In vitro and in vivo experimental models for drug screening and development for Chagas disease

Chagas disease, a neglected illness, affects nearly 12-14 million people in endemic areas of Latin America. Although the occurrence of acute cases sharply has declined due to Southern Cone Initiative efforts to control vector transmission, there still remain serious challenges, including the maintenance of sustainable public policies for Chagas disease control and the urgent need for better drugs to treat chagasic patients. Since the introduction of benznidazole and nifurtimox approximately 40 years ago, many natural and synthetic compounds have been assayed against Trypanosoma cruzi, yet only a few compounds have advanced to clinical trials. This reflects, at least in part, the lack of consensus regarding appropriate in vitro and in vivo screening protocols as well as the lack of biomarkers for treating parasitaemia. The development of more effective drugs requires (i) the identification and validation of parasite targets, (ii) compounds to be screened against the targets or the whole parasite and (iii) a panel of minimum standardised procedures to advance leading compounds to clinical trials. This third aim was the topic of the workshop entitled Experimental Models in Drug Screening and Development for Chagas Disease, held in Rio de Janeiro, Brazil, on the 25th and 26th of November 2008 by the Fiocruz Program for Research and Technological Development on Chagas Disease and Drugs for Neglected Diseases Initiative. During the meeting, the minimum steps, requirements and decision gates for the determination of the efficacy of novel drugs for T. cruzi control were evaluated by interdisciplinary experts and an in vitro and in vivo flowchart was designed to serve as a general and standardised protocol for screening potential drugs for the treatment of Chagas disease.

Combining ability and heterosis in common bean cultivars

The objective of this work was to determine the combining ability and heterosis, for productivity and yield components, in diallel hybrids derived from crossings between BRSMG-Talismã, IPR Uirapuru, FT Soberano, BRS Campeiro, IAC Tybatã, and IPR Juriti parent cultivars. Fifteen hybrids were generated from diallel crosses, excluding reciprocals. The general and specific combining abilities were significant for plant height, number of pods per plant, number of seeds per plant, number of seeds per pod, 50-seed weight, and grain yield, indicating the occurrence of both additive and nonadditive genetic effects. The best strategy to be adopted is the use of BRS Campeiro, FT Soberano and BRSMG-Talismã cultivars in common bean breeding programs involving selection. The most promising combinations were 'IPR Uirapuru' x 'IAC Tybatã', 'IPR Uirapuru' x 'FT Soberano', 'BRS Campeiro' x 'IPR Juriti', and 'BRS Campeiro' x 'IAC Tybatã'. The parents of these hybrids presented high estimates of specific combining abilities. Hybridization of cultivars belonging to distinguished commercial groups propitiates higher heterosis values in the segregant population.

Morphological, pedigree, and molecular distances and their association with hybrid wheat performance

The objectives of this work were to estimate the genetic distance among wheat genotypes using morphological, pedigree, molecular, and combined morphological and molecular measures, to determine the correlations between these measures, and to evaluate the combining ability of the genotypes. Three generations and two planting designs were studied. Six wheat genotypes were crossed using a diallel design. The F1, F2 and F3generations were evaluated in the field, in the crop seasons of 2003, 2004 and 2005, under spaced plant and full-row planting designs. The estimated general and specific combining abilities of tested hybrids were influenced both by the generation and the planting design. The correlation coefficients among the distance measures and between these measures and genotype performances of different generations for the two planting designs were low to moderate. In order to obtain a more precise estimate of the genetic distance among cultivars and its association with the hybrid performance, more than one generation, planting design, and genetic distance estimation technique should be employed.

Detailed crystallization study of co-precipitated Y1.47Gd1.53Fe5O12 and relevant magnetic properties

The crystallization process of co-precipitated Y1.5Gd1.5Fe5O12 powder heated up to 1000 ºC at rate of 5 °C min-1 was investigated. Above 810 ºC crystalline Y1.47Gd1.53Fe5O12 was obtained with a lattice parameter of 12.41 Å and a theoretical density of 5.84 g cm-3. Dry pressed rings were sintered at 1270 and 1320 ºC, increasing the grain-size from 3.1 to 6.5 µm, the theoretical density by 87.6 to 95.3% and decreasing Hc from 2.9725 to 1.4005 Oe. Additionally, Hc increased when the frequency of the hysteresis graph varied from 60 Hz to 10 kHz, the curie temperature was 282.4 ºC and Ms equalled 9.25 emu g-1 (0.17 kG) agreeing well with the Bs-value of the hysteresis graph and literature values.

