Salt-induced changes on H<Superscript>+</Superscript>-ATPase activity,sterol and phospholipid content and lipid peroxidation of root plasma membrane from dwarf-cashew (<Emphasis Type="Italic">Anacardium occidentale</Emphasis> L.) seedlings

The effects of NaCl stress on the growth, water relation, gas exchange, tissue mineral content, and on H⁺-ATPase activity, lipid composition and peroxidation of root plasma membrane-enriched fractions of two genotypes (CCP06 and BRS189, sensitive and tolerant to salt stress, respectively) of dwarf-precocious cashew were studied. Growth reduction was higher in CCP06 than in BRS189. Net photosynthesis decreased in both genotypes, CCP06 being more affected. Roots of BRS189 accumulated higher amount of Na⁺ than those of CCP06 at both salt treatments, whereas Cl⁻ increase was higher only at 8 dS m⁻¹. NaCl at 8 dS m⁻¹ did not modify the plasma membrane H⁺-ATPase activity in CCP06 roots, but significantly increased it in BRS189 roots. Lipid peroxidation was lower in BRS189 than in CCP06 roots. Salinity induced higher accumulation of proline in BRS189 roots. Total phospholipids and free sterols content increased significantly in root plasma membrane of CCP06. However, in BRS189, a slight reduction of free sterols content and no changes in total phospholipids content were observed. Thus, the results suggest that the ability of cashew seedlings to adapt to salt stress is, at least in part, dependent upon the maintenance of integrity and protection against oxidative damage of plasma membrane, which could favor the activation of plasma membrane H⁺-ATPase, as a cellular mechanism to regulate ion exclusion from the shoot.     

Leguminous lectins as tools for studying the role of sugar residues in leukocyte recruitment

The natural physiological ligands for selectins are oligosaccharides found in glycoprotein or glycolipid molecules in cell membranes. In order to study the role of sugar residues in the in vivo lectin anti-inflammatory effect, we tested three leguminous lectins with different carbohydrate binding affinities in the peritonitis and paw oedema models induced by carrageenin in rats. L. sericeus lectin was more anti-inflammatory than D. virgata lectin, the effects being reversed by their specific binding sugars (N-acetylglucosamine and alpha-methylmannoside, respectively). However, V. macrocarpa, a galactose-specific lectin, was not anti-inflammatory. The proposed anti-inflammatory activity of lectins could be due to a blockage of neutrophil-selectin carbohydrate ligands. Thus, according to the present data, we suggest an important role for N-acetylglucosamine residue as the major ligand for selectins on rat neutrophil membranes.     

Antimalarial drug susceptibility testing of Plasmodium falciparum in Brazil using a radioisotope method

From March 1996 to August 1997, a study was carried out in a malaria endemic area of the Brazilian Amazon region. In vivo sensitivity evaluation to antimalarial drugs was performed in 129 patients. Blood samples (0.5 ml) were drawn from each patient and cryopreserved to proceed to in vitro studies. In vitro sensitivity evaluation performed using a radioisotope method was carried out with the cryopreserved samples from September to December 1997. Thirty-one samples were tested for chloroquine, mefloquine, halofantrine, quinine, arteether and atovaquone. Resistance was evidenced in 96.6% (29/30) of the samples tested for chloroquine, 3. 3% (1/30) for quinine, none (0/30) for mefloquine and none for halofantrine (0/30). Overall low sensitivity was evidenced in 10% of the samples tested for quinine, 22.5% tested for halofantrine and in 20% tested for mefloquine. Means of IC 50 values were 132.2 (SD: 46. 5) ng/ml for chloroquine, 130.6 (SD: 49.6) ng/ml for quinine, 3.4 (SD: 1.3) ng/ml for mefloquine, 0.7 (SD: 0.3) ng/ml for halofantrine, 1 (SD: 0.6) ng/ml for arteether and 0.4 (SD: 0.2) ng/ml for atovaquone. Means of chloroquine IC 50 of the tested samples were comparable to that of the chloroquine-resistant strain W2 (137.57 ng/ml) and nearly nine times higher than that of the chloroquine-sensitive strain D6 (15.09 ng/ml). Means of quinine IC 50 of the tested samples were 1.7 times higher than that of the low sensitivity strain W2 (74.84 ng/ml) and nearly five times higher than that of the quinine-sensitive strain D6 (27.53 ng/ml). These results disclose in vitro high resistance levels to chloroquine, low sensitivity to quinine and evidence of decreasing sensitivity to mefloquine and halofantrine in the area under evaluation.     

