Post-transcriptional gene regulation: From genome-wide studies to principles

Abstract.: Post-transcriptional regulation of gene expression plays important roles in diverse cellular processes such as development, metabolism and cancer progression. Whereas many classical studies explored the mechanistics and physiological impact on specific mRNA substrates, the recent development of genome-wide analysis tools enables the study of post-transcriptional gene regulation on a global scale. Importantly, these studies revealed distinct programs of RNA regulation, suggesting a complex and versatile post-transcriptional regulatory network. This network is controlled by specific RNA-binding proteins and/or non-coding RNAs, which bind to specific sequence or structural elements in the RNAs and thereby regulate subsets of mRNAs that partly encode functionally related proteins. It will be a future challenge to link the spectra of targets for RNA-binding proteins to post-transcriptional regulatory programs and to reveal its physiological implication

Oxidative state in the estrus cycle of the buffaloes: a preliminary study

The oxidative status, the relationship between oxidizing agents (free radicals) and antioxidant system, is able to influence the correct performance of the estrus cycle. The aim of the experimentation was to define a first range of physiological values of reactive oxygen species (ROS) and the biological antioxidant potential (BAP), found during the phases of the buffalo estrus cycle. In this study, blood samples were taken from 30 buffaloes in various phases of the estrus cycle (estrus, diestrus, anestrus), on which serum determinations, by a photometer, of BAP and ROS have been carried out. The highest ROS values were detected during estrus and this is due to the primary role they play in determining ovulation. The highest levels of BAP have been found in diestrus, when they perform protective action against oxidative damage in the ovaries and uterus

Superficial angiomyxoma in a pregnant cow

A 3-year-old, pregnant, Alpine Brown cow showed a rapidly growing, pedunculated, skin mass located at the umbilical region, reaching 8 kg in weight over a 3-month period after its initial detection. Six days after parturition, the mass was completely surgically excised. During the follow-up period, the cow remained in good health, without signs of recurrence, and showed increased milk production. Histological examination of the mass revealed a loose proliferation of spindle-shaped or stellate cells, immersed in an abundant myxoid matrix with admixed numerous thin-walled blood vessels. Immunohistochemically, the tumour cells were positive for vimentin, α-smooth muscle actin, and desmin. Gross and histopathological features were compatible with superficial angiomyxoma, a subtype of angiomyxoma rarely described in humans, but not in the veterinary literature. The tumour did not infiltrate into the surrounding tissues, and there was no post-excision recurrence after 3 months. The possibility of hormonal dependence of the tumour during pregnancy is discussed based on such findings in some human cases

VP6-SUMO Self-Assembly as Nanocarriers for Gastrointestinal Delivery

High proteolytic degradation and poor absorption through epithelial barriers are major challenges to successful oral delivery of therapeutics. Nanoparticle platforms can enhance drug stability and extend the residence time in gastrointestinal (GI) tract. However, drug delivery systems are often inactivated in acidic environment of stomach or suffer poor absorption from intestinal cells due to the mucus layer. To overcome these issues we developed a drug delivery system constituted by a protein construct made by a Rotavirus capsid protein (VP6) and the small ubiquitin-like modifier SUMO. This chimeric construct allows specificity towards intestinal cells, the Rotavirus natural target, combined by an enhanced stability given by the eukaryotic protein transporter SUMO. Furthermore SUMO can act as a molecular switch that facilitates import/export of its ligand to the nucleus, the hypersensitive subcellular site target of many cell killing therapies. In this paper we show that SUMO-VP6 constructs self-assembly into stable nanocarriers. SUMO-VP6 nanocarriers display ideal features for drug delivery: a small size and high monodispersity, a high stability in different pH conditions and a high uptake in the nuclear and cytoplasmic compartment of intestinal cells. These features make SUMO-VP6 nanocarriers a promising novel system for oral delivery of poorly soluble drugs

Influence of indigenous starter coltures on the free fatty acids release during ripening in artisan sausages produced in the Basilicata region.

