Predictive value of the acid fast smear for detection of Mycobacterium tuberculosis in respiratory specimens in a Reference Center of HIV/Aids in Rio de Janeiro, Brazil

In order to evaluate the predictive value of acid fast bacilii (AFB) smear for the diagnosis of Mycobacterium tuberculosis in respiratory specimens in a setting with a high prevalence of Aids and an unknown prevalence of nontuberculous mycobacteria (NTM), we retrospectively examined specimens cultured for mycobacteria between 1 September 1993 and 30 September 1994 and medical records of patients with positive culture in a General Hospital, Aids reference in Rio de Janeiro, Brazil. Seventy three per cent (1517/2077) of samples were respiratory specimens and mycobacteria were recovered from 20.6% (313/1517) of these. M. tuberculosis was identified in 94.2% (295/313) and NTM in 5.8% (18/313). The yield of positive AFB smear and of positive culture was 6.1% (93/1517) and 20.6% (313/1517), respectively. The positive predictive value (PPV) of AFB for M. tuberculosis was 98.4% in expectorated sputum and 96.4% in bronchoalveolar lavage. Forty four percent (130/295) of specimens with positive culture for M. tuberculosis and 66.7% (12/18) for NTM were from patients HIV positive. The conclusion was that in our study population, the PPV of AFB for M. tuberculosis in respiratory specimens was high and the prevalence of NTM was low despite the high prevalence of HIV positive.

Value of morphotyping for the characterization of Candida albicans clinical isolates

Until recently, morphotyping, a method evaluating fringe and surface characteristics of streak colonies grown on malt agar, has been recommended as a simple and unexpensive typing method for Candida albicans isolates. The discriminatory power and reproducibility of Hunter's modified scheme of Phongpaichit's morphotyping has been evaluated on 28 C. albicans isolates recovered from the oral cavity of asymptomatic human immunodeficiency virus-positive subjects, and compared to two molecular typing methods: randomly amplified polymorphic DNA (RAPD) fingerprinting, and contour clamped homogeneous electric field (CHEF) electrophoretic karyotyping. Morphological features of streak colonies allowed to distinguish 11 different morphotypes while RAPD fingerprinting yielded 25 different patterns and CHEF electrophoresis recognized 9 karyotypes. The discriminatory power calculated with the formula of Hunter and Gaston was 0.780 for morphotyping, 0.984 for RAPD fingerprinting, and 0.630 for karyotyping. Reproducibility was tested using 43 serial isolates from 15 subjects (2 to 6 isolates per subject) and by repeating the test after one year storage of the isolates. While genetic methods generally recognized a single type for all serial isolates from each of the subjects studied, morphotyping detected strain variations in five subjects in the absence of genetic confirmation. Poor reproducibility was demonstrated repeating morphotyping after one year storage of the isolates since differences in at least one character were detected in 92.9% of the strains.

The use of biodiversity as source of new chemical entities against defined molecular targets for treatment of malaria, tuberculosis, and T-cell mediated diseases: a review

