Morfoanatomia da flor de cinco espécies de Galipea Aubl. e seu significado na evolução de flores tubulosas entre as Rutaceae neotropicais

A maioria dos gêneros da subtribo neotropical Galipeinae (tribo Galipeeae, Rutoideae) tem flores tubulosas, com várias formas e graus de conação e adnação. Galipea e outros gêneros na subtribo apresentam apenas duas anteras férteis mais cinco ou mais estaminódios, o que é intrigante porque na tribo predominam flores pentâmeras isostêmones. Visando elucidar a condição anatômica dessas características e estabelecer estados acurados para caracteres em análises filogenéticas, um estudo morfoanatômico de flores de cinco espécies de Galipea foi realizado, buscando os padrões de vascularização, posição, e união dos segmentos da flor. Destacam-se os resultados: 1) um tubo floral genuíno se forma no terço basal da flor por conação dos filetes e adnação desse tubo estaminal às pétalas; 2) as pétalas são distalmente coerentes umas às outras e aderentes aos filetes por meio de entrelaçamento de tricomas densos - um caso de pseudossimpetalia; 3) dentre as cinco (às vezes seis) estruturas tratadas como estaminódios, apenas as três externas são de fato homólogas a estames esterilizados, as demais surgem como ramificações adaxiais das pétalas; 4) os carpelos são peltados, congenitalmente conatos axial e lateralmente da base do ovário até o nível das placentas, e no estilete e estigma; na zona mediana e superior do ovário eles são unidos apenas posgenitalmente, com a epiderme diferenciada de carpelos contíguos e suturas evidentes na região ventral de cada carpelo; 5) a vascularização do disco sugere origem receptacular. As implicações desses dados para o entendimento da evolução das flores tubulosas em Galipea e grupos relacionados são discutidas.

Two-dimensional gel electrophoretic protein profile analysis during seed development of Ocotea catharinensis: a recalcitrant seed species

The aim of the present work was to characterize changes in the protein profile throughout seed development in O. catharinensis, a recalcitrant species, by two-dimensional gel electrophoresis. Protein extraction was undertaken by using a thiourea/urea buffer, followed by a precipitation step with 10% TCA. Comparative analysis during seed development showed that a large number of proteins were exclusively detected in each developmental stage. The cotyledonary stage, which represents the transition phase between embryogenesis and the beginning of metabolism related to maturation, presents the highest number of stage-specific spots. Protein identification, through MS/MS analysis, resulted in the identification of proteins mainly related to oxidative metabolism and storage synthesis. These findings contribute to a better understanding of protein metabolism during seed development in recalcitrant seeds, besides providing information on established markers that could be useful in defining and improving somatic embryogenesis protocols, besides monitoring the development of somatic embryos in this species.

Correlation of meta 1 expression with culture stage, cell morphology and infectivity in Leishmania (Leishmania) amazonensis promastigotes

The parasitic protozoan Leishmania (Leishmania) amazonensis alternates between mammalian and insect hosts. In the insect host, the parasites proliferate as procyclic promastigotes andthen differentiate into metacyclic infective forms. The meta 1 gene is preferentially expressed during metacyclogenesis. Meta 1 expression profile determination along parasite growth curves revealed that the meta 1 mRNA level peaked at the early stationary phase then decreased to an intermediate level. No correlation was observed between meta 1 expression and infectivity. Conversely, infectivity correlated with the increase of apoptotic cells in the late stationary phase.

Reproductive behavior, development and eye regression in the cave armored catfish, Ancistrus cryptophthalmus Reis, 1987 (Siluriformes: Loricariidae), breed in laboratory

The troglobitic armored catfish, Ancistrus cryptophthalmus (Loricariidae, Ancistrinae) is known from four caves in the São Domingos karst area, upper rio Tocantins basin, Central Brazil. These populations differ in general body shape and degree of reduction of eyes and of pigmentation. The small Passa Três population (around 1,000 individuals) presents the most reduced eyes, which are not externally visible in adults. A small group of Passa Três catfish, one male and three females, reproduced spontaneously thrice in laboratory, at the end of summertime in 2000, 2003 and 2004. Herein we describe the reproductive behavior during the 2003 event, as well as the early development of the 2003 and 2004 offsprings, with focus on body growth and ontogenetic regression of eyes. The parental care by the male, which includes defense of the rock shelter where the egg clutch is laid, cleaning and oxygenation of eggs, is typical of many loricariids. On the other hand, the slow development, including delayed eye degeneration, low body growth rates and high estimated longevity (15 years or more) are characteristic of precocial, or K-selected, life cycles. In the absence of comparable data for close epigean relatives (Ancistrus spp.), it is not possible to establish whether these features are an autapomorphic specialization of the troglobitic A. cryptophthalmus or a plesiomorphic trait already present in the epigean ancestor, possibly favoring the adoption of the life in the food-poor cave environment. We briefly discuss the current hypotheses on eye regression in troglobitic vertebrates.

