Targeting mGlu5 metabotropic glutamate receptors in the treatment of cognitive dysfunction in a mouse model of phenylketonuria

We studied group-I metabotropic glutamate (mGlu) receptors in Pah(enu2) (ENU2) mice, which mimic the genetics and neurobiology of human phenylketonuria (PKU), a metabolic disorder characterized, if untreated, by autism, and intellectual disability (ID). Male ENU2 mice showed increased mGlu5 receptor protein levels in the hippocampus and corpus striatum (but not in the prefrontal cortex) whereas the transcript of the mGlu5 receptor was unchanged. No changes in mGlu1 receptor mRNA and protein levels were found in any of the three brain regions of ENU2 mice. We extended the analysis to Homer proteins, which act as scaffolds by linking mGlu1 and mGlu5 receptors to effector proteins. Expression of the long isoforms of Homer was significantly reduced in the hippocampus of ENU2 mice, whereas levels of the short Homer isoform (Homer 1a) were unchanged. mGlu5 receptors were less associated to immunoprecipitated Homer in the hippocampus of ENU2 mice. The lack of mGlu5 receptor-mediated long-term depression (LTD) in wild-type mice (of BTBR strain) precluded the analysis of hippocampal synaptic plasticity in ENU2 mice. We therefore performed a behavioral analysis to examine whether pharmacological blockade of mGlu5 receptors could correct behavioral abnormalities in ENU2 mice. Using the same apparatus we sequentially assessed locomotor activity, object exploration, and spatial object recognition (spatial novelty test) after displacing some of the objects from their original position in the arena. Systemic treatment with the mGlu5 receptor antagonist, MPEP (20 mg/kg, i.p.), had a striking effect in the spatial novelty test by substantially increasing the time spent in exploring the displaced objects in ENU2 mice (but not in wild-type mice). These suggest a role for mGlu5 receptors in the pathophysiology of ID in PKU and suggest that, also in adult untreated animals, cognitive dysfunction may be improved by targeting these receptors with an appropriate therapy

Heat-shock pretreatment inhibits sorbitol-induced apoptosis in K562, U937 and HeLa cells.

The aim of this study was to determine whether heat-shock pretreatment exerted a protective effect against sorbitol-induced apoptotic cell death in K562, U937 and HeLa cell lines and whether such protection was associated with a decreased cytochrome c release from mithocondria and a decreased activation of caspase-9 and -3. Following heat-shock pretreatment (42 6 0.3C for 1 hr), these cell lines were exposed to sorbitol for 1 hr. Apoptosis was evaluated by DNA fragmentation, whereas caspase-9,-3 activation, cytochrome c release and heat-shock protein70 (HSP70) were assayed by Western Blot. Sorbitol exposure-induced apoptosis in these different cell lines with a marked activation of caspase-9 and caspase- 3, whereas heat-shock pretreatment before sorbitol exposure, induced expression of HSP70 and inhibited sorbitol-mediated cytochrome c release and subsequent activation of caspase-9 and caspase- 3. Similarly, overexpression of HSP70 in the three cell lines studied prevented caspase-9 cleavage and activation as well as cell death. Furthermore, we showed that the mRNA expression of iNOS decreased during both the heat-shock treatment and heat-shock pretreatment before sorbitol exposure. By contrast, the expression of Cu-Zn superoxide dismutase (SOD) and Mn-SOD proteins increased during heat-shock pretreatment before sorbitol exposure. We conclude that, heat-shock pretreatment protects different cell lines against sorbitol-induced apoptosis through a mechanism that is likely to involve SOD family members

Klippel-Feil syndrome. When using fiberoptic bronchoscopy guide, a case report

Klippel-Feil syndrome is a rare disease with congenital musculoskeletal condition characterized by faulty segmentation of cervical vertebrae and consists of cervical vertebra fusions with limitation of head movements, short neck and low posterior hairline. In several cases the syndrome is associated with cardiovascular malformations. Patients affected by Klippel-Feil syndrome could be an anesthetic challenge, not only during cardiac surgery. We are presenting a case of Klippel-Feil Syndrome in an adult patient, who was operated on for a pulmonary valve insufficiency in a previously corrected Tetralogy of Fallot Syndrome. We are going to discuss the features of this rare syndrome

