Time course of in vitro maturation of intra-erythrocytic malaria parasite: a comparison between Plasmodium falciparum and Plasmodium knowlesi

The schizont maturation assay for in vitro drug sensitivity tests has been a standard method employed in the global baseline assessment and monitoring of drug response in Plasmodium falciparum. This test is limited in its application to synchronous plasmodial infections because it evaluates the effect of drug on the maturation of parasite especially from ring to schizont stage and therefore synchronized P. falciparum cultures are required. On the other hand, P. knowlesi, a simian malaria parasite has a unique 24-h periodicity and maintains high natural synchronicity in monkeys. The present report presents the results of a comparative study on the course of in vitro maturation of sorbitol synchronized P. falciparum and naturally synchronous P. knowlesi. Ring stage parasites were incubated in RPMI medium supplemented with 10-15% pooled homologous serum in flat-bottomed 96-well micro plates using a candle jar at 37°C. The results suggest that the ideal time for harvesting the micro-assay plates for in vitro drug sensitivity test for sorbitol-synchronized P. falciparum and naturally synchronous P. knowlesi are from 26 to 30 h and from 22 to 25 h, respectively. The advantages of using P. knowlesi in chemotherapeutic studies are discussed.

Efeito da sacarose e sorbitol na conservação in vitro de Passiflora giberti N. E. Brown

Este trabalho teve como objetivo estudar o efeito da sacarose e do sorbitol na conservação in vitro de um acesso de Passiflora giberti N. E. Brown. Para isso, foi instalado um experimento no delineamento inteiramente casualizado, em que foi comparado o tratamento-testemunha (MS padrão) com o meio MS suplementado com três concentrações de sacarose (0; 15 e 30 g L-1) em combinação com três concentrações de sorbitol (10; 20 e 40 g L-1). As avaliações foram realizadas aos 30; 60; 90 e 120 dias de incubação, observando-se o comprimento das brotações (cm), número de raízes, número e coloração das folhas. Os resultados mostram ser possível conservar sob crescimento lento, por quatro meses, microplantas de maracujazeiro em meio de cultura MS suplementado com 10 ou 20 g L-1 de sorbitol, na ausência de sacarose, e mantidas sob condições de fotoperíodo de 16 h (22 µE m-2s-1) e temperatura de 27 ± 1 ºC. A sacarose promoveu maior desenvolvimento de microplantas. A rizogênese é afetada pelo sorbitol na concentração de 40 g L-1 e pela ausência de sacarose no meio de cultura.

Synchrony is more than overlap: measuring phenological synchronization considering time length and intensity

The degree of flowering and fruiting synchronization is believed to have ecological and evolutionary relevance at several scales. Here we discuss some measures that have been used to estimate synchrony and propose an index that incorporates both the entire length of an individual phenophase and variation in the number of flowers or fruits over that time period. This new index describes more accurately the phenological synchrony among individuals and populations.

Synchronization of ovulation in crossbred dairy heifers using gonadotrophin-releasing hormone agonist, prostaglandin F2a and human chorionic gonadotrophin or estradiol benzoate

Girolando (Gir x Holstein) is a very common dairy breed in Brazil because it combines the rusticity of Gir (Bos indicus) with the high milk yield of Holstein (Bos taurus). The ovarian follicular dynamics and hormonal treatments for synchronization of ovulation and timed artificial insemination were studied in Girolando heifers. The injection of a gonadotrophin-releasing hormone (GnRH) agonist was followed 6 or 7 days (d) later by prostaglandin F2a (PGF2a). Twenty-four hours after PGF2a injection either human chorionic gonadotropin (hCG, GPh-d6 and GPh-d7 groups) or estradiol benzoate (EB, GPE-d6 and GPE-d7 groups) was administered to synchronize ovulation and consequently allow timed artificial insemination (AI) 24 and 30 h after hCG and EB injection, respectively. Follicular dynamics in Girolando heifers was characterized by the predominance of three follicular waves (71.4%) with sizes of dominant follicles (10-13 mm) and corpus luteum (approximately 20 mm) similar to those for Bos indicus cattle. In the GnRH-PGF-hCG protocol, hCG administration induced earlier ovulation (67.4 h, P<0.01) compared to the control group (GnRH-PGF) and a better synchronization of ovulation, since most of it occurred within a period of 12 to 17 h. Pregnancy rate after timed AI was 42.8 (3/7, GPh-d6) to 50% (7/14, GPh-d7). In contrast, estradiol benzoate (GnRH-PGF-EB protocol) synchronized ovulation of only 5 of 11 heifers from the GPE-d7 group and of none (0/7) from the GPE-d6 group, which led to low pregnancy rates after timed AI (27.3 and 0%, respectively). However, since a small number of Girolando heifers was used to determine pregnancy rates in the present study, pregnancy rates should be confirmed with a larger number of animals.

