bluuebunny/crossref_metadata_2025_split huggingface dataset:
RelA Mutant
Enterococcus faecium
with Multiantibiotic Tolerance Arising in an Immunocompromised Host
Authors: Honsa Erin S. , Cooper Vaughn S. , Mhaissen Mohammed N. , Frank Matthew , Shaker Jessica , Iverson Amy , Rubnitz Jeffrey , Hayden Randall T. , Lee Richard E. , Rock Charles O. , Tuomanen Elaine I. , Wolf Joshua , Rosch Jason W. 
Published: January 2017

ABSTRACT

Serious bacterial infections in immunocompromised patients require highly effective antibacterial therapy for cure, and thus, this setting may reveal novel mechanisms by which bacteria circumvent antibiotics in the absence of immune pressure. Here, an infant with leukemia developed vancomycin-resistant
Enterococcus faecium
(VRE) bacteremia that persisted for 26 days despite appropriate antibiotic therapy. Sequencing of 22 consecutive VRE isolates identified the emergence of a single missense mutation (L152F) in
relA
, which constitutively activated the stringent response, resulting in elevated baseline levels of the alarmone guanosine tetraphosphate (ppGpp). Although the mutant remained susceptible to both linezolid and daptomycin in clinical MIC testing and during planktonic growth, it demonstrated tolerance to high doses of both antibiotics when growing in a biofilm. This biofilm-specific gain in resistance was reflected in the broad shift in transcript levels caused by the mutation. Only an experimental biofilm-targeting ClpP-activating antibiotic was able to kill the mutant strain in an established biofilm. The
relA
mutation was associated with a fitness trade-off, forming smaller and less-well-populated biofilms on biological surfaces. We conclude that clinically relevant
relA
mutations can emerge during prolonged VRE infection, causing baseline activation of the stringent response, subsequent antibiotic tolerance, and delayed eradication in an immunocompromised state.

IMPORTANCE
The increasing prevalence of antibiotic-resistant bacterial pathogens is a major challenge currently facing the medical community. Such pathogens are of particular importance in immunocompromised patients as these individuals may favor emergence of novel resistance determinants due to lack of innate immune defenses and intensive antibiotic exposure. During the course of chemotherapy, a patient developed prolonged bacteremia with vancomycin-resistant
Enterococcus faecium
that failed to clear despite multiple front-line antibiotics. The consecutive bloodstream isolates were sequenced, and a single missense mutation identified in the
relA
gene, the mediator of the stringent response. Strains harboring the mutation had elevated baseline levels of the alarmone and displayed heightened resistance to the bactericidal activity of multiple antibiotics, particularly in a biofilm. Using a new class of compounds that modulate ClpP activity, the biofilms were successfully eradicated. These data represent the first clinical emergence of mutations in the stringent response in vancomycin-resistant entereococci.
FOSL1 Inhibits Type I Interferon Responses to Malaria and Viral Infections by Blocking TBK1 and TRAF3/TRIF Interactions
Authors: Cai Baowei , Wu Jian , Yu Xiao , Su Xin-zhuan , Wang Rong-Fu 
Published: January 2017

ABSTRACT
Innate immune response plays a critical role in controlling invading pathogens, but such an immune response must be tightly regulated. Insufficient or overactivated immune responses may lead to harmful or even fatal consequences. To dissect the complex host-parasite interactions and the molecular mechanisms underlying innate immune responses to infections, here we investigate the role of FOS-like antigen 1 (FOSL1) in regulating the host type I interferon (IFN-I) response to malaria parasite and viral infections. FOSL1 is known as a component of a transcription factor but was recently implicated in regulating the IFN-I response to malaria parasite infection. Here we show that FOSL1 can act as a negative regulator of IFN-I signaling. Upon stimulation with poly(I:C), malaria parasite-infected red blood cells (iRBCs), or vesicular stomatitis virus (VSV), FOSL1 “translocated” from the nucleus to the cytoplasm, where it inhibited the interactions between TNF receptor-associated factor 3 (TRAF3), TIR domain-containing adapter inducing IFN-β (TRIF), and Tank-binding kinase 1 (TBK1) via impairing K63-linked polyubiquitination of TRAF3 and TRIF. Importantly, FOSL1 knockout chimeric mice had lower levels of malaria parasitemia or VSV titers in peripheral blood and decreased mortality compared with wild-type (WT) mice. Thus, our findings have identified a new role for FOSL1 in negatively regulating the host IFN-I response to malaria and viral infections and have identified a potential drug target for controlling malaria and other diseases.

