Phospholipase C Gamma 2 Is Critical for Development of a Murine Model of Inflammatory Arthritis by Affecting Actin Dynamics in Dendritic Cells

BACKGROUND:Dendritic cells (DCs) are highly specialized cells, which capture antigen in peripheral tissues and migrate to lymph nodes, where they dynamically interact with and activate T cells. Both migration and formation of DC-T cell contacts depend on cytoskeleton plasticity. However, the molecular bases governing these events have not been completely defined. METHODOLOGY/PRINCIPAL FINDINGS:Utilizing a T cell-dependent model of arthritis, we find that PLCgamma2-/- mice are protected from local inflammation and bone erosion. PLCgamma2 controls actin remodeling in dendritic cells, thereby affecting their capacity to prime T cells. DCs from PLCgamma2-/- mice mature normally, however they lack podosomes, typical actin structures of motile cells. Absence of PLCgamma2 impacts both DC trafficking to the lymph nodes and migration towards CCL21. The interaction with T cells is also affected by PLCgamma2 deficiency. Mechanistically, PLCgamma2 is activated by CCL21 and modulates Rac activation. Rac1/2-/- DCs also lack podosomes and do not respond to CCL21. Finally, antigen pulsed PLCgamma2-/- DCs fail to promote T cell activation and induce inflammation in vivo when injected into WT mice. Conversely, injection of WT DCs into PLCgamma2-/- mice rescues the inflammatory response but not focal osteolysis, confirming the importance of PLCgamma2 both in immune and bone systems. CONCLUSIONS/SIGNIFICANCE:This study demonstrates a critical role for PLCgamma2 in eliciting inflammatory responses by regulating actin dynamics in DCs and positions the PLCgamma2 pathway as a common orchestrator of bone and immune cell functions during arthritis

Validation of the ONKOTEV Risk Prediction Model for Venous Thromboembolism in Outpatients With Cancer

Importance: The assessment of the risk of venous thromboembolism (VTE) among outpatients with cancer represents an unsolved topic. Current international guidelines recommend primary prophylaxis for patients at intermediate to high risk of VTE, indicated by a Khorana score of 2 or more. A previous prospective study developed the ONKOTEV score, a 4-variable risk assessment model (RAM) consisting of a Khorana score of more than 2, metastatic disease, vascular or lymphatic compression, and previous VTE event. Objective: To validate the ONKOTEV score as a novel RAM to assess the risk of VTE among outpatients with cancer. Design, setting, and participants: ONKOTEV-2 is a noninterventional prognostic study conducted in 3 European centers located in Italy, Germany, and the United Kingdom among a prospective cohort of 425 ambulatory patients with a histologically confirmed diagnosis of a solid tumor who were receiving active treatments. The total study duration was 52 months, with an accrual period of 28 months (from May 1, 2015, to September 30, 2017) and an overall follow up-period of 24 months (data were censored September 30, 2019). Statistical analysis was performed in October 2019. Exposures: The ONKOTEV score was calculated for each patient at baseline by collecting clinical, laboratory, and imaging data from tests performed for routine practice. Each patient was then observed to detect any thromboembolic event throughout the study period. Main outcomes and measures: The primary outcome of the study was the incidence of VTE, including deep vein thrombosis and pulmonary embolism. Results: A total of 425 patients (242 women [56.9%]; median age, 61 years [range, 20-92 years]) were included in the validation cohort of the study. The cumulative incidences for the risk of developing VTE at 6 months were 2.6% (95% CI, 0.7%-6.9%), 9.1% (95% CI, 5.8%-13.2%), 32.3% (95% CI, 21.0%-44.1%), and 19.3% (95% CI, 2.5%-48.0%), respectively, among 425 patients with an ONKOTEV score of 0, 1, 2, and greater than 2 (P < .001). The time-dependent area under the curve at 3, 6, and 12 months was 70.1% (95% CI, 62.1%-78.7%), 72.9% (95% CI, 65.6%-79.1%), and 72.2% (95% CI, 65.2%-77.3%), respectively. Conclusions and relevance: This study suggests that, because the ONKOTEV score has been validated in this independent study population as a novel predictive RAM for cancer-associated thrombosis, it can be adopted into practice and into clinical interventional trials as a decision-making tool for primary prophylaxis

