The use of EORTC measures in daily clinical practice-A synopsis of a newly developed manual.

Cancer has increasingly become a chronic condition and the routine collection of patient-reported outcomes (PROs) like quality of life is widely recommended for clinical practice. Nonetheless, the successful implementation of PROs is still a major challenge, although common barriers to and facilitators of their beneficial use are well known. To support health care professionals and other stakeholders in the implementation of the EORTC PRO measures, the EORTC Quality of Life Group provides guidance on issues considered important for their use in daily clinical practice. Herein, we present an outline of the newly developed "'Manual for the use of EORTC measures in daily clinical practice", covering the following issues: * a rationale for using EORTC measures in routine care *selection of EORTC measures, timing of assessments, scoring and presentation of results * aspects of a strategic implementation * electronic data assessment and telemonitoring, and * further use of EORTC measures and ethical considerations. Next to an extensive overview of currently available literature, the manual specifically focuses on knowledge about EORTC measures to give evidence-based recommendations whenever possible and to encourage readers and end-users of EORTC measures to contribute to further needed high-quality research. The manual will be accessible on the EORTC Quality of Life Group website's homepage and will be periodically updated to take into account any new knowledge due to medical, technical, regulatory and scientific advances

Trajectories of health-related quality of life and symptom burden in patients with advanced cancer towards the end of life: Longitudinal results from the eQuiPe study

Background: Support for health-related quality of life (HRQOL) is an essential part of cancer care in the final stages of life, yet empirical guidance regarding HRQOL and symptom trajectories is lacking. Aim: To assess the change in HRQOL and symptom burden in the last year of life in patients with advanced cancer and its association with health care–related factors, cancer-specific treatment, and comorbidity. Methods: A prospective, multicenter, observational study in patients with advanced cancer (eQuiPe). Three monthly questionnaires included European Organization for Research and Treatment of Cancer Quality of Life-C30 and reported continuity of care. Multivariable mixed-effects analysis was used to assess the association between HRQOL and health care–related factors. Results: A total of 762 deceased patients were included with a mean age of 66 (SD, 10) years and 52% were male. The most common primary tumors were lung (29%), colorectal (20%), and breast cancer (13%). Mean overall HRQOL decreased in the last 9 months of life, with the greatest decrease in the last 3 months (β –16.2). Fatigue, pain, appetite loss, dyspnea, constipation, and nausea worsened significantly in the last year of life. Multimorbidity (β –7.5) and a better reported continuity of care (β 0.7) were both significantly associated with the trajectory of HRQOL. Conclusion: Mean overall HRQOL begins to decline 9 months before death, highlighting the need for early identification and (re)assessment of different symptoms as aspects of HRQOL follow different trajectories. Multimorbidity and reported continuity of care may be associated with the trajectory of HRQOL

Staff-resident interactions in long-term care for people with dementia: the role of meeting psychological needs in achieving residents' well-being.

noObjectives: The aim of this study is to explore the extent to which staff–resident interactions address or undermine residents’ psychological needs and how such interactions are associated with residents’ well-being. Method: Data on staff–resident interactions and residents’ well-being were collected for 51 residents from nine long-term care settings using dementia care mapping (DCM). DCM yields a count and detailed description of staff–resident interactions that either address (personal enhancers – PEs) or undermine (personal detractions – PDs) residents’ psychological needs, and every 5-minute scores for each resident's mood and engagement (ME-value). The relationship between PEs and PDs and well-being was analysed by studying residents’ ME-values before and three time frames after a PE or PD occurred. Results: A total of 76 PEs and 33 PDs were observed. The most common PEs were those addressing psychological needs for comfort and occupation. However residents’ well-being increased most often after PEs that addressed residents’ need for identity, attachment and inclusion. The most common PDs were those which undermined the need for comfort, inclusion and occupation. Residents’ well-being decreased most often after PDs that undermined the need for comfort. Conclusion: Increasing interactions which address residents’ need for attachment, identity and inclusion and eliminating interactions which undermine residents’ need for comfort may be particularly important in achieving residents’ well-being. In the long run, residents’ well-being could be achieved by staff availing of the opportunities to empower and facilitate residents, thus meeting their needs for occupation. These findings provide directions for training in person-centred care

