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Background: Parkinson's disease (PD) is the second most common neurodegenerative disorder, affecting millions of people. Genome-wide association studies (GWAS) have found >25 genetic risk factors and at least 15 loci directly associated with PD. Recent advances in Next Generation DNA Sequencing technologies, such as the semiconductor-based Ion Torrent platform, make multigene sequencing cheaper, faster, and more reliable. Objectives: Our objective was to test the power of the Next Generation Sequencing technology to analyze a large cohort samples of PD patients from Southern Italy by screening the majority of the most relevant PD-related genes known for single and compound mutations. Methods: To achive a rapid,robust, and cost-effective genetic analysis of a PD cohort, we designed a multiplex, amplicon based gene panel made by 42 genes. We conducted parallel sequencing using the Ion Torrent Personal Genome Machine (PGM)(®) system to detect mutations in 42 blood DNA samples from PD patients. To process data from PGM Ion Torrent runs, Ion Torrent Suite (TS) software v. 5.10 was used. Annotation was made using ANNOVAR and the variants where priorized using a standard filtering pipeline. Results: After bioinformatics analysis and filtering, 98% coverage of the targeted regions was obtained with at least >200-fold mean depth. We detected 50 coding nonsynonymous variants (indels, single-nucleotide variations (SNVs) and frameshift variants with a MAF

Introduction:Inherited Peripheral Neuropathies.represent a large group of hereditary diseases that comprises the hereditary motor and sensory neuropathy, more known as Charcot-Marie-Tooth (CMT), hereditary sensory and autonomic neuropathy (HSNA) and distal motor neuropathy (dHMN). Typically IPNs are characterized by distal wasting and weakness, decreased deep tendon reflexes, contractures and skeletal deformities. More than 60 nuclear genes are implicated in the pathogenesis.In the last few years, the molecular diagnosis improved with the advent of NGS, which is now the strategy used in routine in many laboratories. Aim of the study We performed genetic tests for 49 genetically un-diagnosed IPNs patients who had been excluded 17p12 (PMP22) duplication by Real Time PCR and microsatellite markers. Materials and Methods: We designed a genes panel using the Ion Ampliseq Designer (Thermo Fisher Scientific) targeting the coding sequence comprising the intron/exon boundaries of 32 genes associated with IPNs phenotype, reaching a target coverage of 99,13%. The amplicon libraries obtained by the Ion AmpliSeq Library Kit 2.0 (Thermo Fisher Scientific), were subsequently barcoded, pool together in equimolar concentrations and enriched using the One Touch 2 instrument. Sequencing was performed on an Ion PGM machine using an Ion 318 sequencing chip. The obtained raw data were analyzed with the Ion Torrent Suite 5.10 for the alignment and variants caller. To annotate the variants ANNOVAR tool was also used. The identified variants were confirmed by Sanger sequencing. Results: By using IPNs genes panel, we are able to detect in 19 out of 49 examined patients pathogenic variants in the following genes: KIF1B, MFN2, MPZ, SETX, SBF2, DCTN1, GARS, GAN, DNM2, WINK1 and FIG4. KIF1B and MFN2 genes result to be, in our study, the most frequent mutated genes. Conclusions: Targeted gene panel sequencing is a power tool to resolve uncertain cases with suspected IPN diagnosis. It strikingly reduces the cost and the time with better detection efficiency; but, because the number of causative genes for IPNs is still growing, it is necessary to have a continuous updating of the involved genes to facilitate the diagnosis of the unsolved cases.

Introduction: Vanishing white matter disease (VWM) is an autosomal recessive leukodystrophy caused by mutations genes encoding the translational initiation factor 2B: EIF2B1-EIFB5. It is a heterogeneous neurological disorder with prominent cerebellar ataxia and spasticity in childhood-onset form and behavioural changes, dementia and seizures in adult-onset form (#15% of cases). The course of the disease is chronic progressive with episodes of stress-provoked rapid neurological deterioration. Purpose of the study: We present the case of a 60 years old patient, MTHFR homozygous, presenting white matter lesions and a subcortical vascular encephalopathy being diagnosed. To investigate a possible genetic cause, suspecting CADASIL, genetic and genomic analysis were performed. Materials and Methods: For genetic and genomic investigations, Sanger sequencing of NOTCH3 gene, linked to CADASIL disease, and Next Generation Sequencing (NGS) analysis were performed on the patient. NOTCH3 sequencing was performed using primer pairs for all 22 exons, while for NGS analysis, we used a genes panel including 753 genes involved in neurological diseases. Libraries were prepared using Ion Ampliseq Library preparation kit 2.0 (Thermo Fisher Scientific) and they were run on Ion Torrent PGM (Thermo Fisher Scientific). We performed the alignment using Torrent Suite v. 5.10 and we used ANNOVAR for annotation step. Results: Sanger sequencing did not reveal any mutation in NOTCH3 gene, excluding CADASIL disease as the genetic cause responsible for the clinical history of our patient. NGS detected a novel heterozygous non- synonimous variant in exon 5 of EIF2B3 gene (c. C554G; p. S185C), whose mutations have been strongly related to leukoencephalopathy and VWM disease. Furthermore, all in silico analysis conducted on this variant predicts its deleterious impact that could affect protein function. Conclusion: S185C variant in EIF2B3 gene was discovered in an atypical condition of adult onset Leukodistrophy, a disease that usually occurs in childhood. It is uncertain if S185C could be responsible for the clinical history of our patient or if the peculiarity of his symptoms could be related to the potential contribution of this new detected variant on the phenotypical spectrum observed in the disease.