IMPORTANCE
Infections of pathogens can trigger vigorous host immune responses, including activation and production of type I interferon (IFN-I). In this study, we investigated the role of FOSL1, a molecule previously known as a transcription factor, in negatively regulating IFN-I responses to malaria and viral infections. We showed that FOSL1 was upregulated and translocated into the cytoplasm of cells after stimulation for IFN-I production. FOSL1 could affect TRAF3 and TRIF ubiquitination and consequently impaired the association of TRAF3, TRIF, and TBK1, leading to inhibition of IFN-I signaling.
In vivo
experiments with FOSL1 knockout chimeric mice further validated the negative role of FOSL1 in IFN-I production and antimicrobial responses. This report reveals a new functional role for FOSL1 in IFN-I signaling and dissects the mechanism by which FOSL1 regulates IFN-I responses to malaria and viral infections, which can be explored as a potential drug target for disease control and management.
Modulation of microRNA-mRNA Target Pairs by Human Papillomavirus 16 Oncoproteins
Authors: Harden Mallory E. , Prasad Nripesh , Griffiths Anthony , Munger Karl 
Published: January 2017

ABSTRACT
The E6 and E7 proteins are the major oncogenic drivers encoded by high-risk human papillomaviruses (HPVs). While many aspects of the transforming activities of these proteins have been extensively studied, there are fewer studies that have investigated how HPV E6/E7 expression affects the expression of cellular noncoding RNAs. The goal of our study was to investigate HPV16 E6/E7 modulation of cellular microRNA (miR) levels and to determine the potential consequences for cellular gene expression. We performed deep sequencing of small and large cellular RNAs in primary undifferentiated cultures of human foreskin keratinocytes (HFKs) with stable expression of HPV16 E6/E7 or a control vector. After integration of the two data sets, we identified 51 differentially expressed cellular miRs associated with the modulation of 1,456 potential target mRNAs in HPV16 E6/E7-expressing HFKs. We discovered that the degree of differential miR expression in HFKs expressing HPV16 E6/E7 was not necessarily predictive of the number of corresponding mRNA targets or the potential impact on gene expression. Additional analyses of the identified miR-mRNA pairs suggest modulation of specific biological activities and biochemical pathways. Overall, our study supports the model that perturbation of cellular miR expression by HPV16 E6/E7 importantly contributes to the rewiring of cellular regulatory circuits by the high-risk HPV E6 and E7 proteins that contribute to oncogenic transformation.

IMPORTANCE
High-risk human papillomaviruses (HPVs) are the causative agents of almost all cervical cancers and many other cancers, including anal, vaginal, vulvar, penile, and oropharyngeal cancers. Despite the availability of efficacious HPV vaccines, it is critical to determine how HPVs cause cancer, as many people remain unvaccinated and the vaccine does not prevent cancer development in individuals who are already infected. Two HPV proteins, E6 and E7, are the major drivers of cancer development, and much remains to be learned about how the expression of these viral proteins reprograms infected cells, ultimately resulting in cancer development. Small, noncoding human RNAs, termed microRNAs (miRs), regulate gene expression and have been implicated in almost all human cancers, including HPV-associated cancers. Our study provides a comprehensive analysis of how E6 and E7 alter the expression of human miRs and how this potentially impacts cellular gene expression, which may contribute to cancer development.
Healthcare use and expenditure for diabetes in Bangladesh
Authors: Shariful Islam Sheikh Mohammed , Lechner Andreas , Ferrari Uta , Laxy Michael , Seissler Jochen , Brown Jonathan , Niessen Louis W , Holle Rolf 
Published: January 2017

