Rapid detection of bovine coronavirus by a semi-nested RT-PCR

Bovine coronavirus (BCoV) is a member of the group 2 of the Coronavirus (Nidovirales: Coronaviridae) and the causative agent of enteritis in both calves and adult bovine, as well as respiratory disease in calves. The present study aimed to develop a semi-nested RT-PCR for the detection of BCoV based on representative up-to-date sequences of the nucleocapsid gene, a conserved region of coronavirus genome. Three primers were designed, the first round with a 463bp and the second (semi-nested) with a 306bp predicted fragment. The analytical sensitivity was determined by 10-fold serial dilutions of the BCoV Kakegawa strain (HA titre: 256) in DEPC treated ultra-pure water, in fetal bovine serum (FBS) and in a BCoV-free fecal suspension, when positive results were found up to the 10-2, 10-3 and 10-7 dilutions, respectively, which suggests that the total amount of RNA in the sample influence the precipitation of pellets by the method of extraction used. When fecal samples was used, a large quantity of total RNA serves as carrier of BCoV RNA, demonstrating a high analytical sensitivity and lack of possible substances inhibiting the PCR. The final semi-nested RT-PCR protocol was applied to 25 fecal samples from adult cows, previously tested by a nested RT-PCR RdRp used as a reference test, resulting in 20 and 17 positives for the first and second tests, respectively, and a substantial agreement was found by kappa statistics (0.694). The high sensitivity and specificity of the new proposed method and the fact that primers were designed based on current BCoV sequences give basis to a more accurate diagnosis of BCoV-caused diseases, as well as to further insights on protocols for the detection of other Coronavirus representatives of both Animal and Public Health importance.

The performance of semi-quantitative differential PCR is similar to that of real-time PCR for the detection of the MYCN gene in neuroblastomas

Amplification of the MYCN gene in neuroblastomas is a potent biological marker of highly aggressive tumors, which are invariably fatal unless sound clinical management is applied. To determine the usefulness of semi-quantitative differential PCR (SQ-PCR) for accurate quantification of MYCN gene copy number, we evaluated the analytical performance of this method by comparing the results obtained with it for 101 tumor samples of neuroblastoma to that obtained by absolute and relative real-time PCR. Similar results were obtained for 100 (99%) samples, no significant difference was detected between the median log10 MYCN copy number (1.53 by SQ-PCR versus 1.55 by absolute real-time PCR), and the results of the two assays correlated closely (r = 0.8, Pearson correlation; P < 0.001). In the comparison of SQ-PCR and relative real-time PCR, SQ-PCR versus relative real-time PCR concordant results were found in 100 (99%) samples, no significant difference was found in median log10 MYCN copy number (1.53 by SQ-PCR versus 1.27 by relative real-time PCR), and the results of the two assays correlated closely (r = 0.8, Pearson correlation; P < 0.001). These findings indicate that the performance of SQ-PCR was comparable to that of real-time PCR for the amplification and quantification of MYCN copy number. Thus, SQ-PCR can be reliably used as an alternative assay in laboratories without facilities for real-time PCR.

Study of the comprehension of the scientific method by members of a university health research laboratory

In Brazil, scientific research is carried out mainly at universities, where professors coordinate research projects with the active participation of undergraduate and graduate students. However, there is no formal program for the teaching/learning of the scientific method. The objective of the present study was to evaluate the comprehension of the scientific method by students of health sciences who participate in scientific projects in an academic research laboratory. An observational descriptive cross-sectional study was conducted using Edgar Morin complexity as theoretical reference. In a semi-structured interview, students were asked to solve an abstract logical puzzle - TanGram. The collected data were analyzed using the hermeneutic-dialectic analysis method proposed by Minayo and discussed in terms of the theoretical reference of complexity. The students’ concept of the scientific method is limited to participation in projects, stressing the execution of practical procedures as opposed to scientific thinking. The solving of the TanGram puzzle revealed that the students had difficulties in understanding questions and activities focused on subjects and their processes. Objective answers, even when dealing with personal issues, were also reflected on the students’ opinions about the characteristics of a successful researcher. Students’ difficulties concerning these issues may affect their scientific performance and result in poorly designed experiments. This is a preliminary study that should be extended to other centers of scientific research.

Multicomponent diffusion during Prato cheese ripening: mathematical modeling using the finite element method

The partial replacement of NaCl by KCl is a promising alternative to produce a cheese with lower sodium content since KCl does not change the final quality of the cheese product. In order to assure proper salt proportions, mathematical models are employed to control the product process and simulate the multicomponent diffusion during the reduced salt cheese ripening period. The generalized Fick's Second Law is widely accepted as the primary mass transfer model within solid foods. The Finite Element Method (FEM) was used to solve the system of differential equations formed. Therefore, a NaCl and KCl multicomponent diffusion was simulated using a 20% (w/w) static brine with 70% NaCl and 30% KCl during Prato cheese (a Brazilian semi-hard cheese) salting and ripening. The theoretical results were compared with experimental data, and indicated that the deviation was 4.43% for NaCl and 4.72% for KCl validating the proposed model for the production of good quality, reduced-sodium cheeses.