Made available in DSpace on 2022-04-28T19:27:27Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-07-08 Background and Aims: Juvenile-to-adult phase transition is marked by changes in leaf morphology, mostly due to the temporal development of the shoot apical meristem, a phenomenon known as heteroblasty. Sugars and microRNA-controlled modules are components of the heteroblastic process in Arabidopsis thaliana leaves. However, our understanding about their roles during phase-changing in other species, such as Passiflora edulis, remains limited. Unlike Arabidopsis, P. edulis (a semi-woody perennial climbing vine) undergoes remarkable changes in leaf morphology throughout juvenile-to-adult transition. Nonetheless, the underlying molecular mechanisms are unknown. Methods: Here we evaluated the molecular mechanisms underlying the heteroblastic process by analysing the temporal expression of microRNAs and targets in leaves as well as the leaf metabolome during P. edulis development. Key Results: Metabolic profiling revealed a unique composition of metabolites associated with leaf heteroblasty. Increasing levels of glucose and α-trehalose were observed during juvenile-to-adult phase transition. Accumulation of microRNA156 (miR156) correlated with juvenile leaf traits, whilst miR172 transcript accumulation was associated with leaf adult traits. Importantly, glucose may mediate adult leaf characteristics during de novo shoot organogenesis by modulating miR156-targeted PeSPL9 expression levels at early stages of shoot development. Conclusions: Altogether, our results suggest that specific sugars may act as co-regulators, along with two microRNAs, leading to leaf morphological modifications throughout juvenile-to-adult phase transition in P. edulis. Departamento de Biologia Vegetal/Instituto de Biotecnologia Aplicada A Agropecuária (BIOAGRO) Universidade Federal de Viçosa Universidade Estadual Do Maranhão Departamento de Biologia Vegetal Universidade Federal de Viçosa Instituto de Educação Agricultura e Ambiente Universidade Federal Do Amazonas Escola Superior de Agricultura Luiz de Queiroz Universidade de São Paulo Instituto de Biociências Universidade Estadual de São Paulo Instituto de Biociências Universidade Estadual de São Paulo

This study aimed to prepare hydrogel containing Cymbopogon citratus (DC.) Stapf, Poaceae, volatile oil encapsulated in poly (d,l-lactide-co-glycolide) nanoparticles and to evaluate its in vitro anti-herpetic activity. Polymeric nanoparticles were prepared by solvent emulsification-diffusion method and incorporated in carbomer hydrogels. In vitro release profiles for the nanogel, loaded nanoparticles and hydrogel containing free oil were evaluated by dialysis. Inhibitory activities against Herpes simplex for the formulations were investigated in Vero cells. Hydrogel was developed using nanoparticles with mean diameter of 217.1 nm and negative Zeta potential (−20.5 mV). Volatile oil release profile showed a biphasic pattern with an initial faster release and subsequent sustained phase in all formulations. Nanogel strongly inhibited virus in a non-cytotoxic concentration, 42.16 times lower than free oil, 8.76 and 2.23 times than loaded nanoparticles and hydrogel containing free oil, respectively. These results highlight the potential of nanogel to protect oil against volatilization, control release and improve its anti-herpetic activity. Keywords: Nanotechnology, Polymeric nanoparticles, Volatile oil, Lemongrass, Hydrogel, Herpes simplex virus

Impaired lysine biosynthesis in dapat mutant simulates a stress response culminating in metabolic reprogramming, such that alternative substrates support energy generation once carbohydrate metabolism is down-regulated., Lysine (Lys) connects the mitochondrial electron transport chain to amino acid catabolism and the tricarboxylic acid cycle. However, our understanding of how a deficiency in Lys biosynthesis impacts plant metabolism and growth remains limited. Here, we used a previously characterized Arabidopsis mutant (dapat) with reduced activity of the Lys biosynthesis enzyme L,L-diaminopimelate aminotransferase to investigate the physiological and metabolic impacts of impaired Lys biosynthesis. Despite displaying similar stomatal conductance and internal CO2 concentration, we observed reduced photosynthesis and growth in the dapat mutant. Surprisingly, whilst we did not find differences in dark respiration between genotypes, a lower storage and consumption of starch and sugars was observed in dapat plants. We found higher protein turnover but no differences in total amino acids during a diurnal cycle in dapat plants. Transcriptional and two-dimensional (isoelectric focalization/SDS-PAGE) proteome analyses revealed alterations in the abundance of several transcripts and proteins associated with photosynthesis and photorespiration coupled with a high glycine/serine ratio and increased levels of stress-responsive amino acids. Taken together, our findings demonstrate that biochemical alterations rather than stomatal limitations are responsible for the decreased photosynthesis and growth of the dapat mutant, which we hypothesize mimics stress conditions associated with impairments in the Lys biosynthesis pathway.

Adenine nucleotides are essential in countless processes within the cellular metabolism. In plants, ATP is mainly produced in chloroplasts and mitochondria through photophosphorylation and oxidative phosphorylation, respectively. Thus, efficient adenylate transport systems are required for intracellular energy partitioning between the cell organelles. Adenylate carriers present in different subcellular compartments have been previously identified and biochemically characterized in plants. Here, by using data-mining bioinformatics tools, we propose how, and to what extent, these carriers integrate energy metabolism within a plant cell under different environmental conditions. We demonstrate that the expression pattern of the corresponding genes is variable under different environmental conditions, suggesting that specific adenylate carriers have distinct and nonredundant functions in plants.

The main characteristic of sponges (Porifera) is the presence of the aquiferous system-a system formed by canals and choanocyte chambers, in which the sponges carry out most of their physiological functions. Despite of the importance for the biology of the group, the knowledge about this structure is still incipient, even when morphological investigations are taken in account. Here, we investigated the anatomy and ultrastructure of the tropical demosponge Cladocroce caelum (Haplosclerida, Demospongiae) using light and electron microscopy. In the studied region, specimens of this species were repent or repent-branched, possessing one to several oscula. A uniform and reduced atrium was found just below each osculum. There was a thin ectosome and the choanosome presented meager mesohyl, but a high number of choanocyte chambers. The choanocyte chambers were rounded, and, as in other haplosclerids, they are found separated from the mesohyl by endopinacocytes, "hanging" in the inhalant canals. Even though the utility of the general organization of the aquiferous system has been advocated as a possible tool to understand the phylogeny of the group, we found that these characters might not be as useful as expected. The size of the particles ingested by the sponge and the amount of bacteria to sustain their bodies are discussed. In addition, we found that the density of choanocyte chambers was reduced when the specimens were carrying out the spermatogenesis, indicating that the reproduction may impair the filtering activity of the sponge. Our findings consist in a first step to better comprehend the physiology, development, and adaptation to the environmental conditions where the species is found.

