Nonrecurrent MECP2 duplications mediated by genomic architecture-driven DNA breaks and break-induced replication repair

Recurrent submicroscopic genomic copy number changes are the result of nonallelic homologous recombination (NAHR). Nonrecurrent aberrations, however, can result from different nonexclusive recombination-repair mechanisms. We previously described small microduplications at Xq28 containing MECP2 in four male patients with a severe neurological phenotype. Here, we report on the fine-mapping and breakpoint analysis of 16 unique microduplications. The size of the overlapping copy number changes varies between 0.3 and 2.3 Mb, and FISH analysis on three patients demonstrated a tandem orientation. Although eight of the 32 breakpoint regions coincide with low-copy repeats, none of the duplications are the result of NAHR. Bioinformatics analysis of the breakpoint regions demonstrated a 2.5-fold higher frequency of Alu interspersed repeats as compared with control regions, as well as a very high GC content (53%). Unexpectedly, we obtained the junction in only one patient by long-range PCR, which revealed nonhomologous end joining as the mechanism. Breakpoint analysis in two other patients by inverse PCR and subsequent array comparative genomic hybridization analysis demonstrated the presence of a second duplicated region more telomeric at Xq28, of which one copy was inserted in between the duplicated MECP2 regions. These data suggest a two-step mechanism in which part of Xq28 is first inserted near the MECP2 locus, followed by breakage-induced replication with strand invasion of the normal sister chromatid. Our results indicate that the mechanism by which copy number changes occur in regions with a complex genomic architecture can yield complex rearrangements.

INFLUENCE OF ALTITUDE, AGE AND DIAMETER ON YIELD AND ALPHA-BISABOLOL CONTENT OF CANDEIA TREES (Eremanthus erythropappus)

The heartwood of candeia tree is a source of essential oil rich in alpha-bisabolol, a substance widely used in the cosmetic and pharmaceutical industry. Bearing in mind the economic importance of alpha-bisabolol, this work aimed to evaluate the influence of tree age on the yield and content of alpha-bisabolol present in essential oil from candeia, considering two distinct reliefs and three diameter classes, in Aiuruoca region, south Minas Gerais state. The two distinct reliefs correspond respectively to one section of the stand growing at 1,000m of altitude (Area 1) and another section growing at 1,100m of altitude (Area 2). In each section, 15 trees were felled from among 3 different diameter classes. Discs were removed from the base of each tree to estimate their age by doing growth ring count. Soil samples were taken and Subjected to physical and chemical analysis. The logs were reduced into chips and random samples were taken for distillation to extract essential oil. The method used was steam distillation at a pressure of 2 kgf/cm(2)/2.5 h. The chemical analysis was performed in a gas chromatograph (GC) based on the alpha-bisabolol standard reference. The yield of essential oil from trees in Area I was higher than that from trees in Area 2, with the same pattern of influence for older trees. In Area 2, the alpha-bisabolol content was higher in younger trees. No differences were found between the relevant parameters in relation to diameter classes.

Composition of phenolic acids content in apple (Malus sp) pomace

The present work aimed the study of phenolic acids composition in apple pomace of Gala and Fuji cultivars. Phenolic acids were fractionated in phenolic acids, esterified and insoluble and analyzed by gas chromatography-mass spectrometry (GC-MS). Sixteen phenolic acids were identified in apple pomace samples. Total phenolic acids in apple pomace from Gala and Fuji cultivars were, in dry weight, 93.94 mg/g and 68.38 mg/g, respectively. Content of free phenolic acids in apple pomace from Gala cultivar was 29.11 mg/g and the following acids were identified: salicylic, protocatequinic, quinic, p-coumaric, gallic, propylgallate and synapic. Content of free phenolic acids in apple pomace from Fuji cultivar was 16.03 mg/g and the following acids were identified: salicylic, protocatequinic, gallic, ferulic and sinapic. Salicylic was the predominant free phenolic acids found in both cultivars, consisting of 91.67% and 63.57% of the free phenolic acids in Gala and Fuji cultivars, respectively. Chlorogenic acid (1.147 mg/g) was found only in apple pomace from Fuji cultivar. Content of esterified phenolic acids in apple pomace from Gala and Fuji cultivars were 53.75 mg/g and 48.29 mg/g, respectively. It was verified that the predominant esterified phenolic acid in pomace from apple Gala is derived from salicylic acid (52.76 mg/g). Acids derived from gallic acid (0.175 mg/g), propylgallate acid (0.198 mg/g), ferulic acid (0.159 mg/g) and sinapic acid (0.140 mg/g) were also found in Gala cultivar. Regarding to pomace from cultivar Fuji, the main esterified phenolic acid found is also derived from salicylic acid (47.42 mg/g) followed by gallic acid (0.270 mg/g), benzoic acid (0.194 mg/g) and sinapic acid (0.115 mg/g). Content of insoluble phenolic acids in apple pomace from Gala and Fugi cultivars were, in dry weight, 11.08 mg/g and 4.05 mg/g, respectively Insoluble phenolic acids derived from salicylic acid were found in higher concentrations in apple pomace from both cultivars.

