Host-parasite-environment relationship, morphology and molecular analyses of Henneguya eirasi n. sp parasite of two wild Pseudoplatystoma spp. in Pantanal Wetland, Brazil

A new myxosporean species, Henneguya eirasi n. sp., is described parasitizing the gill filaments of Pseudoplatystoma corruscans and Pseudoplatystoma fasciatum (Siluriformes: Pimelodidae) caught in the Patanal Wetland of the state of Mato Grosso, Brazil. The parasite formed white, elongated plasmodia measuring up to 3 mm. Mature spores were ellipsoidal in the frontal view, measuring 37.1 +/- 1.8 mu m in total length, 12.9 +/- 0.8 mu m in body length, 3.4 +/- 0.3 mu m in width, 3.1 +/- 0.1 mu m in thickness and 24.6 +/- 2.2 mu m in the caudal process. Polar capsules were elongated and equal in size, measuring 5.4 +/- 0.5 mu m in length and 0.7 +/- 0.1 mu m in width. Polar filaments had 12-13 coils. Histopathological analysis revealed that the parasite developed in the sub-epithelial connective tissue of the gill filaments and the plasmodia were surrounded by a capsule of host connective tissue. The plasmodia caused slight compression of the adjacent tissues, but no inflammatory response was observed in the infection site. Ultrastructure analysis revealed a single plasmodial wall connected to the ectoplasmic zone through numerous pinocytotic canals. The plasmodial wall exhibited numerous projections and slightly electron-dense material was found in the ectoplasm next to the plasmodial wall, forming a line just below the wall. Partial sequencing of the 18S rDNA gene of H. eirasi n. sp. obtained from P. fasciatum resulted in a total of 1066 bp and this sequence did not match any of the Myxozoa available in the GenBank. Phylogenetic analysis revealed the Henneguya species clustering into clades following the order and family of the host fishes. H. eirasi n. sp. clustered alone in one clade, which was the basal unit for the clade composed of Henneguya species parasites of siluriform ictalurids. The prevalence of the parasite was 17.1% in both fish species examined. Parasite prevalence was not influenced by season, host sex or host size. (C) 2011 Elsevier B.V. All rights reserved.

Light and scanning electron microscopy of Henneguya arapaima n. sp (Myxozoa: Myxobolidae) and histology of infected sites in pirarucu (Arapaima gigas: Pisces: Arapaimidae) from the Araguaia River, Brazil

In this report, we describe Henneguya arapaima n. sp., a parasite of the gill arch and gall bladder of Arapaima gigas (pirarucu) collected in the Araguaia River, in the municipality of Nova Crixas, Goias State, central Brazil. The plasmodia were white, round or ellipsoidal and measured 200-600 mu m. Parasite development was asynchronous and the mature spores were fusifonn and had smooth wall. The spores measurements were (range, with means +/- S.D. in parentheses): total length-48.4-53.1 mu m (51.6 +/- 3.4 mu m), body length-13.5-15.2 mu m (14.2 +/- 0.8 mu m), body width-5.1-6.1 mu m (5.7 +/- 0.5 mu m), body thickness-4.7-5.3 mu m (4.9 +/- 0.2 mu m) and caudal process length-38.0-41.2 mu m (38.3 +/- 2.9 mu m). The polar capsules were elongated and of unequal size, with lengths of 6.3-6.8 mu m (6.5 +/- 0.2) and 6.2-6.6 mu m (6.3 +/- 0.1) for the longest and shortest axes, respectively. Capsule width was 1.4-1.6 mu m (1.5 +/- 0.1). Histological analysis showed that the plasmodia occurred in the tunica adventitia of the gall bladder and were delimited by a thin capsule of connective tissue. In the gill arch, the plasmodia were also surrounded by connective tissue similar to the endomesium, of striated skeletal muscle cells. Sixty-five juvenile specimens of A. gigas weighing 1.0-25.0 kg were examined, 17 (26.1%) of which were infected. Of these, 14 (82.3%) had cysts in the gall bladder, two (11.7%) had cysts in the gill arch and only one (5.9%) had cysts in both organs. When the fish were grouped by weight, the prevalence of infection in fish weighing up to 10.0 kg (20.7%) was significantly lower than in fish weighing 10.1-25.0 kg (50%) (G = 3.93; d.f. = 1; p < 0.05). (C) 2008 Elsevier B.V. All rights reserved.