Plant respiration mostly depends on the activity of glycolysis and the oxidation of organic acids in the tricarboxylic acid cycle to synthesize ATP. However, during stress situations plant cells also use amino acids as alternative substrates to donate electrons through the electron-transfer flavoprotein (ETF)/ETF:ubiquinone oxidoreductase (ETF/ETFQO) complex to the mitochondrial electron transport chain (mETC). Given this, we investigated changes of the oxidative phosphorylation (OXPHOS) system in Arabidopsis thaliana cell culture under carbohydrate starvation supplied with a range of amino acids. Induction of isovaleryl-CoA dehydrogenase (IVDH) activity was observed under carbohydrate starvation which was associated with increased amounts of IVDH protein detected by immunoblotting. Furthermore, activities of the protein complexes of the mETC were reduced under carbohydrate starvation. We also observed that OXPHOS system activity behavior is differently affected by different amino acids and that proteins associated with amino acids catabolism are upregulated in cells following carbohydrate starvation. Collectively, our results support the contention that ETF/ETFQO is an essential pathway to donate electrons to the mETC and that amino acids are alternative substrates to maintain respiration under carbohydrate starvation.

The tricarboxylic acid (TCA) cycle, a crucial component of respiratory metabolism, is composed of a set of eight enzymes present in the mitochondrial matrix. However, most of the TCA cycle enzymes are encoded in the nucleus in higher eukaryotes. In addition, evidence has accumulated demonstrating that nuclear genes were acquired from the mitochondrial genome during the course of evolution. For this reason, we here analyzed the evolutionary history of all TCA cycle enzymes in attempt to better understand the origin of these nuclear-encoded proteins. Our results indicate that prior to endosymbiotic events the TCA cycle seemed to operate only as isolated steps in both the host (eubacterial cell) and mitochondria (alphaproteobacteria). The origin of isoforms present in different cell compartments might be associated either with gene-transfer events which did not result in proper targeting of the protein to mitochondrion or with duplication events. Further in silico analyses allow us to suggest new insights into the possible roles of TCA cycle enzymes in different tissues. Finally, we performed coexpression analysis using mitochondrial TCA cycle genes revealing close connections among these genes most likely related to the higher efficiency of oxidative phosphorylation in this specialized organelle. Moreover, these analyses allowed us to identify further candidate genes which might be used for metabolic engineering purposes given the importance of the TCA cycle during development and/or stress situations.

The adequacy of Stribeck curves for the characterization of the lubricity of biodiesels, B20 blends and diesel fuel is demonstrated. Ball-on-disk tests in the speed range 2-1570 mm/s were performed to obtain the Stribeck plots and the results are compared to those from the conventional HFRR (high frequency reciprocating rig) ball-on-disk method (ASTM (American Society for Testing and Materials) D6079). Contrarily to the HFRR method, in the Stribeck tests very clear ball wear marks are seen without significant wear of the flat counterbody. These characteristics provide more confidence in the lubricity assessment of fuels. Moreover, in the Stribeck method higher lubricity at 60 degrees C is revealed for all biodiesels and the respective blends, related to the formation of protective oxide tribolayers. When evaluated by the HFRR test method, such temperature effect on the lubricity performance is not identified. From the point of view of the energy loss of the system, or friction response, lubricity is also better depicted by the Stribeck test method. The friction coefficient plots reveal that the major difference among the fuels occurs in the low velocity range, or in the start stop stage of moving components, where the poorest lubricity is attained with the neat diesel fuel and the best lubricity with the animal fat biodiesel. (C) 2014 Elsevier Ltd. All rights reserved.

The anti-HMPV (human metapneumovirus) activity was determined for sulfated dl -hybrid galactans obtained from the red seaweed Cryptonemia seminervis and their depolymerized products obtained by reductive partial hydrolysis. Structural studies carried out in three homogeneous depolymerized fractions DS-1, DS-2e and DS-3 (Mw of 51.6–63.8 kDa) showed that these galactans present different chemical characteristics, as monosaccharide composition, content of sulfate groups (14.1–29.9%) and agaran:carrageenan molar ratio diads, 2.7:1 for DS-1 and DS-2e and 1:1 for DS-3. The sulfate groups are located principally on C-2 of β- d -galactopyranose and 4,6-O-(1′-carboxyethylidene)-β- d -galactopyranose residues and on C-6 of α-galactose residues. Sulfated dl -galactans and their depolymerized products exhibited antiviral activity at a very early stage of the viral infection cycle. All fractions, except DS-2e inhibited HMPV replication by binding to the viral particle. Besides depolymerized galactans DS-2e and DS-3 inhibited the recognition of cell receptor by HMPV and penetration to the host cell, respectively.

Although microsatellites are the most popular molecular markers for population genetic studies, only a few loci have been characterized for sponges (Porifera). In the present work, we describe the isolation and characterization of seven polymorphic microsatellite loci from an enriched library of the marine calcareous sponge Clathrina aurea. This species is considered endemic to Brazil, where it is widely distributed. However, it was recently found in Peruvian waters. Seven loci were tested in 30 individuals of one population in southeastern Brazil (Cabo Frio). The number of alleles ranged from 5 to 18 (average 12.3 ± 3.9). The species presented high average values of observed and expected heterozygosities: 0.672 ± 0.25 and 0.837 ± 0.08, respectively. No linkage disequilibrium between all pairs of loci was found, but two loci in this population showed a significant excess of homozygotes. This is the second set of loci ever characterized for a calcareous sponge. These loci will be useful for population genetics studies among Brazilian populations, to elucidate the possible introduction of this species in the Pacific Ocean and to analyze other still poorly investigated issues in sponges.

The calcarean sponge Paraleucilla magna is classified as being an invasive species on the Mediterranean Sea, where it causes economic damages to mollusc farms. On the Brazilian coast, this species is considered to be cryptogenic, and information on its ecology is scarce. The same is true for Sycettusa hastifera, another calcarean sponge with a worldwide distribution. Data on the ecology of these species could help in elucidating their potential to become a threat if they are found to be exotic species in Brazil. In the present work, we studied habitat selection, growth and mortality of early juveniles of P. magna and habitat selection of S. hastifera in a Marine Reserve from Southeastern Brazil, where these species are abundant in the benthic community. Granite plates were used for habitat selection analysis, varying in substrate inclination (vertical and horizontal) and exposure to light and hydrodynamism (exposed and sheltered). To analyse the growth and mortality rates, sponges were mapped and then measured once a week for 10 weeks. If a monitored sponge was not found in the following week, it was considered to be dead. Our results showed that, although P. magna and S. hastifera are capable of inhabiting substrates exposed to different environmental conditions, they showed habitat preferences. Growth of the juveniles of P. magna seemed not to have damaged any neighbouring invertebrates. The mortality of juveniles of this species was higher during the first 2 weeks of life but its causes could not be elucidated.

