Expression of genes from the lignin synthesis pathway in guineagrass genotypes differing in cell-wall digestibility

Rapid decline in cell-wall digestibility hinders efficient use of warm-season grasses. The objective of this study was to identify genes whose expressions are related to the slope of decline in cell-wall digestibility. Eleven guineagrass genotypes were harvested at three ages and classified according to fibre digestibility. Extreme genotypes were separated into groups with either FAST or SLOW decline in fibre digestibility. Expression of transcripts from six genes from the lignin synthesis pathway was quantified by real-time PCR. Fast decline in fibre digestibility was associated with higher DM yield after 90 d of regrowth. Apart from lower fibre digestibility and higher lignin content for the FAST group, there were no other differences between the two groups for the chemical composition of stems and leaves. Maturity affected differently the expression of two of the six genes, cinnamate 4-hydroxylase and caffeoyl-CoA O-methyltransferase (C4H and CCoAOMT). Genotypes with fast decline in fibre digestibility had greater increase in the expression of C4H and CCoAOMT from 30 to 60 d of regrowth, than genotypes with slower decline. Expression of C4H and CCoAOMT appears to be related to the decline in cell-wall digestibility with advance in maturity of guineagrass.

Carboxymethyl lignin as stabilizing agent in aqueous ceramic suspensions

Identifying new uses for residues of industries that process large quantities of biomass, as in bioethanol production, is essential for a sustainable development with reduced impact on the environment, which is the reason why many efforts have been devoted to find noble uses for lignins. in this study, a lignin obtained from sugarcane bagasse in a bioethanol producing plant was carboxymethylated to yield the water-soluble carboxymethyl lignin (CML), which was then used as stabilizing agent in aqueous alumina (Al2O3) suspensions. CML had a degree of substitution 0.46 +/- 0.01, in relation to the C9 unit of lignin, and behaved as a polyelectrolyte in a large pH range owing to the dissociation of carboxylic groups. The action of CML as stabilizing agent of alumina aqueous suspensions was investigated using viscometry, zeta potential, and photon correlation spectroscopy (PCS) measurements, mainly as a function of pH and time. Overall, the results showed that CML had a good performance as a deflocculating agent, because it led to dispersions with low viscosity and small change in particle size as a function of time. The positive effect from the addition of CML was confirmed in the morphological features of the material obtained from the alumina suspensions after elimination of water, as indicated by scanning electron microscopy. The stabilization of alumina suspensions afforded by CML opens the way for similar applications of modified lignins, whose electrical and structural properties may be tuned for specific uses in various industries, including the ceramic industry. (C) 2011 Elsevier B.V. All rights reserved.

Direct assembly of a metallodendrimer encompassing seven triruthenium clusters units

A general strategy for the assembly of dendrimeric metallo-cluster species based on tritopic trinuclear ruthenium acetate complexes is demonstrated. First, a central core consisting of a [Ru3O(CH3COO)(6)(TPEB)(3)]PF6 complex (G0), where TPEB is the tripodal 1,3,5-tri-4-pyridyl-1,2-ethenylbenzene ligand, was synthesized and then reacted with the end-capping complex [Ru3O(CH3COO)(6)(py)(2)(MeOH)]PF6, thus composing the first generation shell of a dendrimer encompassing twenty-one ruthenium ions (G1). The core and dendrimeric complexes were characterized by elemental analysis, UV-Vis, H-1 NMR, ESI-MS spectrometry and Differential pulse voltammetry. All results were consistent with the structure of that multinuclear cationic dendrimeric species. The isotopologic profile of daughter fragments and the strength of the metal-ligand bonds were carefully investigated providing the fragmentation pathway for the metallo-dendrimer upon ESI-MS dissociation conditions. (C) 2012 Elsevier B.V. All rights reserved.

