bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization of a Replication-Incompetent Pseudorabies Virus Mutant Lacking the Sole Immediate Early Gene IE180
Authors: Wu Brendan W. , Engel Esteban A. , Enquist Lynn W. 
Published: November 2014

ABSTRACT
The alphaherpesvirus pseudorabies virus (PRV) encodes a single immediate early gene called IE180. The IE180 protein is a potent transcriptional activator of viral genes involved in DNA replication and RNA transcription. A PRV mutant with both copies of IE180 deleted was constructed 20 years ago (S. Yamada and M. Shimizu, Virology 199:366–375, 1994, doi:10.1006/viro.1994.1134), but propagation of the mutant depended on complementing cell lines that expressed the toxic IE180 protein constitutively. Recently, Oyibo et al. constructed a novel set of PRV IE180 mutants and a stable cell line with inducible IE180 expression (H. Oyibo, P. Znamenskiy, H. V. Oviedo, L. W. Enquist, A. Zador, Front. Neuroanat. 8:86, 2014, doi:10.3389/fnana.2014.00086), which we characterized further here. These mutants failed to replicate new viral genomes, synthesize immediate early, early, or late viral proteins, and assemble infectious virions. The PRV IE180-null mutant did not form plaques in epithelial cell monolayers and could not spread from primary infected neurons to second-order neurons in culture. PRV IE180-null mutants lacked the property of superinfection exclusion. When PRV IE180-null mutants infected cells first, subsequent superinfecting viruses were not blocked in cell entry and formed replication compartments in epithelial cells, fibroblasts, and neurons. Cells infected with PRV IE180-null mutants survived as long as uninfected cells in culture while expressing a fluorescent reporter gene. Transcomplementation with IE180 in epithelial cells restored all mutant phenotypes to wild type. The conditional expression of PRV IE180 protein enables the propagation of replication-incompetent PRV IE180-null mutants and will facilitate construction of long-term single-cell-infecting PRV mutants for precise neural circuit tracing and high-capacity gene delivery vectors.

IMPORTANCE
Pseudorabies virus (PRV) is widely used for neural tracing in animal models. The virus replicates and spreads between synaptically connected neurons. Current tracing strains of PRV are cytotoxic and kill infected cells. Infected cells exclude superinfection with a second virus, limiting multiple virus infections in circuit tracing. By removing the only immediate early gene of PRV (called IE180), the mutant virus will not replicate or spread in epithelial cells, fibroblasts, or neurons. The wild-type phenotype can be restored by transcomplementation of infected cells with IE180. The PRV IE180-null mutant can express fluorescent reporters for weeks in cells with no toxicity; infected cells survive as long as uninfected cells. Infection with the mutant virus allows superinfection of the same cell with a second virus that can enter and replicate. The PRV IE180-null mutant will permit conditional long-term tracing in animals and is a high-capacity vector for gene delivery.
Superresolution Imaging Captures Carbohydrate Utilization Dynamics in Human Gut Symbionts
Authors: Karunatilaka Krishanthi S. , Cameron Elizabeth A. , Martens Eric C. , Koropatkin Nicole M. , Biteen Julie S. 
Published: November 2014

ABSTRACT

Gut microbes play a key role in human health and nutrition by catabolizing a wide variety of glycans via enzymatic activities that are not encoded in the human genome. The ability to recognize and process carbohydrates strongly influences the structure of the gut microbial community. While the effects of diet on the microbiota are well documented, little is known about the molecular processes driving metabolism. To provide mechanistic insight into carbohydrate catabolism in gut symbionts, we studied starch processing in real time in the model
Bacteroides thetaiotaomicron
starch utilization system (Sus) by single-molecule fluorescence. Although previous studies have explored Sus protein structure and function, the transient interactions, assembly, and collaboration of these outer membrane proteins have not yet been elucidated in live cells. Our live-cell superresolution imaging reveals that the polymeric starch substrate dynamically recruits Sus proteins, serving as an external scaffold for bacterial membrane assembly of the Sus complex, which may promote efficient capturing and degradation of starch. Furthermore, by simultaneously localizing multiple Sus outer membrane proteins on the
B. thetaiotaomicron
cell surface, we have characterized the dynamics and stoichiometry of starch-induced Sus complex assembly on the molecular scale. Finally, based on Sus protein knockout strains, we have discerned the mechanism of starch-induced Sus complex assembly in live anaerobic cells with nanometer-scale resolution. Our insights into the starch-induced outer membrane protein assembly central to this conserved nutrient uptake mechanism pave the way for the development of dietary or pharmaceutical therapies to control
Bacteroidetes
in the intestinal tract to enhance human health and treat disease.