Cell cycle synchronization and BrdU incorporation as a tool to study the possible selective elimination of ErbB1 gene in the micronuclei in A549 cells

Lung cancer often exhibits molecular changes, such as the overexpression of the ErbB1 gene that encodes epidermal growth factor receptor (EGFR). ErbB1 amplification and mutation are associated with tumor aggressiveness and low response to therapy. The aim of the present study was to design a schedule to synchronize the cell cycle of A549 cell line (a non-small cell lung cancer) and to analyze the possible association between the micronuclei (MNs) and the extrusion of ErbB1 gene extra-copies. After double blocking, by the process of fetal bovine serum deprivation and vincristine treatment, MNs formation was monitored with 5-bromo-2-deoxyuridine (BrdU) incorporation, which is an S-phase marker. Statistical analyses allowed us to infer that MNs may arise both in mitosis as well as in interphase. The MNs were able to replicate their DNA and this process seemed to be non-synchronous with the main cell nuclei. The presence of ErbB1 gene in the MNs was evaluated by fluorescent in situ hybridization (FISH). ErbB1 sequences were detected in the MNs, but a relation between the MNs formation and extrusion of amplified ErbB1could not be established. The present study sought to elucidate the meaning of MNs formation and its association with the elimination of oncogenes or other amplified sequences from the tumor cells.

Determination of structural and mechanical properties, diffractometry, and thermal analysis of chitosan and hydroxypropylmethylcellulose (HPMC) films plasticized with sorbitol

In this work, the structural, mechanical, diffractometric, and thermal parameters of chitosan-hydroxypropylmethylcellulose (HPMC) films plasticized with sorbitol were studied. Solutions of HPMC (2% w/v) in water and chitosan (2% w/v) in 2% acetic acid solution were prepared. The concentration of sorbitol used was 10% (w/w) to both polymers. This solutions were mixed at different proportions (100/0; 70/30; 50/50; 30/70, and 0/100) of chitosan and HPMC, respectively, and 20 mL was cast in Petri dishes for further analysis of dried films. The miscibility of polymers was assessed by X-ray diffraction, scanning electronic microscopy (SEM), differential scanning calorimetry (DSC), and thermal gravimetric analysis (TGA). The results obtained indicate that the films are not fully miscible at a dry state despite the weak hydrogen bonding between the polymer functional groups.

Price synchronization in retailing: some empirical evidence

The paper investigates the synchronization of price changes in the context of retail tire dealers in São Paulo-Brazil and selected items in supermarkets for cleaning supplies and food in Rio de Janeiro-Brazil. Results indicate similar and non-negligible synchronization for different brands, although magnitudes are distant from a perfect synchronization pattern. We find interesting patterns in inter-firm competition, with similar magnitudes across different tire types. Intra-chain synchronization is substantial, indicating that a common price adjustment policy tends to be sustained for each chain across different products.

Excipientes de medicamentos e as informações da bula

OBJETIVO: Avaliar a presença de conservantes, corantes, adoçantes e aromatizantes em 73 apresentações farmacêuticas de 35 medicamentos para uso oral, e as informações da bula sobre excipientes. MÉTODOS: Selecionamos 35 medicamentos, de venda livre ou sob prescrição médica, comercializados no Brasil. A amostra incluiu: analgésicos/antitérmicos, antimicrobianos, mucolíticos, antitussígenos, descongestionantes, anti-histamínicos, broncodilatadores, corticosteróides, antiinflamatórios e suplementos vitamínicos. Foram analisadas 73 apresentações desses fármacos, anotando-se as informações da bula sobre conservantes, corantes, adoçantes e aromatizantes. RESULTADOS: A bula de um medicamento (1,3%) não mencionava os ingredientes inativos. Os conservantes mais encontrados nos medicamentos foram metilparabeno e propilparabeno (43% e 35,6% respectivamente). Os adoçantes mais usados foram: sacarose (açúcar) (53,4%), sacarina sódica (38,3%) e sorbitol (36,9%). Vinte e um produtos (28,7%) continham dois adoçantes. Predominaram os medicamentos sem corante (43,8%), seguidos pelos coloridos por amarelo crepúsculo (amarelo FD&C no. 6) (15%). Cinco produtos (6,8%) continham mais de um corante. A tartrazina (amarelo FD&C no. 5) foi encontrada em sete formulações (9,5%). Os aromatizantes mais usados foram os de frutas (83%). Constatamos a freqüente omissão das bulas sobre o teor exato de açúcar dos produtos (77%). Duas das quatro bulas de medicamentos contendo aspartame não mencionavam as precauções no uso por fenilcetonúricos. CONCLUSÕES: A omissão e a imprecisão das informações da bula sobre os excipientes farmacêuticos expõem os indivíduos suscetíveis ao risco de reações adversas dos conservantes e corantes. Também podem ocorrer complicações do uso inadvertido de medicamentos contendo açúcar pelos pacientes diabéticos, ou de fármacos adoçados com aspartame pelos fenilcetonúricos.