The modern approach to the development of new chemical entities against complex diseases, especially the neglected endemic diseases such as tuberculosis and malaria, is based on the use of defined molecular targets. Among the advantages, this approach allows (i) the search and identification of lead compounds with defined molecular mechanisms against a defined target (e.g. enzymes from defined pathways), (ii) the analysis of a great number of compounds with a favorable cost/benefit ratio, (iii) the development even in the initial stages of compounds with selective toxicity (the fundamental principle of chemotherapy), (iv) the evaluation of plant extracts as well as of pure substances. The current use of such technology, unfortunately, is concentrated in developed countries, especially in the big pharma. This fact contributes in a significant way to hamper the development of innovative new compounds to treat neglected diseases. The large biodiversity within the territory of Brazil puts the country in a strategic position to develop the rational and sustained exploration of new metabolites of therapeutic value. The extension of the country covers a wide range of climates, soil types, and altitudes, providing a unique set of selective pressures for the adaptation of plant life in these scenarios. Chemical diversity is also driven by these forces, in an attempt to best fit the plant communities to the particular abiotic stresses, fauna, and microbes that co-exist with them. Certain areas of vegetation (Amazonian Forest, Atlantic Forest, Araucaria Forest, Cerrado-Brazilian Savanna, and Caatinga) are rich in species and types of environments to be used to search for natural compounds active against tuberculosis, malaria, and chronic-degenerative diseases. The present review describes some strategies to search for natural compounds, whose choice can be based on ethnobotanical and chemotaxonomical studies, and screen for their ability to bind to immobilized drug targets and to inhibit their activities. Molecular cloning, gene knockout, protein expression and purification, N-terminal sequencing, and mass spectrometry are the methods of choice to provide homogeneous drug targets for immobilization by optimized chemical reactions. Plant extract preparations, fractionation of promising plant extracts, propagation protocols and definition of in planta studies to maximize product yield of plant species producing active compounds have to be performed to provide a continuing supply of bioactive materials. Chemical characterization of natural compounds, determination of mode of action by kinetics and other spectroscopic methods (MS, X-ray, NMR), as well as in vitro and in vivo biological assays, chemical derivatization, and structure-activity relationships have to be carried out to provide a thorough knowledge on which to base the search for natural compounds or their derivatives with biological activity.

Beyond sepsis pathophysiology with cytokines: what is their value as biomarkers for disease severity?

Sepsis is a major challenge in medicine. It is a common and frequently fatal infectious condition. The incidence continues to increase, with unacceptably high mortality rates, despite the use of specific antibiotics, aggressive operative intervention, nutritional support, and anti-inflammatory therapies. Typically, septic patients exhibit a high degree of heterogeneity due to variables such as age, weight, gender, the presence of secondary disease, the state of the immune system, and the severity of the infection. We are at urgent need for biomarkers and reliable measurements that can be applied to risk stratification of septic patients and that would easily identify those patients at the highest risk of a poor outcome. Such markers would be of fundamental importance to decision making for early intervention therapy or for the design of septic clinical trials. In the present work, we will review current biomarkers for sepsis severity and especially the use of cytokines as biomarkers with important pathophysiological role.

Usefulness of PCR-based assays to assess drug efficacy in Chagas disease chemotherapy: value and limitations

One major goal of research on Chagas disease is the development of effective chemotherapy to eliminate the infection from individuals who have not yet developed cardiac and/or digestive disease manifestations. Cure evaluation is the more complex aspect of its treatment, often leading to diverse and controversial results. The absence of reliable methods or a diagnostic gold standard to assess etiologic treatment efficacy still constitutes a major challenge. In an effort to develop more sensitive tools, polymerase chain reaction (PCR)-based assays were introduced to detect low amounts of Trypanosoma cruzi DNA in blood samples from chagasic patients, thus improving the diagnosis and follow-up evaluation after chemotherapy. In this article, I review the main problems concerning drug efficacy and criteria used for cure estimation in treated chagasic patients, and the work conducted by different groups on developing PCR methodologies to monitor treatment outcome of congenital infections as well as recent and late chronic T. cruzi infections.

A new rapid colourimetric method for testing Mycobacterium tuberculosis susceptibility to isoniazid and rifampicin: a crystal violet decolourisation assay

The aim of this study was to investigate the performance of a new and accurate method for the detection of isoniazid (INH) and rifampicin (RIF) resistance among Mycobacterium tuberculosis isolates using a crystal violet decolourisation assay (CVDA). Fifty-five M. tuberculosis isolates obtained from culture stocks stored at -80ºC were tested. After bacterial inoculation, the samples were incubated at 37ºC for seven days and 100 µL of CV (25 mg/L stock solution) was then added to the control and sample tubes. The tubes were incubated for an additional 24-48 h. CV (blue/purple) was decolourised in the presence of bacterial growth; thus, if CV lost its colour in a sample containing a drug, the tested isolate was reported as resistant. The sensitivity, specificity, positive predictive value, negative predictive value and agreement for INH were 92.5%, 96.4%, 96.1%, 93.1% and 94.5%, respectively, and 88.8%, 100%, 100%, 94.8% and 96.3%, respectively, for RIF. The results were obtained within eight-nine days. This study shows that CVDA is an effective method to detect M. tuberculosis resistance to INH and RIF in developing countries. This method is rapid, simple and inexpensive. Nonetheless, further studies are necessary before routine laboratory implementation.

