Histological and three-dimensional evaluation of osseointegration to nanostructured calcium phosphate-coated implants

Nanostructures on implant surfaces have been shown to enhance osseointegration; however, commonly used evaluation techniques are probably not sufficiently sensitive to fully determine the effects of this process. This study aimed to observe the osseointegration properties of nanostructured calcium phosphate (CaP)-coated implants, by using a combination of three-dimensional imaging and conventional histology. Titanium implants were coated with stable CaP nanoparticles using an immersion technique followed by heat treatment. Uncoated implants were used as the control. After topographical and chemical characterizations, implants were inserted into the rabbit femur. After 2 and 4 weeks, the samples were retrieved for micro-computed tomography and histomorphometric evaluation. Scanning electron microscopy evaluation indicated that the implant surface was modified at the nanoscale by CaP to obtain surface textured with rod-shaped structures. Relative to the control, the bone-to-implant contact for the CaP-coated implant was significantly higher at 4 weeks after the implant surgery. Further, corresponding 3-D images showed active bone formation surrounding the implant. 3-D quantification and 2-D histology demonstrated statistical correlation; moreover, 3-D quantification indicated a statistical decrease in bone density in the non-coated control implant group between 2 and 4 weeks after the surgery. The application of 3-D evaluation further clarified the temporal characteristics and biological reaction of implants in bone. (C) 2011 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Desenvolvimento da aveia branca e disponibilidade de fósforo em razão da aplicação de silicato de potássio

A aplicação de silicato de potássio pode aumentar a disponibilidade de fósforo no solo, além de influenciar no crescimento da aveia branca. Objetivou-se neste trabalho avaliar o efeito da aplicação de doses de silicato de potássio no solo no desenvolvimento da aveia branca e na disponibilidade de fósforo. O delineamento experimental foi inteiramente ao acaso, em esquema fatorial 2 x 4, com seis repetições. Os tratamentos foram 20 e 200 mg dm-3 de P2O5 combinados com 0, 150, 300 e 450 mg dm-3 de Si, aplicados por ocasião da semeadura. Após a colheita, determinaram-se a matéria seca de folhas + colmos, das panículas e o total da parte aérea. A palha das panículas foi separada para avaliação dos teores de P e Si nas folhas + colmos, grãos e palha das panículas. As concentrações de Si e P no solo foram avaliadas após colheita das plantas. A aplicação de silicato aumenta os teores de Si no solo e influencia na maior disponibilidade de fósforo. A matéria seca de folhas + colmos é incrementada pelo silicato quando os teores iniciais de P são baixos. Mesmo com a dose elevada de P, a aplicação de silicato aumenta a disponibilidade de P no solo e a absorção pelas plantas. Os teores de Si na palha das panículas, folhas + colmos e grãos são influenciados pela maior disponibilidade do elemento no solo.

IMPROVEMENT TO THE MARGINAL COPING FIT of COMMERCIALLY PURE TITANIUM CAST IN PHOSPHATE-BONDED INVESTMENT BY USING A SIMPLE PATTERN COATING TECHNIQUE

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

Biological Response to Wollastonite Doped alpha-Tricalcium Phosphate Implants in Hard and Soft Tissues in Rats

The biological response following subcutaneous and bone implantation of beta-wollastonite(beta-W)-doped alpha-tricalcium phosphate bioceramics in rats was evaluated. Tested materials were: tricalciurn phosphate (TCP), consisting of a mixture of alpha- and beta-polymorphs; TCP doped with 5 wt. % of beta-W (TCP5W), composed of alpha-TCP as only crystalline phase; and TCP doped with 15 wt. % of beta-W (TCP15), containing crystalline alpha-TCP and beta-W. Cylinders of 2x1 mm were implanted in tibiae and backs of adult male Rattus norvegicus, Holtzman rats. After 7, 30 and 120 days, animals were sacrificed and the tissue blocks containing the implants were excised, fixed and processed for histological examination. TCP, TCP5W and TCP15W implants were biocompatible but neither bioactive nor biodegradable in rat subcutaneous tissue. They were not osteoinductive in connective tissue either. However, in rat bone tissue beta-W-doped alpha-TCP implants (TCP5W and TCP15W) were bioactive, biodegradable and osteoconductive. The rates of biodegradation and new bone formation observed for TCP5W and TCP15W implants in rat bone tissue were greater than for non-doped TCP.