The RNA template channel of the RNA-dependent RNA polymerase as a target for development of antiviral therapy of multiple genera within a virus family

The genus Enterovirus of the family Picornaviridae contains many important human pathogens (e.g., poliovirus, coxsackievirus, rhinovirus, and enterovirus 71) for which no antiviral drugs are available. The viral RNA-dependent RNA polymerase is an attractive target for antiviral therapy. Nucleoside-based inhibitors have broad-spectrum activity but often exhibit off-target effects. Most non-nucleoside inhibitors (NNIs) target surface cavities, which are structurally more flexible than the nucleotide-binding pocket, and hence have a more narrow spectrum of activity and are more prone to resistance development. Here, we report a novel NNI, GPC-N114 (2,2'-[(4-chloro-1,2-phenylene)bis(oxy)]bis(5-nitro-benzonitrile)) with broad-spectrum activity against enteroviruses and cardioviruses (another genus in the picornavirus family). Surprisingly, coxsackievirus B3 (CVB3) and poliovirus displayed a high genetic barrier to resistance against GPC-N114. By contrast, EMCV, a cardiovirus, rapidly acquired resistance due to mutations in 3Dpol. In vitro polymerase activity assays showed that GPC-N114 i) inhibited the elongation activity of recombinant CVB3 and EMCV 3Dpol, (ii) had reduced activity against EMCV 3Dpol with the resistance mutations, and (iii) was most efficient in inhibiting 3Dpol when added before the RNA template-primer duplex. Elucidation of a crystal structure of the inhibitor bound to CVB3 3Dpol confirmed the RNA-binding channel as the target for GPC-N114. Docking studies of the compound into the crystal structures of the compound-resistant EMCV 3Dpol mutants suggested that the resistant phenotype is due to subtle changes that interfere with the binding of GPC-N114 but not of the RNA template-primer. In conclusion, this study presents the first NNI that targets the RNA template channel of the picornavirus polymerase and identifies a new pocket that can be used for the design of broad-spectrum inhibitors. Moreover, this study provides important new insight into the plasticity of picornavirus polymerases at the template binding site.status: publishe

The RNA Template Channel of the RNA-Dependent RNA Polymerase as a Target for Development of Antiviral Therapy of Multiple Genera within a Virus Family

The genus Enterovirus of the family Picornaviridae contains many important human pathogens (e.g., poliovirus, coxsackievirus, rhinovirus, and enterovirus 71) for which no antiviral drugs are available. The viral RNA-dependent RNA polymerase is an attractive target for antiviral therapy. Nucleoside-based inhibitors have broad-spectrum activity but often exhibit off-target effects. Most non-nucleoside inhibitors (NNIs) target surface cavities, which are structurally more flexible than the nucleotide-binding pocket, and hence have a more narrow spectrum of activity and are more prone to resistance development. Here, we report a novel NNI, GPC-N114 (2,2'-[(4-chloro-1,2-phenylene)bis(oxy)]bis(5-nitro-benzonitrile)) with broad-spectrum activity against enteroviruses and cardioviruses (another genus in the picornavirus family). Surprisingly, coxsackievirus B3 (CVB3) and poliovirus displayed a high genetic barrier to resistance against GPC-N114. By contrast, EMCV, a cardiovirus, rapidly acquired resistance due to mutations in 3Dpol. In vitro polymerase activity assays showed that GPC-N114 i) inhibited the elongation activity of recombinant CVB3 and EMCV 3Dpol, (ii) had reduced activity against EMCV 3Dpol with the resistance mutations, and (iii) was most efficient in inhibiting 3Dpol when added before the RNA template-primer duplex. Elucidation of a crystal structure of the inhibitor bound to CVB3 3Dpol confirmed the RNA-binding channel as the target for GPC-N114. Docking studies of the compound into the crystal structures of the compound-resistant EMCV 3Dpol mutants suggested that the resistant phenotype is due to subtle changes that interfere with the binding of GPC-N114 but not of the RNA template-primer. In conclusion, this study presents the first NNI that targets the RNA template channel of the picornavirus polymerase and identifies a new pocket that can be used for the design of broad-spectrum inhibitors. Moreover, this study provides important new insight into the plasticity of picornavirus polymerases at the template binding site

Characterization of the inhibitory mechanism of GPC-N114 on CVB3 3D^pol.