Bonafide, type-specific human papillomavirus persistence among HIV-positive pregnant women: predictive value for cytological abnormalities, a longitudinal cohort study

This study investigated the rate of human papillomavirus (HPV) persistence, associated risk factors, and predictors of cytological alteration outcomes in a cohort of human immunodeficiency virus-infected pregnant women over an 18-month period. HPV was typed through L1 gene sequencing in cervical smears collected during gestation and at 12 months after delivery. Outcomes were defined as nonpersistence (clearance of the HPV in the 2nd sample), re-infection (detection of different types of HPV in the 2 samples), and type-specific HPV persistence (the same HPV type found in both samples). An unfavourable cytological outcome was considered when the second exam showed progression to squamous intraepithelial lesion or high squamous intraepithelial lesion. Ninety patients were studied. HPV DNA persistence occurred in 50% of the cases composed of type-specific persistence (30%) or re-infection (20%). A low CD4+T-cell count at entry was a risk factor for type-specific, re-infection, or HPV DNA persistence. The odds ratio (OR) was almost three times higher in the type-specific group when compared with the re-infection group (OR = 2.8; 95% confidence interval: 0.43-22.79). Our findings show that bonafide (type-specific) HPV persistence is a stronger predictor for the development of cytological abnormalities, highlighting the need for HPV typing as opposed to HPV DNA testing in the clinical setting.

Aspectos biológicos de Zabrotes subfasciatus (Bohemann, 1833) (Coleoptera, Bruchidae) em Phaseolus vulgaris L., cv. Carioca (Fabaceae), sob condições de laboratório

Zabrotes subfasciatus is a serious pest of common beans, P. vulgaris L.. In Brazil there are several studies dealing with resistance of bean genotypes to this insect, while other studies have emphasized the utilization of oils and powders from plants to repel their attack. In this paper, fecundity, fertility, pattern of oviposition, life cycle and longevity were evaluated for a Brazilian stock from the Goiás State on P. vulgaris cv. Carioca, at 30ºC and 70% R.H. The mean fecundity was 38 eggs per female and 73% of viability. Egg laying showed an aggregated pattern. Males and females lived an average of 13 and 9 days, respectively. The total life cycle lasted for about 28 days.

Amostragem, diversidade e sazonalidade de Hemerobiidae (Neuroptera) em Coffea arabica L. cv. Obatã (Rubiaceae)

Este estudo teve por objetivo avaliar métodos de amostragem, abundância sazonal e diversidade da população de Hemerobiidae associada a cultivo de café Coffea arabica L. cv. Obatã em Cravinhos, São Paulo, Brasil. Para tanto foram realizadas amostragens semanais no período de maio de 2005 a abril de 2006. Os métodos de amostragem utilizados foram: rede de varredura e armadilhas de Möericke e luminosa. Foram coletados 491 exemplares de Hemerobiidae pertencentes a quatro gêneros: Nusalala (231 espécimes / 47,2% do total de hemerobiídeos coletados), Megalomus (110 / 22,5%), Hemerobius (104 / 21,3%) e Sympherobius (44 / 9%). A rede de varredura foi a mais eficiente para a captura de Hemerobiidae e a armadilha de Möericke foi o método de amostragem que apresentou os maiores valores de diversidade (H'= 0,56) e de equitabilidade (J= 0,93). Os hemerobiídeos estiveram presentes na área estudada durante o ano todo; as maiores freqüências foram registradas entre agosto e março (final do inverno, primavera e verão) e o maior pico populacional ocorreu em janeiro (na metade do verão). Megalomus apresentou correlação positiva e significativa (p< 0,05) com a precipitação pluviométrica e as temperaturas máxima e mínima; Nusalala com as temperaturas máxima e mínima e, Sympherobius apenas com a temperatura máxima.

