Production of the baculovirus-expressed dengue virus glycoprotein NS1 can be improved dramatically with optimised regimes for fed-batch cultures and the addition of the insect moulting hormone, 20-Hydroxyecdysone

A perennial problem in recombinant protein expression is low yield of the product of interest. A strategy which has been shown to increase the production of baculovirus-expressed proteins is to utilise fed-batch cultures. One disadvantage of this approach is the time-consuming task of optimising the feeding strategy. Previously, a statistical optimisation routine was applied to develop a feeding strategy that increased the yield of beta-Galactosidase (beta-Gal) by 2.4-fold (Biotechnol. Bioeng, 59 (1998) 178). This involves the single addition of nutrient concentrates (amino acids, lipids. glucose and yeastolate ultrafiltrate) into Sf9 cell cultures grown in SF900II medium. In this study, it is demonstrated that this optimised fed-batch strategy developed for a high-yielding intracellular product beta-Gal could be applied successfully to a relatively low-yielding glycosylated and secreted product such as the dengue virus glycoprotein NS1. Optimised batch infections yielded 4 mug/ml of NS1 at a peak cell density of 4.2 x 10(6) cells/ml. In contrast. optimised fed-batch infections exhibited a 3-fold improvement in yield, with 12 mug ml of NS1 produced at a peak cell density of 11.3 x 10(6) cells/ml. No further improvements in yield were recorded when the feed volumes were doubled and the peak cell density was increased to 23 x 10(6) cells/ml, unless the cultures were stimulated by the addition of 4 mug/ml of 20-Hydroxyecdysone (an insect moulting hormone). In this case, the NS1 yield was increased to 20 mug/ml. which was nearly 5-fold higher than optimised batch cultures. (C) 2002 Elsevier Science B.V. All rights reserved.

Improved nitrogen removal by application of new nitrogen-cycle bacteria

In order to meet increasingly stringent European discharge standards, new applications and control strategies for the sustainable removal of ammonia from wastewater have to beimplemented. In this paper we discuss anitrogen removal system based on the processesof partial nitrification and anoxic ammoniaoxidation (anammox). The anammox process offers great opportunities to remove ammonia in fully autotrophic systems with biomass retention. No organic carbon is needed in such nitrogenremoval system, since ammonia is used a selectron donor for nitrite reduction. The nitrite can be produced from ammonia in oxygen-limited biofilm systems or in continuous processes without biomass retention. For successful implementation of the combined processes, accurate biosensors for measuring ammonia and nitrite concentrations, insight inthe complex microbial communities involved, and new control strategies have to be developed and evaluated.

Identification of a digalactosyl ononitol from seeds of adzuki bean (Vigna angularis)

A digalactosyl ononitol was isolated from seeds of adzuki bean (Vigna angularis [Willd.] Ohwi et Ohasi). Analysis of hydrolysis products and NMR spectroscopy established its structure as O-alpha-D-galactopyranosyl-(1-->6)-O-alpha-D-galactopyranosyl-(1-->3)-O-methyl- D-myo-inositol. (C) 2003 Elsevier Ltd. All rights reserved.

Actively shielded multi-layer gradient coil designs with improved cooling properties

In standard cylindrical gradient coils consisting of a single layer of wires, a limiting factor in achieving very large magnetic field gradients is the rapid increase in coil resistance with efficiency. This is a particular problem in small-bore scanners, such as those used for MR microscopy. By adopting a multi-layer design in which the coil wires are allowed to spread out into multiple layers wound at increasing radii, a more favourable scaling of resistance with efficiency is achieved, thus allowing the design of more powerful gradient coils with acceptable resistance values. Previously this approach has been applied to the design of unshielded, longitudinal, and transverse gradient coils. Here, the multi-layer approach has been extended to allow the design of actively shielded multi-layer gradient coils, and also to produce coils exhibiting enhanced cooling characteristics. An iterative approach to modelling the steady-state temperature distribution within the coil has also been developed. Results indicate that a good level of screening can be achieved in multi-layer coils, that small versions of such coils can yield higher efficiencies at fixed resistance than conventional two-layer (primary and screen) coils, and that performance improves as the number of layers of increases. Simulations show that by optimising multi-layer coils for cooling it is possible to achieve significantly higher gradient strengths at a fixed maximum operating temperature. A four-layer coil of 8 mm inner diameter has been constructed and used to test the steady-state temperature model. (C) 2003 Elsevier Inc. All rights reserved.