Resources for assessing parents’ vaccine hesitancy: a systematic review of the literature

Vaccine hesitancy (VH) is a complex and country-specific issue, responsible for the decreasing vaccination rate and subsequent spread of vaccine-preventable diseases. In literature, several questionnaires were developed to assess VH. The aim of this systematic review was to evaluate the published questionnaires assessing parental VH. The search was conducted in PubMed/Medline, Web of Science and The Cochrane Library, in December 2017, following the PRISMA guidelines. The search strategy included 4 types of keywords: parents, vaccine hesitancy/acceptance, immunization and survey. Only English and Italian original papers were included. 17 reviewers independently screened titles and abstracts. Only the included articles were downloaded in full and, after a second screening, data were extracted and recorded in an ad hoc spreadsheet. A total of 5,139 articles were retrieved, after duplicates elimination 3,508 papers were screened. After a screening selection, 334 studies were included in the analysis. Most studies were cross-sectional (92.8%), followed by case-control (4.8%) and cohort studies (2.4%). The population interviewed was mainly parents, without any further details (73.1%); mothers were the only parent surveyed in approximately 20% of the studies, while only 1 study involved selectively the fathers. The sample size ranged from 7 to 59,897. Only 38% of the included studies reported both the number and type of items used. Regarding the type, more than half consisted of closed questions, followed by Likert scales, while open-ended questions were used in 14.8% of the surveys. Frequently, the survey was conducted using a self-reported questionnaire or interview. The questionnaires were mostly administered on paper, while online forms were used in 20.1% of the cases. However, 80.2% of the questionnaires were not attached to the paper. HPV vaccine was the most frequently investigated (39.2%), followed by influenza (13.5%) and measles (10.8%). While 22.4% of the articles referred to paediatrics vaccinations in general. Data about the immunization behaviours were reported in 294 studies: the subjects involved showed a behaviour defined as “acceptance” in 129 studies (38.6%), as “hesitancy/scepticism/doubt” in 145 studies (43.1%) and as “refusal” in 22 studies (6.6%). This information was not reported in 12% of the studies. VH is still a public health challenge, as confirmed by the high number of studies and questionnaires retrieved. This study offers a deeper perspec- tive on the available questionnaires, helping to identify the best one in terms of aim and study setting. &nbsp; &nbsp; &nbsp

Supplement: "Localization and broadband follow-up of the gravitational-wave transient GW150914" (2016, ApJL, 826, L13)

This Supplement provides supporting material for Abbott et al. (2016a). We briefly summarize past electromagnetic (EM) follow-up efforts as well as the organization and policy of the current EM follow-up program. We compare the four probability sky maps produced for the gravitational-wave transient GW150914, and provide additional details of the EM follow-up observations that were performed in the different bands

"That was the Elixir, the Stone, the Gold". Alchimia e individuazione del Sé in The Chymical Wedding di L. Clarke.

Questa ricerca ha avuto per oggetto l’analisi del romanzo The Chymical Wedding (1989) di Lindsay Clarke. Questo romanzo, apprezzato soprattutto in territorio anglosassone, è ancora poco conosciuto in Italia né è stato tradotto, fino ad oggi. L’ipotesi interpretativa da cui parte la ricerca qui presentata è che in primo luogo The Chymical Wedding si inserisce fra i romanzi della letteratura anglofona che attivano il codice alchemico. In particolare, Clarke ha utilizzato il simbolo delle nozze chimiche per definire i rapporti tra i personaggi, che lentamente inizieranno ad interagire tra di loro e istituiranno una relazione di mutua integrazione e collaborazione. Questo processo di integrazione e unione nella complementarietà avviene nel mondo interno di ciascun personaggio, nella dinamica conscio-inconscio, in quanto essi, nell’indagare il mistero delle nozze chimiche, intraprenderanno anche un percorso di crescita personale, la quale manifesta importanti momenti di avanzamento prima di tutto nella dimensione onirica. Infine, abbiamo interpretato l’evoluzione dei protagonisti come un processo di individuazione junghiano, che ha come scopo il diventare “un individuo completo” e liberare il S