IMPORTANCE
In this study, we used nanometer-scale superresolution imaging to reveal dynamic interactions between the proteins involved in starch processing by the prominent human gut symbiont
Bacteroides thetaiotaomicron
in real time in live cells. These results represent the first working model of starch utilization system (Sus) complex assembly and function during glycan catabolism and are likely to describe aspects of how other Sus-like systems function in human gut
Bacteroidetes
. Our results provide unique mechanistic insights into a glycan catabolism strategy that is prevalent within the human gut microbial community. Proper understanding of this conserved nutrient uptake mechanism is essential for the development of dietary or pharmaceutical therapies to control intestinal tract microbial populations, to enhance human health, and to treat disease.
Retrospective case review of missed opportunities for primary prevention of stroke and TIA in primary care: protocol paper
Authors: Moran Grace M , Calvert Melanie , Feltham Max G , Marshall Tom 
Published: November 2014

IntroductionStroke is a major health problem and transient ischaemic attack (TIA) is an important risk factor for stroke. Primary prevention of stroke and TIA will have the greatest impact on reducing the burden of these conditions. Evidence-based guidelines for stroke/TIA prevention identify individuals eligible for preventative interventions in primary care. This study will investigate: (1) the proportion of strokes/TIAs with prior missed opportunities for prevention in primary care; (2) the influence of patient characteristics on missed prevention opportunities and (3) how the proportion of missed prevention opportunities has changed over time.Methods and analysisA retrospective case review will identify first-ever stroke and patients with TIA between 2000 and 2013 using anonymised electronic medical records extracted from the health improvement network (THIN) database. Four categories of missed opportunities for stroke/TIA prevention will be sought: untreated high blood pressure in patients eligible for treatment (either blood pressure ≥160/100 or ≥140/90 mm Hg in patients at high cardiovascular disease (CVD) risk); patients with atrial fibrillation with high stroke risk and no anticoagulant therapy; no lipid modifying drug therapy prescribed in patients at high CVD risk or with familial hypercholesterolaemia. The proportion of patients with each missed opportunity and multiple missed opportunities will be calculated. Mixed effect logistic regression will model the relationship between demographic and patient characteristics and missed opportunities for care; practice will be included as a random effect.Ethics and disseminationTHIN data collection was approved by the NHS South East Multi-centre Research Ethics Committee (MREC) in 2003. This study was approved by the independent scientific review committee in May 2013. Dissemination of findings has the potential to change practice, improve the quality of care provided to patients and ultimately reduce the incidence of strokes and TIAs. Findings will be published in a peer-reviewed journal and disseminated at national and international conferences.
NOTCH Decoys That Selectively Block DLL/NOTCH or JAG/NOTCH Disrupt Angiogenesis by Unique Mechanisms to Inhibit Tumor Growth
Authors: Kangsamaksin Thaned , Murtomaki Aino , Kofler Natalie M. , Cuervo Henar , Chaudhri Reyhaan A. , Tattersall Ian W. , Rosenstiel Paul E. , Shawber Carrie J. , Kitajewski Jan 
Published: November 2014