Age-dependent changes in adenosine A(1) receptor distribution and density within the nucleus tractus solitarii of normotensive and hypertensive rats

This study shows the distribution and density of adenosine A1 receptor (A(1)R) within the nucleus tractus solitarii (NTS) of Wistar Kyoto (WKY) and spontaneously hypertensive rats (SHR) from birth to adulthood (1, 15, 30 and 90 days old). The NTS shows heterogeneous distribution of A(1)R in dorsomedial/dorsolateral, subpostremal and medial/intermediate subnuclei. A(1)R decrease from rostral to caudal within dorsomedial/dorsolateral subnucleus in 15-, 30- and 90-day-old WKY and SHR. A(1)R increase from rostral to caudal subpostremal subnucleus in 30- and 90-day-old WKY, and in 15-, 30- and 90-day-old SHR. Furthermore, A(1)Rs are increased in SHR as compared with WKY within dorsomedial/dorsolateral in 30- and 90-day-old and within subpostremal of 15-, 30- and 90-day-old rats. Finally, A(1)Rs increase from 1- to 30-day-old rats. Medial/intermediate did not show any changes in A(1)R from rostral to caudal levels, age or strain. In summary, our result highlights the importance of A1 adenosine system regarding the neural control of blood pressure and the development of hypertension.

Human Multipotent Mesenchymal Stromal Cells from Distinct Sources Show Different In Vivo Potential to Differentiate into Muscle Cells When Injected in Dystrophic Mice

Limb-girdle muscular dystrophies are a heterogeneous group of disorders characterized by progressive degeneration of skeletal muscle caused by the absence or deficiency of muscle proteins. The murine model of Limb-Girdle Muscular Dystrophy 2B, the SJL mice, carries a deletion in the dysferlin gene. Functionally, this mouse model shows discrete muscle weakness, starting at the age of 4-6 weeks. The possibility to restore the expression of the defective protein and improve muscular performance by cell therapy is a promising approach for the future treatment of progressive muscular dystrophies (PMD). We and others have recently shown that human adipose multipotent mesenchymal stromal cells (hASCs) can differentiate into skeletal muscle when in contact with dystrophic muscle cells in vitro and in vivo. Umbilical cord tissue and adipose tissue are known rich sources of multipotent mesenchymal stromal cells (MSCs), widely used for cell-based therapy studies. The main objective of the present study is to evaluate if MSCs from these two different sources have the same potential to reach and differentiate in muscle cells in vivo or if this capability is influenced by the niche from where they were obtained. In order to address this question we injected human derived umbilical cord tissue MSCs (hUCT MSCs) into the caudal vein of SJL mice with the same protocol previously used for hASCs; we evaluated the ability of these cells to engraft into recipient dystrophic muscle after systemic delivery, to express human muscle proteins in the dystrophic host and their effect in functional performance. These results are of great interest for future therapeutic application.

