Morphological characterization by SEM, TEM and AFM of nanoparticles and functional nanocomposites based on natural rubber filled with oxide nanopowders

Nanocomposites were prepared from mixture of different concentrations of ferroelectric nanoparticles in an elastomeric matrix based on the vulcanized natural rubber. The morphological characterization of nanocomposites was carried out using Scanning electron microscopy (SEM), Transmission electron microscopy (TEM) and Atomic force microscopy (AFM). The nanocrystalline ferroelectric oxide is potassium strontium niobate (KSN) with stoichiometry KSr2Nb5O15, and was synthesized by the chemical route using a modified polyol method, obtaining particle size and microstrain equal to 20 nm and 0.32, respectively. These ferroelectric nanoparticles were added into the natural rubber in concentrations equal to 1, 3, 5, 10, 20 and 50 phr (parts per hundred of rubber) forming ferroelectric nanocomposites (NR/KSN). Using morphological characterization, we identified the maximum value of surface roughness at low concentrations, in particular, sample with 3 phr of nanoparticles and factors such as encapsulation and uniformity in the distribution of nanoparticles into the natural rubber matrix are investigated and discussed.

Myoglobin from the burrowing reptile Amphisbaena alba: concentrations and functional characteristics

1. 1. Myoglobin from the subterranean reptile Amphisbaena alba was isolated for measurement of concentrations and physico-chemical properties. 2. 2. The concentrations (averaging 12.1 mg.g-1 wet weight in the temporal muscles and 5.8-6.0 in the muscles that motivate the wedge-shaped head which forms the burrowing tool) far exceed those earlier reported for reptiles and other terrestrial vertebrates. 3. 3. The myoglobin has a low O2 affinity compared to mammals (P50 = 2mmHg at 25°C). In the presence of the same myoglobin O2 tension as in mammals this appears to favour similar in vivo O2 saturations at the lower reptilian body temperature. 4. 4. The temperature sensitivity of P50 reflect a heat of oxygenation, ΔH near -13 kcal· mol-1. The myoglobin is monomeric and thus lacks cooperativity in O2 binding and there is no Bohr effect. 5. 5. The pattern of microheterogeneity is similar to that of myoglobin of terrestrial vertebrates but different to aquatic mammals and reptiles. The major and two minor components exhibit very similar O2 affinities. 6. 6. The concentrations and oxygen-binding characteristics of Amphisbaena myoglobin are discussed with regard to the flow of O2 to the mitochondria during digging activity in hypoxic burrow environments. © 1981.

Apoptosis and reduced microvascular density of the lamina propria during tooth eruption in rats

During tooth eruption, structural and functional changes must occur in the lamina propria to establish the eruptive pathway. In this study, we evaluate the structural changes that occur during lamina propria degradation and focus these efforts on apoptosis and microvascular density. Fragments of maxilla containing the first molars from 9-, 11-, 13- and 16-day-old rats were fixed, decalcified and embedded in paraffin. The immunohistochemical detection of vascular endothelial growth factor (VEGF), caspase-3 and MAC387 (macrophage marker), and the TUNEL method were applied to the histological molar sections. The numerical density of TUNEL-positive cells and VEGF-positive blood vessel profiles were also obtained. Data were statistically evaluated using a one-way anova with the post-hoc Kruskal-Wallis or Tukey test and a significance level of P ≤ 0.05. Fragments of maxilla were embedded in Araldite for analysis under transmission electron microscopy (TEM). TUNEL-positive structures, fibroblasts with strongly basophilic nuclei and macrophages were observed in the lamina propria at all ages. Using TEM, we identified processes of fibroblasts or macrophages surrounding partially apoptotic cells. We found a high number of apoptotic cells in 11-, 13- and 16-day-old rats. We observed VEGF-positive blood vessel profiles at all ages, but a significant decrease in the numerical density was found in 13- and 16-day-old rats compared with 9-day-old rats. Therefore, the establishment of the eruptive pathway during the mucosal penetration stage depends on cell death by apoptosis, the phagocytic activity of fibroblasts and macrophages, and a decrease in the microvasculature due to vascular cell death. These data point to the importance of vascular rearrangement and vascular neoformation during tooth eruption and the development of oral mucosa.