Under heterotrophic conditions, carbohydrate oxidation inside the mitochondrion is the primary energy source for cellular metabolism. However, during energy-limited conditions, alternative substrates are required to support respiration. Amino acid oxidation in plant cells plays a key role in this by generating electrons that can be transferred to the mitochondrial electron transport chain via the electron transfer flavoprotein/ubiquinone oxidoreductase system. Autophagy, a catabolic mechanism for macromolecule and protein recycling, allows the maintenance of amino acid pools and nutrient remobilization. Although the association between autophagy and alternative respiratory substrates has been suggested, the extent to which autophagy and primary metabolism interact to support plant respiration remains unclear. To investigate the metabolic importance of autophagy during development and under extended darkness, Arabidopsis (

Secondary metabolites isolated from Simira eleiezeriana and Simira glaziovii were evaluated against herpes simplex virus (HSV-1) and (HSV-2). The 50% effective concentrations values (EC50) were calculated from the dose-response curve and the selectivity index (SI) against the virus. The physicochemical data LogP, (PSA), (NRB), (HBA) and (HBD) were obtained using Marvin Sketch. Among the tested compounds, conipheraldeyde, harman and simirane A showed better results with EC50 6.39; 4.90; 4.61 µg/mL and SI 78.3; 11.8; 7.01, respectively, for HSV-1, and EC50 41.2; 71.8; 3.73 µg/mL and SI 12.1; 24.7; 8.7, respectively, for HSV-2. The percentage of inhibition (PI) obtained for HSV-1 were higher than 60%, and for HSV-2 these compounds showed PI > 90%. The physical chemical data showed that the most active compounds satisfy the attributes for drugs with good oral bioavailability.

Cyanobacteria is a remarkable group of prokaryotic photosynthetic microorganisms, with several genera capable of fixing atmospheric nitrogen (N2) and presenting a wide range of morphologies. Although the nitrogenase complex is not present in all cyanobacterial taxa, it is spread across several cyanobacterial strains. The nitrogenase complex has also a high theoretical potential for biofuel production, since H2 is a by-product produced during N2 fixation. In this review we discuss the significance of a relatively wide variety of cell morphologies and metabolic strategies that allow spatial and temporal separation of N2 fixation from photosynthesis in cyanobacteria. Phylogenetic reconstructions based on 16S rRNA and nifD gene sequences shed light on the evolutionary history of the two genes. Our results demonstrated that (i) sequences of genes involved in nitrogen fixation (nifD) from several morphologically distinct strains of cyanobacteria are grouped in similarity with their morphology classification and phylogeny, and (ii) nifD genes from heterocytous strains share a common ancestor. By using this data we also discuss the evolutionary importance of processes such as horizontal gene transfer and genetic duplication for nitrogenase evolution and diversification. Finally, we discuss the importance of H2 synthesis in cyanobacteria, as well as strategies and challenges to improve cyanobacterial H2 production.

Sponges of the genus Leucascus are frequently recognised as possessing anastomosed tubes with choanocytes, and cortical and atrial membranes with pinacocytes. In the last years, five species of other genera were transferred to Leucascus , and several other species were suggested but not formally included in this genus. In the present work, all these species accepted or suggested as Leucascus were revised. According to our results, Leucascus is now composed of nine species: L. clavatus , L. leptoraphis comb. nov., L. lobatus , L. neocaledonicus , L. protogenes comb. nov., L. roseus , L. simplex (type species), L. albus sp. nov., and L. flavus sp. nov. The presence of spines in the apical actine of the tetractines had never been observed in Leucascus , but it was found in all species with tetractines in their skeletons. Some species were transferred from Leucascus to the genus Ascoleucetta , which is revalidated here based on important differences in the cortex. Modifications are also proposed in the definition of both genera. Based on our results, the family Leucascidae is now composed of Ascaltis , Leucascus and Ascoleucetta .

A lógica fuzzy vem sendo aplicada em diversas áreas do conhecimento e nela cada variável possui um conjunto de funções de pertinência representadas em um universo X as quais geralmente exprimem o conjunto de variáveis linguísticas do sistema. Neste trabalho se propõe uma discussão sobre a utilização da lógica fuzzy na aceitação sensorial, a partir da opinião dos provadores quanto ao pão de forma enriquecido com amêndoas e proteínas de faveleira (Cnidosculus phyllacanthus (Mart.) Pax. et K. Hoffm.). Observou-se que o método fuzzy desenvolvido é de fácil utilização e o usuário do sistema só necessita encontrar a curva do centro de gravidade (CG) com maior valor final para indicar a amostra de maior aceitação, podendo utilizar as derivadas das curvas CG no auxílio da detecção da melhor escolha em algumas situações em que lancem dúvida, além de ser utilizado em conjunto com outro método estatístico.Fuzzy logic is applied in many areas of knowledge. In this logic every variable has a set of functions, represented in an X universe, which generally expresses a linguistic variables group of the system. This paper proposes a discussion on the use of fuzzy logic in sensory acceptance, from the taster's views of 'faveleira' (Cnidosculus phyllacanthus (Mart.)Pax. et K. Hoffm.) almonds and protein bread. It was observed that the developed controller method is easy to use and system's user only needs to find the curve of the GC with a final higher value to indicate the sample with greater acceptance. The user may use the derived CG curves aiding in the detection of the best choice in some situations, furthermore used in conjunction with other statistical method.