Assessment of microbial cell viability in municipal sludge after microwave and ultrasound and subsequent impacts on anaerobic digestion

Anaerobic digestion (AD) is an effective method of waste sludge stabilization with lower costs and energy footprints; recently, researchers have begun to optimize the enhancement of AD by pretreating sludge prior to AD. In this study, two sludge pretreatment methods, ultrasonication (US) and microwave irradiation (MW) were compared to determine the relative amounts of microbial inactivation within pretreated secondary sludge, and how this cellular destruction translates to AD processes. The intensities chosen for MWpretreatment were based on previous studies using sludge from the wastewater treatment plant in Kelowna, BC, Canada. The intensity range chosen for MW possessed specific energy inputs of 2.17, 2.62, 4.89, and 6.48 kJ/g total solids (TS). For comparison purposes, US intensities were calculated to be 2.37, 4.74, 6.73, 23.09, and 27.71 kJ/g TS. Using a novel approach, the extent of cellular destruction caused at these intensities was measured using microbial viability fluorophore assays: 1) a molecular assay to measure live and dead cells and 2) a fluorescein diacetate assay to measure relative metabolic activity of sludge micro-organisms. From the results of the viability assays, it was determined that MW had the greatest effect on cells, having several times greater cell death and inactivation than both US-pretreated and non-pretreated sludge, even at lower specific energy values. Additionally, a MW and US intensity with similar specific energy inputs (2.62 kJ/gTS and 2.37 kJ/gTS, respectively) were applied to feed sludge of bench-scale digesters to compare effects on AD over three consecutive sludge retention times (SRTs), at 20 d, 14 d, and 7 d. The MW-fed digester had the highest overall methane production (248.2 L CH₄/kg volatile solids), greatest pathogen removal (73.4% and 69.8% less fecal coliforms over control during the 14 d and 7 d SRTs, respectively), and greatest solids removal (44.2% TS reduction). Interestingly, fecal coliform concentrations in the digester fed USpretreated sludge increased over the control for both the 14 d (31%) and 7 d (39.6%) SRT. All digesters possessed positive net energy production over the three chosen SRTs.Applied Science, Faculty ofEngineering, School of (Okanagan)Graduat

Cloning, functional characterization and genomic organization of 1,8-cineole synthases from Lavandula

Several members of the genus Lavandula produce valuable essential oils (EOs) that are primarily constituted of the low molecular weight isoprenoids, particularly monoterpenes. We isolated over 8,000 ESTs from the glandular trichomes of L. x intermedia flowers (where bulk of the EO is synthesized) to facilitate the discovery of genes that control the biosynthesis of EO constituents. The expression profile of these ESTs in L. x intermedia and its parents L. angustifolia and L. latifolia was established using microarrays. The resulting data highlighted a differentially expressed, previously uncharacterized cDNA with strong homology to known 1,8-cineole synthase (CINS) genes. The ORF, excluding the transit peptide, of this cDNA was expressed in E. coli, purified by Ni-NTA agarose affinity chromatography and functionally characterized in vitro. The ca. 63 kDa bacterially produced recombinant protein, designated L. x intermedia CINS (LiCINS), converted geranyl diphosphate (the linear monoterpene precursor) primarily to 1,8-cineole with K ( m ) and k ( cat ) values of 5.75 μM and 8.8 × 10-³ s-¹, respectively. The genomic DNA of CINS in the studied Lavandula species had identical exon-intron architecture and coding sequences, except for a single polymorphic nucleotide in the L. angustifolia ortholog which did not alter protein function. Additional nucleotide variations restricted to L. angustifolia introns were also observed, suggesting that LiCINS was most likely inherited from L. latifolia. The LiCINS mRNA levels paralleled the 1,8-cineole content in mature flowers of the three lavender species, and in developmental stages of L. x intermedia inflorescence indicating that the production of 1,8 cineole in Lavandula is most likely controlled through transcriptional regulation of LiCINS. [This is a post-peer-review, pre-copyedit version of an article published in Plant Molecular Biology. The final authenticated version is available online at: http://dx.doi.org/10.1007/s11103-012-9920-3]Arts and Sciences, Irving K. Barber School of (Okanagan)Biology, Department of (Okanagan)ReviewedFacult