Effects of indoramin in rat vas deferens and aorta: concomitant alpha(1)-adrenoceptor and neuronal uptake blockade

1 the actions of the alpha(1)-adrenoceptor antagonist indoramin have been examined against the contractions induced by noradrenaline in the rat vas deferens and aorta taking into account a putative neuronal uptake blocking activity of this antagonist which could. result in self-cancelling actions.2 Indoramin behaved as a simple competitive antagonist of the contractions induced by noradrenaline in the vas deferens and aorta yielding pA(2) values of 7.38 +/- 0.05 (slope = 0.98 +/- 0.03) and 6.78 +/- 0.14 (slope = 1.08 +/- 0.06), respectively.3 When the experiments were repeated in the presence of cocaine (6 mu M) the potency (pA(2)) of indoramin in antagonizing the contractions of the vas deferens to noradrenaline was increased to 8.72 +/- 0.07 (slope = 1.10 +/- 0.05) while its potency remained unchanged in the aorta (pA(2) = 6.69 +/- 0.12; slope = 1.04 +/- 0.05).4 In denervated vas deferens, indoramin antagonized the contractions to noradrenaline with a potency similar to that found in the presence of cocaine (8.79 +/- 0.07; slope = 1.09 +/- 0.06).5 It is suggested that indoramin blocks alpha(1)-adrenoceptors and neuronal uptake in rat vas deferens resulting in Schild plots with slopes not different from unity even in the absence of selective inhibition of neuronal uptake. As a major consequence of this double mechanism of action, the pA(2) values for this antagonist are underestimated when calculated in situations where the neuronal uptake is active, yielding spurious pK(B) values.

In vivo uptake of a haem analogue Zn protoporphyrin IX by the human malaria parasite P. falciparum-infected red blood cells

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

Phosphate distribution in the sediments along a cascade of reservoirs - (Paranapanema River, SE, Brazil)

Total phosphorus concentrations in the sediment of a cascade of 8 reservoirs located in the Paranapanema River (SE, Brazil) were analysed during two consecutive seasonal periods (2000 and 200 1). The reservoirs of Jurumirim, Chavantes, Salto Grande, Canoas 11, Canoas 1, Capivara, Taquarucu and Rosana were built during the last five decades with the purpose of hydroelectric generation. The sampling points were distributed along a 700 km of river stretch in 19 stations distributed in the river-reservoirs zones and main tributaries as well as in 5 stations representing the adjacent wetlands. The nutrient concentration was determined in samples from surface layers of sediment. Temporal and spatial changes were observed for both years. Three different areas could be identified along the river, considering not only Tot-P variations but also others physical and chemical characteristics such as organic matter concentration, nitrogen concentration of sediment and 02, pH, conductivity and Eh measured in the bottom layer of the water column. Higher values of Tot-P were found in the middle and lower region of the Paranapanema basin (maximum value of 1.96 mg g(-1) in Tibagi River in October/2001). Agricultural practices and cattle raising are the main external sources of nutrient in these regions. Superficial or sub-superficial watershed drainage and tributaries entrances are important nutrient contributors, leading to an increasing accumulation of nutrient in the sediment. This process seems to be especially important in the middle region of the Paranapanema Valley, were the values of Tot-P were higher comparing to the other areas.

Dynamics of glyphosate-induced conformational changes of Mycobacterium tuberculosis 5-enolpyruvylshikimate-3-phosphate synthase (EC 2.5.1.19) determined by hydrogen-deuterium exchange and electrospray mass spectrometry

The enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) catalyzes the reaction between shikimate 3-phosphate and phosphoenolpyruvate to form 5-enolpyruvylshikimate 3-phosphate, an intermediate in the shikimate pathway, which leads to the biosynthesis of aromatic amino acids. EPSPS exists in an open conformation in the absence of substrates and/or inhibitors and in a closed conformation when bound to the substrate and/or inhibitor. In the present report, the H/D exchange properties of EPSPS from Mycobacterium tuberculosis (Mt) were investigated for both enzyme conformations using ESI mass spectrometry and circular dichroism (CD). When the conformational changes identified by H/D exchanges were mapped on the 3-D structure, it was observed that the apoenzyme underwent extensive conformational changes due to glyphosate complexation, characterized by an increase in the content of alpha-helices from 40% to 57%, while the beta-sheet content decreased from 30% to 23%. These results indicate that the enzyme underwent a series of rearrangements of its secondary structure that were accompanied by a large decrease in solvent access to many different regions of the protein. This was attributed to the compaction of 71% of alpha-helices and 57% of beta-sheets as a consequence of glyphosate binding to the enzyme. Apparently, MtEPSPS undergoes a series of inhibitor-induced conformational changes, which seem to have caused synergistic effects in preventing solvent access to the core of molecule, especially in the cleft region. This may be part of the mechanism of inhibition of the enzyme, which is required to prevent the hydration of the substrate binding site and also to induce the cleft closure to avoid entrance of the substrates.