Morphology and phylogenetic relationships of a remarkable new genus and two new species of Neotropical freshwater stingrays from the Amazon basin (Chondrichthyes: Potamotrygonidae)

The morphology and phylogenetic relationships of a new genus and two new species of Neotropical freshwater stingrays, family Potamotrygonidae, are investigated and described in detail. The new genus, Heliotrygon, n. gen., and its two new species, Heliotrygon gomesi, n. sp. (type-species) and Heliotrygon rosai, n. sp., are compared to all genera and species of potamotrygonids, based on revisions in progress. Some of the derived features of Heliotrygon include its unique disc proportions (disc highly circular, convex anteriorly at snout region, its width and length very similar), extreme subdivision of suborbital canal (forming a complex honeycomb-like pattern anterolaterally on disc), stout and triangular pelvic girdle, extremely reduced caudal sting, basibranchial copula with very slender and acute anterior extension, and precerebral and frontoparietal fontanellae of about equal width, tapering very little posteriorly. Both new species can be distinguished by their unique color patterns: Heliotrygon gomesi is uniform gray to light tan or brownish dorsally, without distinct patterns, whereas Heliotrygon rosai is characterized by numerous white to creamy-white vermiculate markings over a light brown, tan or gray background color. Additional proportional characters that may further distinguish both species are also discussed. Morphological descriptions are provided for dermal denticles, ventral lateral-line canals, skeleton, and cranial, hyoid and mandibular muscles of Heliotrygon, which clearly corroborate it as the sister group of Paratrygon. Both genera share numerous derived features of the ventral lateral-line canals, neurocranium, scapulocoracoid, pectoral basals, clasper morphology, and specific patterns of the adductor mandibulae and spiracularis medialis muscles. Potamotrygon and Plesiotrygon are demonstrated to share derived characters of their ventral lateral-line canals, in addition to the presence of angular cartilages. Our morphological phylogeny is further corroborated by a molecular phylogenetic analysis of cytochrome b based on four sequences (637 base pairs in length), representing two distinct haplotypes for Heliotrygon gomesi. Parsimony analysis produced a single most parsimonious tree revealing Heliotrygon and Paratrygon as sister taxa (boot-strap proportion of 70%), which together are the sister group to a clade including Plesiotrygon and species of Potamotrygon. These unusual stingrays highlight that potamotrygonid diversity, both in terms of species composition and undetected morphological and molecular patterns, is still poorly known.

Two new species of Loricariidae (Teleostei: Siluriformes) from main channels of the upper and middle Amazon Basin, with discussion of deep water specialization in loricariids

Hemiancistrus pankimpuju, new species, and Panaque bathyphilus, new species, are described from the main channel of the upper (Maranon) and middle (Solimoes)Amazon River, respectively. Both species are diagnosed by having a nearly white body, long filamentous extensions of both simple caudal-fin rays, small eyes, absence of an iris operculum and unique combinations of morphometrics. The coloration and morphology of these species, unique within Loricariidae, are hypothesized to be apomorphies associated with life in the dark, turbid depths of the Amazon mainstem. Extreme elongation of the caudal filaments in these and a variety of other main channel catfishes is hypothesized to have a mechanosensory function associated with predator detection.

Moenkhausia celibela: a new species from the Amazon basin, Brazil (Characiformes: Characidae)

A new species of Characidae, Moenkhausia celibela, is described from the Rio Amazonas at Santarem, Rio Marau, several localities in the Rio Tapajos, Rio Curua-Una, Rio Xingu and Rio Jari, all from the Amazon basin, Brazil. The new species is distinguished from its congeners, except species included in Gery`s 1992 Moenkhausia lepidura group, by presenting a dark blotch on the upper caudal-fin lobe, and the lower lobe is hyaline or light grey. Moenkhausia celibela is distinguished from the species of the M. lepidura group by the absence of a humeral spot and the presence of a roughly triangular and dark spot at the caudal-fin base, extending posteriorly along the middle caudal-fin rays, and distinctly separate from the spot on the upper caudal-fin lobe.

A new species of Moenkhausia from the rio Amazonas and rio Orinoco basins (Characiformes: Characidae)

A new species of Moenkhausia is described from the rio Amazonas and rio Orinoco basins. The new species can be distinguished from congeners mainly by the combination of a conspicuous, relatively small and circular humeral spot, a black spot on the upper caudal-fin lobe, lower caudal-fin lobe without spot or a faint one, and middle caudal-fin rays hyaline or with dark tips. Mature males have a unique combination of two large-sized bony hooks on the anal-fin rays and tiny spines on the distal portion of all fins, which distinguishes the new species from any other species of Characidae.

