Optimal single-embryo mass spectrometry fingerprinting

In pre-implantation embryos, lipids play key roles in determining viability, cryopreservation and implantation properties, but often their analysis is analytically challenging because of the few picograms of analytes present in each of them. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) allows obtaining individual phospholipid profiles of these microscopic organisms. This technique is sensitive enough to enable analysis of individual intact embryos and monitoring the changes in membrane lipid composition in the early stages of development serving as screening method for studies of biology and biotechnologies of reproduction. This article introduces an improved, more comprehensive MALDI-MS lipid fingerprinting approach that considerably increases the lipid information obtained from a single embryo. Using bovine embryos as a biological model, we have also tested optimal sample storage and handling conditions before the MALDI-MS analysis. Improved information at the molecular level is provided by the use of a binary matrix that enables phosphatidylcholines, sphingomyelins, phosphatidylserines, phosphatidylinositols and phosphoethanolamines to be detected via MALDI(±)-MS in both the positive and negative ion modes. An optimal MALDI-MS protocol for lipidomic monitoring of a single intact embryo is therefore reported with potential applications in human and animal reproduction, cell development and stem cell research. Copyright © 2013 John Wiley & Sons, Ltd. Copyright © 2013 John Wiley & Sons, Ltd.

Ecotoxicology of Nano-TiO2 An Evaluation of its Toxicity to Organisms of Aquatic Ecosystems

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Gross and microscopic observations on the lingual structure of the franciscana (Pontoporia blainvillei-gervais and d'orbigny, 1844)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Abundance of sedentary consumers and sessile organisms along the wave exposure gradient of subtropical rocky shores of the south-west Atlantic

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

Toxicity of the therapeutic potassium permanganate to tilapia Oreochromis niloticus and to non-target organisms Ceriodaphnia dubia (microcrustacean cladocera) and Pseudokirchneriella subcapitata (green microalgae)

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

Ovine leptospirosis in Brazil

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

Microscopic features of tick-bite lesions in anteaters and armadillos - Emas National Park and the Pantanal region of Brazil

The naturally occurring wildlife host associations between ticks and tick-borne pathogens found in the neotropics are poorly described. Understanding tick-bite lesions is important as these are the site of host reaction to and pathogen delivery by ticks. As part of a comprehensive study concerning established and emerging tick-host relationships. The present work describes some aspects of tick-bite lesions in anteaters and armadillos captured at the Emas National Park and the Pantanal region of Brazil. Biopsies were of skin were taken and examine. Tick feeding sites of all animals displayed an eosinophilic homogeneous mass, the cement cone, and, occasionally, a feeding cavity underneath the tick attachment site. At these locations the epidermis was usually thickened due to keratinocyte hyperplasia. The main dermal changes included tissue infiltration with a varying number of inflammatory cells, edema, hemorrhage. and vascular dilatation. Cellular infiltration of the dermis was predominantly composed of mononuclear cells, neutrophils. and eosinophils. Mast cells were also seen in both non-parasitized and parasitized skin but were found in higher numbers at perivascular sites and in parasitized skin. Basophils were not seen at tick attachment sites of anteaters or armadillos.

Molecular diagnosis of Anaplasmataceae organisms in dogs with clinical and microscopical signs of ehrlichiosis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Serologic evidence of equine granulocytic anaplasmosis in horses from central West Brazil

A Erliquiose é uma doença zoonótica causada por bactérias gram-negativas e intracelulares obrigatórias. A Anaplasmose Granulocítica Equina - AGE (anteriormente denominada Erliquiose Granulocítica Equina, EGE) é uma enfermidade sazonal, normalmente auto-limitante em equinos. No Brasil, existem poucos relatos deste agente erliquial, bem como de seus vetores naturais. Atualmente, veterinários têm levantado a suspeita de casos de AGE em equinos com sinais clínicos sugestivos de erliquiose e não responsivos ao tratamento para a piroplasmose equina. O objetivo do presente estudo foi identificar equinos expostos a A. phagocytophilum por meio de técnicas sorológicas e moleculares. Vinte amostras de sangue e soro de equinos da região Centro-oeste do Brasil foram avaliados por meio do exame microscópico de capa leucocitária, ensaio imunoenzimático indireto (ELISA), reação de imunofluorescência indireta (RIFI) e reação em cadeia da polimerase (nested PCR). Adicionalmente, o diagnóstico sorológico de Theileria equi pela RIFI e ELISA foram realizados, assim como o diagnóstico molecular pelo nPCR. Treze (65%) amostras de soro foram positivas para A. phagocytophilum pelo teste de ELISA, entretanto nenhum equino foi positivo pelo exame microscópico da capa leucocitária ou nPCR. Anticorpos IgG anti-T. equi foram detectados em 18 (90%) e 17 (85%) equinos pela RIFI e ELISA, respectivamente e o agente foi detectado em 9 (45%) animais pelo nPCR. Estes dados sugerem importante informação para o entendimento da ocorrência da AGE e piroplasmose equina no Centro-oeste do Brasil.

Laparoscopic ovum collection in sheep: Gross and microscopic evaluation of the ovary and influence on ooctye production

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

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.

