A Multiplex-PCR approach to identification of the Brazilian intermediate hosts of Schistosoma mansoni

Due to difficulties concerning morphological identification of planorbid snails of the genus Biomphalaria, and given a high variation of characters and in the organs with muscular tissue, we designed specific polymerase chain reaction (PCR) primers for Brazilian snail hosts of Schistosoma mansoni from available sequences of internal transcribed spacer 2 (ITS2) of the ribosomal RNA gene. From the previous sequencing of the ITS2 region, one primer was designed to anchor in the 5.8S conserved region and three other species-specific primers in the 28S region, flanking the ITS2 region. These four primers were simultaneously used in the same reaction (Multiplex-PCR), under high stringency conditions. Amplification of the ITS2 region of Biomphalaria snails produced distinct profiles (between 280 and 350 bp) for B. glabrata, B. tenagophila and B. straminea. The present study demonstrates that Multiplex-PCR of ITS2-DNAr showed to be a promising auxiliary tool for the morphological identification of Biomphalaria snails, the intermediate hosts of S. mansoni.

Classification of immature mosquito species according to characteristics of the larval habitat in the subtropical province of Chaco, Argentina

To classify mosquito species based on common features of their habitats, samples were obtained fortnightly between June 2001-October 2003 in the subtropical province of Chaco, Argentina. Data on the type of larval habitat, nature of the habitat (artificial or natural), size, depth, location related to sunlight, distance to the neighbouring houses, type of substrate, organic material, vegetation and algae type and their presence were collected. Data on the permanence, temperature, pH, turbidity, colour, odour and movement of the larval habitat's water were also collected. From the cluster analysis, three groups of species associated by their degree of habitat similarity were obtained and are listed below. Group 1 consisted of Aedes aegypti. Group 2 consisted of Culex imitator, Culex davisi, Wyeomyia muehlensi and Toxorhynchites haemorrhoidalis separatus. Within group 3, two subgroups are distinguished: A (Psorophora ferox, Psorophora cyanescens, Psorophora varinervis, Psorophora confinnis, Psorophora cingulata, Ochlerotatus hastatus-oligopistus, Ochlerotatus serratus, Ochlerotatus scapularis, Culex intrincatus, Culex quinquefasciatus, Culex pilosus, Ochlerotatus albifasciatus, Culex bidens) and B (Culex maxi, Culex eduardoi, Culex chidesteri, Uranotaenia lowii, Uranotaenia pulcherrima, Anopheles neomaculipalpus, Anopheles triannulatus, Anopheles albitarsis, Uranotaenia apicalis, Mansonia humeralis and Aedeomyia squamipennis). Principal component analysis indicates that the size of the larval habitats and the presence of aquatic vegetation are the main characteristics that explain the variation among different species. In contrast, water permanence is second in importance. Water temperature, pH and the type of larval habitat are less important in explaining the clustering of species.

Descriptions of new species with a key to identification of the genus Neodexiopsis Malloch (Diptera, Muscidae) in Brazil

Nine new species of Neodexiopsis Malloch from Paraná, southern Brazil, are described: Neodexiopsis cinerea sp. nov. and N. paranaensis sp. nov. from Ponta Grossa; N. facilis sp. nov., N. legitima sp. nov., N. similis sp. nov. and N. uber sp. nov. from Guarapuava; N. erecta sp. nov., N. pura sp. nov. and N. rara sp. nov. from Colombo. A key to the identification of the Brazilian species of Neodexiopsis is also presented.

La diminution de l'activité du répresseur ICER dans les adipocytes est responsable de la résistance à l'insuline dirigée par le facteur CREB dans l'obésité [The decreased activity of the repressor ICER in adipocytes is responsible for insulin resistance led by factor CREB in obesity.]