Structural features of lignin obtained at different alkaline oxidation conditions from sugarcane bagasse

Lignin is a macromolecule frequently obtained as residue during technological processing of biomass. Modifications in chemical structure of lignin generate valuable products, some with particular and unique characteristics. One of the available methods for modification of industrial lignin is oxidation by hydrogen peroxide. In this work, we conducted systematic studies of the oxidation process that were carried out at various pHs and oxidizing agent concentrations. Biophysical, biochemical, structural properties of the oxidized lignin were analyzed by UV spectrophotometry, Fourier transform infrared spectroscopy, scanning electron microscopy and small angle X-ray scattering. Our results reveal that lignin oxidized with 9.1% H(2)O(2) (m/v) at pH 13.3 has the highest fragmentation, oxidation degree and stability. Although this processing condition might be considered quite severe, we have concluded that the stability of the obtained oxidized lignin was greatly increased. Therefore, the identified processing conditions of oxidation may be of practical interest for industrial applications. (C) 2011 Elsevier B.V. All rights reserved.

The viewpoint of health professionals on the role of basic units in the healthcare network of the Brazilian Unified Health System

In the process of creation of the Unified Health System (SUS) as a universal policy seeking to ensure comprehensive care, unscheduled assistance in primary healthcare units (UBS) is an unresolved challenge. The scope of this paper is to analyze the viewpoint of health professionals on the role of primary healthcare units in meeting this demand. It is a transversal study of qualitative data obtained through questionnaires and interviews with 106 medical practitioners from 6 emergency medical services and 190 professionals from 30 units. They explained why people seek emergency care for occurrences pertaining to primary care. The content analysis technique with thematic categories was used for data analysis. Lack of resources and problems with primary health unit work processes (50.8%) were the reasons most frequently cited by emergency care physicians to explain this inadequate demand. Only 33.3% of the health unit professionals agreed that these occurrences should be attended in the primary healthcare services. The limited viewpoint of the role of health services on the unscheduled care, particularly among primary care professionals, possibly leads to restrictive practices for access by the population.

RELATIONSHIP AMONG EXTRACTIVES, LIGNIN AND HOLOCELLULOSE CONTENTS WITH PERFORMANCE INDEX OF SEVEN WOOD SPECIES USED FOR BOWS OF STRING INSTRUMENTS

In this paper we investigate the influence of extractives, lignin and holocellulose contents on performance index (PI) of seven woods used or tested for violin bows. Woods with higher values of this index (PI = root MOE/rho, where MOE is modulus of elasticity and rho is density) have a higher bending stiffness at a given mass, which can be related to bow wood quality. Extractive content was negatively correlated with PI in Caesalpinia echinata, Hanclroanthus sp. and Astronium lecointei. In C. echinata holocellulose was positively correlated with PI. These results need to be further explored with more samples and by testing additional wood properties. Although the chemical constituents could provide an indication of quality, it is not possible to establish appropriate woods for bows solely by examining their chemical constituents.

Mammography-based screening program: preliminary results from a first 2-year round in a Brazilian region using mobile and fixed units

Background: Breast cancer is the most frequently diagnosed cancer and the leading cause of cancer deaths among women worldwide. The use of mobile mammography units to offer screening to women living in remote areas is a rational strategy to increase the number of women examined. This study aimed to evaluate results from the first 2 years of a government-organized mammography screening program implemented with a mobile unit (MU) and a fixed unit (FU) in a rural county in Brazil. The program offered breast cancer screening to women living in Barretos and the surrounding area. Methods: Based on epidemiologic data, 54 238 women, aged 40 to 69 years, were eligible for breast cancer screening. The study included women examined from April 1, 2003 to March 31, 2005. The chi-square test and Bonferroni correction analyses were used to evaluate the frequencies of tumors and the importance of clinical parameters and tumor characteristics. Significance was set at p < 0.05. Results: Overall, 17 964 women underwent mammography. This represented 33.1% of eligible women in the area. A mean of 18.6 and 26.3 women per day were examined in the FU and MU, respectively. Seventy six patients were diagnosed with breast cancer (41 (54%) in the MU). This represented 4.2 cases of breast cancer per 1000 examinations. The number of cancers detected was significantly higher in women aged 60 to 69 years than in those aged 50 to 59 years (p < 0.001) or 40 to 49 years (p < 0.001). No difference was observed between women aged 40 to 49 years and those aged 50 to 59 years (p = 0.164). The proportion of tumors in the early (EC 0 and EC I) and advanced (CS III and CS IV) stages of development were 43.4% and 15.8%, respectively. Conclusions: Preliminary results indicate that this mammography screening program is feasible for implementation in a rural Brazilian territory and favor program continuation.

