Impact of different silkworm dietary supplements on its silk performance

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Nephila clavipes e Nephila sexpunctata (Araneae, Araneomorphae, Nephilidae): cariótipo, heterocromatina constitutiva e região organizadora de nucléolo

As espécies de Nephila estão incluídas na família Nephilidae que pertence ao grupo das aranhas Entelegynae, o qual é considerado filogeneticamente derivado em relação aos outros grupos da ordem Araneae. Análises citogenéticas realizadas nas aranhas Entelegynae com técnicas de coloração convencional, têm mostrado que a maioria das espécies possui uniformidade cariotípica principalmente em relação a morfologia telo-acrocêntrica dos cromossomos e sistema cromossômico sexual (SCS) do tipo X1X20/X1X1X2X2. Além disso, estas análises têm demonstrado que algumas famílias de Entelegynae também possuem uniformidade cariotípica concernente ao número diplóide de cromossomos na maioria das suas espécies. Por outro lado, o emprego adicional de técnicas de coloração diferencial de regiões cromossômicas específicas em alguns representantes de Entelegynae têm revelado características que podem ser usadas para determinar os mecanismos envolvidos na evolução cromossômica e na diferenciação cariotípica de espécies relacionadas. Contudo, poucas espécies de Entelegynae tiveram seus cariótipos analisados com técnicas de coloração diferencial. O Brasil possui aproximadamente 4.000 espécies de Araneae descritas taxonomicamente; entretanto apenas cerca de 20 destas foram analisadas do ponto de vista citogenético. Considerando estas informações, o objetivo deste trabalho foi investigar o cariótipo de duas espécies de Nephila, Nephila clavipes e Nephila sexpunctata, com técnicas de coloração convencional e diferencial para determinar o número diplóide, a morfologia cromossômica, o tipo de SCS, e o padrão de distribuição da heterocromatina constitutiva (bandas C) e das regiões organizadoras de nucléolo (RONs) ativas, e comparar os dados obtidos com aqueles de espécies relacionadas, para estabelecer os mecanismos de evolução cromossômica...(Resumo completo, clicar acesso eletrônico abaixo)

Preparation and characterization of a bacterial cellulose/silk fibroin sponge scaffold for tissue regeneration

Bacterial cellulose (BC) and silk fibroin (SF) are natural biopolymers successfully applied in tissue engineering and biomedical fields. In this work nanocomposites based on BC and SF were prepared and characterized by scanning electron microscopy (SEM), infrared spectroscopy (FT-IR), X-ray diffraction (XRD) and thermogravimetric analysis (TGA). In addition, the investigation of cytocompatibility was done by MTT, XTT and Trypan Blue dye technique. Cellular adhesion and proliferation were detected additionally. The evaluation of genotoxicity was realized by micronucleus assay. In vitro tests showed that the material is non-cytotoxic or genotoxic. SEM images revealed a greater number of cells attached at the BC/SF:50% scaffold surface than the pure BC one, suggesting that the presence of fibroin improved cell attachment. This could be related to the SF amino acid sequence that acts as cell receptors facilitating cell adhesion and growth. Consequently, BC/SF:50% scaffolds configured an excellent option in bioengineering depicting its potential for tissue regeneration and cultivation of cells on nanocomposites.

Compositional nutrient diagnosis of corn using the Mahalanobis distance as nutrient imbalance index