BackgroundDiabetes imposes a huge social and economic impact on nations. However, information on the costs of treating and managing diabetes in developing countries is limited. The aim of this study was to estimate healthcare use and expenditure for diabetes in Bangladesh.MethodsWe conducted a matched case–control study between January and July 2014 among 591 adults with diagnosed diabetes mellitus (DMs) and 591 age-matched, sex-matched and residence-matched persons without diabetes mellitus (non-DMs). We recruited DMs from consecutive patients and non-DMs from accompanying persons in the Bangladesh Institute of Health Science (BIHS) hospital in Dhaka, Bangladesh. We estimated the impact of diabetes on healthcare use and expenditure by calculating ratios and differences between DMs and non-DMs for all expenses related to healthcare use and tested for statistical difference using Student's t-tests.ResultsDMs had two times more days of inpatient treatment, 1.3 times more outpatient visits, and 9.7 times more medications than non-DMs (all p<0.005). The total annual per capita expenditure on medical care was 6.1 times higher for DMs than non-DMs (US$635 vs US$104, respectively). Among DMs, 9.8% reported not taking any antidiabetic medications, 46.4% took metformin, 38.7% sulfonylurea, 40.8% insulin, 38.7% any antihypertensive medication, and 14.2% took anti-lipids over the preceding 3 months.ConclusionsDiabetes significantly increases healthcare use and expenditure and is likely to impose a huge economic burden on the healthcare systems in Bangladesh. The study highlights the importance of prevention and optimum management of diabetes in Bangladesh and other developing countries, to gain a strong economic incentive through implementing multisectoral approach and cost-effective prevention strategies.
System-wide analysis of health financing equity in Cambodia: a study protocol
Authors: Wiseman Virginia , Asante Augustine , Ir Por , Limwattananon Supon , Jacobs Bart , Liverani Marco , Hayen Andrew , Jan Stephen 
Published: January 2017

BackgroundTo assess progress towards universal health coverage, countries like Cambodia require evidence on equity in the financing and distribution of healthcare benefits. This evidence must be based on a system-wide perspective that recognises the complex roles played by the public and private sectors in many contemporary healthcare systems.ObjectiveTo undertake a system-wide assessment of who pays and who benefits from healthcare in Cambodia and to understand the factors influencing this.MethodsFinancing and benefit incidence analysis will be used to calculate the financing burden and distribution of healthcare benefits across socioeconomic groups. Data on healthcare usage, living standards and self-assessed health status will be derived from a cross-sectional household survey designed for this study involving a random sample of 5000 households. This will be supplemented by secondary data from the Cambodian National Health Accounts 2014 and the Cambodian Socioeconomic Survey (CSES) 2014. We will also collect qualitative data through focus group discussions and in-depth interviews to inform the interpretation of the quantitative analyses.Potential impactThis study will produce previously unavailable information on who pays for, and who benefits from, health services across the entire health system of Cambodia. This evidence comes at a critical juncture in healthcare reform in South-East Asia with so many countries seeking guidance on the equity impact of their current financing arrangements that include a complex mix of public and private providers.
Association of erythrocyte methotrexate-polyglutamate levels with the efficacy and hepatotoxicity of methotrexate in patients with rheumatoid arthritis: a 76-week prospective study
Authors: Takahashi Chihiro , Kaneko Yuko , Okano Yutaka , Taguchi Hiroaki , Oshima Hisaji , Izumi Keisuke , Yamaoka Kunihiro , Takeuchi Tsutomu 
Published: January 2017

ObjectiveTo assess the utility of erythrocyte methotrexate-polyglutamate (MTX-PG) concentrations in determining the safety and efficacy of MTX in patients with rheumatoid arthritis (RA).Methods79 MTX-naïve patients with RA were enrolled in this prospective 76-week cohort study. MTX was initiated, and a predefined dose-escalation protocol was followed. Erythrocyte MTX-PG concentrations were measured using liquid chromatography. The associations of MTX-PG concentrations with disease activity and adverse events were analysed.ResultsDose escalation of MTX resulted in increased MTX-PG concentrations and a decrease in the mean Disease Activity Score in 28 joints (DAS28). A significant association was observed between total MTX-PG concentrations and ΔDAS28 at week 12 (β=−0.013, p=0.003) and at week 24 (β=−0.014, p=0.003). The maximum MTX-PG levels were significantly higher in patients presenting with elevated transaminases (≥100 IU/L) than in those without (146 vs 106 nmol/L, p=0.009). Receiver operating characteristic curve analysis revealed that a total MTX-PG concentrations of 83 nmol/L at week 12 was the threshold for a DAS28 improvement of ≥1.2 at week 24, and 105 nmol/L was the threshold for transaminases of ≥50 IU/L and 131 nmol/L for transaminases of ≥100 IU/L. MTX-PG concentrations were strongly influenced by body mass index and a serum albumin level.ConclusionsMTX-PG concentrations are a useful biomarker in MTX therapy, in terms of efficacy and safety.
Systematic review and network meta-analysis of the efficacy and safety of tumour necrosis factor inhibitor–methotrexate combination therapy versus triple therapy in rheumatoid arthritis
Authors: Fleischmann Roy , Tongbram Vanita , van Vollenhoven Ronald , Tang Derek H , Chung James , Collier David , Urs Shilpa , Ndirangu Kerigo , Wells George , Pope Janet 
Published: January 2017