Análise físico-química e sensorial de doce de leite produzido sem adição de sacarose

O objetivo foi avaliar doces de leite produzidos sem adição de açúcar. Realizaram-se três formulações: doce com adição de açúcar (A), com edulcorante sucralose (B) e doce com ciclamato, sacarina e sorbitol (C). Nos doces B e C utilizou-se a carragena. Realizaram-se análises físico-quimicas, microbiológicas, exigidas pela legislação vigente, e análise sensorial. Nas análises de acidez não houve diferença significativa entre as amostras. O teor de gordura, a atividade de água e textura das amostras não diferiram entre os doces B e C, e esses diferiram estatisticamente do A. Nas análises de pH houve diferença significativa entre os doces. O teor de sólidos solúveis do doce A apresentou 63 °Brix, e os doces B e C, 26 °Brix. Na aceitação, observou-se diferença significativa para todos os atributos avaliados. Para sabor, o doce A foi mais aceito que os doces B e C, e o doce B mais que o C. Para atributo de textura sensorial e impressão global, o doce A foi mais aceito que os doces B e C, e esses apresentaram a mesma aceitação. Na análise de textura instrumental, os doces B e C não obtiveram a mesma força de resistência à penetração que o doce A. Todas as análises microbiológicas dos doces A, B e C apresentaram-se dentro dos padrões exigidos pela legislação vigente. Doce de leite com sucralose (B) mostrou-se uma boa alternativa para substituição do doce de leite com sacarose (A), uma vez que obteve boa aceitação para o atributo sabor.

Norfloxacin monodose use in patients with cholera in Salta, Argentina

The use ofmonodose (800 mg) per os of Norfloxacin was evaluated in 32 patients with cholera at Salvador Mazza's Hospital, Salta, Argentina. It was considered the celerity in negativization of stool culture (100% of cases: 12 hours post administration), its efficiency along time (24/24 controlled patients were negative at 10th day) and MIC of isolated strains (100% of strains were sensitive: range 0.008 to 0.016 mug/ml). It was included oral administration of sorbitol 70% in peanut oil in order to study patients at 10th day's control. This method could be an alternative one in the study of asymptomatic carrier. Norfloxacin monodose shows good performance in early negativization of stool culture and it was also effective along the whole observation period, suggesting it could prevent carriage.

The influence of carbohydrates in the interaction of Paracoccidioides brasiliensis with CCL-6 cells in vitro

INTRODUCTION: Little is known about the early events in the interaction between Paracoccidioides brasiliensis and its host. To understand the effect of carbohydrates in the interaction between the fungus and epithelial cell in culture, we analyzed the influence of different carbohydrate solutions on the adhesion of P. brasiliensis yeast cells to CCL-6 cells in culture. METHODS: Fungal cells were cultivated with the epithelial cell line, and different concentrations of D-fucose, N-acetyl-glucosamine, D-mannose, D-glucosamine, D-galactosamine, sorbitol and fructose were added at the beginning of the experiment. Six hours after the treatment, the cells were fixed and observed by light microscopy. The number of P. brasiliensis cells that were adhered to the CCL-6 monolayer was estimated. RESULTS: The number of adhesion events was diminished following treatments with D-fucose, N-acetyl-glucosamine, D-mannose, D-glucosamine and D-galactosamine as compared to the untreated controls. Sorbitol and fructose-treated cells had the same adhesion behavior as the observed in the control. P. brasiliensis propagules were treated with fluorescent lectins. The FITC-labeled lectins WGA and Con-A bound to P. brasiliensis yeast cells, while SBA and PNA did not. CONCLUSIONS: The perceptual of adhesion between P. brasiliensis and CCL-6 cells decreased with the use of D-mannose, N-acetyl-glucosamine and D-glucosamine. The assay using FITC-labeled lectins suggests the presence of N-acetyl-glucosamine, α-mannose and α-glucose on the P. brasiliensis cell surface. An enhanced knowledge of the mediators of adhesion on P. brasiliensis could be useful in the future for the development of more efficient and less harmful methods for disease treatment and control.

