Long‐duration time series of the chemical composition of fumaroles and of soil CO2 flux reveal that important variations in the activity of the Solfatara fumarolic field, the most important ...hydrothermal site of Campi Flegrei, occurred in the 2000–2008 period. A continuous increase of the CO2 concentrations and a general decrease of the CH4 concentrations are interpreted to be the consequence of the increment of the relative amount of magmatic fluids, rich in CO2 and poor in CH4, hosted by the hydrothermal system. Contemporaneously, the H2O‐CO2‐He‐N2 gas system shows remarkable compositional variations in the samples collected after July 2000 with respect to the previous ones, indicating the progressive arrival at the surface of a magmatic component different from that involved in the 1983–1984 episode of volcanic unrest (1983–1984 bradyseism). The change starts in 2000, concurrently with the occurrence of relatively deep, long‐period seismic events which were the indicator of the opening of an easy ascent pathway for the transfer of magmatic fluids toward the shallower, brittle domain hosting the hydrothermal system. Since 2000, this magmatic gas source is active and causes ground deformations and seismicity as well as the expansion of the area affected by soil degassing of deeply derived CO2. Even though the activity will most probably be limited to the expulsion of large amounts of gases and thermal energy, as observed in other volcanoes and in the past activity of Campi Flegrei, the behavior of the system in the future is, at the moment, unpredictable.
Systematic and timely documentation of triggered (i.e. event) landslides is fundamental to build extensive datasets worldwide that may help define and/or validate trends in response to climate ...change. More in general, preparation of landslide inventories is a crucial activity since it provides the basic data for any subsequent analysis. In this work we present an event landslide inventory map (E-LIM) that was prepared through a systematic reconnaissance field survey in about 1 month after an extreme rainfall event hit an area of about 5000 km
in the Marche-Umbria regions (central Italy). The inventory reports evidence of 1687 triggered landslides in an area of ~550 km
. All slope failures were classified according to type of movement and involved material, and documented with field pictures, wherever possible. The database of the inventory described in this paper as well as the collection of selected field pictures associated with each feature is publicly available at figshare.
•We present the first measurements of soil C6H6 fluxes in a volcanic system.•Methane oxidation rate is controlled by soil gas fluxes.•Benzene oxidation rate is controlled by presence of a SO42--rich ...aquifer.•Fumarolic emissions cause a strong benzene air contamination at a local scale.•Endogenous monoaromatics are detected in air samples from the whole crater.
Measurements of soil fluxes of hydrothermal gases, with special emphasis on C6H6, as well as chemical composition of mono-aromatic compounds in fumaroles and air, were carried out in April 2012 at the Solfatara crater (Campi Flegrei, Southern Italy) to investigate the distribution and behavior of these species as they migrate through the soil from their deep source to the atmosphere. Soil fluxes of CO2, CH4 and C6H6 exhibit good spatial correlation, suggesting that diffuse degassing is mainly controlled by local fractures. The calculated total output of diffuse C6H6 from Solfatara is 0.10kgday−1, whereas fluxes of CO2 and CH4 are 79×103 and 1.04kgday−1, respectively. A comparison between soil gas fluxes and fumarole composition reveals that within the crater soil CH4 is significantly affected by oxidation processes, which are more efficient for low gas fluxes, being dependent on the residence time of the uprising hydrothermal gases at shallow depth. Benzene degradation, mainly proceeding through oxidation via benzoate, seems to be strongly controlled by the presence of a shallow SO42--rich aquifer located in the central and southwestern sectors of the crater, suggesting that the process is particularly efficient when SO42- acts as terminal electron acceptor (SO4 reduction). Relatively high C6H6/C7H8 ratios, typical of hydrothermal fluids, were measured in air close to the main fumarolic field of Solfatara crater. Here, C6H6 concentrations, whose detection limit is ∼0.1μgm−3, are more than one order of magnitude higher than the limit value for ambient air (5μgm−3). This suggests that hydrothermal fluids have a strong impact on air quality in the immediate surroundings of the fumarolic vents. Significant concentrations of endogenous mono-aromatics were also detected in air samples collected from the northern and western sides of the crater, where these gas compounds are mostly fed by diffuse degassing through the crater bottom soil.
Antisense oligonucleotide drug design Schiavone, N; Donnini, M; Nicolin, A ...
