Peripheral neuropathy is characterized by abnormal pain responses triggered by the release of several mediators and neuronal hyperexcitability at the spinal cord level. Emerging evidence indicates ...that the enhanced activity of dorsal horn neurons requires communication with glia and microglia, cells that are physiologically involved in neuronal wellbeing. Prokineticins (PKs), which include PK1 and PK2, represent a novel family of chemokines characterized by a unique structural motif comprising five disulfide bonds. They are expressed in the peripheral and central nervous system. PKs bind two G protein coupled receptors, PKR1 and PKR2, and participate in the regulation of several biological processes, including pain sensation. This study aimed to investigate the anti-nociceptive effect of PC1, a non-peptide PKR1-preferring antagonist, in a mouse model of neuropathic pain. To do this, we assessed the activity of spinal cord nociceptive neurons as well as astrocyte and microglia phenotypes after repeated administration of PC1 in vivo. PC1 treatment strongly delayed the development of thermal hyperalgesia and tactile and mechanical allodynia. It also reduced spinal microglial and glial activation 8 days post injury in spared nerve injury (SNI) mice. Neuropathic mice showed an increased level of PK2 protein in the spinal cord, mostly in astrocytes. PC1 treatment completely reversed the increased responsiveness to mechanical stimuli, the decreased threshold of neuronal activation, and the increased spontaneous activity that were observed in nociceptive specific (NS) neurons of SNI mice.
Objective: Global Maxwell Tomography (GMT) is a recently introduced volumetric technique for noninvasive estimation of electrical properties (EP) from magnetic resonance measurements. Previous work ...evaluated GMT using ideal radiofrequency (RF) excitations. The aim of this simulation study was to assess GMT performance with a realistic RF coil. Methods: We designed a transmit-receive RF coil with 8 decoupled channels for 7T head imaging. We calculated the RF transmit field (<inline-formula><tex-math notation="LaTeX">B_1^+</tex-math></inline-formula>) inside heterogeneous head models for different RF shimming approaches, and used them as input for GMT to reconstruct EP for all voxels. Results: Coil tuning/decoupling remained relatively stable when the coil was loaded with different head models. Mean error in EP estimation changed from <inline-formula><tex-math notation="LaTeX">\text{7.5}{\%}</tex-math></inline-formula> to <inline-formula><tex-math notation="LaTeX">\text{9.5}{\%}</tex-math></inline-formula> and from <inline-formula><tex-math notation="LaTeX">\text{4.8}{\%}</tex-math></inline-formula> to <inline-formula><tex-math notation="LaTeX">\text{7.2}\%</tex-math></inline-formula> for relative permittivity and conductivity, respectively, when changing head model without re-tuning the coil. Results slightly improved when an SVD-based RF shimming algorithm was applied, in place of excitation with one coil at a time. Despite errors in EP, RF transmit field (<inline-formula><tex-math notation="LaTeX">B_1^+</tex-math></inline-formula>) and absorbed power could be predicted with less than <inline-formula><tex-math notation="LaTeX">\text{0.5}{\%}</tex-math></inline-formula> error over the entire head. GMT could accurately detect a numerically inserted tumor. Conclusion: This work demonstrates that GMT can reliably reconstruct EP in realistic simulated scenarios using a tailored 8-channel RF coil design at 7T. Future work will focus on construction of the coil and optimization of GMT's robustness to noise, to enable in-vivo GMT experiments. Significance: GMT could provide accurate estimations of tissue EP, which could be used as biomarkers and could enable patient-specific estimation of RF power deposition, which is an unsolved problem for ultra-high-field magnetic resonance imaging.