Dessincronia ventricular e aumento dos níveis de BNP na estimulação apical do ventrículo direito

FUNDAMENTO: A estimulação apical crônica do ventrículo direito pode ocasionar dessincronia ventricular e, secundariamente, alterações neuro-humorais e aumento da morbimortalidade cardíaca. OBJETIVO: Pesquisar dessincronia ventricular e seus efeitos sobre os níveis de BNP em pacientes com marca-passo estimulados cronicamente no ápice do ventrículo direito (VD). MÉTODOS: Estudo transversal com 85 pacientes com marca-passo uni ou bicameral, em classe funcional I e II da NYHA e fração de ejeção do ventrículo esquerdo (FEVE) > 35%. A avaliação de dessincronia foi realizada utilizando-se várias técnicas ecocardiográficas, incluindo o Tissue Synchronization Imaging (TSI), com análise dos 12 segmentos. O BNP foi dosado junto com o ecocardiograma, porém com o examinador cego. RESULTADOS: Quarenta e seis mulheres e 39 homens, com idade de 58 ± 12 anos, chagásicos (56%) e hipertensos controlados (62%), foram incluídos. A fração de ejeção do VE foi 52 ± 8% e a duração média do QRS de 139 ms (120-180 ms). O BNP mostrou-se alterado em 36,5% da amostra (ponto de corte de 60 pg/ml). Na análise multivariada de regressão linear, o BNP correlacionou-se com a idade (p = 0,024), FEVE (p < 0,0001) e tempo pré-ejetivo do VE (p = 0,009), que é índice de dessincronia intraventricular. CONCLUSÃO: Em pacientes com estimulação cardíaca convencional, estáveis clinicamente, a dessincronia intraventricular foi um preditor independente do aumento dos níveis de BNP, após ajuste pela idade e FEVE.

Hatching and larval export of the intertidal crab Neohelice granulata in Mar Chiquita coastal lagoon, Argentina

Synchronization in the events of the reproductive cycle in female Neohelice granulata Dana, 1851 were studied from samples taken weekly and biweekly from September to December 2006 in the Laguna Mar Chiquita. The timing and larval hatching and synchronicity were inferred from numbers of ovigerous females and observing the stages of embryonic development. Synchronization in larval hatching also was observed in females in experiments in dark for a period of 48 hours, at three different salinities (10, 23 and 33 ppm). In addition plankton sampling were performed in order to study larval exportation at the field and its link to the tidal and light/dark cycles. We found that ovigerous females of N. granulata have a marked synchronization in embryonic development which results in that most of berried females are close to hatching within a period of maximum tidal range (days). Within this period, there is a synchronization of hatching at a time scale of hours, governed by environmental conditions. The salinity range used in this study (10-32‰) did not affect hatching synchronicity neither time to hatch. Hatching was synchronized according to endogenous rhythms governed mainly by the tidal cycle and secondarily by the breadth of it. It is also conditioned by the light-dark cycle through an exogenous cycle, so that the hatchings would occur mostly at night high tides.

The use of triphenyltetrazolium chloride in the study of dehydrogenase activity of Brucellae

Experiments for the investigation of dehydrogenase activity of washed cells of a strains of Br. abortus and another of Br. suis in presence of different single added substrates are reported. The activity was measured as the amount of formazan produced by the reduction of 2, 3, 5-triphenyltetrazolum chloride acting as a hydrogen ions acceptor, at pH 7.0. In a general manner the dehydrogenase activity of Br. suis was much more intense than that of Br. abortus (fig. 5). In the conditions of the experiments Br. abortus oxidized L-arabinose, D-galactose, D-glucose, glycerol, D-xylose, DL-alanine, D-fructose, and D-sorbitol. Brucella suis oxidized D-xylose, L-arabinose, D-glucose, D-galactose, DL-alanine, sodium acetate, maltose, glycine, D-fructose, and D-sorbitol. Glycerol was oxidized by Br. abortus but its oxidation by Br. suir was very slight. Sodium acetate and maltose were intensely oxidized by Br. suir but not by Br. abortus. The sites of more intense enzymatic acitivity were seen as small red colored round granules located in one pole of the cells.

Caracterização de sorotipos de Escherichia coli potencialmente produtores de enterotoxinas em alimentos

Em 137 amostras de alimentos de diferentes origens (animal e vegetal) foi investigada a ocorrência de sorotipos de Escherichia coli mais comumente descritos como produtores de enterotoxinas. A análise sorológica dos antígenos somáticos, de envoltórios e flagelares nas 265 culturas isoladas, resultou na identificação de 34 amostras distribuídas em doze sorotipos e oriundas de 24 produtos de origem animal. Outros aspectos foram analisados, visando associá-los como possíveis marcadores epidemiológicos. Assim, na biotipificação, verificou-se o perfil das 34 amostras diante da melibiose, rafinose, sacarose, salicina e sorbitol, obtendo-se a caracterização de 11 biotipos. Todavia, a acentuada heterogeneidade de biotipos distribuídos pelos sorotipos, não permitiu um relacionamento de tipos soro-fermentativos com as fontes de isolamento. Os demais testes, representados pela atividade hemolítica e a capacidade hemaglutinante, pouco acrescentaram para o problema da diferenciação de fenótipos ou na caracterização de marcadores epidemiológicos.