A lógica fuzzy vem sendo aplicada em diversas áreas do conhecimento e nela cada variável possui um conjunto de funções de pertinência representadas em um universo X as quais geralmente exprimem o conjunto de variáveis linguísticas do sistema. Neste trabalho se propõe uma discussão sobre a utilização da lógica fuzzy na aceitação sensorial, a partir da opinião dos provadores quanto ao pão de forma enriquecido com amêndoas e proteínas de faveleira (Cnidosculus phyllacanthus (Mart.) Pax. et K. Hoffm.). Observou-se que o método fuzzy desenvolvido é de fácil utilização e o usuário do sistema só necessita encontrar a curva do centro de gravidade (CG) com maior valor final para indicar a amostra de maior aceitação, podendo utilizar as derivadas das curvas CG no auxílio da detecção da melhor escolha em algumas situações em que lancem dúvida, além de ser utilizado em conjunto com outro método estatístico. Fuzzy logic is applied in many areas of knowledge. In this logic every variable has a set of functions, represented in an X universe, which generally expresses a linguistic variables group of the system. This paper proposes a discussion on the use of fuzzy logic in sensory acceptance, from the taster's views of 'faveleira' (Cnidosculus phyllacanthus (Mart.)Pax. et K. Hoffm.) almonds and protein bread. It was observed that the developed controller method is easy to use and system's user only needs to find the curve of the GC with a final higher value to indicate the sample with greater acceptance. The user may use the derived CG curves aiding in the detection of the best choice in some situations, furthermore used in conjunction with other statistical method.

The Amazon has a well-defined wet austral summer monsoon and dry winter monsoon precipitation regime and experienced a sequence of drought events in the last 13 years. This study performs a comparative assessment of observed and predicted climate conditions during the three most recent drought events in the Amazon, in 1997–1998, 2004–2005 and 2009–2010, with emphasis on how these events affected the regional monsoon-like precipitation regime. A century long Negro River level time series at Manaus is investigated, applying extreme values theory for estimating return periods of these major drought events. Possible teleconnections of river levels at Manaus and sea surface temperature at remote regions are explored. Large scale oceanic and atmospheric conditions are investigated to highlight the mechanisms associated with the observed drought conditions, particularly during the dry monsoon season. Satellite estimates are used for diagnosing biomass burning aerosol and discuss possible contributions to the observed precipitation deficits in the 2005 and 2010 drought events during the dry monsoon season. The study is concluded with an analysis of the performance of seasonal precipitation predictions for the dry monsoon seasons of July to September 1998, 2005 and 2010 produced with the operational seasonal prediction system used at the Center for Weather Forecasts and Climate Studies (CPTEC) of the Brazilian National Institute for Space Research (INPE). This system was capable of producing 1 month in advance drought warning for the three investigated events, relevant for helping the government and local population make decisions for reducing drought impacts in the Amazon region. Copyright © 2012 Royal Meteorological Society

The aquiferous system is an essential character of poriferans and supports their monophyly. Within the Calcarea, this system displays its greatest variety and traditionally is classified as: asconoid, syconoid, sylleibid, and leuconoid. Species of Leucascus, however, present a different type of aquiferous system composed of anastomosed (interconnected) choanocyte tubes and have an atrium lacking choanoderm. There is such confusion about the classification of the aquiferous system of Leucascus that, depending on the author, it has been classified as asconoid, syconoid, or leuconoid. Therefore, in the present work, we describe a new type of aquiferous system for Leucascus: the solenoid aquiferous system. This new aquiferous system is defined by the presence of anastomosed tubes internally lined by choanocytes and atrium without choanoderm. Although no deep phylogenetic significance has been attributed to the aquiferous system, the solenoid system raises important evolutionary questions about the variety of systems found among the poriferans.

Friedreich's ataxia is the most common inherited ataxia. Ninety-six percent of patients are homozygous for GAA trinucleotide repeat expansions in the first intron of the frataxin gene. The remaining cases are compound heterozygotes for a GAA expansion and a frataxin point mutation. We report here the identification of 10 novel frataxin point mutations, and the detection of a previously described mutation (G130V) in two additional families. Most truncating mutations were in exon 1. All missense mutations were in the last three exons coding for the mature frataxin protein. The clinical features of 25 patients with identified frataxin point mutations were compared with those of 196 patients homozygous for the GAA expansion. A similar phenotype resulted from truncating mutations and from missense mutations in the carboxy-terminal half of mature frataxin, suggesting that they cause a comparable loss of function. In contrast, the only two missense mutations located in the amino-terminal half of mature frataxin (D122Y and G130V) cause an atypical and milder clinical presentation (early-onset spastic gait with slow disease progression, absence of dysarthria, retained or brisk tendon reflexes, and mild or no cerebellar ataxia), suggesting that they only partially affect frataxin function. The incidence of optic disk pallor was higher in compound heterozygotes than in expansion homozygotes, which might correlate with a very low residual level of normal frataxin produced from the expanded allele.

Relatively few calcareous sponges (Porifera, Calcarea) have been recorded from Brazil, mostly due to a reduced number of studies focused exclusively on these generally small, cryptic organisms. To fill in part this gap, the taxonomy and distribution of calcareous sponges from Potiguar Basin (Rio Grande do Norte state, Northeastern Brazil) are studied here. Thirty one specimens were collected by dredging, trawling and SCUBA diving from 2003 to 2007 from depths of 2 to 160 m. Seven species were found and are described here. Two of these species are new to science (Leucetta potiguar sp. nov. and Paraleucilla sphaerica sp. nov.), and four are new records for Rio Grande do Norte state (Clathrina aurea, Leucascus roseus, Leucetta floridana, and Leucandra serrata). An identification key for the species of Calcarea from Potiguar Basin is provided here.

Friedreich's ataxia is the first known autosomal recessive disease caused by an unstable trinucleotide expansion mutation. The most frequent mutation is expansion of a GAA repeat in the first intron of gene X25. We studied transmission of the expanded GAA repeat in 37 Friedreich's ataxia pedigrees and analysed blood and sperm alleles in eight patients. We showed intergenerational instability in 84% of the alleles with an overall excess of contractions. Both contractions and expansions of the GAA repeat occurred in maternal transmission with a stronger tendency to expand for smaller repeats and to contract for longer repeats. Paternally transmitted alleles contracted only. Parental age and the intergenerational change in expansion size were directly correlated in maternal transmission and inversely in paternal transmission. The size of the GAA expansion was slightly lower in patients than heterozygous carriers. Sperm analysis confirmed the tendency to contract of paternal alleles, which was more marked with ageing. The degree of contraction of the GAA repeat in sperm was much higher than that found in intergenerational transmission and was directly related to the repeat size. A blood expanded allele reverted to normal size in the sperm of one patient. This study suggests the existence of different mutational mechanisms in Friedreich's ataxia alleles, which occur both pre- and post-zygotically.