Immature stages of the moth Lobeza dentilinea (Lepidoptera: Notodontidae), with redescription of the species based on male and female morphology

Egg and pupa of Lobeza dentilinea Schaus, 1901 are described and illustrated for the first time. Eggs are smooth, dome-shaped, and greenish at oviposition. Last instar larvae have an aposematic coloration and the chaetotaxy is very similar to other notodontines, except for the number of lateral setae: L. dentilinea has three instead of four lateral setae on abdominal segments A3-A6. Pupae are light brown and typical of the family, with the last abdominal segments broadly round. Evidence from the adult morphology supporting the placement of the genus in Notodontinae includes proboscis smaller than the length of the head, epiphysis with more than half the length of tibia, tarsal claws simple, and labial palpi short. Male and female are confidently associated, and a redescription of the species is presented based on both sexes. Larvae of L. dentilinea are here recorded feeding on a Melastomataceae.

O enigma da "reação espermatofórica": breve síntese do conhecimento sobre a estrutura e o funcionamento dos espermatóforos dos cefalópodes (Mollusca: Cephalopoda)

Cefalópodes coleóides (lulas, sépias e polvos) produzem espermatóforos muito complexos que são transferidos à fêmea durante a cópula por meio do hectocótilo, um apêndice modificado nos machos. Durante a transferência à fêmea, ocorre a chamada "reação espermatofórica", complexo processo de evaginação do aparato ejaculatório do espermatóforo, que conduz à exteriorização da massa espermática e corpo cimentante. A presente revisão sintetiza o conhecimento acerca da morfologia e funcionamento desta estrutura exclusiva dos coleóides, identificando lacunas e definindo estratégias que possibilitem avanços na área. Poucos trabalhos abordam com detalhes a morfologia e anatomia funcional dos espermatóforos dos cefalópodes, grande parte do conhecimento acerca da estrutura do espermatóforo tendo sido gerada por trabalhos clássicos do século XIX e início do século XX. Investigações acerca do funcionamento dos espermatóforos são consideravelmente mais raras, estando o conhecimento básico sobre a reação espermatofórica restrito a apenas 19 espécies de coleóides. A revisão da literatura especializada permite sugerir que existem dois tipos básicos de fixação de espermatóforos em Decapodiformes (lulas e sepióides): fixação superficial e implante profundo (ou intra-dérmico). Na fixação superficial, comum em diversas espécies (e.g., Loliginidae, Sepiidae, Ommastrephidae), a base dos espermatângios é aderida ao tecido-alvo aparentemente por meio do corpo cimentante, a partir de substâncias adesivas e, em alguns casos, estruturas de fixação. No implante profundo, comum em alguns grupos de lulas oceânicas e de águas profundas (e.g., Architeuthidae, Cranchiidae, Octopoteuthidae, Sepiolidae), os espermatóforos implantam-se inteiramente no corpo da fêmea, de forma autônoma. Permanece desconhecido o mecanismo responsável pelo implante profundo. Em Octopodiformes (polvos), o espermatóforo é inserido no gonoduto feminino, alcançando a glândula oviducal, onde estão localizadas as espermatecas, ou a cavidade do ovário. Como o funcionamento extracorpóreo dos espermatóforos depende exclusivamente da intrincada estrutura e organização de seus componentes (e.g., membranas e túnicas), somente investigações detalhadas dessas estruturas proverão as bases para a compreensão do funcionamento e da exata função do complexo espermatóforo dos coleóides. Recomenda-se o desenvolvimento de um protocolo simples e eficiente para coloração e preparação total de espermatóforos, de forma que seja possível expandir as descrições morfológicas do espermatóforo em estudos taxonômicos e anatômicos, permitindo, portanto, ampliação do conhecimento acerca desta enigmática estrutura.

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.

Morphology and reproduction of Mesophyllum erubescens (Foslie) Me. Lemoine (Corallinales, Rhodophyta) from Southern Brazil

The genus Mesophyllum Me. Lemoine includes around 147 species, of which only three have been referred to the Brazilian coast. Mesophyllum erubescens was originally described from Fernando de Noronha Archipelago, Brazil (type locality). Here we present the first detailed description of M. erubescens based on Brazilian material. Samplings were made through scuba diving at the Biological Marine Reserve of Arvoredo Island, Santa Catarina. The relations of M. erubescens with other similar species, especially from the American Atlantic studied by W.R. Taylor are discussed.