Systematics and phylogeny of Cristacoxidae (Copepoda, Harpacticoida): a review

Both sexes of a new species of Noodtorthopsyllus Lang, 1965 (Harpacticoida, Cristacoxidae) from a sandy beach in Sao Paulo State (Brazil) are described using light and scanning electron microscopy. Noodtorthopsyllus tageae sp. nov. displays a mosaic of characters drawn from both Noodtorthopsyllus and Cristacoxa Huys, 1990, blurring the boundaries between both genera. Consequently, Cristacoxa, the type genus of the nominal family-group taxon Cristacoxidae Huys, 1990, is relegated to a junior subjective synonym of Noodtorthopsyllus, and its type species is transferred to the latter as N. petkovskii (Huys, 1990) comb. nov. A new genus Acuticoxa is proposed to accommodate A. ubatubaensis sp. nov. (type species), collected on the northern continental shelf of Sao Paulo State, and A. biarticulata sp. nov., previously identified as Laophontisochra sp., from the Northern Magellan Straits. Amended diagnoses are provided for Noodtorthopsyllus and Laophontisochra. Autapomorphies supporting the monophyly of the Cristacoxidae are re-evaluated, including new data on P3 endopod sexual dimorphism and caudal ramus development. It is concluded that a recently published hypothesis of a deeply rooted split of the family into two highly divergent lineages cannot be supported. Consequently, both Laophontisochra and Acuticoxa gen. nov. are removed from the Cristacoxidae and tentatively assigned to the Nannopodidae (ex Huntemanniidae), forming a clade with three other genera displaying coxal modifications on leg 1 (Rosacletodes Wells, 1985; Huntemannia Poppe, 1884; and an as yet undescribed genus from Brazil). Based on the sexual dimorphism of the P4 endopod, we propose to transfer Metahuntemannia Smirnov, 1946 and Pottekia Huys, 2009 from the Nannopodidae to the Canthocamptidae (subfamily Hemimesochrinae) where they are probably most closely related to Psammocamptus Mielke, 1975; Bathycamptus Huys & Thistle, 1989; Perucamptus Huys & Thistle, 1989; and Isthmiocaris George & Schminke, 2003. An identification key to the genera of the Nannopodidae is presented.

Contributions to the taxonomy of the Normanellidae (Copepoda, Harpacticoida): description of a new genus from the Brazilian continental shelf and re-assignment of Pseudocletodes vararensis Scott & Scott, 1893 (ex Nannopodidae)

A new genus and species of Normanellidae (Copepoda, Harpacticoida), Paranaiara inajae gen. et sp. nov., is described from the continental shelf off the northern coast of Sao Paulo State, Brazil. The new genus differs from the type genus Normanella Brady, 1880 and Sagamiella Lee & Huys, 1999 in its presence of lamelliform caudal rami, a maxillulary endopod represented by 2 setae, an unarmed maxillipedal syncoxa, and reduced setation on P2 enp-2 (without outer spine) and P3 enp-2 (with only 2 inner setae). All these apomorphic character states are shared with the genus Pseudocletodes Scott & Scott, 1893, formerly placed in the family Nannopodidae (ex Huntemanniidae) and here assigned to the Normanellidae. Pseudocletodes can be differentiated from Paranaiara by the loss of the P1 endopod and of the inner seta on P2-P4 enp-1, the presence of only 2 inner setae on P2 enp-2 (instead of 3) and only 1 inner seta on P4 exp-3 (instead of 2), the presence of a second inner seta on P4 enp-2 (instead of 1), the morphology of the fifth pair of legs which are not medially fused and have only 3 endopodal elements (instead of 4) in the male, and the well developed caudal ramus seta V (instead of rudimentary). It is postulated that prehensility of the P1 endopod was secondarily lost in the common ancestor of Paranaiara and Pseudocletodes. An updated family diagnosis of the Normanellidae and a dichotomous identification key to the 22 currently valid species are presented.

