Estudo in vitro dos efeitos da BMP-2 e do seu antagonista Noggin sobre a proliferação e migração celulares em carcinoma epidermóide de língua

Oral squamous cell carcinoma (OSCC) is the most prevalent malignancy in the oral cavity and reach a large number of individuals, has become an important public health problem. Studies have demonstrated changes in pathway components BMP in various types of cancers as prostate, colon, breast, gastric and OSCCs. Is the current knowledge that these proteins may exert pro-tumor effect in more advanced stages of neoplastic development coming to favor progression and invasion tumor. The inhibition of the signaling pathway BMP-2 through its antagonists, have shown positive results of antitumor activity and use of Noggin may be a novel therapeutic target for cancer. Given this evidence and the few studies with BMP-2, Noggin and OSCC, the objective of this research was to evaluate the effect of BMP-2 and its antagonist Noggin on proliferation and migration cell in line of cell cultures of human tongue squamous cell carcinoma (SCC25). The study was divided in three groups, a control group, where SCC25 cells suffered no treatment, a BMP-2 group, in which cells were treated with 100ng/ml of BMP-2 and a group of cells that were treated with 100ng/ml of Noggin. For the proliferation assay and cell cycle were established three time intervals (24, 48 and 72 hours). Proliferative activity was investigated by trypan blue and cell cycle analysis by staining with propidium iodide flow cytometry. The potential for migration / invasion of SCC25 cells was performing by a cell invasion assay using Matrigel in a 48-hour interval. The proliferation curve showed a higher proliferation in cells treated with BMP-2 in 72 hours (p < 0.05), and lower overgrowth and cell viability in Noggin group. Recombinant proteins favored a greater percentage of cells in cell cycle phase Go/G1 with a statistically significant difference in the interval of 24 hours (p < 0.05). BMP- 2 produced a greater invasion of cells studied as well as its antagonist Noggin inhibits invasion of cells (p < 0.05). Thus, these results indicate that BMP-2 promotes malignant phenotype, dues stimulates proliferation and invasion of SCC25 cells and, its antagonist Noggin may be an alternative treatment, due to inhibit the tumor progression

Mineral trioxide aggregate stimulates macrophages and mast cells to release neutrophil chemotactic factors: role of IL-1 beta, MIP-2 and LTB4

Objective. In the present study, the role of macrophages and mast cells in mineral trioxide aggregate (MTA)-induced release of neutrophil chemotactic factor was investigated.Study design. MTA suspension (50 mg/mL) was plated over inserts on macrophages or mast cells for 90 minutes. Untreated cells served as controls. Cells were washed and cultured for 90 minutes in RPMI without the stimuli. Macrophages and mast cell supernatants were injected intraperitoneally (0.5 mL/cavity), and neutrophil migration was assessed 6 hours later. In some experiments, cells were incubated for 30 minutes with dexamethasone (DEX, 10 mu M/well), BWA4C (BW, 100 mu M/well) or U75302 (U75, 10 mu M/well). The concentration of Leukotriene B-4 (LTB4) in the cell-free supernatant from mast cells and macrophage culture was measured by ELISA.Results. Supernatants from MTA-stimulated macrophages and mast cells caused neutrophil migration. The release of neutrophil chemotactic factor by macrophages and mast cells was significantly inhibited by DEX, BW, or U75. Macrophages and mast cells expressed mRNA for interleukin-1 (IL-1)beta and macrophage inflammatory protein-2 (MIP-2) and the pretreatment of macrophages and mast cells with DEX, BW, or U75 significantly altered IL-1 beta and MIP-2 mRNA expression. LTB4 was detected in the MTA-stimulated macrophage supernatant but not mast cells.Conclusions. MTA-induces the release of neutrophil chemotactic factor substances from macrophages and mast cells with participation of IL-1 beta, MIP-2, and LTB4. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010; 109: e135-e142)

Mechanism of calcium hydroxide-induced neutrophil migration into air-pouch cavity

The aim of this study was to investigate cellular migration induced by calcium hydroxide to air-pouch cavities in mice. The migration was more specific to neutrophil and was dose and time dependent (peaking 96 h after stimulation). This migration was inhibited by pretreatment with thalidomide, indomethacin, MK886, meloxicam, dexamethasone, MK886 associated with indomethacin, and MK886 associated with indomethacin and dexamethasone. The air-pouch exudate from animals stimulated with calcium hydroxide showed an increase of leukotriene-B4 (LTB4), interleukin-1 beta, tumor necrosis factor alpha (TNF-alpha), cytokine-induced neutrophil chemoattractant (KC), and macrophage inflammatory protein 2 (MIP-2) release. Pretreatment with 3% thioglycollate increased the macrophage population in the air pouch but did not change neutrophil migration. Depleting the resident mast cells through chronic pretreatment with compound 48/80 did not alter neutrophil migration in response to calcium hydroxide. It was possible to conclude that calcium hydroxide-induced neutrophil migration to the air-pouch cavity in mice is mediated by LTB4, TNF-alpha, KC, MIP-2, and prostaglandins, but it was not dependent on macrophages or mast cells.