Abstract
A proangiogenic role for Jagged (JAG)-dependent activation of NOTCH signaling in the endothelium has yet to be described. Using proteins that encoded different NOTCH1 EGF-like repeats, we identified unique regions of Delta-like ligand (DLL)–class and JAG-class ligand–receptor interactions, and developed NOTCH decoys that function as ligand-specific NOTCH inhibitors. N110–24 decoy blocked JAG1/JAG2–mediated NOTCH1 signaling, angiogenic sprouting in vitro, and retinal angiogenesis, demonstrating that JAG-dependent NOTCH signal activation promotes angiogenesis. In tumors, N110–24 decoy reduced angiogenic sprouting, vessel perfusion, pericyte coverage, and tumor growth. JAG–NOTCH signaling uniquely inhibited expression of antiangiogenic soluble (s) VEGFR1/sFLT1. N11–13 decoy interfered with DLL1–DLL4-mediated NOTCH1 signaling and caused endothelial hypersprouting in vitro, in retinal angiogenesis, and in tumors. Thus, blockade of JAG- or DLL-mediated NOTCH signaling inhibits angiogenesis by distinct mechanisms. JAG–NOTCH signaling positively regulates angiogenesis by suppressing sVEGFR1–sFLT1 and promoting mural–endothelial cell interactions. Blockade of JAG-class ligands represents a novel, viable therapeutic approach to block tumor angiogenesis and growth.
Significance: This is the first report identifying unique regions of the NOTCH1 extracellular domain that interact with JAG-class and DLL-class ligands. Using this knowledge, we developed therapeutic agents that block JAG-dependent NOTCH signaling and demonstrate for the first time that JAG blockade inhibits experimental tumor growth by targeting tumor angiogenesis. Cancer Discov; 5(2); 182–97. ©2014 AACR.
See related commentary by Briot and Iruela-Arispe, p. 112
This article is highlighted in the In This Issue feature, p. 97
Pericardial Endoscopy–Guided Left Atrial Appendage Ligation
Authors: Kimura Takehiro , Takatsuki Seiji , Miyoshi Shunichiro , Katsumata Yoshinori , Nishiyama Takahiko , Nishiyama Nobuhiro , Tanimoto Yoko , Aizawa Yoshiyasu , Jinzaki Masahiro , Fukuda Keiichi 
Published: November 2014

Background—
Approaches for closing the left atrial appendage (LAA) have been developed for stroke prevention. However, the prevailing maneuvers require an open-chest surgery, intravascular access, or transseptal puncture. We evaluated the feasibility and safety of pericardial endoscopy-guided LAA ligation in a canine model.

Methods and Results—
We used a total of 8 canines and computed tomography was performed before the procedures. After a double percutaneous pericardiocentesis, a transurethral rigid endoscope was inserted into the pericardial space. The ENDOLOOP ligature was advanced to the ostium of the LAA by counter pulling the tip of the LAA with forceps. After confirming the positioning guided by transesophageal echocardiography, the ligature was securely tightened. Acute success was evaluated by transesophageal echocardiography and chronic success was evaluated by blood testing, computed tomography, and transesophageal echocardiography. The LAA ligation was safely achieved in all canines without major complications. One month after the ligation, the ligated LAA was replaced by fibrotic tissue, and both the transesophageal echocardiography and computed tomographic images revealed no residual shunt. There was only a localized adhesion of the pericardium, where the original LAA was located, without the need for antibiotic or steroid administration. The postprocedural internal surface of the ligated LAA was smooth by virtue of intimal growth. Blood tests showed a slight elevation of the inflammatory markers, but this normalized spontaneously.

Conclusions—
Pericardial endoscopy-guided LAA ligation could provide an alternative, minimally invasive, and feasible solution for LAA closure that does not require vascular access or a transseptal puncture.
Long-Term Effects of 4 Popular Diets on Weight Loss and Cardiovascular Risk Factors
Authors: Atallah Renée , Filion Kristian B. , Wakil Susan M. , Genest Jacques , Joseph Lawrence , Poirier Paul , Rinfret Stéphane , Schiffrin Ernesto L. , Eisenberg Mark J. 
Published: November 2014

Background—
We conducted a systematic review to examine the efficacy of the Atkins, South Beach, Weight Watchers (WW), and Zone diets, with a particular focus on sustained weight loss at ≥12 months.

Methods and Results—

We systematically searched MEDLINE, EMBASE, and the Cochrane Library of Clinical Trials to identify randomized controlled trials (RCTs) published in English with follow-up ≥4 weeks that examined the effects of these 4 popular diets on weight loss and cardiovascular risk factors. We identified 12 RCTs (n=2559) with follow-up ≥12 months: 10 versus usual care (5 Atkins, 4 WW, and 1 South Beach) and 2 head-to-head (1 of Atkins, WW, and Zone, and 1 of Atkins, Zone, and control). At 12 months, the 10 RCTs comparing popular diets to usual care revealed that only WW was consistently more efficacious at reducing weight (range of mean changes: −3.5 to −6.0 kg versus −0.8 to −5.4 kg;
P
<0.05 for 3/4 RCTs). However, the 2 head-to-head RCTs suggest that Atkins (range: −2.1 to −4.7 kg), WW (−3.0 kg), Zone (−1.6 to −3.2 kg), and control (−2.2 kg) all achieved modest long-term weight loss. Twenty-four-month data suggest that weight lost with Atkins or WW is partially regained over time.