Tidal signatures in Neogene to Quaternary mixed deposits of southern Italy straits and bays

Some of the Neogene to Quaternary sedimentary successions cropping out in the southern Italy orogenic belt exhibit distinct stratigraphic intervals of mixed, silici-bioclastic arenites. These deposits represent bay- and strait-fill successions that accumulated during tectonically-driven, rapid transgressions in peripheral marine basins of the central Mediterranean, experiencing microtidal conditions similar to those presently existing in the Mediterranean Sea. The Upper Miocene to Middle Pleistocene successions of Basilicata, Calabria and NE Sicily, show laterallyaccreted, cross-strata of mixed composition, with the siliciclastic fraction derived from either sedimentary or metamorphic rocks and the bioclastic fraction produced by an in situ or near situ heterozoan factory. Tidal cyclicity of semi-diurnal and diurnal to monthly and yearly periodicities has been detected in the studied deposits, where tidal bundling is revealed by the rhythmic alternation of siliciclastic and bioclastic set of laminae, repeated according to different cycles. This rhythmic signature appears to be more evident where randomly-occurring processes, such as waves, storms and currents, were mitigated by engulfed or strait palaeo-settings. Palaeo-bays preserved short-term tidal cycles in shoreface to offshore-transition mixed deposits because hydrodynamically isolated from open marine conditions and therefore subjected to tidal influence only during fair-weather periods. On the contrary, palaeo-straits recorded tidal cyclicities of longer duration in deeper mixed deposits subjected to steady tidal currents

The complex architecture of the 2009 MW 6.1 L’Aquila normal fault system (Central Italy) as imaged by 64,000 high-resolution aftershock locations.

On April 6th 2009, a MW 6.1 normal faulting earthquake struck the axial area of the Abruzzo region in Central Italy. We present high-precision hypocenter locations of an extraordinary dataset composed by 64,000 earthquakes recorded at a very dense seismic network of 60 stations operating for 9 months after the main event. Events span in magnitude (ML) between -0.9 to 5.9, reaching a completeness magnitude of 0.7. The dataset has been processed by integrating an accurate automatic picking procedure together with cross-correlation and double-difference relative location methods. The combined use of these procedures results in earthquake relative location uncertainties in the range of a few meters to tens of meters, comparable/lower than the spatial dimension of the earthquakes themselves). This data set allows us to image the complex inner geometry of individual faults from the kilometre to meter scale. The aftershock distribution illuminates the anatomy of the en-echelon fault system composed of two major faults. The mainshock breaks the entire upper crust from 10 km depth to the surface along a 14-km long normal fault. A second segment, located north of the normal fault and activated by two Mw>5 events, shows a striking listric geometry completely blind. We focus on the analysis of about 300 clusters of co-located events to characterize the mechanical behavior of the different portions of the fault system. The number of events in each cluster ranges from 4 to 24 events and they exhibit strongly correlated seismograms at common stations. They mostly occur where secondary structures join the main fault planes and along unfavorably oriented segments. Moreover, larger clusters nucleate on secondary faults located in the overlapping area between the two main segments, where the rate of earthquake production is very high with a long-lasting seismic decay

Interactions between the Multicolored Asian Lady Beetle Harmonia axyridis and the Parasitoid Dinocampus coccinellae

Harmonia axyridis (Pallas) has been introduced either intentionally or accidentally in different areas outside its native range, where it is often regarded as invasive. Dinocampus coccinellae (Schrank) has been recorded to parasitize H. axyridis in the field, both in the native and introduced areas, Italy included. The percent of parasitism found in our field investigation was low (four percent).The effect of exposure time of H. axyridis to D. coccinellae and the impact of parasitization on host longevity, oviposition capacity and egg fertility were evaluated in the laboratory. The acceptance and suitability of H. axyridis as host for D. coccinellae were then studied, in comparison with the native coccinellid Adalia bipunctata (L.), which shares the same ecological niche. The effects of parasitization on female longevity and reproduction capacity in the exotic vs. the indigenous lady beetle were also investigated. The overall results showed that D. coccinellae negatively affected the fitness of H. axyridis, more than that of A. bipunctata. The parasitoid may thus play a marginal role in controlling the populations of the Asian lady beetle, without representing a threat to A. bipunctata

