Thermoperiod affects the diurnal cycle of nitrate reductase expression and activity in pineapple plants by modulating the endogenous levels of cytokinins

Nitrate reductase (NR, EC 1.6.6.1) activity in higher plants is regulated by a variety of environmental factors and oscillates with a characteristic diurnal rhythm. In this study, we have demonstrated that the diurnal cycle of NR expression and activity in pineapple (Ananas comosus, cv. Smooth Cayenne) can be strongly modified by changes in the day/night temperature regime. Plants grown under constant temperature (28 degrees C light/dark) showed a marked increase in the shoot NR activity (NRA) during the first half of the light period, whereas under thermoperiodic conditions (28 degrees C light/15 degrees C dark) significant elevations in the NRA were detected only in the root tissues at night. Under both conditions, increases in NR transcript levels occurred synchronically about 4 h prior to the corresponding elevation of the NRA. Diurnal analysis of endogenous cytokinins indicated that transitory increases in the levels of zeatin, zeatin riboside and isopentenyladenine riboside coincided with the accumulation of NR transcripts and preceded the rise of NRA in the shoot during the day and in the root at night, suggesting these hormones as mediators of the temperature-induced modifications of the NR cycle. Moreover, these cytokinins also induced NRA in pineapple when applied exogenously. Altogether, these results provide evidence that thermoperiodism can modify the diurnal cycle of NR expression and activity in pineapple both temporally and spatially, possibly by modulating the day/night changes in the cytokinin levels. A potential relationship between the day/night NR cycle and the photosynthetic pathway performed by the pineapple plants (C(3) or CAM) is also discussed.

Human immature dental pulp stem cells share key characteristic features with limbal stem cells

Objectives: Limbal stem cells (LSC) are self-renewing, highly proliferative cells in vitro, which express a set of specific markers and in vivo have the capacity to reconstruct the entire corneal epithelium in cases of ocular surface injury. Currently, LSC transplantation is a commonly used procedure in patients with either uni- or bilateral total limbal stem cells deficiency (TLSCD). Although LSC transplantation holds great promise for patients, several problems need to be overcome. In order to find an alternative source of cells that can partially substitute LSC in cornea epithelium reconstruction, we aimed at investigating whether human immature dental pulp stem cells (hIDPSC) would present similar key characteristics as LSC and whether they could be used for corneal surface reconstruction in a rabbit TLSCD model. Materials: We used hIDPSC, which co-express mesenchymal and embryonic stem cell markers and present the capacity to differentiate into derivative cells of the three germinal layers. TLSCD was induced by chemical burn in one eye of rabbits. After 30 days, the opaque tissue formed was removed by superficial keratectomy. Experimental group received undifferentiated hIDPSC, while control group only received amniotic membrane (AM). Both groups were sacrificed after 3 months. Results and conclusions: We have demonstrated, using immunohistochemistry and reverse transcription-polymerase chain reaction, that hIDPSCs express markers in common with LSC, such as ABCG2, integrin beta 1, vimentin, p63, connexin 43 and cytokeratins 3/12. They were also capable of reconstructing the eye surface after induction of unilateral TLSCD in rabbits, as shown by morphological and immunohistochemical analysis using human-specific antibodies against limbal and corneal epithelium. Our data suggest that hIDPSCs share similar characteristics with LSC and might be used as a potential alternative source of cells for corneal reconstruction.

Nitric Oxide Mediates the Hormonal Control of Crassulacean Acid Metabolism Expression in Young Pineapple Plants

Genotypic, developmental, and environmental factors converge to determine the degree of Crassulacean acid metabolism (CAM) expression. To characterize the signaling events controlling CAM expression in young pineapple (Ananas comosus) plants, this photosynthetic pathway was modulated through manipulations in water availability. Rapid, intense, and completely reversible up-regulation in CAM expression was triggered by water deficit, as indicated by the rise in nocturnal malate accumulation and in the expression and activity of important CAM enzymes. During both up-and down-regulation of CAM, the degree of CAM expression was positively and negatively correlated with the endogenous levels of abscisic acid (ABA) and cytokinins, respectively. When exogenously applied, ABA stimulated and cytokinins repressed the expression of CAM. However, inhibition of water deficit-induced ABA accumulation did not block the up-regulation of CAM, suggesting that a parallel, non-ABA-dependent signaling route was also operating. Moreover, strong evidence revealed that nitric oxide (NO) may fulfill an important role during CAM signaling. Up-regulation of CAM was clearly observed in NO-treated plants, and a conspicuous temporal and spatial correlation was also evident between NO production and CAM expression. Removal of NO from the tissues either by adding NO scavenger or by inhibiting NO production significantly impaired ABA-induced up-regulation of CAM, indicating that NO likely acts as a key downstream component in the ABA-dependent signaling pathway. Finally, tungstate or glutamine inhibition of the NO-generating enzyme nitrate reductase completely blocked NO production during ABA-induced up-regulation of CAM, characterizing this enzyme as responsible for NO synthesis during CAM signaling in pineapple plants.

