Anomalous somatic embryos in Acca sellowiana (O. Berg) Burret (Myrtaceae)

Somatic embryogenesis represents a valuable tool for the studies on the basic aspects of plant embryo development. Today this process is used as a potencial technique for large-scale plant micropropagation although, so far, it has been applied to only a small number of species. However, when somatic embryos are malformed they are considered economically useless. In Acca sellowiana (O. Berg) Burret, an important fruit-producing crop, large amounts of anomalous somatic embryos (76.3%) were found just after 40 days of culture of explants in a 2,4-D containing medium. Among the anomalous forms found in the cotiledonary stage, 12.2% consisted of fused embryos, 40.4% displayed fused cotyledons, 13.0% presented supernumerary cotyledons, and 10.7% showed absence or poorly developed cotyledons, including those without the shoot apical meristem. Histological analyses indicated that the altered embryos were formed either directly from cotyledons, hypocotyl and radicle of the zygotic embryos used as explants, or indirectly from calli formed from these tissue parts. It is suggested that the formation of anomalous somatic embryos, as well as a low frequency of conversion into emblings reflect physiological and/or genetic disturbances triggered by the presence of 2,4-D in the medium. In vitro experimental alternative approaches are discussed in order to lessen the occurrence of malformed somatic embryos.

Polyamine patterns in haploid and diploid tobacco tissues and in vitro cultures

The aim of this work was to determine PAs levels in pith tissues and callus cultures from haploid and diploid tobacco plants, explanted from the apical and basal regions of the stem. These explants were cultured in an RM-64 medium supplied with IAA and kinetin, under light or in the dark, during successive subcultures. PAs levels followed a basipetal decrease in diploid and an increase in haploid, pith tissues. A similar pattern of total PAs (free + conjugated) was observed for the callus of diploid and haploid plants maintained in the light, and for the haploid callus in the dark, whereas the diploid callus in the dark showed a constant increase in total PAs levels until the end of culture. The PA increase in the diploid callus in the dark was related to free Put levels increase. The ploidy status of the plants could express different PA gradients together with the plant pith and in vitro callus cultures.

Morphoanatomy of the stem in Cyperaceae

Cyperaceae are usually perennial, with underground stems mainly rhizomatous, however, other stem types may also occur, such as corms and tubers. The underground stems of five Cyperaceae species were examined. Cyperus rotundus and Fuirena umbellata have plagiotropic rhizomes, while C. esculentus, C. odoratus, Hypolytrum schraderianum and Bulbostylis paradoxa have orthotropic rhizomes. Corms occur in C. rotundus and C. esculentus, and stolons in C. esculentus. The primary body originates from the activity of the apical meristem and later, from the primary thickening meristem (PTM). Secondary growth results from secondary thickening meristem (STM) activity, and occurs in rhizomes of H. schraderianum, B. paradoxa, C. odotarus and F. umbellata. The procambium and the PTM give rise to collateral bundles in H. schraderianum, and amphivasal bundles in the remaining species. The STM gives rise to the vascular system with the associated phloem and xylem. According to our results, the concept of stem type in Cyperaceae depends on external morphology, function, life phase, activity of the thickening meristems and the relative amount of parenchyma.

Long-term social recognition memory in adult male rats: factor analysis of the social and non-social behaviors

A modified version of the intruder-resident paradigm was used to investigate if social recognition memory lasts at least 24 h. One hundred and forty-six adult male Wistar rats were used. Independent groups of rats were exposed to an intruder for 0.083, 0.5, 2, 24, or 168 h and tested 24 h after the first encounter with the familiar or a different conspecific. Factor analysis was employed to identify associations between behaviors and treatments. Resident rats exhibited a 24-h social recognition memory, as indicated by a 3- to 5-fold decrease in social behaviors in the second encounter with the same conspecific compared to those observed for a different conspecific, when the duration of the first encounter was 2 h or longer. It was possible to distinguish between two different categories of social behaviors and their expression depended on the duration of the first encounter. Sniffing the anogenital area (49.9% of the social behaviors), sniffing the body (17.9%), sniffing the head (3%), and following the conspecific (3.1%), exhibited mostly by resident rats, characterized social investigation and revealed long-term social recognition memory. However, dominance (23.8%) and mild aggression (2.3%), exhibited by both resident and intruders, characterized social agonistic behaviors and were not affected by memory. Differently, sniffing the environment (76.8% of the non-social behaviors) and rearing (14.3%), both exhibited mostly by adult intruder rats, characterized non-social behaviors. Together, these results show that social recognition memory in rats may last at least 24 h after a 2-h or longer exposure to the conspecific.