Craniosynostosis (CS), the premature fusion of one or more cranial sutures, is a relatively common congenital anomaly, occurring in 3–5 per 10,000 live births. Nonsyndromic CS (NCS) accounts for up ...to 80% of all CS cases, yet the genetic factors contributing to the disorder remain largely unknown. The RUNX2 gene, encoding a transcription factor critical for bone and skull development, is a well known CS candidate gene, as copy number variations of this gene locus have been found in patients with syndromic craniosynostosis. In the present study, we aimed to characterize RUNX2 to better understand its role in the genetic etiology and in the molecular mechanisms underlying midline suture ossification in NCS. We report four nonsynonymous variants, one intronic variant and one 18 bp in-frame deletion in RUNX2 not found in our study control population. Significant difference in allele frequency (AF) for the deletion variant RUNX2 p.Ala84-Ala89del (ClinVar 257,095; dbSNP rs11498192) was observed in our sagittal NCS cohort when compared to the general population (P = 1.28 × 10−6), suggesting a possible role in the etiology of NCS. Dual-luciferase assays showed that three of four tested RUNX2 variants conferred a gain-of-function effect on RUNX2, further suggesting their putative pathogenicity in the tested NCS cases. Downregulation of RUNX2 expression was observed in prematurely ossified midline sutures. Metopic sites showed significant downregulation of promoter 1-specific isoforms compared to sagittal sites. Suture-derived mesenchymal stromal cells showed an increased expression of RUNX2 over matched unfused suture derived cells. This demonstrates that RUNX2, and particularly the distal promoter 1-isoform group, are overexpressed in the osteogenic precursors within the pathological suture sites.
•RUNX2 encodes the master bone transcription factor critical for skull development.•Rare RUNX2 variants may contribute to the pathogenesis of midline nonsyndromic CS.•A RUNX2 p.Ala84-Ala89del variant was significantly enriched in sagittal CS.•RUNX2 transcripts with the P1 promoter are overexpressed in fused sutures of CS patients.
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•The Montevecchio-Ingurtosu district was one of the largest Italian Pb-Zn-Ag producers.•New field, microscopic, microchemical, isotopic, and fluid inclusion data were obtained.•The ...Montevecchio deposit shows several affinities with the class of Ag-Pb-Zn veins in metasediments as defined by Beaudoin & Sangster (1992).•Ore textures and mineralogical-chemical features at all scales document a multistage process of mineralization postdating the emplacement of the 304 ± 4 Ma Arbus pluton, and occurring in a tectonically active regime.•The exact age of mineralization is not constrained, although it might be related to regional, large-scale shear deformation in a late-post-orogenic context.•At least part of the mineralization is associated with relatively low temperature (≦200 °C), high salinity (>20 wt% equiv. mass NaCl) fluids analogous to basinal brines.
The Montevecchio-Ingurtosu district (SW Sardinia) was among the largest historical Pb-Zn producers of Italy, with significant Ag output, and yields of metallic byproducts such as Cd, Co, Cu, Bi, Sb, Ge, In, and Ga. Despite this importance, detailed descriptions of orebodies and mineral assemblages are surprisingly scarce and old. Here we summarize all previously available information and contribute new textural, microchemical, fluid inclusion, and C-O isotopic data deriving from a recent sampling of currently exposed orebodies. Mineralization occurs as steeply dipping veins extending for almost 20 km at the northern and western flanks of the Variscan Arbus pluton; veins are hosted in phyllites of the Arburese Unit close to the thermometamorphic aureole. Ore enrichments range from massive to coarsely nodular, stockwork and brecciated. Vein filling consists of siderite (locally Zn-rich), quartz, sphalerite, galena (remarkably rich in micro-inclusions), with accessory chalcopyrite, fahlore, bournonite, barite, and Ni-Co sulfarsenides. Vein assemblages may vary, even within a single vein, from sphalerite- to galena-dominated. Cu-rich assemblages (fahlore + chalcopyrite) were locally observed. Wallrock is affected by silicification, sericitization and pyrite dissemination. Sphalerite shows a complex mineral chemistry, with marked variations in Fe (0.08 to 8.9 mol% FeS) and in Cd