Supercritical CO(2) extraction of carotenoids from pitanga fruits (Eugenia uniflora L.)

Supercritical carbon dioxide (SC-CO(2)) extraction was employed to extract carotenoids from the freeze-dried pulp of pitanga fruits (Eugenia uniflora L.), an exotic fruit, rich in carotenoids and still little explored commercially. The SC-CO(2) extraction was carried out at two temperatures, 40 and 60 degrees C, and seven pressures, 100, 150, 200, 250, 300, 350 and 400 bar. The carotenoids were determined by high-performance liquid chromatography connected to photodiode array and mass spectrometry detectors. Lycopene, rubixanthin and P-cryptoxanthin were the main carotenoids present in the freeze-dried pitanga pulp, whereas beta-cryptoxanthin concentration was negligible in the SC-CO(2) extracts, for all the investigated state conditions. The maximum recovery of carotenoids was obtained at 60 degrees C and 250 bar, extracting 55% of the total carotenoid content, 74% of the rubixanthin and 78% of the lycopene from the pulp. Under these state conditions, the total carotenoid concentration in the extract was 5474 mu g/g, represented by 66% lycopene and 32% rubixanthin. The experimental state conditions produced different SC-CO(2) extracts with respect to the extraction yield and concentration of different carotenoids, indicating that the supercritical carbon dioxide was selective in the extraction of the pitanga carotenoids as a function of temperature and pressure. (C) 2008 Elsevier B.V. All rights reserved.

Underutilized Annona Species from the Brazilian Cerrado and Amazon Rainforest: A Study on Fatty Acids Profile and Yield of Seed Oils

Annona (Annonaceae) is an important source of fruits in the Brazilian Cerrado and the Amazon Rainforest. Some Annona species are widely commercialized as fresh fruit or as frozen pulp. Seeds are accustomedly discarded. Our main goal was to analyze fatty acids profile from seeds of A. crassiflora and A. coriacea from Cerrado, A. montana from Amazon Forest, and three cultivars (A. cherimola cv. Madeira, A. cherimola x A. squamosa cv. Pink`s Mammonth and A. cherimola x A. squamosa cv. Gefner). The total oil yield ranged between 20 and 42% by weight of dry mass. The A cherimola x A. squamosa cv. Gefner has significantly higher total lipid yield than all other samples. 100 g of fruit of this species present 6-8 g of seeds. Considering the fruit production of Chile (over 221 ton of fruits/year), more than 1300 ton of seed/year could be obtained, which could provide at least 200 ton of seed oil. Oleic acid was predominant for most samples, but for A. montana linoleic acid was the most abundant FA. Phenotypic variation on FAME profile was observed. These new data are an urgent requirement for supporting conservation programs, mainly for Cerrado areas in Brazil.

Human Multipotent Mesenchymal Stromal Cells from Distinct Sources Show Different In Vivo Potential to Differentiate into Muscle Cells When Injected in Dystrophic Mice

Limb-girdle muscular dystrophies are a heterogeneous group of disorders characterized by progressive degeneration of skeletal muscle caused by the absence or deficiency of muscle proteins. The murine model of Limb-Girdle Muscular Dystrophy 2B, the SJL mice, carries a deletion in the dysferlin gene. Functionally, this mouse model shows discrete muscle weakness, starting at the age of 4-6 weeks. The possibility to restore the expression of the defective protein and improve muscular performance by cell therapy is a promising approach for the future treatment of progressive muscular dystrophies (PMD). We and others have recently shown that human adipose multipotent mesenchymal stromal cells (hASCs) can differentiate into skeletal muscle when in contact with dystrophic muscle cells in vitro and in vivo. Umbilical cord tissue and adipose tissue are known rich sources of multipotent mesenchymal stromal cells (MSCs), widely used for cell-based therapy studies. The main objective of the present study is to evaluate if MSCs from these two different sources have the same potential to reach and differentiate in muscle cells in vivo or if this capability is influenced by the niche from where they were obtained. In order to address this question we injected human derived umbilical cord tissue MSCs (hUCT MSCs) into the caudal vein of SJL mice with the same protocol previously used for hASCs; we evaluated the ability of these cells to engraft into recipient dystrophic muscle after systemic delivery, to express human muscle proteins in the dystrophic host and their effect in functional performance. These results are of great interest for future therapeutic application.

