Genotoxic effect of formocresol pulp therapy of deciduous teeth

OBJECTIVE: To investigate whether formocresol, in Buckley's original formulation, used for pulp therapy of deciduous teeth, can have a genotoxic effect. Genotoxicity was tested in lymphocyte cultures from the peripheral blood of children aged 5-10y, in Recife, Pernambuco, Brazil. This was a case-control study. The sample comprised 40 children who had primary teeth with non-vital pulps. Two venous blood samples (6-8ml) were collected from each child, the first prior to pulp therapy (control group) and the second 24h after pulp therapy (experimental group). Lymphocyte cultures were grown in 78% RPMI 1640 medium, 20% fetal bovine serum, 2% phytohemagglutinin. The lymphocytes were assessed for chromosomal aberrations; each sample involved analysis of 100 metaphases. There was a statistically significant difference between the control and treated groups for the isochromatid gap (p<0.001), chromatid break (p<0.009), isochromatid break (p<0.046), other chromosomal alterations (p<0.001), and for total aberrations. In view of these results, caution in the use of formocresol in pediatric dentistry is recommended.     

Symbiotic association of a pyrgomatid barnacle with a coral from a volcanic middle Miocene shoreline (Porto Santo,Madeira Archipelago,Portugal)

Abstract: The bioclaustration of the pyrgomatid barnacle Ceratoconcha aff. costata within the carbonate skeleton of a colonial hermatypical coral (Tarbellastrea reussiana) is described from a middle Miocene basaltic rocky palaeoshore on a small north‐eastern Atlantic islet of Porto Santo (Madeira Archipelago, Portugal). The resulting structure is named as a new trace fossil Imbutichnus igen. nov., characterized by a small, funnel‐like cavity, a circular to oval cross‐section, with a nearly cylindrical shape in the upper part and a conical shape towards the base. Imbutichnus costatum isp. nov. is defined as a bioclaustration structure produced by the overgrowth of a pyrgomatid barnacle by a coral. From an ethologic point of view, Imbutichnus is attributed to the Impedichnia category. In terms of palaeoecology, it is interpreted as the result of a parasitic relationship. This is also the oldest record of pyrgomatid barnacles in the eastern Atlantic and clearly demonstrates that the Miocene palaeogeographic range of Ceratoconcha was much wider than previously assumed.     

Leishmania (Viannia) braziliensis: insights on subcellular distribution and biochemical properties of heparin-binding proteins

Leishmaniasis is a vector-borne disease and an important public health issue. Glycosaminoglycan ligands in Leishmania parasites are potential targets for new strategies to control this disease. We report the subcellular distribution of heparin-binding proteins (HBPs) in Leishmania (Viannia) braziliensis and specific biochemical characteristics of L. (V.) braziliensis HBPs. Promastigotes were fractionated, and flagella and membrane samples were applied to HiTrap Heparin affinity chromatography columns. Heparin-bound fractions from flagella and membrane samples were designated HBP Ff and HBP Mf, respectively. Fraction HBP Ff presented a higher concentration of HBPs relative to HBP Mf, and SDS-PAGE analyses showed 2 major protein bands in both fractions (65 and 55 kDa). The 65 kDa band showed gelatinolytic activity and was sensitive to inhibition by 1,10-phenanthroline. The localization of HBPs on the promastigote surfaces was confirmed using surface plasmon resonance (SPR) biosensor analysis by binding the parasites to a heparin-coated sensor chip; that was inhibited in a dose-dependent manner by pre-incubating the parasites with variable concentrations of heparin, thus indicating distinct heparin-binding capacities for the two fractions. In conclusion, protein fractions isolated from either the flagella or membranes of L. (V.) braziliensis promastigotes have characteristics of metallo-proteinases and are able to bind to glycosaminoglycans.     

An integrative approach to characterize cryptic species in the Thoracostoma trachygaster Hope, 1967 complex (Nematoda: Leptosomatidae)

Nematode diversity may seriously be underestimated when taking into account cryptic speciation. Thoracostoma trachygaster is commonly found in kelp holdfasts along the California coastline and was recently shown to consist of at least two distinct molecular clades (I and II). Here, we provide detailed morphological analysis of both clades, based on measurements taken from video vouchers of respectively eight and 16 individuals from the previous study, as well as 80 newly collected specimens from four Californian beaches. The latter were vouchered, measured, and then subjected to molecular analyses of the mitochondrial cytochrome oxidase c subunit I (COI) gene, and the ribosomal D2D3 and internal transcribed spacer (ITS) regions. This integrative approach shows that the three molecular clades are phylogenetically and morphologically distinct species, but a combination of morphological characters is needed to distinguish them. Two new species, Thoracostoma fatimae sp. nov. and Thoracostoma igniferum sp. nov. , are identified and described. The spicule length of T. fatimae sp. nov. is significantly shorter than that of T. trachygaster. Thoracostoma igniferum sp. nov. can be distinguished by the irregular posterior edge of the cephalic capsule and the two internal subdorsal tropis‐like projections in the wall of the cephalic capsule, which are lacking in T. fatimae sp. nov. and T. trachygaster. © 2012 The Linnean Society of London, Zoological Journal of the Linnean Society, 2012, 164 , 18–35.     

