Model peptides mimic the structure and function of the N-terminus of the pore-forming toxin sticholysin II

To investigate the role of the N-terminal region in the lytic mechanism of the pore-forming toxin sticholysin II (St II), we studied the conformational and functional properties of peptides encompassing the first 30 residues of the protein. Peptides containing residues 1-30 (P1-30) and 11-30 (P11-30) were synthesized and their conformational properties were examined in aqueous solution as a function of peptide concentration, pH, ionic strength, and addition of the secondary structure-inducing solvent trifluoroethanol (TFE). CD spectra showed that increasing concentration, pH, and ionic strength led to aggregation of P1-30; as a consequence, the peptide acquired beta-sheet conformation. In contrast, P11-30 exhibited practically no conformational changes under the same conditions, remaining essentially structureless. Moreover, this peptide did not undergo aggregation. These differences clearly point to the modulating effect of the first 10 hydrophobic residues on the peptides aggregation and conformational properties. In TFE both the first ten hydrophobic peptides acquired alpha-helical conformation, albeit to a different extent, P11-30 displayed lower alpha-helical content. P1-30 presented a larger-fraction of residues in alpha-helical conformation in TFE than that found in St II's crystal structure for that portion of the protein. Since TFE mimics the membrane em,, such increase in helical content could also occur upon toxin binding to membranes and represent a step in the mechanism of pore formation. The peptides conformational properties correlated well with their functional behaviour. Thus, P1-30 exhibited much higher hemolytic activity than P11-30. In addition, P11-30 was able to block the toxin's hemolytic activity. The size of pores formed in red blood cells by P 1-30 was estimated by measuring the permeability PEGs of different molecular mass. The pore radius (0.95 +/- 0.01 nm) was very similar to that of the PEGs of different pore formed by the toxin. The results demonstrate that the synthetic peptide P1-30 is a good model of St 11 conformation and function and emphasize the contribution of the toxin's N-terminal region, and, in particular, the hydrophobic residues 1-10 to pore formation. (c) 2005 Wiley Periodicals, Inc.

Mechanical strength and subcritical crack growth under wet cyclic loading of glass-infiltrated dental ceramics

Objectives. Evaluate the flexural strength (sigma) and subcritical crack growth (SCG) under cyclic loading of glass-infiltrated alumina-based (IA, In-Ceram Alumina) and zirconia-reinforced (IZ, In-Ceram Zirconia) ceramics, testing the hypothesis that wet environment influences the SCG of both ceramics when submitted to cyclic loading.Methods. Bar-shaped specimens of IA (n = 45) and IZ ( n = 45) were fabricated and loaded in three-point bending (3P) in 37 degrees C artificial saliva (IA(3P) and IZ(3P)) and cyclic fatigued (F) in dry (D) and wet (W) conditions (IA(FD), IA(FW), IZ(FD), IZ(FW)). The initial sigma and the number of cycles to fracture were obtained from 3P and F tests, respectively. Data was examined using Weibull statistics. The SCG behavior was described in terms of crack velocity as a function of maximum stress intensity factor (K(Imax)).Results. The Weibull moduli (m = 8) were similar for both ceramics. The characteristic strength (sigma(0)) of IA and IZ was and 466 MPa 550 MPa, respectively. The wet environment significantly increased the SCG of IZ, whereas a less evident effect was observed for IA. In general, both ceramics were prone to SCG, with crack propagation occurring at K(I) as low as 43-48% of their critical K(I). The highest sigma of IZ should lead to longer lifetimes for similar loading conditions.Significance. Water combined with cyclic loading causes pronounced SCG in IZ and IA materials. The lifetime of dental restorations based on these ceramics is expected to increase by reducing their direct exposure to wet conditions and/or by using high content zirconia ceramics with higher strength. (C) 2010 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Burrows with Chimneys of the Fiddler Crab Uca thayeri: Construction, Occurrence, and Function

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

Enhancement of thermal stability, strength and extensibility of lipid-based polyurethanes with cellulose-based nanofibers