Human Stem Cell Cultures from Cleft Lip/Palate Patients Show Enrichment of Transcripts Involved in Extracellular Matrix Modeling By Comparison to Controls

Nonsyndromic cleft lip and palate (NSCL/P) is a complex disease resulting from failure of fusion of facial primordia, a complex developmental process that includes the epithelial-mesenchymal transition (EMT). Detection of differential gene transcription between NSCL/P patients and control individuals offers an interesting alternative for investigating pathways involved in disease manifestation. Here we compared the transcriptome of 6 dental pulp stem cell (DPSC) cultures from NSCL/P patients and 6 controls. Eighty-seven differentially expressed genes (DEGs) were identified. The most significant putative gene network comprised 13 out of 87 DEGs of which 8 encode extracellular proteins: ACAN, COL4A1, COL4A2, GDF15, IGF2, MMP1, MMP3 and PDGFa. Through clustering analyses we also observed that MMP3, ACAN, COL4A1 and COL4A2 exhibit co-regulated expression. Interestingly, it is known that MMP3 cleavages a wide range of extracellular proteins, including the collagens IV, V, IX, X, proteoglycans, fibronectin and laminin. It is also capable of activating other MMPs. Moreover, MMP3 had previously been associated with NSCL/P. The same general pattern was observed in a further sample, confirming involvement of synchronized gene expression patterns which differed between NSCL/P patients and controls. These results show the robustness of our methodology for the detection of differentially expressed genes using the RankProd method. In conclusion, DPSCs from NSCL/P patients exhibit gene expression signatures involving genes associated with mechanisms of extracellular matrix modeling and palate EMT processes which differ from those observed in controls. This comparative approach should lead to a more rapid identification of gene networks predisposing to this complex malformation syndrome than conventional gene mapping technologies.

CCR2 Deficiency Results in Increased Osteolysis in Experimental Periapical Lesions in Mice

Introduction: Periapical lesions are chronic inflammatory disorders of periradicular tissues caused by etiologic agents of endodontic origin. The inflammatory chemokines are thought to be involved in the latter observed osteolysis. With a murine model of experimental periapical lesion, the objective of this study was to evaluate the role of the chemokine receptor CCR2 in the lesion progression, osteoclast differentiation and activation, and expression of inflammatory osteolysis-related mediators. Methods: For lesion induction, right mandibular first molars were opened surgically with a (1)/(4) carbine bur, and 4 bacterial strains were inoculated in the exposed dental pulp; left mandibular first molars were used as controls. Animals were killed at 3, 7, 14, and 21 days after surgeries to evaluate the kinetics of lesion development. Results: CCR2 KO mice showed wider lesions than WT mice. CCR2 KO mice also expressed higher levels of the osteoclastogenic and osteolytic factors, receptor activator of nuclear factor kappa B ligand (RANKL) and cathepsin K, of the proinflammatory cytokine tumor necrosis factor alpha, and of the neutrophil migration related chemokine, KC. Conclusions: These results suggest that CCR2 is important in host protection to periapical osteolysis. (J Endod 2010;36:244-250)

Comparative Histopathological Analysis of Human Pulps after Class I Cavity Preparation with a High-Speed Air-Turbine Handpiece or Er:YAG Laser

The purpose of this study was to comparatively evaluate the response of human pulps after cavity preparation with different devices. Deep class I cavities were prepared in sound mandibular premolars using either a high-speed air-turbine handpiece (Group 1) or an Er: YAG laser (Group 2). Following total acid etching and the application of an adhesive system, all cavities were restored with composite resin. Fifteen days after the clinical procedure, the teeth were extracted and processed for analysis under optical microscopy. In Group 1 in which the average for the remaining dentin thickness (RDT) between the cavity floor and the coronal pulp was 909.5 mu m, a discrete inflammatory response occurred in only one specimen with an RDT of 214 mu m. However, tissue disorganization occurred in most specimens. In Group 2 (average RDT = 935.2 mu m), the discrete inflammatory pulp response was observed in only one specimen (average RDT = 413 mu m). It may be concluded that the high-speed air-turbine handpiece caused greater structural alterations in the pulp, although without inducing inflammatory processes.