Differences in aerial and terrestrial visual scanning in captive black tufted-ear marmosets (Callithrix penicillata) exposed to a novel environment

Aerial and terrestrial visual scanning were investigated in captive black tufted-ear marmosets, Callithrix penicillata, exposed to a novel environment. Naive adult subjects (n=24) were individually exposed to a figure-eight maze during seven 30-min trials, 48 h apart. Habituation to the maze was observed, as indicated by the significant decrease in locomotion. The frequency of aerial scanning, however, remained elevated throughout the 7 trials, while its duration rapidly increased to high levels. Frequency and duration of terrestrial scanning persisted at constant low rates, differing significantly from aerial scanning. Males and females did not differ significantly. The different impact of aerial versus terrestrial predators could have a significant influence on vigilant behaviour in this species. Thus, visual scanning is an important and highly organized antipredation strategy in marmosets.     

Drug susceptibility of Brazilian strains of Mycobacterium bovis using traditional and molecular techniques

Transmission of Mycobacterium bovis from cattle to humans has been reported and can cause tuberculosis (Tb) and a problem in certain risk populations. Therefore, knowledge of resistance of M. bovis towards antibiotics used for therapy of human Tb could help avoiding cure delay and treatment cost increase when dealing with drug resistant organisms. We therefore evaluated the susceptibility of M. bovis isolates towards streptomycin, isoniazide, rifampicin, ethambutol, and ethionamide, the first line antibiotics for human Tb. Therefore, 185 clinical samples from cattle with clinical signs of tuberculosis were processed and submitted to culturing and bacterial isolates to identification and drug susceptibility testing using the proportion method. Among 89 mycobacterial strains, 65 were identified as M. bovis and none were resistant to any of the antibiotics used. Confirmation of present results by future studies, enrolling a large number of isolates and designed to properly represent Brazilian regions, may favor the idea of using isoniazide preventive therapy as part of a Tb control strategy in special situations. Also, nucleic acids from bacterial isolates were submitted to rifoligotyping, a recently described reverse hybridization assay for detection of mutations causing resistance towards rifampicin. Concordance between the conventional and the molecular test was 100%, demonstrating the use of such methodology for rapid evaluation of drug susceptibility in M. bovis.     

Miniaturized multichannel near infrared endoscope for mouse imaging

We describe the design and construction of a miniaturized multichannel near infrared (NIR) endoscopic imaging system developed for high-resolution imaging of mice. The device allows for simultaneous real-time video images in white light and two independent NIR channels. Testing demonstrated independent acquisition of nanomolar concentrations of fluorochromes Cy5.5 and Cy7. Cross-talk between the NIR channels, partially a result of broad tails in the spectra of commonly used organic fluorochromes, was assessed, modeled for the linear range of the concentration/signal intensity function, and compensated. The calculated compensation was 5.5% and 22% of the total signal intensity in the two channels NIR700 and NIR780, respectively, at equal concentrations of the two fluorochromes. Using a mouse model of colonic adenomatosis, we show that both perfusion and protease activity can be detected simultaneously, independently, and repeatedly in live mice. The developed device should be useful for in vivo imaging of diverse molecular targets.     

Evidence that intrinsic iron but not intrinsic copper determines S-nitrosocysteine decomposition in buffer solution.