An approach to chemotaxonomy to the fatty acid content of some Malvaceae species

Malvales is an order of flowering plants with a controversial circumscription. The relationships between taxa, particularly Malvaceae, Bombacaceae, Sterculiaceae, and Tiliaceae, are not well delineated. Several studies have reported the fatty acid compositions of Malvaceae plants but not for taxonomic purposes. In the present study, the fatty acid composition of oilseeds from seven species belonging to the Malvaceae family was determined by capillary gas chromatography/mass spectrometry (GC/MS), and the quantitative distribution of fatty acids was analyzed by a cluster analysis With Euclidean Distance and UPGMA. The oil content in the seeds was very low (8.3-11.8%). The profile of fatty acids showed that there were two distinct groups: species rich in palmitic acid (Herissantia tiubae, Sidastrum paniculatum and Sida rhombifolia) and species rich in linoleic acid (other Sida species). The fatty acid profiles found for Sida species are consistent with other reported data. Although our data support a distinction between Sida and Sidastrum, more species should be analyzed to evaluate the real taxonomic value of differences in fatty acid content for distinguishing Malvaceae. (C) 2010 Elsevier Ltd. All rights reserved.

Quantitative ester analysis in cachaca and distilled spirits by gas chromatography-mass spectrometry (GC-MS)

An analytical procedure for the separation and quantification of ethyl acetate, ethyl butyrate, ethyl hexanoate, ethyl lactate, ethyl octanoate, ethyl nonanoate, ethyl decanoate, isoamyl octanoate, and ethyl laurate in cachaca, rum, and whisky by direct injection gas chromatography-mass spectrometry was developed. The analytical method is simple, selective, and appropriated for the determination of esters in distilled spirits. The limit of detection ranged from 29 (ethyl hexanoate) to 530 (ethyl acetate) mu g L-1, whereas the standard deviation for repeatability was between 0.774% (ethyl hexanoate) and 5.05% (isoamyl octanoate). Relative standard deviation values for accuracy vary from 90.3 to 98.5% for ethyl butyrate and ethyl acetate, respectively. Ethyl acetate was shown to be the major ester in cachaca (median content of 22.6 mg 100 mL(-1) anhydrous alcohol), followed by ethyl lactate (median content of 8.32 mg 100 mL(-1) anhydrous alcohol). Cachaca produced in copper and hybrid alembic present a higher content of ethyl acetate and ethyl lactate than those produced in a stainless-steel column, whereas cachaca produced by distillation in a stainless-steel column present a higher content of ethyl octanoate, ethyl decanoate, and ethyl laurate. As expected, ethyl acetate is the major ester in whiskey and rum, followed by ethyl lactate for samples of rum. Nevertheless, whiskey samples exhibit ethyl lactate at contents lower or at the same order of magnitude of the fatty esters.

A method to determine volatile contaminants in polyethylene terephthalate (PET) packages by HDC-GC-FID and its application to post-consumer materials

A simple and low cost method to determine volatile contaminants in post-consumer recycled PET flakes was developed and validated by Headspace Dynamic Concentration and Gas Chromatography-Flame Ionization Detection (HDC-GC-FID). The analytical parameters evaluated by using surrogates include: correlation coefficient, detection limit, quantification limit, accuracy, intra-assay precision, and inter-assay precision. In order to compare the efficiency of the proposed method to recognized automated techniques, post-consumer PET packaging samples collected in Brazil were used. GC-MS was used to confirm the identity of the substances identified in the PET packaging. Some of the identified contaminants were estimated in the post-consumer material at concentrations higher than 220 ng.g-1. The findings in this work corroborate data available in the scientific literature pointing out the suitability of the proposed analytical method.

