bluuebunny/crossref_metadata_2025_split huggingface dataset:
Flow Cytometry Analysis Using Sysmex UF-1000i Classifies Uropathogens Based on Bacterial, Leukocyte, and Erythrocyte Counts in Urine Specimens among Patients with Urinary Tract Infections
Authors: Monsen Tor , Rydén Patrik 
Published: December 2014

ABSTRACTUrinary tract infections (UTIs) are the second most common bacterial infection. Urine culture is the gold standard for diagnosis, but new techniques, such as flow cytometry analysis (FCA), have been introduced. The aim of the present study was to evaluate FCA characteristics regarding bacteriuria, leukocyturia, and erythrocyturia in relation to cultured uropathogens in specimens from patients with a suspected UTI. We also wanted to evaluate whether the FCA characteristics can identify uropathogens prior to culture. From a prospective study, 1,587 consecutive urine specimens underwent FCA prior to culture during January and February 2012. Outpatients and inpatients (79.6% and 19.4%, respectively) were included, of whom women represented 67.5%. In total, 620 specimens yielded growth, of whichEscherichia colirepresented 65%,Enterococcusspp. 8%,Klebsiellaspp. 7%, andStaphylococcusspp. 5%. For the uropathogens, the outcome of FCA was compared against the results for specimens withE. coliand those with a negative culture.E. colihad high bacterial (median, 17,914/μl), leukocyte (median, 348/μl), and erythrocyte (median, 23/μl) counts. With the exception ofKlebsiellaspp., the majority of the uropathogens had considerable or significantly lower bacterial counts than that ofE. coli. High leukocyte counts were found in specimens withStaphylococcus aureus,Proteus mirabilis,Pseudomonas aeruginosa, and group C streptococci. Elevated erythrocyte counts were found forP. vulgaris,P. aeruginosa, and group C streptococci, as well as forStaphylococcus saprophyticus. In essence, FCA adds new information about the bacterial, leukocyte, and erythrocyte counts in urine specimens for different uropathogens. Based on FCA characteristics, uropathogens can be classified and identified prior to culture.E. coliandKlebsiellaspp. have similar FCA characteristics.
Regulatory T Cells Resist Virus Infection-Induced Apoptosis
Authors: Che Jenny W. , Kraft Anke R. M. , Selin Liisa K. , Welsh Raymond M. 
Published: December 2014

ABSTRACTRegulatory T (Treg) cells are important in the maintenance of self-tolerance, and the depletion of Treg cells correlates with autoimmune development. It has been shown that type I interferon (IFN) responses induced early in the infection of mice can drive memory (CD44hi) CD8 and CD4 T cells into apoptosis, and we questioned here whether the apoptosis of CD44-expressing Treg cells might be involved in the infection-associated autoimmune development. Instead, we found that Treg cells were much more resistant to apoptosis than CD44hi CD8 and CD4 T cells at days 2 to 3 after lymphocytic choriomeningitis virus infection, when type I IFN levels are high. The infection caused a downregulation of the interleukin-7 (IL-7) receptor, needed for survival of conventional T cells, while increasing on Treg cells the expression of the high-affinity IL-2 receptor, needed for STAT5-dependent survival of Treg cells. The stably maintained Treg cells early during infection may explain the relatively low incidence of autoimmune manifestations among infected patients.IMPORTANCEAutoimmune diseases are controlled in part by regulatory T cells (Treg) and are thought to sometimes be initiated by viral infections. We tested the hypothesis that Treg may die off at early stages of infection, when virus-induced factors kill other lymphocyte types. Instead, we found that Treg resisted this cell death, perhaps reducing the tendency of viral infections to cause immune dysfunction and induce autoimmunity.
Live-Attenuated Measles Virus Vaccine Targets Dendritic Cells and Macrophages in Muscle of Nonhuman Primates
Authors: Rennick Linda J. , de Vries Rory D. , Carsillo Thomas J. , Lemon Ken , van Amerongen Geert , Ludlow Martin , Nguyen D. Tien , Yüksel Selma , Verburgh R. Joyce , Haddock Paula , McQuaid Stephen , Duprex W. Paul , de Swart Rik L. 
Published: December 2014

