Transgenic chloroplasts are efficient sites for high-yield production of the vaccinia virus envelope protein A27L in plant cells.

Orthopoxviruses (OPVs) have recently received increasing attention because of their potential use in bioterrorism and the occurrence of zoonotic OPV outbreaks, highlighting the need for the development of safe and cost-effective vaccines against smallpox and related viruses. In this respect, the production of subunit protein-based vaccines in transgenic plants is an attractive approach. For this purpose, the A27L immunogenic protein of vaccinia virus was expressed in tobacco using stable transformation of the nuclear or plastid genome. The vaccinia virus protein was expressed in the stroma of transplastomic plants in soluble form and accumulated to about 18% of total soluble protein (equivalent to approximately 1.7 mg/g fresh weight). This level of A27L accumulation was 500-fold higher than that in nuclear transformed plants, and did not decline during leaf development. Transplastomic plants showed a partial reduction in growth and were chlorotic, but reached maturity and set fertile seeds. Analysis by immunofluorescence microscopy indicated altered chlorophyll distribution. Chloroplast-synthesized A27L formed oligomers, suggesting correct folding and quaternary structure, and was recognized by serum from a patient recently infected by a zoonotic OPV. Taken together, these results demonstrate that chloroplasts are an attractive production vehicle for the expression of OPV subunit vaccines

Physiological evaluations of tomato cultivars under high temperature stress conditions

Previous studies have demonstrated that the yield reduction of tomato under sub-high temperatures is also related to a decline in the photosynthetic rates. In this work nine tomato landraces were cultivated in two different areas in Campania under three different conditions: open field with normal (April) and late (May) sowing and under plastic tunnel. Photosynthetic parameters, maximum quantum efficiency of PSII, chlorophylls, antioxidants contents and yield were determined. All the landraces showed variable responses for almost all the traits investigated in the different conditions. In the genotypes cultivated under tunnel, where temperatures reached 38-42 °C, there was a clear correlation between some photosynthetic parameters (i.e ETR) and final yield. This correlation was not found in normal and late sown plants. High temperatures affected chlorophylls and antioxidant contents. In particular there was a reduction in the content of chlorophylls and carotenoids and an increase in the level of ascorbic acid when plants were subjected to high temperatures. Genotypes potentially tolerant to high temperatures have been identified through the various parameters investigated

Chamomile essential oils exert anti-inflammatory effects involving human and murine macrophages: Evidence to support a therapeutic action

Ethnopharmacological relevance: Chamomile (M. chamomilla L.) is an herbaceous plant from family Astereaceae, that has a long history of use in traditional medicine. It has been used as herbal remedies for thousands of years to treat several diseases, including infections, neuropsychiatric, respiratory, gastrointestinal, and liver disorders. Chronic inflammation is involved in the pathogenesis of most infectious and non-infectious diseases and macrophages are considered the major cellular players that drive disease initiation and maintenance. Aim of the study: The aim of this study was to evaluate the variation in the chemical profile of the essential oil of M. chamomilla plants collected in three experimental field sites in the Molise region. Additionally, we evaluated the pharmacological mechanism behind the anti-inflammatory effect of M. chamomilla essential oils. Material and methods: Three essential oils (called GC1, GC2 and GC3) were extracted from aerial parts of M. chamomilla by hydrodistillation and chemical composition was analyzed by gas chromatography-mass spectrometry (GC-MS). The essential oils were tested for their ability to modulate pro-inflammatory murine macrophages and human peripheral blood mononuclear cells (PBMCs) functions. Results: The chemical analysis of the samples revealed the presence of a high content of the oxygenated sesquiterpenes that represented more than the half of the entire oils. GC1, GC2 and GC3 essential oils significantly attenuated LPS/IFN-γ-induced inflammation by reducing M1 polarization. In details, they showed significant anti-inflammatory property by inhibiting NO, TNF-α and IL-6 production. These effects were correlated to a suppression of LPS-mediated p65 activation, the critical transactivation subunit for NF-κB transcription factor. Oxidative stress may trigger macrophages activation and elicit strong immune responses. Our study demonstrated that GC1, GC2 and GC3 were highly effective at increasing GCL and HMOX-1 anti-oxidant enzymes expression leading to the rapid scavenging of ROS. The antioxidant activity of these oils was explained throughout the activation of NRF2 signaling pathway. Next, we demonstrated that essential oils were able to reduce CD4+ T cell activation which are also involved in inflammatory processes. Conclusions: Our data describe for the first time that chamomile essential oils exerted their anti-inflammatory and antioxidant activity by modulating macrophages and CD4+ T cells-mediate immune response

Functional ingredients produced by culture of Koliella antarctica

Unicellular algae represent an interesting source of bioactive food ingredients. In this study, fatty acids and carotenoids of Koliella antarctica, a psychrophylic Antarctic unicellular alga, were investigated. K. antarctica was cultivated at different temperatures and harvested at the early exponential growth phase and at the late exponential growth phase. After 240 h, at the end of the late exponential growth phase, the cell biomass concentration was 1.1 g L- 1 at 10 °C while it was 2.2 g L- 1 at 15 °C. The lipid profile of K. antarctica was rich in polyunsaturated fatty acids (PUFA) particularly at the early exponential phase (50.8% of total fatty acids). The comparison of data obtained from algae cultivated at 10 °C and 15 °C, showed at 15 °C a strong increase in saturated lipids and a decrease in PUFA. At the same time the presence of EPA and DHA was marginally affected, with a percentage decrease around 15-20%. The productivity of EPA and DHA was similar at the two growth temperatures with maximum values of 0.6 mg L- 1 day- 1 and 1.3 mg L- 1 day- 1, at 10 and 15 °C, respectively. In K. antarctica cultures, the concentration of lutein and astaxanthin increased during growth up to 1.4 g per 100 g dry weight for both compounds. This is one of the highest concentrations reported in the literature for carotenoid production by unicellular algae, with a total carotenoid productivity of 3.1 mg L- 1 day- 1. Although optimization studies to increase K. antarctica biomass production were not yet carried out, data suggested that this alga is a promising source for low cost production of valuable bioactive compounds. The performance recorded at 15 °C suggested that it could be particularly suitable for outdoor cultivation during the winter season in the Mediterranean region