AKT and AMPK Activation after High-Fat and High-Glucose In Vitro Treatment of Prostate Epithelial Cells

Considering the increasing consumption of saturated fat and glucose in diets worldwide and its possible association to carcinogenesis, this investigation analysed the proliferation profile of nonmalignant human prostate epithelial cells after exposure to elevated levels of fat and glucose. PNT1A cells were cultured with palmitate (100 or 200 mu M) and/or glucose (450mg/dl) for 24 or 48 h. Treated cells were evaluated for viability test and cell proliferation (MTS assay). AKT and AMPK phosphorylation status were analysed by Western blotting. After 24 h of high-fat alone or associated with high-glucose treatment, there was an increase in AMPK and AKT activation associated to unchanged MTS-cell proliferation. Following 48 h of high-fat but not high-glucose alone, cells decreased AMPK activation and maintained elevated AKT levels. These data were associated to increased cell proliferation after further high-fat treatment. After longer high-fat exposure, MTS revealed that cells remained proliferating. High-glucose alone or associated to high-fat treatment was not able to increase cell proliferation and AKT activation. A high-fat medium containing 100 mu M of palmitate stimulates proliferation in PNT1A cells by decreasing the activation of AMPK and increasing activation of AKT after longer exposure time. These findings improve the knowledge about the negative effect of high levels of this saturated fatty acid on proliferative disorders of prostate.

Stabilization of dentin matrix after cross-linking treatments, in vitro

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Dioctadecyldimethylammonium:Monoolein Nanocarriers for Efficient in Vitro Gene Silencing

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

Modelos in vitro e in vivo aplicados à avaliação de toxicidade e eficácia de fitocosmético: Serjania marginata Casar. (Sapindaceae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo comparativo in vitro da resistência ao cisalhamento de sistemas adesivos em dentina humana: uso de hipoclorito de sódio em sistemas convencionais ou primers acidificados

Pós-graduação em Odontologia Restauradora - ICT

In Vitro Assessment of Mutagenic And Genotoxic Effects of Coumarin Derivatives 6,7-Dihydroxycoumarin and 4-Methylesculetin

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

Cultivo celular in vitro: importância para a pesquisa biomédica e dimensão da problemática de autenticação de linhagens celulares

Experimental models composed by human and animal cell lines are simplified and informative, allowing them to be widely used for biomedical research. Most laboratories that use in vitro cultivated cells maintain a variation of cell lines stored and cultivated. Therefore, misidentification and cross-contamination events can happen during cell lines handling. This problem can generate a repertoire of dubious results and papers, which may prejudice biomedical research. Recently it was created the International Cell Line Authentication Committee (ICLAC), which aims to spread knowledge about cross-contamination and misidentification of in vitro cell lines. Despite of the efforts spent trying to aware scientific community about the importance of the correct identification of cells, the number of papers based on misidentified cell lines it´s still worrying, compromising the reliability of out coming results and conclusions regarding them. The present study aims to analyze and discuss the main advantages and limitations of eukaryote in vitro cell lines use, characterizing the cell lines authentication problems. Therefore, compilation and critical analyses of literature data was realized, aiming to improve the understanding about this subject. Based on information about 445 cell lines with issues published by ICLAC it´s clear that contamination in human cell lines represented 89,2 % of mentioned problems. HeLa cell line was the responsible for most contamination, especially in 92 normal tissue cell lines, representing 44,6% of the contamination. These results reinforce the importance of periodic maintenance of cell lines cultures by labs and implementation of authentication methods as polymorphic STRs, besides obtaining cell lines from reliable sources and cell banks

Avaliação do Efeito Fotodinâmico sobre o Crescimento in vitro de Pythium insidiosum

Photodynamic Therapy (PDT) is a therapeutic method which employs a photosensitizer and light to cause cellular death. The chemical compounds have low or none toxicity for hosts cells. Under the incidence of light, in an appropriated wavelength, these chemical compounds produce reactive oxygen which affects the biomolecules of the target-cells. The specific illumination of the affected area increases the selectivity of the therapy, since the photodynamic process occurs only in the irradiated area. Pythiosis, for instance, is a life-threatening emerging disease caused by a fungus-like organism called Pythium insidiosum. The disease occurs in man and other animals, being mostly observed in horses. Human pythiosis may present as ophthalmic, cutaneous-subcutaneous and systemic forms of lesions. Due to the fact that P. insidiosum is not a true fungus, it is refractory to most antifungal drugs and the treatment of the disease is difficult. Extensive surgery procedures, such as limb amputation, are the treatment of choice, however relapses may occur frequently. Although not totally effective, the use of immunotherapy associated to surgery have shown some results. Considering that pythiosis is an emerging disease few explored in its etiological and therapeutic aspects, which are limited and few effective, it is of great importance to encourage the development of researches for new strategies of treatment. In this sense, it was evaluated the effect of PDT on in vitro growth of the pathogen employing two chemical compounds as photosensitizer, porphyrin and chlorine, at different concentrations in combination with several energetic dosages. Porphyrin showed inhibition of growth at 25mg/mL with 100J/cm2 of energetic dosage and chlorine showed similar results employing low concentrations (0,7, 1,0 and 1,3mg/mL) with 70J/cm2 of energetic dosage... (Complete abstract click electronic access below)

