Bristle pattern formation in the thorax formation of Drosophila: a systems level approach

A fundamental question in developmental biology is how tissues are patterned to give rise to differentiated body structures with distinct morphologies. The Drosophila wing disc offers an accessible model to understand epithelial spatial patterning. It has been studied extensively using genetic and molecular approaches. Bristle patterns on the thorax, which arise from the medial part of the wing disc, are a classical model of pattern formation, dependent on a pre-pattern of trans-activators and –repressors. Despite of decades of molecular studies, we still only know a subset of the factors that determine the pre-pattern. We are applying a novel and interdisciplinary approach to predict regulatory interactions in this system. It is based on the description of expression patterns by simple logical relations (addition, subtraction, intersection and union) between simple shapes (graphical primitives). Similarities and relations between primitives have been shown to be predictive of regulatory relationships between the corresponding regulatory factors in other Systems, such as the Drosophila egg. Furthermore, they provide the basis for dynamical models of the bristle-patterning network, which enable us to make even more detailed predictions on gene regulation and expression dynamics. We have obtained a data-set of wing disc expression patterns which we are now processing to obtain average expression patterns for each gene. Through triangulation of the images we can transform the expression patterns into vectors which can easily be analysed by Standard clustering methods. These analyses will allow us to identify primitives and regulatory interactions. We expect to identify new regulatory interactions and to understand the basic Dynamics of the regulatory network responsible for thorax patterning. These results will provide us with a better understanding of the rules governing gene regulatory networks in general, and provide the basis for future studies of the evolution of the thorax-patterning network in particular.

Antithrombotic effect of Lonomia obliqua caterpillar bristle extract on experimental venous thrombosis

The venom of Lonomia obliqua caterpillar may induce a hemorrhagic syndrome in humans, and blood incoagulability by afibrinogenemia when intravenously injected in laboratory animals. The possible antithrombotic and thrombolytic activities of L. obliqua caterpillar bristle extract (LOCBE) were evaluated in this study. The minimal intravenous dose of the extract necessary to induce afibrinogenemia and anticoagulation was 3.0 and 10.0 µg protein/kg body weight for rabbits and rats, respectively. In rabbits, this dose induced total blood incoagulability for at least 10 h and did not reduce the weight of preformed venous thrombi, in contrast to streptokinase (30,000 IU/kg). In rats, pretreatment with 5.0 and 10.0 µg/kg LOCBE prevented the formation of thrombi induced by venous stasis or by injury to the venous endothelium. The dose of 5.0 µg/kg LOCBE did not modify blood coagulation assay parameters but increased bleeding time and decreased plasma factor XIII concentration. When the extract was administered to rats at the dose of 10.0 µg/kg, the blood was totally incoagulable for 6 h. These data show that LOCBE was effective in preventing experimental venous thrombosis in rats, justifying further studies using purified fractions of the extract to clarify the mechanisms of this effect.

Lonomia obliqua (Lepidoptera, Saturniidae) caterpillar bristle extract induces direct lysis by cleaving erythrocyte membrane glycoproteins

Lonomia obliqua caterpillar bristle extract induces hemolysis in vitro on washed human and rat erythrocytes, in either the absence or presence of exogenous lecithin. In the former condition, phospholipases A(2) are key enzymes involved in hemolysis. However, the mechanism whereby this extract causes direct hemolysis is not known. Thus, the aim of this study was to investigate the hemolytic mechanism of the crude extract of the caterpillar L obliqua on human erythrocytes in the absence of lecithin. The extract significantly increased the erythrocyte osmotic fragility and promoted the removal of glycophorins A and C, and band 3 from the erythrocyte membrane. The use of Ca(2+) and Mg(2+) ions significantly potentiated glycoprotein removal, remarkably of erythrocyte band 3. The composition of fatty acids was analyzed by HPLC in both L obliqua caterpillar bristle extract and human erythrocyte membranes incubated with the extract. The levels of unsaturated fatty acids were remarkably augmented in erythrocytes incubated with the extract than in control erythrocytes, modifying thereby the saturated/unsaturated fatty acid ratio. Altogether, evidence is provided here that the interplay of at least three mechanisms of action accounts for the direct activity of the bristle extract on erythrocyte membrane, leading to hemolysis: the removal of glycoproteins and band 3; the insertion of fatty acids; and the action of phospholipases. Such mechanisms might affect erythrocyte flexibility and deformability, which may induce hemolysis by increasing erythrocyte fragility. However, whether the direct hemolytic activity of L obliqua caterpillar is the major cause of intravascular hemolysis during envenomation still needs further investigation. (C) 2010 Elsevier Ltd. All rights reserved.