ABSTRACT

Although live-attenuated measles virus (MV) vaccines have been used successfully for over 50 years, the target cells that sustain virus replication
in vivo
are still unknown. We generated a reverse genetics system for the live-attenuated MV vaccine strain Edmonston-Zagreb (EZ), allowing recovery of recombinant (r)MV
EZ
. Three recombinant viruses were generated that contained the open reading frame encoding enhanced green fluorescent protein (EGFP) within an additional transcriptional unit (ATU) at various positions within the genome. rMV
EZ
EGFP(1), rMV
EZ
EGFP(3), and rMV
EZ
EGFP(6) contained the ATU upstream of the N gene, following the P gene, and following the H gene, respectively. The viruses were compared
in vitro
by growth curves, which indicated that rMV
EZ
EGFP(1) was overattenuated. Intratracheal infection of cynomolgus macaques with these recombinant viruses revealed differences in immunogenicity. rMV
EZ
EGFP(1) and rMV
EZ
EGFP(6) did not induce satisfactory serum antibody responses, whereas both
in vitro
and
in vivo
rMV
EZ
EGFP(3) was functionally equivalent to the commercial MV
EZ
-containing vaccine. Intramuscular vaccination of macaques with rMV
EZ
EGFP(3) resulted in the identification of EGFP
+
cells in the muscle at days 3, 5, and 7 postvaccination. Phenotypic characterization of these cells demonstrated that muscle cells were not infected and that dendritic cells and macrophages were the predominant target cells of live-attenuated MV.

IMPORTANCE
Even though MV strain Edmonston-Zagreb has long been used as a live-attenuated vaccine (LAV) to protect against measles, nothing is known about the primary cells in which the virus replicates
in vivo
. This is vital information given the push to move toward needle-free routes of vaccination, since vaccine virus replication is essential for vaccination efficacy. We have generated a number of recombinant MV strains expressing enhanced green fluorescent protein. The virus that best mimicked the nonrecombinant vaccine virus was formulated according to protocols for production of commercial vaccine virus batches, and was subsequently used to assess viral tropism in nonhuman primates. The virus primarily replicated in professional antigen-presenting cells, which may explain why this LAV is so immunogenic and efficacious.
TRIM5α Restriction Affects Clinical Outcome and Disease Progression in Simian Immunodeficiency Virus-Infected Rhesus Macaques
Authors: Wu Fan , Ourmanov Ilnour , Riddick Nadeene , Matsuda Kenta , Whitted Sonya , Plishka Ronald J. , Buckler-White Alicia , Starost Matthew F. , Hirsch Vanessa M. 
Published: December 2014

ABSTRACT
Tripartite motif-containing protein 5α (TRIM5α) is considered to be a potential target for cell-based gene modification therapy against human immunodeficiency virus type 1 (HIV-1) infection. In the present study, we used a relevant rhesus macaque model of infection with simian immunodeficiency virus from sooty mangabey (SIVsm) to evaluate the effect of TRIM5α restriction on clinical outcome. For macaques expressing a restrictive TRIM5 genotype, the disease outcomes of those infected with the wild-type TRIM-sensitive SIVsm strain and those infected with a virus with escape mutations in the capsid were compared. We found that TRIM5α restriction significantly delayed disease progression and improved the survival rate of SIV-infected macaques, supporting the feasibility of exploiting TRIM5α as a target for gene therapy against HIV-1. Furthermore, we also found that preservation of memory CD4 T cells was associated with protection by TRIM5α restriction, suggesting memory CD4 T cells or their progenitor cells as an ideal target for gene modification. Despite the significant effect of TRIM5α restriction on survival, SIV escape from TRIM5α restriction was also observed; therefore, this may not be an effective stand-alone strategy and may require combination with other targets.

IMPORTANCE
Recent studies suggest that it may be feasible not only to suppress viral replication with antiviral drugs but also potentially to eliminate or “cure” human immunodeficiency virus (HIV) infection. One approach being explored is the use of gene therapy to introduce genes that can restrict HIV replication, including a restrictive version of the host factor TRIM5α. TRIM5 was identified as a factor that restricts HIV replication in macaque cells. The rhesus gene is polymorphic, and some alleles are restrictive for primary SIVsm isolates, although escape mutations arise late in infection. Introduction of these escape mutations into the parental virus conferred resistance to TRIM5 on macaques. The present study evaluated these animals for long-term outcomes and found that TRIM5α restriction significantly delayed disease progression and improved the survival rate of SIV-infected macaques, suggesting that this could be a valid gene therapy approach that could be adapted for HIV.
Influenza Virus M2 Protein Ion Channel Activity Helps To Maintain Pandemic 2009 H1N1 Virus Hemagglutinin Fusion Competence during Transport to the Cell Surface
Authors: Alvarado-Facundo Esmeralda , Gao Yamei , Ribas-Aparicio Rosa María , Jiménez-Alberto Alicia , Weiss Carol D. , Wang Wei 
Published: December 2014

