Cell cycle synchronization and BrdU incorporation as a tool to study the possible selective elimination of ErbB1 gene in the micronuclei in A549 cells

Lung cancer often exhibits molecular changes, such as the overexpression of the ErbB1 gene that encodes epidermal growth factor receptor (EGFR). ErbB1 amplification and mutation are associated with tumor aggressiveness and low response to therapy. The aim of the present study was to design a schedule to synchronize the cell cycle of A549 cell line (a non-small cell lung cancer) and to analyze the possible association between the micronuclei (MNs) and the extrusion of ErbB1 gene extra-copies. After double blocking, by the process of fetal bovine serum deprivation and vincristine treatment, MNs formation was monitored with 5-bromo-2-deoxyuridine (BrdU) incorporation, which is an S-phase marker. Statistical analyses allowed us to infer that MNs may arise both in mitosis as well as in interphase. The MNs were able to replicate their DNA and this process seemed to be non-synchronous with the main cell nuclei. The presence of ErbB1 gene in the MNs was evaluated by fluorescent in situ hybridization (FISH). ErbB1 sequences were detected in the MNs, but a relation between the MNs formation and extrusion of amplified ErbB1could not be established. The present study sought to elucidate the meaning of MNs formation and its association with the elimination of oncogenes or other amplified sequences from the tumor cells.

Effect of enzymatic hydrolysis on some physicochemical properties of root and tuber granular starches

Enzymatic hydrolysis of granular starch is an important tool to provide information about granule structure. Cassava, sweet potato, Peruvian carrot, and potato starches were hydrolyzed by bacterial α-amylase at 37 °C for 48 hours, and the physicochemical properties of the residues from hydrolysis were determined. Cassava starch was the most susceptible to enzyme displaying 20.9% of hydrolysis, whereas potato starch was the most resistant with 5.9%. The granule average size varied from 10.8 to 23.4 μm for Peruvian carrot and potato starches, respectively. With the use of SEM, a smooth granule surface was observed for all native starches. Cassava and sweet potato starches displayed an A-type X-ray diffraction pattern, while Peruvian carrot and potato starches showed a B-type pattern. After hydrolysis, cassava, sweet potato, and Peruvian carrot starches showed some well degraded granules, whereas potato starch presented a slight sign of degradation. The amylose content of the starches decreased with hydrolysis for cassava, sweet potato, and Peruvian carrot starches and was kept unchanged for the potato starch. As expected, intrinsic viscosity and pasting properties decreased for all hydrolyzed starches. There is no difference between thermal properties of native and hydrolyzed starches. These results suggested that hydrolysis occurred in amorphous and crystalline areas of the granules. The B type diffraction pattern in conjunction with the big granule size of the potato starch may have contributed to the greatest resistance of this starch to hydrolysis.

Chemical pattern of brazilian apples: a chemometric approach based on the Fuji and Gala varieties

The chemical composition of apple juices may be used to discriminate between the varieties for consumption and those for raw material. Fuji and Gala have a chemical pattern that can be used for this classification. Multivariate methods correlate independent continuous chemical descriptors with the categorical apple variety. Three main descriptors of apple juice were selected: malic acid, total reducing sugar and total phenolic compounds. A chemometric approach, employing PCA and SIMCA, was used to classify apple juice samples. PCA was performed with 24 juices from Fuji and Gala, and SIMCA, with 15 juices. The exploratory and predictive models recognized 88% and 64%, respectively, as belonging to a mixed domain. The apple juice from commercial fruits shows a pattern related to cv. Fuji and Gala with boundaries from 0.18 to 0.389 g.100 mL-1 (malic acid), from 8.65 to 15.18 g.100 mL-1 (total reducing sugar) and from 100 to 400 mg.L-1 (total phenolic compounds), but such boundaries were slightly shorter in the remaining set of commercial apple juices, specifically from 0.16 to 0.36 g.100 mL-1, from 9.25 to 15.5 g.100 mL-1 and from 180 to 606 mg.L-1 for acidity, reducing sugar and phenolic compounds, respectively, representing the acid, sweet and bitter tastes.

Ripening pattern of guava cv. Pedro Sato

Guava is a fruit with high respiration rates and a very short shelf life. Since information on its respiration pattern is contradictory, the objective was to study the changes occurring in the fruit during ripening and to relate them to the respiration behavior of this fruit. Guavas were picked at the half-ripe stage and stored for 8 days at 22 ± 1 ºC and 78 ± 1% relative humidity. The analyses conducted were: peel and pulp coloration, firmness, total soluble solids (TSS), total titratable acidity (TTA), and ethylene production. According to the results, it was verified that the parameters analyzed apparently do not coincide and are ethylene-independent. There was an accentuated ethylene production during ripening, starting from the 4th day. The ethylene synthesis continued increasing up to the 8th day, when the fruits were already decomposing. It was observed that the firmness decreased sharply in the first three days of ripening, and the skin and pulp color changed during ripening. The TSS, total soluble solids, and the TTA, total titratable acidity, practically did not change during the ripening, even with the increased ethylene production. It can be concluded that guava is a fruit that presents characteristics of climacteric and non-climacteric fruits.

Automated static image analysis as a novel tool in describing the physical properties of dietary fiber

Abstract The growing interest in the usage of dietary fiber in food has caused the need to provide precise tools for describing its physical properties. This research examined two dietary fibers from oats and beets, respectively, in variable particle sizes. The application of automated static image analysis for describing the hydration properties and particle size distribution of dietary fiber was analyzed. Conventional tests for water holding capacity (WHC) were conducted. The particles were measured at two points: dry and after water soaking. The most significant water holding capacity (7.00 g water/g solid) was achieved by the smaller sized oat fiber. Conversely, the water holding capacity was highest (4.20 g water/g solid) in larger sized beet fiber. There was evidence for water absorption increasing with a decrease in particle size in regards to the same fiber source. Very strong correlations were drawn between particle shape parameters, such as fiber length, straightness, width and hydration properties measured conventionally. The regression analysis provided the opportunity to estimate whether the automated static image analysis method could be an efficient tool in describing the hydration properties of dietary fiber. The application of the method was validated using mathematical model which was verified in comparison to conventional WHC measurement results.