Central mechanisms involved in pilocarpine-induced pressor response

Pilocarpine (cholinergic muscarinic agonist) injected peripherally may act centrally to produce pressor responses; in the present study, using c-fos immunoreactive expression, we investigated the forebrain and brainstem areas activated by pressor doses of intravenous (i.v.) pilocarpine. In addition, the importance of vasopressin secretion and/or sympathetic activation and the effects of lesions in the anteroventral third ventricle (AV3V) region in awake rats were also investigated. In male Holtzman rats, pilocarpine (0.04 to 4 mu mol/kg b.w.) i.v. induced transitory hypotension followed by long lasting hypertension. Sympathetic blockade with prazosin (1 mg/kg b.w.) i.v. or AV3V lesions (1 day) almost abolished the pressor response to i. v. pilocarpine (2 mu mol/kg b.w.), whereas the vasopressin antagonist (10 mu g/kg b.w.) i.v. reduced the response to pilocarpine. Pilocarpine (2 and 4 mu mol/kg b.w.) i.v. increased the number of c-fos immunoreactive cells in the subfornical organ, paraventricular and supraoptic nuclei of the hypothalamus, organ vasculosum of the lamina terminalis, median preoptic nucleus, nucleus of the solitary tract and caudal and rostral ventrolateral medulla. These data suggest that i.v. pilocarpine activates specific forebrain and brainstem mechanisms increasing sympathetic activity and vasopressin secretion to induce pressor response. (C) 2011 Elsevier B.V. All rights reserved.

Morphometric analysis of the AMPA-type neurons in the Deiters vestibular complex of the chick brain

Chicken (Gallus gallus) brains were used to investigate the typology and the immunolabel pattern for the subunits composing the AMPA-type glutamate receptors (GluR) of hindbrain neurons of the dorsal (dND) and ventral nuclei (vND) of the Deiter`s vestibular complex (CD), which is the avian correspondent of the lateral vestibular nucleus (LVN) of mammals. Our results revealed that neurons of both divisions were poor in GluR1. The vND, the GluR2/3+ and GluR4+ label presented no area or neuronal size preference, although most neurons were around 75%. The dND neurons expressing GluR2/3 are primarily around 85%, medium to large-sized 85%, and predominantly 60% located in the medial portion of the rostral pole and in the lateral portion of the caudal pole. The majority of dND neurons containing GluR4 are also around 75%, larger (70% are large and giant), exhibiting a distribution that seems to be complementary to that of GluR2/3+ neurons. This distinct arrangement indicates functional differences into and between the DC nuclei, also signaling that such variation could be attributed to the diverse nature of the subunit composition of the GluRs. Discussion addresses the morphological and functional correlation of the avian DC with the LVN of mammals in addition to the high morphological correspondence, To include these data into the modern comparative approach we propose to adopt a similar nomenclature for the avian divisions dND and vND that could be referred as dLVN and vLVN. (C) 2008 Elsevier B.V. All rights reserved.