Saccharification of Brazilian sisal pulp: evaluating the impact of mercerization on non-hydrolyzed pulp and hydrolysis products

Cultivation of sisal, a plant with a short growth cycle, is highly productive in Brazil. This work is part of extensive research in which sisal is valued. In these studies, sisal fibers are used in the preparation of bio-based composites and in the derivatization of the pulp, including posterior preparation of films. This study aimed to examine the use of sisal pulp in the production of bioethanol, which can potentially be a high efficiency process because of the cellulose content of this fiber. A previous paper addressed the hydrolysis of sisal pulp using sulfuric acid as a catalyst. In the present study, the influence of the mercerization process on the acid hydrolysis of sisal pulp was evaluated. Mercerization was achieved in a 20% wt NaOH solution, and the cellulosic pulp was suspended and vigorously mixed for 1, 2 and 3 h, at 50 A degrees C. The previously characterized mercerized pulps were hydrolyzed (100 A degrees C, 30% H2SO4, v/v), and the results are compared with those obtained for unmercerized pulp (described in a companion paper). The starting sample was characterized by viscometry, alpha-cellulose content, crystallinity index and scanning electron microscopy. During the reactions, aliquots were withdrawn, and the liquor was analyzed by HPLC. The residual pulps (non-hydrolyzed) were also characterized by the techniques described for the initial sample. The results revealed that pretreatment decreases the polyoses content as well as causes a decrease of up to 23% in the crystallinity and up to 21% in the average molar mass of cellulose after 3 h of mercerization. The mercerization process proved to be very important to achieve the final target. Under the same reaction conditions (30% and 100 A degrees C, 6 h), the hydrolysis of mercerized pulp generated yields of up to 50% more glucose. The results of this paper will be compared with the results of subsequent studies obtained using other acids, and enzymes, as catalysts.

Antioxidant, anti-acetylcholinesterase and cytotoxic activities of ethanol extracts of peel, pulp and seeds of exotic Brazilian fruits Antioxidant, anti-acetylcholinesterase and cytotoxic activities in fruits

Ethanol extracts of powdered genipap (Genipa americana L), umbu (Spondia tuberosa A.) and siriguela (Spondia purpurea L) prepared from separate pulp, seeds and peel were investigated for their (i) antioxidant capacity, which was evaluated by various known methods; (ii) acetylcholinesterase (AChE) inhibitory activity; and (iii) cytotoxic effect on corneal epithelial cells of sheep. The highest values of total phenolic content were obtained with peel and seed extracts. Siriguela and umbu (seeds and peel) extracts displayed the highest antioxidant activities. Lipid peroxidation assays using mimetic biomembranes and mouse liver homogenates indicated that genipap pulp is a promising antioxidant. The investigation of phenols and organic acid contents revealed the presence of quercetin, citric and quinic acids, chlorogenic acid derivatives, among others, in several extracts, with the highest amount found in siriguela seeds. Genipap pulp and siriguela seed ethanol extracts presented an AChE inhibition zone similar to that of the positive control, carbachol. AChE inhibition assay with chlorogenic acid, one of the main constituents of siriguela seeds, revealed that this acid showed activity similar to that of the control physostigmine. These data suggest that these extracts are potentially important antioxidant supplements for the everyday human diet, pharmaceutical and cosmetic industries. (C) 2012 Elsevier Ltd. All rights reserved.

Bioactive compounds and phenolic-linked functionality of powdered tropical fruit residues