Cation size effects-modified phase and PTCR development in Er3+ and Ca2+ co-doped BaTiO3 ceramics during sintering

Development of the positive temperature coefficient of resistivity (PTCR) in Er3+ and Ca2+ co-doped ferroelectric BaTiO3 was studied in this work, with Er3+ being used to act as a donor doping. Irrespective of all the materials showing high densities after sintering at 1200 to 1300 ºC, these revealed insulator at the lowest sintering temperature, changing to semiconducting and PTCR-type materials only when the sintering temperature was further increased. Observations from X-ray diffraction help correlating this effect with phase development in this formulated (Ba,Ca,Er)TiO3 system, considering the formation of initially two separated major (Ba,Ca)TiO3- and minor (Ca,Er)TiO3-based compounds, as a consequence of cation size-induced stress energy effects. Thus, appearance and enhancement here of the semiconducting and PTCR responses towards higher sintering temperatures particularly involve the incorporation of Er3+ into the major phase, rendering finally possible the generation and "percolative-like" migration of electrons throughout the whole material.

Aerobic and anaerobic metabolism for the zebrafish, Danio rerio, reared under normoxic and hypoxic conditions and exposed to acute hypoxia during development

In order to verify the influence of chronic and acute ambient oxygen levels from egg to adult stage of the zebrafish, in vivo oxygen consumption (MO2), critical tensions of oxygen (Pcrit), heart rate (fH) and total body lactate concentration (Lc) were determined for Danio rerio (Hamilton, 1822) raised at 28 °C under normoxic (7.5 mgO2.L-1 or 80 mm.Hg-1) and hypoxic conditions (4.3 mgO2.L-1) and exposed to acute hypoxia during different developmental stages. Our findings confirmed that very early stages do not respond effectively to ambient acute hypoxia. However, after the stage corresponding to the age of 30 days, D. rerio was able to respond to acute hypoxia through effective physiological mechanisms involving aerobic and anaerobic metabolism. Such responses were more efficient for the fishes reared under hypoxia which showed that D. rerio survival capability increased during acclimation to mild hypoxia. Measurements of body mass and length showed that moderate hypoxia did not affect growth significantly until the fish reached the stage of 60 days. Moreover, a growth delay was verified for the hypoxic-reared animals. Also, the D. rerio eggs-to-larvae survival varied from 87.7 to 62.4% in animals reared under normoxia and mild hypoxia, respectively. However, the surviving animals raised under moderated hypoxia showed a better aptitude to regulate aerobic and anaerobic capacities when exposed to acute hypoxia.

Development of instrumentation for amperometric and coulometric detection using ultramicroelectrodes

In this work it is presented the development of a simple, portable and inexpensive instrumentation for amperometric and coulometric detection in different analytical instrumentation systems utilizing ultramicroelectrodes. The software, developed in LabVIEW 7.1TM, is capable to carry out three main detection techniques (amperometric, pulsed amperometric and coulometric detection) and a voltammetric technique (cyclic voltammetry). The instrumentation was successfully evaluated using the following systems: cyclic voltammograms of metallic electrodes in alkaline solutions, flow electrochemical detection of glucose and glycine and direct determination of herbicide glyphosate (electrochemical detection coupled to HPLC).

Development and validation of a simple and rapid capillary zone electrophoresis method for determination of nnrti nevirapine in pharmaceutical formulations

A simple and fast capillary zone electrophoresis (CZE) method has been developed and validated for quantification of a non-nucleoside reverse transcriptase inhibitor (NNRTI) nevirapine, in pharmaceuticals. The analysis was optimized using 10 mmol L-1 sodium phosphate buffer pH 2.5, +25 kV applied voltage, hydrodynamic injection 0.5 psi for 5 s and direct UV detection at 200 µm. Diazepam (50.0 µg mL-1) was used as internal standard. Under these conditions, nevirapine was analyzed in approximately less than 2.5 min. The analytical curve presented a coefficient of correlation of 0.9994. Limits of detection and quantification were 1.4 µg mL-1 and 4.3 µg mL-1, respectively. Intra- and inter-day precision expressed as relative standard deviations were 1.4% and 1.3%, respectively and the mean recovery was 100.81%. The active pharmaceutical ingredient was subjected to hydrolysis (acid, basic and neutral) and oxidative stress conditions. No interference of degradation products and tablet excipients were observed. This method showed to be rapid, simple, precise, accurate and economical for determination of nevirapine in pharmaceuticals and it is suitable for routine quality control analysis since CE offers benefits in terms of quicker method development and significantly reduced operating costs.

