E3 ubiquitin ligase Cbl-b potentiates the apoptotic action of arsenic trioxide by inhibiting the PI3K/Akt pathway

Arsenic trioxide (ATO) is a strong inducer of apoptosis in malignant hematological cells. Inducible phosphatidyl inositol 3 kinase (PI3K)-Akt activation promotes resistance to ATO. In the present study, we evaluated whether E3 ubiquitin ligase Cbl-b, a negative regulator of PI3K activation, is involved in the action of ATO. The effect of ATO on cell viability was measured by the Trypan blue exclusion assay or by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptosis was determined by flow cytometry and protein expression was assayed by Western blotting. ATO decreased the viability of HL60 cells and induced cellular apoptosis, which was accompanied by transient activation of Akt. The PI3K/Akt inhibitor, LY294002, significantly increased ATO-induced apoptosis (P < 0.05). In addition, ATO up-regulated the expression of Cbl-b proteins. Furthermore, ATO inhibited cell viability with an IC50 of 18.54 μM at 24 h in rat basophilic leukemia-2H3 cells. ATO induced cellular apoptosis with transient activation of Akt and Cbl-b was also up-regulated. Rat basophilic leukemia-2H3 cells transfected with a dominant negative (DN) Cbl-b mutation showed overexpression of Cbl-b (DN) and enhanced Akt activation. Compared with cells transfected with vector, ATO-induced apoptosis was decreased and G2/M phase cells were increased at the same concentration (P < 0.05). The PI3K/Akt inhibitor, LY294002, re-sensitized Cbl-b (DN) overexpressing cells to ATO and reversed G2/M arrest (P < 0.05). Taken together, these results suggest that Cbl-b potentiates the apoptotic action of ATO by inhibition of the PI3K/Akt pathway.

Overexpression of Fc receptor-like 1 associated with B-cell activation during hepatitis B virus infection

The role of B cells in the pathogenesis of hepatitis B virus (HBV) infection has not been explored in depth. In the present study, the activation status of B cells from peripheral blood of healthy controls (N = 20) and patients with acute hepatitis B (AHB, N = 15) or chronic hepatitis B (CHB, N = 30) was evaluated by measuring the expression levels of B-cell activation markers CD69 and CD86, using quantitative real-time PCR and flow cytometry. Moreover, the potential mechanism underlying B-cell activation during HBV infection was further investigated by analyzing the expression profile of FCRL1, an intrinsic activation molecule of B cells. An elevation in the levels of B-cell activation markers including CD69 and CD86 was observed in the AHB patients (44.31 ± 9.27, 27.64 ± 9.26%) compared to CHB patients (30.35 ± 11.27, 18.41 ± 6.56%, P < 0.05), which was still higher than healthy controls (12.23 ± 7.84, 8.22 ± 3.43%, P < 0.05). Furthermore, the expression of FCRL1 was found to be similar to B-cell activation markers, which was highest in AHB patients (70.15 ± 17.11%), lowest in healthy donors (36.32 ± 9.98%, P < 0.05) and half-way between these levels in patients with CHB (55.17 ± 12.03%, P < 0.05). The results were positively associated with aberrant B-cell activation. These data suggest that B cells can play a role in HBV infection, and therefore more effort should be devoted to exploring their functions.

STATISTICAL ANALYSYS OF THE SCFE OF A BRAZILAN MINERAL COAL

The influence of some process variables on the productivity of the fractions (liquid yield times fraction percent) obtained from SCFE of a Brazilian mineral coal using isopropanol and ethanol as primary solvents is analyzed using statistical techniques. A full factorial 23 experimental design was adopted to investigate the effects of process variables (temperature, pressure and cosolvent concentration) on the extraction products. The extracts were analyzed by the Preparative Liquid Chromatography-8 fractions method (PLC-8), a reliable, non destructive solvent fractionation method, especially developed for coal-derived liquids. Empirical statistical modeling was carried out in order to reproduce the experimental data. Correlations obtained were always greater than 0.98. Four specific process criteria were used to allow process optimization. Results obtained show that it is not possible to maximize both extract productivity and purity (through the minimization of heavy fraction content) simultaneously by manipulating the mentioned process variables.

