bluuebunny/crossref_metadata_2025_split huggingface dataset:
The use of intravesicular alteplase for thrombolysis in a dog with urinary bladder thrombi
Authors: Hooi Kimberly S. , Lemetayer Julie D. 
Published: July 2017

AbstractObjectiveTo describe the use of alteplase for intravesicular thrombolysis in a dog after development of urinary tract obstruction from a blood clot in the urinary bladder.Case SummaryA 5.8 kg, 6.5‐year‐old female neutered Bichon Frise was presented for signs of acute hematuria. A complete blood count (CBC) revealed marked thrombocytopenia and leukopenia, and nonregenerative anemia. Bone marrow aspirate cytology revealed mild hypercellularity, mild megakaryocytic hyperplasia, mildly left‐shifted erythroid maturation, and moderately left‐shifted myeloid maturation, suggesting ongoing recovery from an acute bone marrow insult. Thrombocytopenia and hematuria resolved concurrently; however, stranguria and oliguria developed acutely. Ultrasonography identified two large presumed thrombi within the urinary bladder. A urinary catheter was placed and 4 doses of 0.5 mg of alteplase diluted in 10 mL of 0.9% sodium chloride were instilled into the bladder with a 4‐hour dwell time at 12‐hour intervals. Prothombin and activated partial thromboplastin times were monitored during therapy and remained within normal limits. One thrombus was successfully dissolved after 48 hours of therapy and the remaining thrombus was reduced in size and was voided upon removal of the urinary catheter.New or Unique Information ProvidedThis report describes the use of alteplase in a dog for thrombolysis of intravesicular thrombi. In patients that develop intravesicular thrombi, intravesical instillation of alteplase can be considered as a method for dissolution of these thrombi.
A pilot study evaluating the prognostic utility of platelet indices in dogs with septic peritonitis
Authors: Llewellyn Efa A. , Todd Jeffrey M. , Sharkey Leslie C. , Rendahl Aaron 
Published: July 2017

AbstractObjectiveTo characterize platelet indices at time of diagnosis of septic peritonitis in dogs and to assess the relationship between platelet parameter data and survival to discharge in dogs treated surgically.DesignRetrospective, observational, descriptive pilot study from 2009 to 2014.SettingUniversity teaching hospital.AnimalsForty‐eight dogs diagnosed with septic peritonitis were included in this study. Thirty‐six dogs had surgical source control. Blood samples from 46 healthy control dogs were used for reference interval (RI) generation.InterventionsNone.Measurements and Main ResultsDogs with septic peritonitis had significantly increased mean values for mean platelet volume (MPV), plateletcrit (PCT), and platelet distribution width (PDW) with increased proportions of dogs having values above the RI compared to healthy dogs. A significantly increased proportion of dogs with septic peritonitis had platelet counts above (12.5%) and below (8.3%) the RI, with no significant difference in mean platelet count compared to healthy dogs. No significant differences in the mean platelet count, MPV, PCT, or PDW were found between survivors and nonsurvivors in dogs with surgical source control; however, dogs with MPV values above the RI had significantly increased mortality compared to dogs within the RI (P = 0.025). Values outside the RI for other platelet parameters were not associated with significant differences in mortality.ConclusionsDogs with septic peritonitis have increased frequency of thrombocytosis and thrombocytopenia with increased MPV, PCT, and PDW. An increased MPV may be a useful indicator of increased risk of mortality in dogs treated surgically.
N-Terminal Pfs230 Domain Produced in Baculovirus as a Biological Active Transmission-Blocking Vaccine Candidate
Authors: Lee Shwu-Maan , Wu Chia-Kuei , Plieskatt Jordan L. , Miura Kazutoyo , Hickey John M. , King C. Richter 
Published: July 2017