Deleterious effect of TRIS buffer on growth rates and pigment content of Gracilaria birdiae Plastino & E.C. Oliveira (Gracilariales, Rhodophyta)

This work evaluated the effects of Tris (hydroxymethyl)-aminomethane (TRIS) buffer and its interaction with nutrient concentration on the development of Gracilaria birdie, a common species on the Brazilian coast that has been exploited for agar production. Responses to different conditions were assessed through growth rates and pigment content (chlorophyll a, phycoerythrin, phycocyanin and allophycocyanin). Provasoli's nutrient solution with and without TRIS addition was tested at concentrations of 12.5, 25 and 50%. The pH was also monitored. G. birdiae grew better in the absence of TRIS and at low nutrient concentrations, 12.5 and 25% (growth rates of 10.8-11.3%.day-1). Higher contents of phycoerythrin and chlorophyll a were observed without TRIS at 12.5 and 25% (Phycoerythrin, 649.6-698.0 μg g-1 fresh biomass; Chlorophyll a, 156.0-168.6 μg g-1 fresh biomass). These findings highlight the deleterious effect of TRIS on growth and phycoerythrin and chlorophyll a content. They also demonstrate the importance of appropriate nutrient concentration for laboratory cultures, depending on the intrinsic characteristics of each species.

Comparação de técnicas de determinação de ésteres em cachaça

An analytical comparison of three different techniques for quantitative profile of esters in cachaça is reported. The Ministério da Agricultura Pecuária e Abastecimento (MAPA) recommends the use of GC/FID or volumetry. Despite being laborious and lacking in chemical speciation, the volumetric technique for total ester content shows to be appropriate, reproducible, and accurate for the analysis of cachaça. However, the GC/FID suggested by MAPA, considering only ethyl acetate, shows inaccuracy, underestimating the total ester content by a median factor of 72%, mainly due to the absence of ethyl lactate analysis. On the other hand, the GC/MS technique that comprises the analysis and speciation of nine esters, including ethyl lactate, proved to be reproducible, simple, fast and accurate for the analysis of total ester content in cachaça. Thus, the total ester content results obtained using GC/FID must be considered with precaution.

Teores de carbamato de etila em aguardentes de cana e mandioca: parte II

Herein, we report the concentration of ethyl carbamate (EC) and copper in 380 samples of sugar-cane spirit and 45 samples of manioc spirit as determined by GC-MS and FAAS respectively. The cyanide content determined spectrophotometrically is reported for the manioc spirit. Sugar cane spirit produced by alembic distillation (70,0 µg L-1) shown a lower content of EC than samples produced by column distillation (270 µg L-1). No simple correlation between the content of EC and copper for sugar cane spirit as well among the concentration of EC, copper, and cyanide for manioc spirit could be observed.

Multivariate analysis of the effects of soil parameters and environmental factors on the flavonoid content of leaves of Passiflora incarnata L., Passifloraceae

The aim of the present study was to evaluate the effect of soil characteristics (pH, macro- and micro-nutrients), environmental factors (temperature, humidity, period of the year and time of day of collection) and meteorological conditions (rain, sun, cloud and cloud/rain) on the flavonoid content of leaves of Passiflora incarnata L., Passifloraceae. The total flavonoid contents of leaf samples harvested from plants cultivated or collected under different conditions were quantified by high-performance liquid chromatography with ultraviolet detection (HPLC-UV/PAD). Chemometric treatment of the data by principal component (PCA) and hierarchic cluster analyses (HCA) showed that the samples did not present a specific classification in relation to the environmental and soil variables studied, and that the environmental variables were not significant in describing the data set. However, the levels of the elements Fe, B and Cu present in the soil showed an inverse correlation with the total flavonoid contents of the leaves of P. incarnata.

Mineral and vitamin content of beef, chicken, and turkey hydrolysates mineral and vitamin content of protein hydrolysates

The purpose of this study was to assess the concentration of vitamins and minerals in meat protein hydrolysates. Calcium, phosphorus and iron were analyzed by inductively coupled-plasma atomic emission spectrophotometry; vitamin C was analyzed by the reduction of cupric ions and vitamins B1 and B2 by fluorescence. Regarding minerals, the beef hydrolysate (BH) had more iron than the turkey hydrolysate (TH) and the chicken hydrolysate (CH); TH had a little more phosphorus. BH had the largest amount of vitamin C, and similar amounts of vitamins B1 and B2. The amount of these nutrients found in the hydrolysates suggests that it is possible to use them to enrich special dietary formulations.