The influence of indigenous starter cultures on the free fatty acids content during ripening of \ubbsalsiccia\uab, a typical dry fermented sausage produced in the Basilicata region, was studied. Three batches of \ubbsalsiccia\uab were produced using different starter mixtures (Lactobacillus sakei G20 and Staphylococcus xylosus S81; L. sakei G20 and S. xylosus S142; L. sakei G20 and S. xylosus S206), while the control batch was produced without a starter. The amounts of free fatty acids present in the samples at the end of the ripening period were not significantly different, suggesting that the lipolytic enzymes naturally occurring in meat could play a predominant role in the free fatty acids release. Oleic and linoleic acids were present in the highest concentrations, while only small quantities of short chain fatty acids were detected, with acetic acid being the most representative one

Spin Splitting Tunable Optical Bandgap in GdN Thin Films for Spin Filtering

Rare-earth nitrides, such as gadolinium nitride (GdN), have great potential for spintronic devices due to their unique magnetic and electronic properties. GdN has a large magnetic moment, low coercitivity and strong spin polarization suitable for spin transistors, magnetic memories and spin-based quantum computing devices. Its large spin splitting of the optical bandgap functions as a spin-filter that offers the means for spin-polarized current injection into metals, superconductors, topological insulators, 2D layers and other novel materials. As spintronics devices require thin films, a successful implementation of GdN demands a detailed investigation of the optical and magnetic properties in very thin films. With this objective, we investigate the dependence of the direct and indirect optical bandgaps (Eg) of half-metallic GdN, using the trilayer structure AlN(10 nm)/GdN(t)/AlN(10 nm) for GdN film thickness t in the ranging from 6 nm to 350 nm, in both paramagnetic (PM) and ferromagnetic (FM) phases. Our results show a bandgap of 1.6 eV in the PM state, while in the FM state the bandgap splits for the majority (0.8 eV) and minority (1.2 eV) spin states. As the GdN film becomes thinner the spin split magnitude increases by 60%, going from 0.290 eV to 0.460 eV. Our results point to methods for engineering GdN films for spintronic devices

Benefits and risks of the hormetic effects of dietary isothiocyanates on cancer prevention

The isothiocyanate (ITC) sulforaphane (SFN) was shown at low levels (1-5 µM) to promote cell proliferation to 120-143% of the controls in a number of human cell lines, whilst at high levels (10-40 µM) it inhibited such cell proliferation. Similar dose responses were observed for cell migration, i.e. SFN at 2.5 µM increased cell migration in bladder cancer T24 cells to 128% whilst high levels inhibited cell migration. This hormetic action was also found in an angiogenesis assay where SFN at 2.5 µM promoted endothelial tube formation (118% of the control), whereas at 10-20 µM it caused significant inhibition. The precise mechanism by which SFN influences promotion of cell growth and migration is not known, but probably involves activation of autophagy since an autophagy inhibitor, 3-methyladenine, abolished the effect of SFN on cell migration. Moreover, low doses of SFN offered a protective effect against free-radical mediated cell death, an effect that was enhanced by co-treatment with selenium. These results suggest that SFN may either prevent or promote tumour cell growth depending on the dose and the nature of the target cells. In normal cells, the promotion of cell growth may be of benefit, but in transformed or cancer cells it may be an undesirable risk factor. In summary, ITCs have a biphasic effect on cell growth and migration. The benefits and risks of ITCs are not only determined by the doses, but are affected by interactions with Se and the measured endpoint

Upregulation of the cell-cycle regulator RGC-32 in Epstein-Barr virus-immortalized cells

Epstein-Barr virus (EBV) is implicated in the pathogenesis of multiple human tumours of lymphoid and epithelial origin. The virus infects and immortalizes B cells establishing a persistent latent infection characterized by varying patterns of EBV latent gene expression (latency 0, I, II and III). The CDK1 activator, Response Gene to Complement-32 (RGC-32, C13ORF15), is overexpressed in colon, breast and ovarian cancer tissues and we have detected selective high-level RGC-32 protein expression in EBV-immortalized latency III cells. Significantly, we show that overexpression of RGC-32 in B cells is sufficient to disrupt G2 cell-cycle arrest consistent with activation of CDK1, implicating RGC-32 in the EBV transformation process. Surprisingly, RGC-32 mRNA is expressed at high levels in latency I Burkitt's lymphoma (BL) cells and in some EBV-negative BL cell-lines, although RGC-32 protein expression is not detectable. We show that RGC-32 mRNA expression is elevated in latency I cells due to transcriptional activation by high levels of the differentially expressed RUNX1c transcription factor. We found that proteosomal degradation or blocked cytoplasmic export of the RGC-32 message were not responsible for the lack of RGC-32 protein expression in latency I cells. Significantly, analysis of the ribosomal association of the RGC-32 mRNA in latency I and latency III cells revealed that RGC-32 transcripts were associated with multiple ribosomes in both cell-types implicating post-initiation translational repression mechanisms in the block to RGC-32 protein production in latency I cells. In summary, our results are the first to demonstrate RGC-32 protein upregulation in cells transformed by a human tumour virus and to identify post-initiation translational mechanisms as an expression control point for this key cell-cycle regulator