The influence of light spectra, UV-A, and growth regulators on the in vitro seed germination of Senecio cineraria DC.

This study was carried out to investigate the effects of light spectra, additional UV-A, and different growth regulators on the in vitro germination of Senecio cineraria DC. Seeds were surface-sterilized and inoculated in MS medium to evaluate the following light spectra: white, white plus UV-A, blue, green, red or darkness. The maximum germinability was obtained using MS0 medium under white light (30%) and MS + 0.3 mg L-1 GA3 in the absence of light (30.5%). S. cineraria seeds were indifferent to light. Blue and green lights inhibited germination. Different concentrations of gibberellic acid (GA3) (0.1; 0.4; 0.6; 0.8; 1.0 and 2.0 mg L-1) and indole-3-acetic acid IAA (0.1; 0.3 and 1.0 mg L-1) were evaluated under white light and darkness. No concentration of GA3 enhanced seed germination percentage under white light. However, when the seeds were maintained in darkness, GA3 improved germination responses in all tested concentrations, except at 1.0 mg L-1. Under white light, these concentrations also increased the germination time and reduced germination rate. Germination rate, under light or darkness, was lower using IAA compared with GA3.

Drying kinetics of jatropha seeds

Given the necessity of developing jatropha cultivation equipment, this work adjusted different mathematical models to experimental data obtained from the drying of jatropha seeds submitted to different drying conditions and selected the best model to describe the drying process. The experiment was carried out at the Federal Institute of Goiás - Rio Verde Campus. Seeds with initial moisture content of approximately 0.50 (kg water/kg dry matter) were dried in a forced air-ventilated oven, at temperatures of 45, 60, 75, 90 and 105°C to moisture content of 0.10 ± 0.005 (kg water/kg dry matter). The experimental data were adjusted to 11 mathematical models to represent the drying process of agricultural products. The models were compared using the coefficient of determination, chi-square test, relative mean error, estimated mean error and residual distribution. It was found that the increase in the air temperature caused a reduction in the drying time of seeds. The models Midilli and Two Terms were suitable to represent the drying process of Jatropha seeds and between them the use of the Midili model is recommended due to its greater simplicity.

Effect of acetic acid on rice seeds coated with rice husk ash

Flooded rice cultivation promotes anaerobic conditions, favoring the formation of short chain organic acids such as acetic acid, which may be toxic to the crop. The objective of this study was to evaluate the effect of acetic acid on rice seeds coated with rice husk ash. The experiment was arranged in a 2 x 5 x 5 factorial randomized design, with two cultivars (IRGA 424 and BRS Querência), five doses of coating material (0, 2, 3,4 e 5 g kg-1 seed) and five concentrations of acetic acid (0, 3, 6, 9 and 12 mM), with 4 replications, totaling 50 treatments. The variables first count of germination, germination, shoot and root length, dry weight of shoots and roots were recorded. The results showed that coating rice seeds with rice husk ash up to 5 g kg-1 seed does not influence the performance of rice seeds of cultivars IRGA 424 and BRS Querência when exposed to concentrations of 12 mM acetic acid. The presence of acetic acid in the substrates used for seed germination reduced the vigor and viability of seeds of cultivars IRGA 424 and BRS Querência, as well as seedling development, affecting mainly the roots of BRS Querência.

Bioactive albumin functionalized polylactic acid membranes for improved biocompatibility

Biocompatibility is a major challenge for successful application of many biomaterials. In this study the ability to coat chemically and enzymatically activated poly(L-lactic acid) (PLA) membranes with heat denatured human serum albumin to improve biocompatibility was investigated. PLA membranes hydrolyzed with NaOH or cutinase and then treated with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, hydrochloride (EDAC) as a heterobifunctional cross-linker promoted the coupling single bondCOOH groups on PLA membranes and single bondNH2 groups of heat denatured human serum albumin. This resulted in increased hydrophilicity (lowest water contact angles of 43° and 35°) and highest antioxidant activity (quenching of 79 μM and 115 μM tetramethylazobisquinone (TMAMQ) for NaOH and cutinase pretreated membranes, respectively). FTIR analysis of modified PLA membranes showed new peaks attributed to human serum albumin (amide bond, NH2 and side chain stretching) appearing within 3600–3000 cm−1 and 1700–1500 cm−1 (Fig. 3). MTT studies also showed that osteoblasts-like and MC-3T3-E1 cells viability increased 2.4 times as compared to untreated PLA membranes. The study therefore shows that this strategy of modifying the surfaces of PLA polymers could significantly improve biocompatibility.

