Modification of Cul1 regulates its association with proteasomal subunits

BACKGROUND: Ubiquitylation targets proteins for degradation by the 26S proteasome. Some yeast and plant ubiquitin ligases, including the highly conserved SCF (Skp1/Cul1/F-box protein) complex, have been shown to associate with proteasomes. We sought to characterize interactions between SCF complexes and proteasomes in mammalian cells. RESULTS: We found that the binding of SCF complexes to proteasomes is conserved in higher eukaryotes. The Cul1 subunit associated with both sub-complexes of the proteasome, and high molecular weight forms of Cul1 bound to the 19S proteasome. Cul1 is ubiquitylated in vivo. Ubiquitylation of Cul1 promotes its binding to the S5a subunit of the 19S sub-complex without affecting Cul1 stability. CONCLUSION: The association of ubiquitylating enzymes with proteasomes may be an additional means to target ubiquitylated substrates for degradation

Effect of Non-psychotropic Plant-derived Cannabinoids on Bladder Contractility: Focus on Cannabigerol.

There are anecdotal reports that some Cannabis preparations may be useful for bladder dysfunctions. Here, we investigated the effect of a number of non-psychotropic phytocannabinoids, namely cannabidiol (CBD), cannabigerol (CBG), cannabidivarin (CBDV), Δ9-tetrahydrocannabivarin (THCV) and cannabichromene (CBC) on mouse bladder contractility in vitro. CBG, THCV, CBD and CBDV, but not CBC, at concentration ranging from 10−8M to 10−4M, decreased (with similar potency), the contractions induced by acetylcholine without significantly modifying the contractions induced by electrical stimulation. The rank order of efficacy was CBG=THCV>CBD>CBDV. In depth studies on CBG showed that the effect of this phytocannabinoid on acetylcholine-induced contractions was not affected by CB1or CB2receptor antagonists. Additionally, CBG also reduced acetylcholine-induced contractions in the human bladder

Unraveling the CMB lack-of-correlation anomaly with the cosmological gravitational wave background

Since the very first observations, the Cosmic Microwave Background (CMB) has revealed on large-scales unexpected features known as anomalies, which challenge the standard $\Lambda$ cold dark matter ($\Lambda$CDM) cosmological model. One such anomaly is the ``lack-of-correlation'', where the measured two-point angular correlation function of CMB temperature anisotropies is compatible with zero, differently from the predictions of the standard model. This anomaly could indicate a deviation from the standard model, unknown systematics, or simply a rare realization of the model itself. In this study, we explore the possibility that the lack-of-correlation anomaly is a consequence of living in a rare realization of the standard model, by leveraging the potential information provided by the cosmological gravitational wave background (CGWB) detectable by future gravitational wave (GW) interferometers. We analyze both constrained and unconstrained realizations of the CGWB to investigate the extent of information that GWs can offer. To quantify the impact of the CGWB on the lack-of-correlation anomaly, we employ established estimators and introduce a new estimator that addresses the ``look-elsewhere'' effect. Additionally, we consider three different maximum multipoles, denoted as $\ell_{\rm max}$, to account for the anticipated capabilities of future GW detectors ($\ell_{\rm max} = 4, 6, 10$). Summarizing our findings for the case of $\ell_{\rm max} = 4$, we identify the angular range $[63^\circ - 180^\circ]$ as the region where future observations of the CGWB maximize the probability of rejecting the standard model. Furthermore, we calculate the expected significance of this observation, demonstrating that 98.81% (81.67%) of the GW realizations enhance the current significance of the anomaly when considering the full-sky (masked) Planck SMICA map as our CMB sky

The addition of simvastatin administration to cold storage solution of explanted whole liver grafts for facing ischemia/reperfusion injury in an area with a low rate of deceased donation: a monocentric randomized controlled double-blinded phase 2 study.

BACKGROUND: Liver transplantation is the best treatment for end-stage liver disease. The interruption of the blood supply to the donor liver during cold storage damages the liver, affecting how well the liver will function after transplant. The drug Simvastatin may help to protect donor livers against this damage and improve outcomes for transplant recipients. The aim of this study is to evaluate the benefits of treating the donor liver with Simvastatin compared with the standard transplant procedure. PATIENT AND METHODS: We propose a prospective, double-blinded, randomized phase 2 study of 2 parallel groups of eligible adult patients. We will compare 3-month, 6-month, and 12-month graft survival after LT, in order to identify a significant relation between the two homogenous groups of LT patients. The two groups only differ by the Simvastatin or placebo administration regimen while following the same procedure, with identical surgical instruments, and medical and nursing skilled staff. To reach these goals, we determined that we needed to recruit 106 patients. This sample size achieves 90% power to detect a difference of 14.6% between the two groups survival using a one-sided binomial test. DISCUSSION: This trial is designed to confirm the effectiveness of Simvastatin to protect healthy and steatotic livers undergoing cold storage and warm reperfusion before transplantation and to evaluate if the addition of Simvastatin translates into improved graft outcomes. TRIAL REGISTRATION: ISRCTN27083228

Damage patterns in the town of Amatrice after August 24th 2016 Central Italy earthquakes

The impact of the two seismic events of August 24th 2016 on the municipality of Amatrice was highly destructive. There were 298 victims, 386 injured, about 5000 homeless, and the historical center of the town suffered a great number of partial and total collapses. The 260 strong motion records obtained for the first event were analyzed and plotted in a shakemap, comparing them with the macroseismic damage surveys made in 305 localities. On the basis of an inspection survey made in September 2016, a map of the damage patterns of the buildings in the historical center was elaborated according to the EMS 98 classification. The damage level resulted very high with more than 60% of the inspected buildings showing partial or total collapse. The elevated level of destruction was mainly caused by the high vulnerability of the masonry buildings, mostly due to specific vulnerability factors such as the poor quality of masonry, the lack of connections between walls and the poor connection between external walls and floors

Circumferential dissection of deep fascia as ancillary technique in circumcision: is it possible to correct phimosis increasing penis size?