A mamoneira, mesmo tendo sua produtividade diminuída sob déficit hídrico, é considerada resistente a condições de baixa precipitação, apresentando-se como alternativa de geração de renda para o semi-árido. Entre os derivados de sua produção se destaca o ácido graxo ricinoléico utilizado na substituição de derivados de petróleo. A carência de informações sobre os efeitos da escassez de água em regiões nas quais não se consegue satisfazer sua demanda evapotranspirométrica justificou o presente trabalho, em que se estudou o consumo de água e a eficiência do uso da água para duas cultivares (BRs(-1)49 e BRs(-1)88), submetidas a diferentes conteúdos de água no solo. O estudo foi conduzido em uma estufa na UFCG, de 7 de fevereiro a 5 de agosto de 2004, na qual as plantas foram dispostas em esquema fatorial 2 x 4, constituído de duas cultivares e quatro conteúdos de água no solo (40, 60, 80 e 100% de água disponível no solo) com três repetições. Analisando-se fitomassa, o consumo de água e a eficiência do uso pelas plantas, constatou-se que ambas as cultivares mostram comportamento similar ao das plantas conduzidas sem déficit hídrico, apresentando alta produção de fitomassa, o que indica elevada eficiência na transformação da água consumida em matéria seca.Although the productivity of castor bean reduces under water deficit it is considered resistant to low precipitation conditions, thus constituting an alternative form of income for the semi-arid. Among its production derivates the ricinoleic acid stands out as a substitute for petroleum derivates. The scarce information on the effects of water deficit on castor seed production in areas where the low precipitation does not satisfy the evapotranspiration needs motivated this study, which deals with the water consumption and use efficiency of two castor cultivars (BRs(-1)49 and BRs(-1)88) submitted to different soil water contents. The study was conducted in a greenhouse located at the Federal University of Campina Grande - Campus I, during the period of February 7 to August 5, 2004, following a 2 x 4 factorial design with two cultivars, four soil water levels (40, 60, 80 and 100% of the available soil water) and three replicates. Analyzing the phytomass, the water consumption and water use efficiency it was observed that both cultivars show a similar behavior. The plants without water stress, presented a high phytomass production, thus indicating an elevated efficiency on the conversion of water used into dry matter.

Background and purpose: The p38 kinase regulates the release of proinflammatory cytokines including tumour-necrosis factor-alpha (TNF alpha) and is regarded as a potential therapeutic target in rheumatoid arthritis (RA). Using the novel p38 inhibitor Org 48762-0, we investigated the therapeutic potential of p38 inhibition and compared this to anti-mouse (m) TNF alpha antibody treatment in murine collagen-induced arthritis (CIA). Experimental approach: Pharmacological profiles of Org 48762-0 were characterized in kinase assays, cellular assays and in lipopolysaccharide (LPS)-induced inflammation in mice. The effects of Org 48762-0 and of mTNF alpha-neutralization on established arthritis were examined in murine CIA. Key results: Org 48762-0 potently inhibited p38 alpha kinase with a high degree of kinase selectivity. In cellular assays, Org 48762-0 reduced LPS-induced TNF alpha release. Oral administration of Org 48762-0 in mice showed drug-like pharmacokinetic properties and inhibited LPS-induced cytokine production. These pharmacological characteristics of Org 48762-0 prompted a comparison of therapeutic efficacy with mTNF alpha-neutralization in CIA. Org 48762-0 and anti-mTNF alpha antibody treatment equally inhibited development of arthritis when evaluated macroscopically. Radiological analyses revealed protection against bone damage for both treatments, although statistical difference was reached with Org 48762-0 treatment only. Further, micro-computed tomographical and histopathological analyses confirmed the protective effects of Org 48762-0 on joint damage. Conclusions and implications: Pharmacological targeting of p38 kinase provided good protection against joint tissue damage in CIA. In our experiments, neutralization of mTNF alpha produced less prominent suppression of bone damage. Our data suggest a therapeutic potential for selective and potent p38 inhibitors in RA.

Calcareous sponges (Calcarea: Calcinea) are being described, for the first time, from São Paulo State, Brazil. A total of five species are described, two of which are new to science (Clathrina alcatraziensis sp. nov. and Leucascus roseus sp. nov.). The remaining three species have their distribution widened (Clathrina aurea, C. conifera and C. tetractina).

Chondrilla nucula is a morphologically simple species defined by the presence of spherasters in its skeleton. For a long time C. nucula was considered a cosmopolitan species, however, in 1999, a molecular study demonstrated the existence of a species complex with at least four cryptic species, besides C. nucula. At that time, those four new species were not formally described because the morphological differences observed did not reflect species differences and were attributed to plasticity. Since then, researchers working on Chondrilla species suggested that the skeletal organization, the distribution of spherulous cells and the type of surface should be used to differentiate species. Individuals of Chondrilla aff. nucula from one population in Arraial do Cabo, Rio de Janeiro State, Brazil, were collected monthly and analysed in order to test the validity of those three characters to separate the species of Chondrilla. Moreover, to determine if the differences in spicule dimensions previously found were seasonal, we also quantified the diameter of the spherasters throughout the year. Our results demonstrate that all analysed characters are extremely variable and that spicule size can vary even at the same locality. The present study shows the importance of testing the variability of morphological characters before describing new species. Unfortunately, until valid taxonomic characters are found, the Atlantic Chondrilla species will have to remain undescribed.

The cashew ( Anacardium occidentale L.) crop is of great economic and social importance for the Northeast Brazil, a region affected by water and soil salinity. The present study was conducted in a greenhouse to evaluate the sensitivity of the clone CCP76 of the precocious dwarf cashew to the saline stress in the pre-flowering stage. The treatments consisted of five levels of electrical conductivity of the irrigation water (ECw: 0.8; 1.6; 2.4; 3.2 and 4.0 dS m -1 , at 25 oC), delineated in randomized blocks, with three repetitions. Significant effect of the increase of ECw was observed on the plant growth. The precocious dwarf cashew is moderately sensitive to salinity of irrigation water in pre-flowering stage. The value of ECw = 1.6 dS m -1 was considered as a threshold tolerance for the pre-flowering of the studied genotype. The use of saline waters above 0.8 dS m -1 of electrical conductivity produced reduction in water consumption by the plants.

