Scanning electron microscope evaluation of chlorhexidine gel and liquid associated with sodium hypochlorite cleaning on the root canal walls

Objective. The aim of this study was to evaluate, by scanning electronic microscopy (SEM), the cleaning of the root canal walls after instrumentation and irrigation with 2.5% sodium hypochlorite (NaOCl) associated with 2% chlorhexidine (CHX) gel or liquid, combined or not with 17% ethylenediamine tetra-acetic acid (EDTA).Study design. Sixty single-root human teeth were subjected to standardized root canal instrumentation with different irrigants (n = 10): G1) NaOCl + CHX liquid; G2) NaOCl + CHX liquid + EDTA + saline solution; G3) NaOCl + CHX gel; G4) NaOCl + CHX gel + EDTA + saline solution; G5) saline solution; G6) saline solution + EDTA. After instrumentation, the teeth were prepared for SEM analysis (x500 and x2,000) to evaluate the cleaning of the cervical, middle, and apical thirds. The area analyzed was quantified according to the percentage of open and closed tubules, and data were statistically analyzed by analysis of variance and Tukey tests (P = .05).Results. The number of open tubules was highest in G4 in all root thirds, showing statistically significant difference from G1, G2, and G5 (P < .05). G1 presented higher quantity of closed tubules significant than G2.Conclusion. Irrigation with NaOCl and CHX gel followed by EDTA and saline solution produced greater cleaning of the root canal walls. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010;110:e82-e87)

OBSERVATIONS on ETCHED ENAMEL IN NON-ERUPTED DECIDUOUS MOLARS: A SCANNING ELECTRON MICROSCOPIC STUDY

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

PRISMLESS ENAMEL IN HUMAN NON-ERUPTED DECIDUOUS MOLAR TEETH: A SCANNING ELECTRON MICROSCOPIC STUDY

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Light microscopic evaluation and scanning electron microscopic analysis of horse eyes following deep anterior lamellar keratectomy

Objective To describe the technique of deep anterior lamellar keratoplasty (DALK) with Descemet's membrane (DM) exposure in horse eyes. Also, to compare the efficacy and safety of viscodissection and big-bubble techniques for DALK. Animals studied Thirty-four ex vivo horse eyes. Procedure Deep anterior lamellar keratoplasty was performed in 34 ex vivo horse eyes. Two groups (Group V - viscodissection - 2% sodium hyaluronate; Group A - air - big-bubble) of 17 eyes were studied. Other than the substance used, the surgical technique was similar for both groups. Nonperforated eyes were submitted for light microscopic histologic evaluation and scanning electron microscopic (SEM) analysis. Results Group V - Perforations occurred in 18% of the eyes during surgery. Light microscopy revealed exposure of DM in 28% of the eyes with mean thickness of the remaining stroma being 70.4μm. Group A - Perforations occurred in 42% of the eyes. Light microscopy revealed exposure of DM in 60% of the eyes with mean thickness of the remaining stroma being 23.3μm. No significant differences in safety, efficacy and thickness of the remaining stroma (including all eyes or excluding those with DM exposure) were observed. SEM of the surgical site revealed a more even surface in those eyes with DM exposure compared to eyes with thicker remaining stroma in both groups. Conclusions We describe two DALK techniques (viscodissection and big-bubble) for use in horses. No significant differences in safety, efficacy and thickness of the remaining stroma were observed. However, a nonsignificant trend toward the big-bubble technique being more efficacious but less safe was observed. © 2012 American College of Veterinary Ophthalmologists.

Effect of Er:YAG laser and diamond drill on hybrid layer morphology obtained with self-etch adhesive: analysis by SEM and confocal laser scanning microscopy (CLSM)