Development of a flow through photo-reactor to study degradation of organic compounds by Sequential Injection Analysis (SIA)

This work describes a photo-reactor to perform in line degradation of organic compounds by photo-Fenton reaction using Sequential Injection Analysis (SIA) system. A copper phthalocyanine-3,4',4²,4²¢-tetrasulfonic acid tetrasodium salt dye solution was used as a model compound for the phthalocyanine family, whose pigments have a large use in automotive coatings industry. Based on preliminary tests, 97% of color removal was obtained from a solution containing 20 µmol L-1 of this dye.

Epilithic community development on artificial reefs deployed along a cross-shelf environmental gradient off Paraná state, southern Brazil

Concrete modules were deployed on the bottom of the 11, 18 and 30 meters isobaths along a cross-shelf hydrographic gradient off Paraná State, Southern Brazil, with the purpose of studying the colonization of sessile epilithic macroinvertebrates on artificial surfaces. After one year of submersion a total of 63 species of epilithic organisms were identified, dominated by Ostrea puelchana, Chthamalus bisinuatus, Balanus cf spongicola, Astrangia cf rathbuni, Didemnum spp, poryphers and bryozoans. Diversity index and percent cover at reef stations placed at 11, 18 and 30 meters isobaths were respectively 2.28 and 66.7%, 2.79 and 96.6% and 1.66 and 77.4%. Differences of general community structure among the three assemblages were not clearly related to the general environmental conditions at the bottom layers near the reef stations. Turbidity and larval abundance are discussed as important factors affecting colonization processes. Results indicate that depths between 15-20 meters are more suitable for the implementation of large scale artificial reef systems in the inner shelf off Paraná and, possibly, throughout the inner shelves off southern Brazil with similar hydrographic conditions.

Comparative morphology among representatives of main taxa of Scaphopoda and basal protobranch Bivalvia (Mollusca)

This study deals with detailed morphology and anatomy of 4 species of Scaphopoda and 5 species of protobranch Bivalvia. Both classes are traditionally grouped in the taxon Diasoma, which has been questioned by different methodologies, such as molecular and developmental. This study is developed under a phylogenetic methodology with the main concern in performing it in an intelligible and testable methodology. The analyzed Scaphopoda species came from the Brazilian coast and belong to the family Dentaliidae [(1) Coccodentalium carduus; (2) Paradentalium disparile] and Gadiliidae; [(3) Polyschides noronhensis, n. sp. from Fernando de Noronha Archipelago; (4) Gadila braziliensis]. These species represent the main branches of the class Scaphopoda. From protobranch bivalves, representatives of the families Solemyidae [(5) Solemya occidentalis, from Florida; S. notialis, n. sp. from S.E. Brazil], Nuculanidae [(6) Propeleda carpentieri from Florida], and Nuculidae [(7) Ennucula puelcha, from south Brazil] are included. These species represent the main branches of the basal Bivalvia. The descriptions on the anatomy of S. occidentalis and of P. carpentieri are published elsewhere. The remaining are included here, for which a complete taxonomical treatment is performed. Beyond these species, representatives of other taxa are operationally included as part of the ingroup (indices are then shared with them), as a procedure to test the morphological monophyly of Diasoma. These taxa are: two lamellibranch bivalves [(8) Barbatia - Arcidae; (9) Serratina - Tellinidae; both published elsewhere;, and Propilidium (10) Patellogastropoda, and (11) Nautilus, basal Cephalopoda, based on basal taxa. The effective outgroups are (12) Neopilina (Monoplacophora) and (13) Hanleya (Polyplacophora). The phylogenetic analysis based on morphology revealed that the taxon Diasoma is supported by 14 synapomorphies, and is separated from Cyrtosoma (Gastropoda + Cephalopoda). Although they are not the main goal of this paper, the taxa Scaphopoda and Bivalvia are supported by 8 and by 7 synapomorphies respectively. The taxon Protobranchia resulted paraphyletic. Both scaphopod orders resulted monophyletic. The obtained cladogram is: ((((Coccodentalium carduus - Paradentalium disparile) (Polyschides noronhensis - Gadila brasiliensis)) ((Solemya occidentalis - S. notialis) (Propeleda carpenteri (Ennucula puelcha (Barbatia cancellaria - Serratina capsoides))))) (Propilidium curumim - Nautilus pompilius - Lolliguncula brevis)).