Reference genes for RT-qPCR normalisation in different tissues, developmental stages and stress conditions of amaranth.

Studies of gene expression are very important for the identification of genes that participate in different biological processes. Currently, reverse transcription quantitative real-time PCR (RT-qPCR) is a high-throughput, sensitive and widely used method for gene expression analysis. Nevertheless, RT-qPCR requires precise normalisation of data to avoid the misinterpretation of experimental data. In this sense, the selection of reference genes is critical for gene expression analysis. At this time, several studies focus on the selection of reference genes in several species. However, the identification and validation of reference genes for the normalisation of RT-qPCR have not been described in amaranth. A set of seven housekeeping genes were analysed using RT-qPCR, to determine the most stable reference genes in amaranth for normalisation of gene expression analysis. Transcript stability and gene expression level of candidate reference genes were analysed in different tissues, at different developmental stages and under different types of stress. The data were compared using the geNorm, NormFinder and Bestkeeper statistical methods. The reference genes optimum for normalisation of data varied with respect to treatment. The results indicate that AhyMDH, AhyGAPDH, AhyEF-1α and AhyACT would be optimum for accurate normalisation of experimental data, when all treatment are analysed in the same experiment. This study presents the most stable reference genes for normalisation of gene expression analysis in amaranth, which will contribute significantly to future gene studies of this species.

Severe malaria in immigrant haematological patient.

Severe malaria is a life-threatening condition caused by Plasmodium falciparum. Rupture of red blood cells when merozoites release to the bloodstream is responsible for the clinical manifestations, febrile fever reaching 39 °C, and other unspecific symptoms. P. falciparum is considered as the worst form of malaria. Moreover, this species has cytoadherence to red blood cells. This can lead to an organic dysfunction. People coming from hyper endemic areas have developed a partial immunity, but immunodepressed people are a group with a greater risk. Due to the high mortality rate associated to this disease, early diagnosis and a prompt treatment implementation are essential. However, the missed or delayed diagnosis is one of the major reasons of reaching a severe malaria disease. This case reflects the complexity of the diagnosis in an immigrant and immunodepressed patient with a haematological neoplasm with a severe infection by P. falciparum due to the unspecified symptoms and the overlapping of the same.

A comparison of analytical techniques for evaluating food waste degradation by anaerobic digestion.

Organic matter contained in food waste was degraded by anaerobic digestion under mesophilic and thermophilic conditions at two hydraulic retention times. Evolution of the digestion process was followed by thermogravimetry analysis, fluorescence spectroscopy and (1)H nuclear magnetic resonance. All analytical methods suggested that longer retention times might be required for food waste stabilization under mesophilic conditions as compared to thermophilic stabilization. All the analytical methods showed that the stabilization process consisted of two steps, where complex organic molecules were formed during initial stabilization and then digested providing sufficient hydraulic retention time. Longer hydraulic retention times were required for food waste stabilization under mesophilic conditions. Overall, thermal and (1)H NMR analyses of the digestate samples might be recommended if more detailed analysis is required, while fluorescence measurements can be used as a fast screening technique, which provides qualitative assessment of the stabilization process.