Purification and characterization of an exo-polygalacturonase produced by Penicillium viridicatum RFC3 in solid-state fermentation

The pectinolytic enzyme obtained from Penicillium viridicatum RFC by solid-state fermentation was purified to homogeneity by pretreatment with kaolin (40 mg mL(-1) ) and ultrafiltration. followed by chromatography on a Sephadex G50 column. The apparent molecular weight of the enzyme was 24 kDa. Maximal activity occurred at pH 6.0 and at 60 degrees C. The enzyme proved to be an exo-polygalacturonase, releasing galacturonic acid by hydrolysis of highly esterified pectin. The presence of 10 mM Ba2+ increased the enzyme activity by 96% and its thermal stability by 30%. besides increasing its stability at acid pH. The apparent K-m with apple pectin as substrate was 1.82 mg mL(-1) and the V-max was 81 mu mol min(-1). (c) 2007 Elsevier Ltd. All rights reserved.

Label-free DNA detection of hepatitis C virus based on modified conducting polypyrrole films at microelectrodes and atomic force microscopy tip-integrated electrodes

We present a new strategy for the label-free electrochemical detection of DNA hybridization for detecting hepatitis C virus based on electrostatic modulation of the ion-exchange kinetics of a polypyrrole film deposited at microelectrodes. Synthetic single-stranded 18-mer HCV genotype-1-specific probe DNA has been immobilized at a 2,5-bis(2-thienyl)-N-(3-phosphoryl-n-alkyl)pyrrole film established by electropolymerization at the previously formed polypyrrole layer. HCV DNA sequences (244-mer) resulting from the reverse transcriptase-linked polymerase chain reaction amplification of the original viral RNA were monitored by affecting the ion-exchange properties of the polypyrrole film. The performance of this miniaturized DNA sensor system was studied in respect to selectivity, sensitivity, and reproducibility. The limit of detection was determined at 1.82 x 10(-21) mol L-1. Control experiments were performed with cDNA from HCV genotypes 2a/c, 2b, and 3 and did not show any unspecific binding. Additionally, the influence of the spacer length of 2,5-bis(2-thienyl)-N-(3-phosphoryl-n-alkyl)pyrrole on the behavior of the DNA sensor was investigated. This biosensing scheme was finally extended to the electrochemical detection of DNA at submicrometer-sized DNA biosensors integrated into bifunctional atomic force scanning electrochemical microscopy probes. The 18-mer DNA target was again monitored by following the ion-exchange properties of the polypyrrole film. Control experiments were performed with 12-base pair mismatched sequences.

Variabilidade espacial de atributos de um solo sob videira em Vitória Brasil (SP)

Foi pesquisada a variabilidade espacial de alguns atributos físicos e químicos de uma associação de solos cultivada sob videira (Vitis vinifera-L), do município de Vitória Brasil, estado de São Paulo, Brasil. O objetivo foi estudar a dependência espacial de tais atributos, assim como caracterizar as respectivas variabilidades, distribuições de freqüência e números mínimos de subamostras do solo para a cultura da videira. Para isso, coletaram-se dados do solo, dispostos segundo uma malha com 156 pontos amostrais, sendo analisados por meio da geoestatística. As maiores variabilidades foram verificadas para a macroporosidade (MA), P, K, Ca, Mg, SB e CTC, ao passo que as menores foram para a densidade do solo (DS), pH e V. O número mínimo de subamostras, necessário para formar uma amostra composta e representativa, variou de 1 (pH e V) a 241 (Mg), tendo seu valor médio de 64 subamostras. Quanto à dependência espacial, o P e o V apresentaram, respectivamente, forte e fraca dependência, enquanto o restante dos atributos apresentou moderada dependência. Desta forma, o alcance dos atributos físicos variou de 2,56 a 4,32 m, enquanto o dos químicos variou de 1,82 a 5,64 m.

Padronização da determinação da velocidade de condução nervosa sensitiva dos nervos tibial e peroneal de cães clinicamente sadios, pela utilização de eletrodos de superfície