Energy and amino acid content in phase 1 nursery diet: piglet performance and body chemical composition

It was evaluated the effects of metabolizable energy (ME) and digestible lysine (dLYS) densities on performance and body composition of weaned piglets. The study used 114 piglets weaned at 7.4 ± 0.80 kg, out of which 108 were allotted in the nursery and 6 were slaughtered on the weaning day to determine comparative data of body chemical composition. Six nutrients densities were stipulated from a previous study based on the highest nitrogen retention, maintaining the following ME:LYS relationship in the experimental diets: 3,390:1.291; 3,450:1.409; 3,650:1.411; 3,780:1.461; 3,940:1.507; and 4,109 kcal/kg ME:1.564% dLYS. The experimental diets were offered for 13 days when the piglets reached 12.986 ± 1.449 kg of body weight. The probable residual effects of nutritional density on the subsequent performance of the piglets were evaluated. At the end of initial phase 1, six piglets from each density were slaughtered to determine their chemical composition in body fractions and empty body. There was no significant influence of nutritional levels on the performance of the piglets at the end of the evaluation. The results of food conversion and body composition confirm the level indicated in the previous study, 4 g dLYS/Mcal of ME. The increase of energy and lysine densities confirms the need for a correct relationship among both of them to assure better performance of the piglets at the beginning of the growing phase.

Sugar and total energy content of household food purchases in Brazil

Objective To study the role of energy derived from sugar (both table sugar and sugar added to processed foods) in the total energy content of food purchases in Brazil.Design Food purchase data were collected during a national household budget survey carried out between June 2002 and July 2003 on a probabilistic sample representative of all households in the country. The amount of food purchased in this 12-month period was transformed into energy and energy from sugar using food composition tables. Multiple linear regression models were used to study the association between amount of energy from sugar and total energy content of food purchases, controlling for sociodemographic variables and potential interactions between these variables and sugar purchases.Results There was a positive and significant association between energy from sugar and total household energy purchases. A 1 kJ increase in sugar purchase corresponded to a 3·637 kJ increase in total energy. In the absence of expenditure on meals outside the home, i.e. when household food purchases tend to approximate actual food consumption by household members, sugar purchase of 1926·35 kJ/d (the 90th percentile of the distribution of sugar purchases in Brazil) was associated, depending on income strata, with total energy purchase over 40\201360 per cent of the recommended daily value for energy intake in Brazil.Conclusions The present results corroborate the recommendations of the WHO and the Brazilian Ministry of Health regarding limiting the consumption of sugar

Recovery and upgrading bovine rumen protein by extrusion: effect of lipid content on protein disulphide cross-linking, solubility and molecular weight

Bovine rumen protein with two levels of residual lipids (1.9 per cent or 3.8 per cent) was subjected to thermoplastic extrusion under different temperatures and moisture contents. Protein solubility in different buffers, disulphide cross-linking and molecular weight distribution were determined on the extrudates. After extrusion, samples with 1.9 per cent residual lipids content had a higher concentration of protein insoluble by undetermined forces, irrespective of feed moisture and processing temperature used. Lipid content of 3.8 per cent in the feed material resulted in more protein participating in the extrudate network through non-covalent interactions (hydrophobic and electrostatic) and disulphide bonds. A small dependency of the extrusion process on moisture and temperature and a marked dependency on lipid content, especially phospholipid, was observed, Electrophoresis under non-reducing conditions showed that protein extrusion with low feed moisture promoted high molecular breakdown inside the barrel, probably due to intense shear force, and further protein aggregation at the die end

NEAR INFRARED SPECTROSCOPY FOR ESTIMATING SUGARCANE BAGASSE CONTENT IN MEDIUM DENSITY FIBERBOARD

Medium density fiberboard (MDF) is an engineered wood product formed by breaking down selected lignin-cellulosic material residuals into fibers, combining it with wax and a resin binder, and then forming panels by applying high temperature and pressure. Because the raw material in the industrial process is ever-changing, the panel industry requires methods for monitoring the composition of their products. The aim of this study was to estimate the ratio of sugarcane (SC) bagasse to Eucalyptus wood in MDF panels using near infrared (NIR) spectroscopy. Principal component analysis (PCA) and partial least square (PLS) regressions were performed. MDF panels having different bagasse contents were easily distinguished from each other by the PCA of their NIR spectra with clearly different patterns of response. The PLS-R models for SC content of these MDF samples presented a strong coefficient of determination (0.96) between the NIR-predicted and Lab-determined values and a low standard error of prediction (similar to 1.5%) in the cross-validations. A key role of resins (adhesives), cellulose, and lignin for such PLS-R calibrations was shown. PLS-DA model correctly classified ninety-four percent of MDF samples by cross-validations and ninety-eight percent of the panels by independent test set. These NIR-based models can be useful to quickly estimate sugarcane bagasse vs. Eucalyptus wood content ratio in unknown MDF samples and to verify the quality of these engineered wood products in an online process.