Biotechnological richness of the northeastern semi-arid region: antioxidant activity of casein hydrolysates from Moxotó goat milk (Capra hircus Linnaeus, 1758) obtained by papain action

This study aimed to identify antioxidant peptides from caprine casein hydrolysates by papain application using MALDI-TOF mass spectrometer, and a 2² full factorial design, with 4 axial points, in order to evaluate kinetic parameters (time and pH) effects on the degree of hydrolysis as well as the antioxidant activity of Moxotó goat milk casein peptides. Degree of hydrolysis was determined by total and soluble protein ratio in casein. Antioxidant activity was measured by ABTS method with 2, 2-cation-azinobis (3-ethylbenzothiazoline-6-sulfonic acid). TROLOX was used as standard. Peptide pattern and sequence of antioxidant amino acids were obtained using MALDI-TOF/MS. The highest degree of hydrolysis (28.5%) and antioxidant activity (2329.6 mmol.L TROLOX. mg- 1 peptide) were observed in the permeate. NENLL, NPWDQVK and LLYQEPVLGPV peptides, detected in the permeate, were pointed as the responsible for antioxidant activity, suggesting their potential application as food supplement and pharmaceutical products.

Propagação vegetativa de porta-enxertos de pereira por estacas semi-lenhosas

Pesquisou-se a viabilidade técnica da propagação vegetativa dos porta-enxertos de pereira 'Taiwan Nashi-C', 'Taiwan Mamenashi' (Pyrus calleryana Dcne.) e 'Seleção IAC-1' (Pyrus spp.), por estacas semi-lenhosas. O experimento foi conduzido em telado equipado com sistema de nebulização intermitente, pertencente ao Núcleo de Pesquisa e Desenvolvimento do Jardim Botânico, do Instituto Agronômico (IAC), em Campinas, Estado de São Paulo. Adotou-se o fatorial 3 x 4 (porta-enxertos x doses de ácido indolbutírico), com cinco repetições de 20 estacas, em delineamento inteiramente casualizado. Com os resultados obtidos aos 60 dias após a estaquia, conclui-se que: a) é tecnicamente possível a propagação dos porta-enxertos 'Taiwan Nashi-C', 'Taiwan Mamenashi' e 'Seleção IAC-1' por estacas semi-lenhosas; b) dentre os porta-enxertos pesquisados, 'Taiwan Nashi-C' apresenta a maior porcentagem de enraizamento, a menor porcentagem de estacas com calo, além do maior comprimento de raízes e número de raízes por estaca; c) é necessário o uso de ácido indolbutírico para o enraizamento de estacas semi-lenhosas desses porta-enxertos, sendo que, no conjunto das variáveis analisadas, as concentrações de 4.000 mg.L-1 e de 6.000 mg.L-1 proporcionam os maiores benefícios ao enraizamento adventício; d) as concentrações de ácido indolbutírico testadas não são fitotóxicas às estacas semi-lenhosas dos genótipos pesquisados.

Multiple centroid method to evaluate the adaptability of alfalfa genotypes

This study aimed to evaluate the efficiency of multiple centroids to study the adaptability of alfalfa genotypes (Medicago sativa L.). In this method, the genotypes are compared with ideotypes defined by the bissegmented regression model, according to the researcher's interest. Thus, genotype classification is carried out as determined by the objective of the researcher and the proposed recommendation strategy. Despite the great potential of the method, it needs to be evaluated under the biological context (with real data). In this context, we used data on the evaluation of dry matter production of 92 alfalfa cultivars, with 20 cuttings, from an experiment in randomized blocks with two repetitions carried out from November 2004 to June 2006. The multiple centroid method proved efficient for classifying alfalfa genotypes. Moreover, it showed no unambiguous indications and provided that ideotypes were defined according to the researcher's interest, facilitating data interpretation.

Survival analysis: a tool in the study of post-harvest diseases in peaches

Survival analysis is applied when the time until the occurrence of an event is of interest. Such data are routinely collected in plant diseases, although applications of the method are uncommon. The objective of this study was to use two studies on post-harvest diseases of peaches, considering two harvests together and the existence of random effect shared by fruits of a same tree, in order to describe the main techniques in survival analysis. The nonparametric Kaplan-Meier method, the log-rank test and the semi-parametric Cox's proportional hazards model were used to estimate the effect of cultivars and the number of days after full bloom on the survival to the brown rot symptom and the instantaneous risk of expressing it in two consecutive harvests. The joint analysis with baseline effect, varying between harvests, and the confirmation of the tree effect as a grouping factor with random effect were appropriate to interpret the phenomenon (disease) evaluated and can be important tools to replace or complement the conventional analysis, respecting the nature of the variable and the phenomenon.