O presente trabalho teve como objetivo a padronização dos valores de referência de velocidade de condução nervosa sensitiva dos nervos tibial e peroneal em cães clinicamente sadios, pela utilização de eletrodos de superfície. em todos os sítios de estimulação, captação, referência e terra foram utilizados eletrodos do tipo jacaré, exceto na captação do estímulo no nervo peroneal, próximo à articulação fêmur-tibial, onde o registro só foi possível com a utilização de eletrodo de agulha. Foram utilizados 30 cães, 11 machos e 19 fêmeas, sem raça definida, com idade entre dois e seis anos. Os valores médios das medidas dos potenciais evocados pela estimulação sensitiva dos nervos tibial e peroneal foram: latência inicial, 1,82±0,30ms (1,30 a 2,55ms) e 1,57±0,29ms (1,01 a 2,16ms), amplitude de pico a pico, 96,48±45,78miV (41,6 a 214miV) e 121,25±57,49miV (54,8 a 299miV) e duração, 1,97±0,69ms (1,01 a 3,56ms) e 2,37±0,85ms (1,11 a 3,94ms), respectivamente. Os valores médios das medidas de velocidade de condução nervosa sensitiva dos nervos tibial e peroneal foram, respectivamente, 62,14+7,71ms (50,0 a 77,2ms) e 65,18+6,42ms (53,8 a 79,2ms), respectivamente.

Debris perturbed by radiation pressure: relative velocities across circular orbits

In the present work we consider a dynamical system of mum size particles around the Earth subject to the effects of radiation pressure. Our main goal is to study the evolution of its relative velocity with respect to the co-planar circular orbits that it crosses. The particles were initially in a circular geostationary orbit, and the particles size were in the range between 1 and 100 mum. The radiation pressure produces variations in its eccentricity, resulting in a change in its orbital velocity. The results indicated the maximum linear momentum and kinetic energy increases as the particle size increases. For a particle of 1 mum the kinetic energy is approximately 1.56 x 10(-7) J and the momentum is 6.27 x 10(-11) kg m/s and for 100 mum the energy is approximately 1.82 x 10(-4) J and the momentum is 2.14 x 10(-6) kg m/s. (C) 2004 COSPAR. Published by Elsevier Ltd. All rights reserved.

Mutagenic activity of sweepings and pigments from a household-wax factory assayed with Salmonella typhimurium

The mutagenic activity of garbage originating from a household wax industry was determined by the Salmonella/microsome assay, using the bacterial strains TA100, TA98 and YG1024. The garbage was obtained by sweeping the floor of the factory at the end of the work shift. Organic compounds were extracted by ultrasound for 30 min in dichloromethane or 70% ethanol. After evaporation of solvent, these extracts (HFS: household-wax factory sweepings) were dissolved in DMSO, and were tested for the mutagenic activity at varying concentrations (HFS-ET: 0.08-0.68 mg/plate, HFS-DCM: 0.60-7.31 mg/plate). The colouring agents (pigments) used in the production of the wax were also dissolved in DMSO and tested with the assay. The concentrations tested for each pigment were: Amaranth: 0.46-3.65 mg/plate, Auramine: 0.15-1.2 mg/plate, Tartrazine: 0.46-3.65 mg/plate and Rhodamine B: 0.22-1.82 mg/plate. Both ET and DCM organic extracts had mutagenic activity, especially in the YG1024 strain. The pigments behaved in a similar way, demonstrating that YG1024 was the most sensitive strain for the detection of mutagenicity, and that metabolization increased the activity. Human exposure (occupational and non-occupational) to industrial residues generated during the household-wax manufacturing and packaging process should be monitored, since this type of garbage is normally deposited in the environment without any control. (C) 2004 Elsevier Ltd. All rights reserved.

Cytotaxonomic analysis of Brazilian species of the genus Amazona (Psittacidae, Aves) and confirmation of the genus Salvatoria (Ribeiro, 1920)

The karyotypes of 12 species of Psittacidae of the genus Amazona were studied: A. aestiva, A. amazonica, A. brasiliensis, A. autumnalis, A. farinosa, A, festiva, A. kawalli, A. ochrocephala, A. pretrei, A. rhodocorytha, A. vinacea and A. xanthops. The metaphases were obtained using a short term culture of leather pulp. Eleven of the twelve analyzed species were karyotypically homogeneous, with only a few divergences in chromosomes 2 and 3. The species A. xanthops showed large karyotypic differences compared to the genus Amazona. Consequently, the genus Salvatoria (Ribeiro, Rev. Mus. Paul. 12: 1-82, 1920) was confirmed, and A. xanthops renamed Salvatoria xanthops. The study showed the chromosomic conservation of the genus Amazona and the need for further taxonomic studies of the karyotype of the Psittacidae.