Tropical fruit residues consisting of seeds, peels and residual pulp generated as by-products of fruit processing industry were investigated for bioactive compounds, the in vitro antioxidant capacity as well as alpha-glucosidase and alpha-amylase inhibitory activities. Cyanidin, quercetin, ellagic acid (EA) and proanthocyanidins were found in acerola, jambolan, pitanga and caja-umbu residue powders. Acerola powder had the highest phenolic content (8839.33 mg catechin equivalents (CE)/100 g) and also high-ascorbic acid (AA) concentration (2748.03 mg/100 g), followed by jambolan and pitanga. The greatest 1,1-Diphenyl-2-picrylhydrazyl (DPPH) inhibition was observed for jambolan (436.76 mmol Trolox eq/g) followed by pitanga (206.68 mmol Trolox eq/g) and acerola (192.60 mmol Trolox eq/g), while acerola had the highest ferric reducing antioxidant power (FRAP) assay result (7.87 mmol Trolox eq/g). All fruit powders exhibited enzymatic inhibition against alpha-amylase (IC50 ranging from 3.40 to 49.5 mg CE/mL) and alpha-glucosidase (IC50 ranging from 1.15 to 2.37 mg CE/mL). Therefore, acerola, jambolan and pitanga dried residues are promising natural ingredients for food and nutraceutical manufacturers, due to their rich bioactive compound content.

Insecticidal Activity of the Granulosis Virus in Combination with Neem Products and Talc Powder Against the Potato Tuberworm Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae)

The potato tuberworm Phthorimaea operculella (Zeller) is an important agricultural pest that causes significant economic losses to potato growers worldwide. The addition of an effective method of biological control for the potato tuberworm is greatly needed, and is currently unavailable in Brazil. The granulosis virus (Baculoviridae) is a promising biological control agent to protect post-harvest potatoes and in storage from the potato tuberworm. However, the control measure must be economically feasible. Liquid suspensions of a granulosis virus applied alone or in mixture with two commercial neem oil-based products (DalNeem (TM) and NeemAzal (TM)), and a dry powder formulation of viral granules were evaluated for control of potato tuberworm larvae by treating potato tubers under laboratory conditions. High larval mortality (86.7%) was achieved when DalNeem and virus were applied together at 4 mg of azadirachtin/L and 10(4) occlusion bodies (OBs)/mL, respectively. This combination resulted in a parts per thousand yen50% efficacy in relation to their counterparts alone. Conversely, NeemAzal did not enhance virus effectiveness against larvae of the potato tuberworm. The talc-based virus formulation was used for dusting seed tubers at different concentrations and resulted in 100% larval mortality at 5 x 10(8) OBs/g. Formulated and unformulated virus provided 50% mortality at 166 OBs/g and at 5.0 x 10(5) OBs/mL, respectively. As a result, talc-based virus formulation had a better control efficiency on potato tuberworm than the aqueous virus suspension. The granulosis virus combined with DalNeem at low rates or formulated with talc powder is a viable option to control the potato tuberworm under storage conditions.

Integrated pulp and paper mill planning and scheduling

This article describes a real-world production planning and scheduling problem occurring at an integrated pulp and paper mill (P&P) which manufactures paper for cardboard out of produced pulp. During the cooking of wood chips in the digester, two by-products are produced: the pulp itself (virgin fibers) and the waste stream known as black liquor. The former is then mixed with recycled fibers and processed in a paper machine. Here, due to significant sequence-dependent setups in paper type changeovers, sizing and sequencing of lots have to be made simultaneously in order to efficiently use capacity. The latter is converted into electrical energy using a set of evaporators, recovery boilers and counter-pressure turbines. The planning challenge is then to synchronize the material flow as it moves through the pulp and paper mills, and energy plant, maximizing customer demand (as backlogging is allowed), and minimizing operation costs. Due to the intensive capital feature of P&P, the output of the digester must be maximized. As the production bottleneck is not fixed, to tackle this problem we propose a new model that integrates the critical production units associated to the pulp and paper mills, and energy plant for the first time. Simple stochastic mixed integer programming based local search heuristics are developed to obtain good feasible solutions for the problem. The benefits of integrating the three stages are discussed. The proposed approaches are tested on real-world data. Our work may help P&P companies to increase their competitiveness and reactiveness in dealing with demand pattern oscillations. (C) 2012 Elsevier Ltd. All rights reserved.

Advanced oxidation process H2O2/UV combined with anaerobic digestion to remove chlorinated organics from bleached kraft pulp mill wastewater

This study investigated the application of an advanced oxidation process combining hydrogen peroxide with ultraviolet radiation (H2O2/UV) to remove recalcitrant compounds from Kraft bleaching effluent. Anaerobic pre-treatment was performed to remove easily degraded organics using a horizontal-flow anaerobic immobilized biomass (HAIB) reactor. Bleaching plant effluent was treated in the HAIB reactor processed over 19 h of hydraulic retention time (HRT), reaching the expected removal efficiencies for COD (61 +/- 3%), TOC (69 +/- 9%), BOD5 (90 +/- 5%) and AOX (55 +/- 14%). However, the anaerobic treatment did not achieve acceptable removal of UV254 compounds. Furthermore, there was an increase of lignin, measured as total phenols. The H2O2/UV post-treatment provided a wide range of removal efficiencies depending on the dosage of hydrogen peroxide and UV irradiation: COD ranged from 0 to 11%, UV254 from 16 to 35%, lignin from 0 to 29% and AOX from 23 to 54%. All peroxide dosages applied in this work promoted an increase in the BOD5/COD ratio of the wastewater. The experiments demonstrate the technical feasibility of using H2O2/UV for post-treatment of bleaching effluents submitted to anaerobic pre-treatment.