Chemical and biochemical characterization of soybean produced under drought stress

Brazil is the second soybean (Glycine max L. Merrill) producer and exporter in the world. In 2005, soybean cultivated in the southeastern region of the country suffered drought stress imposed by adverse high temperatures and low humidity during its reproductive stage. Little information is available regarding the effect of drought stress on the quality of grains. In this study chemical and biochemical characteristics of five soybean samples belonging to three different cultivars grown under drought stress were evaluated. The samples did not meet standards for marketing and contained high amounts of green seeds. Grains were analyzed for appearance, 100 seed weight, humidity, water activity, proteins, lipids, lipoxygenase 1 activity, peroxides, and pigment contents after harvest and after 20 months of storage at room temperature. Acidity was measured also after 30 months of storage. The values of water activity and humidity were 0.6-0.7 and 8.7-11.9%, respectively, and they did not change during storage time, but there was an increase in acidity, which alludes to lipase activity. The activity of lipoxygenase 1 was greatly affected. Immediately after harvest, the green pigments were represented mainly by pheophytin a, followed by pheophytin b, small quantities of chlorophyll b and chlorophyll a, and traces of other chlorophyll derivatives. After 20 months of storage almost all green pigments had disappeared. Drought stress probably enhanced membrane permeability, which led to a lower pH and promoted transformation of chlorophylls to pheophytins.

Postharvest UV-B treatments increased fructose content of tomato (Solanum lycopersicon L. cv. Tayfun F1) harvested at different ripening stages

Abstract In the postharvest stage, taste and flavor are the key components of the marketability of tomato. Therefore, greater emphasis is now being placed on improving traits such as sugar content. In this study postharvest ultraviolet-B (UV-B) treatments on sugar, total soluble solids, and color of tomatoes harvested at different stages were investigated. Tomatoes harvested at turning, pink, and red stages were treated with two different doses of UV-B irradiation: UVB4 and UVB8. Color L* and hue angle values of tomatoes treated with UV-B were found to be high, which means the red color of tomatoes was improved. UVB4 treatments increased the color a* and saturation index values of tomatoes at pink and red harvest stage, although it did not affect at the turning stage. Additionally, UV-B irradiation treatments had no effect on sucrose content of the tomatoes. Fructose, glucose, and TSS content of tomatoes treated with UVB8 at red harvest stage were found to be high. Hence, the results obtained from this study are of great importance in terms of providing an increase in the amount of sugar without the need for breeding, and also consumer satisfaction.

Hepatitis B vaccination in hemodialysis patients: different points of view for conclusion

The present letter to the editor shows the other aspect of Hepatitis B vaccination in hemodialysis patients.

Tolerance level of green seed in soybean seed lots after storage

The occurrence of green soybean seed due to forced maturation or premature plant death caused by drought or foliar and/or root diseases has been common in several Brazilian production areas. Physiological quality of seed lots with green seed may have their germination and vigor potentials affected and therefore discarded by the grain industry. The objective of this experiment was to determine the maximum tolerated level of green seed in soybean seed lots, which is information of major importance for seed producers when taking the decision whether to sell these lots. Soybean seed of the cultivars CD 206, produced in Ubirata, Parana, and ‘FMT Tucunare,’ produced in Alto Garças, Mato Grosso, were used in the study. Green seed and yellow seed of both cultivars were mixed in the following proportions: 0%, 3%, 6%, 9%, 12%, 15%, 20%, 30%, 40%, 50%, 75% and 100%. Seed quality was evaluated by the germination, accelerated aging, tetrazolium and electrical conductivity tests. The contents of a, b and total chlorophyll in the seed were also determined. A complete randomized block design in a factorial scheme (two cultivars x 12 levels of green seed) was used. Seed quality was negatively affected and chlorophyll contents incremented with the increase in the percentage of green seed. Seed germination, viability and vigor, measured by the accelerated aging test, were not reduced with levels of up to 3% green seed, for both cultivars. Levels above 6% green seed significantly reduced the quality of the seed. The quality of seed lots with 9% or more green seed was significantly reduced to the point that their commercialization is not recommended.

Response of soybean genotypes to the expression of green seed under temperature and water stresses

Soybeans grown under water stress associated with high temperatures during seed maturation and pre-harvest may produce green seed (GS) with expressive reduction in seed quality. The objectives of this study were to evaluate the response of different soybean cultivars grown under these stressful conditions regarding their susceptibility to GS production and to determine the chlorophyll retention levels and the chlorophyllase activity in the seeds. Seeds of four soybean cultivars [BRS 133, CD 206, MG/BR 46 (Conquista) and BRSMG 251 (Robusta)] were grown under greenhouse conditions until R5.5. At R6, the plants were transferred to phytotrons under temperature stress (from 28ºC to 36ºC) and with water stresses of 10% gravimetric moisture, no water and normal supply. Seeds were harvested at R9 when the percentage of GS and weight of 100 seeds were determined. The contents of a, b and total chlorophylls and the chlorophyllase activity were also determined. The expression of GS production under these conditions varied among cultivars: Conquista and Robusta were considered more susceptible to the production of GS compared to 'BRS 133' and 'CD 206'. These cultivars produced lower GS levels, lower chlorophyll retention and higher chlorophyllase activity compared to Robusta and Conquista. Soybean plants submitted to water and temperature stresses produced high levels of GS, which were small, light and had high chlorophyll contents and low chlorophyllase activity. The contents of a, b and total chlorophylls in GS were inversely proportional to the chlorophyllase activity.