Viability and cell cycle analysis of equine fibroblasts cultured in vitro

This experiment aimed to study equine fibroblasts in culture analyzing and the cell cycle and viability of cells pre- and post-freezing. Skin fragments were obtained from 6 horses and cultured in DMEM high glucose + 10% FCS in 5% CO(2) until the beginning of confluence. Two passages were performed before freezing. Cells subjected to serum starvation (0.5% FCS) were analyzed for viability and cell cycle at 24, 48, 72, 96, 120, 144 and 168 h of culture. For the confluent groups, cells were analyzed at the moment they achieved confluence. Cellular viability was assisted with Hoescht 33342 and propidium iodide. The analysis of apoptosis/necrosis and cell cycle was performed using a flow cytometer (FACS Calibur BD(A (R))) after staining the cells with annexin V and propidium iodide. Both optical microscopy and flow cytometry confirmed that cellular viability was similar for serum starvation and confluent groups (average 84%). Similarly, both methods were efficient to synchronize the cell cycle before freezing. However, after thawing, serum starvation, for more than 24 h, was superior to culture for synchronizing cells in G0/G1 (69% x 90%). The results of this experiment indicate that equine fibroblasts can be efficiently cultured after thawing.

Influence of Different Base Thicknesses on Maxillary Complete Denture Processing: Linear and Angular Graphic Analysis on the Movement of Artificial Teeth

Objective: The purpose of this study was to compare the dental movement that occurs during the processing of maxillary complete dentures with 3 different base thicknesses, using 2 investment methods, and microwave polymerization.Methods: A sample of 42 denture models was randomly divided into 6 groups (n = 7), with base thicknesses of 1.25, 2.50, and 3.75 mm and gypsum or silicone flask investment. Points were demarcated on the distal surface of the second molars and on the back of the gypsum cast at the alveolar ridge level to allow linear and angular measurement using AutoCAD software. The data were subjected to analysis of variance with double factor, Tukey test and Fisher (post hoc).Results: Angular analysis of the varying methods and their interactions generated a statistical difference (P = 0.023) when the magnitudes of molar inclination were compared. Tooth movement was greater for thin-based prostheses, 1.25 mm (-0.234), versus thick 3.75 mm (0.2395), with antagonistic behavior. Prosthesis investment with silicone (0.053) showed greater vertical change compared with the gypsum investment (0.032). There was a difference between the point of analysis, demonstrating that the changes were not symmetric.Conclusions: All groups evaluated showed change in the position of artificial teeth after processing. The complete denture with a thin base (1.25 mm) and silicone investment showed the worst results, whereas intermediate thickness (2.50 mm) was demonstrated to be ideal for the denture base.

Correlation between morphology and function of the upper lip: a longitudinal evaluation

In order to evaluate the relationship between the morphology of the upper lip and muscle activity in a sample of 38 subjects (17 males and 21 females) with Angle Class II division 1 malocclusions, cephalometric and electromyographic analyses were conducted. The sample was subdivided into either predominantly nose or mouth breathers. The individuals were evaluated at two different periods, with a 2 year interval. At the first observation, the subjects were 11 years to 14 years 11 months of age and at the second observation, 13 years 4 months to 16 years 6 months of age. Height and thickness of the upper lip were measured on lateral cephalograms with the aid of a digital pachymeter. For each individual, electromyographic records were obtained of the orbicularis oris superior muscle at rest and in a series of 12 movements. The electromyographic data were normalized as a function of amplitude, for achievement of the percentage value of each movement. Pearson and Spearman correlation tests were applied.The results showed some correlation between morphology and muscle function (at a confidence level of 95 per cent). However, as the values of the correlation coefficient (r) were too low to establish associations between variables, it was concluded that the dimensions of the upper lip are not correlated with muscle activity.

Expression and function of fibroblast growth factor 10 and its receptor, fibroblast growth factor receptor 213, in bovine follicles

Some fibroblast growth factors (FGFs) affect ovarian follicle cell growth and/or differentiation. Whereas many FGFs activate several FGF receptors, FGF7 and FGF10 primarily activate only one, FGFR2B. As FGF7 is produced by bovine theca cells and acts on granulosa cells, we tested the hypothesis that FGF10 may also play a role in folliculogenesis in cattle. Reverse transcription-polymerase chain reaction demonstrated the presence of FGF10 mRNA in the oocytes and theca cells of the antral follicles, as well as in the preantral follicles. FGF10 protein was detected by immunohistochemistry in the oocytes of the preantral and antral follicles, and in the granulosa and theca cells of the antral follicles. FGF10 expression in theca cells changed during follicle development; mRNA abundance decreased with increasing follicular estradiol concentration in healthy follicles, and was lowest in highly atretic follicles. Culturing of granulosa cells in serum-free medium revealed FSH regulation of FGF10 receptor expression. The addition of FGF10 to cultured granulosa cells decreased the level of estradiol but did not alter cell proliferation. These data support a role for FGF10 in signaling to granulosa cells from theca cells and/or the oocyte.