In vitro comparison of 1.5 mm vs. 2.0 mm screws for fixation in the sagittal split osteotomy

Purpose: Numerous "in vitro" investigations have been conducted to evaluate the role of screw size and pattern in determining optimal resistance to deformation, often these have been controversial. The aim of this study was to evaluate the effect of screw size and insertion technique on the stability of sagittal split osteotomies.Materials and methods: This study used twenty polyurethane replicas of human hemimandibles with a prefabricated sagittal split ramus osteotomy (SSRO). The hemimandibles were stabilized with 1.5 mm and 2.0 mm titanium screws inserted in an inverted L configuration. All specimens were tested to determine the strength and stability of the fixation.Results: In all cases there was failure of the synthetic bone before there was any evidence of screw failure. There were no significant differences in the load necessary to make the construct fail between the 1.5 or 2.0 mm screw sizes.Conclusion: There was no statistically significant difference between the strengths achieved with screws of 1.5 and 2.0 mm diameters for fixation of SSRO performed in synthetic mandibles. There was no fracture of the 1.5 mm or 2.0 mm diameter screws in any of the tests. 1.5 mm diameter screws in an inverted L pattern have as much stability and mechanical resistance as a 2.0 mm screw, may be safely used for this procedure. (C) 2010 European Association for Cranio-Maxillo-Facial Surgery.

HTRIdb: an open-access database for experimentally verified human transcriptional regulation interactions

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

Knockdown resistance in pyrethroid-resistant horn fly (Diptera: Muscidae) populations in Brazil

Com o objetivo de verificar a ocorrência e determinar a frequência da mutação kdr (knock down resistance) em populações de Haematobia irritans (mosca-dos-chifres) resistentes aos piretróides, foram analisados 1.804 indivíduos de 37 populações de todas as Regiões do Brasil. Com exceção da Região Nordeste, o kdr (knock down resistance gene) foi encontrado em populações de todas as regiões. A mutação não foi detectada em 87,08% dos indivíduos. Entretanto, o gene foi amplificado de 12,92% das moscas, das quais 11,70% se mostraram heterozigotas resistentes e 1,22% homozigotas resistentes. em todas as populações verificou-se equilíbrio de acordo com a Lei de Hardy e Weinberg, exceto uma com excesso de heterozigotos. Entretanto, quando agrupamos diferentes populações numa metapopulação de acordo com a região geográfica, é possível observar um desvio nas populações Centro-Oeste, Sul e Sudeste, indicando isolamento populacional e que a ocorrência do kdr é provavelmente um efeito independente, talvez refletindo a estratégia de uso do inseticida de cada produtor. Apesar da resistência aos piretróides estar disseminada por todo o país, apenas 48% das populações resistentes apresentaram o kdr, e a frequência de indivíduos kdr nas populações resistentes se mostrou bastante baixa. À exceção da Região Nordeste, o mecanismo de resistência ligado ao kdr ocorre em todo o país.

Acute exercise reverses aged-induced impairments in insulin signaling in rodent skeletal muscle

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

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)

Reduction of the nutritional values of diets for hens through supplementation with phytase

Objetivou-se avaliar os efeitos da redução dos níveis nutricionais de dietas para poedeiras marrons em pico de postura sobre o desempenho das aves, a qualidade dos ovos e os parâmetros ósseos. No total foram avaliadas oito dietas, cada uma com cinco repetições de oito aves: 1. controle: formulada para suprir as exigências das aves; 2) controle + 0,03% ou 600 FTU; 3) 15% PB; 2.800 kcal de EM; 3,8% Ca e 0,28% Pd; 4) dieta 3 + 0,03% ou 600 FTU; 5) 14% PB; 2.750 kcal de EM; 3,4% Ca e 0,23% Pd; 6) dieta 5 + 0,03% ou 600 FTU; 7) 13% PB; 2.700 kcal de EM; 3,0% Ca e 0,18% Pd; e 8) dieta 7 + 0,03% ou 600 FTU. A redução dos níveis nutricionais prejudicou o desempenho das aves, principalmente a produção e massa de ovos, que melhorou com a inclusão de fitase nas dietas. Os parâmetros de qualidade dos ovos, no entanto, não se alteraram com a redução até o nível de 15% de PB; 2.800 kcal de EM/kg; 3,80% Ca e 0,280% Pd, o que não se repetiu nos níveis com menor densidade nutricional. Não houve efeito da redução dos níveis nutricionais sobre os parâmetros ósseos avaliados, exceto a resistência, que melhorou com a suplementação de fitase no nível de 600 FTU. A redução nos níveis nutricionais da dieta para 15% de PB, 2.800 kcal/kg de EM, 3,8% de Ca e 0,280% de Pd com a adição de 600 FTU não prejudica a produção nem a massa de ovos e melhora a saúde óssea das aves.