The differential roles of D-Pax2 variants in regulating Drosophila eye and bristle development

The ability to appropriately interact with the environment is crucial to an organism’s survival. The establishment of functional sensory systems, such as the bristles and eyes in Drosophila, is a critical event during the development of the organism. The transcription factor D Pax2 is involved in the differentiation of the shaft and glial cells in the developing bristle (Kavaler et al., Dev, 126:2261-2272, 1999) and of the cone and primary pigment cells in the developing eye (Fu and Noll, Genes Dev, 11:389-405, 1997). How D-Pax2 contributes to distinct differentiative pathways in different cell types is not known. Recent work by Anna Czechowski and Katherine Harmon (personal communication) identified a mutation in the D-Pax2 gene that introduced a stop codon at the end of exon 9, effectively truncating the protein. This mutation affects bristle, but not eye, development. We thus suspected regions after exon 9 are required for D-Pax2 function only in the bristles and may also be associated with alternative splicing of the D Pax2 transcript. We plan to assess the role of the carboxy terminal region of the protein by establishing transgenic lines bearing rescue constructs of D-Pax2 with either the complete coding sequence or with deletions of specific exons. To date, we have generated the first rescue construct bearing the complete coding region of the gene driven by a 3 KB upstream regulatory region of D-Pax2 and are currently generating transgenic fly lines with this construct.

Antithrombotic effect of Lonomia obliqua caterpillar bristle extract on experimental venous thrombosis

The venom of Lonomia obliqua caterpillar may induce a hemorrhagic syndrome in humans, and blood incoagulability by afibrinogenemia when intravenously injected in laboratory animals. The possible antithrombotic and thrombolytic activities of L. obliqua caterpillar bristle extract (LOCBE) were evaluated in this study. The minimal intravenous dose of the extract necessary to induce afibrinogenemia and anticoagulation was 3.0 and 10.0 µg protein/kg body weight for rabbits and rats, respectively. In rabbits, this dose induced total blood incoagulability for at least 10 h and did not reduce the weight of preformed venous thrombi, in contrast to streptokinase (30,000 IU/kg). In rats, pretreatment with 5.0 and 10.0 µg/kg LOCBE prevented the formation of thrombi induced by venous stasis or by injury to the venous endothelium. The dose of 5.0 µg/kg LOCBE did not modify blood coagulation assay parameters but increased bleeding time and decreased plasma factor XIII concentration. When the extract was administered to rats at the dose of 10.0 µg/kg, the blood was totally incoagulable for 6 h. These data show that LOCBE was effective in preventing experimental venous thrombosis in rats, justifying further studies using purified fractions of the extract to clarify the mechanisms of this effect.

Mechanical removal of necrotic periodontal ligament by either Robinson bristle brush with pumice or scalpel blade. Histomorphometric analysis and scanning electron microscopy