Cellulose was extracted from lignocellulosic fibers and nanocrystalline cellulose (NC) prepared by alkali treatment of the fiber, steam explosion of the mercerized fiber, bleaching of the steam exploded fiber and finally acid treatment by 5% oxalic acid followed again by steam explosion. The average length and diameter of the NC were between 200-250 nm and 4-5 nm, respectively, in a monodisperse distribution. Different concentrations of the NC (0.1, 0.5, 1.0, 1.5, 2.0 and 2.5% by weight) were dispersed non-covalently into a completely bio-based thermoplastic polyurethane (TPU) derived entirely from oleic acid. The physical properties of the TPU nanocomposites were assessed by Fourier Transform Infra-Red spectroscopy (FTIR), Thermo-Gravimetric Analysis (TGA), Differential Scanning Calorimetry (DSC), X-Ray Diffraction (XRD), Dynamic Mechanical Analysis (DMA) and Mechanical Properties Analysis. The nanocomposites demonstrated enhanced stress and elongation at break and improved thermal stability compared to the neat TPU. The best results were obtained with 0.5% of NC in the TPU. The elongation at break of this sample was improved from 178% to 269% and its stress at break from 29.3 to 40.5 MPa. In this and all other samples the glass transition temperature, melting temperature and crystallization behavior were essentially unaffected. This finding suggests a potential method of increasing the strength and the elongation at break of typically brittle and weak lipid-based TPUs without alteration of the other physico-chemical properties of the polymer. (C) 2012 Elsevier Ltd. All rights reserved.

Aggregates in the strength and gravity regime: Particles sizes in Saturn's rings

Particles in Saturn's main rings range in size from dust to kilometer-sized objects. Their size distribution is thought to be a result of competing accretion and fragmentation processes. While growth is naturally limited in tidal environments, frequent collisions among these objects may contribute to both accretion and fragmentation. As ring particles are primarily made of water ice attractive surface forces like adhesion could significantly influence these processes, finally determining the resulting size distribution. Here, we derive analytic expressions for the specific self-energy Q and related specific break-up energy Q(star) of aggregates. These expressions can be used for any aggregate type composed of monomeric constituents. We compare these expressions to numerical experiments where we create aggregates of various types including: regular packings like the face-centered cubic (fcc), Ballistic Particle Cluster Aggregates (BPCA), and modified BPCAs including e.g. different constituent size distributions. We show that accounting for attractive surface forces such as adhesion a simple approach is able to: (a) generally account for the size dependence of the specific break-up energy for fragmentation to occur reported in the literature, namely the division into "strength" and "gravity" regimes and (b) estimate the maximum aggregate size in a collisional ensemble to be on the order of a few tens of meters, consistent with the maximum particle size observed in Saturn's rings of about 10 m. (c) 2012 Elsevier B.V. All rights reserved.

Alterations in the intestinal wall due to protein malnutrition in rats. Evaluation of the rupture strength and the tissue's collagen

OBJETIVO: Avaliar o efeito da desnutrição protéica na parede intestinal do rato através da medida de força de ruptura e dosagem do colágeno tecidual no íleo e cólon distal. MÉTODOS: Foram utilizados 120 ratos, pesando em média 100g, que receberam durante 07 dias uma dieta padrão, contendo 20% de caseína para adaptação dos animais as condições do biotério. Após esse período os animais foram divididos em dois grupos de 60, o controle denominado grupo um que recebeu a dieta padrão, e o grupo teste denominado grupo dois, que recebeu dieta hipoprotéica contendo 2% de caseína. Os dois grupos receberam suas respectivas dietas por um período de 21 dias. Após esse período iniciou-se o sacrifício seqüencial dos animais em ambos os grupos, em número de 12 animais em cada momento, correspondendo ao dia Zero (MO), 4º dia (M1), 7º dia (M2), 14º dia (M3), e 21º dia (M4) sendo mantida a mesma dieta até o final do sacrifício. em cada momento foram avaliados o peso corpóreo, albumina sanguínea, hidroxiprolina tecidual, relação hidroxiprolina/proteína tecidual e a força de ruptura no segmento ileal e cólico dos animais. RESULTADOS: Observou-se que a força de ruptura do segmento ileal e do cólon distal foi menor nos animais desnutridos (Grupo 2). A perda da resistência mecânica foi maior no segmento do cólon distal do que no segmento ileal, provavelmente pela menor concentração do colágeno tecidual no cólon distal. CONCLUSÃO: A desnutrição protéica induz a diminuição da resistência mecânica no íleo e no cólon distal associado a diminuição do colágeno tecidual na parede intestinal.

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.

