunosuppressant administered as a bioavailable ester, mycophenolate mofetil. The pharmacokinetics of mycophenolic acid have been reported to be variable. Accurate measurement of concentrations of this drug could be important to adjust doses. The aim of this study was to compare the enzyme-multiplied immunoassay technique (EMIT [Dade Behring; San Jose, CA, U.S.A.]) for mycophenolic acid with a high-performance liquid chromatographic (HPLC) assay using samples collected from renal transplant recipients. The HPLC assay used solid phase extraction and a C18 stationary phase with ultraviolet (UV) detection (254 nm). The immunoassay required no manual sample preparation. Plasma samples (n = 102) from seven patients, collected at various times after a dose, were analyzed using both methods. Both assays fulfilled quality-control criteria. Higher concentrations were consistently measured in patient samples when using EMIT. The mean (+/- standard deviation [SD]) bias (EMIT-HPLC) was 1.88 +/- 0.86 mg/L. The differences in concentrations were higher in the middle of a dosage interval, suggesting that a metabolite might have been responsible for overestimation. Measurement of glucuronide concentrations by HPLC demonstrated only a weak correlation between assay differences and glucuronide concentrations. If the crossreacting substance is active, EMIT could provide a superior measure of immunosuppression; if inactive, further work is needed to improve antibody specificity. In conclusion, it was found that EMIT overestimates the concentration of mycophenolic acid in plasma samples from renal transplant recipients compared with HPLC analysis.

HER2 Testing in Breast Carcinoma Very Low Concordance Rate Between Reference and Local Laboratories in Brazil

Breast cancer accounts for approximately one quarter of all cancers in females. HER2 gene amplification or HER2 protein overexpression, detected in about 20% of breast carcinomas, predicts a more aggressive clinical course and determines eligibility for targeted therapy with trastuzumab. HER2 testing has become an essential part of the clinical evaluation of all breast carcinoma patients, and accurate HER2 results are critical in identifying patients who may be benefited from targeted therapy. This study investigated the concordance in the results of HER2 immunohistochemistry assays performed in 500 invasive breast carcinomas between a reference laboratory and 149 local laboratories from all geographic regions of Brazil. Our results showed an overall poor concordance (171 of 500 cases, 34.2%) regarding HER2 results between local and reference laboratories, which may be related to the low-volume load of HER2 assays, inexperience with HER2 scoring system, and/or technical issues related to immunohistochemistry in local laboratories. Standardization of HER2 testing with rigorous quality control measures by local laboratories is highly recommended to avoid erroneous treatment of breast cancer patients.

Study of the spatial distribution of the absorbed dose in blood volumes irradiated using a teletherapy unit

Blood irradiation can be performed using a dedicated blood irradiator or a teletherapy unit. A thermal device providing appropriate storage conditions during blood components irradiation with a teletherapy unit has been recently proposed. However, the most appropriated volume of the thermal device was not indicated. The goal of this study was to indicate the most appropriated blood volume for irradiation using a teletherapy unit in order to minimize both the dose heterogeneity in the volume and the blood irradiation time using these equipments. Theoretical and experimental methods were used to study the dose distribution in the blood volume irradiated using a linear accelerator and a cobalt-60 therapy machine. The calculation of absorbed doses in the middle plane of cylindrical acrylic volumes was accomplished by a treatment planning system. Experimentally, we also used cylindrical acrylic phantoms and thermoluminescent dosimeters to confirm the calculated doses. The data obtained were represented by isodose curves. We observed that an irradiation volume should have a height of 28 cm and a diameter of 28 cm and a height of 35 cm and a diameter of 35 cm, when the irradiation is to be performed by a linear accelerator and a cobalt-60 teletherapy unit, respectively. Calculated values of relative doses varied from 93% to 100% in the smaller volume, and from 66% to 100% in the largest one. A difference of 5.0%, approximately, was observed between calculated and experimental data. The size of these volumes permits the irradiation of blood bags in only one bath without compromising the homogeneity of the absorbed dose over the irradiated volume. Thus, these irradiation volumes can be recommend to minimize the irradiation time when a teletherapy unit is used to irradiate blood. (C) 2010 Elsevier Ltd. All rights reserved.

Venom-Sweet-Venom: N-Linked Glycosylation in Snake Venom Toxins

Protein glycosylation represents one of the most important post-translational events, and is a mean of diversifying a protein without recourse to the genome. The venoms produced by snakes contain an abundance of glycoproteins with N-linked carbohydrates. N-linked glycosylation can ensure the correct folding of important functional domains. Characterization of carbohydrates structures aids in development of human therapeutics by snake venom toxins.

