Utilization of microsatellites for the analysis of genomic alterations in colorectal cancers in Brazil

Two different pathogenetic mechanisms are proposed for colorectal cancers. One, the so-called "classic pathway", is the most common and depends on multiple additive mutational events (germline and/or somatic) in tumor suppressor genes and oncogenes, frequently involving chromosomal deletions in key genomic regions. Methodologically this pathway is recognizable by the phenomenon of loss of heterozygosity. On the other hand, the "mutator pathway" depends on early mutational loss of the mismatch repair system (germline and/or somatic) leading to accelerated accumulation of gene mutations in critical target genes and progression to malignancy. Methodologically this second pathway is recognizable by the phenomenon of microsatellite instability. The distinction between these pathways seems to be more than academic since there is evidence that the tumors emerging from the mutator pathway have a better prognosis. We report here a very simple methodology based on a set of tri-, tetra- and pentanucleotide repeat microsatellites allowing the simultaneous study of microsatellite instability and loss of heterozygosity which could allocate 70% of the colorectal tumors to the classic or the mutator pathway. The ease of execution of the methodology makes it suitable for routine clinical typing

Secretory TAT-peptide-mediated protein transduction of LIF receptor α-chain distal cytoplasmic motifs into human myeloid HL-60 cells

The distal cytoplasmic motifs of leukemia inhibitory factor receptor α-chain (LIFRα-CT3) can independently induce intracellular myeloid differentiation in acute myeloid leukemia (AML) cells by gene transfection; however, there are significant limitations in the potential clinical use of these motifs due to liposome-derived genetic modifications. To produce a potentially therapeutic LIFRα-CT3 with cell-permeable activity, we constructed a eukaryotic expression pcDNA3.0-TAT-CT3-cMyc plasmid with a signal peptide (ss) inserted into the N-terminal that codes for an ss-TAT-CT3-cMyc fusion protein. The stable transfection of Chinese hamster ovary (CHO) cells via this vector and subsequent selection by Geneticin resulted in cell lines that express and secrete TAT-CT3-cMyc. The spent medium of pcDNA3.0-TAT-CT3-cMyc-transfected CHO cells could be purified using a cMyc-epitope-tag agarose affinity chromatography column and could be detected via SDS-PAGE, with antibodies against cMyc-tag. The direct administration of TAT-CT3-cMyc to HL-60 cell culture media caused the enrichment of CT3-cMyc in the cytoplasm and nucleus within 30 min and led to a significant reduction of viable cells (P < 0.05) 8 h after exposure. The advantages of using this mammalian expression system include the ease of generating TAT fusion proteins that are adequately transcripted and the potential for a sustained production of such proteins in vitro for future AML therapy.

Effects of peeling methods on the quality of cubiu fruits

Cubiu (Solanum sessiliflorum Dunal) is an Amazonian Basin native fruit. Its importance comes from its high contents of pectin. Currently, processing technologies are necessary for the substitution of the traditional system (small crops and small-scale processing) for a larger scale system and thus increase the use of biodiversity and promote the implementation of Local Productive Arrangements of agribusiness in the Amazon. This research aims to evaluate the methods of peeling cubiu. Ripe fruits were divided into lots (150 each) and subjected to the following treatments: immersion in 2.5% NaOH boiling solution for 5 minutes, exposure to water vapor, and immersion in water at 96 ºC for 5, 10, 15 and 20 minutes. The peel released during heat treatment and immediately removed under running tap water. In the control treatment, the fruits were manually peeled (unheated) with a stainless steel knife. The treatments were evaluated for completeness and ease of peeling, tissue integrity, texture, and peroxidase activity. The immersion in 2.5% NaOH boiling solution (5 minutes) stood out as the best treatment since it inhibited the enzymatic browning and intensified the natural yellow color of the cubiu fruit and easily and fully peeled the whole fruit more rapidly without damaging its tissues. This treatment was chosen as the most advantageous because it can promote simultaneous peeling and bleaching. Therefore, it is recommended for cubiu industrial processing.