Molecular markers and genetic diversity of Plasmodium vivax

Enhanced understanding of the transmission dynamics and population genetics for Plasmodium vivax is crucial in predicting the emergence and spread of novel parasite phenotypes with major public health implications, such as new relapsing patterns, drug resistance and increased virulence. Suitable molecular markers are required for these population genetic studies. Here, we focus on two groups of molecular markers that are commonly used to analyse natural populations of P. vivax. We use markers under selective pressure, for instance, antigen-coding polymorphic genes, and markers that are not under strong natural selection, such as most minisatellite and microsatellite loci. First, we review data obtained using genes encoding for P. vivax antigens: circumsporozoite protein, merozoite surface proteins 1 and 3α, apical membrane antigen 1 and Duffy binding antigen. We next address neutral or nearly neutral molecular markers, especially microsatellite loci, providing a complete list of markers that have already been used in P. vivax populations studies. We also analyse the microsatellite loci identified in the P. vivax genome project. Finally, we discuss some practical uses for P. vivax genotyping, for example, detecting multiple-clone infections and tracking the geographic origin of isolates.

Culex quinquefasciatus vitellogenesis: morphological and biochemical aspects

The vitellogenic process in Culex quinquefasciatus, which is triggered by a blood meal, involves the synthesis, distribution and storage of the nutrients necessary for embryo development. The fat body of an adult female Cx. quinquefasciatus revealed two cell types: large trophocytes and small, eosinophilic, "oenocyte-like" cells, which show no morphological changes throughout the gonotrophic cycle. Trophocytes, which only begin to synthesise vitellogenin (Vg) 12 h post-blood meal (PBM), undergo a series of morphological changes following engorgement. These changes include the expansion of the rough endoplasmic reticulum (RER) and Golgi complex, which are later destroyed by autophagosomes. At 84 h PBM, trophocytes return to their pre-engorgement morphology. The ovarian follicles of non-blood-fed Cx. quinquefasciatus contain a cluster of eight undifferentiated cells surrounded by follicular epithelium. After engorgement, the oocyte membrane facing the perioocytic space increases its absorptive surface by microvilli development; large amounts of Vg and lipids are stored between 24 and 48 h PBM. Along with yolk storage in the oocyte, follicular cells exhibit the development of RER cisternae and electron-dense granules begin to fill the perioocytic space, possibly giving rise to endochorion. Later in the gonotrophic cycle, electron-dense vesicles, which are possible exochorion precursors, fuse at the apical membrane of follicular cells. This fusion is followed by follicular cell degeneration.

Distribuição de tensões em mini-implantes ortodônticos

O objetivo deste trabalho foi avaliar a distribuição de tensões na resina em contato com os filetes de roscas de mini-implantes cilíndricos e cônicos, submetidos à carga lateral e torção de inserção. Um modelo fotoelástico foi confeccionado com gelatina transparente, para simular o osso alveolar. O modelo foi observado com um polariscópio plano e fotografado antes e após a ativação dos mini-implantes com força lateral e de inserção. A aplicação de cargas laterais provocou momentos fletores nos mini-implantes, aparecimento de franjas isocromáticas ao longo dos filetes do corpo dos mini-implantes e no ápice. Quando foi aplicado o torque de inserção, verificou-se a concentração de tensões próxima ao ápice. Concluiu-se que: (1) o mini-implante cilíndrico apresentou maior concentração de tensões no ápice, e (2) o mini-implante cônico apresentou maior concentração de tensões nos filetes de rosca apicais.

