An in vitro study showing the three-dimensional microenvironment influence over the behavior of head and neck squamous cell carcinoma

Objectives: The Head and Neck Squamous Cell Carcinoma (HNSCC) ranks sixth worldwide. The mechanisms of growth, invasion and metastasis of this pathology are extensively studied and generally related to specific variations in signaling pathways like the PI3K-Akt; however most of these competent studies have been performed bidimensionally, which may hide important questions. This study sought to analyze the influence of the microenvironment upon the behavior of HNSCC. Study Design: The status of pAkt, NF-kappa B and Cyclin D1 proteins was accessed through immunofluorescence and western blot methods in HNSCC cell lines originating from tongue, pharynx and metastatic lymph node when submitted to a three-dimensional culture model utilizing a matrix system. A bidimensional culture model (monolayer) was used as control. Results: The HNSCC cell lines cultured three-dimensionally exhibited a growth pattern characterized by small isolated islands, different from the control group. When the three-dimensional model was applied, two of the studied cell lines showed the same expression pattern as the bidimensional model regarding nuclear or cytoplasmatic localization, as well as reduction of all protein levels; however, the cell line originated from tongue, which specially has the epidermal growth factor receptor constitutively activated, demonstrated nuclear translocation of pAkt and also an increase in the levels of Cyclin D1. Conclusions: The results suggest the influence of the microenvironment upon the behavior of HNSCC cells due to the changed expression of proteins related to tumor growth and cellular invasion. Furthermore, intrinsically genetic conditions also played important roles over the cells, despite the culture model employed.

Cytokine inhibition and time-related influence of inflammatory stimuli on the hyperalgesia induced by the nucleus pulposus

The symptoms of lumbar disc herniation, such as low back pain and sciatica, have been associated with local release of cytokines following the inflammatory process induced by the contact of the nucleus pulposus (NP) with the spinal nerve. Using an animal experimental model of intervertebral disc herniation and behavioral tests to evaluate mechanical (electronic von Frey test) and thermal (Hargreaves Plantar test) hyperalgesia in the hind paw of rats submitted to the surgical model, this study aimed to detect in normal intervertebral disc the cytokines known to be involved in the mechanisms of inflammatory hyperalgesia, to observe if previous exposure of the intervertebral disc tissue to specific antibodies could affect the pain behavior (mechanical and thermal hyperalgesia) induced by the NP, and to observe the influence of the time of contact of the NP with the fifth lumbar dorsal root ganglion (L5-DRG) in the mechanical and thermal hyperalgesia. The cytokines present at highest concentrations in the rat NP were TNF-alpha, IL-1 beta and CINC-1. Rats submitted to the disc herniation experimental model, in which a NP from the sacrococcygeal region is deposited over the right L5-DRG, showed increased mechanical and thermal hyperalgesia that lasted at least 7 weeks. When the autologous NP was treated with antibodies against the three cytokines found at highest concentrations in the NP (TNF-alpha, IL-1 beta and CINC-1), there was decrease in both mechanical and thermal hyperalgesia in different time points, suggesting that each cytokine may be important for the hyperalgesia in different steps of the inflammatory process. The surgical remotion of the NP from herniated rats 1 week after the implantation reduced the hyperalgesia to the level similar to the control group. This reduction in the hyperalgesia was also observed in the group that had the NP removed 3 weeks after the implantation, although the intensity of the hyperalgesia did not decreased totally. The removal of the NP after 5 weeks did not changed the hyperalgesia observed in the hind paw, which suggests that the longer the contact of the NP with the DRG, the greater is the possibility of development of chronic pain. Together our results indicate that specific cytokines released during the inflammatory process induced by the herniated intervertebral disc play fundamental role in the development of the two modalities of hyperalgesia (mechanical and thermal) and that the maintenance of this inflammation may be the most important point for the chronification of the pain.

Influence of obesity and handgrip strength in the static postural balance of active older women

The objective of this study was to investigate the influence of the obesity and handgrip strength on the static balance of active older women in the opened and closed eyes conditions. Thirty one women aged from 65 to 75 years (16 eutrophic and 15 obese) were evaluated. Mean age and BMI of the eutrophic women were, respectively, 68.3 +/- 2.7 years and 23.4 +/- 1.6kg/m(2), and of the obese women were 69.1 +/- 2.7 years and 33.5 +/- 3kg/m(2). Handgrip strength was evaluated using a dynamometer (JAMAR). A tridimensional sensors system was used to evaluate the static postural balance. The tests were performed for 90 seconds, with eyes opened and closed. The mean handgrip strength of the eutrophic women was 25.1 +/- 4.6kgf and of the obese women was 24.8 +/- 5.2kgf, (p>0,05). Significant differences between groups were only observed in the maximum displacement with opened eyes (p=0,04) and closed eyes(p<0,01). There was no correlation between the maximum displacement neither with the BMI or the handgrip strength. The present study showed smaller a-p displacement in obese than in eutrophic women, with major statistic difference in the eyes closed condition. In the present study, the handgrip strength did not influence the static balance, however the obesity was a determinant factor for the smaller a-p displacement of the active older women.