Physicochemical properties, modifications and applications of starches from different botanical sources

Present trends towards technologies and processes that increase the use of residues make starchy vegetal biomass an important alternative material in various applications due to starch’s versatility, low cost and ease of use when its physicochemical properties are altered. Starch is increasingly used in many industrial applications and as a renewable energy resource. Starch can be modified to enhance its positive attributes and eliminate deficiencies in its native characteristics. In this article, the state of knowledge on conventional and unconventional starches and their properties, characteristics, modifications and applications are reviewed.

Boron extraction and vertical mobility in Paraná State oxisol, Brazil

The deficiency or excess of micronutrients has been determined by analyses of soil and plant tissue. In Brazil, the lack of studies that would define and standardize extraction and determination methods, as well as lack of correlation and calibration studies, makes it difficult to establish limits of concentration classes for analysis interpretation and fertilizer recommendations for crops. A specific extractor for soil analysis is sometimes chosen due to the ease of use in the laboratory and not in view of its efficiency in determining a bioavailable nutrient. The objectives of this study were to: (a) evaluate B concentrations in the soil as related to the fertilizer rate, soil depth and extractor; (b) verify the nutrient movement in the soil profile; (c) evaluate efficiency of Hot Water, Mehlich-1 and Mehlich-3 as available B extractors, using sunflower as test plant. The experimental design consisted of complete randomized blocks with four replications and treatments of five B rates (0, 2, 4, 6, and 8 kg ha-1) applied to the soil surface and evaluated at six depths (0-0.05, 0.05-0.10, 0.10-0.15, 0.15-0.20, 0.20-0.30, and 0.30-0.40 m). Boron concentrations in the soil extracted by Hot Water, Mehlich-1 and Mehlich-3 extractors increased linearly in relation to B rates at all depths evaluated, indicating B mobility in the profile. The extractors had different B extraction capacities, but were all efficient to evaluate bioavailability of the nutrient to sunflower. Mehlich-1 and Mehlich-3 can therefore be used to analyze B as well as Hot Water.

Polarizabilidade atômica efetiva alfad pode ditar a ionização de radioligantes 99Tc m - diaminoditiol alquilamínicos?

Polarizability correlates well with organic ion stabilization in solution and can be defined as a measure of the relative ease of the distortion of the electronic cloud of a dipolar system exposed to an external electric field. The effective atomic polarizability, alphad, has a fundamental influence on chemical reactivity in the gas phase and in solution. In terms of chemical reactivity the charge is generated within the molecule as a positive charge due to protonation, ionization or resulting from the attack of a nucleophilic anion. In this paper, lipoidal diaminedithiol (DADT) perfusion radioligands based on 99Tc m and possessing an alkylamine side chain have been used to check the influence of alphad on their brain uptake. Some new DADT derivatives, respectively DADT-DIPA (diaminedithiol - diisopropylamine), DADT DIBA (diaminedithiol diisobutylamine), DADT-PR (diaminedithiol - branched pyperidine), have been designed to have high nitrogen alkylamine alphad values. In spite of the fact of higher alphad values having been correlated to higher brain uptakes, there isn't a clear mechanism able to trap these radioligands into the brain space.

Emprego da fluorescência de raios X por dispersão de energia para o mapeamento de troncos de árvores em Campinas, São Paulo

Dendrochemistry is based on the determination of elements retained in tree rings, which can be useful to characterize environmental occurrences. This work shows elemental mappings obtained by EDXRF of the cross section of a tree stem. The tree is originated from a polluted area in Campinas, São Paulo. Some profiles, as the sulfur one, show variations that can be attributed to external contributions, whereas the silicon one can be inferred to a protection defense natural mechanism. Besides being simultaneous and multielementar, the main advantages here are its performance through a non-destructive sample treatment and the ease of operation of XRF equipments.