Background: Phimosis is the inability to retract the preputium downward over the glans penis. Despite the various techniques of preputial plasty described in literature, the most performed surgical treatment is still the conventional circumcision.Methods: In this paper we retrospectively reviewed data of a homogeneous population of 36 consecutive adult patients who underwent phimosis correction by circumcsion with dissection of the Deep Fascia. Patients were followed up by one independent plastic surgeon that measured penis length and circumference in nonerected state preoperatively and at 6 month time postoperatively.Results: The Wilcoxon Signed Rank Test showed a significant (p < 0.0001) difference between the two groups both in terms of length and circumference.Conclusions: In conclusion, the ancillary technique we described leads to an increase of penis size, is safe and easy to perform and does not increase significantly operative time nor complication rate to the conventional procedure

Understanding the molecular basis of folding cooperativity through a comparative analysis of a multidomain protein and its isolated domains

Although cooperativity is a well-established and general property of folding, our current understanding of this feature in multi-domain folding is still relatively limited. In fact, there are contrasting results indicating that the constituent domains of a multi-domain protein may either fold independently on each other or exhibit inter-dependent supradomain phenomena. To address this issue, here we present the comparative analysis of the folding of a tandem repeat protein, comprising two contiguous PDZ domains, in comparison to that of its isolated constituent domains. By analyzing in detail the equilibrium and kinetics of folding at different experimental conditions, we demonstrate that, despite each of the PDZ domains in isolation being capable of independent folding, at variance with previously characterized PDZ tandem repeats, the full-length construct folds and unfolds as a single co-operative unit. By exploiting quantitatively the comparison of the folding of the tandem repeat to those observed for its constituent domains, as well as by characterizing a truncated variant lacking a short auto-inhibitory segment, we successfully rationalize the molecular basis of the observed cooperativity and attempt to infer some general conclusions for multi-domain systems

Folding and Binding Mechanisms of the SH2 Domain from Crkl

SH2 domains are structural modules specialized in the recognition and binding of target sequences containing a phosphorylated tyrosine residue. They are mostly incorporated in the 3D structure of scaffolding proteins that represent fundamental regulators of several signaling pathways. Among those, Crkl plays key roles in cell physiology by mediating signals from a wide range of stimuli, and its overexpression is associated with several types of cancers. In myeloid cells expressing the oncogene BCR/ABL, one interactor of Crkl-SH2 is the focal adhesion protein Paxillin, and this interaction is crucial in leukemic transformation. In this work, we analyze both the folding pathway of Crkl-SH2 and its binding reaction with a peptide mimicking Paxillin, under different ionic strength and pH conditions, by using means of fluorescence spectroscopy. From a folding perspective, we demonstrate the presence of an intermediate along the reaction. Moreover, we underline the importance of the electrostatic interactions in the early event of recognition, occurring between the phosphorylated tyrosine of the Paxillin peptide and the charge residues of Crkl-SH2. Finally, we highlight a pivotal role of a highly conserved histidine residue in the stabilization of the binding complex. The experimental results are discussed in light of previous works on other SH2 domains

Magnetotransport and magnetic properties of amorphous NdNi 5 thin films

Abstract: NdNi5 is an intermetallic compound with a bulk Curie temperature (TCurie) of 6–13 K. While existing studies have focused on NdNi5 crystals, amorphous thin-films of NdNi5 are potentially important since they would be magnetically soft without magnetocrystalline anisotropy, meaning that small external magnetic fields could reverse the direction of their magnetization. Here, we report NdNi5 thin-films with a thickness in the 5–200 nm range, deposited by DC magnetron sputtering onto Si(100). Films are amorphous with a weak temperature-dependent resistivity with values ranging between 150 and 300 μΩ cm. By means of noise spectroscopy, by analyzing the time-dependence of fluctuation-induced voltages, it is found that at low temperatures the resistance fluctuations are due to the Kondo effect. Volume magnetometry indicates TCurie=70 K with a magnetic coercive field of 30 mT at 5 K for a 125-nm-thick film. The results are promising for the development of Ferromagnet(F)/Superconductor(S)/Ferromagnet(F) pseudo spin-valve devices based on amorphous NdNi5 thin films

The Cdc14B-Cdh1-Plk1 Axis Controls the G2 DNA-Damage-Response Checkpoint

n response to DNA damage in G2, mammalian cells must avoid entry into mitosis and instead initiate DNA repair. Here, we show that, in response to genotoxic stress in G2, the phosphatase Cdc14B translocates from the nucleolus to the nucleoplasm and induces the activation of the ubiquitin ligase APC/CCdh1, with the consequent degradation of Plk1, a prominent mitotic kinase. This process induces the stabilization of Claspin, an activator of the DNA-damage check- point, and Wee1, an inhibitor of cell-cycle progres- sion, and allows an efficient G2 checkpoint. As a by-product of APC/CCdh1 reactivation in DNA-dam- aged G2 cells, Claspin, which we show to be an APC/ CCdh1 substrate in G1, is targeted for degradation. However, this process is counteracted by the deubi- quitylating enzyme Usp28 to permit Claspin-medi- ated activation of Chk1 in response to DNA damage. These findings define a novel pathway that is crucial for the G2 DNA-damage-response checkpoint