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.

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.

Development of secondary sexual characters in the seabob shrimp Xiphopenaeus kroyeri (Heller 1862) (Crustacea, Decapoda, Penaeidae): a scanning electron microscope study

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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.

Annexin 1 localisation in tissue eosinophils as detected by electron microscopy

BACKGROUND: Human and rodent leukocytes express high levels of the glucocorticoid-inducible protein annexin 1 ( ANXA1) ( previously referred to as lipocortin 1). Neutrophils and monocytes have abundant ANXA1 levels.Aim: We have investigated, for the first time, ANXA1 ultrastructural expression in rat eosinophils and compared it with that of extravasated neutrophils. The effect of inflammation ( carrageenin peritonitis) was also monitored.Methods: Electron microscopy was used to define the sub-cellular localisation of ANXA1 in rat eosinophils and neutrophils extravasated in the mesenteric tissue. A pair of antibodies raised against the ANXA1 N-terminus (i.e. able to recognise intact ANXA1, termed LCPS1) or the whole protein ( termed LCS3) was used to perform the ultrastructural analysis.Results: the majority of ANXA1 was localised in the eosinophil cytosol (similar to 60%) and nucleus (30-40%), whereas a small percentage was found on the plasma membrane (< 10%). Within the cytosol, the protein was equally distributed in the matrix and in the granules, including those containing the typical crystalloid. The two anti-ANXA1 antibodies gave similar results, with the exception that LCPS1 gave a lower degree of immunoreactivity in the plasma membrane. Inflammation (i.e. carrageenin injection) produced a modest increase in eosinophil-associated ANXA1 reactivity ( significant only in the cytoplasm compartment). Extravasated neutrophils, used for comparative purposes, displayed a much higher degree of immunoreactivity for the protein.Conclusion: We describe for the first time ANXA1 distribution in rat eosinophil by ultrastructural analysis, and report a different protein mobilisation from extravasated neutrophils, at least in this acute model of peritonitis.

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.

Microbiological or Chemical Models of Enamel Secondary Caries Compared by Polarized-Light Microscopy and Energy Dispersive X-ray Spectroscopy

Different secondary caries models may present different results. The purpose of this study was to compare different in vitro secondary caries models, evaluating the obtained results by polarized-light microscopy (PLM), scanning electron microscopy (SEM), and energy dispersive X-ray spectroscopy (EDS). Standardized human enamel specimens (n = 12) restored with different materials (Z250 conventional composite resin-CRZ, Freedom polyacid-modified composite resin-CRF, Vitremer resin-modified glass-ionomer-GIV, and Fuji IX conventional glass-ionomer cement-GIF) were submitted to microbiological (MM) or chemical caries models (CM). The control group was not submitted to any caries model. For MM, specimens were immersed firstly in sucrose broth inoculated with Streptococcus mutans ATCC 35688, incubated at 37 degrees C/5% CO(2) for 14 days and then in remineralizing solution for 14 days. For CM, specimens were submitted to chemical pH-cycling. Specimens were ground, submitted to PLM and then were dehydrated, gold-sputtered and submitted to SEM and EDS. Results were statistically analyzed by Kruskall-Wallis and Student-Newman-Keuls tests (alpha = 0.05). No differences between in vitro caries models were found. Morphological differences in enamel demineralization were found between composite resin and polyacid-modified composite resin (CRZ and CRF) and between the resin-modified glass-ionomer and the glass-ionomer cement (GIF and GIV). GIF showed higher calcium concentration and less demineralization, differing from the other materials. In conclusion, the glass-ionomer cement showed less caries formation under both in vitro caries models evaluated. (C) 2009 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 90B: 635-640, 2009

Ability of different restorative materials to prevent in situ secondary caries: analysis by polarized light-microscopy and energy-dispersive X-ray

Secondary caries is the main cause of direct restoration replacement. The purpose of this study was to analyze enamel adjacent to different restorative materials after in situ cariogenic challenge using polarized-light microscopy (PLM), scanning electron microscopy (SEM) and energy-dispersive X-ray analysis (EDS). Twelve volunteers, with a low level of dental plaque, a low level of mutans streptococci, and normal salivary flow, wore removable palatal acrylic appliances containing enamel specimens restored with Z250 composite, Freedom composite, Fuji IX glass-ionomer cement, or Vitremer resin-modified glass-ionomer for 14 days. Volunteers dripped one drop of 20% sucrose solution (n = 10) or distilled water (control group) onto each specimen 8 times per day. Specimens were removed from the appliances and submitted to PLM for examination of the lesion area (in mm(2)), followed by dehydration, gold-sputtering, and submission to SEM and EDS. The calcium (Ca) and phosphorus (P) contents were evaluated in weight per cent (%wt). Differences were found between Z250 and Vitremer, and between Z250 and FujiIX, when analyzed using PLM. Energy-dispersive X-ray analysis results showed differences between the studied materials regarding Ca %wt. In conclusion, enamel adjacent to glass-ionomer cement presented a higher Ca %wt, but this material did not completely prevent enamel secondary caries under in situ cariogenic challenge.