Bismuth germanate films prepared by Pechini method

Bismuth germanate films were prepared by dip coating and spin coating techniques and the dependence of the luminescent properties of the samples on the resin viscosity and deposition technique was investigated. The resin used for the preparation of the films was obtained via Pechini method, employing the precursors Bi(2)O(3) and GeO(2). Citric acid and ethylene glycol were used as chelating and cross-linking agents, respectively. Results from X-ray diffraction and Raman spectroscopy indicated that the films sintered at 700 degrees C for 10 h presented the single crystalline phase Bi(4)Ge(3)O(12). SEM images of the films have shown that homogeneous flat films can be produced by the two techniques investigated. All the samples presented the typical Bi(4)Ge(3)O(12) emission band centred at 505 nm. Films with 3.1 mu m average thickness presented 80% of the luminescence intensity registered for the single crystal at the maximum wavelength. Published by Elsevier B.V.

Increased Viability of Odontoblast-Like Cells Subjected to Low-Level Laser Irradiation

Studies have shown that the increase of cell metabolism depends on the low level laser therapy (LLLT) parameters used to irradiate the cells. However, the optimal laser dose to up-regulate pulp cell activity remains unknown. Consequently, the aim of this study was to evaluate the metabolic response of odontoblast-like cells (MDPC-23) exposed to different LLLT doses. Cells at 20000 cells/cm(2) were seeded in 24-well plates using plain culture medium (DMEM) and were incubated in a humidified incubator with 5% CO(2) at 37 degrees C. After 24 h, the culture medium was replaced by fresh DMEM supplemented with 5% (stress by nutritional deficit) or 10% fetal bovine serum (FBS). The cells were exposed to different laser doses from a near infrared diode laser prototype designed to provide a uniform irradiation of the wells. The experimental groups were: G1: 1.5 J/cm(2) + 5% FBS; G2: 1.5 J/cm(2) + 10% FBS; G3: 5 J/cm(2) + 5% FBS; G4: 5 J/cm(2) + 10% FBS; G5: 19 J/cm(2) + 5% FBS; G6: 19 J/cm(2) + 10% FBS. LLLT was performed in 3 consecutive irradiation cycles with a 24-hour interval. Non-irradiated cells cultured in DMEM supplemented with either 5 or 10% FBS served as control groups. The analysis of the metabolic response was performed by the MTT assay 3 h after the last irradiation. G1 presented an increase in SDH enzyme activity and differed significantly (Mann-Whitney test, p < 0.05) from the other groups. Analysis by scanning electron microscopy showed normal cell morphology in all groups. Under the tested conditions, LLLT stimulated the metabolic activity of MDPC-23 cultured in DMEM supplemented with 5% FBS and exposed to a laser dose of 1.5 J/cm(2). These findings are relevant for further studies on the action of near infrared lasers on cells with odontoblast phenotype.

Total phenolic content and free radical scavenging activities of methanolic extract powders of tropical fruit residues

Methanolic extract powders of acerola, passion fruit and pineapple industrial residues, including pulp, seeds and peel, altogether (except for acerola) devoid of seeds, were screened for antioxidant capacity. The total phenolic contents (TPCs) of the extract powders were compared with their radical-scavenging activities (RSA) against both DPPH(center dot) and superoxide anion (O(2)(center dot-)) radicals, and their protective effect against liposome peroxidation, triggered by peroxyl radical. Lipid peroxidation was followed by the fluorescence decay of the probe, 4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3-undecanoic acid (C(11)-BODIPY(581/591)). The TPCs of acerola, passion fruit and pineapple extract powders were (94.6 +/- 7.4); (41.2 +/- 4.2) and (9.1 +/- 1.3) mg of gallic acid equivalents g(-1) of dry extract, respectively. Acerola showed the best RSA-DPPH(center dot) scores, whereas passion fruit was more protective on the RSA-O(2)(center dot-) system. Together with the protective effects against lipid peroxidation (rate of BODIPY decay) which, were similar for acerola and passion fruit extracts, these data suggest that the methanolic extracts of acerola and passion fruit residues may be useful as antioxidant supplements, particularly the acerola extract, due to its high phenolic content. (C) 2008 Elsevier Ltd. All rights reserved