The present experiments were designed to analyze the influence of copper and iron ions on the process of decomposition of S-nitrosocysteine (cysNO), the most labile species among S-nitrosothiols (RSNO). CysNO fate in buffer solution was evaluated by optical and electron paramagnetic resonance (EPR) spectroscopy, and the consequences on its vasorelaxant effect were studied on noradrenaline-precontracted rat aortic rings. The main results are the following: (i) copper or iron ions, especially in the presence of the reducing agent ascorbate, accelerated the decomposition of cysNO and markedly attenuated the amplitude and duration of the relaxant effect of cysNO; (ii) by contrast, the iron and copper chelators bathophenantroline disulfonic acid (BPDS) and bathocuproine disulfonic acid (BCS) exerted a stabilizing effect on cysNO, prolonged its vasorelaxant effect, and abolished the influence of ascorbate; (iii) in the presence of ascorbate, BPDS displayed a selective inhibitory effect toward the influence of iron ions (but not toward copper ions) on cysNO decomposition and vasorelaxant effect, while BCS prevented the effects of both copper and iron ions; (iv) L-cysteine enhanced stability and prolonged the relaxant effect of cysNO; (v) the process of iron-induced decomposition of cysNO was associated with the formation of EPR-detectable dinitrosyl-iron complexes (DNIC) either with non-thiol- or thiol-containing ligands (depending on the presence of L-cysteine), both of which exhibiting vasorelaxant properties. From these data, it is concluded that the amount of intrinsic copper was probably too low to produce a destabilizing effect even on the most labile RSNO, cysNO, and that only intrinsic iron, through the formation of DNIC, was responsible for the process of cysNO decomposition and thus influenced its vasorelaxant properties.     

Morphometric comparison of Simulium perflavum larvae (Diptera: Simuliidae) in relation to season and gender in Central Amazônia, Brazil

Number of larval instars, age structure and environmental effects on these parameters represent basic information in the study of insect population biology. When species have economic importance, this information is essential in order to choose the best period to apply different control methods and to determine the stages of the life cycle of the insect that are most susceptible to each treatment. The family Simuliidae has many species of medical/veterinary importance in the world, and some studies in the temperate region have suggested that the number of larval instars and the larval size can vary according to the season, gender and some environmental factors, such as temperature and diet. This study, with the zoophilic species Simulium perflavum Roubaud, is the first in the Neotropics observing some of these factors and will serve as a template for other species of medical importance in the region. S. perflavum larvae were collected in five streams in Central Amaz?nia (Manaus and Presidente Figueiredo counties, State of Amazonas), in Sept./Oct. 1996 (dry season) and Feb./Mar. 1997 (rainy season). These larvae were measured (lateral length of head capsule and width of cephalic apodema) to determine the number of larval instars (n=3985), to compare the larval size between seasons and genders (last and penultimate larval instars, n=200). Seven larval instars were determined for this species using frequency distributions, t-tests and Crosbys growth rule. Significant differences were not detected (t-test, p>0.05) in larval size between seasons and genders. Our results differ from some found in temperate regions suggesting that in the Neotropical region the larval size in different seasons and different genders remains constant, although some environmental parameters, such as diet, change depending on the season.     

Stomach content analyses of Simulium perflavum Roubaud 1906 (Diptera: Simuliidae) larvae from streams in Central Amazônia, Brazil

Stomach contents of Simulium perflavum Roubaud larvae were analyzed and compared with plankton and periphyton collected in five streams, in Central Amazonia (Manaus and Presidente Figueiredo counties), in Sep./Oct.1996 (dry season) and Feb./Mar. 1997 (rainy season). A total of 1,400 last-instar larvae were dissected; the stomach contents were analyzed using different methods: fresh and after oxidation. A total of 87 taxa (algae, diatoms and rotifers) were found in the stomach contents. In each stream, qualitative samples of plankton and periphyton were collected; these were mounted between slides and cover slips. A total of 94 taxa of plankton and 54 taxa of periphyton were collected. One species of Rotifera was present in the stomach contents, plankton and periphyton. Cluster analysis based on species composition of the organisms present in the stomach contents grouped the streams into two major groups, each belonging to a different drainage area. Correlations based on presence/absence of species of microalgae in the stomach contents, plankton and periphyton indicated significant associations (p<0.05) between stomach contents and plankton and between plankton and periphyton (z test); the Sorensen coefficient and cluster analysis corroborate the same associations.