ABSTRACT

Transmission-blocking vaccines have the potential to accelerate malaria parasite elimination by inducing antibodies that block parasite transmission from humans to mosquitoes. Pfs230, a gametocyte surface protein involved in gamete function, has long been a promising candidate. Due to the large size (3,135 amino acids), complex domains, and repeating 6-cysteine (6-Cys) motifs with a multitude of disulfide bonds, the feasibility of expression of a full-length protein has been difficult. A priority focus, therefore, has been on the generation of single domains, including N-terminal fragments. Here we utilized a heterologous expression system, baculovirus, to produce an N-terminal domain of Pfs230 (Pfs230C1). Pfs230C1 (amino acids 443 to 731) with a polyhistidine affinity tag was expressed in Super Sf9 cells. Since the native host lacks glycosylation machinery, a single N585Q mutation was made to eliminate potential N-linked glycosylation. The expressed protein, purified by nickel affinity, ion exchange, and size exclusion chromatography to >90% purity, was present in monomeric form with an observed mass of 33,510 Da (matching oxidized form). Peptide mapping and disulfide analysis confirmed the proper formation of predicted disulfide bonds. Antibodies, generated against Pfs230C1 in mice, bound to the gametocyte in an immunofluorescence assay (IFA) and demonstrated functional activity in both the standard membrane feeding assay (SMFA) and the exflagellation assay (EXA). The biochemical, biophysical, and immunological results reported herein support the continued advancement of an N-terminal Pfs230 antigen (Pfs230C1) as a component of a transmission-blocking vaccine. Our results also support the continued use of the scalable baculovirus expression system for the generation of complex
Plasmodium
proteins.
Fusarium Keratitis in Germany
Authors: Walther Grit , Stasch Serena , Kaerger Kerstin , Hamprecht Axel , Roth Mathias , Cornely Oliver A. , Geerling Gerd , Mackenzie Colin R. , Kurzai Oliver , von Lilienfeld-Toal Marie 
Published: July 2017

ABSTRACT

Fusarium
keratitis is a destructive eye infection that is difficult to treat and results in poor outcome. In tropical and subtropical areas, the infection is relatively common and associated with trauma or chronic eye diseases. However, in recent years, an increased incidence has been reported in temperate climate regions. At the German National Reference Center, we have observed a steady increase in case numbers since 2014. Here, we present the first German case series of eye infections with
Fusarium
species. We identified
Fusarium
isolates from the eye or eye-related material from 22 patients in 2014 and 2015. Thirteen isolates belonged to the
Fusarium solani
species complex (FSSC), 6 isolates belonged to the
Fusarium oxysporum
species complex (FOSC), and three isolates belonged to the
Fusarium fujikuroi
species complex (FFSC). FSSC was isolated in 13 of 15 (85%) definite infections and FOSC in 3 of 4 (75%) definite contaminations. Furthermore, diagnosis from contact lens swabs or a culture of contact lens solution turned out to be highly unreliable. FSSC isolates differed from FOSC and FFSC by a distinctly higher MIC for terbinafine. Outcome was often adverse, with 10 patients requiring keratoplasty or enucleation. The use of natamycin as the most effective agent against keratitis caused by filamentous fungi was rare in Germany, possibly due to restricted availability. Keratitis caused by
Fusarium
spp. (usually FSSC) appears to be a relevant clinical problem in Germany, with the use of contact lenses as the predominant risk factor. Its outcome is often adverse.
Survival, Persistence, and Isolation of the Emerging Multidrug-Resistant Pathogenic Yeast Candida auris on a Plastic Health Care Surface
Authors: Welsh Rory M. , Bentz Meghan L. , Shams Alicia , Houston Hollis , Lyons Amanda , Rose Laura J. , Litvintseva Anastasia P. 
Published: July 2017

ABSTRACT

The emerging multidrug-resistant pathogenic yeast
Candida auris
represents a serious threat to global health. Unlike most other
Candida
species, this organism appears to be commonly transmitted within health care facilities and causes health care-associated outbreaks. To better understand the epidemiology of this emerging pathogen, we investigated the ability of
C. auris
to persist on plastic surfaces common in health care settings compared with that of
Candida parapsilosis
, a species known to colonize the skin and plastics. Specifically, we compiled comparative and quantitative data essential to understanding the vehicles of spread and the ability of both species to survive and persist on plastic surfaces under controlled conditions (25°C and 57% relative humidity), such as those found in health care settings. When a test suspension of 10
4
cells was applied and dried on plastic surfaces,
C. auris
remained viable for at least 14 days and
C. parapsilosis
for at least 28 days, as measured by CFU. However, survival measured by esterase activity was higher for
C. auris
than
C. parapsilosis
throughout the 28-day study. Given the notable length of time
Candida
species survive and persist outside their host, we developed methods to more effectively culture
C. auris
from patients and their environment. Using our enrichment protocol, public health laboratories and researchers can now readily isolate
C. auris
from complex microbial communities (such as patient skin, nasopharynx, and stool) as well as environmental biofilms, in order to better understand and prevent
C. auris
colonization and transmission.
Evaluation of the Rapid Polymyxin NP Test for Polymyxin B Resistance Detection Using Enterobacter cloacae and Enterobacter aerogenes Isolates
Authors: Simar Shelby , Sibley Diane , Ashcraft Deborah , Pankey George 
Published: July 2017