cis-4-decenoic acid provokes mitochondrial bioenergetic dysfunction in rat brain

Aims: In the present work we investigated the in vitro effect of cis-4-decenoic acid, the pathognomonic metabolite of medium-chain acyl-CoA dehydrogenase deficiency, on various parameters of bioenergetic homeostasis in rat brain mitochondria. Main methods: Respiratory parameters determined by oxygen consumption were evaluated, as well as membrane potential, NAD(P)H content, swelling and cytochrome c release in mitochondrial preparations from rat brain, using glutamate plus malate or succinate as substrates. The activities of citric acid cycle enzymes were also assessed. Key findings: cis-4-decenoic acid markedly increased state 4 respiration, whereas state 3 respiration and the respiratory control ratio were decreased. The ADP/O ratio, the mitochondrial membrane potential, the matrix NAD(P)H levels and aconitase activity were also diminished by cis-4-decenoic acid. These data indicate that this fatty acid acts as an uncoupler of oxidative phosphorylation and as a metabolic inhibitor. cis-4-decenoic acid also provoked a marked mitochondrial swelling when either KCl or sucrose was used in the incubation medium and also induced cytochrome c release from mitochondria, suggesting a non-selective permeabilization of the inner mitochondria! membrane. Significance: It is therefore presumed that impairment of mitochondrial homeostasis provoked by cis-4-decenoic acid may be involved in the brain dysfunction observed in medium-chain acyl-CoA dehydrogenase deficient patients. (C) 2010 Elsevier Inc. All rights reserved.

Water-soluble Cu, Fe, Mn and Zn species in nuts and seeds

An approach was developed to estimate molecular weight distribution of water-soluble Cu, Fe, Mn and Zn species in Brazil nut, cupuassu seed and coconut pulp by size exclusion chromatography (SEC) coupled on-line to ultra-violet (UV) and off-line to graphite furnace atomic absorption spectrometry (GF-AAS) detectors and matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF-MS). SEC-UV analytical signals showed the prevalence of high molecular weight (HMW) species (79-1.7 kDa for Brazil nut, 50-1.7 kDa for coconut pulp, and 34-1.7 kDa for cupuassu seeds). The Brazil nut SEC-UV, GF-AAS and MALDI-TOF mass spectra gave confirmation of the association of the elements with water-soluble compounds. The elemental profiles were associated with fractions of compounds of molecular weight 1.2-16 kDa for Brazil nut, 1.7-13 kDa for coconut pulp, and 1.2-7.6 kDa for cupuassu seeds. (C) 2009 Elsevier Inc. All rights reserved.

Flow injection analysis of ethyl xanthate by gas diffusion and UV detection as CS(2) for process monitoring of sulfide ore flotation

A sensitive and robust analytical method for spectrophotometric determination of ethyl xanthate, CH(3)CH(2)OCS(2)(-) at trace concentrations in pulp solutions from froth flotation process is proposed. The analytical method is based on the decomposition of ethyl xanthate. EtX(-), with 2.0 mol L(-1) HCl generating ethanol and carbon disulfide. CS(2). A gas diffusion cell assures that only the volatile compounds diffuse through a PTFE membrane towards an acceptor stream of deionized water, thus avoiding the interferences of non-volatile compounds and suspended particles. The CS(2) is selectively detected by UV absorbance at 206 nm (epsilon = 65,000 L mol(-1) cm(-1)). The measured absorbance is directly proportional to EtX(-) concentration present in the sample solutions. The Beer`s law is obeyed in a 1 x 10(-6) to 2 x 10(-4) mol L(-1) concentration range of ethyl xanthate in the pulp with an excellent correlation coefficient (r = 0.999) and a detection limit of 3.1 x 10(-7) mol L(-1), corresponding to 38 mu g L. At flow rates of 200 mu L min(-1) of the donor stream and 100 mu L min(-1) of the acceptor channel a sampling rate of 15 injections per hour could be achieved with RSD < 2.3% (n = 10, 300 mu L injections of 1 x 10(-5) mol L(-1) EtX(-)). Two practical applications demonstrate the versatility of the FIA method: (i) evaluation the free EtX(-) concentration during a laboratory study of the EtX(-) adsorption capacity on pulverized sulfide ore (pyrite) and (ii) monitoring of EtX(-) at different stages (from starting load to washing effluents) of a flotation pilot plant processing a Cu-Zn sulfide ore. (C) 2010 Elsevier By. All rights reserved.