<p>(A) GPC-N114 does not compete with UTP. The experiment was performed for CVB3 3D^pol as described in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004733#ppat.1004733.g002" target="_blank">Fig. 2E</a> for EMCV 3D^pol. (B) GPC-N114 is more efficient when added to the reaction mix before the template-primer. Order-of-addition experiments were performed by measuring CVB3 3D^pol elongation activity when GPC-N114 was added to the reaction mix before or after poly(rA)/dT15. A typical of three independent experiments is presented.</p

The binding site of GPC-N114 on CVB3 3D^pol.

<p>(A) Ribbon diagram of the CVB3 3D^pol structure (green). The inhibitor bound to the bottom of the template channel is shown in stick representation in yellow. (B) Interaction network between GPC-N114 and its binding pocket of CVB3 3D^pol represented by surfaces. The polymerase residues in direct contact with the inhibitor are shown with carbon atoms in green and explicitly labeled. Hydrogen bonds are depicted as dashed lines. (C) Top down cross-sectional view of the molecular surface of CVB3 3D^pol with the electrostatic potential, represented in blue and red for positive and negative charges, respectively. The trajectory of the RNA template (green) is compared with the position of the GPC-N114 inhibitor (yellow). The inset box shows a close up of the position of the inhibitor that is coincident with that of the template acceptor nucleotide. The predicted position of the RNA template was determined using the FMDV 3D^pol–RNA complex (PDB entry 1WNE) as a guide.</p

GPC-N114 inhibits picornavirus replication.

<p>(A) Structural formula of GPC-N114. (B) Representative dose-response curves of multicycle CPE-reduction assays for CVB3, PV1, and EV71. CPE was quantified by MTS assay at 3 d p.i. and is expressed as percentage of uninfected, untreated controls. (C, D) Antiviral activity of GPC-N114 against CVB3 and EMCV. BGM cells were infected with CVB3 (left panels) or EMCV (right panels) at an MOI of 0.1. Immediately after infection, GPC-N114 was added at the indicated concentrations (C) or at 10 μM (D). The enterovirus inhibitor guanidine hydrochloride (GuHCl) and the cardiovirus inhibitor dipyridamole were included as controls. Virus titers were determined by endpoint titration after 8 h (C) or at the indicated times p.i. (D). Experiments were performed in triplicate and mean values ± SD are depicted. (E) Antiviral activity of GPC-N114 against a range of picornaviruses. Cells were infected with the indicated viruses at an MOI of 0.5 after which 10 μM GPC-N114 was added. Virus titers were determined at 8 h p.i. Experiments were performed in triplicate and mean values ± SD are depicted. (F) GPC-N114 inhibits viral RNA replication. RNA of subgenomic replicons of CVB3 or EMCV was transfected into BGM cells. Subsequently, cells were treated either with 0.1% DMSO, 10 μM GPC-N114, 2 mM GuHCl, or 80 μM dipyridamole. Firefly luciferase levels were determined 2, 4, 6, and 8 h after RNA transfection to assess the level of replication and translation. Experiments were performed in triplicate and mean values ± SD are depicted.</p

Data collection and refinement statistics.

<p>† Rwork = ∑hkl ||Fobs(hkl)|—|Fcalc(hkl)|| / ∑hkl |Fobs(hkl)|, where Fobs and Fcalc are the structure factors, deduced from measured intensities and calculated from the model, respectively.</p><p>‡ Rfree = as for Rwork but for 5% of the total reflections chosen at random and omitted from refinement.</p><p>Data collection and refinement statistics.</p

Effects of mutations M300V and I304V on the GPC-N114 binding site in EMCV 3D^pol.

<p>(A) Surface representation of the putative GPC-N114 binding pocket in EMCV 3D^pol (slate blue) with the modeled inhibitor depicted in atom color (carbon atoms in yellow). (B-D) Environment of 300 and 304 residues on EMCV wt and mutants 3D^pol. Motifs A and C are represented in transparent grey and motif B in solid color: (B) grey (wt), (C) cyan (M300V) and (D) red (I304V). Residues interacting with motif B α-helix are shown as sticks colored by atom. The GPC-N114 binding site on CVB3 3D^pol is represented as a yellow surface.</p