Cinnamic Esters of Acyclovir-Synthesis and Biological Activity

In the present study we have synthesized esters of acyclovir (2–4) with cinnamic acids (p-coumaric, ferulic, and sinapic acids) and evaluated them for their antiviral and antioxidant potential. The antiviral activity of the newly synthesized compounds has been tested against human herpes virus 1 (HSV-1) in vitro. The results indicate that none of the synthesized compounds inhibits the tested virus strain. The antioxidant properties have been studied using 2,2-diphenyl-1-picrylhydrazyl (DPPH)* test.     

Genetic engineering of Plum pox virus resistance: ‘HoneySweet’ plum—from concept to product

Sharka disease, caused by Plum pox virus (PPV) was first recorded in Bulgaria during the early twentieth century and since that first report, the disease has progressively spread throughout Europe and more recently to Asia, Africa, North and South America. Few PPV resistance genes have been found to naturally occur in Prunus and this has led to the investigation of biotech approaches to the development of resistance through genetic engineering (GE). A notable example of the utility of this approach is ‘HoneySweet’ plum. PPV protection in this case is based on RNA interference (RNAi) and resistance has been shown to be highly effective, stable, durable, and heritable as a dominant trait. Extensive testing and risk assessment of ‘HoneySweet’ in laboratory, greenhouse and in the field for over 20 years has demonstrated not only the effectiveness but also the safety of the technology. ‘HoneySweet’ has been cleared for cultivation in the USA. By the appropriate regulatory agencies. The development and regulatory approval of ‘HoneySweet’ demonstrate the ability of RNAi technology to contribute to the sustainability of stone fruit production in regions impacted by PPV. Although it has taken almost 100 years since the identification of sharka, we are now able to effectively protect stone fruit species against this disease through the application of GE.     

Isolation and characterization of trehalose tetraester biosurfactants from a soil strain Micrococcus luteus BN56

The bacterium Micrococcus luteus BN56, isolated from soil, was found to produce glycolipid biosurfactants when grown on n-hexadecane as the sole carbon source. The purified glycolipids were characterized using ¹H, ¹³C, ¹H COSY NMR-spectroscopy and ESI-MS spectrometry analyses. The two main products were identified as trehalose tetraesters with molecular mass of 876 and 848 g mol^?1. The purified products reduced the surface tension of water from 72 to 24.1 mN m^?1 and the interfacial tension between water and hexadecane from 43.0 to 1.7 mN m^?1. The CMC of these biosurfactants was found to be 25 mg l^?1. The strain formed stable emulsions with hydrocarbon substrates and was suggested that the hydrophobic cells acted as emulsion-stabilizing agents. The results demonstrate that the strain M. luteus BN56 may be well suited for bioremediation of oil-contaminated environments.     

Molecular cloning and characterisation of a Rab-binding GDP-dissociation inhibitor from Medicago truncatula.

We have isolated and sequenced the full-length cDNA of a GDP-dissociation inhibitor (GDI) from the model legume Medicago truncatula L. The cDNA (MtGDI) contains an open reading frame of 1335 bp, coding for a protein of 444 amino acids with a calculated molecular mass of 49,785 kDa. The deduced amino acid sequence shows significant homology to other plant GDIs, the highest homology being found to GDI from the legume Cicer arietinum (96% identity). The MtGDI was expressed as a N-terminal FLAG-fusion protein in Escherichia coli BL21 (DE3). Its direct interaction with a small G protein of Rab type from Medicago sativa, MsRab11f, was demonstrated in vitro by co-immunoprecipitation using a peptide-specific antibody raised against MtGDI. The dissociation constant of the MtGDI-MsRab11f complex (4 muM) was determined by a surface plasmon resonance (SPR) assay. Real-time RT-PCR and Western blot analyses suggested that MtGDI is ubiquitously expressed in M. truncatula. High levels of MtGDI mRNA were detected in uninfected roots, leaves and root nodules. In etiolated seedlings and cell cultures, the amount of MtGDI mRNA was much lower. In all tissues tested, the peptide-specific anti-MtGDI antibody detected the expected 50 kDa protein in the total protein extracts. MtGDI was found in the cytosol; however, a significant fraction was associated with the intracellular membranes in seedlings and roots indicating a membrane localisation of the protein. A second immunoreactive band was detected in leaves suggesting that more than one GDI isoform exist in M. truncatula.     

The Effect of Dietary Selenium Source and Level on Hen Production and Egg Selenium Concentration

A 16-week experiment was conducted to compare effects of various levels of sodium selenite (SS) and Se-enriched yeast (SY), on the whole-egg Se content and hen’s productivity. One hundred Shaver 579 hens, 27 weeks old, were placed on one of five experimental treatments. Each treatment was replicated four times with five hens per cage. Treatments consisted of feeding a low Se diet without supplementation (basal diet) or basal diet with one of two levels of supplemented Se (0.4 or 0.8 mg/kg) supplied by SS or SY. All supplemented treatments had significantly higher whole-egg Se concentration from basal diet (P < 0.05). On the same supplemented level, hens fed on SY had higher egg Se content from hens feed on SS (P < 0.001). No effects of dietary treatments on egg weight, percentages of dirty and cracked egg, and feed intake and conversion of feed were observed throughout the trial (P < 0.05). In the first 8 weeks, there was no significant difference (P < 0.05) in hen-day egg production among treatments. From the ninth week on to the end of the trial, supplementation of SY to hen’s diet resulted in a higher egg production than SS (P < 0.01).     