Root growth and cotton nutrition as affected by liming and soil compaction

Soil columns were produced by filling PVC tubes with a Dark Red Latosol (Acrortox, 22% of clay). A compacted layer was established at the depth of 15 cm in the columns. In the compacted layer, soil was packed to 1.13, 1.32, 1.48, and 1.82 Mg kg(-1), resulting in cone resistances of 0.18, 0.43, 1.20, and 2.50 MPa. Cotton was cropped for 30 days. Lime was applied to raise base saturation to 40, 52, and 67%. The highest base saturation caused a decrease in phosphorus (P) and zinc (Zn) concentrations in the plants. A decrease in root dry matter, length and surface area was also observed. This could be a consequence of lime induced Zn deficiency. Root growth was decreased in the compacted layer, and complete inhibition was noticed at 2.50 MPa. Once the roots got through the compacted layer, there was a growth recovery in the bottom layer of the pots. The increase in base saturation up 52% was effective in preventing a decrease in cotton root length at soil resistances to 1.20 MPa. Where the roots were shorter, there was an increase in nutrient uptake per unit of root surface area, which kept the plants well nourished, except for P.

The azomethine-H modified colorimetric method of boron determination at fertilizers.

The present work studied the azomethine-H colorimetric method for boron determination to fertilizers analysis applications. The reagent azomethine-H needs lights conditions of reaction that, jointed to the big sensibility and specificity, diffused its use in boron's dosage in many materials. The most suitable experimental parameters were established for such colorimetry, and analysis conditions: standard curve between 0,200 e 2,25 mg. L(-1) boron, in maximum absorption at 415nm, I cm cell and reading between 30 and 90 minutes standing after the final homogenization of the boron's solution in analyse and with dye reactive (azomethine If 0,90 % m/v solution at buffer ammonium acetate 1,82 mol . L(-1) - potassium acetate 0,10 mol . L(-1) - acetic acid 1,67 mol . L(-1)-EDTA 2,7 . 10(-2) mol . L(-1)-NTA 2,1.10(-2) mol . L(-1) pH 5,5). Statistics analysis of the results didn't present big differences when the same results were got by the colorimetric method offered and by the potentiometric, from AOAG, whatever in manufactured fertilizers or in lab sintetized fertilizers.

EFFECT OF CHOLINERGIC STIMULATION OF THE AMYGDALOID COMPLEX ON WATER AND SALT INTAKE

The effect of carbachol (80 nmol/mul) injection into the amygdaloid nuclear complex (AMG) on sodium appetite and water intake was studied in male Holtzman rats weighing 240-270 g. Twenty-five satiated rats and 38 water-deprived rats were used in the experiment on water intake. In the experiment on sodium intake, 19 rats were injected with atropine + carbachol and 9 rats with hexamethonium + carbachol. After carbachol injection into the AMG, water intake decreased in rats submitted to 30 h of water deprivation (10.28 +/- 1.04 ml/120 min vs 0.69 +/- 0.22 ml/120 min). The decrease in water intake was blocked by prior local injection of a tropine (20 nmol/1 mul)(11.66 +/- 1.46 ml/120 min vs 0.69 +/- 0.22 ml/120 min), but not of hexamethonium (30 nmol/1 mul), into the AMG. In water-deprived animals, carbachol injection into the AMG caused a decrease in sodium chloride intake (6.16 +/- 1.82 ml/h vs 0.88 +/- 0.54 ml/h) which was blocked by previous injection of hexamethonium but not of a tropine. These results suggest that the cholinergic system of the AMG plays a role in the control of water and salt intake.

A linearly tunable low-voltage CMOS transconductor with improved common-mode stability and its application to (gm)-C filters

A linearly tunable low-voltage CMOS transconductor featuring a new adaptative-bias mechanism that considerably improves the stability of the processed-signal common,mode voltage over the tuning range, critical for very-low voltage applications, is introduced. It embeds a feedback loop that holds input devices on triode region while boosting the output resistance. Analysis of the integrator frequency response gives an insight into the location of secondary poles and zeros as function of design parameters. A third-order low-pass Cauer filter employing the proposed transconductor was designed and integrated on a 0.8-mum n-well CMOS standard process. For a 1.8-V supply, filter characterization revealed f(p) = 0.93 MHz, f(s) = 1.82 MHz, A(min) = 44.08, dB, and A(max) = 0.64 dB at nominal tuning. Mined by a de voltage V-TUNE, the filter bandwidth was linearly adjusted at a rate of 11.48 kHz/mV over nearly one frequency decade. A maximum 13-mV deviation on the common-mode voltage at the filter output was measured over the interval 25 mV less than or equal to V-TUNE less than or equal to 200 mV. For V-out = 300 mV(pp) and V-TUNE = 100 mV, THD was -55.4 dB. Noise spectral density was 0.84 muV/Hz(1/2) @1 kHz and S/N = 41 dB @ V-out = 300 mV(pp) and 1-MHz bandwidth. Idle power consumption was 1.73 mW @V-TUNE = 100 mV. A tradeoff between dynamic range, bandwidth, power consumption, and chip area has then been achieved.