ABSTRACTThe influenza virus hemagglutinin (HA) envelope protein mediates virus entry by first binding to cell surface receptors and then fusing viral and endosomal membranes during endocytosis. Cleavage of the HA precursor (HA0) into a surface receptor-binding subunit (HA1) and a fusion-inducing transmembrane subunit (HA2) by host cell enzymes primes HA for fusion competence by repositioning the fusion peptide to the newly created N terminus of HA2. We previously reported that the influenza virus M2 protein enhances pandemic 2009 influenza A virus [(H1N1)pdm09] HA-pseudovirus infectivity, but the mechanism was unclear. In this study, using cell-cell fusion and HA-pseudovirus infectivity assays, we found that the ion channel function of M2 was required for enhancement of HA fusion and HA-pseudovirus infectivity. The M2 activity was needed only during HA biosynthesis, and proteolysis experiments indicated that M2 proton channel activity helped to protect (H1N1)pdm09 HA from premature conformational changes as it traversed low-pH compartments during transport to the cell surface. While M2 has previously been shown to protect avian influenza virus HA proteins of the H5 and H7 subtypes that have polybasic cleavage motifs, this study demonstrates that M2 can protect HA proteins from human H1N1 strains that lack a polybasic cleavage motif. This finding suggests that M2 proton channel activity may play a wider role in preserving HA fusion competence among a variety of HA subtypes, including HA proteins from emerging strains that may have reduced HA stability.IMPORTANCEInfluenza virus infects cells when the hemagglutinin (HA) surface protein undergoes irreversible pH-induced conformational changes after the virus is taken into the cell by endocytosis. HA fusion competence is primed when host cell enzymes cleave the HA precursor. The proton channel function of influenza virus M2 protein has previously been shown to protect avian influenza virus HA proteins that contain a polybasic cleavage site from pH-induced conformational changes during biosynthesis, but this effect is less well understood for human influenza virus HA proteins that lack polybasic cleavage sites. Using assays that focus on HA entry and fusion, we found that the M2 protein also protects (H1N1)pdm09 influenza A virus HA from premature conformational changes as it transits low-pH compartments during biosynthesis. This work suggests that M2 may play a wider role in preserving HA function in a variety of influenza virus subtypes that infect humans and may be especially important for HA proteins that are less stable.
The clinical characteristics of patients with glaucoma presenting to Botswana healthcare facilities: an observational study
Authors: Jackson Daniel J , Razai Mohammad S , Falama Rosemary , Mongwa Matlhogonolo , Mutapanduwa Mishell , Baemisi Chao , Josiah Engelinah , Nkomazana Oathokwa , Lehasa Alice , Brealey Evelyn , White Andrew J , Jankowski Deborah , Kerr-Muir Malcolm G , Martin Keith R , Ngondi Jeremiah M 
Published: December 2014

ObjectiveThis study aimed to establish the clinical characteristics of patients with glaucoma attending eye care facilities in Botswana, and management of glaucoma among patients who received care in these facilities. The study also aimed to calculate the number of new diagnoses of glaucoma within the glaucoma service.DesignA prospective, hospital-based, observational study.SettingA multicentre study was undertaken in government-run eye departments in Botswana from June to August 2012.ParticipantsAll patients with a diagnosis of glaucoma attending clinics at seven study sites were invited to participate.Outcome measuresExamination findings, diagnosis and management were extracted from individual patient-held medical charts. Sociodemographic characteristics, patient knowledge and understanding of glaucoma were assessed through face-to-face interviews. In addition, details of outpatient attendances for 2011 were collected from 21 government-run hospitals.ResultsThe majority of the 366 patients interviewed had a diagnosis of primary glaucoma (86.6%). The diagnoses were mainly made by ophthalmologists (48.6%) and ophthalmic nurses (44.0%). Many patients (38.5%) had been symptomatic for over 6 months before visiting an eye clinic. The mean presenting intraocular pressure was 28.2 mm Hg (SD 11.9 mm Hg). Most follow-up patients (79.2%) had not received surgery, however, many (89.5%) would accept surgery. Only 11.5% of participants had heard of glaucoma prior to diagnosis. Many participants (35.9%) did not understand glaucoma after being diagnosed. The majority (94.9%) of living first-degree relatives had never been examined. The number of newly diagnosed glaucoma cases for 2011 in the south of the country was 14.1/100 000; 95% CI (12.0 to 16.5), in the north it was 16.2/100 000; 95% CI (13.8 to 19.0).ConclusionsGlaucoma is a significant burden that presents challenges to ophthalmic services in Botswana. Many patients have limited understanding of the condition and poor access to services. There is a need to develop a treatment infrastructure to include safe surgery and a reliable supply of effective medication.
Construction and Analysis of the NCI-EDRN Breast Cancer Reference Set for Circulating Markers of Disease
Authors: Marks Jeffrey R. , Anderson Karen S. , Engstrom Paul , Godwin Andrew K. , Esserman Laura J. , Longton Gary , Iversen Edwin S. , Mathew Anu , Patriotis Christos , Pepe Margaret S. 
Published: December 2014