Glandular trichomes in Connarus suberosus (Connaraceae): distribution, structural organization and probable functions

Connarus suberosus is a typical species of the Brazilian Cerrado biome, and its inflorescences and young vegetative branches are densely covered by dendritic trichomes. The objective of this study was to report the occurrence of a previously undescribed glandular trichome of this species. The localization, origin and structure of these trichomes were investigated under light, transmission and scanning electron microscopy. Collections were made throughout the year, from five adult specimens of Connarus suberosus near Botucatu, S?o Paulo, Brazil, including vegetative and reproductive apices, leaves and fruits in different developmental stages, as well as floral buds and flowers at anthesis. Glandular trichomes (GTs) occurred on vegetative and reproductive organs during their juvenile stages. The GTs consisted of a uniseriate, multicellular peduncle, whose cells contain phenolic compounds, as well as a multicellular glandular portion that accumulates lipids. The glandular cell has thin wall, dense cytoplasm (with many mitochondria, plastids and dictyosomes), and a large nucleus with a visible nucleolus. The starch present in the plastids was hydrolyzed during the synthesis phase, reducing the density of the plastid stroma. Some plastids were fused to vacuoles, and some evidence suggested the conversion of plastids into vacuoles. During the final activity stages of the GTs, a darkening of the protoplasm was observed in some of the glandular cells, as a programmed cell death; afterwards, became caducous. The GTs in C. suberosus had a temporal restriction, being limited to the juvenile phase of the organs. Their presence on the exposed surfaces of developing organs and the chemical nature of the reserve products, suggest that these structures are food bodies. Field observations and detailed studies of plant-environment interactions, as well as chemical analysis of the reserve compounds, are still necessary to confirm the role of these GTs as feeding rewards.     

Non-classic androgen actions in Sertoli cell membrane in whole seminiferous tubules: effects of nandrolone decanoate and catechin

Studies show a mechanism of action of testosterone, nandrolone and catechin as agonists of the membrane androgen receptor. The aim of this work is to investigate the non-classical effect of androgens and catechin in Sertoli cells from immature rats. The membrane potential of Sertoli cells in whole seminiferous tubules was recorded using a standard single microelectrode technique. It was performed a topical application of testosterone (1 μM), nandrolone (0.1, 0.5 and 1 μM) and the flavonoid catechin (0.1, 0.5 and 1 μM) alone and also after infusion with flutamide (1 μM), diazoxide (100 μM) or U73122 (1 μM). The immature testes were incubated for 5 min in KRb with (45)Ca(2+), with or without nandrolone (1 μM). The results were given as mean±SEM. The data were analyzed using ANOVA for repeated measures with Bonferroni post-test. Testosterone produces a depolarization in the membrane potential at 120 s after application. Catechin (1 μM) and nandrolone (1 μM) have shown a similar response to testosterone: depolarization at 120 s after the application. The same response of catechin and nandrolone was observed at different doses. The effects of testosterone, catechin and nandrolone were not affected after perfusion with flutamide. Perfusion with diazoxide and U73122 nullified the effect of nandrolone (1 μM) and catechin (1 μM). Nandrolone and testosterone increased (45)Ca(2+) uptake with or without flutamide within 5min. These results indicate that nandrolone and catechin act through a receptor on the plasmatic membrane, as well as testosterone, showing a non-classical pathway in Sertoli cells from immature rat testes.     

Antibodies against the envelope glycoprotein promote infectivity of immature dengue virus serotype 2

Cross-reactive dengue virus (DENV) antibodies directed against the envelope (E) and precursor membrane (prM) proteins are believed to contribute to the development of severe dengue disease by facilitating antibody-dependent enhancement of infection. We and others recently demonstrated that anti-prM antibodies render essentially non-infectious immature DENV infectious in Fcγ-receptor-expressing cells. Immature DENV particles are abundantly present in standard (st) virus preparations due to inefficient processing of prM to M during virus maturation. Structural analysis has revealed that the E protein is exposed in immature particles and this prompted us to investigate whether antibodies to E render immature particles infectious. To this end, we analyzed the enhancing properties of 27 anti-E antibodies directed against distinct structural domains. Of these, 23 bound to immature particles, and 15 enhanced infectivity of immature DENV in a furin-dependent manner. The significance of these findings was subsequently tested in vivo using the well-established West Nile virus (WNV) mouse model. Remarkably, mice injected with immature WNV opsonized with anti-E mAbs or immune serum produced a lethal infection in a dose-dependent manner, whereas in the absence of antibody immature WNV virions caused no morbidity or mortality. Furthermore, enhancement infection studies with standard (st) DENV preparations opsonized with anti-E mAbs in the presence or absence of furin inhibitor revealed that prM-containing particles present within st virus preparations contribute to antibody-dependent enhancement of infection. Taken together, our results support the notion that antibodies against the structural proteins prM and E both can promote pathogenesis by enhancing infectivity of prM-containing immature and partially mature flavivirus particles.