Removal of metal ions from aqueous solution by chelating polymeric hydrogel

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

Anatomical characterization and chemical profile of Laguncularia racemosa (L.) Gaertn leaf blades, from impacted and non-impacted mangroves of the São Paulo Coast

Anatomical characterization and chemical profile of Laguncularia racemosa (L.) Gaertn leaf blades, from impacted and non-impacted mangroves of the São Paulo Coast. Several sources of environmental impact have caused great damage to the flora in mangroves. Therefore, this study aimed to analyze the anatomy and chemical composition of Laguncularia racemosa (L.) Gaertn leaf blades, from impacted (Municipality of Santos) and non-impacted mangroves (Municipality of Peruibe) of the São Paulo Coast. For the leaf anatomy studies, light microscopy and scanning electron microscopy (SEM) were used. For chemical analysis, high performance liquid chromatography (HPLC) was conducted. The biometry revealed leaf tissues of plants from impacted mangroves with a less thick epidermis and mesophyll and higher width of the central vascular beam. A smaller number of stomata and higher amounts of daises in these leaves were also observed. Results were confirmed by the Student t-test (alpha = 5%). The chemical profiles revealed a higher ammount of secondary metabolites in leaves from the impacted environment.

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.

Scanning electron microscopic study of the cleaning ability of chlorhexidine as a root-canal irrigant

Aim To evaluate in vitro the cleaning of root-canal walls after irrigation with different irrigants.Methodology A total of 36 recently extracted human teeth were divided into four experimental groups according to the irrigating solution used: saline; 2% chlorhexidine; 2.5% sodium hypochlorite; and 2.5% sodium hypochlorite + EDTA. The cleaning of the apical, middle and coronal thirds of the root canals was evaluated by scanning electron microscope examination using a 4-point scoring system.Results the best cleaning was obtained using 2.5% sodium hypochlorite and EDTA, followed by 2.5% sodium hypochlorite only (P < 0.05), whose cleaning was similar to chlorhexidine only in the cervical third. Cleaning by saline and 2% chlorhexidine was worse than the other two groups and was similar in all thirds. Better cleaning was found in the cervical and middle thirds for all groups with the worst results in the apical third.Conclusions the apical third of the root canals was not cleaned as well as the middle and coronal thirds. Cleaning by chlorhexidine and saline was inferior compared to the cleaning by sodium hypochlorite with and without EDTA.

Coating residual stress effects on fatigue performance of 7050-T7451 aluminum alloy

The tendency of the aircraft industry is to enhance customer value by improving performance and reducing environmental impact. In view of availability, aluminum alloys have a historically tendency to faster insertion due to their lower manufacturing and operated production infrastructure. In landing gear components, wear and corrosion control of many components is accomplished by surface treatments of chrome electroplating on steel or anodizing of aluminum. One of the most interesting environmentally safer and cleaner alternatives for the replacement of hard chrome plating or anodizing is tungsten carbide thermal spray coating, applied by the high velocity oxy fuel (HVOF) process. However, it was observed that residual stresses originating from these coatings reduce the fatigue strength of a component.An effective process as shot peening treatment, considered to improve the fatigue strength, pushes the crack sources beneath the surface in most of medium and high cycle cases, due to the compressive residual stress field induced. The objective of this research is to evaluate a tungsten carbide cobalt (WC-Co) coating applied by the high velocity oxy fuel (HVOF) process, used to replace anodizing. Anodic films were grown on 7050-T7451 aluminum alloy by sulfuric acid anodizing, chromic acid anodizing and hard anodizing. The influence on axial fatigue strength of anodic films grown on the aluminum alloy surface is to degrade the stress-life performance of the base material. Three groups of specimens were prepared and tested in axial fatigue to obtain S-N curves: base material, base material coated by HVOF and base material shot peened and coated.Experimental results revealed increase in the fatigue strength of Al 7050-T7451 alloy associated with the WC 17% Co coating. on the other hand, a reduction in fatigue life occurred in the shot peened and coated condition. Scanning electron microscopy technique and optical microscopy were used to observe crack origin sites, thickness and coating/substrate adhesion. (c) 2007 Elsevier B.V. All rights reserved.

Characterization of the cellular component of polymorphous low-grade adenocarcinoma by immunohistochemistry and electron microscopy

In order to characterize the cellular component of the polymorphous low-grade adenocarcinoma (PLGA) of the salivary gland, a morphological and immunohistochemical study was carried out. Thirty cases of PLGA were studied by light microscopy and immunohistochemistry and five cases by transmission electron microscopy (TEM). The expression of cytokeratins (CKs) 7,8,10,13,14,18,19, vimentin and muscle-specific actin (MSA) was investigated through the streptavidin-biotin method. The majority of tumor cells stained for vimentin, CKs 8,18 and 7. CK 14 was positive in most cells of the papillary and trabecular sub-types. Although the expression of CKs 8,18 and 14 varied among the tumors sub-types, a straight relationship between each histologic pattern and the CK expression could not be delineated. MSA was reactive in only three tumors while CKs 10 and 13 were not detected in any tumor studied. The absence of MSA and the expression of CKs 8,18 and 7, in most of the tumor cells, lead to the hypothesis that myoepithelial cells are not the major cellular component of the PLGA. TEM revealed cells exhibiting microvilli and variable amounts of secretory granules, some of them suggesting an excretory activity. The presence of CKs 8, 18 and 7, added to the secretory granules, indicates that PLGA originates from cells located at the acinar-intercalated duct junction. (C) 1999 Elsevier B.V. Ltd. All rights reserved.