Pulvinus functional traits in relation to leaf movements: A light and transmission electron microscopy study of the vascular system

Previous studies on legume pulvini suggest that the vascular system plays an important role in the redistribution of ions and transmission of stimuli during leaf's movements. However, the number of anatomical and ultrastructural studies is limited to few species. The aim of this paper is to investigate the structure and cellular features of the pulvinus vascular system of nine legume species from Brazilian cerrado, looking for structural traits pointing to its participation in the leaf's movements. Samples were excised from the medial region of opened pulvinus of Bauhinia rufa, Copaifera langsdorffii, Senna rugosa (Caesalpinioideae), Andira humilis, Dalbergia miscolobium, Zornia dilphylla (Faboideae), Mimosa rixosa, Mimosa flexuosa and Stryphnodendron polyphyllum (Mimosoideae), and were prepared following light microscopy, transmission electron microscopy and histochemical standard techniques. The vascular system occupies a central position, comprises phloem and xylem and is delimited by a living sheath of septate fibers in all the species studied. This living cells sheath connects the cortex to the vascular tissues via numerous plasmodesmata. The absence of fibers and sclereids, the presence of phenolic idioblasts and the abundance and diversity of protein inclusions in the sieve tube members are remarkable features of the phloem. Pitted vessel elements, parenchyma cells with abundant cytoplasm and living fibriform elements characterize the xylem. The lack of lignified tissues and extensive symplastic continuity by plasmodesmata are remarkable features of the vascular system of pulvini of the all studied species. (c) 2007 Elsevier Ltd. All rights reserved.

Intracellular papillae of Actinocephalus (Eriocaulaceae-Poales) roots and their interaction with fungi: A light and transmission electron microscopy study

The genus Actinocephalus comprises 25 species and is restricted to Brazil, occurring mainly in the Espinhaco Mountains of Minas Gerais and Bahia States. Previous anatomical studies have reported the occurrence of intracellular papillae in the Actinocephalus roots, without dealing with their ultrastructure and function. The purpose of this paper is to investigate the structure, the composition and the probable function of the intracellular papillae of Actinocephalus roots, based on light microscopy, transmission electron microscopy and histochemical tests. The intracellular papillae occurred in all root tissues, from the rhizodermis to the vascular cylinder; they presented different forms and sizes and, ultrastructurally, they corresponded to material deposited between the cell wall and the plasma membrane. The histochemical tests carried out were positive for cellulose, pectin and callose. The intracellular papillae are responses of the plant cells to the interaction with fungi. They work as a physical barrier restricting fungal penetration, and they may also favor the supply of water and nutrients to the plant, since they increase root absorption surface. This might explain why the species of Actinocephalus are among the tallest Eriocaulaceae despite their reduced radicular system and the nutritional deficiency of the soil in which they grow. (C) 2006 Elsevier Ltd. All rights reserved.

The use of transmission electron microscopy and oocyte transfer to evaluate in vitro maturation of equine oocytes in different culture conditions

The current study evaluates the ability of equine oocytes matured in different conditions to undergo nuclear and cytoplasmic maturation.. After oocyte transfer, embryonic development was diagnosed at 1.5 and 90 days of gestation. For each group, immature oocytes obtained from slaughterhouse ovaries were matured in vitro (5 replicates). In experiment I, three different media were tested. HTF:BME, SOFaa, and TCM 199. In experiment 11, the HTF:BME was chosen as maturation medium containing pFSH, eFSH, or eFSH + eGH. Nuclear maturation was estimated after stripping the oocytes and staining with Hoechst 33342. The evaluation of cytoplasmic maturation was performed by transmission electron microscopy. For oocyte transfer, six non-cycling recipient mares were used, and 8 to 15 oocytes were transferred in each mare. In experiment I, the results showed no differences (P > .05) in nuclear maturation (MII) among experimental groups. The percentage of MII was 29.3 ( +/- 9.6), 23.4 ( +/- 8.4), and 13.5 ( +/- 12.4) for HTF:BME, SOF, and TCM, respectively. In experiment II, all media tested were efficient in inducing metaphase II. Also, no statistical differences (P > .05) were observed in percentages of nuclear maturation rates when porcine (37.1 +/- 22.4) or equine (25.8 +/- 8.2) FSH were used, or when eFSH + eGH was added to HTF:BME (29.4 +/- 12.3). The analysis of cytoplasmic morphology of oocytes cultured in TCM 199 and SOFaa showed signs of incomplete cytoplasmic maturation and premature cortical reaction. Meanwhile, oocytes cultured in HTF:BME medium presented cytoplasmic characteristics similar to those described by others for in vivo-matured oocytes. The addition of eFSH to the HTF:BME medium resulted in an improvement of cytoplasmic morphology. After oocyte transfer, two mares became pregnant, one from pFSH group and one from eFSH+eGH group. These results indicate that although in vitro matured equine oocytes are capable of fertilization and embryonic development, the percentage of competent oocytes is still low.