One of the important factors accounting for successful delayed replantation of avulsed teeth is seemingly the type of root surface treatment. Removal of necrotic cemental periodontal ligament remnants may prevent the occurrence of external root resorption, which is the major cause of loss of teeth replanted in such conditions. The purpose of this study was to compare the efficacy of two mechanical techniques for removal of root-adhered periodontal ligament. Preservation or removal of the cementum layer concomitantly with these procedures was also assessed. Forty-five roots of healthy premolars extracted for orthodontic purposes were selected. After extraction, the teeth were kept dry at room temperature for 1 h and then immersed in saline for rehydration for an additional 10 min. Thereafter, the roots were assigned to three groups, as follows: group 1 (control) - the cemental periodontal ligament was preserved; group 2 - removal of the periodontal ligament by scraping root surface with a scalpel blade (SBS); group 3 - periodontal ligament remnants were removed using a Robinson bristle brush at low-speed with pumice/water slurry (RBP). The specimens were analysed histomorphometrically and examined by scanning electron microscopy. The quantitative and qualitative analyses of the results showed that the RBP technique was significantly more effective than the SBS technique for removal of the periodontal ligament remnants adhered to root surface. Both techniques preserved the cementum layer.

Influence of different toothpaste abrasives on the bristle end-rounding quality of toothbrushes

To evaluate the influence of different toothpaste abrasives on the bristle wear and bristle tip morphology of toothbrushes with different degrees of hardness. Material and methods Ninety samples of bovine incisor teeth were used in this study. The samples were randomly divided into three groups according to the bristle hardness of the toothbrush used: soft bristles (S); extra-soft bristles (ES); hard bristles (H). The toothbrushes of each group were randomly divided into six subgroups with five toothbrushes each, according to the abrasive of the toothpaste used in the simulation: Negative control (distilled water); toothpaste 1 (silica); toothpaste 2 (hydrated silica); toothpaste 3 (calcium carbonate, calcium bicarbonate and silica); toothpaste 4 (tetrapotassium pyrophosphate, silica and titanium dioxide); toothpaste 5 (calcium carbonate). The samples were placed in a toothbrushing simulating machine that simulating three months of brushing. The toothbrush bristles were evaluated by the bristle wear index, and the bristle tips morphology was evaluated by the bristle tip morphology index. Results The ES brush presented the highest bristle wear among the toothbrushes. Additionally, the S brushes showed better morphology of the bristles followed by ES and H brushes. The type of abrasive only influenced the bristle tip morphology of the ES brushes. The toothpaste 3 induced the worse bristle tip morphology than all the other toothpastes. Conclusions Different abrasives have influence only on the bristle tip morphology of the ES brushes.

Influence of different toothpaste abrasives on the bristle end-rounding quality of toothbrushes

ObjectivesTo evaluate the influence of different toothpaste abrasives on the bristle wear and bristle tip morphology of toothbrushes with different degrees of hardness.Material and methodsNinety samples of bovine incisor teeth were used in this study. The samples were randomly divided into three groups according to the bristle hardness of the toothbrush used: soft bristles (S); extra-soft bristles (ES); hard bristles (H). The toothbrushes of each group were randomly divided into six subgroups with five toothbrushes each, according to the abrasive of the toothpaste used in the simulation: Negative control (distilled water); toothpaste 1 (silica); toothpaste 2 (hydrated silica); toothpaste 3 (calcium carbonate, calcium bicarbonate and silica); toothpaste 4 (tetrapotassium pyrophosphate, silica and titanium dioxide); toothpaste 5 (calcium carbonate). The samples were placed in a toothbrushing simulating machine that simulating three months of brushing. The toothbrush bristles were evaluated by the bristle wear index, and the bristle tips morphology was evaluated by the bristle tip morphology index.ResultsThe ES brush presented the highest bristle wear among the toothbrushes. Additionally, the S brushes showed better morphology of the bristles followed by ES and H brushes. The type of abrasive only influenced the bristle tip morphology of the ES brushes. The toothpaste 3 induced the worse bristle tip morphology than all the other toothpastes.ConclusionsDifferent abrasives have influence only on the bristle tip morphology of the ES brushes.