Prevalence and macro- and microscopic lesions produced by Gasterophilus nasalis (Diptera : Oestridae) in the Botucatu Region, SP, Brazil

The objective of the present study was to investigate the prevalence of Gasterophilus nasalis larvae in Botucatu, the central west region of São Paulo State, Brazil, and to describe the lesions caused by the parasite. The climate of Botucatu is warm and rainy during the months of December through March and cool and dry during the months of May through August. The prevalence of G. nasalis was 16.84%, and the period of peak infestation corresponded to the months of cold and dry weather. The lesions detected at the sites of larval attachment were examined macro- and microscopically. Erosions and ulcerations of the gastric mucosa and proximal duodenum were the major macroscopic lesions detected. Microscopically, the lesion spectrum ranged from mild inflammatory reactions to extensive necrosis and ulceration. (C) 2001 Elsevier B.V. B.V. All rights reserved.

Microscopic evaluation of induced tooth movement in traumatized teeth: an experimental study in rats

The clinical management of orthodontic patients with dental trauma before or during the treatment is mainly founded on clinical experience, expert opinions, and individual case reports. It is proposed in the literature that teeth sustaining mild trauma with minor damage to the periodontium (e.g. subluxation) should be followed for a period of time before being subjected to orthodontic forces. A minimum period of 3 months has been proposed. In this study, we used an animal model to investigate whether shorter observation periods could be established in case of mild trauma. The periradicular region of rat molars was examined microscopically to determine the biological events of tooth movement started 15 and 30 days after intentional subluxation using an experimental method to induce dentoalveolar trauma. Thirty adult male Wistar rats were assigned to 6 groups (n = 5): Group 1 (control no trauma/orthodontic movement); Group 2: the animals received an orthodontic device and were sacrificed after 7 days; Groups 3 and 4: dentoalveolar trauma (subluxation) was experimentally induced by the application of an axial force of 900 cN on the occlusal surface of the maxillary right first molar, and the animals were sacrificed after 22 and 37 days, respectively; and Groups 5 and 6: 15 and 30 days, respectively, after force application, an orthodontic device was installed and the rats were sacrificed 7 days later. In G5 and G6, the periodontal ligament and pulp tissue were rich in cellular elements and blood vessels, the alveolar bone was preserved, and the root surface presented only very small areas of surface resorption (cementum), maintaining the characteristics of normality. In conclusion, the microscopic alterations in the gingival and periodontal tissues in response to an experimentally induced mild dentoalveolar trauma simulating subluxation were not sufficient to contraindicate starting the orthodontic movement 15 and 30 days after trauma.

Propolis as storage media for avulsed teeth: microscopic and morphometric analysis in rats

The maintenance of the avulsed teeth in appropriate media for preserving the cellular viability has been important for repairing the periodontal ligament and preventing the root resorption after tooth reimplantation. Propolis is a substance capable of preserving cellular viability. This study aimed to analyze the propolis substance as a storage media for maintaining the avulsed teeth, besides to determine the ideal time period for keeping the tooth inside it. Thus, 60 maxillary right central incisors of rats were extracted and divided into five groups. In groups I and II, teeth were kept in propolis for 60 min and 6 h, respectively; in group III, teeth were kept in milk for 6 h; in group IV, teeth were kept dry for 60 min; and in group V, they were immediately reimplanted. All teeth had their root canals filled with calcium hydroxide paste. Following, teeth were reimplanted in their sockets. After 15 and 60 days, animals were killed and the obtained samples were processed in laboratory for microscopic and morphometric analyzing. The results showed that the occurrence of inflammatory resorption, dental ankylosis and the formation of the connective tissue parallel to the root surface were similar among groups. It could be verified a greater occurrence of replacement resorption in group IV when comparing to other groups. In groups I and IV, the presence of periodontal ligament-like connective tissue was substantially smaller than the other groups. Regarding to the cementum amount over the root, it could be observed that this was present in smaller amount in groups I and IV. Group II was similar to groups III and IV. Therefore, according to the results of this study, the use of propolis as a storage media for maintaining avulsed teeth could be highlighted, and the 6-h period was more appropriate than the 60-min period.

Analysis of the healing process in delayed tooth replantation after root canal filling with calcium hydroxide, Sealapex and Endofill: a microscopic study in rats

The major concern in the therapeutics of tooth replantation refers to the occurrence of root resorption and different approaches have been proposed to prevent or treat these complications. The purpose of this study was to evaluate tissue response to delayed replantation of anterior rat teeth treated endodontically using calcium hydroxide, Sealapex, and Endofill without the placement of gutta-percha cones. Thirty rats had their right upper incisor extracted and maintained in dry storage for 60 min. After removal of the dental papilla, enamel organ, pulp tissue, and periodontal ligament remnants, the teeth were immersed in 2% sodium fluoride phosphate acidulated, pH 5.5, for 10 min. The root canals were dried with absorbent paper points and the teeth were assigned to three groups (n = 10) according to the filling material. Group I - calcium hydroxide and propyleneglycol paste, Group II - Sealapex, and Group III - Endofill. The sockets were irrigated with saline and the teeth were replanted. Replacement resorption, inflammatory resorption and ankylosis were observed in all groups. Although the occurrence of inflammatory resorption was less frequent in Group I, there were no statistically significant differences among the groups. It may be concluded that compared to the paste, filling the root canals with Sealapex and Endofill sealers without the placement of gutta-percha cones did not provide better results.