Abstract
Background: Many circulating biomarkers have been reported for the diagnosis of breast cancer, but few, if any, have undergone rigorous credentialing using prospective cohorts and blinded evaluation.
Methods: The NCI Early Detection Research Network (EDRN) has created a prospective, multicenter collection of plasma and serum samples from 832 subjects designed to evaluate circulating biomarkers for the detection and diagnosis of breast cancer. These samples are available to investigators who wish to evaluate their biomarkers using a set of blinded samples. The breast cancer reference set is composed of blood samples collected using a standard operating procedure at four U.S. medical centers from 2008 to 2010 from women undergoing either tissue diagnosis for breast cancer or routine screening mammography. The reference set contains samples from women with incident invasive cancer (n = 190), carcinoma in situ (n = 55), benign pathology with atypia (n = 63), benign disease with no atypia (n = 231), and women with no evidence of breast disease by screening mammography (BI-RADS 1 or 2, n = 276). Using a subset of plasma samples (n = 505) from the reference set, we analyzed 90 proteins by multiplexed immunoassays for their potential utility as diagnostic markers.
Results: We found that none of these markers is useful for distinguishing cancer from benign controls. However, elevated CA-125 does appear to be a candidate marker for estrogen receptor–negative cancers.
Conclusions: Markers that can distinguish benign breast conditions from invasive cancer have not yet been found.
Impact: Availability of prospectively collected samples should improve future validation efforts. Cancer Epidemiol Biomarkers Prev; 24(2); 435–41. ©2014 AACR.
Mitochondrial One-Carbon Metabolism Maintains Redox Balance during Hypoxia
Authors: Martínez-Reyes Inmaculada , Chandel Navdeep S. 
Published: December 2014

Abstract
Summary: Mitochondria generate high levels of reactive oxygen species (ROS) to activate protumorigenic signaling pathways. In parallel, the mitochondria must also increase their antioxidant capacity to lower ROS levels and prevent cancer cell death. In this issue of Cancer Discovery, Ye and colleagues demonstrate that serine catabolism through one-carbon metabolism within the mitochondrial matrix is necessary to maintain this redox balance. Cancer Discov; 4(12); 1371–3. ©2014 AACR.
See related article by Ye et al., p. 1406
Variable Clinical Features and Ablation of Manifest Nodofascicular/Ventricular Pathways
Authors: Hoffmayer Kurt S. , Lee Byron K. , Vedantham Vasanth , Bhimani Ashish A. , Cakulev Ivan T. , Mackall Judith A. , Sahadevan Jayakumar , Rho Robert W. , Scheinman Melvin M. 
Published: December 2014

Background—
Manifest nodofascicular/ventricular (NFV) pathways are rare.

Methods and Results—
From 2008 to 2013, 4 cases were identified with manifest NFV pathways from 3 centers. The clinical findings and ablation sites are reported. All 4 cases presented with a wide complex tachycardia but with different QRS morphologies. Case 1 showed a left bundle branch block/superior axis, case 2 showed a right bundle branch block/inferior axis, case 3 showed a left bundle branch block/inferior axis, and case 4 showed a narrow QRS tachycardia and a wide complex tachycardia with a left bundle branch block/inferior axis. Three of the 4 tachycardias had atrioventricular dissociation ruling out extranodal accessory pathways, including atriofascicular pathways. Programmed extrastimuli showed evidence of a decremental accessory pathway in 3 of the 4 cases. Coexisting tachycardia mechanisms were seen in 3 of the 4 cases (atrioventricular nodal reentry tachycardia [2] and atrioventricular reentrant tachycardia [1]). Ablation in the slow pathway region eliminated the NFV pathway in 3 (transient in 1) with the other responding to surgical closure of a large atrial septal defect. The NFV pathway was a critical part of the tachycardia circuit in 1 and proved to be a bystander in the other 3 cases.