ABSTRACT

Polymyxin resistance is an increasing problem worldwide. Currently, determining susceptibility to polymyxins is problematic and lengthy. Polymyxins diffuse poorly into agar, potentially giving inaccurate disk diffusion and Etest results. A rapid screening test (2 h) for the detection of polymyxin resistance in
Enterobacteriaceae
, developed by P. Nordmann and L. Poirel (rapid polymyxin NP test) in 2016, detects glucose metabolization in the presence of polymyxin E (PE) and PB via pH-induced color change. The sensitivity and specificity were 99.3 and 95.4%, respectively, with results obtained in ≤2 h. Our goal was to evaluate this test using PB against larger numbers of
Enterobacter
. A total of 143 nonduplicate
Enterobacter
isolates (102
E. cloacae
complex, 41
E. aerogenes
) were tested, including 136 collected from Ochsner Health System patients from March to May 2016 and 7 previously determined PB-resistant
E. cloacae
isolates from JMI Laboratories. MICs were determined via broth microdilution. For the rapid polymyxin NP test, a color change from orange to yellow is positive; a weak/no color change is deemed negative after 4 h. Of 143
Enterobacter
isolates, 25 were determined to be PB resistant by broth microdilution (MIC > 2 μg/ml), including all 7 JMI isolates. Of these 25, 7 were positive by the rapid polymyxin NP test (included 3/7 JMI isolates). All 118 isolates determined to be PB susceptible by broth microdilution were NP test negative. The sensitivity and specificity for the rapid polymyxin NP test were 25 and 100%, respectively, compared to broth microdilution. Although the rapid polymyxin NP test is a much faster method (2 to 4 h) for polymyxin resistance determination compared to broth microdilution (16 to 20 h), our study indicates that it may be subject to limitations when testing
Enterobacter
.
Designing HIV Testing Algorithms Based on 2015 WHO Guidelines Using Data from Six Sites in Sub-Saharan Africa
Authors: Kosack Cara S. , Shanks Leslie , Beelaert Greet , Benson Tumwesigye , Savane Aboubacar , Ng'ang'a Anne , Bita André , Zahinda Jean-Paul B. N. , Fransen Katrien , Page Anne-Laure 
Published: July 2017

ABSTRACT
Our objective was to evaluate the performance of HIV testing algorithms based on WHO recommendations, using data from specimens collected at six HIV testing and counseling sites in sub-Saharan Africa (Conakry, Guinea; Kitgum and Arua, Uganda; Homa Bay, Kenya; Douala, Cameroon; Baraka, Democratic Republic of Congo). A total of 2,780 samples, including 1,306 HIV-positive samples, were included in the analysis. HIV testing algorithms were designed using Determine as a first test. Second and third rapid diagnostic tests (RDTs) were selected based on site-specific performance, adhering where possible to the WHO-recommended minimum requirements of ≥99% sensitivity and specificity. The threshold for specificity was reduced to 98% or 96% if necessary. We also simulated algorithms consisting of one RDT followed by a simple confirmatory assay. The positive predictive values (PPV) of the simulated algorithms ranged from 75.8% to 100% using strategies recommended for high-prevalence settings, 98.7% to 100% using strategies recommended for low-prevalence settings, and 98.1% to 100% using a rapid test followed by a simple confirmatory assay. Although we were able to design algorithms that met the recommended PPV of ≥99% in five of six sites using the applicable high-prevalence strategy, options were often very limited due to suboptimal performance of individual RDTs and to shared falsely reactive results. These results underscore the impact of the sequence of HIV tests and of shared false-reactivity data on algorithm performance. Where it is not possible to identify tests that meet WHO-recommended specifications, the low-prevalence strategy may be more suitable.
Role of Viral Hemorrhagic Septicemia Virus Matrix (M) Protein in Suppressing Host Transcription
Authors: Ke Qi , Weaver Wade , Pore Adam , Gorgoglione Bartolomeo , Wildschutte Julia Halo , Xiao Peng , Shepherd Brian S. , Spear Allyn , Malathi Krishnamurthy , Stepien Carol A. , Vakharia Vikram N. , Leaman Douglas W. 
Published: July 2017