Kinetics of solute leachate from imbibing Caesalpinia echinata Lam. (Brazilwood) seeds

The electrical conductivity of leachates from imbibing seeds has been used as a vigor test for several species. The adaptation of this methodology to different species requires knowledge on the leaching kinetics of electrolytes. For Brazilwood seeds, the classic method was not satisfactory and rapid tests are essential because they have low storage capacity at room temperature. Leaching kinetics during seed imbibition is a function of physiological quality, presence or absence of seed coat, imbibing temperature and the initial moisture content of seed. In this study, the electrolyte leaching rate of six different categories of seeds, from two regions, was evaluated in seeds with and without seed coat and incubated with different moisture contents and at different temperatures. The results showed that the electrolyte leaching rate in Brazilwood seeds is independent of the physiological quality, the presence or absence of seed coat and imbibition temperature, but these factors changed the total amount of electrolytes leached. The leaching rate increased in the first few minutes of imbibition, suggesting that the adjustment of the methodology must consider the reduction in imbibition time, reduction in temperature, use of a controlled and slower pre-imbibition, and replacement of the imbibition solution after the first few minutes.

Chemical treatment of papaya seeds aiming at long-term storage and control of damping off

Damping off is a nursery disease of great economic importance in papaya and seed treatment may be an effective measure to control. The aim of this work was to evaluate the quality of papaya seeds treated with fungicides and stored under two environmental and packaging conditions. Additionally, the efficiency of fungicide treatments in the control of damping-off caused by Rhizoctonia solani was evaluated. Papaya seeds were treated with the fungicides Captan, Tolylfluanid and the mixture Tolylfluanid + Captan (all commercial wettable powder formulations). Seeds of the control group were not treated. The seeds were stored for nine months in two conditions: packed in aluminum coated paper and kept at 7 ± 1ºC and in permeable kraft paper and kept in non-controlled environment. At the beginning of the storage and every three months the seed quality (germination and vigor tests), emergence rate index, height, dry mass and damping of plants in pre and post-emergence (in contaminated substrate and mycelia-free substrate) were analyzed. Both storage conditions as well as the fungicide treatments preserved the germination and seed vigor. In the infested substrate, seedling emergence was favored by fungicides, but in post-emergence, fungicides alone did not control the damping off caused by R. solani. Symptoms of damping off were not observed in the clean substrate. The results showed that the fungicide treatments may be used to pretreat papaya seed for long-term storage and commercialization.

Influence of foliar fertilization with manganese on germination, vigor and storage time of RR soybean seeds

ABSTRACTThis study aimed to evaluate the influence of foliar fertilizer doses containing Mn of phenological stages of suggested application in RR soybeans, to recover management damages with glyphosate at postemergence application on seed vigor in post-harvest and post six months storage. The seeds originated from a field experiment conducted , which included two applications of glyphosate, concomitant with foliar fertilizer in growth stages V4 and V6, with 0.00, 113.50 and 227.00 mg ha-1doses of Mn2+. Germination, GSI (Germination Speed Index), electrical conductivity tests and the first count of seeds were conducted. The application of Mn did not affect the physiological quality of RR soy in postharvest. However, in post-storage, higher doses of Mn had a negative effect on tests of abnormal seedlings, GSI and electrical conductivity. The applications of Mn, regardless of the developmental stage, did not interfere in the germination and first count tests, with and without storage. The electrical conductivity test showed a higher correlation with the seed germination test in the post-harvest treatment.

Initial performance of corn in response to treatment of seeds with humic acids isolated from bokashi

ABSTRACT The humified organic matter presents bioactivity similar to the auxinic effect. As bokashi is produced by a special process of humification, information is needed about the bioactive potential of its humic acids. The objective of this work was studying the initial performance of corn-indicator plants in response to the application of different concentrations of humic acids isolated from bokashi. The corn seeds were treated for 16 hours with solutions containing 0, 10, 20, 30, 40 and 80 mmol L-1 of C in the form of humic acids. Then, the seeds were planted in pots of 1 dm3 containing corrected and fertilized soil, in greenhouse. Growth characteristics of shoot and root systems were evaluated. The results showed that the humic acids extracted from bokashi had positive effects on the initial performance of corn.