Effect of the thermostat in the molecular dynamics simulation on the folding of the model protein chignolin

Molecular dynamics simulations of the model protein chignolin with explicit solvent were carried out, in order to analyze the influence of the Berendsen thermostat on the evolution and folding of the peptide. The dependence of the peptide behavior on temperature was tested with the commonly employed thermostat scheme consisting of one thermostat for the protein and another for the solvent. The thermostat coupling time of the protein was increased to infinity, when the protein is not in direct contact with the thermal bath, a situation known as minimally invasive thermostat. In agreement with other works, it was observed that only in the last situation the instantaneous temperature of the model protein obeys a canonical distribution. As for the folding studies, it was shown that, in the applications of the commonly utilized thermostat schemes, the systems are trapped in local minima regions from which it has difficulty escaping. With the minimally invasive thermostat the time that the protein needs to fold was reduced by two to three times. These results show that the obstacles to the evolution of the extended peptide to the folded structure can be overcome when the temperature of the peptide is not directly controlled.

The influence of captive breeding on the fatty acid profiles of Salminus hilarii (Characiformes: Characidae) eggs and larvae

In addition to the strong influence of the broodstock diet on the development and survival of offspring, domestication may also interfere with the larval life success. We obtained eggs from wild and domesticated Salminus hilarii females and domesticated males. Wild females were caught in the Tiete River and tributaries, and the domesticated females were born three years before the beginning of the experiment in the Ponte Nova Fish Farm. Animals from both groups were fed with the same feed to exclude feed variables. The eggs and larvae were sampled at 0, 8, 16, and 28 h after spawning (HAS), with the last sampling (28 HAS) coinciding with hatching time. After hatching, samplings proceeded at 32, 48, 66, and 96 HAS, with the last sampling (96 HAS) corresponding to the end of yolk sac consumption. Finally, the last experimental period was during the larvae exogenous feeding phase, at 102, 118, 166, and 214 HAS. Our data revealed that domestication of S. hilarii females influenced fatty acid (FA) metabolism during embryo and larva development. However, the structure of membrane phospholipid FA remained mostly stable, with changes principally in the neutral fraction. When the external conditions, mainly water and feed quality, remained constant, domestication of S. hilarii females did not significantly affect the structural FA composition but influenced the selectivity of consumption and/or storage of specific FA.

Influence of HSM cutting parameters on the surface integrity characteristics of hardened AISI H13 steel

The purpose of this study is to evaluate the influence of the cutting parameters of high-speed machining milling on the characteristics of the surface integrity of hardened AISI H13 steel. High-speed machining has been used intensively in the mold and dies industry. The cutting parameters used as input variables were cutting speed (v c), depth of cut (a p), working engagement (a e) and feed per tooth (f z ), while the output variables were three-dimensional (3D) workpiece roughness parameters, surface and cross section microhardness, residual stress and white layer thickness. The subsurface layers were examined by scanning electron and optical microscopy. Cross section hardness was measured with an instrumented microhardness tester. Residual stress was measured by the X-ray diffraction method. From a statistical standpoint (the main effects of the input parameters were evaluated by analysis of variance), working engagement (a e) was the cutting parameter that exerted the strongest effect on most of the 3D roughness parameters. Feed per tooth (f z ) was the most important cutting parameter in cavity formation. Cutting speed (v c) and depth of cut (a p) did not significantly affect the 3D roughness parameters. Cutting speed showed the strongest influence on residual stress, while depth of cut exerted the strongest effect on the formation of white layer and on the increase in surface hardness.

Testing the influence of far-field topographic forcing on subduction initiation at a passive margin

Despite favourable gravitational instability and ridge-push, elastic and frictional forces prevent subduction initiation fromarising spontaneously at passive margins. Here,we argue that forces arising fromlarge continental topographic gradients are required to initiate subduction at passivemargins. In order to test this hypothesis,we use 2Dnumerical models to assess the influence of the Andean Plateau on stressmagnitudes and deformation patterns at the Brazilian passive margin. The numerical results indicate that “plateau-push” in this region is a necessary additional force to initiate subduction. As the SE Brazilianmargin currently shows no signs of self-sustained subduction, we examined geological and geophysical data to determine if themargin is in the preliminary stages of subduction initiation. The compiled data indicate that the margin is presently undergoing tectonic inversion, which we infer as part of the continental–oceanic overthrusting stage of subduction initiation. We refer to this early subduction stage as the “Brazilian Stage”, which is characterized by N10 kmdeep reverse fault seismicity at themargin, recent topographic uplift on the continental side, thick continental crust at themargin, and bulging on the oceanic side due to loading by the overthrusting continent. The combined results of the numerical simulations and passivemargin analysis indicate that the SE Brazilian margin is a prototype candidate for subduction initiation.