ABSTRACT
Viral hemorrhagic septicemia virus (VHSV) is a pathogenic fish rhabdovirus found in discrete locales throughout the Northern Hemisphere. VHSV infection of fish cells leads to upregulation of the host's virus detection response, but the virus quickly suppresses interferon (IFN) production and antiviral gene expression. By systematically screening each of the six VHSV structural and nonstructural genes, we identified matrix protein (M) as the virus' most potent antihost protein. Only M of VHSV genotype IV sublineage b (VHSV-IVb) suppressed mitochondrial antiviral signaling protein (MAVS) and type I IFN-induced gene expression in a dose-dependent manner. M also suppressed the constitutively active simian virus 40 (SV40) promoter and globally decreased cellular RNA levels. Chromatin immunoprecipitation (ChIP) studies illustrated that M inhibited RNA polymerase II (RNAP II) recruitment to gene promoters and decreased RNAP II C-terminal domain (CTD) Ser2 phosphorylation during VHSV infection. However, transcription directed by RNAP I to III was suppressed by M. To identify regions of functional importance, M proteins from a variety of VHSV strains were tested in cell-based transcriptional inhibition assays. M of a particular VHSV-Ia strain, F1, was significantly less potent than IVb M at inhibiting SV40/luciferase (Luc) expression yet differed by just 4 amino acids. Mutation of D62 to alanine alone, or in combination with an E181-to-alanine mutation (D62A E181A), dramatically reduced the ability of IVb M to suppress host transcription. Introducing either M D62A or D62A E181A mutations into VHSV-IVb via reverse genetics resulted in viruses that replicated efficiently but exhibited less cytotoxicity and reduced antitranscriptional activities, implicating M as a primary regulator of cytopathicity and host transcriptional suppression.

IMPORTANCE
Viruses must suppress host antiviral responses to replicate and spread between hosts. In these studies, we identified the matrix protein of the deadly fish novirhabdovirus VHSV as a critical mediator of host suppression during infection. Our studies indicated that M alone could block cellular gene expression at very low expression levels. We identified several subtle mutations in M that were less potent at suppressing host transcription. When these mutations were engineered back into recombinant viruses, the resulting viruses replicated well but elicited less toxicity in infected cells and activated host innate immune responses more robustly. These data demonstrated that VHSV M plays an important role in mediating both virus-induced cell toxicity and viral replication. Our data suggest that its roles in these two processes can be separated to design effective attenuated viruses for vaccine candidates.
Effects of Inner Nuclear Membrane Proteins SUN1/UNC-84A and SUN2/UNC-84B on the Early Steps of HIV-1 Infection
Authors: Schaller Torsten , Bulli Lorenzo , Pollpeter Darja , Betancor Gilberto , Kutzner Juliane , Apolonia Luis , Herold Nikolas , Burk Robin , Malim Michael H. 
Published: July 2017