Caracterização in vitro de técnica biomagnética empregando magnetômetros 3D para avaliar tempo de trânsito esofagiano

The biomagnetic techniques use different magnetic field detectors to measure parameters of the human physiology. Those techniques present the advantage of being noninvasive and radiation free. Among them we can show up the Superconducting Quantum Interference Device (SQUID), the Current Alternate Biosusceptometry (ACB) and, more recently, the employment of anisotropic magnetoresistive sensors. Those magnetic sensors have a low cost and good sensitivity to measure different physiological parameters using magnetic markers. The biomagnetic techniques have being used successfully through study on the characteristics of the gastrointestinal tract. Recent research, the magnetoresistors were used to evaluate the transit time and localization of magnetic sources in different parts of the gastrointestinal tract. The objective of this work is the characterization, with in vitro tests, of a biomagnetic instrumentation using two 3-axis magnetoresistors arranged in a gradiometric coplanar setup to evaluate esophageal transit time, analyze and compare the results of experimental signals and the magnetic theory, as well as evaluate the instrumentation gain with use of tri-axial sensor front to the mono-axial sensor. The instrumentation is composed by two three-axis sensing magnetometers, precision power supply and amplifier electronic circuits. The sensors fixed in a coplanar setup were separate by distance of 18 cm. The sensitivity tests had been carried through using a cylindrical magnet (ø = 4 mm and h = 4 mm) of neodymium-iron-boron (grid 35). The tests were done moving the permanent magnet on the sensors parallel axis, simulating the food transit in... (Complete abstract click electronic access below)

Avaliação in vitro da ação de substâncias químicas auxiliares, clorexidina e medicações intracanais sobre C. albicans, E. faecalis, E. coli e sua endotoxina em canais radiculares: [recurso eletrônico]

The purpose of this study was to evaluate the antimicrobial activity of chlorhexidine gel 2% as auxiliary chemical substance on the biomechanical preparation (BMP) and medication intracanal (ICM) on C. albicans, E. faecalis, E. coli and their endotoxin in root canals. We used 48 single-rooted human teeth divided into four groups according to dressing ICM: 1) Ca(OH)2 + pyrogen-free saline solution; 2) 2% chlorhexidine gel (CLX); 3) Ca(OH)2 + CLX, and; 4) pyrogen-free saline solution (control group). Were collected the contents of root canals to confirm the presence of microorganisms (confirmation), immediately after instrumentation (1st collection), after 7 days of the BMP (2nd collection), after 14 days of the action of ICM (3rd Collection) and 7 days after removal of the ICM (4 th collection). Were performed: the evaluation of antimicrobial activity and the content analysis of endotoxins for all sampling tests. The results were statistically analyzed using Kruskall-Wallis and Dunn tests with a significance of 5%. It was found that the CLX as auxiliary chemical substance has significantly reduced microorganisms confirmation collection when compared. In relation to the neutralization of endotoxin, it was found that the 1st and 2nd collections presented a decrease of 92.03% and 98.10% in mean percentage respectively, when compared to the confirmation collection. In the 3rd and 4th samplings, the Ca (OH)2 + CLX group showed the best results. It was concluded that the BMP and the ICM were able to eliminate the tested microrganisms, however, they were not able to completely eliminate endotoxins root canal

Antitumor Activity of Kielmeyera Coriacea Leaf Constituents in Experimental Melanoma, Tested in Vitro and in Vivo in Syngeneic Mice