Conclusions—
Head-to-head RCTs, providing the most robust evidence available, demonstrated that Atkins, WW, and Zone achieved modest and similar long-term weight loss. Despite millions of dollars spent on popular commercial diets, data are conflicting and insufficient to identify one popular diet as being more beneficial than the others.
Functional Disability and Cognitive Impairment After Hospitalization for Myocardial Infarction and Stroke
Authors: Levine Deborah A. , Davydow Dimitry S. , Hough Catherine L. , Langa Kenneth M. , Rogers Mary A.M. , Iwashyna Theodore J. 
Published: November 2014

Background—
We assessed the acute and long-term effect of myocardial infarction (MI) and stroke on postevent functional disability and cognition while controlling for survivors’ changes in functioning over the years before the event.

Methods and Results—
Among participants in the nationally representative Health and Retirement Study with linked Medicare data (1998–2010), we determined within-person changes in functional limitations (basic and instrumental activities of daily living) and cognitive impairment after hospitalization for stroke (n=432) and MI (n=450), controlling for premorbid functioning using fixed-effects regression. In persons without baseline impairments, an acute MI yielded a mean acute increase of 0.41 functional limitations (95% confidence interval [CI], 0.18–0.63) with a linear increase of 0.14 limitations/year in the following decade. These increases were 0.65 limitations (95% CI, 0.07–1.23) and 0.27 limitations/year afterward for those with mild-to-moderate impairment at baseline. Stroke resulted in an acute increase of 2.07 (95% CI, 1.51–2.63) limitations because of the acute event and an increase of 0.15 limitations/year afterward for those unimpaired at baseline. There were 2.65 new limitations (95% CI, 1.86–3.44) and 0.19/year afterward for those with baseline mild-to-moderate impairment. Stroke hospitalization was associated with greater odds of moderate-to-severe cognitive impairment (odds ratio, 3.86; 95% CI, 2.10–7.11) at the time of the event, after adjustment for premorbid cognition but MI hospitalization was not.

Conclusions—
In this population-based cohort, most MI and stroke hospitalizations were associated with significant increases in functional disability at the time of the event and in the decade afterward. Survivors of MI and stroke warrant screening for functional disability over the long-term.
Reducing Door‐to‐Puncture Times for Intra‐Arterial Stroke Therapy: A Pilot Quality Improvement Project
Authors: Mehta Brijesh P. , Leslie‐Mazwi Thabele M. , Chandra Ronil V. , Bell Donnie L. , Sun Chung‐Huan J. , Hirsch Joshua A. , Rabinov James D. , Rost Natalia S. , Schwamm Lee H. , Goldstein Joshua N. , Levine Wilton C. , Gupta Rishi , Yoo Albert J. 
Published: November 2014

Background
Delays to intra‐arterial therapy (IAT) lead to worse outcomes in stroke patients with proximal occlusions. Little is known regarding the magnitude of, and reasons for, these delays. In a pilot quality improvement (QI) project, we sought to examine and improve our door‐puncture times.

Methods and Results

For anterior‐circulation stroke patients who underwent IAT, we retrospectively calculated in‐hospital time delays associated with various phases from patient arrival to groin puncture. We formulated and then implemented a process change targeted to the phase with the greatest delay. We examined the impact on time to treatment by comparing the pre‐ and post‐QI cohorts. One hundred forty‐six patients (93 pre‐ vs. 51 post‐QI) were analyzed. In the pre‐QI cohort (ie, sequential process), the greatest delay occurred from imaging to the neurointerventional (NI) suite (“picture‐suite”: median, 62 minutes; interquartile range [IQR], 40 to 82). A QI measure was instituted so that the NI team and anesthesiologist were assembled and the suite set up in parallel with completion of imaging and decision making. The post‐QI (ie, parallel process) median picture‐to‐suite time was 29 minutes (IQR, 21 to 41;
P
<0.0001). There was a 36‐minute reduction in median door‐to‐puncture time (143 vs. 107 minutes;
P
<0.0001). Parallel workflow and presentation during work hours were independent predictors of shorter door‐puncture times.