Introduction: Une élévation de l'activité des facteurs de transcription CREBs dans le tissu adipeux est en partie responsable de l'insulino-résistance systémique dans l'obésité. Le facteur «Inducible cAMP early repressor» (ICER) est un répresseur transcriptionnel passif dont le niveau d'expression antagonise l'activité des CREBs. L'objectif de ce travail adipocytaire des CREBs dans l'obésité chez l'Homme et la souris. Matériels et méthodes: Du tissu adipeux blanc (TAB) a été prélevé chez des souris obèses nourries sous une diète normale et des souris obèses nourries sous un régime riche en graisses pendant 12 semaines. Des biopsies de tissu adipeux viscéral (TAV) ont été prélevées chez les sujets humains minces (BMI = 24 ± 0,5 kg/m2) et obèses (BMI > 35 kg/m2). L'expression des gènes est quantifiée par RT-PCR quantitative. L'activité des CREBs et d'ICER est mesurée par des expériences de retard sur gel. L'activité des histones déacétylases est quantifiée par dosage colorimétrique. Résultats: L'expression et l'activité d'ICER sont diminuées dans le TAB des souris obèses, hyper-glycémiques et insulino-résistantes. De même, l'activité d'ICER est réduite dans le TAV des sujets humains obèses. Cette réduction corrèle avec une augmentation de l'activité des CREBs, une réduction de l'expression de Glut4 et de l'adiponectine, à la fois chez l'Homme et la souris. La diminution de l'expression d'ICER n'est observée que dans la fraction adipocytaire du tissu adipeux. L'expression d'ICER est contrôlée par l'activité des HDACs. L'inhibition des HDACs inhibe l'expression d'ICER dans les adipocytes. L'activité totale des HDACs est réduite dans les tissus adipeux chez les souris et chez les sujets humains obèses. Conclusion: La diminution de l'activité d'ICER dans les adipocytes par une modification de l'activité des HDACs serait responsable de l'augmentation de l'activité des CREBs dans l'obésité.

Treatment of rats with a self-selected hyperlipidic diet, increases the lipid content of the main adipose tissue sites in a proportion similar to that of the lipids in the rest of organs and tissues

Adipose tissue (AT) is distributed as large differentiated masses, and smaller depots covering vessels, and organs, as well as interspersed within them. The differences between types and size of cells makes AT one of the most disperse and complex organs. Lipid storage is partly shared by other tissues such as muscle and liver. We intended to obtain an approximate estimation of the size of lipid reserves stored outside the main fat depots. Both male and female rats were made overweight by 4-weeks feeding of a cafeteria diet. Total lipid content was analyzed in brain, liver, gastrocnemius muscle, four white AT sites: subcutaneous, perigonadal, retroperitoneal and mesenteric, two brown AT sites (interscapular and perirenal) and in a pool of the rest of organs and tissues (after discarding gut contents). Organ lipid content was estimated and tabulated for each individual rat. Food intake was measured daily. There was a surprisingly high proportion of lipid not accounted for by the main macroscopic AT sites, even when brain, liver and BAT main sites were discounted. Muscle contained about 8% of body lipids, liver 1-1.4%, four white AT sites lipid 28-63% of body lipid, and the rest of the body (including muscle) 38-44%. There was a good correlation between AT lipid and body lipid, but lipid in"other organs" was highly correlated too with body lipid. Brain lipid was not. Irrespective of dietary intake, accumulation of body fat was uniform both for the main lipid storage and handling organs: large masses of AT (but also liver, muscle), as well as in the"rest" of tissues. These storage sites, in specialized (adipose) or not-specialized (liver, muscle) tissues reacted in parallel against a hyperlipidic diet challenge. We postulate that body lipid stores are handled and regulated coordinately, with a more centralized and overall mechanisms than usually assumed.

Chemical composition and antimicrobial activity of the essential oil of Hyptis pectinata (l.) Poit.

