Switchable Solvent Selective Extraction of Hydrophobic Antioxidants from Synechococcus bigranulatus

Hydrophobic molecules, in particular, carotenoids, have been directly extracted from Synechococcus bigranulatus ACUF680 by means of secondary amine switchable solvent N-ethylbutylamine (EBA) without any other pretreatment. EBA was able to extract hydrophobic molecules from both fresh and frozen biomass at the same extent of the conventional procedure (about 20% of dry biomass). In particular, selective extraction of a zeaxanthin-enriched fraction (green fraction, GF) and a β-carotene-enriched fraction (orange fraction, OF) was obtained. The ratio between zeaxanthin and β-carotene was 4.4 ± 1.5 for GF, 0.07 ± 0.06 for OF, and about 1 for conventional extraction. These fractions showed in vitro antioxidant activity (IC50 values of 0.056 ± 0.013 and 0.024 ± 0.008 mg mL-1 for GF and OF, respectively) and biocompatibility on immortalized cells. Moreover, OF and GF were able to protect cells from oxidative stress, both before and after thermal treatment. Results clearly indicate that EBA is a good candidate to specifically extract β-carotene and zeaxanthin from the wet biomass of S. bigranulatus without affecting their biological activity. Skipping energy-intensive operations to break the cells and using either fresh or frozen biomass may be the driving factors to use EBA switchable solvent on an industrial scale

Identification of Quantitative Trait Loci for Flowering Time in a Field-Grown \u3cem\u3eLolium Perene x Lolium Multiflorum\u3c/em\u3e Mapping Population

Perennial ryegrass (L. perenne) and annual, or Italian, ryegrass (L. multiflorum) are considered to be separate species by the seed trade, and are used and bred for distinct purposes. However, the two species are cross-fertile. Seed producers rely on the different flowering times of the two species to produce pure seed. Flowering times can overlap, leading to genetic mixing. Contamination of perennial ryegrass seed lots with annual types is an expensive problem for grass seed producers in western Oregon, USA

Evaluation of the Antiviral Response to Zanamivir Administered Intravenously for Treatment of Critically Ill Patients With Pandemic Influenza A (H1N1) Infection

A retrospective nationwide study on the use of intravenous (IV) zanamivir in patients receiving intensive care who were pretreated with oseltamivir in the Netherlands was performed. In 6 of 13 patients with a sustained reduction of the viral load, the median time to start IV zanamivir was 9 days (range, 4–11 days) compared with 14 days (range, 6–21 days) in 7 patients without viral load reduction (P = .052). Viral load response did not influence mortality. We conclude that IV zanamivir as late add-on therapy has limited effectiveness. The effect of an immediate start with IV zanamivir monotherapy or in combination with other drugs need to be evaluated

CD4+ and CD8+ T Cells Can Act Separately in Tumour Rejection after Immunization with Murine Pneumotropic Virus Chimeric Her2/neu Virus-Like Particles

BACKGROUND: Immunization with murine pneumotropic virus virus-like particles carrying Her2/neu (Her2MPtVLPs) prevents tumour outgrowth in mice when given prophylactically, and therapeutically if combined with the adjuvant CpG. We investigated which components of the immune system are involved in tumour rejection, and whether long-term immunological memory can be obtained. METHODOLOGY AND RESULTS: During the effector phase in BALB/c mice, only depletion of CD4+ and CD8+ in combination, with or without NK cells, completely abrogated tumour protection. Depletion of single CD4+, CD8+ or NK cell populations only had minor effects. During the immunization/induction phase, combined depletion of CD4+ and CD8+ cells abolished protection, while depletion of each individual subset had no or negligible effect. When tumour rejection was studied in knock-out mice with a C57Bl/6 background, protection was lost in CD4-/-CD8-/- and CD4-/-, but not in CD8-/- mice. In contrast, when normal C57Bl/6 mice were depleted of different cell types, protection was lost irrespective of whether only CD4+, only CD8+, or CD4+ and CD8+ cells in combination were eradicated. No anti-Her2/neu antibodies were detected but a Her2/neu-specific IFNgamma response was seen. Studies of long-term memory showed that BALB/c mice could be protected against tumour development when immunized together with CpG as long as ten weeks before challenge. CONCLUSION: Her2MPtVLP immunization is efficient in stimulating several compartments of the immune system, and induces an efficient immune response including long-term memory. In addition, when depleting mice of isolated cellular compartments, tumour protection is not as efficiently abolished as when depleting several immune compartments together

High efficiency of alphaviral gene transfer in combination with 5-fluorouracil in a mouse mammary tumor model

Copyright: Copyright 2014 Elsevier B.V., All rights reserved.Background: The combination of virotherapy and chemotherapy may enable efficient tumor regression that would be unachievable using either therapy alone. In this study, we investigated the efficiency of transgene delivery and the cytotoxic effects of alphaviral vector in combination with 5-fluorouracil (5-FU) in a mouse mammary tumor model (4 T1).Methods: Replication-deficient Semliki Forest virus (SFV) vectors carrying genes encoding fluorescent proteins were used to infect 4 T1 cell cultures treated with different doses of 5-FU. The efficiency of infection was monitored via fluorescence microscopy and quantified by fluorometry. The cytotoxicity of the combined treatment with 5-FU and alphaviral vector was measured using an MTT-based cell viability assay. In vivo experiments were performed in a subcutaneous 4 T1 mouse mammary tumor model with different 5-FU doses and an SFV vector encoding firefly luciferase.Results: Infection of 4 T1 cells with SFV prior to 5-FU treatment did not produce a synergistic anti-proliferative effect. An alternative treatment strategy, in which 5-FU was used prior to virus infection, strongly inhibited SFV expression. Nevertheless, in vivo experiments showed a significant enhancement in SFV-driven transgene (luciferase) expression upon intratumoral and intraperitoneal vector administration in 4 T1 tumor-bearing mice pretreated with 5-FU: here, we observed a positive correlation between 5-FU dose and the level of luciferase expression.Conclusions: Although 5-FU inhibited SFV-mediated transgene expression in 4 T1 cells in vitro, application of the drug in a mouse model revealed a significant enhancement of intratumoral transgene synthesis compared with 5-FU untreated mice. These results may have implications for efficient transgene delivery and the development of potent cancer treatment strategies using alphaviral vectors and 5-FU.publishersversionPeer reviewe

Gender differences in the use of cardiovascular interventions in HIV-positive persons; the D:A:D Study

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