Cell enumeration and visualisation by transmission electron microscopy of Lactobacillus rhamnosus treated with cinnamon (Cinnamomum zeylanicum B.) essential oil

The use of essential oils (EOs) in functional foods containing probiotic microorganisms must consider the antimicrobial activity of these oils against beneficial bacteria such as Lactobacillus rhamnosus. This study aimed to evaluate the sensitivity of L. rhamnosus cultures treated with cinnamon EO through viable cell counts and visualisation by transmission electron microscopy. Cinnamon EO at a concentration of 0.04% had a bacteriostatic activity after 2 h of incubation. Although slight alterations were detected in the cell structure, this concentration was considered to be bactericidal, since it led to a significant reduction in cell numbers after 24 h. on the other hand, cinnamon EO at a 1.00% concentration decreased cell counts by 3 log units after 2 h incubation and no viable cell count was detected after 24 h. Transmission electron microscopy indicated that cells treated with 1.00% cinnamon EO were severely damaged and presented cell membrane disruption and cytoplasmic leakage.

Extruded vesicles of dioctadecyldimethylammonium bromide and chloride investigated by light scattering and cryogenic transmission electron microscopy

Combined dynamic and static light scattering (DLS, SLS) and cryogenic transmission electron microscopy (cryo-TEM) were used to investigate extruded cationic vesicles of dioctadecyldimethylammonium chloride and bromide (DODAX, X being Cl- or Br-). In salt-free dispersions the mean hydrodynamic diameter, D-h, and the weight average molecular weight, M-w, are larger for DODAB than for DODAC vesicles, and both D-h and M-w increase with the diameter (phi) of the extrusion filter. NaCl (NaBr) decreases (increases) the DODAB (DODAC) vesicle size, reflecting the general trend of DODAB to assemble as larger vesicles than DODAC. The polydispersity index is lower than 0.25, indicating the dispersions are rather polydisperse. Cryo-TEM micrographs show that the smaller vesicles are spherical while the larger ones are oblong or faceted, and the vesicle samples are fairly polydisperse in size and morphology. They also indicate that the vesicle size increases with phi and DODAB assembles as larger vesicles than DODAC. Lens-shaped vesicles were observed in the extruded preparations. Both light scattering and cryo-TEM indicate that the vesicle size is larger or smaller than phi when phi is smaller or larger than the optimal phi* approximate to 200 nm. (C) 2008 Elsevier B.V. All rights reserved.

Transmission electron microscopy for characterization of acrosomal damage after Percoll gradient centrifugation of cryopreserved bovine spermatozoa

The objective of this study was to characterize acrosomal ultrastructure following discontinuous Percoll gradient centrifugation of cryopreserved bovine sperm. Semen was collected from six bulls of different breeds and three ejaculates per bull were evaluated. Frozen semen samples were thawed and the acrosomal region of sperm cells was evaluated by transmission electron microscopy (TEM) before (n = 18) and after (n = 18) Percoll centrifugation. The evaluation of 20 sperm heads from each of the 36 samples analyzed ensured that a large number of cells were investigated. The data were subjected to analysis of variance at a level of significance of 5%. Percoll centrifugation reduced the percentage of sperm exhibiting normal acrosomes (from 61.77 to 30.24%), reduced the percentage of sperm presenting atypical acrosome reactions (from 28.38 to 4.84%) and increased the percentage of sperm exhibiting damage in the acrosome (from 6.14 to 64.26%). The percentage of sperm with typical acrosome reactions was not significantly different before (3.70%) and after (0.67%) centrifugation. TEM distinguished four different types of acrosomal status and enabled ultrastructural characterization of acrosomal injuries. The percentage of sperm exhibiting normal acrosomes decreased and damage in the acrosome was the most frequent acrosomal injury with the Percoll gradient centrifugation protocol utilized.