ObjectiveClinical trials have not consistently demonstrated differences between tumour necrosis factor inhibitor (TNFi) plus methotrexate and triple therapy (methotrexate plus hydroxychloroquine plus sulfasalazine) in rheumatoid arthritis (RA). The study objective was to estimate the efficacy, radiographic benefits, safety and patient-reported outcomes of TNFi–methotrexate versus triple therapy in patients with RA.MethodsA systematic review and network meta-analysis (NMA) of randomised controlled trials of TNFi–methotrexate or triple therapy as one of the treatment arms in patients with an inadequate response to or who were naive to methotrexate was conducted. American College of Rheumatology 70% response criteria (ACR70) at 6 months was the prespecified primary endpoint to evaluate depth of response. Data from direct and indirect comparisons between TNFi–methotrexate and triple therapy were pooled and quantitatively analysed using fixed-effects and random-effects Bayesian models.ResultsWe analysed 33 studies in patients with inadequate response to methotrexate and 19 in patients naive to methotrexate. In inadequate responders, triple therapy was associated with lower odds of achieving ACR70 at 6 months compared with TNFi–methotrexate (OR 0.35, 95% credible interval (CrI) 0.19 to 0.64). Most secondary endpoints tended to favour TNFi–methotrexate in terms of OR direction; however, no clear increased likelihood of achieving these endpoints was observed for either therapy. The odds of infection were lower with triple therapy than with TNFi−methotrexate (OR 0.08, 95% CrI 0.00 to 0.57). There were no differences observed between the two regimens in patients naive to methotrexate.ConclusionsIn this NMA, triple therapy was associated with 65% lower odds of achieving ACR70 at 6 months compared with TNFi–methotrexate in patients with inadequate response to methotrexate. Although secondary endpoints numerically favoured TNFi–methotrexate, no clear differences were observed. The odds of infection were greater with TNFi–methotrexate. No differences were observed for patients naive to methotrexate. These results may help inform care of patients who fail methotrexate first-line therapy.
Abstract A01: Centrosome amplification induces autophagy and sensitizes to chloroquine
Authors: Denu Ryan A. , Burkard Mark E. 
Published: January 2017

Abstract
Autophagy is a catabolic process whereby autophagosomes engulf damaged organelles and proteins and deliver these contents to the lysosome for degradation and subsequent recycling. Proper autophagosome fusion with lysosomes depends on movement along intact microtubules, which are organized by centrosomes in human cells. We have discovered that autophagy is activated by centrosome amplification (CA), a numerical increase in centrosome number. CA is common in human cancers, particularly high-risk cancers, and higher autophagy is predicted to sensitize cancer cells with CA to autophagy inhibition. To test this hypothesis, we first evaluated autophagy in 341 human breast cancers with known CA status by quantifying p62/SQSTM1. Cancers with CA demonstrated a decrease in p62 expression, consistent with elevated autophagy. Next, we evaluated autophagy using western blot and immunofluorescence in two model cell lines of CA: doxycycline-inducible expression of polo-like kinase 4 (PLK4), an enzyme required for centrosome duplication. Cells with CA demonstrated elevated levels of autophagy by LC3 and p62 immunoblotting and immunofluorescence. Third, we tested autophagy inhibitors in our cell models of CA. Our results show that cells with CA are sensitized to the autophagy inhibitor chloroquine. Taken together, our results suggest that autophagy is upregulated by CA. We hypothesize that CA induces autophagy by increasing the number of lysosomal hubs (lysosomes tend to cluster around centrosomes) and facilitating transport to these hubs. These findings suggest CA as a biomarker for autophagy inhibition in high-risk cancers.
Citation Format: Ryan A. Denu, Mark E. Burkard. Centrosome amplification induces autophagy and sensitizes to chloroquine. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Targeting the Vulnerabilities of Cancer; May 16-19, 2016; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(1_Suppl):Abstract nr A01.
Abstract A23: Novel approach for targeting hepatocellular carcinoma cell survival: OSU-2S/Sorafenib combination
Authors: Omar Hany A. , Tolba Mai F. , Hung Jui-Hsiang , Al-Tel Taleb H. 
Published: January 2017