Short communication: Identification of subclinical cow mastitis pathogens in milk by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

Subclinical mastitis is a common and easily disseminated disease in dairy herds. Its routine diagnosis via bacterial culture and biochemical identification is a difficult and time-consuming process. In this work, we show that matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) allows bacterial identification with high confidence and speed (1 d for bacterial growth and analysis). With the use of MALDI-TOF MS, 33 bacterial culture isolates from milk of different dairy cows from several farms were analyzed, and the results were compared with those obtained by classical biochemical methods. This proof-of-concept case demonstrates the reliability of MALDI-TOF MS bacterial identification, and its increased selectivity as illustrated by the additional identification of coagulase-negative Staphylococcus species and mixed bacterial cultures. Matrix-assisted laser desorption-ionization mass spectrometry considerably accelerates the diagnosis of mastitis pathogens, especially in cases of subclinical mastitis. More immediate and efficient animal management strategies for mastitis and milk quality control in the dairy industry can therefore be applied.

Telehealth in audiology: The need and potential to reach underserved communities

Permanent hearing loss is a leading global health care burden, with 1 in 10 people affected to a mild or greater degree. A shortage of trained healthcare professionals and associated infrastructure and resource limitations mean that hearing health services are unavailable to the majority of the world population. Utilizing information and communication technology in hearing health care, or tele-audiology, combined with automation offer unique opportunities for improved clinical care, widespread access to services, and more cost-effective and sustainable hearing health care. Tele-audiology demonstrates significant potential in areas such as education and training of hearing health care professionals, paraprofessionals, parents, and adults with hearing disorders; screening for auditory disorders; diagnosis of hearing loss; and intervention services. Global connectivity is rapidly growing with increasingly widespread distribution into underserved communities where audiological services may be facilitated through telehealth models. Although many questions related to aspects such as quality control, licensure, jurisdictional responsibility, certification and reimbursement still need to be addressed; no alternative strategy can currently offer the same potential reach for impacting the global burden of hearing loss in the near and foreseeable future.

Nucleation, growth and breakage phenomena in agitated wet granulation processes: a review

Wet agglomeration processes have traditionally been considered an empirical art, with great difficulties in predicting and explaining observed behaviour. Industry has faced a range of problems including large recycle ratios, poor product quality control, surging and even the total failure of scale up from laboratory to full scale production. However, in recent years there has been a rapid advancement in our understanding of the fundamental processes that control granulation behaviour and product properties. This review critically evaluates the current understanding of the three key areas of wet granulation processes: wetting and nucleation, consolidation and growth, and breakage and attrition. Particular emphasis is placed on the fact that there now exist theoretical models which predict or explain the majority of experimentally observed behaviour. Provided that the correct material properties and operating parameters are known, it is now possible to make useful predictions about how a material will granulate. The challenge that now faces us is to transfer these theoretical developments into industrial practice. Standard, reliable methods need to be developed to measure the formulation properties that control granulation behaviour, such as contact angle and dynamic yield strength. There also needs to be a better understanding of the flow patterns, mixing behaviour and impact velocities in different types of granulation equipment. (C) 2001 Elsevier Science B.V. All rights reserved.

Rapid diagnosis in pediatric infectious diseases: The past, the present and the future

The focus of rapid diagnosis of infectious diseases of children in the last decade has shifted from variations of the conventional laboratory techniques of antigen detection, microscopy and culture to that of molecular diagnosis of infectious agents. Pediatricians will need to be able to interpret the use, limitations and results of molecular diagnostic techniques as they are increasingly integrated into routine clinical microbiology laboratory protocols. PCR is the best known and most successfully implemented diagnostic molecular technology to date. It can detect specific infectious agents and determine their virulence and antimicrobial genotypes with greater speed, sensitivity and specificity than conventional microbiology methods. Inherent technical limitations of PCR are present, although they are reduced in laboratories that follow suitable validation and quality control procedures. Variations of PCR together with advances in nucleic acid amplification technology have broadened its diagnostic capabilities in clinical infectious disease to now rival and even surpass traditional methods in some situations. Automation of all components of PCR is now possible. The completion of the genome sequencing projects for significant microbial pathogens, in combination with PCR and DNA chip technology, will revolutionize the diagnosis and management of infectious diseases.

