Clinical Activity and Quality of Life Indices Are Valid Across Ulcerative Colitis But Not Crohn’s Disease Phenotypes

Background Clinical activity and quality of life (QOL) indices assess disease activity in Crohn’s disease (CD) and ulcerative colitis (UC). However, a paucity of data exists on the validity of these indices according to disease characteristics. Aims To examine the correlation between QOL and clinical activity indices and endoscopic disease activity according to disease characteristics. Methods We used a prospective registry to identify CD and UC patients ≥18 years old with available information on Short Inflammatory Bowel Disease Questionnaire scores (SIBDQ), Harvey–Bradshaw Index (HBI) and simple endoscopic scores for CD (SES-CD), and Simple Clinical Colitis Activity Index (SCCAI) and Mayo endoscopic score for UC. We used Spearman rank correlations to calculate correlations between indices and Fisher transformation to compare correlations across disease characteristics. Results Among 282 CD patients, we observed poor correlation between clinical activity and QOL indices to SES-CD with no differences in correlation according to disease characteristics. Conversely, among 226 UC patients, clinical activity and QOL had good correlation to Mayo endoscopic score (r = 0.55 and −0.56, respectively) with better correlations observed with left-sided versus extensive colitis (r = 0.73 vs. 0.45, p = 0.005) and shorter duration of disease (r = 0.61 vs. 0.37, p = 0.04). Conclusions Our data suggest good correlation between SCCAI and endoscopic disease activity in UC, particularly in left-sided disease. Poor correlations between HBI or SIBDQ and SES-CD appear to be consistent across different disease phenotypes.American Gastroenterological Associatio

Graphene plasmonics: A platform for strong light-matter interaction

Graphene plasmons provide a suitable alternative to noble-metal plasmons because they exhibit much larger confinement and relatively long propagation distances, with the advantage of being highly tunable via electrostatic gating. We report strong light- matter interaction assisted by graphene plasmons, and in particular, we predict unprecedented high decay rates of quantum emitters in the proximity of a carbon sheet, large vacuum Rabi splitting and Purcell factors, and extinction cross sections exceeding the geometrical area in graphene ribbons and nanometer-sized disks. Our results provide the basis for the emerging and potentially far-reaching field of graphene plasmonics, offering an ideal platform for cavity quantum electrodynamics and supporting the possibility of single-molecule, single-plasmon devices.Comment: 39 pages, 15 figure

No evidence for higher rates of hepatocellular carcinoma after direct-acting antiviral treatment: a meta-analysis

Aim: Hepatitis C virus (HCV) is the leading cause of hepatocellular carcinoma (HCC) in the United States. Achieving sustained viral response with interferon (IFN) treatment reduces the risk from 3%-5% to 0.5%-1% annually. Several studies reported unexpectedly high rates of HCC after treatment with direct-acting antivirals (DAAs). The aim of our study was to compare HCC rates in DAA-, IFN-treated and untreated populations.Methods: A literature search was conducted using ScienceDirect, Ovid®, Web of Science and MEDLINE through January 2019. Studies were included if they measured rates of de novo or recurrent HCC (following curative treatment) in HCV-infected persons. We included 138 studies (n = 177,512). Simple pooling of data and meta-analysis were performed, using the random effects method.Results: Mean age was higher in the DAA-treated vs. IFN-treated group (58.4 years vs. 52.6 years; P = 0.0073), as were diabetes prevalence (34.5% vs. 11.7%; P ≤ 0.001) and incident cirrhosis (47.8% vs. 34.2%, P = 0.0017). The incidence rate of de novo HCC was 2.01/100 person-years (py) (95%CI: 1.38, 2.67) in the DAA group and 1.45/100py (95%CI: 0.98, 1.94) in the IFN-treated group. HCC recurred at 16.76/100py (95%CI: 10.75, 22.91) in the DAA-treated group vs. 20.04/100py (95%CI: 2.58, 45.21) after IFN. After adjusting for factors such as age and cirrhosis, the hazard ratio was 0.58 (95%CI: 0.20, 1.07) for HCC occurrence and 0.59 (95%CI: 0.24, 1.03) for HCC recurrence after DAA treatment compared to IFN-based treatment.Conclusion: We did not find evidence for increased rates of HCC in DAA-treated compared with IFN-treated patients. Compared to those treated with IFN, older patients with additional risk factors for HCC were treated with DAAs. This imbalance appears to explain the higher numerical incidence of HCC among DAA-treated patients

A cell system for phenotypic screening of modifiers of SMN2 gene expression and function.

Spinal muscular atrophy (SMA) is an inherited neurodegenerative disease caused by homozygous inactivation of the SMN1 gene and reduced levels of the survival motor neuron (SMN) protein. Since higher copy numbers of the nearly identical SMN2 gene reduce disease severity, to date most efforts to develop a therapy for SMA have focused on enhancing SMN expression. Identification of alternative therapeutic approaches has partly been hindered by limited knowledge of potential targets and the lack of cell-based screening assays that serve as readouts of SMN function. Here, we established a cell system in which proliferation of cultured mouse fibroblasts is dependent on functional SMN produced from the SMN2 gene. To do so, we introduced the entire human SMN2 gene into NIH3T3 cell lines in which regulated knockdown of endogenous mouse Smn severely decreases cell proliferation. We found that low SMN2 copy number has modest effects on the cell proliferation phenotype induced by Smn depletion, while high SMN2 copy number is strongly protective. Additionally, cell proliferation correlates with the level of SMN activity in small nuclear ribonucleoprotein assembly. Following miniaturization into a high-throughput format, our cell-based phenotypic assay accurately measures the beneficial effects of both pharmacological and genetic treatments leading to SMN upregulation. This cell model provides a novel platform for phenotypic screening of modifiers of SMN2 gene expression and function that act through multiple mechanisms, and a powerful new tool for studies of SMN biology and SMA therapeutic development

Genetic modulation of SMN-dependent proliferation in NIH3T3 cells.