Assessment in vitro of brushing on dental surface roughness alteration by laser interferometry

Noncarious cervical lesions (NCCLs) are considered to be of multifactorial origin, normally associated with inadequate brushing. This study assessed the influence in vitro of simulated brushing on NCCL formation. Fifteen human premolars were submitted to brushing in the cementoenamel junction region, using hard-, medium- and soft-bristled brushes associated with a toothpaste of medium abrasiveness under a 200 g load, at a speed of 356 rpm for 100 minutes. The surface topography of the region was analyzed before and after brushing, by means of a laser interferometer, using "cut-off" values of 0.25 and considering roughness values in mm. The initial roughness (mm) results for dentin (D / bristle consistency: 1 - soft, 2 - medium and 3 - hard) were as follows: (D1) 1.25 ± 0.45; (D2) 1.12 ± 0.44; (D3) 1.05 ± 0.41. For enamel (E / bristle consistency: 1 - soft, 2 - medium and 3 - hard), the initial results were: (E1) 1.18 ± 0.35; (E2) 1.32 ± 0.25; (E3) 1.50 ± 0.38. After brushing, the following were the values for dentin: (D1) 2.32 ± 1.99; (D2) 3.30 ± 0.96; (D3) Over 500. For enamel, the values after brushing were: (E1) 1.37 ± 0.31; (E2) 2.15 ± 0.90; (E3) 1.22 ± 0.47. Based on the results of the ANOVA and Tukey statistical analyses (a = .05) it was concluded that soft, medium and hard brushes are not capable of abrading enamel, whereas dentin showed changes in surface roughness by the action of medium- and hard-bristled brushes.

Review of American fossil phlebotominae (Diptera: Psychodidae) with a description of two new species

The objective of this study was to carry out a taxonomic review of fossil American phlebotomine sand flies and describe two new species found in amber in the Dominican Republic. The gonostyle of one of these, Micropygomyia dorafeliciangeliae nov. sp., (=Lutzomyia dorafeliciangeliae, species group oswaldoi), has five spines, similar to that of Micropygomyia paterna (Quate, 1963) (= Lutzomyia paterna, species group oswaldoi), but they may be distinguished by the alpha/gamma ratio, which is <1.0 in the new species and >1 in the latter. Pintomyia dominicana nov. sp. (=Lutzomyia dominicana, species group verrucarum) has four spines on the gonostyle and presents a long bristle on the apex of the paramere, which distinguishes it from the other fossil species. With the description of these two new species, a total of 14 species of the American fossil phlebotomine sand flies has been described, 10 of which belong to the genus Pintomyia. An identification key for male fossil species is presented

The taxonomic status of the endangered thin-spined porcupine, Chaetomys subspinosus (Olfers, 1818), based on molecular and karyologic data

Background: The thin-spined porcupine, also known as the bristle-spined rat, Chaetomys subspinosus (Olfers, 1818), the only member of its genus, figures among Brazilian endangered species. In addition to being threatened, it is poorly known, and even its taxonomic status at the family level has long been controversial. The genus Chaetomys was originally regarded as a porcupine in the family Erethizontidae, but some authors classified it as a spiny-rat in the family Echimyidae. Although the dispute seems to be settled in favor of the erethizontid advocates, further discussion of its affinities should be based on a phylogenetic framework. In the present study, we used nucleotide-sequence data from the complete mitochondrial cytochrome b gene and karyotypic information to address this issue. Our molecular analyses included one individual of Chaetomys subspinosus from the state of Bahia in northeastern Brazil, and other hystricognaths. Results: All topologies recovered in our molecular phylogenetic analyses strongly supported Chaetomys subspinosus as a sister clade of the erethizontids. Cytogenetically, Chaetomys subspinosus showed 2n = 52 and FN = 76. Although the sexual pair could not be identified, we assumed that the X chromosome is biarmed. The karyotype included 13 large to medium metacentric and submetacentric chromosome pairs, one small subtelocentric pair, and 12 small acrocentric pairs. The subtelocentric pair 14 had a terminal secondary constriction in the short arm, corresponding to the nucleolar organizer region (Ag-NOR), similar to the erethizontid Sphiggurus villosus, 2n = 42 and FN = 76, and different from the echimyids, in which the secondary constriction is interstitial. Conclusion: Both molecular phylogenies and karyotypical evidence indicated that Chaetomys is closely related to the Erethizontidae rather than to the Echimyidae, although in a basal position relative to the rest of the Erethizontidae. The high levels of molecular and morphological divergence suggest that Chaetomys belongs to an early radiation of the Erethizontidae that may have occurred in the Early Miocene, and should be assigned to its own subfamily, the Chaetomyinae.

