Programmed cell death in the larval salivary glands of Apis mellifera (Hymenoptera, Apidae)

The morphological and histochemical features of degeneration in honeybee (Apis mellifera) salivary glands were investigated in 5th instar larvae and in the pre-pupal period. The distribution and activity patterns of acid phosphatase enzyme were also analysed. As a routine, the larval salivary glands were fixed and processed for light microscopy and transmission electron microscopy. Tissue sections were subsequently stained with haematoxylin-eosin, bromophenol blue, silver, or a variant of the critical electrolyte concentration (CEC) method. Ultrathin sections were contrasted with uranyl acetate and lead citrate. Glands were processed for the histochemical and cytochemical localization of acid phosphatase, as well as biochemical assay to detect its activity pattern. Acid phosphatase activity was histochemically detected in all the salivary glands analysed. The cytochemical results showed acid phosphatase in vesicles, Golgi apparatus and lysosomes during the secretory phase and, additionally, in autophagic structures and luminal secretion during the degenerative phase. These findings were in agreement with the biochemical assay. At the end of the 5th instar, the glandular cells had a vacuolated cytoplasm and pyknotic nuclei, and epithelial cells were shed into the glandular lumen. The transition phase from the 5th instar to the pre-pupal period was characterized by intense vacuolation of the basal cytoplasm and release of parts of the cytoplasm into the lumen by apical blebbing; these blebs contained cytoplasmic RNA, rough endoplasmic reticule and, occasionally, nuclear material. In the pre-pupal phase, the glandular epithelium showed progressive degeneration so that at the end of this phase only nuclei and remnants of the cytoplasm were observed. The nuclei were pyknotic, with peripheral chromatin and blebs. The gland remained in the haemolymph and was recycled during metamorphosis. The programmed cell death in this gland represented a morphological form intermediate between apoptosis and autophagy.

Nuclear alterations associated to programmed cell death in larval salivary glands of Apis mellifera (Hymenoptera : Apidae)

The silk glands of bees are a good model for the study of cell death in insects. With the objective to detect the nuclear features during glandular regression stage, larvae at the last instar and pre-pupae were collected and their silk glands were dissected and processed for ultrastructural analysis and histologically for cytochemical and imunocytochemical analysis. The results showed that the cellular nuclei exhibited characteristics of death by atypical apoptosis as well as autophagic cell death. Among the apoptosis characteristic were: nuclear strangulation with bleb formation in some nuclei, DNA fragmentation in most of the nuclei and nucleolar fragmentation. Centripetal chromatin compaction was observed in many nuclei, forming a perichromatin halo differing from typical apoptotic nuclei. With regards to the characteristics of autophagic-programmed cell death, most relevant was the delay in the collapse of many nuclei. (C) 2006 Elsevier Ltd. All rights reserved.

Post-Embryonic Development of Larvae of the Weaver Ant Camponotus textor (Hymenoptera: Formicidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Handicrafts from Jalapao (TO), Brazil, and their relationship to plant anatomy

EICHEMBERG, M. T. and V. L. SCATENA (Universidade Estadual Paulista, Departamento de Botanica, Rio Claro, São Paulo, Brazil). J. Torrey Bot. Soc. 138:34-40. 2011.-Handicrafts from Jalapao (TO), Brazil, and their Relationship to Plant Anatomy. In the state of Tocantins, midwestern Brazil, communities from the region of Jalapao use scapes of "capim dourado" (golden grass - Syngonanthus nitens- Eriocaulaceae) and leaves of "buriti" (Mauritia flexuosa - Arecaceae) to make handicrafts (baskets and ornaments). The predominant biome of this area is cerrado (savanna), with a notable presence of buriti in the "veredas" (swampy forest-like vegetation), and of golden grass, which is one of the most common plants in humid grasslands. These traditional handicrafts represent a significant source of income for local communities. The whole scapes of Syngonanthus nitens are used due to their golden color, which is a reflection of such internal structures as thick walled cells and lignin in the epidermis and cortex. The strips called "seda" (silk) used to sew the scapes in the making of handicrafts come from young leaves of Mauritia,flexuosa. They are constituted by the adaxial epidermis and bundles of subepidermic fibers, both showing thick-walled cells. Since the cells of the bundles of sclerenchymatic fibers from the abaxial surface of buriti leaves present stegmata containing silica bodies, their mechanical properties are less adapted to the production of "silk", justifying the use of the leaf adaxial surface. Anatomical characteristics such as the thickening and composition of the cell walls of both species together with sociocultural factors, allow a better knowledge of the use of plant structures in the making of handicrafts.