Influence of Mg on the structural and optical properties of LiNbO3 thin films grown by polymeric precursor method

A polymeric precursor solution was used to deposit pure and Mg doped LiNbO3 thin films on sapphire substrates by spin-coating. The effects of magnesium addition on crystallinity, morphology and optical properties of the annealed films were investigated. X-ray diffraction patterns indicate the oriented growth of the films. Phi-scan diffraction evidenced the epitaxial growth with two in-plane variants. AFM studies show that the films are very homogeneous, dense and present smooth surfaces. The refractive index and optical losses obtained by the prism coupling method were influenced by the magnesium addition.

Oral focal mucinosis: clinical case presentation with a focus on aesthetic and functional surgical resolution

Introduction: When planning excisional surgery, alterations in the oral mucosa, particularly in esthetically important areas, represent a challenge. For cases involving an exophytic lesion with a vegetative tumoral aspect, a clinician may prioritize obtaining a definitive diagnosis by histopathologic and histochemical examinations over the preservation of esthetics or maintenance of the involved tooth. Case Presentation: A painless, hard lesion with normal coloration was present in the maxillary left central incisor of a patient for at least 6 years. A biopsy was performed to preserve oral mucosa and a treatment plan was established that included endodontic retreatment and prosthetic restoration. The histopathologic diagnosis was oral focal mucinosis, an uncommon and poorly characterized type of lesion. Conclusion: A conservative biopsy of the lesion facilitated an accurate diagnosis, promoted the patient’s comfort, and achieved satisfactory esthetic results.

Structural and Magnetic Properties of Neodymium - Iron - Boron Clusters

Using inert gas condensation techniques the properties of sputtered neodymium-iron-born clusters were investigated. A D.C. magnetron sputtering source created vaporous Nd-Fe-B which was then condensed into clusters and deposited onto silicon substrates. A composite target of Nd-Fe-B discs on an iron plate and a composite target of Nd-(Fe-Co)-B were utilized to create clusters. The clusters were coated with a carbon layer through R.F. sputtering to prevent oxidation. Samples were investigated in the TEM and showed a size distribution with an average particle diameter of 8.11 nm. The clusters, upon deposition, were amorphous as indicated by diffuse diffraction patterns obtained through SAD. The EDS showed compositionally a direct correlation in the ratio of rare-earth to transition metals between the target and deposited samples. The magnetic properties of the as-deposited clusters showed superparamagnetic properties at high temperatures and ferromagnetic properties at low temperatures; these properties are indicative of rare-earth transition metal amorphous clusters. Annealing of samples showed an initial increase in the coercivity. Samples were annealed in an inert gas atmosphere at 600o C for increasing amounts of time. The samples showed an initial increase in coercivity, but showed no additional increases with additional annealing time. SAD of annealed cluster samples showed the presence of Nd2Fe17 and a bcc-Nd phase. The bcc-Nd is the result of oxidation at high temperatures created during annealing and surface interface energy. The magnetic properties of the annealed samples showed weak coercivity and a saturation magnetization equivalent to that of Nd2Fe17. The annealed clusters showed a slight increase in coercivity at low temperatures. These results indicate a loss of boron during the sputtering process.

Acid-base and biochemical stabilization and quality of recovery in male cats with urethral obstruction and anesthetized with propofol or a combination of ketamine and diazepam

