Converging Strategies in Expression of Human Complex Retroviruses

The discovery of human retroviruses in the early 1980s revealed the existence of viral-encoded non-structural genes that were not evident in previously described animal retroviruses. Based on the absence or presence of these additional genes retroviruses were classified as ‘simple’ and ‘complex’, respectively. Expression of most of these extra genes is achieved through the generation of alternatively spliced mRNAs. The present review summarizes the genetic organization and expression strategies of human complex retroviruses and highlights the converging mechanisms controlling their life cycles

Post-transcriptional regulation of HTLV gene expression: Rex to the rescue

Human T-lymphotropic virus type 1 (HTLV-1) and other members of the Deltaretrovirus genus code for a regulatory protein named Rex that binds to the Rex-responsive element present on viral mRNAs. Rex rescues viral mRNAs from complete splicing or degradation and guides them to the cytoplasm for translation. The activity of Rex is essential for expression of viral transcripts coding for the virion components and thus represents a potential target for virus eradication. We present an overview of the functional properties of the HTLV-1 and HTLV-2 Rex proteins (Rex-1 and Rex-2), outline mechanisms controlling Rex function, and discuss similarities and differences in the sequences of Rex coded by HTLV-1, -2, -3, and -4 that may influence their molecular anatomy and functional properties

An introduction to the early Holocene eolian deposits of Grotta Romanelli, Apulia, Southern Italy

Due to its geographic position and geomorphological configuration, Grotta Romanelli acted as a sediment trap since at least MIS 5. The so-called 'terre brune' sequence is a deposit mainly of eolian origin bearing upper Palaeolithic artefacts and fossil re-mains of vertebrate fauna; it was deposited during the Glacial-Interglacial transition and the Holocene. Sedimentology and mineralogy of this deposit are investigated. The stratigraphic sequence provides a promising archive within which both human and climatic impacts can be studied

Far-Field Pattern Reconstruction from Near-Field Data Collected via a Nonconventional Plane-Rectangular Scanning: Experimental Testing

This paper deals with the experimental validation of an efficient near-field-far-field (NF-FF) transformation using the planar wide-mesh scanning (PWMS). Such a nonconventional plane-rectangular scanning technique is so named, since the sample grid is characterized by meshes wider and wider when going away from the center, and makes it possible to lower the number of needed measurements, as well as the time required for the data acquisition when dealing with quasi-planar antennas. It relies on the use of the nonredundant sampling representations of electromagnetic fields which employ an oblate ellipsoid or a surface formed by two circular "bowls" with the same aperture diameter but eventually different bending radii to shape a quasi-planar antenna. A two-dimensional optimal sampling interpolation formula allows the reconstruction of the NF data at any point on the measurement plane and, in particular, at those required by the classical NF-FF transformation with the conventional plane-rectangular scanning. The measurements, performed at the planar NF facility of the antenna characterization laboratories of Selex ES, have confirmed the effectiveness of this innovative scanning also from the experimental viewpoint

Expression of alternatively spliced human T-cell leukemia virus type 1 mRNAs is influenced by mitosis and by a novel cis-acting regulatory sequence

Human T-cell leukemia virus type 1 (HTLV-1) expression depends on the concerted action of Tax, which drives transcription of the viral genome, and Rex, which favors expression of incompletely spliced mRNAs and determines a 2-phase temporal pattern of viral expression. In the present study, we investigated the Rex dependence of the complete set of alternatively spliced HTLV-1 mRNAs. Analyses of cells transfected with Rex-wild-type and Rex-knockout HTLV-1 molecular clones using splice site-specific quantitative reverse transcription (qRT)-PCR revealed that mRNAs encoding the p30Tof, p13, and p12/8 proteins were Rex dependent, while the p21rex mRNA was Rex independent. These findings provide a rational explanation for the intermediate-late temporal pattern of expression of the p30tof, p13, and p12/8 mRNAs described in previous studies. All the Rex-dependent mRNAs contained a 75-nucleotide intronic region that increased the nuclear retention and degradation of a reporter mRNA in the absence of other viral sequences. Selective 2'-hydroxyl acylation analyzed by primer extension (SHAPE) analysis revealed that this sequence formed a stable hairpin structure. Cell cycle synchronization experiments indicated that mitosis partially bypasses the requirement for Rex to export Rex-dependent HTLV-1 transcripts. These findings indicate a link between the cycling properties of the host cell and the temporal pattern of viral expression/latency that might influence the ability of the virus to spread and evade the immune system

Dyes of a Shadow Theatre: Investigating Tholu Bommalu Indian Puppets through a Highly Sensitive Multi-Spectroscopic Approach

Tholu Bommalu are typical leather puppets of the traditional Indian shadow theatre. Two of these objects are part of a collection in the International Puppets Museum “Antonio Pasqualino” (Palermo, Sicily, Italy), which can count on one hundred-seventy-three of artifacts. These Indian puppets were investigated to obtain information related to the use of dyes for their manufacturing through a multi-technical approach exploiting the combination of highly sensitive spectroscopic techniques. Wet cotton stubbons were used to entrap small particles of dyes on the fibers from the art objects for the consequent analyses. Visible Light Micro-Reflectance spectroscopy was employed for the preliminary identification of the molecular class of dyes directly on the swabs, while Surface Enhanced Raman Scattering allowed the identification of the specific dye. Several synthetic dyes belonging to different typologies of coloring compounds were identified. The study resulted in an interesting overview of dyes used in recent Tholu Bommalata manufacturing through the combination of micro-invasive techniques directly on the sampling substrate

