Rheotaxis of Biomphalaria glabrata on vertical substrates and its role in the recolonization of habitats treated with molluscicides

The authors observed specimens of Biomphalaria glabrata climbing up the vertical wall of a ditch against the current. The snails that showed this behavior during application of a molluscicide in the breeding site survived and probably played a role in repopulation, which was observed three months later. These observations motivated field and laboratory investigations which led the authors to conclude that: a) this species is able to climb vertical surfaces both in field and laboratory situations; b) the current of water, as a physical stimulus, is sufficient to trigger this behavior (rheotaxis); c) rheotaxis on vertical surfaces depends on the presence of a necessarily moderate current; d) there are indications that B. glabrata may undergo habituation with respect to rheotaxis on vertical walls, e) the relationship between rheotaxis and habituation should be considered as a factor causing snail grouping in water bodies which may contribute to their localization in the field; f) rheotaxis on vertical surfaces may facilitate population dispersal, and its occurrence should be considered when campaigns for the control of schistosomiasis transmission are planned. The authors present some proposals to avoid the manifestation of this behavior in some filed situations.

Simultaneous LC-MS/MS quantification of P-glycoprotein and cytochrome P450 probe substrates and their metabolites in DBS and plasma.

BACKGROUND: An LC-MS/MS method has been developed for the simultaneous quantification of P-glycoprotein (P-gp) and cytochrome P450 (CYP) probe substrates and their Phase I metabolites in DBS and plasma. P-gp (fexofenadine) and CYP-specific substrates (caffeine for CYP1A2, bupropion for CYP2B6, flurbiprofen for CYP2C9, omeprazole for CYP2C19, dextromethorphan for CYP2D6 and midazolam for CYP3A4) and their metabolites were extracted from DBS (10 µl) using methanol. Analytes were separated on a reversed-phase LC column followed by SRM detection within a 6 min run time. RESULTS: The method was fully validated over the expected clinical concentration range for all substances tested, in both DBS and plasma. The method has been successfully applied to a PK study where healthy male volunteers received a low dose cocktail of the here described P-gp and CYP probes. Good correlation was observed between capillary DBS and venous plasma drug concentrations. CONCLUSION: Due to its low-invasiveness, simple sample collection and minimal sample preparation, DBS represents a suitable method to simultaneously monitor in vivo activities of P-gp and CYP.

Pathological substrates of cognitive decline in Alzheimer's disease

The progressive development of Alzheimer's disease (AD)-related lesions such as neurofibrillary tangles,amyloid deposits and synaptic loss within the cerebral cortex is a main event of brain aging.Recent neuropathologic studies strongly suggested that the clinical diagnosis of dementia depends more on the severity and topography of pathologic changes than on the presence of a qualitative marker. However, several methodological problems such as selection biases, case-control design,density-based measures, and masking effects of concomitant pathologies should be taken into account when interpreting these data. In last years, the use of stereologic counting permitted to define reliably the cognitive impact of AD lesions in the human brain. Unlike fibrillar amyloid deposits that are poorly or not related to the dementia severity, the use of this method documented that total neurofibrillary tangles and neuron numbers in the CA1 field are the best correlates of cognitive deterioration in brain aging. Loss of dendritic spines in neocortical but not hippocampal areas has a modest but independent contribution to dementia. In contrast, the importance of early dendritic and axonal tau-related pathologic changes such as neuropil threads remains doubtful. Despite these progresses, neuronal pathology and synaptic loss in cases with pure AD pathology cannot explain more than 50% of clinical severity. The present review discusses the complex structure/function relationships in brain aging and AD within the theoretical framework of the functional neuropathology of brain aging.

Peptide substrate specificity of the membrane-bound metalloprotease of Leishmania.

The promastigote surface protease (PSP) of Leishmania is a neutral membrane-bound zinc enzyme. The protease has no exopeptidase activity and does not cleave a large selection of substrates with chromogenic and fluorogenic leaving groups at the P1' site. The substrate specificity of the enzyme was studied by using natural and synthetic peptides of known amino acid sequence. The identification of 11 cleavage sites indicates that the enzyme preferentially cleaves peptides at the amino side when hydrophobic residues are in the P1' site and basic amino acid residues in the P2' and P3' sites. In addition, tyrosine residues are commonly found at the P1 site. Hydrolysis is not, however, restricted to these residues. These results have allowed the synthesis of a model peptide, H2N-L-I-A-Y-L-K-K-A-T-COOH, which is cleaved by PSP between the tyrosine and leucine residues with a kcat/Km ratio of 1.8 X 10(6) M-1 s-1. Furthermore, a synthetic nonapeptide overlapping the last four amino acids of the prosequence and the first five residues of mature PSP was found to be cleaved by the protease at the expected site to release the mature enzyme. This result suggests a possible autocatalytic mechanism for the activation of the protease. Finally, the hydroxamate-derivatized dipeptide Cbz-Tyr-Leu-NHOH was shown to inhibit PSP competitively with a KI of 17 microM.