Loss of Expression and Function of SOCS3 Is an Early Event in HNSCC: Altered Subcellular Localization as a Possible Mechanism Involved in Proliferation, Migration and Invasion

Background: Suppressor of cytokine signaling 3 (SOCS3) is an inducible endogenous negative regulator of signal transduction and activator of transcription 3 (STAT3). Epigenetic silencing of SOCS3 has been shown in head and neck squamous cell carcinoma (HNSCC), which is associated with increased activation of STAT3. There is scarce information on the functional role of the reduction of SOCS3 expression and no information on altered subcellular localization of SOCS3 in HNSCC.Methodology/Principal Findings: We assessed endogenous SOCS3 expression in different HNSCC cell lines by RT-qPCR and western blot. Immunofluorescence and western blot were used to study the subcellular localization of endogenous SOCS3 induced by IL-6. Overexpression of SOCS3 by CMV-driven plasmids and siRNA-mediated inhibition of endogenous SOCS3 were used to verify the role of SOCS3 on tumor cell proliferation, viability, invasion and migration in vitro. In vivo relevance of SOCS3 expression in HNSCC was studied by quantitative immunohistochemistry of commercially-available tissue microarrays. Endogenous expression of SOCS3 was heterogeneous in four HNSCC cell lines and surprisingly preserved in most of these cell lines. Subcellular localization of endogenous SOCS3 in the HNSCC cell lines was predominantly nuclear as opposed to cytoplasmic in non-neoplasic epithelial cells. Overexpression of SOCS3 produced a relative increase of the protein in the cytoplasmic compartment and significantly inhibited proliferation, migration and invasion, whereas inhibition of endogenous nuclear SOCS3 did not affect these events. Analysis of tissue microarrays indicated that loss of SOCS3 is an early event in HNSCC and was correlated with tumor size and histological grade of dysplasia, but a considerable proportion of cases presented detectable expression of SOCS3.Conclusion: Our data support a role for SOCS3 as a tumor suppressor gene in HNSCC with relevance on proliferation and invasion processes and suggests that abnormal subcellular localization impairs SOCS3 function in HNSCC cells.

Diametral tensile strength and film thickness of an experimental dental luting agent derived from castor oil

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

Flexural strength and hardness of reline and denture base acrylic resins after different exposure times of microwave disinfection

Objectives: To evaluate the effect of microwave disinfection on the flexural strength and Vickers hardness of 4 autopolymerized resins (Kooliner [K], Tokuso Rebase Fast [T], Ufi Gel Hard [U], and New Truliner [N]) and 1 denture base resin (Lucitone 550 [L]). Method and Materials: For each material, 48 specimens (64 x 10 x 3.3 mm) were made and divided into 6 equal groups (n = 8). In the control group, specimens were untreated. Before testing, specimens were immersed in 200 mL of distilled water and submitted to disinfection for 1 of the following irradiation times: 1, 2, 3, 4, or 5 minutes. The irradiation procedure was performed twice. The flexural strength was determined using a testing machine MTS-810 and measurements of Vickers hardness were made on Micromet 2100. The values were submitted to ANOVA and Tukey's test (P = .05). Results: The K material showed a significant increase (P = .0010) in flexural strength following 5 minutes of disinfection compared to control specimens. The flexural strength mean values of materials T, U, and N were not significantly affected (P > .05) by disinfection. Compared to the control group, the K material showed a significant increase in hardness (P < .001) following disinfection for 3, 4, and 5 minutes. For material U, disinfection for 4 and 5 minutes produced specimens with significantly increased hardness values (P < .001) compared to the control group. For material N, disinfection for 5 minutes resulted in significantly higher hardness values (P < .001) than the control group. Conclusion: Regardless of the irradiation time, the flexural strength and hardness of the materials evaluated were not detrimentally affected by microwave disinfection. (Quintessence Int 2008;39:833-840)

Effect of water storage on the shear strength and fatigue limit of the reline resin bond to denture base resins

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

Mechanical Strength and Analysis of Fracture of Titanium Joining Submitted to Laser and Tig Welding

This study compared the tensile strength and fracture mechanism of tungsten inert gas (TIG) welds in cylindrical rods of commercially pure titanium (cp Ti) with those of laser welds and intact samples. Thirty dumbbell-shaped samples were developed by using brass rods as patterns. The samples were invested in casings, subjected to thermal cycles, and positioned in a plasma arc welding machine under argon atmosphere and vacuum, and titanium was injected under vacuum/pressure. The samples were X-rayed to detect possible welding flaws and randomly assigned to three groups to test the tensile strength and the fracture mechanism: intact, laser welding, and TIG welding. The tensile test results were investigated using ANOVA, which indicated that the samples were statistically similar. The fracture analysis showed that the cpTi samples subjected to laser welding exhibited brittle fracture and those subjected to TIG welding exhibited mixed brittle/ductile fracture with a predominance of ductile fracture with the presence of microcavities and cleavage areas. Intact samples presented the characteristic straightening in the fracture areas, indicating the ductility of the material.

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.

Reproductive development and function of female rats exposed to di-eta-butyl-phthalate (DBP) in utero and during lactation

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