Stomata control the exchange of CO2 and water vapor in land plants. Thus, whereas a constant supply of CO2 is required to maintain adequate rates of photosynthesis, the accompanying water losses must be tightly regulated to prevent dehydration and undesired metabolic changes. Accordingly, the uptake or release of ions and metabolites from guard cells is necessary to achieve normal stomatal function. The AtQUAC1, an R-type anion channel responsible for the release of malate from guard cells, is essential for efficient stomatal closure. Here, we demonstrate that mutant plants lacking AtQUAC1 accumulated higher levels of malate and fumarate. These mutant plants not only display slower stomatal closure in response to increased CO2 concentration and dark but are also characterized by improved mesophyll conductance. These responses were accompanied by increases in both photosynthesis and respiration rates, without affecting the activity of photosynthetic and respiratory enzymes and the expression of other transporter genes in guard cells, which ultimately led to improved growth. Collectively, our results highlight that the transport of organic acids plays a key role in plant cell metabolism and demonstrate that AtQUAC1 reduce diffusive limitations to photosynthesis, which, at least partially, explain the observed increments in growth under well-watered conditions.

Sponges of the genus Leucascus are frequently recognised as possessing anastomosed tubes with choanocytes, and cortical and atrial membranes with pinacocytes. In the last years, five species of other genera were transferred to Leucascus, and several other species were suggested but not formally included in this genus. In the present work, all these species accepted or suggested as Leucascus were revised. According to our results, Leucascus is now composed of nine species: L. clavatus, L. leptoraphis comb. nov., L. lobatus, L. neocaledonicus, L. protogenes comb. nov., L. roseus, L. simplex (type species), L. albus sp. nov., and L.flavus sp. nov. The presence of spines in the apical actine of the tetractines had never been observed in Leucascus, but it was found in all species with tetractines in their skeletons. Some species were transferred from Leucascus to the genus Ascoleucetta, which is revalidated here based on important differences in the cortex. Modifications are also proposed in the definition of both genera. Based on our results, the family Leucascidae is now composed of Ascaltis, Leucascus and Ascoleucetta.

Two new species of the genus Vosmaeropsis are described: V. recruta sp. nov. and V. complanatispinifera sp. nov., both from the Brazilian coast. These species possess arrow-shaped large diactines in their skeletons, which are being described for the first time in Vosmaeropsis. In addition, the latter species presents another remarkable character: the apical actines of its atrial tetractines is flattened and provided with spines. Vosmaeropsis complanastispinifera sp. nov. is being proposed based on the re-analysis of specimens that in the past were assigned to V. sericata. Thus, with the reallocation of these specimens to the newly described species, the current distribution range of V. sericata does not include the Brazilian northeastern coast. Vosmaeropsis now comprises 23 species. Most of them present an endemic pattern of distribution, but the genus is widely distributed in tropical and temperate regions around the world.

Since the earlier anaerobic treatment systems, the design concepts were improved from classic reactors like septic tanks and anaerobic ponds, to modern high rate reactor configurations like anaerobic filters, UASB, EGSB, fixed film fluidized bed and expanded bed reactors, and others. In this paper, anaerobic reactors are evaluated considering the historical evolution and types of wastewaters. The emphasis is on the potential for application in domestic sewage treatment, particularly in regions with a hot climate. Proper design and operation can result in a high capacity and efficiency of organic matter removal using single anaerobic reactors. Performance comparison of anaerobic treatment systems is presented based mostly on a single but practical parameter, the hydraulic retention time. Combined anaerobic reactor systems as well as combined anaerobic and non-anaerobic systems are also presented.

The Intelligent Forecast System (IFS) consists of three units: the main unit is a multilayer neural network; the second unit is based on fuzzy logic, which is used to prepare the data; and the third unit is based on a genetic algorithm and is used to determine the best set of data to be analyzed. The data were set up by using a strategy that classifies the terms of the series into parts with the same quantity of elements. Each part is called a window. The IFS was trained by using a group of windows, which is called a set of training windows. Next, the IFS was applied to solve several problems related to the evaluation of structural integrity. The IFS was applied as an identification strategy and forecast of parameters in a flexible structure. Additionally, by using the IFS we have developed a new strategy to analyze the temporary series obtained from vibrations of flexible structures with the distribution of variable mass. The results obtained in this research, using the IFS applied as an identification strategy and forecast of parameters in a flexible structure, show the effectiveness of the IFS.

Although septic tanks are amply used for on site sewage treatment, these units have serious drawbacks: the removal efficiency of organic material and suspended solids is low, the units are costly and occupy a large area and operational cost is high due to the need for periodic desludging. In this paper an innovative variant of the UASB reactor is proposed as an alternative for the septic tank. This alternative has several important advantages in comparison with the conventional septic tank: (1) Although the volume of the UASB reactor was about 4 times smaller than the septic tank, its effluent quality was superior, even though small sludge particles were present, (2) desludging of the UASB reactor is unnecessary and even counterproductive, as the sludge mass guarantees proper performance, (3) the UASB reactor is easily transportable (compact and light) and therefore can be produced in series, strongly reducing construction costs and (4) since the concentration of colloids in the UASB effluent is much smaller than in the ST effluent, it is expected that the infiltration of the effluent will be much less problematic.

Our study reports on the occurrence of a new species of Leucetta (Calcinea, Calcarea) from the Southern Ocean, Leucetta giribeti, sp. nov., collected in the shallow waters (15m depth) of Deception Island, South Shetland Islands. This new taxon is described based on a combination of morphological and molecular data, including the description of oocytes, embryos, larvae and sperm found in the choanosome. While female reproductive elements showed great similarities with those of other calcineans, sperm is reported here for the first time in the whole Calcinea subclass. Sperm cells are flagellated and possess a typical spermatic mid-piece, which is usually observed in cnidarians. In our phylogenetic analyses, we recovered Leucetta giribeti, sp. nov. as sister species of a clade formed by species of the genera Leucetta, Pericharax and Leucettusa. Although the clade in which Leucetta giribeti, sp. nov. is placed is supported by molecular and morphological features, we cannot propose a new genus due to uncertainties regarding the type species of the genus, Leucetta primigenia Haeckel, 1872. Our study reinforces the relevance of integrative approaches in the description of new taxa and contributes to resolving the poorly known reproductive patterns of Antarctic sponge species.