Capture of Caliptrate flies with different breeding substrates on beaches in Rio de Janeiro, RJ, Brazil

Muscidae flies belonging to four Familia and 13 species in a total number of 3.652 specimens were collected from beaches at Ilha do Governador, Rio de Janeiro, Brazil using different breeding substrates, and subsequently bred in the laboratory. Captures were done from April to November 1989, using in a first phase different substrates: fruits (banana and papaya), vegtable (tomato), animal viscera (bovine liver), marine animals (fish, crab, shrimp, squid), mouse carcass and feaces (human and canine). The species collected more often were: Fannia sp. (subgroup pusio), Chrysomya megacephala, Phaenicia eximia, Synthesiomyia nudiseta, Peckya chrysostoma, Musca domestica and Atherigona orientalis. In a later phase, only fish was used, as bait and placed directly on the beach sand. From a total of 189 pupae, the following adult specimen were obtained: Peckia chrysostoma (58.06%), Chrysomya megacephala (30.64%) and in lesser numbers Synthesiomyia nudiseta and Phaenicia eximia.

Ovipositional substrates used by Calyptrate Diptera in Tijuca forest, Rio de Janeiro

Twenty seven species of calyptrate muscoids were reared from a forested area of Rio de Janeiro (Tijuca Forest). Substrates for obtaining flies were beef liver, fish, mouse, frog, shrimp, snail carcasses, human faeces, banana and papaya fruits. The most frequent species found were: Fannia sp. (subgroup pusio) (49.9% on shrimp). Hemilucilia flavifacies (95.0% on liver). Phaenicia eximia (49.4% on mouse), Synthesiomyia nudiseta ( 100.0% on fish), Ophyra aenescens (100.0% on shrimp), Oxyvinia excisa (100.0% on faeces), Euboettecheria collusor (52.4% on faeces) and Pattonella intermutans (61.0% on frog).

Neuropathological substrates and structural changes in late-life depression: the impact of vascular burden.

A first episode of depression after 65 years of age has long been associated with both severe macrovascular and small microvascular pathology. Among the three more frequent forms of depression in old age, post-stroke depression has been associated with an abrupt damage of cortical circuits involved in monoamine production and mood regulation. Late-onset depression (LOD) in the absence of stroke has been related to lacunes and white matter lesions that invade both the neocortex and subcortical nuclei. Recurrent late-life depression is thought to induce neuronal loss in the hippocampal formation and white matter lesions that affect limbic pathways. Despite an impressive number of magnetic resonance imaging (MRI) studies in this field, the presence of a causal relationship between structural changes in the human brain and LOD is still controversial. The present article provides a critical overview of the contribution of neuropathology in post-stroke, late-onset, and late-life recurrent depression. Recent autopsy findings challenge the role of stroke location in the occurrence of post-stroke depression by pointing to the deleterious effect of subcortical lacunes. Despite the lines of evidences supporting the association between MRI-assessed white matter changes and mood dysregulation, lacunes, periventricular and deep white matter demyelination are all unrelated to the occurrence of LOD. In the same line, neuropathological data show that early-onset depression is not associated with an acceleration of aging-related neurodegenerative changes in the human brain. However, they also provide data in favor of the neurotoxic theory of depression by showing that neuronal loss occurs in the hippocampus of chronically depressed patients. These three paradigms are discussed in the light of the complex relationships between psychosocial determinants and biological vulnerability in affective disorders.

Comparative evaluation of pyrethroid insecticide formulations against Triatoma infestans (Klug): residual efficacy on four substrates

We investigated the residual efficacy of four insecticide formulations used in Chagas disease vector control campaigns: cyfluthrin 12.5% suspension concentrace (SC), lambda-cyhalothrin 10% wettable powder (WP), deltamethrin 2.5% SC, and 2.5% WP on four types of circular blocks of wood, straw with mud, straw with mud painted with lime, and mud containing 5% of cement. Three concentrations of these insecticides were tested: the LC90 (previously determined on filter paper), the double of the LC90, and the recommended operational dose. For each bioassay test, 15 third-stage nymphs of Triatoma infestans (Klug) (Hemiptera: Reduviidae) were exposed for 120 h to each treatment at 24 h, 30, 60, 90, and 180 days post-spraying. Mortality rates, moulting history and behaviour were recorded at 24, 48, 72, and 120 h of exposure. Mortality rates were highest during the first 30 days post-spraying. Highest mortality rates (above 50%) were observed for deltamethrin 2.5% SC and lambda-cyhalothrin 10% WP on wood blocks up to three months post-spraying. Mud was the substrate on which treatments showed lowest persistence, with the other two substrates showing intermediate residual efficacy of all treatments. During the first 30 days WP formulations were not as effective as SC flowable formulations but, overall in the longer term, WP gave grater mortality rates of T. infestans nymphs exposed at up to six months post-spraying. Porous surfaces, especially mud, showed most variability presumably due to absorption of the insecticide. In contrast the less porous surfaces (i.e. wood and lime-coated mud) kept mortality rates high for longer post-treatment, irrespective of the insecticide concentration used.