Current pharmaceutical design,
03/2004, Letnik:
10, Številka:
7
Journal Article
Recenzirano
Maneuvering single gene expression is not only an optimal way to study gene function but also an ambitious goal, which will lead to the treatment of a variety of human diseases whose main ...pathogenetic event is a genetic alteration. The recent efforts focusing on the genome project have led to array based, high throughput, gene expression analysis techniques that allow the study of complex molecular networks. Combining these powerful new technologies with modulation of gene expressions is making it possible to unravel complex molecular networks or, vice versa, to find new gene products responsible for pathological conditions on which exogenous modulation can be productive. Efficient and specific modulation of gene expression can be obtained either by producing transgenic or gene knockout organisms or cells (gene targeting), or by treating organisms or cells with short synthetic nucleic acid segments in antisense orientation with respect to the targeted mRNAs (mRNA targeting by antisense strategy). While genome manipulation is a time consuming and expensive approach, requiring invasive intervention, the "antisense strategy" is characterized by high flexibility resulting from safeness, specificity, reversibility, modulability, and low cost. The rationale of the antisense strategy is that, once one gene sequence is known, its expression can be silenced by application of synthetic single-strand nucleic acid segments (oligonucleotides) whose sequence is in antisense orientation compared to the targeted mRNA. Recently, this "informational" strategy has been boosted by the discovery of the RNA interference: a natural mechanism by which cells are thought to fight detrimental exogenous viruses and endogenous transposons. Despite promising futures, antisense-based therapeutics are far from being an established reality. This review analyses the recent improvements in antisense-based gene expression modulation, focuses on the treatment of diseases in the light of the past, and provides our personal findings on this topic.
The permeability transition pore (PTP) is a mitochondrial channel whose opening causes the mitochondrial membrane potential (ΔΨ) collapse that leads to apoptosis. Some ubiquinone analogues have been ...demonstrated previously to modulate the PTP open-closed transition in isolated mitochondria and thought to act through a common PTP-binding site rather than through oxidation-reduction reactions. We have demonstrated recently both in vitro and in vivo that the ubiquitous free radical scavenger and respiratory chain coenzyme Q10 (CoQ10) prevents keratocyte apoptosis induced by excimer laser irradiation more efficiently than other antioxidants. On this basis, we hypothesized that the antiapoptotic property of CoQ10 could be independent of its free radical scavenging ability and related to direct inhibition of PTP opening. In this study, we have verified this hypothesis by evaluating the antiapoptotic effects of CoQ10 in response to apoptotic stimuli, serum starvation, antimycin A, and ceramide, which do not generate free radicals, in comparison to control, free radical-generating UVC irradiation. As hypothesized, CoQ10 dramatically reduced apoptotic cell death, attenuated ATP decrease, and hindered DNA fragmentation elicited by all apoptotic stimuli. This was accompanied by inhibition of mitochondrial depolarization, cytochrome c release, and caspase 9 activation. Because these events are consequent to mitochondrial PTP opening, we suggest that the antiapoptotic activity of CoQ10 could be related to its ability to prevent this phenomenon.
Defects in apoptosis (programmed cell death) have recently emerged as being closely involved in the pathogenesis of most ocular diseases and, therefore, apoptosis is now a topic of exponential ...interest in ophthalmology. This review summarizes recent works on mechanisms of apoptosis, from its initiation and modulation to the switching-on of its execution machinery. Interactions of cell death with cell division programs to orchestrate ontogenesis, aging, and adult life and their alterations in human diseases are pointed out. Two main apoptotic signaling pathways are identified: a death receptor-dependent (extrinsic) pathway and a mitochondrion-dependent (intrinsic) pathway. Mitochondrion harbors both antiapoptotic (Bcl-2, Bcl-XL) and apoptotic factors (Smac/Diablo, Apaf-1, cytochrome c). Its permeability transition pore (mPTP) is the main trigger of cell suicide. The process of mPTP opening, in association with extrusion to cytoplasm of a variety of apoptotic factors, is shown. Cytochrome c is one of these apoptotic factors. When expelled to cytoplasm, this double-faced respiratory chain component assembles with two other modules, Apaf-1 and procaspase 9, to form a protein complex--the apoptosome--that starts apoptosis execution. Another respiratory chain component, the CoQ10, is believed to counteract mPTP opening. What makes apoptosis particularly exciting for medicine is that its dysfunctions play a central role in the pathogenesis of several human diseases. For instance, excesses of apoptosis lead to cell loss that accompanies neurodegenerative diseases, whereas genetically determined defects of apoptosis lead to the deregulated cell proliferation typical of cancer. A variety of ophthalmologic diseases, such as post-keratectomy haze, corneal lesions, cataract, glaucoma, senile maculopathies, and genetic ocular pathologies, that underlie apoptosis dysfunctions are treated in detail in the other reviews of this issue.
