MicroRNA from moringa oleifera : identification by high throughput sequencing and their potential contribution to plant medicinal value

Moringa oleifera is a widespread plant with substantial nutritional and medicinal value. We postulated that microRNAs (miRNAs), which are endogenous, noncoding small RNAs regulating gene expression at the post-Transcriptional level, might contribute to the medicinal properties of plants of this species after ingestion into human body, regulating human gene expression. However, the knowledge is scarce about miRNA in Moringa. Furthermore, in order to test the hypothesis on the pharmacological potential properties of miRNA, we conducted a high-Throughput sequencing analysis using the Illumina platform. A total of 31,290,964 raw reads were produced from a library of small RNA isolated from M. oleifera seeds. We identified 94 conserved and two novel miRNAs that were validated by qRT-PCR assays. Results from qRT-PCR trials conducted on the expression of 20 Moringa miRNA showed that are conserved across multiple plant species as determined by their detection in tissue of other common crop plants. In silico analyses predicted target genes for the conserved miRNA that in turn allowed to relate the miRNAs to the regulation of physiological processes. Some of the predicted plant miRNAs have functional homology to their mammalian counterparts and regulated human genes when they were transfected into cell lines. To our knowledge, this is the first report of discovering M. oleifera miRNAs based on highthroughput sequencing and bioinformatics analysis and we provided new insight into a potential cross-species control of human gene expression. The widespread cultivation and consumption of M. oleifera, for nutritional and medicinal purposes, brings humans into close contact with products and extracts of this plant species. The potential for miRNA transfer should be evaluated as one possible mechanism of action to account for beneficial properties of this valuable species

Bilateral Investment Treaties and the European Union. Recent Developments in Arbitration and Before the ECJ

The issue of the relationship between Bilateral Investment Treaties (BITs) and the EU legal order has recently attracted attention amongst scholars and practitioners in the field of international investment arbitration. Under a first perspective of the problem, the Arbitral Tribunal in Eastern Sugar B.V. v. The Czech Republic was confronted with the question of whether there was any room left for BITs between EU Member States. The Tribunal discussed the legal arguments advanced for and against the applicability of such \u201cintra-EU BITs\u201d between Member States. The issue, which is particularly relevant considering that there are currently more than 190 BITs concluded between EU Member States, will be analysed in the first part of this article. Under a second point of view of the problem, the European Court of Justice (ECJ) handed down two judgments on 3 March 2009 addressing incompatibilities with EC Law resulting from certain BITs entered into by Sweden and Austria with third countries. The second part of this article will deal with the consequences arising out of the Court\u2019s rulings with regard to existing and future BITs entered into by Member States with third countries

Internalization and intracellular retention of CD4 are two separate functions of the human immunodeficiency virus type 1 Nef protein

The pathogenic Nef protein of the human immunodeficiency virus type 1 (HIV-1) downregulates CD4 by inducing its endocytosis and by inhibiting the transport of the receptor to the cell membrane. By means of in vivo-selected mutations, we show that L37, P78 and El 77 residues of Nef are required for its effect on CD4 internalization and recycling but dispensable for Nef-induced retention and degradation of intracellular CD4. Of note, the function of Nef on the anterograde transport of newly synthesized CD4 molecules is irrelevant in cells with a slow constitutive CD4 turnover such as T cell lines. Moreover, we show that a mutated CD4 that is unresponsive to Nef-mediated endocytosis, CD4LL(144)AA, is retained intracellularly and degraded by Nef like wild-type CD4. Thus, Nef's abilities to enhance endocytosis and induce intracellular retention of CD4 are mediated by separate protein surfaces and occur through distinct mechanisms