Several specimens of calcareous sponges were sampled on artificial substrates of a recreational marina in Salvador, Bahia state, on the northeastern coast of Brazil. They belong to four species, two of which are new: Leucandra serrata, Paraleucilla incomposita, Sycon bellum sp. nov., and S. avus sp. nov. Leucandra serrata is recorded for the first time at the coast of Bahia, filling a large gap in the geographical distribution of this species, while the description of the specimens of P. incomposita revealed new characters that had previously been overlooked. Finally, the new species of Sycon described here differ from all the previously known species of the genus due to their skeletal organisation and spicule size. Our results increase the number of species known in Bahia from 13 to 17 and emphasise that areas with a large amount of artificial hard substrates can potentially harbour an interesting and unknown diversity of calcareous sponges.

Amphoriscus is a widespread genus with 17 species. A new species was found in SE Brazil and it represents the second species of this genus in Brazilian waters. Amphoriscus pedunculatus sp. nov. has a special structure, a peduncle, to attach to the substrate. Special attachment structures are not very common in the class Calcarea but this is the third species of the genus with a peduncle. Besides peduncle, another attachment structure found in some species of Amphoriscus is the root-tuft, an attachment structure composed of diactines and anchor-like triactines or tetractines. The evolution of these attachment structures in Amphoriscus is not known but they have also been found out of this genus, suggesting that these structures appeared several times during the evolution of Calcarea or that species currently classified in different genera are in fact congeneric.

Autophagy is a highly conserved cellular mechanism in eukaryotes allowing the degradation of cell constituents. It is of crucial significance in both cellular homeostasis and nutrient recycling. During energy limited conditions plant cells can metabolize alternative respiratory substrates, such as amino acids, providing electrons to the mitochondrial metabolism via the tricarboxylic acid (TCA) cycle or electron transfer flavoprotein/ electron transfer flavoprotein ubiquinone oxidoreductase (ETF/ETFQO) system. Our recent study reveals the importance of autophagy in the supply of amino acids to provide energy through alternative pathways of respiration during carbon starvation. This fact apart, autophagy seems to have more generalized effects related not only to amino acid catabolism but also to metabolism in general. By further comparing the metabolic data obtained with atg mutants with those of mutants involved in the alternative pathways of respiration, we observed clear differences between these mutants, pointing out additional effects of the autophagy deficiency on metabolism of Arabidopsis leaves. Collectively, our data point to an interdependence between mitochondrial metabolism and autophagy and suggest an exquisite regulation of primary metabolism under low energetic conditions.

Among 101 patients homozygous for GAA expansion within the X25 gene, 11 from 8 families had Friedreich’s ataxia with retained reflexes in the lower limbs (FARR). These patients had a lower occurrence of decreased vibration sense, pes cavus, and echocardiographic signs of left ventricular hypertrophy than the 90 FA patients with areflexia. The mean age at onset was significantly later (26.6 ± 11.4 vs. 14.2 ± 6.9 years), and the mean size of the smaller allele was significantly less (408 ± 252 vs. 719 ± 184 GAA triplets) in FARR patients. The neurophysiological findings were consistent with milder peripheral neuropathy and milder impairment of the somatosensory pathways in FARR patients.

We studied the factors that might influence onset age in Friedreich's ataxia in 41 cases (20 male, 21 female) homozygous for GAA expansion on the first intron of X25 gene. Patients came from 18 multiplex families (13 couples, 5 triplets). Mean age (SD) was 18.1 (8.9) years and did not differ by gender. Onset age and the sizes of the smaller (GAA1) and the larger (GAA2) allele in each pair showed high intrafamily correlation. We found an inverse correlation between age at onset and GAA1 size, but not between age at onset and GAA2 size. Stepwise multiple regression of onset age on GAA1 size, sibling onset age, and GAA2 size showed that GAA1 accounts for 73% of onset age variance, and sibling onset age for an additional 13%. The study demonstrates that, in addition to GAA expansion size, other environmental or genetic familial factors influence disease expression.

Three new species of calcareous sponges from the coast of Bahia State, NE Brazil are described. All of them belong to the genus Paraleucilla (Calcaronea, Leucosolenida, Amphoriscidae): P. solangeae sp. nov., P. oca sp. nov., and P. incomposita sp. nov. The number of species recorded from the Bahia coast has thus increased from 10 to 13. Including these new species, there are now 50 calcareous sponge species known from the entire Brazilian coast. Paraleucilla is now composed of 11 species, six of them occurring along the southwestern Atlantic Ocean. The remaining species occur mainly in the Indian Ocean, and also in the Pacific Ocean, Red Sea and Mediterranean Sea. An identification key for all Paraleucilla species is provided. This paper is dedicated to the memory of Professor Solange Peixinho, to acknowledge her contribution to our understanding of the biodiversity of Calcarea from the Bahia coast in Brazil.

Total lipids from the Brazilian brown seaweed Sargassum vulgare were extracted with chloroform/methanol 2:1 and 1:2 (v/v) at room temperature. After performing Folch partition of the crude lipid extract, the lipids recovered from the Folch lower layer were fractionated on a silica gel column eluted with chloroform, acetone and methanol. The fraction eluted with methanol, presented a strong orcinol-positive band characteristic of the presence of sulfatides when examined by TLC. This fraction was then purified by two successive silica gel column chromatography giving rise to fractions F4I86 and F4II90 that exhibited strong activity against herpes simplex virus type 1 and 2. The chemical structures present in both fractions were elucidated by ESI-MS and 1H/13C NMR analysis HSQC fingerprints based on their tandem–MS behavior as Sulfoquinovosyldiacylglycerols (SQDGs). The main SQDG present in both fractions and responsible for the anti-herpes activity observed was identified as 1,2-di-O-palmitoyl-3-O-(6-sulfo-α-d-quinovopyranosyl)-glycerol.

International audience; Friedreich's ataxia (FRDA) is an autosomal recessive, degenerative disease that involves the central and peripheral nervous systems and the heart. A gene, X25, was identified in the critical region for the FRDA locus on chromosome 9q13. This gene encodes a 210-amino acid protein, frataxin, that has homologs in distant species such as Caenorhabditis elegans and yeast. A few FRDA patients were found to have point mutations in X25, but the majority were homozygous for an unstable GAA trinucleotide expansion in the first X25 intron.