Parent, L. E., Natale, W. and Ziadi, N. 2009. Compositional nutrient diagnosis of corn using the Mahalanobis distance as nutrient imbalance index. Can. J. Soil Sci. 89: 383-390. Compositional nutrient diagnosis (CND) provides a plant nutrient imbalance index (CND - r(2)) with assumed chi(2) distribution. The Mahalanobis distance D(2), which detects outliers in compositional data sets, also has a chi(2) distribution. The objective of this paper was to compare D(2) and CND - r(2) nutrient imbalance indexes in corn (Zea mays L.). We measured grain yield as well as N, P, K, Ca, Mg, Cu, Fe, Mn, and Zn concentrations in the ear leaf at silk stage for 210 calibration sites in the St. Lawrence Lowlands [2300-2700 corn thermal units (CTU)] as well as 30 phosphorus (2300-2700 CTU; 10 sites) and 10 nitrogen (1900-2100 CTU; one site) replicated fertilizer treatments for validation. We derived CND norms as mean, standard deviation, and the inverse covariance matrix of centred log ratios (clr) for high yielding specimens (>= 9.0 Mg grain ha(-1) at 150 g H(2)O kg(-1) moisture content) in the 2300-2700 CTU zone. Using chi(2) = 17 (P < 0.05) with nine degrees of freedom (i.e., nine nutrients) as a rejection criterion for outliers and a yield threshold of 8.6 Mg ha(-1) after Cate-Nelson partitioning between low- and high-yielders in the P validation data set, D(2) misclassified two specimens compared with nine for CND -r(2). The D(2) classification was not significantly different from a chi(2) classification (P > 0.05), but the CND - r(2) classification differed significantly from chi(2) or D(2) (P < 0.001). A threshold value for nutrient imbalance could thus be derived probabilistically for conducting D(2) diagnosis, while the CND - r(2) nutrient imbalance threshold must be calibrated using fertilizer trials. In the proposed CND - D(2) procedure, D(2) is first computed to classify the specimen as possible outlier. Thereafter, nutrient indices are ranked in their order of limitation. The D(2) norms appeared less effective in the 1900-2100 CTU zone.

Efeitos de três análogos do hormônio juvenil aplicados em bicho-da-seda, Bombyx mori L. (Lepidoptera: Bombycidae)

Análogos do hormônio juvenil (AHJ) têm sido testados como reguladores de crescimento de Bombyx mori L., com vistas ao incremento da produção de casulos. Esses produtos, quando aplicados em doses adequadas, prolongam o período larval. Os efeitos da aplicação de AHJ podem variar com o produto, a linhagem e o momento de aplicação. Este estudo teve por objetivo avaliar o efeito da aplicação tópica, em diferentes momentos, de três análogos do hormônio juvenil sobre o crescimento dos insetos e a produção de seda da linhagem brasileira C115xN108 do bicho-da-seda. Os produtos piriproxifem, metoprene e fenoxicarbe foram aplicados em larvas do bicho-da-seda a 24h, 48h, 72h e 96h após o início do quinto ínstar (HQI) e seus efeitos sobre características biológicas larvais e adultas e a produção de seda foram avaliados. Independentemente do momento de aplicação, os três produtos afetaram todos os parâmetros avaliados. A reserva de nutrientes acumulada durante o período larval prolongado dos insetos tratados foi destinada ao crescimento das glândulas sericígenas e, também, convertida em peso corporal, com conseqüentes incrementos na produção de seda. Aplicações entre 48 e 72 HQI provocaram os maiores incrementos na biomassa e na produção de casulos de B. mori. Variações no momento de aplicação de 24h e 96h não influenciaram a emergência ou a oviposição de adultos de B. mori, nem a viabilidade dos ovos.

Hypertension favors the inflammatory process in rats with experimentally induced periodontitis

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

Histometric analysis of ligature-induced periodontitis in rats: a comparison of histological section planes

The purpose of this study was to analyze the histometry of ligature-induced periodontitis in rats at different histological section depths. Sixteen male adult Wistar rats were randomly assigned to two groups: ligature and control. In the ligature group, rats received a sterile 4/0 silk ligature around the maxillary right 2nd molar. Thirty serial sections containing the 1st and 2nd molars, in which the coronal and root pulp, cementoenamel junction (CEJ) in the mesial side of the 2nd molar, interproximal alveolar bone and connective fiber attachment were clearly visible, were selected for histometric analysis. The histological sections were clustered in groups of 10 sections corresponding the buccal (B), central (C) and lingual (L) regions of the of periodontal tissue samples. The distance between the CEJ in the mesial side of the 2nd molar and the attached periodontal ligament fibers (CEJ-PL) as well as the distance between the CEJ and the alveolar bone crest (CEJ-BC) were determined. From CEJ-PL and CEJ-BC distances measured for each specimen, the measurements obtained in the B, L and C regions were recorded individually and together. Data were submitted to statistical analysis. Significant differences (p<0.001) were observed between the control and ligature groups regarding CEJ-PL (0.05 mm and 0.26 mm, respectively) and CEJ-BC (0.47 mm and 0.77 mm, respectively) measurements. Regarding the depth of the buccal, central and lingual planes, the means of CEJ-PL and CEJ-BC of both groups showed no statistically significant differences (p>0.05). In conclusion, the selection of 10 serial sections of the central region of periodontal tissue samples at any depth can be considered as representative for the evaluation of periodontal ligament fiber attachment and bone loss in ligature-induced periodontitis in rats.