The chelation of metal ions by the acylpolyamine toxins from the web-spider Nephilengys cruentata: effects in the intoxication/detoxification of preys

Orb-web-spiders present a series of different strategies for prey capture, involving the use of different types of silk for web building, the use of adhesive traps in the webs, the secretion of toxic compounds to the spider's preys in the adhesive coating of the capture web and the biosynthesis of a wide range of structurally related acylpolyamine toxins in their venoms. The polyamine toxins usually block neuromuscular junctions and/or the central nervous system (CNS) of Arthropods, targeting specially the ionotropic glutamate receptors; this way these toxins are used are as chemical weapons to kill / paralyze the spider's prey. Polyamine toxins contain many azamethylene groups involved with the chelation of metal ions, which in turn can interact with the glutamate receptors, affecting the toxicity of these toxins. It was demonstrated that the chelation of Ni+2, Fe+2, Pb+2, Ca+2 and Mg+2 ions by the desalted crude venom of Nephilengys cruentata and by the synthetic toxin JSTX-3, did not cause any significant change in the toxicity of the acylpolyamine toxins to the model-prey insect (honeybees). However, it was also reported that the chelation of Zn+2 ions by the acylpolyamines potentiated the lethal / paralytic action of these toxins to the honeybees, while the chelation of Cu+2 ions caused the inverse effect. Atomic absorption spectrometry and Plasma-ICP analysis both of N.cruentata venom and honeybee's hemolymph revealed that the spider's venom concentrates Zn+2 ions, while the honeybee's hemolymph concentrates Cu+2 ions. These results are suggesting that the natural accumulation of Zn+2 ions in N. cruentata venom favors the prey catching and/or its maintenance in the web, while the natural accumulation of Cu+2 ions in prey's hemolymph minimizes the efficiency of the acylpolyamine toxins as killing/paralyzing tool.

Description of the Immature and Adult Stages of Ectatomma vizottoi (Formicidae: Ectatomminae)

We estimated the number of larval instars of the ant Ectatomma vizottoi (Ectatomminae), by measuring the maximum width of the head capsules of 208 larvae and the morphology of the immature stages (eggs, larvae and pupae) and adults. There are three larval instars during the post-embryonic development. The reproductive eggs are dark brown. Hairs are present beginning with the first instar, are uniformly distributed over the larval body, and do not vary in length in the three instars. Pupae are protected by a light-brown silk cocoon. Adults of the worker and queen castes can be differentiated by size.

Reproductive behavior and seasonal occurrence of Psecas viridipurpureus (Salticidae, Araneae)

O comportamento reprodutivo e a ocorrência sazonal de Psecas viridipurpureus foram estudados na Estação Ecológica do Noroeste Paulista, uma pequena área de conservação na região noroeste do Estado de São Paulo, Brasil (49º22'50W e 20º48'36S). P. viridipurpureus ocorreu em gravatás, uma bromeliácea (Bromelia balansae, Bromeliaceae) que não acumula água de chuva. Durante a exibição de corte, o casal ocupou a região mediana das folhas de gravatás, com o macho sempre localizado em uma posição superior à da fêmea. Os machos de P. viridipurpureus apresentaram um comportamento de corte complexo, que incluiu cinco padrões motores. O comportamento de corte e a cópula ocorreram preponderantemente durante a estação chuvosa e o recrutamento de jovens entre dezembro e julho. O abrigo de P. viridipurpureus difere do padrão de Salticidae, pois as ootecas são recobertas por uma cobertura de seda plana e não são incluídas em casulos.