Prefrontal afferents to the dorsal raphe nucleus in the rat

The prefrontal cortex (PFC) receives strong inputs from monoaminergic cell groups in the brainstem and also sends projections to these nuclei. Recent evidence suggests that the PFC exerts a powerful top-down control over the dorsal raphe nucleus (DR) and that it may be involved in the actions of pharmaceutical drugs and drugs of abuse. In the light of these findings, the precise origin of prefrontal inputs to DR was presently investigated by using the cholera toxin subunit b (CTb) as retrograde tracer. All the injections placed in DR produced retrograde labeling in the medial, orbital, and lateral divisions of the PFC as well as in the medial part of the frontal polar cortex. The labeling was primarily located in layer V. Remarkably, labeling in the medial PFC was denser in its ventral part (infralimbic and ventral prelimbic cortices) than in its dorsal part (dorsal prelimbic, anterior cingulate and medial precentral cortices). After injections in the rostral or caudal DR, the largest number of labeled neurons was observed in the medial PFC, whereas after injections in the mid-rostrocaudal DR, the labeled neurons were more homogeneously distributed in the three main PFC divisions. A cluster of labeled neurons also was observed around the apex of the rostral pole of the accumbens, especially after rostral and mid-rostrocaudal DR injections. Overall, these results confirm the existence of robust preftontal projections to DR, mainly derived from the ventral part of the medial PFC, and underscore a substantial contribution of the frontal polar cortex. (C) 2008 Elsevier Inc. All rights reserved.

Organization of ventral tegmental area projections to the ventral tegmental area-nigral complex in the rat

The ventral tegmental area (VTA) is a nodal link in reward circuitry. Based on its striatal output, it has been subdivided in a caudomedial part which targets the ventromedial striatum, and a lateral part which targets the ventrolateral striatum [Ikemoto S (2007) Dopamine reward circuitry: two projection systems from the ventral midbrain to the nucleus accumbens-olfactory tubercle complex. Brain Res Rev 56:27-78]. Whether these two VTA parts are interconnected and to what extent the VTA innervates the substantia nigra compacta (SNc) and retrorubral nucleus (RR) are critical issues for understanding information processing in the basal ganglia. Here, VTA projections to the VTA-nigral complex were examined in rats, using Phaseolus vulgaris leucoagglutinin (PHA-L) as anterograde tracer. The results show that the dorsolateral VTA projects to itself, as well as to the dorsal tier of the SNc and RR, largely avoiding the caudomedial VTA. The ventrolateral VTA innervates mainly the interfascicular nucleus. The components of the caudomedial VTA (the interfascicular, paranigral and caudal linear nuclei) are connected with each other. In addition, the caudomedial VTA (especially the paranigral and caudal linear nuclei) innervates the lateral VTA, and, to a lesser degree, the SNc and RR. The caudal pole of the VTA sends robust, bilateral projections to virtually all the VTA-nigral complex, which terminate in the dorsal and ventral tiers. Modest inputs from the medial supramammillary nucleus to ventromedial parts of the VTA-nigral complex were also identified. In double-immunostained sections, PHA-L-labeled varicosities were sometimes found apposed to tyrosine hydroxylase-positive neurons in the ventral mesencephalon. Overall, the results underscore that VTA projections to the VTA-nigral complex are substantial and topically organized. In general, these projections, like the spiralated striato-nigro-striatal loops, display a medial-to-lateral organization. This anatomical arrangement conceivably permits the ventromedial striatum to influence the activity of the lateral striatum. The caudal pole of the VTA appears to be a critical site for a global recruitment of the mesotelencephalic system. (C) 2008 IBRO. Published by Elsevier Ltd. All rights reserved.

Proposta de ferramenta para validação temporal em barramentos de campo

Sistemas de controle industriais precisam atender a requisitos temporais para garantir seu correto funcionamento, sendo por isto considerados sistemas tempo-real. Quando tais sistemas são distribuídos, tais como redes de sensores, atuadores e controladores interligados por barramentos de campo, a comunicação desempenha um papel importante no comportamento temporal. Este trabalho propõe uma ferramenta para validar o comportamento temporal da comunicação em um protocolo de barramento de campo, o Foundation Fieldbus. A proposta inclui a especificação de requisitos e a visualização da validação. Pretende-se auxiliar a compreensão do comportamento temporal e possibilitar a depuração de sistemas tempo-real distribuídos. O sistema desenvolvido encontra-se operacional e foi validado em diversos estudos de caso, os quais são descritos no presente trabalho.