Análise de atitudes de professoras do ensino fundamental no que se refere à educação inclusiva

Este artigo, que tem como referência a teoria crítica, relata o resultado de uma pesquisa sobre atitudes em relação à educação inclusiva, realizada com doze professoras do 5º ano do ensino fundamental; dentre elas, seis tinham experiência de ter em sala de aula alunos com deficiência, e seis não tinham. A análise qualitativa dos dados decorrentes do levantamento realizado em seis escolas da rede municipal de Campo Grande (MS) demonstrou que, de modo geral, as professoras foram favoráveis à educação inclusiva. Contudo, ficou evidente a expressão de atitudes preconceituosas veladas ou explícitas no âmbito escolar. A formação para experiência com quem é "diferente" ainda encontra barreiras por conta do preconceito e da discriminação presentes nesta sociedade que tem como lógica uma "inclusão marginal". Em suas manifestações, apontaram dificuldades de trabalhar com alunos com deficiência intelectual severa; assinalaram que a responsabilidade de trabalhar sob a predominância da educação centrada no desempenho, com foco na inserção das pessoas no mercado de trabalho, gerava um "sentimento de impotência". Diante das contradições existentes, a educação inclusiva não deixa de evidenciar a presença das injustiças que ainda se apresentam no processo educativo. Apesar disso, foram indicados vários elementos favoráveis a essa forma de educação, que não é dissociada de movimentos sociais mais amplos.

Novos táxons e novo registro em Hemilophini (Coleoptera: Cerambycidae: Lamiinae)

Novas espécies descritas e ilustradas: Adesmus nigrolineatus sp. nov. do México (Oaxaca). Da Costa Rica: A. moruna sp. nov. (Heredia); Corcovado bezarki sp. nov. (Guanacaste); Alampyris fuscus sp. nov. (Guanacaste), Cariua gen. nov. espécie-tipo C. sulphurea sp. nov., (Guanacaste). Da Bolívia: Phoebemima albomaculata sp. nov. (Cochabamba); Ipepo gen. nov. espécie-tipo I. dilatatus sp. nov. (Santa Cruz). Do Brasil: Adesmus facetus sp. nov. e Canarana arguta sp. nov. (Rondônia). É acrescentado novo registro na Costa Rica para Piruanycha pitilla Galileo & Martins, 2005. As três espécies novas de Adesmus distinguem-se: A. nigrolineatus sp. nov. pelas faixas longitudinais de tegumento preto nos élitros; A. moruna sp. nov. pelos élitros inteiramente pretos; A. facetus pelas faixas oblíquas de pubescência branca após o meio dos élitros além das manchas do quarto apical e das epipleuras. Phoebemima albomaculata sp. nov. caracteriza-se pela mancha basal de pubescência branca dos élitros estendendo-se sobre a sutura. Corcovado bezarki sp. nov. distingue-se pelo escapo preto e antenômeros esbranquiçados. Canarana arguta sp. nov. tem o protórax e os urosternitos I a IV cobertos por densa pubescência amarelada. Alampyris fusca sp. nov. difere de A. cretaria principalmente pelo antenômero III mais longo que o escapo. Cariua sulphurea sp. nov. separa-se pela presença de urosternitos revestidos por pubescência branca compacta e Ipepo dilatatus sp. nov. caracteriza-se pelos élitros tri-carenados.

Culicídeos em bromélias: diversidade de fauna segundo influência antrópica, litoral de São Paulo

OBJETIVO: Comparar a diversidade da fauna de culicídeos em bromélias de solo segundo ambientes urbano, periurbano e mata primitiva. MÉTODOS: O estudo foi realizado no município de Ilhabela, litoral norte do estado de São Paulo, em tanques de bromélias de ambientes urbano, periurbano e mata. Realizaram-se coletas de imaturos quinzenalmente, de março de 1998 a julho de 1999. A presença e freqüência de espécies nos diferentes ambientes foram comparadas com base em estimativas da diversidade para medir a riqueza, dominância e análise de variância (ANOVA). RESULTADOS: Coletaram-se 31.134 formas imaturas de mosquitos nas bromélias, distribuídas em sete gêneros e 37 espécies. O ambiente urbano registrou maior abundância, 14.575 indivíduos, seguido do periurbano com 10.987, e a mata, com o menor número de exemplares, 5.572. Foram coletadas 30 espécies no habitat urbano, 32 no periurbano e 33 na mata. As espécies dominantes foram Culex (Microculex) pleuristriatus nos ambientes urbano e periurbano, e Culex ocellatus na mata. De acordo com teste ANOVA a freqüência de mosquitos em bromélias não foi diferente entre os ambientes pesquisados (F=0,5564; p=0,5769). A diversidade de espécies na mata foi maior, e semelhante entre periurbano e urbano. CONCLUSÕES: A composição específica de culicídeos em bromélias de solo mostrou-se diversificada, sendo maior naquelas de ambiente de mata. As espécies dominantes foram Cx. (Mcx.) pleuristriatus e Cx. ocellatus.