This study aimed to evaluate the effect of Er:YAG (L) and diamond drills (DD) on: 1) the microshear bond strength (MPa); 2) the adhesive interface of two-step (TS) – Adper Scotchbond Multipurpose and one-step (OS) adhesives – Adper EasyOne, both from 3M ESPE. Material and methods: According to the preparation condition and adhesives, the samples were divided into four groups: DD_TS (control); DD_OS; L_TS and L_OS. 60 bovine incisors were randomly divided into experimental and groups: 40 for microshear bond strength (n = 10) and 20 for the adhesive interface morphology [6 to measure the thickness of the hybrid layer (HL) and length of tags (t) by CLSM (n = 3); 12 to the adhesive interface morphology by SEM (n = 3) and 2 to illustrate the effect of the instruments on dentine by SEM (n = 1)]. To conduct the microshear bond strength test, four cylinders (0.7 mm in diameter and 1 mm in height with area of adhesion of 0.38 mm) were constructed with resin composite (Filtek Z350 XT – 3M ESPE) on each dentin surface treated by either L or DD and after adhesives application. Microshear bond strength was performed in universal testing machine (EMIC 2000) with load cell of 500 kgf and a crosshead speed of 0.5 mm / min. Adhesive interface was characterized by thickness of hybrid layer (HL) and length of tags (t) in nm, with the aid of UTHSCSA ImageTool software. Results: Microshear bond strength values were: L_TS 34.10 ± 19.07, DD_TS 24.26 ± 9.35, L_OS 33.18 ± 12.46, DD_OS 21.24 ± 13.96. Two-way ANOVA resulted in statistically significant differences only for instruments (p = 0.047). Mann-Whitney identified the instruments which determined significant differences for HL thickness and tag length (t). Concerning to the adhesive types, these differences were only observed for (t). Conclusion: It can be concluded that 1) laser Er:YAG results in higher microshear bond strength values regardless of the adhesive system (TS and OS); 2) the tags did not significant affect the microshear bond strength; 3) the adhesive interface was affected by both the instruments for cavity preparation and the type of adhesive system used.

Identification of new isolates of Bacillus thuringiensis using rep-PCR products and delta-endotoxin electron microscopy

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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.

Chemical and microbiological evaluation of the internal surfaces of aluminum tubes both unlined and lined with epoxi resin by means of the stereoscope and scanning electron microscope

A superfície interna das bisnagas fabricadas com alumínio não revestido e revestido com resina epóxi, utilizadas para acondicionar cremes, pomadas, géis, etc., foram avaliadas quimicamente e por métodos microbiológicos correlacionados com a aderência de microrganismos. A prova da porosidade e da resistência à remoção da resina foi observada por meio do microscópio eletrônico de varredura (Topcon FM300) e estereoscópio Leica (MZ12) acoplado a Sistema de Digitalização de Imagens. Para avaliar a ação dos microrganismos foram utilizados corpos-de-prova esterilizados (discos de 10mm de diâmetro), imersos em caldo Mueller Hinton (Difco) e colocados em tubos de polipropileno com tampa de rosca (Corning). Foram inoculados tubos com meio de cultura para cada uma das suspensões bacterianas (10(9)UFC/mL) de Streptococcus agalactiae, Staphylococcus aureus, Acinetobacter lwoffii e Candida albicans, incubados a 37°C, sob agitação constante durante 12 dias. O meio de cultura era trocado a cada 3 dias. Após esse período, os corpos-de prova foram removidos, processados e observados em microscópio eletrônico de varredura JEOL-JSM (T330A). A observação por meio do microscopio eletrônico de varredura mostrou a aderência e a formação de biofilme sobre a superfície de alumínio não revestido e revestido com resina epóxi.

Is Reinke's Edema a Precancerous Lesion? Histological and Electron Microscopic Aspects

The objectives of this study were to evaluate morphologic alterations and precancerous lesions in Reinke's edema. Patients included were 54 smokers with bilateral Reinke's edema submitted to surgery in the Otolaryngology Department, Botucatu Medical School, São Paulo State University, Brazil, between 2002 and 2006. All specimens were evaluated by light microscopy and five contralateral lesions were also evaluated by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The main histological alterations were edema (100%), inflammation (81.48%), basal membrane (bm) thickening (77.77%), and vessel proliferation (75.92%). Epithelium alterations were classified as grade 0 (11.11%), grade 1 (70.37%), grade 2 (14.81%), and grade 3 (3.70%). In the case included in grade 3 classification, microinvasive carcinoma was detected. SEM showed epithelial surface with some cellular desquamation, few microridges, and a polished and impermeable surface aspect. TEM showed epithelial hyperplasia, basal cells with different sizes, widening of the intercellular spaces, abnormal desmosome architecture, thickening of the bm, some electron-dense vesicles, and points of interruption. The morphological alterations presented in this study are not specific to Reinke's edema but this lesion can be the site of different grades of dysplasia and carcinoma, justifying the importance of periodic laryngeal endoscopic exams and meticulous histological analysis.

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.

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.