Cytogenetic instability of dental pulp stem cell lines

Human adult stem cells (hASCs) offer a potentially renewable source of cell types that are easily isolated and rapidly expanded for use in regenerative medicine and cell therapies without the complicating ethical problems that are associated with embryonic stem cells. However, the eventual therapeutic use of hASCs requires that these cells and their derivatives maintain their genomic stability. There is currently a lack of systematic studies that are aimed at characterising aberrant chromosomal changes in cultured ASCs over time. However, the presence of mosaicism and accumulation of karyotypic abnormalities within cultured cell subpopulations have been reported. To investigate cytogenetic integrity of cultured human dental stem cell (hDSC) lines, we analysed four expanded hDSC cultures using classical G banding and fluorescent in situ hybridisation (FISH) with X chromosome specific probe. Our preliminary results revealed that about 70% of the cells exhibited karyotypic abnormalities including polyploidy, aneuploidy and ring chromosomes. The heterogeneous spectrum of abnormalities indicates a high frequency of chromosomal mutations that continuously arise upon extended culture. These findings emphasise the need for the careful analysis of the cytogenetic stability of cultured hDSCs before they can be used in clinical therapies.

Spin-glass-like behavior of uncompensated surface spins in NiO nanoparticulated powder

Nickel oxide nonoparticles successfully synthesized by a polymer percursor method are studied in this work. The analysis of X-ray powder diffraction data provides a mean crystallite size of 22 +/- 2 nm which is in a good agreement with the mean size estimated from transmission electron microscopy images. Whereas the magnetization (M) vs. magnetic field (H) curve obtained at 5 K is consistent with a ferromagnetic component which coexists with an antiferromagnetic component, the presence of two peaks in the zero-field-cooled trace suggests the occurrence of two blocking process. The broad maximum at high temperature was associated with the thermal relaxation of uncompensated spins at the particle core and the low temperature peak was assigned to the freeze of surface spins clusters. Static and dynamic magnetic results suggest that the correlations of surface spins clusters show a spin-glass-like below T-g = 7.3 +/- 0.1 K with critical exponents zv = 9.7 +/- 0.5 and beta = 0.7 +/- 0.1, which are consistent with typical reported for spin-glass systems. (C) 2012 Elsevier B.V. All rights reserved.

RNA Interference of Endochitinases in the Sugarcane Endophyte Trichoderma virens 223 Reduces Its Fitness as a Biocontrol Agent of Pineapple Disease

The sugarcane root endophyte Trichoderma virens 223 holds enormous potential as a sustainable alternative to chemical pesticides in the control of sugarcane diseases. Its efficacy as a biocontrol agent is thought to be associated with its production of chitinase enzymes, including N-acetyl-beta-D-glucosaminidases, chitobiosidases and endochitinases. We used targeted gene deletion and RNA-dependent gene silencing strategies to disrupt N-acetyl-beta-D-glucosaminidase and endochitinase activities of the fungus, and to determine their roles in the biocontrol of soil-borne plant pathogens. The loss of N-acetyl-beta-D-glucosaminidase activities was dispensable for biocontrol of the plurivorous damping-off pathogens Rhizoctonia solani and Sclerotinia sclerotiorum, and of the sugarcane pathogen Ceratocystis paradoxa, the causal agent of pineapple disease. Similarly, suppression of endochitinase activities had no effect on R. solani and S. sclerotiorum disease control, but had a pronounced effect on the ability of T. virens 223 to control pineapple disease. Our work demonstrates a critical requirement for T. virens 223 endochitinase activity in the biocontrol of C. paradoxa sugarcane disease, but not for general antagonism of other soil pathogens. This may reflect its lifestyle as a sugarcane root endophyte.