Frações de zinco em solo arenoso e suas relações com disponibilidade para Cynodon spp cv. Tifton-85

Para avaliar um esquema de fracionamento de zinco em Argissolo arenoso e suas relações com a disponibilidade de Zn para Cynodon spp cv. Tifton-85, realizou-se um experimento em casa de vegetação, em esquema fatorial 5 x 2 x 2 (5 doses de Zn, 2 doses de calcário e 2 épocas de amostragem), em delineamento inteiramente casualizado, com três repetições. As doses de Zn foram de 0; 2,5; 5,0; 7,5 e 10 mg dm-3; metade dos vasos não recebeu calagem (V = 42 %) e metade recebeu a calagem com vistas em elevar o índice de saturação por bases a 70 %. As épocas de amostragem foram 30 e 150 dias após aplicação de Zn, respectivamente, antes do plantio e depois do 3º corte de Tifton-85. A aplicação de Zn resultou em aumento significativo do elemento nas frações: trocável, óxidos de Mn, matéria orgânica e óxidos de Fe. O Zn ligado aos óxidos de Mn aumentou significativamente com a calagem. Após 150 dias de experimentação, houve diminuição do Zn trocável, ligado aos óxidos de Mn, à matéria orgânica e aos óxidos de Fe, e aumento na fração residual. A distribuição de Zn nas frações do solo foi: residual > óxidos de Fe > óxidos de Mn > trocável > matéria orgânica. As relações entre as características estudadas mostraram que tanto o Zn-DTPA quanto o Zn trocável, ligado à M.O. e aos óxidos de Mn, foram eficientes para representar o Zn absorvido pela planta.

Eficiência da fertilização nitrogenada com uréia (15N) em Brachiaria brizantha cv. Marandu associada ao parcelamento de superfosfato simples e cloreto de potássio

Em pastagens, a avaliação da eficiência do processo de fertilização com N-uréia é importante em virtude das altas doses utilizadas. Vários trabalhos relatam que a aplicação de uréia associada com cloreto de potássio (KCl) e superfosfato simples (SFS) pode melhorar a sua eficiência. Na recuperação de pastagens degradadas ou em sistemas de exploração intensiva, as quantidades de KCl e SFS empregadas como fertilização corretiva são altas e aplicadas em novembro, no início da estação das águas. O parcelamento do KCl e SFS associado às coberturas nitrogenadas com uréia seria de fácil adoção sem custos adicionais. Com o objetivo de avaliar este manejo, foi desenvolvido um experimento em que se realizou o balanço anual da aplicação de 15N da uréia quando o SFS e, ou, o KCl foram parcelados, associados à aplicação da uréia em cinco coberturas, ou quando ambos foram aplicados de uma só vez, no mês de novembro. A maior produção de forragem ocorreu quando o SFS foi parcelado, seguido pelo parcelamento do KCl, da aplicação única de ambos e, finalmente, do parcelamento dos dois. A recuperação do N-uréia na parte aérea e no sistema solo-pastagem não diferiu com os tratamentos. Entretanto, a recuperação na coroa da planta e no sistema radicular diferiu entre os tratamentos e mostrou-se positivamente correlacionada com a produção de forragem. A recuperação de fertilizante na liteira foi favorecida pelo parcelamento do KCl. Apesar de não ter sido verificado aumento na recuperação total do sistema entre os tratamentos, o parcelamento do KCl e do SFS tem sido indicado por melhorar a recuperação do N-uréia em várias estruturas da planta, refletindo em maior produção da forrageira.

Atributos químicos e físicos de um argissolo cultivado com Panicum maximum Jacq. cv. IPR-86 Milênio, sob lotação rotacionada e adubado com nitrogênio

A avaliação dos atributos químicos e físicos do solo é necessária em estudos que exploram intensivamente os sistemas de pastejo. No presente trabalho, objetivou-se avaliar as variações nos atributos físicos e químicos de um Argissolo cultivado com capim-milênio, sob pastejo, no intervalo compreendido entre outubro/2001 e maio/2002, no quinto ano de aplicação sucessiva das seguintes doses de N: 0, 150, 300 e 450 kg ha-1. Após os cinco anos de aplicação de 450 kg ha-1 de N, o teor de MO aumentou em 5 g dm-3, o que resultou em aumento da CTC. No período de outubro/2001 a maio/2002 houve diminuição nos valores de pH e de V com a aplicação de N. A densidade do solo variou pouco com a adubação nitrogenada; a percentagem de microporos aumentou e a de macroporos diminuiu na camada de 0-5 cm com o aumento das doses de N, mas a porosidade total permaneceu constante; e a resistência à penetração, quando o solo apresentava 90 g kg-1 de água, não foi afetada pelas doses de N e não foi restritiva ao crescimento das plantas.

