RNA silencing platforms in plants

Since the discovery of RNAi, its mechanism in plants and animals has been intensively studied, widely exploited as a research tool, and used for a number of potential commercial applications. In this article, we discuss the platforms for delivering RNAi in plants. We provide a brief background to these platforms and concentrate on discussing the more recent advances, comparing the RNAi technologies used in plants with those used in animals, and trying to predict the ways in which RNAi technologies may further develop.     

Virulence Factors and Antifungal Susceptibility in Candida Species Isolated from Dermatomycosis Patients

Mycopathologia - Dermatomycoses caused by Candida spp. are increasingly common, however there are few reports in the literature regarding their epidemiology, pathogenesis and antifungal...     

Scanning X‐ray microdiffraction of decellularized pericardium tissue at increasing glucose concentration

Blood glucose supplies energy to cells and is critical for the human brain. Glycation of collagen, the nonenzymatic formation of glucose‐bridges, relates to diseases of aging populations and diabetics. This chemical reaction, together with its biomechanical effects, has been well studied employing animal models. However, the direct impact of glycation on collagen nano‐structure is largely overlooked, and there is a lack of ex vivo model systems. Here, we present the impact of glucose on collagen nanostructure in a model system based on abundantly available connective tissue of farm animals. By combining ex vivo small and wide‐angle X‐ray scattering (SAXS/WAXS) imaging, we characterize intra‐ and inter‐molecular parameters of collagen in decellularized bovine pericardium with picometer precision. We observe three distinct regimes according to glucose concentration. Such a study opens new avenues for inspecting the effects of diabetes mellitus on connective tissues and the influence of therapies on the resulting secondary disorders.      

Effect of Simultaneous Administration of Dihydroxyacetone on the Diffusion of Lawsone Through Various In Vitro Skin Models

Unprotected sunlight exposure is a risk factor for a variety of cutaneous cancers. Topically used dihydroxyacetone (DHA) creates, via Maillard reaction, chemically fixed keratin sunscreen in the stratum corneum with significant protection against UVA/Soret radiation. When used in conjunction with naphthoquinones a naphthoquinone-modified DHA Maillard reaction is produced that provides protection across the UVB/UVA/Soret spectra lasting up to 1 week, resisting sweating and contact removal. The aim of this study was to examine a simplified version of this formulation for effect on UV transmission and to determine if penetration levels merit toxicity concerns. Permeability was demonstrated for freshly prepared DHA (30 mg/mL) and lawsone (0.035 mg/mL) alone and in combination using a side-by-side diffusion apparatus at 37°C over 48 h across shed snake skin and dermatomed pig skin. These samples were then examined for effectiveness and safety. Concentrations were determined by HPLC and UPLC monitored from 250–500 nm. Lawsone flux significantly decreased across pig skin (20.8 (±4.8) and 0.09 (±0.1) mg/cm² h without and with DHA, respectively) but did not change across shed snake skin in the presence of DHA. Significantly reduced lawsone concentration was noted in donor chambers of combined solutions. Damage was not observed in any skins. Darker coloration with greater UV absorbance was observed in skins exposed to the combined solution versus individual solutions. This study confirmed that combined DHA and lawsone provided effective blocking of ultraviolet light through products bound in keratinized tissue. DHA permeation levels in pig skin suggest further in vitro and in vivo study is required to determine the safety of this system.KEY WORDS: cancer, dihydroxyacetone, FAMMM, naphthoquinone, skin     

Phylogeography and evolutionary history of reassortant H9N2 viruses with potential human health implications

Avian influenza viruses of the H9N2 subtype have seriously affected the poultry industry of the Far and Middle East since the mid-1990s and are considered one of the most likely candidates to cause a new influenza pandemic in humans. To understand the genesis and epidemiology of these viruses, we investigated the spatial and evolutionary dynamics of complete genome sequences of H9N2 viruses circulating in nine Middle Eastern and Central Asian countries from 1998 to 2010. We identified four distinct and cocirculating groups (A, B, C, and D), each of which has undergone widespread inter- and intrasubtype reassortments, leading to the generation of viruses with unknown biological properties. Our analysis also suggested that eastern Asia served as the major source for H9N2 gene segments in the Middle East and Central Asia and that in this geographic region within-country evolution played a more important role in shaping viral genetic diversity than migration between countries. The genetic variability identified among the H9N2 viruses was associated with specific amino acid substitutions that are believed to result in increased transmissibility in mammals, as well as resistance to antiviral drugs. Our study highlights the need to constantly monitor the evolution of H9N2 viruses in poultry to better understand the potential risk to human health posed by these viruses.     

The evolution and diversification of Dicers in plants

Most multicellular organisms regulate developmental transitions by microRNAs, which are generated by an enzyme, Dicer. Insects and fungi have two Dicer-like genes, and many animals have only one, yet the plant, Arabidopsis, has four. Examining the poplar and rice genomes revealed that they contain five and six Dicer-like genes, respectively. Analysis of these genes suggests that plants require a basic set of four Dicer types which were present before the divergence of mono- and dicotyledonous plants ( approximately 200 million years ago), but after the divergence of plants from green algae. A fifth type of Dicer seems to have evolved in monocots.     

Natural and commercial Salix clones differ in their ecophysiological response to Zn stress

This study was carried out to determine the effect of different zinc concentrations on the ecophysiological response of Salix clones: four commercial clones (“1962”, “1968”, “Drago”, and “Levante”) selected for short rotation coppice, and one natural clone, “Sacco”, obtained from a contaminated area. Gas exchanges, chlorophyll a fluorescence (JIP-test), relative chlorophyll content, and biometric parameters were measured in plants grown for fifteen days in soil containing Zn concentrations of 0, 300, 750, and 1,500 mg(ZnCl₂) kg^?1. Ecophysiological response to metal stress differed in dependence on the Zn concentration and clone. At the low Zn concentration (300 mg kg^?1), the absence of any significant reductions in parameters investigated indicated an efficient plant homeostasis to maintain the metal content within phytotoxic limits. Stomatal limitation, observed at 750 and 1,500 mg kg^?1, which was found in all clones after three days of the treatment, might be caused by indirect effects of metal on guard cells. Among commercial clones, “Drago” was more sensitive to Zn stress, showing inhibition of growth, while “1962” clone showed a downregulation of PSII photochemistry following the slowdown in the Calvin-Benson cycle. On the contrary, the natural Salix clone (“Sacco”) performed better compared to the other clones due to activation of a photosynthetic compensatory mechanism.