contents (from <0.05 to >1 wt%). Trace element analyses (LA-ICP-MS) document the presence of measurable amounts of Ga, Ge, In, Ag, Ni, Co, Tl and Mo. Fahlore varies from the Sb (tetrahedrite) and the As (tennantite) endmembers, with Ag-rich compositions corresponding to Sb endmembers. The occurrence of Cd-rich tetrahedrite (up to 14.5 wt%) is remarkable. Fluid inclusion studies on sphalerite and quartz are in agreement with previous literature data, and indicate comparatively low (in the range 90°–130 °C) homogenization temperatures (Th) and high salinities (14 to 25 equiv. mass % NaCl); both microthermometric data and SEM/EDS analyses of decrepitated inclusions indicate the presence of Ca and K beside Na. Pressure correction to Th is presumably low (in the order of 10 °C), considering a hydrostatic pressure regime. Application of the sphalerite-based GGIMFiS thermometer provided temperatures in excellent agreement with fluid inclusion data for the Montevecchio mineralization. Isotopic data on Montevecchio carbonates define mildy negative δ13CPDB values (between −1 and −5 per mil), and a larger spread of δ18OSMOW values, between +14 and +20 per mil. Other isotopic data from literature include a limited set of δ34S values for galena, sphalerite and pyrite, and a fairly large body of Pb isotopic data on galena. Comparatively narrow ranges for sulfur isotope signatures suggest a homogeneous, possibly igneous, sulfur source, whereas the large spread in Pb isotope signatures reflects the large-scale character of the hydrothermal process and indicates that the Arbus pluton may have been just one of the sources of ore lead. The Montevecchio vein system shows affinities with other late to post Variscan European deposits. The nature of the mineralizing fluids at Montevecchio is analogous to low temperature, basinal brines, and typical of several regional-scale, late to post-Variscan hydrothermal events across Europe. Mineralization at Montevecchio obviously postdates the emplacement of the Arbus plutonic complex, but its exact age remains unknown.
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Despite its documented prognostic relevance, hepatic encephalopathy (HE) is not considered in liver transplantation (LT) due to its possible poor objectivity. To override this problem, we aimed to ...analyze if an objective diagnosis of HE may confer additional mortality risk beyond MELD. Study and validation cohorts of patients with cirrhosis were considered in Italy and Canada, respectively. Patients were considered to be HE+ if an episode of overt HE was documented in a hospitalization. Of the 486 patients enrolled in Italy, 184 (38%) were HE+. During the 6‐month follow‐up, 77 patients died and 50 underwent transplantation. The 6‐month mortality of HE+ versus HE– patients was significantly higher (P < 0.001). Model for End‐Stage Liver Disease (MELD; subdistribution hazard ratio sHR, 1.2; 95% confidence interval CI, 1.1‐1.2; P < 0.001), HE+ (sHR, 3.6; 95% CI, 1.8‐7.1; P < 0.001), and sodium (sHR, 0.9; 95% CI, 0.8‐0.9; P < 0.001) were independent predictors of 6‐month mortality. In HE+ patients, short‐term mortality increased across the entire MELD spectrum (range, 6‐40). The results were unchanged by including or excluding patients with hepatocellular carcinoma or stratifying patients according to HE characteristics. The higher 6‐month mortality of HE+ versus HE– patients was confirmed also in the Canadian cohort (P < 0.001; n = 300, 33% HE+; 33 died, 104 transplanted). A similar and statistically significant C‐index increase derived by the incorporation of HE in MELD was observed both in the Italian (from 0.67 to 0.75) and Canadian (from 0.69 to 0.74) cohorts. A score based on MELD plus 7 points (95% CI, 4‐10) for HE+ patients optimally predicted 6‐month mortality in the 2 cohorts. According to the net reclassification index, by not considering HE, 29% of overall patients were misclassified by MELD score. In conclusion, the incorporation of HE in MELD score might improve the listing and allocation policy in LT. Liver Transplantation 22 1333–1342 2016 AASLD.
Background & Aims
Despite the excellent efficacy of direct‐acting antivirals (DAA) reported in clinical trials, virological failures can occur, often associated with the development of ...resistance‐associated substitutions (RASs). This study aimed to characterize the presence of clinically relevant RASs to all classes in real‐life DAA failures.
