Development of two potential diagnostic monoclonal antibodies against human cytomegalovirus glycoprotein B

Human cytomegalovirus glycoprotein B (gB) represents a target for diagnosis and treatment in view of the role it plays in virus entry and spread. Nevertheless, to our knowledge, rare detection of a gB antigen has been reported in transplant patients and limited information is available about diagnostic gB monoclonal antibodies (mAbs). Our aim was to develop gB mAbs with diagnostic potential. Hydrophilic gB peptides (ST: amino acids 27-40, SH: amino acids 81-94) of favorable immunogenicity were synthesized and used to immunize BALB/c mice. Two mAbs, named ZJU-FH6 and ZJU-FE6, were generated by the hybridoma technique and limited serial dilution and then characterized by indirect ELISA, Western blotting, immunoprecipitation, and immunohistochemical staining. The mAbs displayed high titers of specific binding affinities for the ST and SH synthetic peptides at an mAb dilution of 1:60,000 and 1:240,000, respectively. Western blotting and immunoprecipitation indicated that these mAbs recognized both denatured and native gB of the Towne and AD169 strains. The mAbs, when used as the primary antibody, showed positive staining in cells infected with both Towne and AD169 strains. The mAbs were then tested on patients submitted to allogeneic hematopoietic stem cell transplantation. The gB antigen positivity rates of the patients tested using ZJU-FH6 and ZJU-FE6 were 62.0 and 63.0%, respectively. The gB antigen showed a significant correlation with the level of pp65 antigen in peripheral blood leukocytes. In conclusion, two potential diagnostic gB mAbs were developed and were shown to be capable of recognizing gB in peripheral blood leukocytes in a reliable manner.

Effect of an edible crosslinked coating and two types of packaging on antioxidant capacity of castilla blackberries

In order to increase the shelf life and maintain the quality and stability of the biological compounds with antioxidant activity present in Castilla blackberry fruits, a sodium alginate-based edible crosslinked coating was applied, and the fruits were packed in two different plastic containers and stored under refrigeration (3 ± 1 °C). Total antioxidant capacity and its relationship to physicochemical variables such as pH, Brix, and acidity were evaluated in six treatments: uncoated blackberry stored in a macroperforated container (T1) and thermosealed container (T2), without crosslinked coating in a macroperforated container (T3) and thermosealed container (T4), with crosslinked coating (calcium ions) packed in macroperforated container (T5) and thermosealed container (T6). The results indicated that factors such as gas permeability in the coatings, the packaging used, and physicochemical parameters significantly affected the fruit total antioxidant capacity, with the highest level in T1 (0.22 µgEAA/ml) at the end of the essay, which is related to the lowest levels of pH and direct exposure to air. On the other hand, the lowest value was obtained in T6 (0.16 µgEAA/ml) due to the crosslinked coating, packaging in the thermosealed container, and higher pH value. Variations in acidity, Brix, and pH indicate the presence of degenerative processes in the crosslinked coating treatments, which limited the physicochemical changes.

Optimization of microwave drying conditions of two banana varieties using response surface methodology

AbstractOptimization of microwave drying conditions of Luvhele and Mabonde banana varieties were studied using response surface methodology. The drying was performed using a central composite rotatable design for two variables: microwave power level (100, 200 and 300 W) and drying time (40, 26, and 12 min.) for Luvhele; (100, 200 and 300 W) and (42, 27, and 12 min) for Mabonde. The colour and texture (hardness) data were analyzed using ANOVA and regression analysis. The fitness of the models obtained was good as the lack of fit for each of the models was not significant. The coefficient of determination R2 of the models was relatively high, hence the models obtained for the responses were adequate and acceptable. Drying conditions of 178.76 W, 12 min. drying time were found optimum for product quality at a desirability of 0.91 for Luvhele; while 127.67 W, 12 min. with a desirability of 0.86 was predicted for Mabonde. The result of this study could be used as a standard for microwave processing of Luvhele and Mabondebanana varieties.

Influence of the moisture at harvest and drying process of the grains on the level of carotenoids in maize (Zea mays)

AbstractMaize is considered a source of carotenoids; however, these compounds are highly unstable, degraded by high temperatures, exposure to light and presence of oxygen. The objective of this work was to evaluate the influence of the moisture and type of drying applied to grains on the level of carotenoids in yellow maize. The experiment was conducted in a completely randomized design (2 × 4 factorial), two levels of initial moisture at the harvest (22 and 19%) and three types of drying (in the sun; in the shade and in a dryer) and control (no drying). The samples of grains after drying with 12% of final moisture were analyzed by concentration of total carotenoids, carotenes (α-carotene + β-carotene), monohydroxilated carotenoids (β-cryptoxanthin), and xanthophylls (lutein + zeaxanthin). Initial moisture, type of drying and the interaction between moisture versus drying influence (p≤0.05) the levels of carotenoids in grains. This is the first report about the drying conditions and harvest’s initial moisture as influence on the profile and content of carotenoids in maize grains. Based on the results, this work suggested that the harvest be carried out preferably when the grains present 22% humidity, with drying in a dryer or in shade for further use or storage.

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.