Elemental analysis of nuts and seeds by axially viewed ICP OES

A method for the multi-elemental determination of metals (Al, Ba, Ca, Cu, Fe, K, Mg, Mn, Sr and Zn), metalloids (B and Si), and non-metals (Cl, P and 5) in the babassu nut and mesocarp, sapucaia nut, coconut pulp, cupuassu pulp and seed, and cashew nut by axially viewed inductively coupled plasma optical emission spectrometry is presented. A diluted oxidant mixture (2 ml HNO(3) + 1 ml H(2)O(2) + 3 ml H(2)O) was used to achieve the complete decomposition of the organic matrix in a closed-vessel microwave oven. The accuracy of the entire proposed method was confirmed by standard reference material analysis (peach leaves-NIST SRM1547). The certified values showed a good agreement at a 95% confidence limit (Student`s t-test). The average RSD for repeatability of calibration solutions measurements were in the range of 1.1-6.7%. Limits of quantification (LOQ = 10 x LOD) were in the level of 0.00072-0.0532 mg/l. The macro and micronutrient ranges in the different nuts and seeds did not exceed the dietary reference intake (DRI), except for Mn in the babassu nut. (C) 2010 Published by Elsevier Ltd.

The corrosion of austenitic 304 stainless steel in biodiesel

Fuel distribution uses 304 stainless steel containers for the storage of biofuels, however there are few reports in the literature about the corrosive aspects this. steel in biodiesel. The objective of this research is to study the corrosive behavior of 304 austenitic stainless steel in the presence of biodiesel, unwashed and washed, with aqueous solutions of citric, oxalic, acetic and ascorbic acids 0,01 mol L(-1), and compare with results obtained for the copper (ASTM D130). The employedtechniques were: atomic absorption spectrometry (AAS) and optical microscopy (OM). The results of EA A showed a low rate of corrosion for the stainless steel, the alloys elements studied were Cr, Ni and Fe, the highest rate was observed for the chrome, 1.78 ppm / day in biodiesel with or without washing. The OM of the 304 steel, when compared with that of copper has a low corrosion rate in the 304 steel/biodiesel system. Not with standing, this demonstrates that not only the 304 steel, but also the copper corrodes in biodiesel

Hydrogen peroxide and supercritical carbon dioxide: a new bleaching stage for Eucalyptus kraft-O(2) pulps

Kraft pulp is currently bleached largely by the elemental chlorine free (ECF) technology with oxygen, chlorine dioxide, and hydrogen as active agents. This technology brought about significant environmental improvements in relation to standard processes based on chlorine gas and hypochlorite, but there is still need for further improvements. This study presents a novel environmentally friendly bleaching stage - the so-called `hydrogen peroxide in supercritical carbon dioxide`, P((SC-CO2)) - that can be adapted to current ECF bleaching processes, with preference in cases where hydrogen peroxide is already used. In this study, the P((SC-CO2)) stage was evaluated as a replacement to the last peroxide stage of the D(EP)DP bleaching sequence and to the first peroxide stage of the D(EP)DP sequence, for an oxygen delignified eucalypt kraft-O(2) pulp. The P((SC-CO2)) stage was run with 0.5% hydrogen peroxide, at 15% consistency, 70 degrees C, and 73 bar. The reaction time was 30 min. The performances of regular P stages and the new P((SC-CO2)) stage were compared. Promising results were observed with the DEP((SC-CO2))DP sequence; the P((SC-CO2)) decreased kappa number from 2.7 to 2.1, and the hexenuronic acid groups from 17.0 to 12.4 mmol kg(-1). The P((SC-CO2)) stage showed poor performance when applied in the D(EP)DP((SC-CO2)) sequence. It is concluded that the process presents potential but requires further optimization to improve selectivity and efficiency.