Maternal care in a neotropical jumping spider (Salticidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Porous titanium for biomedical applications: An experimental study on rabbits

Objective: The aim of this study was to carry out an in vivo assessment of bone ingrowth in two different types of porous titanium -the first being completely porous, and the second with a porous surface and dense nucleus, manufactured by powder metallurgy- and to evaluate their mechanical properties. Study design: Ten scaffolds from each group were submitted to metallographic analysis and compression tests. Next, two scaffolds of each type were inserted into 14 rabbits, which were sacrificed 8 weeks after surgery. The samples were submitted for histological examination. Results: Metallographic analysis revealed interconnected pores, and the average interconnected pore diameter was about 360 mm, with 36% total porosity. The totally porous titanium samples and the titanium samples with porous surface and dense nucleus showed an average compressive strength of 16.19 MPa and 69.27 MPa, respectively. After 8 weeks, the animals showed bone ingrowth, even into the most internal pores. Conclusions: The pore morphology was effective in permitting bone ingrowth in both groups. Titanium scaffolds with a porous surface and dense nucleus showed the best mechanical properties and most adequate interface.

Relação entre características morfológicas da cariopse e fusariose em milho

Fusarium moniliforme causa sérios prejuízos no rendimento do milho. Suspeita-se que a entrada do fungo na cariopse ocorra pela cicatriz do estilete, e que este fato e o nível de infestação por F. moniliforme estejam relacionados com características morfológicas da cariopse. O objetivo deste trabalho foi identificar características morfológicas que conferem resistência a F. moniliforme em cariopses de milho. Cariopses de seis linhagens de milho, com e sem inoculação do patógeno, foram avaliadas utilizando-se de estereomicroscópio e o microscópio eletrônico de varredura. Características morfológicas da cariopse, tais como: tegumento com saliências e reentrâncias acentuadas, pericarpo pouco espesso, amido menos compacto e presença do canal estilar favorecem a penetração de F. moniliforme.

Enriquecimento da dieta do bicho-da-seda com extrato hidrossolúvel de soja

O objetivo deste trabalho foi avaliar se a suplementação da dieta do bicho-da-seda com extrato hidrossolúvel de soja (EHS) interfere na produção de casulos e no consumo de folhas de amoreira (Morus alba L.) de diferentes cultivares pelas lagartas. O delineamento experimental utilizado foi o inteiramente casualizado em esquema fatorial 4x5, com quatro cultivares de amoreira ('IZ64', 'IZ56/4', 'FM86' e 'FMSM') e cinco dietas (folhas não tratadas, folhas umedecidas com água, folhas enriquecidas com EHS, em três níveis: suplementação da dieta com 5, 10 e 20% de PB). O EHS foi aspergido sobre as folhas, tendo a água como agente dispersante. O consumo foi verificado pela diferença entre a quantidade de folhas fornecidas e a quantidade não consumida pelas lagartas. A ingestão de alimento apresentou-se de forma quadrática, diminuindo à medida que o EHS foi adicionado à dieta. Houve interação dos fatores cultivar e dieta. O fornecimento de folhas de amoreira com acréscimo proteico de 10% por meio do EHS melhorou a qualidade dos casulos do bicho-da-seda com a utilização das cultivares 'IZ56/4' e 'FMSM'. A pulverização de EHS nas folhas da cultivar 'IZ64' prejudicou a produção de casulos. Lagartas do bicho-da-seda, alimentadas com folhas de amoreira sem suplementação, produziram cascas séricas mais pesadas com folhas das cultivares 'IZ64' e 'FMSM'. A aspersão de água nas folhas incrementou o peso de cascas séricas produzidas a partir da cultivar 'FM86'. A suplementação da dieta com EHS diminui o consumo da lagarta do bicho-da-seda. A aplicação de EHS influencia a produção de casulos em função da cultivar utilizada.