Methods
Of the 200 virological failures that were analyzed in 197 DAA‐treated patients, 89 with pegylated‐interferon+ribavirin (PegIFN+RBV) and 111 without (HCV‐1a/1b/1g/2/3/4=58/83/1/6/24/25; 56.8% treatment experienced; 65.5% cirrhotic) were observed. Sanger sequencing of NS3/NS5A/NS5B was performed by home‐made protocols, at failure (N=200) and whenever possible at baseline (N=70).
Results
The majority of the virological failures were relapsers (57.0%), 22.5% breakthroughs, 20.5% non‐responders. RAS prevalence varied according to IFN/RBV use, DAA class, failure type and HCV genotype/subtype. It was 73.0% in IFN group vs 49.5% in IFN free, with the highest prevalence of NS5A‐RASs (96.1%), compared to NS3‐RASs (75.9% with IFN, 70.5% without) and NS5B‐RASs (66.6% with IFN, 20.4% without, in sofosbuvir failures). In the IFN‐free group, RASs were higher in breakthrough/non‐responders than in relapsers (90.5% vs 40.0%, P<.001). Interestingly, 57.1% of DAA IFN‐free non‐responders had a misclassified genotype, and 3/4 sofosbuvir breakthroughs showed the major‐RAS‐S282T, while RAS‐L159F was frequently found in sofosbuvir relapsers (18.2%). Notably, 9.0% of patients showed also extra target RASs, and 47.4% of patients treated with ≥2 DAA classes showed multiclass resistance, including 11/11 NS3+NS5A failures. Furthermore, 20.0% of patients had baseline‐RASs, which were always confirmed at failure.
Conclusions
In our failure setting, RAS prevalence was remarkably high in all genes, with a partial exception for NS5B, whose limited resistance is still higher than previously reported. This multiclass resistance advocates for HCV resistance testing at failure, in all three genes for the best second‐line therapeutic tailoring.
See Editorial on Page 506
This work provides an overview of plant diversity in the municipality of Rome (Italy) through an assessment of the flora in urban and suburban sectors of the city. It is aimed at providing the ...knowledge required to support proactive action for plant conservation. On the basis of a literature-derived catalogue and of an extensive survey campaign, the flora was investigated in terms of conservation interest; habitat types and locations that require protection measures were identified according to the occurrence of valuable native plants. Valuable species exclusive of the urban sector are threatened by the compaction trend affecting the urban fabric. Such species occur mainly in ruderal environments and fallows of archaeological sites and urban parks, near river courses and in remnants of natural forests. Valuable species exclusive of the suburban sector are threatened by urban sprawl. They occur prevalently in wet environments of the subcoastal strip and in coastal sands, Mediterranean maquis, tuffaceous gorges, sulphur springs and archaeological sites. The results highlight the need to preserve the complexity of the land mosaic, especially within the urban matrix, and to strengthen the existing environmental protection tools in the suburban area against foreseeable land cover changes.
Enhancing pasture persistence is crucial to achieve more sustainable grass‐based animal production systems. Although it is known that persistence of perennial ryegrass is based on a high turnover of ...tillers during late spring and summer, little is known about other forage species, particularly in subtropical climates. To address this question, this study evaluated survival of grazed tall fescue tillers growing in a subtropical climate. We hypothesized that hard tactical grazing during winter to remove reproductive stems (designated as ‘flowering control’), and nitrogen fertilization in spring, would both improve tiller survival over summer, and thus enhance tiller density. This was assessed in two experiments. In both experiments, few tillers appeared during late spring and summer and so tiller density depended on the dynamics of vegetative tillers present in the sward in spring. In Experiment 2, flowering control and nitrogen fertilization both enhanced the survival of that critical tiller cohort, but the effects were not additive. Responses were similar but not statistically significant in Experiment 1, which had a warmer, drier summer and lower overall survival rates. Unlike grasses in temperate environments, persistence of tall fescue in this subtropical site appeared to follow a ‘vegetative pathway’; i.e., new tillers were produced largely in autumn, from vegetative tillers that survived the summer.
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