INGAP-PP up-regulates the expression of genes and proteins related to K-ATP(+) channels and ameliorates Ca2+ handling in cultured adult rat islets

Islet Neogenesis Associated Protein (INGAP) increases pancreatic beta-cell mass and potentiates glucose-induced insulin secretion. Here, we investigated the effects of the pentadecapeptide INGAP-PP in adult cultured rat islets upon the expression of proteins constitutive of the K-ATP(+) channel, Ca2+ handling, and insulin secretion. The islets were cultured in RPMI medium with or without INGAP-PP for four days. Thereafter, gene (RT-PCR) and protein expression (Western blotting) of Foxa2, SUR1 and Kir6.2, cytoplasmic Ca2+ ([Ca2+](i)), static and dynamic insulin secretion, and Rb-86 efflux were measured. INGAP-PP increased the expression levels of Kir6.2, SUR1 and Foxa2 genes, and SUR1 and Foxa2 proteins. INGAP-PP cultured islets released significantly more insulin in response to 40 mM KCl and 100 mu M tolbutamide. INGAP-PP shifted to the left the dose-response curve of insulin secretion to increasing concentrations of glucose (EC50 of 10.0 +/- 0.4 vs. 13.7 +/- 1.5 mM glucose of the controls). It also increased the first phase of insulin secretion elicited by either 22.2 mM glucose or 100 mu M tolbutamide and accelerated the velocity of glucose-induced reduction of Rb-86 efflux in perifused islets. These effects were accompanied by a significant increase in [Ca2+](i) and the maintenance of a considerable degree of [Ca2+](i) oscillations. These results confirm that the enhancing effect of INGAP-PP upon insulin release, elicited by different secretagogues, is due to an improvement of the secretory function in cultured islets. Such improvement is due, at least partly, to an increased K-ATP(+) channel protein expression and/or changing in the kinetic properties of these channels and augmented [Ca2+](i) response. Accordingly, INGAP-PP could potentially be used to maintain the functional integrity of cultured islets and eventually, for the prevention and treatment of diabetes. (C) 2008 Elsevier B.V. All rights reserved.

Two-matrix string model as constrained (2+1)-dimensional integrable system

We show that the 2-matrix string model corresponds to a coupled system of 2 + 1-dimensional KP and modified KP ((m)KP2+1) integrable equations subject to a specific symmetry constraint. The latter together with the Miura-Konopelchenko map for (m)KP2+1 are the continuum incarnation of the matrix string equation. The (m)KP2+1 Miura and Backhand transformations are natural consequences of the underlying lattice structure. The constrained (m)KP2+1 system is equivalent to a 1 + 1-dimensional generalized KP-KdV hierarchy related to graded SL(3,1). We provide an explicit representation of this hierarchy, including the associated W(2,1)-algebra of the second Hamiltonian structure, in terms of free currents.

MTA-induced neutrophil recruitment: a mechanism dependent on IL-1β, MIP-2, and LTB4

Objective: The objective of this study was to investigate the mediators and the resident peritoneal cells involved in the neutrophil migration (NM) induced by mineral trioxide aggregate (MTA) in mice. Study design: MTA (25 mg/cavity) was injected into normal and pretreated peritoneal cavities (PC) with indomethacin (IND), dexamethasone (DEX), BWA4C, U75302, antimacrophage inflammatory protein-2 (MIP-2), and anti-interleukin-1β (IL-1β) antibodies and the NM was determined. The role of macrophage (MO) and mast cells (MAST) was determined by administration of thioglycollate 3% or 48/80 compound, respectively. The concentration of IL-1β and MIP-2 exudates was measured by ELISA. Results: MTA induced dose- and time-dependent NM into mice PC, with the participation of MO and MAST. NM was inhibited by DEX, BWA4C, and U75302, as well as anti-MIP-2 and anti-IL-1β antibodies. In the exudates, IL-1β and MIP-2 were detected. Conclusions: This study suggests that MTA induces NM via a mechanism dependent on MAST and MO mediated by IL-1β, MIP-2, and LTB4. © 2008 Mosby, Inc. All rights reserved.