Quantification and stability of everolimus (SDZ RAD) in human blood by high-performance liquid chromatography-electrospray tandem mass spectrometry

We report here a validated method for the quantification of a new immunosuppressant drug, everolimus (SDZ RAD), using HPLC-tandem mass spectrometry. Whole blood samples (500 mul) were prepared by protein precipitation, followed by C-18 solid-phase extraction. Mass spectrometric detection was by selected reaction monitoring with an electrospray interface operating in positive ionization mode. The assay was linear from 0.5 to 100 mug/l (r(2) > 0.996, n = 9). The analytical recovery and inter-day imprecision, determined using whole blood quality control samples (n = 5) at 0.5, 1.2, 20.0, and 75.0 mug/l, was 100.3-105.4% and less than or equal to7.6%, respectively. The assay had a mean relative recovery of 94.8 +/- 3.8%. Extracted samples were stable for up to 24 h. Fortified everolimus blood samples were stable at -80 degreesC for at least 8 months and everolimus was found to be stable in blood when taken through at least three freeze-thaw cycles. The reported method provides accurate, precise and specific measurement of everolimus in blood over a wide analytical range and is currently supporting phase 11 and III clinical trials. (C) 2002 Elsevier Science B.V. All rights reserved.

Prolonged retention after aggregation into secretory granules of human R183H-growth hormone (GH), a mutant that causes autosomal dominant GH deficiency type II

Human R183H-GH causes autosomal dominant GH deficiency type II. Because we show here that the mutant hormone is fully bioactive, we have sought to locate an impairment in its progress through the secretory pathway as assessed by pulse chase experiments. Newly synthesized wild-type and R183H-GH were stable when expressed transiently in AtT20 cells, and both formed equivalent amounts of Lubrol-insoluble aggregates within 40 min after synthesis. There was no evidence for intermolecular disulfide bond formation in aggregates of wild-type hormone or the R183H mutant. Both wildtype and R183H-GH were packaged into secretory granules, assessed by the ability of 1 mm BaCl2 to stimulate release and by immunocytochemistry. The mutant differed from wildtype hormone in its retention in the cells after packaging into secretory granules; 50% more R183H-GH than wild-type aggregates were retained in AtT20 cells 120 min after synthesis, and stimulated release of R183H-GH or a mixture of R183H-GH and wild-type that had been retained in the cell was reduced. The longer retention of R183H-GH aggregates indicates that a single point mutation in a protein contained in secretory granules affects the rate of secretory granule release.

Controle estatístico de processo aplicado no monitoramento da absorção de água por carcaças de frango durante a etapa de pré-resfriamento

A absorção de água por carcaças de frango na etapa de pré-resfriamento da linha abate representa uma característica de qualidade importante relacionada ao rendimento do produto final. Uma forma de manter o padrão de qualidade de um produto é garantir que as etapas do processo sejam estáveis e replicáveis. Ao empregar o Controle Estatístico de Processo (CEP) é possível obter estabilidade e melhorias nos processos, por meio da redução da variabilidade. Neste contexto, o objetivo deste trabalho foi a aplicação de gráficos de controle, análise de correlação, estatística descritiva, testes de hipóteses e regressão linear múltipla na linha de abate de um abatedouro-frigorífico de aves para monitorar a variabilidade da absorção de água pelas carcaças de frango após a etapa de pré-resfriamento. Como resultado, verificou-se que o teor de absorção de água das carcaças de frango apresentou elevada variabilidade, sendo que 10% (8/80) das carcaças apresentaram absorção de água superior ao limite de 8% definido pela legislação brasileira. Do total de 16 variáveis de entrada analisadas, as mais impactantes no teor de absorção de água foram o “tempo de retenção da carcaça no pré-chiller” e o “tempo de espera da carcaça após a etapa de gotejamento”. Entretanto, o modelo de regressão obtido apresentou baixa correlação (R²=0,16) que foi associada à elevada variabilidade da variável-resposta. Os resultados da estatística descritiva demonstraram que as variáveis de entrada também apresentaram elevada variabilidade, com coeficiente de variação entre 7,95 e 63,5%. Verificou-se, pela análise dos gráficos de controle de medida individual e da amplitude móvel, que 15 das 16 variáveis de entrada se apresentaram fora de controle estatístico assim como a variável-resposta. Baseado no fluxograma e na descrição das etapas da linha de abate, previamente realizados, atribuiu-se à falta de padronização na condução das etapas e de procedimentos para o controle de qualidade das operações na linha de abate como fatores relevantes que poderiam estar associados à presença de causas especiais no processo. Concluiu-se que para reduzir a elevada variabilidade das variáveis e eliminar as causas especiais presentes são necessários ajustes operacionais para, dessa forma, obter um processo mais estável e mais uniforme garantindo o padrão de qualidade das carcaças de frango em relação ao teor de absorção de água.