Synthesis,antioxidative and antiviral activity of hydroxycinnamic acid amides of thiazole containing amino acid

The synthesis and the biological (antioxidant and antiviral) activities of novel hydroxycinnamic acid amides of a thiazole containing TFA.valine-4-carboxylic acid ethyl ester are reported. The amides have been synthesized from p-coumaric, ferulic and sinapic acids with the corresponding TFA.valine-thiazole-4-carboxylic acid ethyl ester using the coupling reagent N-ethyl-N′-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and 4-(dimethylamino) pyridine (DMAP) as a catalyst. The antioxidant properties of the newly synthesized amides have been studied for then antioxidative activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH)* test. The newly synthesized compounds have been tested against the replication in vitro of influenza virus A (H3N2) and human herpes virus 1 and 2 (HSV-1 and HSV-2).     

Identification and expression analyses of poly [I:C]-stimulated genes in channel catfish (Ictalurus punctatus)

Channel catfish (Ictalurus punctatus) have proven to be an excellent model with which to study immune responses of lower vertebrates. Identification of anti-viral antibodies and cytotoxic cells, as well as both type I and II interferon (IFN), demonstrates that catfish likely mount a vigorous anti-viral immune response. In this report, we focus on other elements of the anti-viral response, and identify more than two dozen genes that are induced following treatment of catfish cells with poly [I:C]. We showed that poly [I:C] induced type I interferon within 2 h of treatment, and that characteristic interferon stimulated genes (ISGs) appeared 6–12 h after exposure. Among the ISGs detected by RT-PCR assay were homologs of ISG15, Mx1, IFN regulatory factor 1 (IRF-1), inhibitor of apoptosis protein-1 (IAP-1) and the chemokine CXCL10. Microarray analyses showed that 13 and 24 cellular genes, respectively, were upregulated in poly [I:C]-treated B cell and fibroblast cultures. Although many of these genes were novel and did not fit the profile of mammalian ISGs, there were several (ISG-15, ubiquitin-conjugating enzyme E2G1, integrin-linked kinase, and clathrin-associated protein 47) that were identified as ISGs in mammalian systems. Taken together, these results suggest that dsRNA, either directly or through the prior induction of IFN, upregulates catfish gene products that function individually and/or collectively to inhibit virus replication.     

The biliprotein C-phycocyanin modulates the early radiation response: A pilot study

This study analyzed the effects of biliprotein C-phycocyanin (C-PC) on the enzymatic antioxidant defence system in lymphocytes of nuclear power-plant workers and non-exposed controls. Changes in the protein levels of manganese super oxide dismutase (MnSOD), catalase and glutathione-S-transferase (GST) were used as markers for early biological effects of a single in vitro exposure of cells to: (i) 2 Gy gamma rays; (ii) 5 μM C-PC; and (iii) a combination of C-PC plus irradiation with 2 Gy. The results showed that C-PC selectively stimulated the lymphocyte antioxidant defence system of occupationally exposed subjects. The activation of the antioxidant protective mechanisms as part of the early radiation response was probably related to the chronic low-dose occupational exposure. The modulating capacity of C-PC at the molecular level may be of interest for the protection of occupationally exposed persons.     

Nitrile degradation by free and immobilized cells of the thermophile Bacillus sp. UG-5B, isolated from polluted industrial waters

A moderately thermophilic Gram-positive, sporulating, rod-shaped strain of Bacillus with nitrile-degrading activity was isolated from polluted industrial waters. Whole cells and cell-free extracts from the end of exponential growth phase expressed 7.6 nkat mg⁻¹ and 2.0 nkat mg⁻¹ benzonitrile-degrading activity, respectively, after cultivation in a fermentor with complex medium containing benzonitrile as an inducer. The benzonitrile degradation took place via the nitrilase pathway directly to benzoic acid without intermediate formation of benzamide. Samples with benzonitrilase activity of 7.6 nkat mg⁻¹ converted 3 mg benzonitrile in 1 h at 45°C. The half-life of benzonitrilase activity for a whole cell suspension and for cells immobilized in 2% agar was 4.5 min and 6 min at 70°C without substrate and 3 min at 90°C with substrate, respectively. The nitrilase had a broad substrate spectrum. The active biocatalyst obtained by immobilization was used in a continuous process and total biodegradation of 14.1 mM benzonitrile and 37.2 mM 4-cyanopyridine in a column bioreactor at 50°C for 5 h was achieved.