Caspase substrates and neurodegenerative diseases.

Apoptosis is induced by the cleavage of a subset of cellular proteins by proteases of the caspase family. Numerous (hundreds) caspase substrates have been described but only for a few of them is the function of their cleavage by caspases well understood. In this review, apoptosis and caspases will first be introduced. The main focus will then be directed to the caspase substrates, the actual "workers" doing the job of mediating and regulating the apoptotic process. The caspase substrates whose functions upon cleavage have been carefully investigated and those that are potentially involved in neurodegenerative diseases will be discussed in detail.

Micro-patterning of biomolecules on polymer substrates.

UV−excimer laser photoablation was used, in combination with surface blocking techniques, to pattern proteins on the surfaces of polyimide and poly(ethylene terephthalate). This technique involves physical adsorption of avidin through laser-defined openings in low-temperature laminates or adsorbed protein blocking layers. Visualization of biomolecular patterns were monitored using avidin and fluorescein-labeled biotin as a model receptor−ligand couple. Adsorbed proteins could be shown to bind to UV-laser-treated polymer surfaces up to three times higher than on commercially available polymers. UV-laser photoablation was also used for the generation of three-dimensional structure, which leads to the possibility of biomolecule patterning within polymer-based microanalytical systems. The simplicity and easy handling of the described technique facilitate its application in microdiagnostic devices.

The adequacy of artificial oviposition substrates for laboratory rearing of Piezodorus guildinii (Westwood) (Heteroptera, Pentatomidae)

Studies in the laboratory tested the suitability of synthetic wool string, cotton string, cheesecloth, and commercial cotton ball as artificial oviposition substrates for the small green stink bug, Piezodorus guildinii (Westwood) (Heteroptera, Pentatomidae). In confined cages, 54% of total egg masses was laid on synthetic wool string, 31% on cotton string, and only 15% on cheesecloth. In an additional test, the best substrate selected, synthetic wool string, received 92% of egg masses compared to 8% on the commonly used substrate, cotton ball. Synthetic wool string received the most egg masses of any size, in particular those in the range 11-20 eggs/mass. Because the eggs of P. guildinii are laid in two parallel double rows, the egg masses fit the wool string perfectly.

Use of artificial substrates of different colors for oviposition by the brown stink bug Euschistus heros (Hemiptera, Pentatomidae)

Aiming to contribute to a rearing methodology for the brown stink bug, Euschistus heros, in the laboratory, we evaluated oviposition on artificial substrates of different colors. During six days, oviposition was evaluated daily, by counting the total number of eggs, number of clutches, and eggs/clutch. Females laid 12,463 eggs, in 1,677 clutches, resulting in an average of 7.28 ± 0.44 eggs/clutch. Black, brown, and green felt had the most eggs and clutches. The results demonstrated that many colors are suitable as oviposition substrate for E. heros, providing information for the mass rearing of this insect.

Knock, knock, knocking on muscle doors. Visions on the transport of substrates across the plasma membrane in muscle.

Muscle is a major player in metabolism. It uses large amounts of glucose in the absorptive state and changes in muscle insulin-stimulated glucose uptake alter whole-body glucose disposal. Lipid substrates such as fatty acids or ketone bodies are preferentially used by muscle in certain physiological conditions. Muscle is also the main reservoir of amino acids and protein. The activity of many different plasma membrane transporters such as glucose carriers, carnitine, creatine or amino acid transporters maintain muscle metabolism by taking up or releasing substrates or metabolites across the cell surface. The goal of this review is the molecular characterization of muscle membrane transporter proteins and the analysis of their regulatory roles.

Screen printed La2/3Sr1/3MnO3 thick films on alumina substrates

We report here on the preparation of La2/3Sr1/3MnO3 magnetoresistive thick films on polycrystalline Al2O3 substrates by using the screen printing technique. It is shown that films can be obtained using high temperature sintering. While there is a reacted layer, this improves adhesion and is not too troublesome if the films are made thick enough. It is shown that PbO-B2O3-SiO2 glass additives allow sintering at lower temperatures and can be used to improve the mechanical stress of the films. However, it is found that glass concentrations large enough to significantly improve the film adherence result in a weak low field magnetoresistance probably because grains are coated with high resistivity material. Strategies to overcome these difficulties are discussed.