Experimental paracoccidioidomycosis in hamster: transmission electron microscopy of inoculation site lesion

Estudou-se sequencialmente, à microscopia eletrônica de transmissão, a interação entre Paracoccidioides brasiliensis (Pb) e células inflamatórias em hamsters inoculados por via intratesticular. Seis horas após inoculações havia predominância de neutrófilos, estando presentes algumas células mononucleares e eosinófilos. Os neutrófilos foram progressivamente substituídos por células mononucleares. Fungos viáveis apresentavam-se fagocitados ou circunscritos por células inflamatórias, geralmente com ampla interface hospedeiro-parasita. Fungos mortos ou degenerados eram acompanhados de interfase estreita. A camada externa da parede do Pb era às vezes quebrada quando em contacto com neutrófilos, em vários pontos, sendo os fragmentos dessa parede descamados e fagocitados. Células fúngicas pequenas com um único núcleo se relacionavam com ampla interfase enquanto as células maiores e multinucleadas apresentavam paredes irregulares, por vezes, contendo lomasoma e/ou estrutura semelhante à mielina. Diferentes padrões de interação do Pb com células do hospedeiro podem ser decorrentes do a fluxo de células inflamatórias funcionalmente diferentes ao local de inoculação ou à idade dos fungos ou ambos os fatores.

Study of ITO@PMMA composites by transmission electron microscopy

ITO nanowires were synthesized by carbothermal reduction process, using a co-evaporation method, and have controlled size, shape, and chemical composition. The electrical measurements of nanowires showed they have a resistance of about 102 Ω. In order to produce nanocomposites films, nanowires were dispersed in toluene using an ultrasonic cleaner, so the PMMA polymer was added, and the system was kept under agitation up to obtain a clear suspension. The PMMA polymer was filled with 1, 2, 5 and 10% in weight of nanowires, and the films were done by tape casting. The results showed that the electrical resistance of nanocomposites changed by over 7 orders of magnitude by increasing the amount of filler, and using 5 wt% of filler the composite resistance decreased from 1010 Ω to about 104 Ω, which means that percolation threshold of wires occurred at this concentration. This is an interesting result once for nanocomposites filled with ITO nanoparticles is necessary about 18% in weight to obtain percolation. The addition of filler up to 10 wt% decreased the resistance of the composite to 103 Ω, which is a value close to the resistance of wires. The composites were also analyzed by transmission electron microscopy (TEM), and the TEM results are in agreement with the electrical ones about percolation of nanowires. These results are promising once indicates that is possible to produce conductive and transparent in the visible range films by the addition of ITO nanowires in a polymeric matrix using a simple route. © 2011 Materials Research Society.

Spermatozoon ultrastructure and sperm cryopreservation of the Brazilian dry season spawner fish pirapitinga, Brycon nattereri

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Symbiotic bacteria and the structural specializations in the ileum of Cephalotes ants

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

ZnO architectures synthesized by a microwave-assisted hydrothermal method and their photoluminescence properties

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Net energy, protein and macrominerals requirements for 70 to 120 day old female rabbits

In order to determine the net energy, protein and macrominerals requirements of 70 to 120 day old, 52 female White New Zealand rabbits, weighing 1900g +/- 40g were used. At the beginning of the experimental period, 14 of the 52 young does were slaughtered and the 38 remaining animals were kept under two dietary management: ad libitum and restricted feeding. Slaughters were performed to determine each nutrient body content. The weight gain nutrient requirements depicted by the quantities of each nutrient stored into the body were obtained by applying the regression equation, which estimate the empty body nutrient content logarithm as a function of the empty body weight logarithm, as described by ARC (1980). By determining the heat production logarithm at the zero level of metabolizable energy intake, the maintenance net energy requirement was estimated to be 45.31 Kcal/day/Kg(0.75) the mean net energy. protein, calcium, phosphorous, sodium, magnesium and potassium requirements for each gram of weight gain per day were estimated to be, 2.51 Kcal, 0.21g, 0.02g, 0.005g, 0.001g, 0.0004g and 0.002g, respectively.

Segmental specializations in the Malpighian tubules of the fire ant Solenopsis saevissima Forel 1904 (Myrmicinae): an electron microscopical study

The Malpighian tubules of workers of the fire ant Solenopsis saevissima (Myrmicinae) were analyzed by scanning and transmission electron microscopy in order to determine their functional organization and association with the hindgut epithelium. The ants showed six Malpighian tubules with three segments morphologically and structurally different. The proximal segment was long and its cells showed abundant smooth endoplasmic reticulum and lipid droplets, which suggest their role in lipid secretion. The mid segment was long and undulated and it was composed by the cells that showed the typical features of ion transporting epithelia. The distal segment, short and flattened, adheres to the rectum wall. The cells of this segment showed the basal lamina fused to that of the rectum, it is probable that this part of the tubule may play a role in ion and water uptake from the feces. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Initial Development of Bovine Placentation (Bos indicus) from the Point of View of the Allantois and Amnion

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