7ª Contribuição ao estudo dos Flebotomus (Diptera: Psychodidae): descrição dos machos de 24 novas espécies

The A. and his co-workers captured in trips in the hinterland of Brazil more tham 17.000 flebotomi from which 35 are new ones, 11 discribed by, him in previous papers. The A. found these insects in groups of species living in different habitats, some ones of them not yet known: ondoors, or outdoors attracted by light or animal baits, without Shannon’s trap, in great or small caves, in the jungle in tree’s holes, holes in stones, holes in the soil habited by animals like armadillos, pacas (Aguti paca), wild rats, cururú toad (Bufo sp.). He observed the life history of 13 species: Flebotomus longipalpis Lutz& Neiva, 1912, Flebotomus intermedius Lutz & Neiva, 1912, Flebotomus avellari Costa Lima, 1932, Flebotomus aragãoi costa Lima, 1932, Flebotomus lutzianus Costa Lima, 1932, Flebotomus limai fonseca, 1935, Flebotomus rickardi Costa Lima, 1936, Flebotomus dasipodogeton Castro, 1939, Flebotomus oswaldoi n. sp., Flebotomus villelai n. sp., Flebotomus triacanthus n. sp., Flebotomus longispinus n. sp. And flebotomus travassosi n. sp. He describes the male of 24 n. sp., explaining the differential diagnose of group or nearly allied species. He inclued F. rooti n. sp. And F. hirsutus n. sp. In the sub-genus Shannonomyia. The first one, very allied to F. davisi Root is different from it, for presenting in the dorsal side of the abdomen bristles and not scales and to have the median claspers longer than his inner appendage and F. hirsutus quite different from the others which show 3 spines on distal segment of the upper clasper and for being the only one who presents the bristles of inner appendage of median clasper longer than it. Only the females of F. amazonensis Root and f. chagasi Costa Lima, are known and then it is possible that they belong to one of the species of this sub-genus from whom only the male have been described. F. choti Floch & Abonnenc, captured also at Pará, F. triacanthus n. sp. F. trispinosus n. sp. And F. equatorialis n. sp. Are very related and to this group the A. proposes the same of Pressatia as sub-genus in honor to whom demonstrated the medical importance of the flebotomi, considering F. triacanthus as the type specie of this sub-genus. In this sub-genus the V papal joint is very long, longer than III + IV, the antennae with geniculated spines without posterior outgrowth. At the genitalia the basal segment of the upper clasper presents two types of bristles ou the inner face, arranged in tuft; the distal segment with 3 spines and 2 thin bristles something difficult to see one of them situated near the apical spine and the other on the base of tubercle where the median spine is articulated; the median clasper is unarmed and compressed; the inferior clasper is also unarmed and longer than de basal segment of the upper clasper; the pompeta is longer than the basal segment of the upper clasper. Following it is presented a key for the determination of the males of the four species of this sub-genus. F. micropygus n. sp., F. minasensis n. sp. e F. dandrophylus n. sp., f. shannoni, F. monticolus, F. pestanai, F. lanei and F. cayenensis constitute a group with many similars characters. F. micropygus is the only American species who present α smaller than β and for that reason and others is allied to. F. minuts and others related species, but presents two terminal spines on the distal segment of the upper clasper. F. micropygus and f. minasensis are quite different because they have very small genitalia, smaller than their heads. F. dendrophylus presents on the median clasper a naked area near the apex and for this and others characters is different from the others of the group. F. flaviscutellatus n. sp., F. oliverioi, F. intermedius and whithmani, are very allied but the first one can be very easily distinguished because it’s scutellum is light. Flebotomus barrettoi n. sp., F. coutinhoi n. sp., F. aragãoi, F. brasiliensis, F. lutzianus, F. texanus, F. pascalei, F. atroclavatus and F. tejeraae are very allied forming a natural group. The two last ones are not well known but the A. A. who have studied them described very long clipeus so long as the head and for that reason can be distinguished from all the others included the two new ones. F. coutinhoi is the only one who presents the apecis of the penis filaments twisted. F. barrettoi n. sp., can be distinguished from aragãoi, texamus and coutinhoi by the length of the penis filaments and from atrocavatus, tejeraae, lutzianus and brasiliensis by the arrangement of the spines of distal segment of the upper clasper. Flebotomus ubiquitalis n. sp., F. auraensis n. sp., F. affinis and F. microps e F. antunesi have many common characters. F. microps n. sp., can be distinguished from any one by the size of the eyes and the presence od well developed genae. This species and other new species are different from F. antunesi by the arrangement of the spines of the distal segment of the upper clasper of the latter. F. ubiquitalis n. sp. can be distinguished from others by the figure of the median clasper. F. auraensis n. sp. Can be distinguished from F. affinis n. sp. By the tuft hairs on the inner face of the basal segment and by arrangement of the spines of the sital segment of the upper clasper. Flebotomus brachipygus n. sp. Seemed to be F. rostrans, specie not well known, by the characters of the genitalia but can not be identified to her by the clypeus size and the palpi’s characters. Flebotomus costalimai, n. sp., f. tupynambai n. sp., and f. castroi Barreto & Coutinho, 1941, are very allied species and the A. proposes to included them the new sub-genus Castromyia, in honor to Dr. G. M. de Oliveira Castro, appointing like typespecies F. castroi with the V joint longer than III + IV; antennae with geniculated spines without posterior prolongation. Genitalia: the basal segment of the upper clasper with a tuft of hairs and the distal segment with 4 spines, one of them at the apex and near it a thin and straight bristle difficult to see; the median clasper with one spinous hair isolated...