Establishment of a protocol for obtention of neuronal stem cells lineages from the dog olfactory epithelium

A morphological and cell culture study from nasal mucosa of dogs was performed in order to establish a protocol to obtain a cell population committed to neuronal lineage, as a proposal for the treatment of traumatic and degenerative lesions in these animals, so that in the future these results could be applied to the human species. Twelve mongrel dogs of 60-day aged pregnancy were collected from urban pound dogs in São Paulo. Tissue from cribriform ethmoidal lamina of the fetuses was collected at necropsy under sterile conditions around 1h to 2h postmortem by uterine sections and sections from the fetal regions described above. Isolated cells of this tissue were added in DMEM/F-12 medium under standard conditions of incubation (5% CO², >37ºC). Cell culture based on isolated cells from biopsies of the olfactory epithelium showed rapid growth when cultured for 24 hours, showing phase-bright sphere cells found floating around the fragments, attached on culture flasks. After 20 days, a specific type of cells, predominantly ellipsoids or fusiform cells was characterized in vitro. The indirect immunofluorescence examination showed cells expressing markers of neuronal precursors (GFAP, neurofilament, oligodendrocyte, and III â-tubulin). The cell proliferation index showed Ki67 immunostaining with a trend to label cell groups throughout the apical region, while PCNA immunostaining label predominantly cell groups lying above the basal lamina. The transmission electron microscopy from the olfactory epithelium of dogs revealed cells with electron-dense cytoplasm and preserving the same distribution as those of positive cell staining for PCNA. Metabolic activity was confirmed by presence of euchromatin in the greatest part of cells. All these aspects give subsidies to support the hypothesis about resident progenitor cells among the basal cells of the olfactory epithelium, committed to renewal of these cell populations, especially neurons.

Embryonic, Larval, and Juvenile Development of the Sea Biscuit Clypeaster subdepressus (Echinodermata: Clypeasteroida)

Sea biscuits and sand dollars diverged from other irregular echinoids approximately 55 million years ago and rapidly dispersed to oceans worldwide. A series of morphological changes were associated with the occupation of sand beds such as flattening of the body, shortening of primary spines, multiplication of podia, and retention of the lantern of Aristotle into adulthood. To investigate the developmental basis of such morphological changes we documented the ontogeny of Clypeaster subdepressus. We obtained gametes from adult specimens by KCl injection and raised the embryos at 26 degrees C. Ciliated blastulae hatched 7.5 h after sperm entry. During gastrulation the archenteron elongated continuously while ectodermal red-pigmented cells migrated synchronously to the apical plate. Pluteus larvae began to feed in 3 d and were similar to 20 d old at metamorphosis; starved larvae died 17 d after fertilization. Postlarval juveniles had neither mouth nor anus nor plates on the aboral side, except for the remnants of larval spicules, but their bilateral symmetry became evident after the resorption of larval tissues. Ossicles of the lantern were present and organized in 5 groups. Each group had 1 tooth, 2 demipyramids, and 2 epiphyses with a rotula in between. Early appendages consisted of 15 spines, 15 podia (2 types), and 5 sphaeridia. Podial types were distributed in accordance to Loven's rule and the first podium of each ambulacrum was not encircled by the skeleton. Seven days after metamorphosis juveniles began to feed by rasping sand grains with the lantern. Juveniles survived in laboratory cultures for similar to 9 months and died with <500 mu m wide, a single open sphaeridium per ambulacrum, aboral anus, and no differentiated food grooves or petaloids. Tracking the morphogenesis of early juveniles is a necessary step to elucidate the developmental mechanisms of echinoid growth and important groundwork to clarify homologies between irregular urchins.

