Canine Subcutaneous Mast Cell Tumors: Cellular Proliferation and KIT Expression as Prognostic Indices

Molecular assays are widely used to prognosticate canine cutaneous mast cell tumors (MCT). There is limited information about these prognostic assays used on MCT that arise in the subcutis. The aims of this study were to evaluate the utility of KIT immunohistochemical labeling pattern, c-KIT mutational status (presence of internal tandem duplications in exon 11), and proliferation markers-including mitotic index, Ki67, and argyrophilic nucleolar organizing regions (AgNOR)-as independent prognostic markers for local recurrence and/or metastasis in canine subcutaneous MCT. A case-control design was used to analyze 60 subcutaneous MCT from 60 dogs, consisting of 24 dogs with subsequent local recurrence and 12 dogs with metastasis, as compared to dogs matched by breed, age, and sex with subcutaneous MCT that did not experience these events. Mitotic index, Ki67, the combination of Ki67 and AgNOR, and KIT cellular localization pattern were significantly associated with local recurrence and metastasis, thereby demonstrating their prognostic value for subcutaneous MCT. No internal tandem duplication mutations were detected in exon 11 of c-KIT in any tumors. Because c-KIT mutations have been demonstrated in only 20 to 30% of cutaneous MCT and primarily in tumors of higher grade, the number of subcutaneous MCT analyzed in this study may be insufficient to draw conclusions on the role c-KIT mutations in these tumors.

Evaluation of the apical infiltration after root canal disruption and obturation

The aim of this study was to evaluate two root canal filling techniques used in teeth that had their apical foramen disrupted and compare the apical infiltration with an ideal clinical situation. Twenty-seven freshly extracted single-rooted teeth were selected and radiographed to confirm the existence of a single and straight root canal. The crowns were removed at a mean distance of 11 mm from the apex. The teeth had the root canals instrumented and were randomly assigned to 3 groups (n=9): ND group-root canals were filled using the lateral compaction technique and no disruption was performed; DRF group-the apical constriction was disrupted by advancing a #40 K-file 1 mm beyond the original working length, the canals were reinstrumented to create an apical ledge at 1 mm from the apical foramen and were obturated with a master gutta-percha cone with same size as the last file used for reinstrumentation; DF group - the teeth had the apical constriction disrupted and the canals were obturated with a master gutta-percha cone that fit at 1 mm from the apex. The teeth were submitted to dye leakage test with Rhodamine B for 7 days, using vaccum on the initial 5 min. The teeth were sectioned longitudinally and the leakage was measured in a linear fashion from apex to crown. There was no statistically significant difference (p>0.05) between the groups that had the apical foramen disrupted (DF, DRF), but significant difference was found between the disrupted groups and the non-disrupted one (p<0.01). In conclusion, none of the evaluated techniques was able to prevent apical infiltration, so working length so the working length determination has to be established and maintained carefully.

Comparison between apo and complexed structures of bothropstoxin-I reveals the role of Lys122 and Ca2+-binding loop region for the catalytically inactive Lys49-PLA(2)s

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

Towards the prediction of essential genes by integration of network topology, cellular localization and biological process information

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

Prostate carcinogenesis induced by N-methyl-N-nitrosourea (mnu) in gerbils: Histopathological diagnosis and potential invasiveness mediated by extracellular matrix components

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

Nematode burdens and cellular responses in the abomasal mucosa and blood of Florida Native, Rambouillet and crossbreed lambs

This experiment was carried out to compare the worm burden and cellular responses in the abomasal mucosa and blood of Florida Native and Rambouillet lambs and also in animals produced by crosses of these two breeds (generations F1 and F2). Animals were exposed to infection by gastrointestinal nematodes on three different occasions. The first infection was natural, occurring while they were suckling lambs. After weaning, they were kept indoors for 53 days and then were allowed to graze a contaminated pasture for 50 days for a second natural infection. The third infection was an artificial challenge with 6000 Haemonchus contortus infective larvae. The highest mean fecal egg counts (FEC) values were found in Rambouillet lambs followed in decreasing order by F1, F2 and Florida Native lambs. Throughout the trial, most of the high mean packed cell volumes and plasma protein levels were recorded in the F2 lambs; in contrast, most of the low values were found in the Rambouillet lambs. During the natural infection period, the highest percentages of larvae in the fecal cultures of the lambs were Haemonchus. However, high percentages of Trichostrongylus larvae were found particularly in Florida Native lambs. The mean number of blood eosinophils increased after the artificial challenge, reached a peak 21 days after infection and then declined. The highest and lowest blood eosinophil means were recorded in F2 and Florida Native lambs, respectively. The H. contortus burden was significantly higher in Rambouillet and in F1 lambs than in Florida Native and F2 lambs (p < 0.05), while no significant differences were recorded among eosinophil, mast cell and globule leucocyte counts in the abomasal mucosa (p > 0.05). The highest correlation coefficient recorded at the end of this study was between FEC and worm burden (r = 0.7). These two parameters showed a moderate negative correlation with PCV, plasma protein and mast cell counts in the abomasal mucosa. The results obtained in this study indicate that crossbreeding Florida Native and Rambouillet sheep can be a rapid way to combine and improve the characteristics of these two breeds. The parasitological results were promising. however, more studies will be necessary to verify the impact of crossbreeding in other traits. (C) 1999 Elsevier B.V. B.V. All rights reserved.