Toward the Optimization of an e-Tongue System Using Information Visualization: A Case Study with Perylene Tetracarboxylic Derivative Films in the Sensing Units

The wide variety of molecular architectures used in sensors and biosensors and the large amount of data generated with some principles of detection have motivated the use of computational methods, such as information visualization techniques, not only to handle the data but also to optimize sensing performance. In this study, we combine projection techniques with micro-Raman scattering and atomic force microscopy (AFM) to address critical issues related to practical applications of electronic tongues (e-tongues) based on impedance spectroscopy. Experimentally, we used sensing units made with thin films of a perylene derivative (AzoPTCD acronym), coating Pt interdigitated electrodes, to detect CuCl(2) (Cu(2+)), methylene blue (MB), and saccharose in aqueous solutions, which were selected due to their distinct molecular sizes and ionic character in solution. The AzoPTCD films were deposited from monolayers to 120 nm via Langmuir-Blodgett (LB) and physical vapor deposition (PVD) techniques. Because the main aspects investigated were how the interdigitated electrodes are coated by thin films (architecture on e-tongue) and the film thickness, we decided to employ the same material for all sensing units. The capacitance data were projected into a 2D plot using the force scheme method, from which we could infer that at low analyte concentrations the electrical response of the units was determined by the film thickness. Concentrations at 10 mu M or higher could be distinguished with thinner films tens of nanometers at most-which could withstand the impedance measurements, and without causing significant changes in the Raman signal for the AzoPTCD film-forming molecules. The sensitivity to the analytes appears to be related to adsorption on the film surface, as inferred from Raman spectroscopy data using MB as analyte and from the multidimensional projections. The analysis of the results presented may serve as a new route to select materials and molecular architectures for novel sensors and biosensors, in addition to suggesting ways to unravel the mechanisms behind the high sensitivity obtained in various sensors.

Shape resonance spectra of lignin subunits

We report integral cross sections for elastic electron scattering by the lignin subunits phenol, guaiacol, and p-coumaryl alcohol. Our calculations employed the Schwinger multichannel method with pseudopotentials and indicate three to four pi* shape resonances for each of these systems, suggesting that low-energy electrons could efficiently transfer energy into the lignin matrix. We also discuss dissociation mechanisms based on the calculated cross sections, available experimental data, virtual orbital analysis, and the knowledge on electron interactions with biomolecules. Our results point out a physical-chemical basis for electron-driven biomass delignification. The latter would be an essential step for efficient biofuel production from lignocellulosic materials.

Synthesis and Solar Cell Application of New Alternating Donor-Acceptor Copolymers Based on Variable Units of Fluorene, Thiophene, and Phenylene

A new series of donor acceptor copolymers were synthesized via the Witting route and applied as an active layer in organic thin-films solar cells. These copolymers are composed of fluorene thiophene and phenylene thiophene units. The ratio between those was systematically varied, and copolymers containing 0%, 50%, and 75% of phenylene thiophene were characterized and evaluated when used in photovoltaic devices. The copolymers' composition, photophysical, electrical, and morphological properties are addressed and correlated with device performance. The 50% copolymer ratio was found to be the best copolymer of the series, yielding a power conversion efficiency (PCE) under air mass (AM) 1.5 conditions of 2.4% in the bilayer heterojunction with the C-60 molecule. Aiming at flexible electronics applications, solutions based on the heterojunction of this copolymer with PCBM (6,6-phenyl-C-61-butyric acid methyl ester) were also successfully deposited using an inkjet printing method and used as an active layer in solar cells.