Em busca de contribuições Schumpeterianas à Teoria da Regulação : as telecomunicações brasileiras numa perspectiva evolucionária

A partir da década de 1970, a literatura que trata daqueles setores de infraestrutura, cujo funcionamento requer um acompanhamento específico do Estado, vale dizer através da regulação, absorveu grande parte dos conceitos fundamentais desenvolvidos pelas escolas institucionalistas, notadamente pela Nova Economia Institucional (NEI). Entretanto, apesar dos avanços alcançados, esta literatura ainda carece de alguns desenvolvimentos teóricos e estudos empíricos que contribuam de forma decisiva. Tais questões estão ligadas, por exemplo, à evolução dos cenários e às condições econômicas que são alteradas a partir de mudanças significativas nas condições tecnológicas e que acabam por alterar significativamente aquela estrutura industrial para qual o modelo regulatório foi desenhado. Nesse sentido, as contribuições da Economia da Regulação e da NEI carecem de elementos evolucionários, notadamente schumpeterianos. O principal objetivo da dissertação é investigar como alguns elementos de inspiração schumpeteriana podem em algum grau ser aplicados à economia da regulação e neste sentido contribuir com proposições naqueles temas que essa escola fincou sua abordagem característica: o progresso tecnológico e o comportamento estratégico inovativo das empresas. Nesse sentido, o problema de pesquisa desse trabalho pode ser assim colocado: como pode ser entendido o papel do progresso tecnológico e da inovação nas teorias da regulação? A hipótese assumida neste trabalho é que não se pode desconsiderar a tecnologia e suas diversas implicações para a análise dos setores regulados, constatação essa cada vez mais clara, dadas as recentes transformações dessas indústrias: uma das principais características dos setores regulados, a partir da década de 1990, é o surgimento de novos marcos regulatórios e uma alteração progressiva dos modelos organizacionais que predominavam até então e um movimento de adaptação à margem da regulação vigente. Os resultados empíricos mostram que as transformações tecnológicas e institucionais ocorridas a partir de 1995 na indústria brasileira de telecomunicações alteraram significativamente a estrutura industrial e a dinâmica concorrencial do setor, colocando ao desenho regulatório – da Anatel – novos e complexos desafios, sobretudo, o de se adaptar à margem da dinamicidade característica das telecomunicações. Neste novo contexto, as funções regulatórias são mais complexas, o que exige um duplo processo de aprendizagem: quanto às estruturas de mercado e quanto ao comportamento estratégico das empresas.

Indoor school environment: easy and low cost to assess inorganic pollutants

Total particulate matter (TPM) was passively collected inside two classrooms of each of five elementary schools in Lisbon, Portugal. TPM was collected in polycarbonate filters with a 47 mm diameter, placed inside of uncovered plastic petri dishes. The sampling period was from 19 May to 22 June 2009 (35 days exposure) and the collected TPM masses varied between 0.2 mg and 0.8 mg. The major elements were Ca, Fe, Na, K, and Zn at μg level, while others were at ng level. Pearson′s correlation coefficients above 0.75 (a high degree of correlation) were found between several elements. Soil-related, traffic soil re-suspension and anthropogenic emission sources could be identified. Blackboard chalk was also identified through Ca large presence. Some of the determined chemical elements are potential carcinogenic. Quality control of the results showed good agreement as confirmed by the application of u-score test.

Avaliação da qualidade dos dados populacionais e cobertura dos registros de óbitos para as regiões brasileiras

OBJETIVO: Avaliar a qualidade dos dados populacionais e a cobertura dos registros de óbitos para as Unidades da Federação Brasileira (UF), por sexo, no ano de 1990. MÉTODOS: Os dados populacionais extraídos dos Censos Demográficos foram avaliados através de métodos demográficos clássicos. Os registros de óbitos, extraídos da Fundação Instituto Brasileiro de Geografia e Estatística e do Ministério da Saúde, foram avaliados através de três técnicas de mensuração do sub-registro de óbitos. RESULTADOS: A precisão das declarações das idades das UF, em sua maioria, passou do status de "pouco preciso" ou "moderado" para "preciso" na década de 80. A cobertura dos óbitos, em 1990, foi classificada como "boa" ou "satisfatória" para todos os Estados do Sul, Sudeste, Centro-Oeste e para os do Nordeste, abaixo do Rio Grande do Norte. O restante dos estados foram classificados como "regular" ou "insatisfatório". CONCLUSÕES: Houve uma melhora importante na qualidade das declarações da população recenseada e aumento da cobertura de óbitos e que, para muitas UF, é possível gerar confiáveis indicadores de mortalidade.