Conclusions—
Manifest NFV pathways presented with variable QRS expression dependent on the ventricular insertion site and often coexisted with other tachycardia mechanisms (atrioventricular nodal reentry tachycardia and atrioventricular reentrant tachycardia). In most cases, the atrial insertion of the pathway was in or near the slow pathway region. The NFV pathways were either critical to the tachycardia circuit or served as bystanders.
Intravenous Home Inotropic Use Is Safe in Pediatric Patients Awaiting Transplantation
Authors: Birnbaum Brian F. , Simpson Kathleen E. , Boschert Traci A. , Zheng Jie , Wallendorf Michael J. , Schechtman Kenneth , Canter Charles E. 
Published: December 2014

Background—
Intravenous inotropic therapy can be used to support children awaiting heart transplantation. Although use of this therapy is discouraged in adults because of poor outcomes, its use in children, particularly outpatient, has had limited evaluation. We aimed to evaluate the safety and efficacy of this practice.

Methods and Results—
A retrospective analysis of an intent to treat protocol was completed on United Network for Organ Sharing status 1A patients discharged on inotropic therapy from 1999 until 2012. Intravenous inotropic therapy was initiated for cardiac symptoms not amenable to oral therapy. Patients who were not status 1A or required >1 inotrope were excluded. Efficacy was analyzed by time to first event: transplantation; readmission until transplantation; improvement leading to inotrope withdrawal; or death. Safety included analysis of infection rates, line malfunctions, temporary hospitalization, neurological events, and arrhythmias. One hundred six patients met inclusion criteria. The mean age was 10.1±6.4 years, 47% of patients had congenital heart disease, and 80% of these patients had single ventricle physiology. In patients without congenital heart disease, 53% had dilated cardiomyopathy, 91% of patients received milrinone, 85% of patients underwent transplantation, 8% of patients successfully weaned from support as outpatients, whereas 6% died. Fifty percent of patients were readmitted before transplantation or weaning from support, of which 64% required only 1 readmission. The majority of readmissions were for heart failure.

Conclusions—
Outpatient intravenous inotropic therapy can be safely used as a bridge to transplantation in pediatric patients. A minority of patients can discontinue inotropic therapy because of clinical improvement.
Drug-Coated Balloon Versus Standard Percutaneous Transluminal Angioplasty for the Treatment of Superficial Femoral and Popliteal Peripheral Artery Disease
Authors: Tepe Gunnar , Laird John , Schneider Peter , Brodmann Marianne , Krishnan Prakash , Micari Antonio , Metzger Christopher , Scheinert Dierk , Zeller Thomas , Cohen David J. , Snead David B. , Alexander Beaux , Landini Mario , Jaff Michael R. 
Published: December 2014

Background—
Drug-coated balloons (DCBs) have shown promise in improving the outcomes for patients with peripheral artery disease. We compared a paclitaxel-coated balloon with percutaneous transluminal angioplasty (PTA) for the treatment of symptomatic superficial femoral and popliteal artery disease.

Methods and Results—

The IN.PACT SFA Trial is a prospective, multicenter, single-blinded, randomized trial in which 331 patients with intermittent claudication or ischemic rest pain attributable to superficial femoral and popliteal peripheral artery disease were randomly assigned in a 2:1 ratio to treatment with DCB or PTA. The primary efficacy end point was primary patency, defined as freedom from restenosis or clinically driven target lesion revascularization at 12 months. Baseline characteristics were similar between the 2 groups. Mean lesion length and the percentage of total occlusions for the DCB and PTA arms were 8.94±4.89 and 8.81±5.12 cm (
P
=0.82) and 25.8% and 19.5% (
P
=0.22), respectively. DCB resulted in higher primary patency versus PTA (82.2% versus 52.4%;
P
<0.001). The rate of clinically driven target lesion revascularization was 2.4% in the DCB arm in comparison with 20.6% in the PTA arm (
P
<0.001). There was a low rate of vessel thrombosis in both arms (1.4% after DCB and 3.7% after PTA [
P
=0.10]). There were no device- or procedure-related deaths and no major amputations.

Conclusions—
In this prospective, multicenter, randomized trial, DCB was superior to PTA and had a favorable safety profile for the treatment of patients with symptomatic femoropopliteal peripheral artery disease.

Clinical Trial Registration—

URL:
http://www.clinicaltrials.gov
. Unique Identifiers: NCT01175850 and NCT01566461.