Essential oil was extracted from leaves of Hyptis pectinata using hydrodistillation, and its composition determined using GC-FID and GC-MS. Chemical analysis showed that there was a predominance of sesquiterpenes, of which β-caryophyllene (18.34%), caryophyllene oxide (18.00%) and calamusenone (24.68%) were measured for the first time in the genus Hyptis. Twenty-one compounds were identified, and calamusenone was isolated using preparative thin layer chromatography with a silica gel plate (60 PF254). The minimal inhibitory concentration (MIC) and minimal microbicidal concentration (MMC) were determined for various pathogenic microorganisms. H. pectinata oil was most effective against Gram (+) bacteria and yeasts.

Chemical composition and analgesic activity of the leaves and branches of Marlierea tomentosa Camb.

This paper describes the isolation of five phytoconstituents from Marlierea tomentosa. The triterpenes α-amyrin (1) and β-amyrin (2), and the flavonoids quercitrin (3) and isoquercetin (4), were isolated from the leaves. The branches afforded the triterpene arjunolic acid (5). The antinociceptive activity of crude extract, fractions and compound 5 were tested using the writhing and formalin tests in mice. The crude extract, some fractions, particularly dichloromethane and butanol (leaves), ethyl acetate (branches) and arjunolic acid (5) (81.2% inhibition) were more active against the writhing test than the two reference drugs, acetylsalicylic acid and indomethacin.

Mathematical model to estimate of the deterioration of wooden poles in contact with soil used in rural areas

In São Paulo State, mainly in rural areas, the utilization of wooden poles is observed for different purposes. In this context, wood in contact with the ground presents faster deterioration, which is generally associated to environmental factors and, especially to the presence of fungi and insects. With the use of mathematical models, the useful life of wooden structures can be predicted by obtaining "climatic indexes" to indicate, comparatively among the areas studied, which have more or less tendency to fungi and insects attacks. In this work, by using climatological data of several cities at São Paulo State, a simplified mathematical model was obtained to measure the aggressiveness of the wood in contact with the soil.

Central 5-HT2A receptors modulate the vagal bradycardia in response to activation of the von Bezold-Jarisch reflex in anesthetized rats

Activation of 5-hydroxytryptamine (5-HT) 5-HT1A, 5-HT2C, 5-HT3, and 5-HT7 receptors modulates the excitability of cardiac vagal motoneurones, but the precise role of 5-HT2A/2B receptors in these phenomena is unclear. We report here the effects of intracisternal (ic) administration of selective 5-HT2A/2B antagonists on the vagal bradycardia elicited by activation of the von Bezold-Jarisch reflex with phenylbiguanide. The experiments were performed on urethane-anesthetized male Wistar rats (250-270 g, N = 7-9 per group). The animals were placed in a stereotaxic frame and their atlanto-occipital membrane was exposed to allow ic injections. The rats received atenolol (1 mg/kg, iv) to block the sympathetic component of the reflex bradycardia; 20-min later, the cardiopulmonary reflex was induced with phenylbiguanide (15 µg/kg, iv) injected at 15-min intervals until 3 similar bradycardias were obtained. Ten minutes after the last pre-drug bradycardia, R-96544 (a 5-HT2A antagonist; 0.1 µmol/kg), SB-204741 (a 5-HT2B antagonist; 0.1 µmol/kg) or vehicle was injected ic. The subsequent iv injections of phenylbiguanide were administered 5, 20, 35, and 50 min after the ic injection. The selective 5-HT2A receptor antagonism attenuated the vagal bradycardia and hypotension, with maximal effect at 35 min after the antagonist (pre-drug = -200 ± 11 bpm and -42 ± 3 mmHg; at 35 min = -84 ± 10 bpm and -33 ± 2 mmHg; P < 0.05). Neither the 5-HT2B receptor antagonists nor the vehicle changed the reflex. These data suggest that central 5-HT2A receptors modulate the central pathways of the parasympathetic component of the von Bezold-Jarisch reflex.

Memoranda (3 pages, handwritten) with orders to officers of the Royal Quebec Volunteers

Memoranda (3 pages, handwritten) with orders to officers of the Royal Quebec Volunteers regarding their duties at Quebec. This is unsigned, n.d.