HS-SPME-GC-MS ANALYSIS OF VOLATILE AND SEMI-VOLATILE COMPOUNDS FROM DRIED LEAVES OF Mikania glomerata Sprengel

This paper reports on the identification of volatile and semi-volatile compounds and a comparison of the chromatographic profiles obtained by Headspace Solid-Phase Microextraction/Gas Chromatography with Mass Spectrometry detection (HS-SPME-GC-MS) of dried leaves of Mikania glomerata Sprengel (Asteraceae), also known as 'guaco.' Three different types of commercial SPME fibers were tested: polydimethylsiloxane (PDMS), polydimethylsiloxane/divinylbenzene (PDMS/DVB) and polyacrylate (PA). Fifty-nine compounds were fully identified by HS-SPME-HRGC-MS, including coumarin, a marker for the quality control of guaco-based phytomedicines; most of the other identified compounds were mono- and sesquiterpenes. PA fibers performed better in the analysis of coumarin, while PDMS-DVB proved to be the best choice for a general and non-selective analysis of volatile and semi-volatile guaco-based compounds. The SPME method is faster and requires a smaller sample than conventional hydrodistillation of essential oils, providing a general overview of the volatile and semi-volatile compounds of M. glomerata.

Dopaminergic modulation of grooming behavior in virgin and pregnant rats

Dopamine receptors are involved in the expression of grooming behavior. The pregnancy-induced increase in self-licking observed in rats is important for mammary gland development and lactation. This study focuses on the role of dopamine receptor subtypes in grooming behavior of virgin and pregnant female rats. General and mammary gland grooming were measured in virgin rats treated with 0.25 mg/kg of the D1-like agonist SKF-81297 and antagonist SKF-83566 and the D2-like agonist lisuride and antagonist sulpiride. The effects of 0.01 and 0.25 mg/kg doses of the same agonists and antagonists were evaluated in pregnant rats as well. In virgin animals both SKF-83566 and sulpiride treatments significantly reduced the time spent in general grooming, while none of the dopamine agonists was able to significantly change any parameter of general grooming. Time spent in grooming directed at the mammary glands was not affected significantly by any of the drug treatments in virgin rats. All drugs tested significantly decreased the frequency of and the time spent with general grooming, while SKF-81297 treatment alone did not significantly reduce the duration of mammary gland grooming in pregnant rats. These data show that in female rats the behavioral effects of D1-like and D2-like dopamine receptor stimulation and blockade differ according to physiological state. The results suggest that dopamine receptors may play specific roles modulating grooming behavior in pregnant rats. Since grooming of the mammary gland during pregnancy may influence lactation, this aspect is relevant for studies regarding the perinatal use of dopamine-related drugs.

Sialic acid changes in Dalton's lymphoma-bearing mice after cyclophosphamide and cisplatin treatment

Sialic acid changes in Dalton's lymphoma cells and other tissues of 10-12-week-old Swiss albino mice were investigated in relation to tumour growth in vivo and following cyclophosphamide (ip, 200 mg/kg body weight) or cisplatin (ip, 8 mg/kg body weight) treatment. Three to four animals of both sexes were used in each experimental group. The sialic acid level of tumour cells (0.88 µmol/g) increased with tumour progression (1.44-1.59 µmol/g; P<=0.05) in mice. Sialic acid concentration in other tissues (liver, kidney, testes and brain) also increased (~40, 10, 30 and 58%, respectively) in the tumour-bearing hosts as compared with that in the respective tissues of normal mice. In vivo cyclophosphamide or cisplatin treatment resulted in an overall decrease of sialic acid contents in the tissues. Cyclophosphamide was more efficient in lowering tissue sialic acid than cisplatin (P<=0.01, ANOVA). It is suggested that sialic acid residues could be an important factor contributing to the manifestation of malignant properties in cancer cells in general and Dalton's lymphoma cells in particular. A significant decrease in the sialic acid content of Dalton's lymphoma cells after cisplatin or cyclophosphamide treatment may bring about specific changes in tumour cells which could be associated with tumour regression.

Somaclonal variation: a morphogenetic and biochemical analysis of Mandevilla velutina cultured cells

Cell cultures of Mandevilla velutina have proved to be an interesting production system for biomass and secondary metabolites able to inhibit the hypotensive activity of bradykinin, a nonapeptide generated in plasma during tissue trauma. The crude ethyl acetate extract of cultured cells contains about 31- to 79-fold more potent anti-bradykinin compounds (e.g., velutinol A) than that obtained with equivalent extracts of tubers. Somaclonal variation may be an explanation for the wide range of inhibitor activity found in the cell cultures. The heterogeneity concerning morphology, differentiation, carbon dissimilation, and velutinol A production in M. velutina cell cultures is reported. Cell cultures showed an asynchronous growth and cells in distinct developmental stages. Meristematic cells were found as the major type, with several morphological variations. Cell aggregates consisting only of meristematic cells, differentiated cells containing specialized cell structures such as functional chloroplasts (cytodifferentiation) and cells with embryogenetic characteristics were observed. The time course for sucrose metabolism indicated cell populations with significant differences in growth and metabolic rates, with the highest biomass-producing cell line showing a cell cycle 60% shorter and a metabolic rate 33.6% higher than the control (F2 cell population). MALDI-TOF mass spectrometric analysis of velutinol A in selected cell lines demonstrated the existence of velutinol A producing and nonproducing somaclones. These results point to a high genetic heterogeneity in general and also in terms of secondary metabolite content.