Photo- and electroluminescence in a series of PPV type terpolymers containing fluorene, thiophene and phenylene units

The emissive properties of terpolymers with fluorene, thiophene and phenylene groups, forming alternating PPV type structures, are discussed in terms of their composition, photo- and electroluminescence properties. The fluorene groups were inserted in each phenylene-vinylene and thiophene-vinylene units, and their concentration did not vary, representing 50% of the molar composition. The ratio of thiophene-vinylene/phenylene-vinylene varied in the range 25,50 and 75%. Photo- and electroluminescence properties were strongly dependent on the thiophene-vinylene content and were compared with the fluorene-vinylene-thiophene and fluorene-vinylene-phenylene parent copolymers. (C) 2012 Elsevier B.V. All rights reserved.

Photophysical and photovoltaic properties of a PPV type copolymer containing alternated fluorene and thiophene units

The synthesis and photophysical characterization of a PPV-type copolymer containing a fluorene derivative alternated with thiophene units is presented: poly(9,9'-dioctylfluorene-thiophene) (LAPPS29). Photophysical studies demonstrated that in the solid state only preformed ground state aggregates are responsible for exciton formation. These aggregates are formed with a wide range of size distribution. The emission from isolated segments is quenched either by resonant energy transfer, or by migration processes. Also, the main photovoltaic parameters are discussed in connection with the photophysical behavior.

Unraveling the structure of sugarcane bagasse after soaking in concentrated aqueous ammonia (SCAA) and ethanol production by Scheffersomyces (Pichia) stipitis

Abstract Background Fuel ethanol production from sustainable and largely abundant agro-residues such as sugarcane bagasse (SB) provides long term, geopolitical and strategic benefits. Pretreatment of SB is an inevitable process for improved saccharification of cell wall carbohydrates. Recently, ammonium hydroxide-based pretreatment technologies have gained significance as an effective and economical pretreatment strategy. We hypothesized that soaking in concentrated aqueous ammonia-mediated thermochemical pretreatment (SCAA) would overcome the native recalcitrance of SB by enhancing cellulase accessibility of the embedded holocellulosic microfibrils. Results In this study, we designed an experiment considering response surface methodology (Taguchi method, L8 orthogonal array) to optimize sugar recovery from ammonia pretreated sugarcane bagasse (SB) by using the method of soaking in concentrated aqueous ammonia (SCAA-SB). Three independent variables: ammonia concentration, temperature and time, were selected at two levels with center point. The ammonia pretreated bagasse (SCAA-SB) was enzymatically hydrolysed by commercial enzymes (Celluclast 1.5 L and Novozym 188) using 15 FPU/g dry biomass and 17.5 Units of β-glucosidase/g dry biomass at 50°C, 150 rpm for 96 h. A maximum of 28.43 g/l reducing sugars corresponding to 0.57 g sugars/g pretreated bagasse was obtained from the SCAA-SB derived using a 20% v/v ammonia solution, at 70°C for 24 h after enzymatic hydrolysis. Among the tested parameters, pretreatment time showed the maximum influence (p value, 0.053282) while ammonia concentration showed the least influence (p value, 0.612552) on sugar recovery. The changes in the ultra-structure and crystallinity of native SCAA-SB and enzymatically hydrolysed SB were observed by scanning electron microscopy (SEM), x-ray diffraction (XRD) and solid-state 13C nuclear magnetic resonance (NMR) spectroscopy. The enzymatic hydrolysates and solid SCAA-SB were subjected to ethanol fermentation under separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) by Scheffersomyces (Pichia) stipitis NRRL Y-7124 respectively. Higher ethanol production (10.31 g/l and yield, 0.387 g/g) was obtained through SSF than SHF (3.83 g/l and yield, 0.289 g/g). Conclusions SCAA treatment showed marked lignin removal from SB thus improving the accessibility of cellulases towards holocellulose substrate as evidenced by efficient sugar release. The ultrastructure of SB after SCAA and enzymatic hydrolysis of holocellulose provided insights of the degradation process at the molecular level.