Mastoparan effects in skeletal muscle damage: An ultrastructural view until now concealed

Animal venoms have been valuable sources for development of new drugs and important tools to understand cellular functioning in health and disease. The venom of Polybia paulista, a neotropical social wasp belonging to the subfamily Polistinae, has been sampled by headspace solid phase microextraction and analyzed by gas chromatography-mass spectrometry. Recent study has shown that mastoparan, a major basic peptide isolated from the venom, reproduces the myotoxic effect of the whole venom. In this study, Polybia-MPII mastoparan was synthesized and studies using transmission electron microscopy were carried out in mice tibial anterior muscle to identify the subcellular targets of its myotoxic action. The effects were followed at 3 and 24 h, 3, 7, and 21 days after mastoparan (0.25 mu g/mu L) intramuscular injection. The peptide caused disruption of the sarcolemma and collapse of myofibril arrangement in myofibers. As a consequence, fibers presented heteromorphic amorphous masses of agglutinated myofilaments very often intermingled with denuded sarcoplasmic areas sometimes only surrounded by a persistent basal lamina. To a lesser extent, a number of fibers apparently did not present sarcolemma rupture but instead appeared with multiple small vacuoles. The results showed that sarcolemma, sarcoplasmic reticulum (SR), and mitochondria were the main targets for mastoparan. In addition, a number of fibers showed apoptotic-like nuclei suggesting that the peptide causes death both by necrosis and apoptosis. This study presents a hitherto unexplored view of the effects of mastoparan in skeletal muscle and contributes to discuss how the known pharmacology of the peptide is reflected in the sarcolemma, SR, mitochondria, and nucleus of muscle fibers, apparently its subcellular targets.

Permethrin-induced ultrastructural changes in oocytes of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) semi-engorged females

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

Cellular and extracellular behavior in the gerbil (Meriones unguiculatus) ventral prostate following different types of castration and the consequences of testosterone replacement

Mongolian gerbils (Meriones unguiculatus) were grouped into two experimental groups: GEx.01 suffered orchiectomy and after 30 days received doses of testosterone cipionate (T), while GEx.02 received weekly and alternated doses of the anti-androgens cyproterone acetate and flutamide for 30 days, and the animals were then euthanized. Structural evaluation reveals a more intense reduction in epithelial height in GEx.02. Smooth muscle cells (SMC) presented a star-shaped aspect after 30 days of hormonal ablation and basal membrane was shown to be more intensely grooved in GEx.01. In both groups, after hormonal replacement, recovery in epithelial height could be noted and the SMC presented its phenotypes, but an increase in RER was seen, characterizing a modulation from its contractile to secreting phenotype. In conclusion, the prostate presented involution capacity after androgen ablation and the ability to reorganize after hormonal replacement, but events resulting from orchiectomy and subsequent T replacement were shown to be more aggressive to the prostate. (c) 2006 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.

Cellular changes in the prostatic stroma of glucocorticoid-treated rats

Glucocorticoid hormones (GCs) have been widely used for the treatment of prostate cancer because of their inhibitory property against tumour growth. However, their mechanism of action in the prostate has received little attention. Excess GCs can lead to peripheral insulin resistance resulting in hyperglycaemia and hyperinsulinaemia. Insulin plays an important role as a cellular stimulant and high levels are related to low levels of androgens. Our objective has been to describe the effects of insulin resistance induced by dexamethasone treatment on the morphology of rat ventral prostate. Mate adult Wistar rats received daily intraperitoneal injections of dexamethasone or saline for five consecutive days after which the rats were killed and the ventral prostate was removed, weighed and prepared for conventional and transmission electron microscopy (TEM). Dexamethasone treatment resulted in atrophy and decreased proliferative activity of prostatic epithelial cells. TEM analysis revealed changes in the epithelium-stroma interface, with some interruptions in the basement membrane. Fibroblasts showed a secretory phenotype with dilated endoplasmic reticulum. Smooth muscle cells exhibited a contractile pattern with 50% atrophy, an irregular membrane and twisted nuclei. Mitochondrial alterations, such as enlarged size and high electron density in the mitochondrial matrix, were also detected in smooth muscle cells. Insulin resistance induced by dexamethasone is thus associated with epithelial atrophy similar to that described for diabetic rats. However, GCs are responsible for morphological changes in the stromal cell population suggesting the activation of fibroblasts and atrophy of the smooth muscle cells.

Long-term inhibition of 5-alpha reductase and aromatase changes the cellular and extracellular compartments in gerbil ventral prostate at different postnatal ages

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

Maximum entropy, fractal dimension and lacunarity in quantification of cellular rejection in myocardial biopsy of patients submitted to heart transplantation

This paper presents a method for the quantification of cellular rejection in endomyocardial biopsies of patients submitted to heart transplant. The model is based on automatic multilevel thresholding, which employs histogram quantification techniques, histogram slope percentage analysis and the calculation of maximum entropy. The structures were quantified with the aid of the multi-scale fractal dimension and lacunarity for the identification of behavior patterns in myocardial cellular rejection in order to determine the most adequate treatment for each case.

Demonstration of antibody and cellular immune response to brain extract in West and Lennox-Gastaut syndromes

Investigamos a resposta imunológica celular e humoral frente a extrato salino de tecido cerebral em 9 pacientes com síndrome de Lennox-Gastaut, 15 pacientes com síndrome de West e 20 crianças normais. A técnica de imunodifusão dupla em gel de agar (Ouchterlony) evidenciou em todos os pacientes, altos níveis de um anticorpo precipitante contra o extrato salino de tecido cerebral. O teste de inibição de migração de leucócitos com o mesmo antígeno mostrou-se positivo na maioria dos pacientes. O possível papel destas respostas autoimmunes na patogenia das sindromes de West e Lennox-Gastaut é discutido.

Histological characterization of cellular types during Scinax fuscovarius oogenesis (Lutz) (Anura, Hylidae)

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