The Relationship between Population Structure and Aluminum Tolerance in Cultivated Sorghum

Background: Acid soils comprise up to 50% of the world's arable lands and in these areas aluminum (Al) toxicity impairs root growth, strongly limiting crop yield. Food security is thereby compromised in many developing countries located in tropical and subtropical regions worldwide. In sorghum, SbMATE, an Al-activated citrate transporter, underlies the Alt(SB) locus on chromosome 3 and confers Al tolerance via Al-activated root citrate release. Methodology: Population structure was studied in 254 sorghum accessions representative of the diversity present in cultivated sorghums. Al tolerance was assessed as the degree of root growth inhibition in nutrient solution containing Al. A genetic analysis based on markers flanking Alt(SB) and SbMATE expression was undertaken to assess a possible role for Alt(SB) in Al tolerant accessions. In addition, the mode of gene action was estimated concerning the Al tolerance trait. Comparisons between models that include population structure were applied to assess the importance of each subpopulation to Al tolerance. Conclusion/Significance: Six subpopulations were revealed featuring specific racial and geographic origins. Al tolerance was found to be rather rare and present primarily in guinea and to lesser extent in caudatum subpopulations. Alt(SB) was found to play a role in Al tolerance in most of the Al tolerant accessions. A striking variation was observed in the mode of gene action for the Al tolerance trait, which ranged from almost complete recessivity to near complete dominance, with a higher frequency of partially recessive sources of Al tolerance. A possible interpretation of our results concerning the origin and evolution of Al tolerance in cultivated sorghum is discussed. This study demonstrates the importance of deeply exploring the crop diversity reservoir both for a comprehensive view of the dynamics underlying the distribution and function of Al tolerance genes and to design efficient molecular breeding strategies aimed at enhancing Al tolerance.

Plasmodium vivax circumsporozoite genotypes: a limited variation or new subspecies with major biological consequences?

Background: Plasmodium vivax circumsporozoite variants have been identified in several geographical areas. The real implication of the genetic variation in this region of the P. vivax genome has been questioned for a long time. Although previous studies have observed significant association between VK210 and the Duffy blood group, we present here that evidences of this variation are limited to the CSP central portion. Methods: The phylogenetic analyses were accomplished starting from the amplification of conserved domains of 18 SSU RNAr and Cyt B. The antibodies responses against the CSP peptides, MSP-1, AMA-1 and DBP were detected by ELISA, in plasma samples of individuals infected with two P. vivax CS genotypes: VK210 and P. vivax-like. Results: These analyses of the two markers demonstrate high similarity among the P. vivax CS genotypes and surprisingly showed diversity equal to zero between VK210 and P. vivax-like, positioning these CS genotypes in the same clade. A high frequency IgG antibody against the N- and C-terminal regions of the P. vivax CSP was found as compared to the immune response to the R- and V-repetitive regions (p = 0.0005, Fisher's Exact test). This difference was more pronounced when the P. vivax-like variant was present in the infection (p = 0.003, Fisher's Exact test). A high frequency of antibody response against MSP-1 and AMA-1 peptides was observed for all P. vivax CS genotypes in comparison to the same frequency for DBP. Conclusions: This results target that the differences among the P. vivax CS variants are restrict to the central repeated region of the protein, mostly nucleotide variation with important serological consequences.

Effect of Laser Phototherapy on the Release of TNF-alpha and MMP-1 by Endodontic Sealer-Stimulated Macrophages

Objective: Our aim was to analyze the effect of laser phototherapy on the secretory activity of macrophages activated by interferon-gamma (IFN-gamma) and lipopolysaccharide (LPS), and stimulated by substances leached from an epoxy resin-based sealer (AH-Plus) and a calcium hydroxide-based sealer (Sealapex). Background Data: Laser phototherapy can modulate the inflammatory process, improving wound healing. This type of therapy could be useful for modulating postoperative symptoms seen after endodontic treatment. Materials and Methods: Cytotoxicity was indirectly assessed by measuring mitochondrial activity. Macrophages were stimulated by the leached substances or not (controls), and the groups were then irradiated or not. The secretion of pro-inflammatory cytokines (TNF-alpha and MMP-1) was analyzed using ELISA. Two irradiations at 6-h intervals were done with an As-Ga-Al diode laser (780 nm, 70 mW, spot size 4.0 mm(2), 3 J/cm(2), for 1.5 sec) in contact mode. Results: The sealers were non-cytotoxic to macrophages. The production of TNF-alpha was significantly decreased by laser phototherapy, regardless of experimental group. The level of secretion of MMP-1 was similar in all groups. Conclusion: Based on the conditions of this study we concluded that in activated macrophages, laser phototherapy impairs the secretion of the pro-inflammatory cytokine TNF-alpha, but has no influence on MMP-1 secretion.