Conclusions
In‐hospital delays are a major obstacle to timely IAT. A simple approach for achieving substantial time savings is to mobilize the NI and anesthesia teams during patient evaluation and treatment decision making. This parallel workflow resulted in a >30‐minute (25%) reduction in median door‐to‐puncture times.
Synergistic Effect of Gestational Hypertension and Postpartum Incident Hypertension on Cardiovascular Health: A Nationwide Population Study
Authors: Yeh Jong Shiuan , Cheng Hao‐Min , Hsu Pai‐Feng , Sung Shih‐Hsien , Liu Wen‐Ling , Fang Hsin‐Ling , Chuang Shao‐Yuan 
Published: November 2014

Background

Gestational hypertension (
GH
) is a common complication of pregnancy and is associated with increased risk of incident hypertension in later life (
IH
) and cardiovascular events. However, the interactive effect of
GH
and
IH
on postpartum cardiovascular health remains unclear.

Methods and Results

A nationwide population‐based study was conducted using 1 million individuals from the Taiwan National Health Insurance database. Records from 1998 to 2009 were used to identify 1260 pregnant women with
GH
and without previous cardiovascular disease. The control group comprised 5040 pregnant women without
GH
, matched for age and date of delivery. During the follow‐up period (median duration, 5.8 years), 182 cardiovascular events developed. Women with
GH
had significantly higher risk of cardiovascular events (hazard ratio [95%
CI
], 2.44 [1.80 to 3.31]) and
IH
(8.29 [6.30 to 10.91]) than controls. Compared with women without
GH
and
IH
, there was a significantly higher risk of cardiovascular events for women without
GH
but with
IH
(relative risk [95%
CI
], 2.89 [1.27–6.58]), women with
GH
but without
IH
(1.66 [1.16–2.39]), and women with
GH
and
IH
(8.11 [5.36–12.30]). The synergy index was 2.91 (95%
CI
1.11 to 7.59), suggesting a positive interaction between
GH
and
IH
.

Conclusions

GH
increased the risk of subsequent
IH
. Women with both
GH
and
IH
were at a substantially higher cardiovascular risk than were women with either
GH
or
IH
. The synergistic adverse effect of
GH
and
IH
on postpartum cardiovascular health indicates that more attention should be paid to this special population.
Early Discharge After Primary Percutaneous Coronary Intervention: The Added Value of N‐Terminal Pro–Brain Natriuretic Peptide to the Zwolle Risk Score
Authors: Schellings Dirk A. A. M. , Adiyaman Ahmet , Giannitsis Evangelos , Hamm Christian , Suryapranata Harry , ten Berg Jurrien M. , Hoorntje Jan C. A. , van‘t Hof Arnoud W. J. 
Published: November 2014

Background

The Zwolle Risk Score (
ZRS
) identifies
ST
‐elevation myocardial infarction (
STEMI
) patients treated with primary percutaneous coronary intervention (
PPCI
) eligible for early discharge. We aimed to investigate whether baseline N‐terminal pro–brain natriuretic peptide (
NT
‐pro
BNP
) is also able to identify these patients and could improve future risk strategies.

Methods and Results

PPCI patients included in the Ongoing Tirofiban in Myocardial Infarction Evaluation (On‐TIME) II study were candidates (N=861). We analyzed whether ZRS and baseline NT‐proBNP predicted 30‐day mortality and assessed the occurrence of major adverse cardiac events (MACEs) and major bleeding. Receiver operating characteristic curve analysis was used to assess discriminative accuracy for ZRS, NT‐pro‐BNP, and their combination. After multiple imputation, 845 patients were included. Both ZRS >3 (hazard ratio [HR]=9.42;
P
<0.001) and log NT‐pro‐BNP (HR=2.61;
P
<0.001) values were associated with 30‐day mortality. On multivariate analysis, both the ZRS (HR=1.41; 95% confidence interval [CI]=1.27 to 1.56;
P
<0.001) and log NT‐proBNP (HR=2.09; 95% CI=1.59 to 2.74;
P
<0.001) independently predicted death at 30 days. The area under the curve for 30‐day mortality for combined ZRS/NT‐proBNP was 0.94 (95% CI=0.90 to 0.99), with optimal predictive values of a ZRS ≥2 and a NT‐proBNP value of ≥200 pg/
mL
. Using these cut‐off values, 64% of the study population could be identified as very low risk with zero mortality at 30 days follow‐up and low occurrence of MACEs and major bleeding between 48 hours and 10 days (1.3% and 0.6%, respectively).

Conclusion

Baseline
NT
‐pro
BNP
identifies a large group of low‐risk patients who may be eligible for early (48‐ to 72‐hour) discharge, whereas optimal predictive accuracy is reached by the combination of both baseline
NT
‐pro
BNP
and
ZRS
.