Quarta contribuição ao conhecimento do gênero Archytas Jaennicke, 1867: (Diptera, Tachinidae)

The author studies 5 species of Archytas Jaennicke, 1867, belonging to the "dissimilis group": A. seminigra (Wiedemann, 1830) and four species which are considered as new. The species of this group may be characterized as follow: Species of short body, exceptionally large ones. Abdomen yellowish, with a median blackish V-shapedspot. Second antennal segment with 2/3 length of third. Parafacialia with blackish hairs. Propleura pilose. Post alar wall with few hairs. The following key facilitates the identification of the species: 1. Third article of antennae, strongly convex in the anterior margin (fig. 10); posterior margin straight. Parafacialia with a facio-orbital bristle well differentiated . . . . A. arnaudi sp. n. — Third article of antennae not so convex in the anterior margin; facio-orbital bristle absent, if present not well differentiated [...] 2; 2. Parafrontalia with golden polen [...] 3; — Parafrontalia brownish to shining black with few polen . . . 4; 3. Forcipes superiores slender and sub-truncate apically (figs. 5 and 6)[...] A. seminigra; — Forcipes superiores broad apically (fig .20)[...] A, gongalvesi sp. n.; 4. First, second and third sternites yellowish [...] A. angrensis sp. n.; — All sternites brownish to black [...] A. sabroskpi sp. n.; The material studied belongs to the Instituto Oswaldo Cruz collections, where is located the types of new species.

Description of Lutzomyia (Lutzomyia) falquetoi sp. nov. (Diptera: Psychodidae, Phlebotominae) a new species from the state of Espírito Santo, Brazil

Lutzomyia (Lutzomyia) falquetoi, sp. nov. (Diptera: Psychodidae, Phlebotominae) is described from the state of Espírito Santo, Brazil. This new species belongs to the series longipalpis and is easily distinguished from the other members of this taxon by the presence of five well-developed spines and a pre-apical spiniform bristle on the gonostyle as well as nine bristles on the basal tuft of the gonocoxite.