Celdas solares fotoelectroquímicas basadas en Bi2WO6

In this study, photoelectrochemical solar cells based on bismuth tungstate electrodes were evaluated. Bi2WO6 was synthesized by a hydrothermal method and characterized by scanning electron microscopy, UV-Vis reflectance spectroscopy, and X-ray powder diffraction. For comparison, solar cells based on TiO2 semiconductor electrodes were evaluated. Photoelectrochemical response of Grätzel-type solar cells based on these semiconductors and their corresponding sensitization with two inexpensive phthalocyanines dyes were determined. Bi2WO6-based solar cells presented higher values of photocurrent and efficiency than those obtained with TiO2 electrodes, even without sensitization. These results portray solar cells based on Bi2WO6 as promising devices for solar energy conversion owing to lower cost of production and ease of acquisition.

KIT COLORIMÉTRICO PARA DETECÇÃO DE METANOL EM ETANOL COMBUSTÍVEL PARA O MONITORAMENTO DA QUALIDADE DE COMBUSTÍVEIS

A colorimetric kit for methanol detection in ethanol-containing fuels and ease of use in the field was developed and tested. The analysis can detect the presence of methanol in fuels when exceeding specification (0.5% v/v) in about 20 min and its simple instrumentation does not require a specialist. The kit method was successfully validated at gas stations located in São Paulo State and the Federal District.

TEMPERATURE AND SUBSTRATE INFLUENCES ON SEED GERMINATION AND SEEDLING FORMATION IN Callisthene fasciculata Mart. (VOCHYSIACEAE) IN THE LABORATORY1

ABSTRACTCallisthene fasciculata Mart. is a tree belonging to the Vochysiaceae family. Its wood is moderately heavy and resistant and used to make poles, beams, and other structures. The aim of this work was to evaluate seed germination and the initial growth of seedlings of C. fasciculata at different temperatures and in different substrates. Seeds were collected from fruits in the Pantanal de Miranda, Mato Grosso do Sul state, Brazil. In one experiment, the seeds were subjected to constant temperatures of 20, 25, 30 and 35 °C and to alternating temperatures of 20-30 and 25-35 °C (on paper substrate). In another experiment, the seeds were subjected to temperatures of 20 and 25 °C on three substrates (sand, vermiculite and between paper) in a germinator. The experiment had a randomized design, with four replicates of 25 seeds per treatment. The F-values obtained for germination indicated no significant effect of substrate or temperature on the final germination percentage. The analyses revealed no effect of a treatment interaction (temperature x substrate) on either germination or average germination time; however, a treatment interaction effect was observed on the germination speed index. The treatment combinations yielding the best performance were between paper substrate at 20 °C and sand substrate at 25 °C. There was a significant effect of the interaction between temperature and substrate on seedling growth, with increased root growth observed in the between paper substrate at 25 °C and increased aerial component growth in both sand at 20 °C and vermiculite at 25 °C. The between paper treatment at 25 °C yielded the greatest final seedling size. Between paper is the most recommended substrate for the production of seedlings due to its ease of handling and lower probability of contamination.

A forced running wheel system with a microcontroller that provides high-intensity exercise training in an animal ischemic stroke model

We developed a forced non-electric-shock running wheel (FNESRW) system that provides rats with high-intensity exercise training using automatic exercise training patterns that are controlled by a microcontroller. The proposed system successfully makes a breakthrough in the traditional motorized running wheel to allow rats to perform high-intensity training and to enable comparisons with the treadmill at the same exercise intensity without any electric shock. A polyvinyl chloride runway with a rough rubber surface was coated on the periphery of the wheel so as to permit automatic acceleration training, and which allowed the rats to run consistently at high speeds (30 m/min for 1 h). An animal ischemic stroke model was used to validate the proposed system. FNESRW, treadmill, control, and sham groups were studied. The FNESRW and treadmill groups underwent 3 weeks of endurance running training. After 3 weeks, the experiments of middle cerebral artery occlusion, the modified neurological severity score (mNSS), an inclined plane test, and triphenyltetrazolium chloride were performed to evaluate the effectiveness of the proposed platform. The proposed platform showed that enhancement of motor function, mNSS, and infarct volumes was significantly stronger in the FNESRW group than the control group (P<0.05) and similar to the treadmill group. The experimental data demonstrated that the proposed platform can be applied to test the benefit of exercise-preconditioning-induced neuroprotection using the animal stroke model. Additional advantages of the FNESRW system include stand-alone capability, independence of subjective human adjustment, and ease of use.