This study compared acid-base and biochemical changes and quality of recovery in male cats with experimentally induced urethral obstruction and anesthetized with either propofol or a combination of ketamine and diazepam for urethral catheterization. Ten male cats with urethral obstruction were enrolled for urethral catheterization and anesthetized with either ketamine-diazepam (KD) or propofol (P). Lactated Ringer's solution was administered by intravenous (IV) beginning 15 min before and continuing for 48 h after relief of urethral obstruction. Quality of recovery and time to standing were evaluated. The urethral catheter was maintained to measure urinary output. Hematocrit (Hct), total plasma protein (TPP), albumin, total protein (TP), blood urea nitrogen (BUN), creatinine, pH, bicarbonate (HCO3-), chloride, base excess, anion gap, sodium, potassium, and partial pressure of carbon dioxide in mixed venous blood (pvCO(2)) were measured before urethral obstruction, at start of fluid therapy (0 h), and at subsequent intervals. The quality of recovery and time to standing were respectively 4 and 75 min in the KD group and 5 and 16 min in the P group. The blood urea nitrogen values were increased at 0, 2, and 8 h in both groups. Serum creatinine increased at 0 and 2 h in cats administered KD and at 0, 2, and 8 h in cats receiving P, although the values were above the reference range in both groups until 8 h. Acidosis occurred for up to 2 h in both groups. Acid-base and biochemical stabilization were similar in cats anesthetized with propofol or with ketamine-diazepam. Cats that received propofol recovered much faster, but the ketamine-diazepam combination was shown to be more advantageous when treating uncooperative cats as it can be administered by intramuscular (IM) injection.

Grain size effect on the structural and dielectric properties of Pb0.85La0.15TiO3 ferroelectric ceramic compound

This paper presents a study of the influence of particle size on the structural and dielectric properties of Pb0.85La0.15TiO3 (PLT15) ferroelectric ceramic samples. The samples were prepared with average grain size of 1.69 +/- 0.08 mu m and 146 +/- 8 nm using, respectively, conventional and spark plasma sintering techniques. A decrease in the tetragonality degree as the crystallite size decreased was explained by an internal stress caused by the existence of a large amount of grain boundaries. The local structure exhibited no significant modification and the dielectric measurements showed a diffuse phase transition and a reduction in the permittivity magnitude at T-m as the average grain size decreased. The nanostructured ceramic sample prepared at a relatively lower temperature and sintering time presented a dielectric constant value of approximately 2000 at room temperature. (c) 2012 Elsevier Ltd and Techna Group S.r.l. All rights reserved.

Expression of soluble and functional full-length human matrix metalloproteinase-2 in Escherichia coli

Characterization of the matrix metalloproteinase-2 (MMP-2) substrates and understanding of its function remain difficult because up to date preparations containing minor amounts of other eukaryotic proteins that are co-purified with MMP-2 are still used. In this work, the expression of a soluble and functional full-length recombinant human MMP-2 (rhMMP-2) in the cytoplasm of Escherichia coli is reported, and the purification of this metalloproteinase is described. Culture of this bacterium at 18 degrees C culminated in maintenance of the soluble and functional rhMMP-2 in the soluble fraction of the E. coli lysate and its purification by affinity with gelatin-sepharose yielded approximately 0.12 mg/L of medium. Western Blotting and zymographic analysis revealed that the most abundant form was the 72-kDa MMP-2, but some gelatinolytic bands corresponding to proteins with lower molecular weight were also detected. The obtained rhMMP-2 was demonstrated to be functional in a gelatinolytic fluorimetric assay, suggesting that the purified rhMMP-2 was correctly folded. The method described here involves fewer steps, is less expensive, and is less prone to contamination with other proteinases and MMP inhibitors as compared to expression of rhMMP-2 in eukaryotic tissue culture. This protocol will facilitate the use of the full-length rhMMP-2 expressed in bacteria and will certainly help researchers to acquire new knowledge about the substrates and biological activities of this important proteinase. (C) 2011 Elsevier B.V. All rights reserved.