Purpose: The antitumor activity of Kielmeyera coriacea (Clusiaceae), a medicinal plant used in the treatment of parasitic, as well as fungal and bacterial infections by the Brazilian Cerrado population, was investigated. Methods: A chloroform extract (CE) of K. coriacea was tested in the murine melanoma cell line (B16F10-Nex2) and a panel of human tumor cell lines. Tumor cell migration was determined by the wound-healing assay and the in vivo antitumor activity of CE was investigated in a melanoma cell metastatic model. 1H NMR and GC/MS were used to determine CE chemical composition. Results: We found that CE exhibited strong cytotoxic activity against murine melanoma cells and a panel of human tumor cell lines in vitro. CE also inhibited growth of B16F10- Nex2 cells at sub lethal concentrations, inducing cell cycle arrest at S phase, and inhibition of tumor cell migration. Most importantly, administration of CE significantly reduced the number of melanoma metastatic nodules in vivo. Chemical analysis of CE indicated the presence of the long chain fatty compounds, 1-eicosanol, 1-docosanol, and 2-nonadecanone as main constituents. Conclusion: These results indicate that K. coriacea is a promising medicinal plant in cancer therapy exhibiting antitumor activity both in vitro and in vivo against different tumor cell lines.

Resveratrol como meio de estocagem para dentes avulsionados: estudo in vitro da viabilidade celular

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação dos efeitos de aloe-emodin em células uroteliais imortalizadas e neoplásicas cultivadas in vitro

Aloe vera (Aloe barbadensis Miller), popularly known in Brazil as babosa, has a long history of use as medicinal plant for different therapeutic purposes. The components of the plant extract are present in various products of human use, mainly for nutritional and cosmetics purposes. However, some studies suggest that this extract might also have carcinogenic activity. The aloe vera extract is a complex mixture of bioactive compounds. The study of isolated compounds may contribute to elucidate contradictory results about the effects related to the consumption of the plant, as well as their mechanisms of action. One of the most important compound from Aloe vera is aloe-emodin, which is a secondary metabolite generated in the intestinal tract. Putative antimicrobial and antitumor effects were previously attributed to aloe-emodin. Although the exposure of urothelial cells to aloe-emodin was already reported in the literature, only one study showed its effects on urothelial cells, suggesting that aloe-emodin inhibits the viability of T24 cancer cells due to apoptosis induction. Since there is no sufficient information about the effects of aloe-emodin on urothelial cells, and low efficiency in the treatment of bladder cancer currently, the present study aims to evaluate the hypothesis that the treatment with aloe-emodin could impact the behavior of other urothelial cell lines in vitro. Therefore, the in vitro IC50 exposure of aloe-emodin to human immortalized neoplastic urothelial cells will be determinated in order to verify possible differences in the behavior of urothelial cells in vitro treated with aloe-emodin in comparison with untreated cells. Furthermore, differences between cell lines will be also evaluated

In vitro biomechanical comparison of six different fixation methods following 5-mm sagittal split advancement osteotomies

The sagittal split ramus osteotomy (SSRO) is a surgical technique used widely to treat many congenital and acquired mandibular discrepancies. Stabilization of the osteotomy site and the potential for skeletal relapse after the procedure are still major problems. The aim of this study was to compare the mechanical stability of six methods of rigid fixation in SSRO using a biomechanical test model. Sixty polyurethane replicas of human hemimandibles were divided into six groups. In group I, the osteotomies were fixed with two four-hole titanium miniplates; in group II, with one four-hole miniplate; in group III, with one four-hole miniplate + a bicortical screw; in group IV, with a grid miniplate; in group V, with a four-hole locking miniplate; and in group VI, with a six-hole miniplate. A linear load in the premolar region was applied to the hemimandibles. The resistance forces (N) needed to displace the distal segment by 1, 3, and 5 mm were recorded and the data transmitted from the load cell to a computer. One-way analysis of variance with Tukey's post hoc test was performed to compare the means between groups. For the three displacement conditions, there was a strong tendency for the 2.0-mm plate + screw and the grid plate to have higher values.

Evaluación comparativa in vitro de la capacidad de sellado del sistema de conductos radiculares por diferentes técnicas

The aim of this study was to evaluate in vitro the sealing ability of the root canal using different techniques: cold lateral compaction, modified Hybrid Tagger, Thermafil and System B. Material and methods. Were used 116 human single-roots, included in resin blocks, sectioned longitudinally, attached to a device with screws and divided into four groups (n=29). After instrumentation using a bur, depression was made in each third of the root canal in each of the halves. Teeth were filled upon the different techniques. For evaluation of the quality, pictures were taken with a 1.5X magnification, photos and radiographs were utilized. After statistical analysis, were performed both macroscopic and radiographic evaluation, by Chi-square test (χ2). Results. The Thermafil technique showed better results in terms of sealing capacity, failures and homogeneity followed by modified Hybrid Tagger, the System B and cold lateral compaction. Conclusions. Thermafil was the technique that shown to be macroscopic, radiographic and homogeneously best over the other techniques evaluated in regard to the filling of the depressions made (AU)