E2F and p53 Induce Apoptosis Independently during Drosophila Development but Intersect in the Context of DNA Damage

In mammalian cells, RB/E2F and p53 are intimately connected, and crosstalk between these pathways is critical for the induction of cell cycle arrest or cell death in response to cellular stresses. Here we have investigated the genetic interactions between RBF/E2F and p53 pathways during Drosophila development. Unexpectedly, we find that the pro-apoptotic activities of E2F and p53 are independent of one another when examined in the context of Drosophila development: apoptosis induced by the deregulation of dE2F1, or by the overexpression of dE2F1, is unaffected by the elimination of dp53; conversely, dp53-induced phenotypes are unaffected by the elimination of dE2F activity. However, dE2F and dp53 converge in the context of a DNA damage response. Both dE2F1/dDP and dp53 are required for DNA damage-induced cell death, and the analysis of rbf1 mutant eye discs indicates that dE2F1/dDP and dp53 cooperatively promote cell death in irradiated discs. In this context, the further deregulation in the expression of pro-apoptotic genes generates an additional sensitivity to apoptosis that requires both dE2F/dDP and dp53 activity. This sensitivity differs from DNA damage-induced apoptosis in wild-type discs (and from dE2F/dDP-induced apoptosis in un-irradiated rbf1 mutant eye discs) by being dependent on both hid and reaper. These results show that pro-apoptotic activities of dE2F1 and dp53 are surprisingly separable: dp53 is required for dE2F-dependent apoptosis in the response to DNA damage, but it is not required for dE2F-dependent apoptosis caused simply by the inactivation of rbf1

Reproductive activity in sheep with different lambing period treated with melatonin in April

The object was to evaluate the effect of melatonin treatment on the advance in April of the reproductive resumption in Sarda breed sheep with different lambing period. For the research two farms, located in North Sardinia between 39° and 40° N, were chosen. In each farm, 120 lactating ewes were selected: 30 lambed between October 20th and November 20th (group 1); 30 lambed between December 1st and 30th (group 2); 30 lambed between January 1st and 30th; 30 lambed between February 1st and 28th (group 4). In each farm, each group of 30 animals was divided into two subgroups of 15 animals (M and C). On April 1st, in each farm, the animals of the M subgroups were treated with a implant containing 18 mg melatonin. The subgroups C were kept as control. The lambing dates and the number of newborn lambs were recorded until 220 days after ram introduction. In treated animals greatest fertility (P<0.01) and lowest distance in days from male introduction to lambing (P<0.01) were recorded. The best reproductive performances were found in the group 1 and 2 compared to the other two groups (P<0.01).The present research shows that melatonin treatment should be made 3 or 4 months after lambing, in order to obtain optimal results

Characterization of E2F8, a novel E2F-like cell-cycle regulated repressor of E2F-activated transcription

The E2F family of transcription factors are downstream effectors of the retinoblastoma protein, pRB, pathway and are essential for the timely regulation of genes necessary for cell-cycle progression. Here we describe the characterization of human and murine E2F8, a new member of the E2F family. Sequence analysis of E2F8 predicts the presence of two distinct E2F-related DNA binding domains suggesting that E2F8 and, the recently, identified E2F7 form a subgroup within the E2F family. We show that E2F transcription factors bind the E2F8 promoter in vivo and that expression of E2F8 is being induced at the G1/S transition. Purified recombinant E2F8 binds specifically to a consensus E2F-DNA-binding site indicating that E2F8, like E2F7, binds DNA without the requirement of co-factors such as DP1. E2F8 inhibits E2F-driven promoters suggesting that E2F8 is transcriptional repressor like E2F7. Ectopic expression of E2F8 in diploid human fibroblasts reduces expression of E2F-target genes and inhibits cell growth consistent with a role for repressing E2F transcriptional activity. Taken together, these data suggest that E2F8 has an important role in turning of the expression of E2F-target genes in the S-phase of the cell cycle