Hydrogen/deuterium exchange mass spectrometry for characterizing phosphoenolpyruvate-induced structural transitions in Mycobacterium tuberculosis 5-enolpyruvylshikimate-3-phosphate synthase (EC 2.5.1.1)

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

Phosphate closes the solution structure of the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) from Mycobacterium tuberculosis

The 5-enolpyruvylshikimate-3-phosphate synthase catalyses the sixth step of the shikimate pathway that is responsible for synthesizing aromatic compounds and is absent in mammals, which makes it a potential target for drugs development against microbial diseases. Here, we report the phosphate binding effects at the structure of the 5-enolpyruvyl shikimate-3-phosphate synthase from Mycobacterium tuberculosis. This enzyme is formed by two similar domains that close on each other induced by ligand binding, showing the occurrence of a large conformation change. We have monitored the phosphate binding effects using analytical ultracentrifugation, small angle X-ray scattering and, circular dichroism techniques. The low resolution results showed that the enzyme in the presence of phosphate clearly presented a more compact structure. Thermal-induced unfolding experiments followed by circular dichroism suggested that phosphate rigidified the enzyme. Summarizing, these data suggested that the phosphate itself is able to induce conformational change resulting in the closure movement in the M. tuberculosis 5-enolpyruvylshikimate-3-phosphate synthase. (c) 2006 Elsevier B.V. All rights reserved.

Muscle damage slows oxygen uptake kinetics during moderate-intensity exercise performed at high pedal rate

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

The highest intensity and the shortest duration permitting attainment of maximal oxygen uptake during cycling: effects of different methods and aerobic fitness level

The aims of this study were: (1) to verify the validity of previous proposed models to estimate the lowest exercise duration (T (LOW)) and the highest intensity (I (HIGH)) at which VO(2)max is reached (2) to test the hypothesis that parameters involved in these models, and hence the validity of these models are affected by aerobic training status. Thirteen cyclists (EC), eleven runners (ER) and ten untrained (U) subjects performed several cycle-ergometer exercise tests to fatigue in order to determine and estimate T (LOW) (ET (LOW)) and I (HIGH) (EI (HIGH)). The relationship between the time to achieved VO(2)max and time to exhaustion (T (lim)) was used to estimate ET (LOW). EI (HIGH) was estimated using the critical power model. I (HIGH) was assumed as the highest intensity at which VO2 was equal or higher than the average of VO(2)max values minus one typical error. T (LOW) was considered T (lim) associated with I (HIGH). No differences were found in T (LOW) between ER (170 +/- 31 s) and U (209 +/- 29 s), however, both showed higher values than EC (117 +/- 29 s). I (HIGH) was similar between U (269 +/- 73 W) and ER (319 +/- 50 W), and both were lower than EC (451 +/- 33 W). EI (HIGH) was similar and significantly correlated with I-HIGH only in U (r = 0.87) and ER (r = 0.62). ET (LOW) and T (LOW) were different only for U and not significantly correlated in all groups. These data suggest that the aerobic training status affects the validity of the proposed models for estimating I (HIGH).

Allosteric water and phosphate effects in Hoplosternum littorale hemoglobins

This paper reports the results obtained using the osmotic stress method applied to the purified cathodic and anodic hemoglobins (Hbs) from the catfish Hoplosternum littorale, a species that displays facultative accessorial air oxygenation. We demonstrate that water potential affects the oxygen affinity of H. littorale Hbs in the presence of an inert solute (sucrose). Oxygen affinity increases when water activity increases, indicating that water molecules stabilize the high-affinity state of the Hb. This effect is the same as that observed in tetrameric vertebrate Hbs. We show that both anodic and cathodic Hbs show conformational substrates similar to other vertebrate Hbs. For both Hbs, addition of anionic effectors, especially chloride, strongly increases the number of water molecules bound, although anodic Hb did not exhibit sensitivity to saturating levels of ATP. Accordingly, for both Hbs, we propose that the deoxy conformations coexist in at least two anion-dependent allosteric states, T-o and T-x, as occurs for human Hb. We found a single phosphate binding site for the cathodic Hb.