Structural and Luminescence Properties of Silica-Based Hybrids Containing New Silylated-Diketonato Europium(III) Complex

A new betadiketonate ligand displaying a trimethoxysilyl group as grafting function and a diketone moiety as complexing site (TTA-Si = 4,4,4-trifluoro-2-(3-trimethoxysilyl)propyl)-1-3-butanedione (C4H3S)COCH[(CH2)(3)Si(OCH3)(3)]COCF3) and its highly luminescent europium(III) complex [Eu(TTA-Si)(3)] have been synthesized and fully characterized. Luminescent silica-based hybrids have been prepared as well with this new complex grafted on the surface of dense silica nanoparticles (28 +/- 3 nm) or on mesoporous silica particles. The covalent bonding of Eu(TTA-Si)(3) inside the core of uniform silica nanoparticles (40 +/- 5 nm) was also achieved. Luminescence properties are discussed in relation to the europium chemical environment involved in each of the three hybrids. The general methodology proposed allowed high grafting ratios and overcame chelate release and tendency to agglomeration, and it could be applied to any silica matrix (in the core or at the surface, nanosized or not, dense or mesoporous) and therefore numerous applications such as luminescent markers and luminophors could be foreseen.

Structural and kinetic studies of Schistosoma mansoni adenylate kinases

The human parasite Schistosoma mansoni is totally dependent on the purine salvage pathway in order to supply large quantities of purine precursors for its energy and DNA biosynthetic needs. Adenylate kinase (ADK) is responsible for the conversion of AMP (produced by the adenosine kinase reaction) into ADP, which is subsequently converted into ATP by nucleoside diphosphate kinase (NDPK). ADK and NDPK are the most active enzymes of the pathway, probably reflecting an evolutionary adaptation due to the intense use of the branch in which they participate. However, notwithstanding their importance very little information has been accumulated found regarding these enzymes. In this work two adenylate kinases from S. mansoni were cloned and heterologously expressed in Escherichia coil. The purified products were utilized in activity assays, and displayed kinetic parameters similar to the corresponding human orthologous proteins. The cytosolic S. mansoni ADK was crystallized and its structure solved allowing us to detect a difference in the nucleotide binding site when compared with the human ortholog. (C) 2012 Elsevier B.V. All rights reserved.

Late evaluation of the relationship between morphological and functional renal changes and hypertension after non-operative treatment of high-grade renal injuries

Objective: To evaluate the anatomical and functional renal alterations and the association with post-traumatic arterial hypertension. Methods: The studied population included patients who sustained high grades renal injury (grades III to V) successfully non-operative management after staging by computed tomography over a 16-year period. Beyond the review of medical records, these patients were invited to the following protocol: clinical and laboratory evaluation, abdominal computed tomography, magnetic resonance angiography, DMSA renal scintigraphy, and ambulatory blood pressure monitoring. The hypertensive patients also were submitted to dynamic renal scintigraphy (Tc-99m EC), using captopril stimulation to verify renal vascular etiology. Results: Of the 31 patients, there were thirteen grade III, sixteen grade IV (nine lacerations, and seven vascular lesions), and two grade V injuries. All the patients were asymptomatic and an average follow up post-injury of 6.4 years. None had abnormal BUN or seric creatinine. The percentage of renal volume reduction correlates with the severity as defined by OIS. There was no evidence of renal artery stenosis in Magnetic Resonance angiography (MRA). DMSA scanning demonstrated a decline in percentage of total renal function corresponding to injury severity (42.2 +/- 5.5% for grade III, 35.3 +/- 12.8% for grade IV, 13.5 +/- 19.1 for grade V). Six patients (19.4%) had severe compromised function (< 30%). There was statistically significant difference in the decrease in renal function between parenchymal and vascular causes for grade IV injuries (p < 0.001). The 24-hour ambulatory blood pressure monitoring detected nine patients (29%) with post-traumatic hypertension. All the patients were male, mean 35.6 years, 77.8 % had a familial history of arterial hypertension, 66.7% had grade III renal injury, and average post-injury time was 7.8 years. Seven patients had negative captopril renography. Conclusions: Late results of renal function after conservative treatment of high grades renal injuries are favorable, except for patients with grades IV with vascular injuries and grade V renal injuries. Moreover, arterial hypertension does not correlate with the grade of renal injury or reduction of renal function.