Effect of enzymatic treatment on the cross-flow microfiltration of acai pulp: Analysis of the fouling and recovery of phytochemicals

This study aimed to investigate the effects of pectinase enzyme treatment of acai pulp on cross-flow microfiltration (CFMF) performance and on phytochemical and functional characteristics of their compounds. Analyses of fouling mechanisms were carried out through resistance in series and blocking in law models. The enzymatic treatment was conducted using Ultrazym(R) AFPL (Novozymes A/S) at 500 mg kg(-1) of acai pulp for 30 min at 35 degrees C. Before microfiltrations, untreated and enzyme-treated acai pulps were previously diluted in distilled water (1:3; w/v). CFMFs were conducted using commercial alpha-alumina (alpha-Al2O3) ceramic membranes (Andritz AG, Austria) of 0.2 mu m and 0.8 mu m pore sizes, and 0.0047 m(2) of filtration area. The microfiltration unit was operated in batch mode for 120 min at 25 degrees C and the fluid-dynamic conditions were transmembrane pressure of Delta P = 100 kPa and cross-flow velocity of 3 m s(-1) in turbulent flow. The highest values of permeate flux and accumulated permeate volume were obtained using enzyme-treated pulp and 0.2 mu m pore size membranes with steady flux values exceeding 100 L h(-1) m(-2). For the 0.8 mu m pore size membrane, the estimated total resistance after the microfiltration of enzyme-treated acai pulp was 21% lower than the untreated pulp, and for the 0.2 mu m pore size membrane, it was 18%. Cake filtration was the dominant mechanism in the early stages of most of the CFMF processes. After approximately 20 min, however, intermediate pore blocking and complete pore blocking contributed to the overall fouling mechanisms. The reduction of the antioxidant capacity of the permeates obtained after microfiltration of the enzyme-treated pulp was higher (p < 0.01) than that obtained using untreated pulp. For total polyphenols, on the contrary, the permeates obtained after microfiltration of the enzyme-treated pulp showed a lower mean reduction (p < 0.01) than those from the untreated pulp. The results show that the enzymatic treatment had a positive effect on the CFMF process of acai pulp. (C) 2012 Elsevier Ltd. All rights reserved.

Sensitive and fast determination of Sudan dyes in chilli powder by partial-filling micellar electrokinetic chromatography-tandem mass spectrometry

A fast and sensitive method for the simultaneous determination of Sudan dyes (I, II, III, and IV) in food samples was developed for the first time using partial filling micellar electrokinectic chromatography-mass spectrometry (MEKC-MS). The use of MEKC was essential to achieve the separation of these neutral analytes, while the partial filling technique was necessary to avoid the contamination of the ion source with non-volatile micelles. MEKC separation and MS detection conditions were optimized in order to achieve a fast, efficient, and sensitive separation of the four dyes. Filling 25% of the capillary with an MEKC solution containing 40 mM ammonium bicarbonate, 25 mM SDS, and 32.5% (v/v) acetonitrile, a baseline separation of the four azo-dyes was obtained in 10 min. Tandem MS was investigated in order to improve the sensitivity and selectivity of the analysis. Limits of detection (LOD) values 5, 8, 15, and 29 times better were obtained for Sudan III, I, II, and IV, respectively, using partial filling MEKC-MS/MS instead of partial filling MEKC-MS. Under optimized conditions, LOD from 0.05 to 0.2 mu g/mL were obtained. The suitability of the developed method was demonstrated through the fast and sensitive determination of Sudan I, II, III, and IV in spiked chilli powder samples. This determination could not be achieved by MEKC-UV due to the existence of several interfering compounds from the matrix.

Influence of pulp condition on the accuracy of an electronic foramen locator in posterior teeth: an in vivo study

The aim of this study was to assess, in vivo, the accuracy of the NovApex (R) electronic foramen locator in determining working length (WL) in vital and necrotic posterior teeth. The NovApex (R) was used in 144 canals: 35 teeth with vital pulps (68 canals) and 42 teeth with necrotic pulps (76 canals). WL was measured with the NovApex (R) locator and confirmed using the radiographic method. Differences between electronic and radiographic measurements ranging between 0.0 and 0.4 millimeters were classified as acceptable; differences equal to or greater than 0.5 millimeter were considered unacceptable. Pearson's chi-square test was used to assess the influence of pulp condition on the accuracy of NovApex (R) (alpha = 0.05). Regardless of pulp condition, differences between electronic and radiographic WL measurements were acceptable in 73.61% of the canals. No statistically significant differences in accuracy were observed when comparing vital and necrotic canals (p > 0.05). There were 38 unacceptable measurements. In none of these cases was the file tip located beyond the radiographic apex; in 32, it was located short of the NovApex (R) measurement. Pulp condition had no significant effect on the accuracy of NovApex (R).