Perfil longitudinal e a aplicação do índice de gradiente (RDE) no rio dos Patos, bacia hidrográfica do rio Ivaí, PR

This paper presents the results of the longitudinal profile analysis and the application of gradient index (RDE) in the Patos River, a tributary of the Ivaí River basin, PR. The goal is identify and quantify the anomalous zones along river course concerning their genesis - activity neotectonic or/and lithological imposition. The Patos River has a length of 127 km and a relief amplitude of 660 m, The occurrence of an important knickpoint in the middle course the Patos River can be divided into two stretches. Of the 22 sections measured, 10 were considered anomalous, distributed in both upper and lower course. In the upper course, over a basaltic bedrock of the Serra Geral Formation the RDE values are lower than those in the lower course, where river flows over sedimentary rocks of the Terezina Formation. The 2 nd order anomalies are related to lithological changes, relief alignments and channel confluences - as in the case of São João River. The 1st order anomalies are linked to differences in rock resistance associated with structural control, which provides the formation of waterfalls.

Association of IGF1 and KDM5A polymorphisms with performance, fatness and carcass traits in chickens

Two functional and positional candidate genes were selected in a region of chicken chromosome 1 (GGA1), based on their biological roles, and also where several quantitative trait loci (QTL) have been mapped and associated with performance, fatness and carcass traits in chickens. The insulin-like growth factor 1 (IGF1) gene has been associated with several physiological functions related to growth. The lysine (K)-specific demethylase 5A (KDM5A) gene participates in the epigenetic regulation of genes involved with the cell cycle. Our objective was to find associations of selected single-nucleotide polymorphisms (SNPs) in these genes with performance, fatness and carcass traits in 165 F2 chickens from a resource population. In the IGF1 gene, 17 SNPs were detected, and in the KDM5A gene, nine SNPs were detected. IGF1 SNP c. 47673G > A was associated with body weight and haematocrit percentage, and also with feed intake and percentages of abdominal fat and gizzard genotype × sex interactions. KDM5A SNP c. 34208C > T genotype × sex interaction affected body weight, feed intake, percentages of abdominal fat (p = 0. 0001), carcass, gizzard and haematocrit. A strong association of the diplotype × sex interaction (p < 0. 0001) with abdominal fat was observed, and also associations with body weight, feed intake, percentages of carcass, drums and thighs, gizzard and haematocrit. Our findings suggest that the KDM5A gene might play an important role in the abdominal fat deposition in chickens. The IGF1 and KDM5A genes are strong candidates to explain the QTL mapped in this region of GGA1. © 2012 Institute of Plant Genetics, Polish Academy of Sciences, Poznan.

Hormonal treatment of dairy goats affected by hydrometra associated or not with ovarian follicular cyst

Hydrometra is considered a very important pathological condition, because it represents one of the main causes of temporary infertility in dairy goats. The objective was (i) to evaluate a protocol for the treatment of hydrometra associated (n=2) or not (n=17) with follicular ovarian cyst in 19 dairy goats and (ii) to assess its reproductive efficiency after treatment. For this purpose, 10. mg of dinoprost (PGF) divided in two equal doses were administered to all animals intravulvosubmucosally on Days 0 and 10. In addition, 500. IU hCG were administered on Day 7. Ultrasound exams were performed in all females from Days 0 to 3, 7 and 10 to 13 of treatment, in order to evaluate uterus drainage after each treatment. Goats were monitored for estrus after both treatments and mated after the second dose of PGF. Blood samples were collected from 11 goats to determine plasma progesterone concentrations before, during and after treatment. Of the 19 goats treated, 16 lost weight after the first dose, probably due to uterine discharge. Complete drainage of uterine fluid was observed in 11/19 (57.9%) and 17/19 (89.5%) after the first and second doses, respectively. Afterwards, we diagnosed 2 more goats with follicular cysts, for a total of 21.1% (4/19) of animals exhibiting hydrometra and ovarian cyst concomitantly. In one doe the diagnosis was on Day 2 and in the other on Day 11. All does showed progesterone concentrations superior to 1. ng/mL at Day 0, with an average of 10.6 ± 1.4. ng/mL. Out of the 10 goats mated, only two became pregnant after treatment, corresponding to 10.5% of the total (2/19). Although prostaglandin was effective to drain the uterine fluid and led to the onset of estrus, it did not improve the pregnancy rate. The use of hCG in female goats was not effective in luteinizing the cysts. It can be concluded that hydrometra alone or associated with ovarian follicular cyst may adversely affect goat reproductive performance. © 2012 Elsevier B.V.