A novel protein refolding protocol for the solubilization and purification of recombinant peptidoglycan-associated lipoprotein from Xylella fastidiosa overexpressed in Escherichia coil

Xylella fastidiosa is a Gram-negative xylem-limited plant pathogenic bacterium responsible for several economically important crop diseases. Here, we present a novel and efficient protein refolding protocol for the solubilization and purification of recombinant X. fastidiosa peptidoglycan-associated lipoprotein (XfPal). Pal is an outer membrane protein that plays important roles in maintaining the integrity of the cell envelope and in bacterial pathogenicity. Because Pal has a highly hydrophobic N-terminal domain, the heterologous expression studies necessary for structural and functional protein characterization are laborious once the recombinant protein is present in inclusion bodies. Our protocol based on the denaturation of the XfPal-enriched inclusion bodies with 8 M urea followed by buffer-exchange steps via dialysis proved effective for the solubilization and subsequent purification of XfPal, allowing us to obtain a large amount of relatively pure and folded protein. In addition, XfPal was biochemically and functionally characterized. The method for purification reported herein is valuable for further research on the three-dimensional structure and function of Pal and other outer membrane proteins and can contribute to a better understanding of the role of these proteins in bacterial pathogenicity, especially with regard to the plant pathogen X. fastidiosa. (C) 2012 Elsevier Inc. All rights reserved.

Metabolic and functional effects of beta-hydroxy-beta-methylbutyrate (HMB) supplementation in skeletal muscle

Beta-hydroxy-beta-methylbutyrate (HMB) is a metabolite derived from leucine. The anti-catabolic effect of HMB is well documented but its effect upon skeletal muscle strength and fatigue is still uncertain. In the present study, male Wistar rats were supplemented with HMB (320 mg/kg per day) for 4 weeks. Placebo group received saline solution only. Muscle strength (twitch and tetanic force) and resistance to acute muscle fatigue of the gastrocnemius muscle were evaluated by direct electrical stimulation of the sciatic nerve. The content of ATP and glycogen in red and white portions of gastrocnemius muscle were also evaluated. The effect of HMB on citrate synthase (CS) activity was also investigated. Muscle tetanic force was increased by HMB supplementation. No change was observed in time to peak of contraction and relaxation time. Resistance to acute muscle fatigue during intense contractile activity was also improved after HMB supplementation. Glycogen content was increased in both white (by fivefold) and red (by fourfold) portions of gastrocnemius muscle. HMB supplementation also increased the ATP content in red (by twofold) and white (1.2-fold) portions of gastrocnemius muscle. CS activity was increased by twofold in red portion of gastrocnemius muscle. These results support the proposition that HMB supplementation have marked change in oxidative metabolism improving muscle strength generation and performance during intense contractions.

Quantitative proteomic analysis and functional studies reveal that nucleophosmin is involved in cell death in glioblastoma cell line transfected with siRNA

Previously, we reported that nucleophosmin (NPM) was increased in glioblastoma multiforme (GBM). NPM is a phosphoprotein related to apoptosis, ribosome biogenesis, mitosis, and DNA repair, but details about its function remain unclear. We treated U87MG and A172 cells with small interference RNA (siRNA) and obtained a reduction of 80% in NPM1 expression. Knockdown at the protein level was evident after the 4th day and was maintained until the 7th day of transfection that was investigated by quantitative proteomic analysis using isobaric tags. The comparison of proteomic analysis of NPM1-siRNA against controls allowed the identification of 14 proteins, two proteins showed increase and 12 presented a reduction of expression levels. Gene ontology assigned most of the hypoexpressed proteins to apoptosis regulation, including GRP78. NPM1 silencing did not impair cell proliferation until the 7th day after transfection, but sensitized U87MG cells to temozolomide (TMZ), culminating with an increase in cell death and provoking at a later period a reduction of colony formation. In a large data set of GBM patients, both GRP78 and NPM1 genes were upregulated and presented a tendency to shorter overall survival time. In conclusion, NPM proved to participate in the apoptotic process, sensitizing TMZ-treated U87MG and A172 cells to cell death, and in association with upregulation of GRP78 may be helpful as a predictive factor of poor prognosis in GBM patients.