We previously identified a conserved A + U-rich element (ARE) in the 3′-untranslated region of bcl-2mRNA. We have also recently demonstrated that the bcl-2ARE interacts with a number of ARE-binding ...proteins (AUBPs) whose pattern changes during apoptosis in association with bcl-2mRNA half-life reduction. Here we show that the AUBP AUF1 bindsin vitro to bcl-2 mRNA. The results obtained in a yeast RNA three-hybrid system have demonstrated that the 1–257-amino acid portion of p37 AUF1 (conserved in all isoforms), containing the two RNA recognition motifs, also binds to thebcl-2 ARE in vivo. UVC irradiation-induced apoptosis results in an increase of AUF1. Inhibition of apoptosis by a general caspase inhibitor reduces this increase by 2–3-fold. These results indicate involvement of AUF1 in the ARE/AUBP-mediated modulation of bcl-2 mRNA decay during apoptosis.
To assess in vitro the potential of the free radical scavenger ubiquinone Q10 in preventing keratocyte apoptosis after argon fluoride (ArF) excimer laser irradiation.
Cultured rabbit keratocytes were ...irradiated at very low single-pulse laser fluences. The cumulative effects generated by three total fluence doses between 12 and 45 mJ/cm2, representative of single-pulse subablative doses during photorefractive keratectomy (PRK) in humans, were evaluated. We employed the following parameters to compare pretreated (10 microM ubiquinone Q10) and untreated samples: 1) number and morphology of living cells by Trypan blue test and ultramicroscopy, respectively; 2) level of free-radical formation assessed by malonaldehyde quantitation; 3) cellular energy level evaluated by ATP assay.
Excimer laser irradiation kills cultured keratocytes by inducing apoptosis. The effect increases with the cumulative fluence dose. In the samples pretreated with ubiquinone Q10 there were significantly fewer cumulative apoptotic events than in the untreated ones. Quantitative analysis of malonaldehyde cellular levels suggested this protective action of ubiquinone Q10 was connected with its ability to scavenge laser-generated free radicals. ATP assay also confirmed that it raised cellular energy levels.
The treatment of corneal keratocytes with relatively low concentrations of ubiquinone Q10 can prevent apoptosis after ArF excimer laser irradiation. If these findings are confirmed on human keratocytes this treatment could be usefully exploited in the PRK surgical procedure. That might lead to a reduction in the occurrence of haze and curvature regression triggered by programmed cell death.
Modulation of mRNA stability by regulatory cis-acting AU-rich elements (AREs) and ARE-binding proteins is an important posttranscriptional mechanism of gene expression control. We previously ...demonstrated that the 3'-untranslated region of BCL-2 mRNA contains an ARE that accounts for rapid BCL-2 down-regulation in response to apoptotic stimuli. We also demonstrated that the BCL-2 ARE core interacts with a number of ARE-binding proteins, one of which is AU-rich factor 1/heterogeneous nuclear ribonucleoprotein D, known for its interaction with mRNA elements of others genes. In an attempt to search for other BCL-2 mRNA-binding proteins, we used the yeast RNA three-hybrid system assay and identified a novel human protein that interacts with BCL-2 ARE. We refer to it as TINO. The predicted protein sequence of TINO reveals two amino-terminal heterogeneous nuclear ribonucleoprotein K homology motifs for nucleic acid binding and a carboxyl-terminal RING domain, endowed with a putative E3 ubiquitin-protein ligase activity. In addition the novel protein is evolutionarily conserved; the two following orthologous proteins have been identified with protein-protein BLAST: posterior end mark-3 (PEM-3) of Ciona savignyi and muscle excess protein-3 (MEX-3) of Caenorhabditis elegans. Upon binding, TINO destabilizes a chimeric reporter construct containing the BCL-2 ARE sequence, revealing a negative regulatory action on BCL-2 gene expression at the posttranscriptional level.
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