Abstract
Background: OSU-2S is a novel non-immunosuppressant anticancer agent that was designed and developed using fingolimod (Gilenya®) as a scaffold. OSU-2S was reported to have a substantial anticancer activity against hepatocellular carcinoma (HCC) through different mechanisms. The effectiveness of HCC treatment is often poor due to acquired resistance during the course of therapy. The current work was aimed to test the possible therapeutic synergy between OSU-2S and sorafenib, the multi-kinase inhibitor which is used as standard first-line systemic treatment for advanced HCC. Mechanistically, such combination can achieve optimal anticancer activity through synergistic targeting of the cancer cell survival signaling pathways.
Methods: The synergistic combination between OSU-2S and sorafenib was tested on three different HCC cell lines (Hep3B, Huh-7, HepG2) using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium) assay, colony formation, invasion, migration, immunofluorescence, and Western blot analysis.
Results: The combination incidences for OSU-2S/ Sorafenib obtained from MTT and colony formation assays were in the range of 0.4-0.8 at different dose levels which indicated synergistic anticancer activity. This synergy was confirmed by enhanced ability to activate apoptosis and to hinder several cell survival signaling pathways such as mitogen-activated protein kinase (MAPK) pathway. The synergy was mediated through the activation of the pro-apoptotic kinase, PKCδ and the augmentation of reactive oxygen species generation.
Conclusion: OSU-2S/ Sorafenib combination represents a novel therapeutically relevant approach to be introduced in HCC chemotherapeutic protocols which should effectively target cancer cells and improve the survival of HCC patients.
Citation Format: Hany A. Omar, Mai F. Tolba, Jui-Hsiang Hung, Taleb H. Al-Tel. Novel approach for targeting hepatocellular carcinoma cell survival: OSU-2S/Sorafenib combination. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Targeting the Vulnerabilities of Cancer; May 16-19, 2016; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(1_Suppl):Abstract nr A23.
Abstract A31: 14-3-3 binding sites in full-length and splice variant forms of BRAF V600E modulate the response to RAF inhibitors
Authors: Vido Michael J. , Le Kaitlyn , Rock Justin , Hartsough Edward J. , Dadpey Neda , Aplin Andrew E. 
Published: January 2017

Abstract
The serine/threonine kinase BRAF is mutated in 7% of human cancers, the most frequent alteration being a valine to glutamic acid substitution at amino acid 600 (V600E). BRAF V600E melanomas are highly sensitive to RAF inhibitors but the expression of aberrantly spliced BRAF V600E (BRAF V600E ΔEx) isoforms are associated with resistance in 13-30% of progressing patients. Compared to full-length BRAF V600E, BRAF V600E ΔEx exhibit enhanced dimerization and signaling via ERK1/2 during RAF inhibitor therapy; however, much remains unknown regarding their mechanism of action. The 14-3-3 protein binding sites, serine 365 (S365) and serine 729 (S729), play a complex role in regulating the activity and dimerization of RAF isoforms. All reported BRAF V600E ΔEx isoforms lose the N-terminal 14-3-3 binding site (S365) but retain the C-terminal site (S729); therefore, we analyzed the involvement of 14-3-3 binding sites in response to RAF inhibitor therapy. In full-length BRAF V600E, mutation of S365 to alanine promotes elevated MEK-ERK1/2 signaling and growth in the presence of RAF inhibitor. BRAF V600E ΔEx exhibits increased phosphorylation on S729 during RAF inhibitor therapy. Mutation of S729 to alanine renders the BRAF V600E ΔEx isoforms sensitive to RAF inhibitor, measured by reduced ERK1/2 phosphorylation and cell growth in vitro and in vivo. These data highlight the importance of BRAF V600E 14-3-3 binding sites in mediating targeted therapy resistance in mutant BRAF-driven melanoma.
Citation Format: Michael J. Vido, Kaitlyn Le, Justin Rock, Edward J. Hartsough, Neda Dadpey, Andrew E. Aplin. 14-3-3 binding sites in full-length and splice variant forms of BRAF V600E modulate the response to RAF inhibitors. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Targeting the Vulnerabilities of Cancer; May 16-19, 2016; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(1_Suppl):Abstract nr A31.