Eficiência de absorção e utilização de potãssio por plantas de soja em solução nutritiva

A greenhouse experiment was conducted in a factorial design of randomized complete blocks, with three replications, to study plantage response of soybean cultivars on efficiency: update (EAB); conversion (EC) and utilization, as well as utilization index (IU) by soybean cultivars. Four soybean cultivars (IAC 17, IAC 18, IAC 11 e FT 2) were grown in nutrient solutions with either adequate or deficient K levels (0.5 and 1.82 mmol. L-1) and harvested 20, 40 and 60 days after plant emergence (DAPE). The plants were separated in top, median and lower leaves, stems and roots, dried, ground and analyzed for K contents. Regardless of cultivar, in deficient plants K was translocated from the other organs to median leaves, in order to maintain its K contents. This behavior was particularly evident in 'IAC 11' and 'IAC 18'. Well nourished plants showed higher EAB than deficient plants. 'IAC 17' and 'IAC 18' showed higher EAB at either adequate or deficient K. levels. 'FT 2' showed highest EUK in deficient solutions at 40 DAPE (immediately before flowering), and the lowest EUK at 60 DAPE. The utilization index was helpful in determining cultivar plasticity to adapt to K deficiency. The highest plasticity was observed for 'FT 2' at 40 DAPE and for 'IAC 18' at 60 DAPE. K-deficient plants were more efficient in EC, yielding more dry matter for K uptake unit.

Determination of vanadium in urine by electrothermal atomic absorption spectrometry using hot injection and preconcentration into the graphite tube

In this work it was developed a procedure for the determination of vanadium in urine samples by electrothermal atomic absorption spectrometry using successive injections for preconcentration into a preheated graphite tube. Three 60 μL volumes were sequentially injected into the atomizer preheated to a temperature of 110°C. Drying and pyrolysis steps were carried out after each injection. A chemical modifier, barium difluoride (100 mg L-1), and a surfactant, Triton X-100 (0.3% v v-1), were added to the urine sample. When injecting into a hot graphite tube, the sample flow-rate was 0.5 μL s-1. The limits of detection and quantification were 0.54 and 1.82 without preconcentration, and 0.11 and 0.37 μg L-1 with preconcentration, respectively. The accuracy of the procedure was evaluated by an addition-recovery experiment employing urine samples. Recoveries varied from 96.0 to 103% for additions ranging from 0.8 to 3.5 μg L-1 V. The developed procedure allows the determination of vanadium in urine without any sample pretreatment and with minimal dilution of the sample.

Susceptibilidade in vitro a antimicrobianos da Mannheimia haemolytica e da Pasteurella multocida isoladas de ovinos sadios e com doenças respiratórias

Pasteurella multocida and Mannheimia haemolytica (P. haemolytica) are associated with ovine respiratory diseases. With the purpose of observe the susceptibility in vitro of these organisms against antimicrobials, were collected samples of nasopharingeal (n=180) and oropharingeal (n=82) from ovines healthy and with respiratory disease. Among the antimicrobials tested, the sensibility was greater for enrofloxacin (100%) and florfenicol (100%), for both bacteria. The greater resistance indices for M. haemolytica and P. multocida were observed with tetracyclin (15.64% and 17.65% respectively) and penicillin (1.82% and 4.2%).

Search for resonant t̄t production in lepton+jets events in pp collisions at √s=7 TeV

A model-independent search for the production of heavy resonances decaying into top-antitop quark pairs is presented. The search is based on events containing one lepton (muon or electron) and at least two jets selected from data samples corresponding to an integrated luminosity of 4.4-5.0 fb -1 collected in pp collisions at √s =7 TeV. Results are presented from the combination of two dedicated searches optimized for boosted production and production at threshold. No excess of events is observed over the expected yield from the standard model processes. Topcolor Z′ bosons with narrow (wide) width are excluded at 95% confidence level for masses below 1.49 (2.04) TeV and an upper limit of 0.3 (1.3) pb or lower is set on the production cross section times branching fraction for resonance masses above 1 TeV. Kaluza-Klein excitations of a gluon with masses below 1.82 TeV (at 95% confidence level) in the Randall-Sundrum model are also excluded, and an upper limit of 0.7 pb or lower is set on the production cross section times branching fraction for resonance masses above 1 TeV.[Figure not available: See fulltext.] © 2012 CERN for the benefit of the CMS collaboration.