Classical and molecular cytogenetic analysis in head and neck squamous cell carcinomas

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

5S rDNA chromosomal mapping and COI sequence analysis reveal differentiation among distinct populations of a characid fish Serrapinnus notomelas

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

Larval stages of Paradasygyius depressus (Bell, 1835) (Crustacea : Decapoda : Brachyura : Majidae) and a phylogenetic analysis for 21 genera of Majidae

All larval stages and the first crab instar of Paradasygyius depressus (Bell) were obtained in laboratory culture. Larval development consists of two zoeal stages, followed by the megalopa. Each larval stage is described in detail. Beginning with the first zoea, the duration of each stage was 4--7 (4.5 +/- 0.7), 4-5 (4.5 +/- 0.5), and 7 days, the megalopa and first crab instar appearing 11 +/- 1 and 15 days after hatching, respectively. A phylogenetic analysis of 21 genera of Majidae is provided based on 34 zoeal and three megalopal characters. The phylogenetic analysis resulted in four equally parsimonious trees 173 steps long (CI = 0.66, RI = 0.71, and RC = 0.47) supporting the monophyly of Oregoniinae, Majinae, and Inachinae (with the exclusion of Macrocheira de Haan incertae sedis). Based on general agreement of sister-group hypotheses, we provide sets of larval characters that define Oregoniinae, Majinae, and Inachinae. Our phylogenetic hypothesis suggests that Oregoniinae is the most basal clade within the Majidae, and Majinae and the clade (Epialtus H. Milne Edwards + Inachinae [excluding Macrocheira incertae sedis]) are sister taxa. Within Inachinae, all trees suggest that Inachus Weber and Macropodia Leach are sister taxa nested as the most derived clade, followed by Achaeus Leach, Pyromaia Stimpson, Paradasygyius Garth, Anasimus A. Milne-Edwards, and the most basal Stenorhynchus Lamarck. The sister-group relationships of the clade (Pisa Leach (Taliepus A. Milne-Edwards + Libinia Leach)), Mithrax Latreille and Microphrys H. Milne Edwards remained unresolved.

Structure and function of the foregut and salivary glands of the synanthropic diplopod Urostreptus atrobrunneus (Spirostreptidae)

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

Geographical variation in the golden-striped salamander, Chioglossa lusitanica Bocage, 1864 and the description of a newly recognized subspecies

Morphological and genetic data for the Iberian golden- striped salamander, Chioglossa lusitanica, demonstrate the existence of two groups with southern and northern ranges, connected by a zone of intergradation in central Portugal. Because reproductive isolation between them is incomplete we consider the groups to be subspecies. The type locality of C. lusitanica ( Bucaco near Lousa) is situated inside the mixed zone. This necessitates identification of the nominotypical subspecies. We sequenced a fragment of mitochondrial DNA from one of the species' syntypes and we determined what position over a latitudinal transect maximizes the morphological discrimination between the groups. Both approaches indicate that C. lusitanica from Bucaco represents the southern subspecies. A new subspecies of C. lusitanica is described from a northern locality ( Valongo near Porto in northwestern Portugal). A lectotype is designated for Chioglossa lusitanica.

Crystallization and preliminary crystallographic analysis of SMase I, a sphingomyelinase from Loxosceles laeta spider venom

SMase I, a 32 kDa sphingomyelinase found in Loxosceles laeta venom, is responsible for the major pathological effects of spider envenomation. This toxin has been cloned and functionally expressed as a fusion protein containing a 6 x His tag at its N-terminus to yield a 33 kDa protein [Fernandes-Pedrosa et al. (2002), Biochem. Biophys. Res. Commun. 298, 638 - 645]. The recombinant protein possesses all the biological properties ascribed to the whole L. laeta venom, including dermonecrotic and complement-dependent haemolytic activities. Dynamic light-scattering experiments conducted at 291 K demonstrate that the sample possesses a monomodal distribution, with a hydrodynamic radius of 3.57 nm. L. laeta SMase I was crystallized by the hanging-drop vapour-diffusion technique using the sparse-matrix method. Single crystals were obtained using a buffer solution consisting of 0.08 M HEPES and 0.9 M trisodium citrate, which was titrated to pH 7.5 using 0.25 M sodium hydroxide. Complete three-dimensional diffraction data were collected to 1.8 Angstrom at the Laboratorio Nacional de Luz Sincrotron (LNLS, Campinas, Brazil). The crystals belong to the hexagonal system ( space group P6(1) or P6(5)), with unit-cell parameters a = b = 140.6, c = 113.6 Angstrom. A search for heavy-atom derivatives has been initiated and elucidation of the crystal structure is currently in progress.

Transient and d.c. analysis of the operation mechanism of light-emitting electrochemical cells

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

Corrosion of the component phases presents in high copper dental amalgams. Application of electrochemical impedance spectroscopy and electrochemical noise analysis

The corrosion resistance of three of the constituent phases in high copper dental amalgams has been investigated by electrochemical methods in 0.9% NaCl solution. Polarization curves show corrosion potentials most positive for gamma(1)-Ag2Hg3, followed by Ag-Cu, and gamma-Ag3Sn in agreement with the order of corrosion resistance deduced from the corrosion currents. Complex plane impedance plots at the open circuit potential showed distorted semicircles with diffusional components at low frequency for Ag-Hg and Ag-Cu, while for gamma-Ag3Sn a layer of corrosion products is formed, partially or completely covering the surface of the electrode. Impedance and noise spectra have been compared in the frequency domain, and show good agreement. (C) 2004 Elsevier Ltd. All rights reserved.

Distribution transformers modeling with angular displacement: actual values and per unit analysis

In this work a detailed modeling of three-phase distribution transformers aimed at complementing well-known approaches is presented. Thus, incidence of angular displacement and tapping is taken into account in the proposed models, considering both actual values and per unit. The analysis is based on minimal data requirement: solely short-circuit admittance is needed since three-phase transformers are treated as non-magnetically-coupled single-phase transformers. In order to support the proposed methodology, results obtained through laboratory tests are presented.