Biochemical responses, feeding and survival in the solitary bee Osmia bicornis following exposure to an insecticide and a fungicide alone and in combination

In agricultural ecosystems, bees are exposed to combinations of pesticides that may have been applied at different times. For example, bees visiting a flowering crop may be chronically exposed to low concentrations of systemic insecticides applied before bloom and then to a pulse of fungicide, considered safe for bees, applied during bloom. In this study, we simulate this scenario under laboratory conditions with females of the solitary bee, Osmia bicornis L. We studied the effects of chronic exposure to the neonicotinoid insecticide, Confidor (R) (imidacloprid) at a realistic concentration, and of a pulse (1 day) exposure of the fungicide Folicur (R) SE (tebuconazole) at field application rate. Syrup consumption, survival, and four biomarkers: acetylcholinesterase (AChE), carboxylesterase (CaE), glutathione S-transferase (GST), and alkaline phosphatase (ALP) were evaluated at two different time points. An integrated biological response (IBRv2) index was elaborated with the biomarker results. The fungicide pulse had no impact on survival but temporarily reduced syrup consumption and increased the IBRv2 index, indicating potential molecular alterations. The neonicotinoid significantly reduced syrup consumption, survival, and the neurological activity of the enzymes. The co-exposure neonicotinoid-fungicide did not increase toxicity at the tested concentrations. AChE proved to be an efficient biomarker for the detection of early effects for both the insecticide and the fungicide. Our results highlight the importance of assessing individual and sub-individual endpoints to better understand pesticide effects on bees

Biochemical responses, feeding and survival in the solitary bee Osmia bicornis following exposure to an insecticide and a fungicide alone and in combination

In agricultural ecosystems, bees are exposed to combinations of pesticides that may have been applied at different times. For example, bees visiting a flowering crop may be chronically exposed to low concentrations of systemic insecticides applied before bloom and then to a pulse of fungicide, considered safe for bees, applied during bloom. In this study, we simulate this scenario under laboratory conditions with females of the solitary bee, Osmia bicornis L. We studied the effects of chronic exposure to the neonicotinoid insecticide, Confidor® (imidacloprid) at a realistic concentration, and of a pulse (1 day) exposure of the fungicide Folicur® SE (tebuconazole) at field application rate. Syrup consumption, survival, and four biomarkers: acetylcholinesterase (AChE), carboxylesterase (CaE), glutathione S-transferase (GST), and alkaline phosphatase (ALP) were evaluated at two different time points. An integrated biological response (IBRv2) index was elaborated with the biomarker results. The fungicide pulse had no impact on survival but temporarily reduced syrup consumption and increased the IBRv2 index, indicating potential molecular alterations. The neonicotinoid significantly reduced syrup consumption, survival, and the neurological activity of the enzymes. The co-exposure neonicotinoid-fungicide did not increase toxicity at the tested concentrations. AChE proved to be an efficient biomarker for the detection of early effects for both the insecticide and the fungicide. Our results highlight the importance of assessing individual and sub-individual endpoints to better understand pesticide effects on bees

The GSK3\u3b2 inhibitor BIS I reverts YAP-dependent EMT signature in PDAC cell lines by decreasing SMADs expression level

The Yes-associated protein, YAP, is a transcriptional co-activator, mediating the Epithelial to Mesenchymal Transition program in pancreatic ductal adenocarcinoma (PDAC). With the aim to identify compounds that can specifically modulate YAP functionality in PDAC cell lines, we performed a small scale, drug-based screening experiment using YAP cell localization as the read-out. We identified erlotinib as an inducer of YAP cytoplasmic localization, an inhibitor of the TEA luciferase reporter system and the expression of the bona fide YAP target gene, Connective Tissue Growth Factor CTGF. On the other hand, BIS I, an inhibitor of PKC\u3b4 and GSK3\u3b2, caused YAP accumulation into the nucleus. Activation of \u3b2-catenin reporter and interfering experiments show that inhibition of the PKC\u3b4/GSK3\u3b2 pathway triggers YAP nuclear accumulation inducing YAP/TEAD transcriptional response. Inhibition of GSK3\u3b2 by BIS I reduced the expression levels of SMADs protein and reduced YAP contribution to EMT. Notably, BIS I reduced proliferation, migration and clonogenicity of PDAC cells in vitro, phenocopying YAP genetic down-regulation. As shown by chromatin immunoprecipitation experiments and YAP over-expressing rescue experiments, BIS I reverted YAP-dependent EMT program by modulating the expression of the YAP target genes E-cadherin, vimentin, CTGF and of the newly identified target, CD133. In conclusion, we identified two different molecules, erlotinib and BIS I, modulating YAP functionality although via different mechanisms of action, with the second one specifically inhibiting the YAP-dependent EMT program in PDAC cell lines