The influence of chronic stress imposed on pregnant rats on the induced bone loss in their adult offspring

Background and objective: Stress during pregnancy may alter offspring susceptibility to diseases during adulthood. In the present study, female Lewis rats were subjected to chronic stress during the gestational period, and the effect of this stress was evaluated histometrically on the progression of ligature-induced bone loss in their adult offspring.Material and methods: After confirming pregnancy, half of the pregnant rats were randomly designated as control animals (no stress regimen was imposed), and the other half was submitted to a chronic stress model (immobilization at cold temperature) between the 7th and the 18th gestational day. After birth, 12 male rats delivered by stressed mothers - Group 1 (G1) - and 12 male rats delivered by non-stressed mothers - Group 2 (G2) - were selected. When birthed rats reached 250 g of body weight, a silk ligature was placed around their maxillary right second molar in order to induce bone loss. The non-ligated left side served as a control. Sixty days later, these animals were sacrificed by anaesthetic overdose. After routine laboratorial processing, images of the histological sections were digitized and submitted for histometric measurement using two parameters: histological attachment loss and bone loss.Results: on the ligated side, G1 presented with greater histological attachment and bone loss than G2 (p < 0.05). on the non-ligated control side, neither of the groups presented with alterations in these parameters (p > 0.05).Conclusion: The chronic stress regimen imposed on pregnant rats produced a greater progression of ligature-induced bone loss in their adult offspring. (C) 0 2011 Elsevier Ltd. All rights reserved.

Adesivos à base de cianoacrilato para síntese de tecido mole

FUNDAMENTOS - Adesivos teciduais têm sido muito usados para síntese de ferida, em função de ser um método indolor, rápido e de fácil execução. OBJETIVOS -Analisar e comparar compatibilidade dos adesivos, etil- cianoacrilato (Super Bonder) e butilcianoacrilato (Histoacryl), e a reparação de incisões em dorso de ratos entre o fio de sutura e os respectivos adesivos. MÉTODOS - Foram usados 15 ratos. Realizaram-se duas lojas cirúrgicas no dorso. em cada uma, foi implantado um tubo de polietileno (10mm x 1mm), os quais foram preenchidos com os adesivos Super Bonder (lado direito) e Histoacryl (lado esquerdo). As incisões, do lado esquerdo, foram coaptadas com Super Bonder, e as do lado direito, com Histoacryl. Uma incisão mediana, entre as duas incisões,foi realizada e suturada com fio de seda. Os animais foram mortos, depois de 7(sete), 35(trinta) e 120 (cento e vinte dia) dias. RESULTADOS: Os adesivos usados, no presente estudo, não promoveram reação inflamatória, quando usados para síntese das incisões. Porém, estes adesivos, quando implantados no subcutâneo, promoveram reação inflamatória até 120 (cento e vinte dia) dias, no entanto, a reação é mais intensa com Histoacryl. CONCLUSÕES: Super Bonder e Histoacryl permitem o processo cicatricial dos tecidos incisados; facilitam a sutura das incisões. Desta forma, estes podem ser utilizados para sínteses de feridas, lacerações ou incisões cutâneas.

In situ localization of heat-shock proteins and cell death labelling in the salivary gland of acaricide-treated honeybee larvae

The effects of the acaricides, rotenone and oxalic acid (OA) on salivary glands of honeybee larvae were evaluated. Immunohistochemical methods were used to detect cell death and heat-shock protein (HSP70 and 90) localizations. Heat-shock proteins (HSP70 and 90) were localized in the cytoplasm and/or the nuclei of secretory gland cells, both under stress and in normal conditions. In rotenone-treated larvae, there were no changes in the normal level of cell death and also there were no morphological alterations in the secretory cells. In the larvae treated with oxalic acid, the salivary gland showed varying degrees of morphological cellular alteration and an increase in the cell death level. The present data suggest that stress-induced HSP70 might have an antiapoptotic effect while the stress-induced HSP90 might have a chaperone function in the larval salivary glands.

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.