El espacio virtual de intercambio de información sobre recursos humanos en Ciencia y Tecnología de América Latina y el Caribe Del CV Lattes al CvLAC

Se describen los principales problemas de los diferentes sistemas de información para la gestión de la ciencia y tecnología de los países de América Latina y el Caribe y se identifican los retos y las alternativas para promover un mayor intercambio entre países del continente. La creación de un espacio de intercambio de información de los recursos humanos que participan en los sistemas de ciencia y tecnología, a través de una metodología común denominada el CvLAC, se constituye en una respuesta a los problemas identificados. El CvLAC parte de la experiencia brasileña del CvLattes. Se describen los objetivos y resultados esperados del Proyecto del CvLAC, ejecutado con la participación de Brasil, Colombia, Cuba, Chile, México y Venezuela. Se describen los desarrollos y alternativas a partir de los datos existentes en los países participantes en el proyecto, iniciando con Colombia y Chile, destacándose las infinitas posibilidades que las nuevas tecnologías de información digital y los nuevos desarrollos, logrados en este campo, hacen posible en términos de intercambio de datos.

Efeito da germinação de soja CV. BR-13 e Paraná sobre ácido fítico, fósforo total e inibidores de tripsina

Este trabalho foi conduzido para avaliar o efeito da germinação sobre os constituintes antinutricionais da soja, ácido fítico, P total e inibidores de tripsina, que interferem na utilização de proteínas e outras substâncias. A diminuição no aproveitamento de nutrientes resulta em inibição de crescimento, hipoglicemia ou danos a tecidos animais. Para os ensaios foram utilizadas as cultivares de soja (Glycine max (L.) Merrill ) BR-13 e Paraná. As sementes foram incubadas em câmara de germinação a 25°C e 100% de umidade, de 0 a 72 horas, com coleta de amostras em intervalos eqüidistantes de seis horas. A cultivar Paraná apresentou teores superiores de ácido fítico, P total e inibidores de tripsina, em relação à cultivar BR-13. As análises de variância e comparações entre médias indicaram que houve aumentos significativos (p < 0,05) nos teores de P total (8,5% e 3,6% nas cultivares BR-13 e Paraná, respectivamente) e nos níveis de inibidores de tripsina (22,05% e 17,10% nas cultivares BR-13 e Paraná, respectivamente), e reduções nos teores de ácido fítico (15,96% e 9,46% nas cultivares BR-13 e Paraná, respectivamente), em 72 horas de germinação.

Physiological changes during postharvest senescence of broccoli

The objective of this work was to determine the early physiological changes throughout shelf life of fresh broccoli (Brassica oleracea L. var. italica) cv. Piracicaba Precoce at 25ºC and relative humidity of 96% in the dark until complete senescence. Head inflorescences showed lack of turgidity and commercial value when weight loss reached up to 5%, coinciding with 48 hour after harvest. Chlorophyll content was stable until 24 hours after harvesting; afterwards, an intense degradation phase took place. At 72 hours, total head yellowing was observed when chlorophyll content dropped to 30% of its initial content. Peroxidase activity increased by 1.4 fold during the first six hours, dropping to its lowest level approximately 24 hours after harvesting. However, from this time on, an increment of activity was observed until 72 hours. At 24 hours after harvesting, respiration was reduced by 50%. At later stages of senescence, respiration of florets was stable, but in a lower level than that determined at harvest. Sharp reduction of starch and reducing sugars was observed within 24 hours after harvesting, followed by continuous period of decline in starch and non-reducing sugars.