ABSTRACT

Human immunodeficiency virus type 1 (HIV-1) infection of dividing and nondividing cells involves regulatory interactions with the nuclear pore complex (NPC), followed by translocation to the nucleus and preferential integration into genomic areas in proximity to the inner nuclear membrane (INM). To identify host proteins that may contribute to these processes, we performed an overexpression screen of known membrane-associated NE proteins. We found that the integral transmembrane proteins SUN1/UNC84A and SUN2/UNC84B are potent or modest inhibitors of HIV-1 infection, respectively, and that suppression corresponds to defects in the accumulation of viral cDNA in the nucleus. While laboratory strains (HIV-1
NL4.3
and HIV-1
IIIB
) are sensitive to SUN1-mediated inhibition, the transmitted founder viruses RHPA and ZM247 are largely resistant. Using chimeric viruses, we identified the HIV-1 capsid (CA) protein as a major determinant of sensitivity to SUN1, and
in vitro
-assembled capsid-nucleocapsid (CANC) nanotubes captured SUN1 and SUN2 from cell lysates. Finally, we generated
SUN1
−/−
and
SUN2
−/−
cells by using CRISPR/Cas9 and found that the loss of SUN1 had no effect on HIV-1 infectivity, whereas the loss of SUN2 had a modest suppressive effect. Taken together, these observations suggest that SUN1 and SUN2 may function redundantly to modulate postentry, nuclear-associated steps of HIV-1 infection.

IMPORTANCE
HIV-1 causes more than 1 million deaths per year. The life cycle of HIV-1 has been studied extensively, yet important steps that occur between viral capsid release into the cytoplasm and the expression of viral genes remain elusive. We propose here that the INM components SUN1 and SUN2, two members of the linker of nucleoskeleton and cytoskeleton (LINC) complex, may interact with incoming HIV-1 replication complexes and affect key steps of infection. While overexpression of these proteins reduces HIV-1 infection, disruption of the individual
SUN2
and
SUN1
genes leads to a mild reduction or no effect on infectivity, respectively. We speculate that SUN1/SUN2 may function redundantly in early HIV-1 infection steps and therefore influence HIV-1 replication and pathogenesis.
Bam35 Tectivirus Intraviral Interaction Map Unveils New Function and Localization of Phage ORFan Proteins
Authors: Berjón-Otero Mónica , Lechuga Ana , Mehla Jitender , Uetz Peter , Salas Margarita , Redrejo-Rodríguez Modesto 
Published: July 2017

ABSTRACT

The family
Tectiviridae
comprises a group of tailless, icosahedral, membrane-containing bacteriophages that can be divided into two groups by their hosts, either Gram-negative or Gram-positive bacteria. While the first group is composed of PRD1 and nearly identical well-characterized lytic viruses, the second one includes more variable temperate phages, like GIL16 or Bam35, whose hosts are
Bacillus cereus
and related Gram-positive bacteria. In the genome of Bam35, nearly half of the 32 annotated open reading frames (ORFs) have no homologs in databases (ORFans), being putative proteins of unknown function, which hinders the understanding of their biology. With the aim of increasing knowledge about the viral proteome, we carried out a comprehensive yeast two-hybrid analysis of all the putative proteins encoded by the Bam35 genome. The resulting protein interactome comprised 76 unique interactions among 24 proteins, of which 12 have an unknown function. These results suggest that the P17 protein is the minor capsid protein of Bam35 and P24 is the penton protein, with the latter finding also being supported by iterative threading protein modeling. Moreover, the inner membrane transglycosylase protein P26 could have an additional structural role. We also detected interactions involving nonstructural proteins, such as the DNA-binding protein P1 and the genome terminal protein (P4), which was confirmed by coimmunoprecipitation of recombinant proteins. Altogether, our results provide a functional view of the Bam35 viral proteome, with a focus on the composition and organization of the viral particle.

IMPORTANCE
Tailless viruses of the family
Tectiviridae
can infect commensal and pathogenic Gram-positive and Gram-negative bacteria. Moreover, they have been proposed to be at the evolutionary origin of several groups of large eukaryotic DNA viruses and self-replicating plasmids. However, due to their ancient origin and complex diversity, many tectiviral proteins are ORFans of unknown function. Comprehensive protein-protein interaction (PPI) analysis of viral proteins can eventually disclose biological mechanisms and thus provide new insights into protein function unattainable by studying proteins one by one. Here we comprehensively describe intraviral PPIs among tectivirus Bam35 proteins determined using multivector yeast two-hybrid screening, and these PPIs were further supported by the results of coimmunoprecipitation assays and protein structural models. This approach allowed us to propose new functions for known proteins and hypothesize about the biological role of the localization of some viral ORFan proteins within the viral particle that will be helpful for understanding the biology of tectiviruses infecting Gram-positive bacteria.