<p>Dose-response analysis of lentiviral-mediated human SMN expression on cell proliferation of SMN-deficient NIH3T3-Smn_RNAi and NIH3T3-SMN2_low/Smn_RNAi cells. In these experiments, NIH3T3 cells were cultured for 5 days with Dox prior to seeding in six replicate wells of a 96-well plate in the presence of Dox. Vehicle or increasing amounts of SMN-expressing lentivirus were added 4 hours later. For each group and treatment, cell number was determined at 5 days post-plating and normalized to that of the corresponding vehicle-treated cells. Data are represented as mean and SEM (n = 6; * = p<0.05; ** = p<0.01; *** = p<0.001; one-way ANOVA).</p

Development of <i>SMN2</i>-containing NIH3T3 cell lines with inducible knockdown of endogenous Smn.

<p>(A) Schematic representation of the exon-intron structure of the human <i>SMN2</i> gene used to establish NIH3T3-SMN2/Smn_RNAi cell lines (introns not drawn to scale). (B) Analysis of <i>SMN2</i> gene copy number in the indicated NIH3T3 cell lines by qPCR. Data were normalized to the <i>Gapdh</i> gene and expressed relative to the NIH3T3-SMN2_low/Smn_RNAi cell line. Data are represented as mean and SEM (n = 3; ** = p<0.01; t-test). (C) Analysis of total SMN2 mRNA levels in the indicated NIH3T3 cell lines by RT-qPCR. Data were normalized to Gapdh mRNA and expressed relative to the NIH3T3-SMN2_low/Smn_RNAi cell line. Data are represented as mean and SEM (n = 3; *** = p<0.001, t-test). (D) Analysis of SMN2 exon 7 splicing in the indicated NIH3T3 cell lines by radioactive RT-PCR. The levels of exon 7 inclusion are shown at the bottom.</p

Subcellular localization of the human SMN protein in NIH3T3-SMN2/Smn_RNAi cell lines.

<p>Indirect immunofluorescence and confocal microscopy analysis of NIH3T3-Smn_RNAi (A and D), NIH3T3-SMN2_low/Smn_RNAi (B and E) and NIH3T3-SMN2_high/Smn_RNAi (C and F) cell lines using monoclonal antibodies specific to human SMN (hSMN, panels A–C) or both mouse and human SMN (SMN, panels D–F). Scale bar, 10 µm.</p

Pharmacological modulation of SMN-dependent proliferation in NIH3T3 cells.

<p>(A) Dose-response analysis of VPA treatment on cell proliferation of SMN-deficient NIH3T3-Smn_RNAi and NIH3T3-SMN2_low/Smn_RNAi cells. In these experiments, NIH3T3 cells were cultured for 5 days with Dox prior to seeding in six replicate wells of a 96-well plate in the presence of Dox. Vehicle or increasing amounts of VPA were added 4 hours later. For each group and treatment, cell number was determined at 5 days post-plating and normalized to that of the corresponding vehicle-treated cells. Data are represented as mean and SEM (n = 6; * = p<0.05; ** = p<0.01; *** = p<0.001; one-way ANOVA). (B–C) RT-qPCR analysis of mouse Smn and human SMN2 mRNA levels in NIH3T3-Smn_RNAi (B) and NIH3T3-SMN2_low/Smn_RNAi (C) cells cultured with Dox for 3 days and then in the presence of either water (Vehicle) or 1 mM VPA for additional 4 days. Data from triplicate RT-qPCR experiments normalized to Gapdh mRNA are represented as mean and SEM (** = p<0.01; t-test). (D–E) Western blot analysis of SMN protein levels in NIH3T3-Smn_RNAi (D) and NIH3T3-SMN2_low/Smn_RNAi (E) cells cultured with Dox for 3 days and then in the presence of water (Vehicle) or 1 mM VPA for additional 4 days. Blots were probed with an antibody that recognizes both mouse and human SMN proteins. SMN levels in VPA-treated relative to vehicle-treated cells are shown at the bottom.</p

Effect of human SMN2 expression on proliferation and snRNP assembly in Smn-deficient NIH3T3 cell lines.

<p>(A) Analysis of cell proliferation in wild-type (Control), NIH3T3-Smn_RNAi, NIH3T3-SMN2_low/Smn_RNAi and NIH3T3-SMN2_high/Smn_RNAi cell lines cultured with and without doxycycline for 7 days. For each cell line, the cell number ratio of Dox-treated cells versus untreated cells is expressed relative to that of wild-type cells, which is set to 1. Data are represented as mean and SEM (n≥3; *** = p<0.001; one-way ANOVA). (B) Analysis of U1 snRNP assembly in NIH3T3 cell lines. <i>In vitro</i> snRNP assembly experiments were carried out with radioactive U1 snRNA and extracts from wild-type NIH3T3 (Control), NIH3T3-Smn_RNAi, NIH3T3-SMN2_low/Smn_RNAi and SMN2_high/Smn_RNAi cells cultured with and without doxycycline for 7 days. For each cell line, the amounts of immunoprecipitated U1 snRNA were quantified and the RNA ratio in Dox-treated cells versus untreated cells is expressed relative to that of wild-type cells, which is set to 1. Data are represented as mean and SEM (n≥3; * = p<0.05; ** = p<0.01; *** = p<0.001; one-way ANOVA).</p