Processo de trabalho e grupos semi-autônomos: a evolução da experiência sueca de Kalmar aos anos 90

Este trabalho visa a apresentar o desenvolvimento recente de novas formas de organização da produção e do trabalho na Suécia, em especial no que se refere ao setor automotivo (ônibus, caminhões e automóveis). Apresentam-se em detalhes as características da mais recente planta do grupo Volvo, denominada Uddevalla, projetada segundo o conceito de dock-assembly. É introduzida também uma discussão sucinta acerca de semelhanças e diferenças entre esse caso e outros como, por exemplo, o da planta da Toyota no Japão. Por fim, procurase reforçar a importância da consideração de soluções novas - e adaptadas - ao caso brasileiro, no que diz respeito a formas de organização do trabalho que contribuam para aumento da produtividade (em um sentido amplo) dos sistemas de produção.

An appraisal of the health planning method proposed by the Pan-American Health Organization for Latin America

A brief description of the main features of the health planning technique developed by the "Centro de Estudios del Desarrollo" (CENDES) in Venezuela, and proposed by the Pan-American Health Organization for use in Latin America, is presented. This presentation is followed by an appraisal of the planning method which includes comments both upon its positive aspects and upon its negative points. Comments are also made referring to other recent publications of the WHO/PAHO on health planning. In conclusion, the CENDES technique is considered a health planning method of great potential for use especially in underdeveloped areas, the success of its application depending upon the hability of the health planners to introduce the necessary modifications to adapt to the local circunstamces.

Prática do aleitamento materno em comunidades rurais do semi-árido baiano

Com o objetivo de conhecer p estabelecimento e duração do aleitamento materno total e exclusivo em comunidades rurais do Semi-Árido Baiano, Brasil, foram estudadas 226 crianças de O a 2 anos de idade, filhas de pequenos produtores rurais que vivem numa das regiões mais secas do nordeste brasileiro, de pobreza absoluta. Os dados referem-se a todas as crianças da família que à data do estudo tinham até 2 anos de idade. Os dados foram coletados por meio de formulários aplicados às mães ou responsáveis pelo menor, no período de 1988 a 1989. A tábua de vida foi o recurso estatístico utilizado para o cálculo da duração mediana do evento. Das 226 crianças que integraram o estudo, 91,6 iniciaram a amamentação ao peito. A duração mediana do aleitamento total foi de 90,28 dias e o aleitamento exclusivo apresentou duração mediana de 0,1 dia. Ainda que o estabelecimento da amamentação ao peito possa ser considerado alto, a duração mediana do aleitamento total foi extrememente curta, e o aleitamento exclusivo foi desprezível.

Hipovitaminose A em crianças de áreas rurais do semi-árido baiano

Objetivou-se avaliar a distribuição e a magnitude da deficiência de vitamina A e o consumo dietético de 161 crianças de 6 a 72 meses de idade, de áreas rurais do Município de Cansanção-Bahia, Brasil. Os níveis de retinol sérico foram medidos pelo método espectrofotométrico (Bessey-Lowry modificado por Araújo e Flores, 1978). A média do retinol sérico distribuiuse homogeneamente entre as diferentes faixas etárias. Níveis inadequados de retinol sérico (deficiente <10,0 µg/dl e baixos <20,0 µg/dl) foram detectados em 44,7% das crianças, caracterizando a deficiência como problema de saúde pública. Os níveis de retinol sérico não mostraram associação estatisticamente significante com sexo e idade das crianças, contudo as menores de 24 meses apresentaram prevalência mais alta de níveis inadequados. A principal fonte de vitamina A, disponível para essas crianças, é representada pelos carotenóides, em especial beta-caroteno. Foi observada maior diversificação no consumo dos alimentos de conteúdos moderado e baixo em vitamina A no grupo de 24 a 72 meses de idade, sem contudo assegurar níveis adequados de retinol sérico para este grupo etário.

Situação nutricional e alimentar de pré-escolares no semi-árido da Bahia (Brasil): I.Avaliação antropométrica

Foram estudados 754 pré-escolares de áreas urbanas de sete municípios do semi-árido da Bahia, Brasil. Os objetivos foram determinar a prevalência de déficit ponderal e estatural, indicativos de desnutrição atual e/ou pregressa e sua associação com a idade, sexo, renda em salário-mínimo (SM), escolaridade materna e adequação do consumo alimentar. Encontrou-se 22,9% de crianças com altura/idade abaixo de -2,0 DP (desnutrição pregressa), 19,1% com peso/idade e 3,6% com peso/altura abaixo de -2,0 DP (desnutrição atual). Em relação ao inquérito dietético somente 6,8% das crianças haviam consumido no dia anterior uma dieta que suprisse os requerimentos energéticos para sua faixa etária. Houve forte associação entre os indicadores A/l e P/l inadequados com renda familiar per capita (p=0,001 e p=0,000, respectivamente); crianças de famílias com renda per capita < 1/4 SM tinham duas vezes mais chance de estar desnutridas se comparadas com as do estrato de renda >1/2 SM. Em relação ao P/A como também P/l, os pré-escolares estudados no semi-árido apresentaram prevalências significativamente superiores às encontradas por pesquisa nacional realizada na mesma época (p=0,047 e p=0,000, respectivamente). Esses achados surpreendem, já que nas últimas décadas tem sido demonstrada no Brasil uma melhoria signifivativa na desnutrição e mortalidade infantil e parecem indicar que as crianças do semi-árido não lograram ainda alcançar os mesmos benefícios que o restante da população infantil brasileira.