Chemical composition and enzymatic digestibility of sugarcane clones selected for varied lignin content

Abstract Background The recalcitrance of lignocellulosic materials is a major limitation for their conversion into fermentable sugars. Lignin depletion in new cultivars or transgenic plants has been identified as a way to diminish this recalcitrance. In this study, we assessed the success of a sugarcane breeding program in selecting sugarcane plants with low lignin content, and report the chemical composition and agronomic characteristics of eleven experimental hybrids and two reference samples. The enzymatic digestion of untreated and chemically delignified samples was evaluated to advance the performance of the sugarcane residue (bagasse) in cellulosic-ethanol production processes. Results The ranges for the percentages of glucan, hemicellulose, lignin, and extractive (based on oven-dry biomass) of the experimental hybrids and reference samples were 38% to 43%, 25% to 32%, 17% to 24%, and 1.6% to 7.5%, respectively. The samples with the smallest amounts of lignin did not produce the largest amounts of total polysaccharides. Instead, a variable increase in the mass of a number of components, including extractives, seemed to compensate for the reduction in lignin content. Hydroxycinnamic acids accounted for a significant part of the aromatic compounds in the samples, with p-coumaric acid predominating, whereas ferulic acid was present only in low amounts. Hydroxycinnamic acids with ester linkage to the hemicelluloses varied from 2.3% to 3.6%. The percentage of total hydroxycinnamic acids (including the fraction linked to lignin through ether linkages) varied from 5.0% to 9.2%, and correlated to some extent with the lignin content. These clones released up to 31% of glucose after 72 hours of digestion with commercial cellulases, whereas chemically delignified samples led to cellulose conversion values of more than 80%. However, plants with lower lignin content required less delignification to reach higher efficiencies of cellulose conversion during the enzymatic treatment. Conclusion Some of the experimental sugarcane hybrids did have the combined characteristics of high biomass and high sucrose production with low lignin content. Conversion of glucan to glucose by commercial cellulases was increased in the samples with low lignin content. Chemical delignification further increased the cellulose conversion to values of more than 80%. Thus, plants with lower lignin content required less delignification to reach higher efficiencies of cellulose conversion during the enzymatic treatment.

Mapping the lignin distribution in pretreated sugarcane bagasse by confocal and fluorescence lifetime imaging microscopy

Abstract Background Delignification pretreatments of biomass and methods to assess their efficacy are crucial for biomass-to-biofuels research and technology. Here, we applied confocal and fluorescence lifetime imaging microscopy (FLIM) using one- and two-photon excitation to map the lignin distribution within bagasse fibers pretreated with acid and alkali. The evaluated spectra and decay times are correlated with previously calculated lignin fractions. We have also investigated the influence of the pretreatment on the lignin distribution in the cell wall by analyzing the changes in the fluorescence characteristics using two-photon excitation. Eucalyptus fibers were also analyzed for comparison. Results Fluorescence spectra and variations of the decay time correlate well with the delignification yield and the lignin distribution. The decay dependences are considered two-exponential, one with a rapid (τ1) and the other with a slow (τ2) decay time. The fastest decay is associated to concentrated lignin in the bagasse and has a low sensitivity to the treatment. The fluorescence decay time became longer with the increase of the alkali concentration used in the treatment, which corresponds to lignin emission in a less concentrated environment. In addition, the two-photon fluorescence spectrum is very sensitive to lignin content and accumulation in the cell wall, broadening with the acid pretreatment and narrowing with the alkali one. Heterogeneity of the pretreated cell wall was observed. Conclusions Our results reveal lignin domains with different concentration levels. The acid pretreatment caused a disorder in the arrangement of lignin and its accumulation in the external border of the cell wall. The alkali pretreatment efficiently removed lignin from the middle of the bagasse fibers, but was less effective in its removal from their surfaces. Our results evidenced a strong correlation between the decay times of the lignin fluorescence and its distribution within the cell wall. A new variety of lignin fluorescence states were accessed by two-photon excitation, which allowed an even broader, but complementary, optical characterization of lignocellulosic materials. These results suggest that the lignin arrangement in untreated bagasse fiber is based on a well-organized nanoenvironment that favors a very low level of interaction between the molecules.