Híbridos de amoreira adubados com matéria orgânica e gesso agrícola na produção de bicho-da-seda

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Polímeros biorreabsorvíveis como substrato para cultura de células e engenharia tecidual

Biomateriais poliméricos são desenvolvidos para uso como substitutos de tecidos danificados e/ou estimular sua regeneração. Uma classe de biomateriais poliméricos são os biorreabsorvíveis, compostos que se decompõem tanto in vitro quanto in vivo. São empregados em tecidos que necessitam de um suporte temporário para sua recomposição tecidual. Dentre os vários polímeros biorreabsorvíveis, destacam-se os alfa-hidróxi ácidos, entre eles, diferentes composições do poli(ácido lático) (PLA), como o poli(L-ácido lático) (PLLA), poli(D-ácido lático) (PDLA), poli(DL-ácido lático) (PDLLA), além do poli(ácido glicólico) (PGA) e da policaprolactona (PCL). Estes polímeros são considerados biorreabsorvíveis por apresentarem boa biocompatibilidade e os produtos de sua decomposição serem eliminados do corpo por vias metabólicas. Diversas linhas de pesquisa mostram que os diferentes substratos à base de PLA estudados não apresentam toxicidade, uma vez que as células são capazes de crescer e proliferar sobre eles. Além disso, diversos tipos de células cultivadas sobre diferentes formas de PLA são capazes de se diferenciarem sobre os diferentes polímeros e passar a produzir componentes de matriz extracelular. Neste trabalho, é revisada a utilização de substratos à base de alfa-hidróxi ácidos, com destaque para diferentes formas de PLA, utilizados como substratos para cultura de células, bem como suas aplicações.

Respostas da Tilápia do Nilo (Oreochromis niloticus) à atratividade e palatabilidade de ingredientes utilizados na alimentação de peixes

Avaliaram-se as respostas da tilápia do Nilo à atratividade e palatabilidade de 14 ingredientes. O método utilizado foi de dupla escolha, comparando-se cada ingrediente peletizado à uma ração controle. Foram empregados quatro aquários (750 litros) contendo, cada um, três alevinos e dois comedouros instalados nos cantos direito e esquerdo, sendo registrados tempo decorrido entre colocação do ingrediente e chegada dos animais aos comedouros, freqüência de visitas aos comedouros, número de grânulos ingeridos e freqüência de ejeções. Os tratamentos foram comparados pela prova não-paramétrica de Kruskal-Wallis e a comparação dos ingredientes para o conjunto de variáveis, por intermédio da Análise de Agrupamento e Análise de Componentes Principais. Os ingredientes foram assim classificados: a) baixa atrato-palatabilidade = farelos de trigo, soja e algodão, farinha e raspa de mandioca, farinha de girassol e fubá de milho, b) média atrato-palatabilidade = levedura de cana-de-açúcar e glúten de milho e c) alta atrato-palatabilidade = ovo integral liofilizado, farinhas de crisálidas, peixes, carne e camarão. Ingestão de grânulos (manhã e tarde) e freqüência de visitas aos comedouros (tarde) foram consideradas as variáveis mais discriminatórias e freqüência de ejeção de grânulos e tempo gasto para aproximação dos comedouros (manhã e tarde), as menos discriminatórias. As respostas comportamentais dos peixes variaram de acordo com o ingrediente apresentado. A avaliação do grau de atrato-palatabilidade deve ser realizada considerando-se uma combinação de parâmetros.

Ultrastructure of the Lyonet's glands in larvae of Diatraea saccharalis Fabricius (Lepidoptera : Pyralidae)

The Lyonet's gland is found in Lepidoptera larvae, close to the excretory duct of the silk gland. The role played by this gland is still uncertain. This work aims to describe the ultrastructure of the Lyonet's bland in Diatraea saccharalis larvae, offering suggestions regarding its possible function. The insects were reared under laboratory-controlled conditions. The glands were conventionally prepared for transmission (TEM) and scanning (SEM) electron microscopy. SEM showed that Lyonet's glands are paired small structures located in the ventral side of the head. They are composed by clustered long cells resembling leaves. Under TEM observations, each cell is surrounded by a thin basal lamina and contains large stellate nucleus. The cytoplasm presents large and empty canaliculi with small microvilli. The basal plasma membrane forms numerous infoldings where numerous and well-developed mitochondria are concentrated. The cytoplasmic membrane system is poorly developed. Our ultrastructural results suggest that the Lyonet's gland in D. saccharalis larvae may be involved in the uptake of small molecules from the hemolymph no morphological evidences of macromolecules synthesis and secretion were noticed. The detection of nerve fibers in the gland suggest a neural control for the glandular cell function.