bluuebunny/crossref_metadata_2025_split huggingface dataset:
Assessing transfusion competency in junior doctors: a retrospective cohort study
Authors: Graham Jane , Grant‐Casey John , Alston Robert , Baker Paul , Pendry Kate 
Published: June 2013

BackgroundThere is scope to further improve the safety of transfusion practice within the United Kingdom. This study aims to identify the current role of junior doctors in the transfusion process and to assess their competency to appropriately prescribe blood and blood products to patients.Study Design and MethodsTransfusion competency in junior doctors training in a single region was addressed through anonymized questionnaires assessing factual knowledge, personal reflection, and documented evidence of competency. Factual knowledge comprised 33 true‐false questions (competency score) covering indications for transfusion, special requirements, risks of transfusion, and guidelines for testing in transfusion. Background data on current practice and education in transfusion medicine were addressed using multiple‐choice and single‐response questions.ResultsA total of 787 newly qualified doctors, comprising 79% of first‐year (F1) and 62% of second‐year (F2) Foundation doctors, completed the assessment over a 3‐week period. There was no improvement in competency score between F1 and F2 doctors (p = 0.1). Competency scores correlated most strongly with undergraduate education in transfusion medicine and attendance at hospital induction (p < 0.01). Junior doctors had a high confidence level with regard to prescribing blood, although only 78% were aware they had been competency assessed against national standards.ConclusionJunior doctors are involved in sampling, prescribing, consenting, and documenting transfusion practice frequently enough to maintain competency. They are rarely involved in the collection, bedside checking, or administration of blood despite current curriculum requirements. There is scope to significantly improve both the training and the assessment of transfusion competency in doctors.
Alleles of the LAN blood group system: molecular and serologic investigations
Authors: Reid Marion E. , Hue‐Roye Kim , Huang Angela , Velliquette Randall W. , Tani Yoshihiko , Westhoff Connie M. , Lomas‐Francis Christine , Zelinski Teresa 
Published: June 2013

BackgroundAnti‐Lan has been implicated in hemolytic transfusion reactions and hemolytic disease of the fetus and newborn. The LAN blood group system is encoded by ABCB6, whose gene product, ABCB6, belongs to the ATP‐binding cassette (ABC) efflux transporter superfamily. The purpose of this study was to characterize additional alleles by analyzing DNA from 14 (13 unrelated) subjects whose red blood cells were serologically defined as Lan−, Lan+w/−, or Lan+w.Study Design and MethodsGenomic DNA was extracted from blood samples recovered from liquid nitrogen storage. Intronic primers flanking each of the ABCB6 coding exons were used for polymerase chain reaction amplification. Amplicons were sequenced and analyzed by standard methods.ResultsAmong the study subjects, we identified five alleles (one with a nonsense change, three with frameshifts, one with a missense change) that encode the Lan− phenotype and four alleles (with missense changes) encoding either Lan+w or Lan+w/− phenotypes.ConclusionsOf the nine alleles we identified, three were novel and six were previously documented in the dbSNP. Of these six, only one allele was previously associated with Lan negativity. To date, 19 ABCB6 alleles that encode Lan− or Lan+w/−, or Lan+w phenotypes have been described.
Intranasal Immunization of Lambs with Serine/Threonine Phosphatase 2A against Gastrointestinal Nematodes
Authors: Mohamed Fawzi Elshaima , Cruz Bustos Teresa , Gómez Samblas Mercedes , González-González Gloria , Solano Jenifer , González-Sánchez María Elena , De Pablos Luis Miguel , Corral-Caridad María Jesús , Cuquerella Montserrat , Osuna Antonio , Alunda José María 
Published: June 2013

ABSTRACTSeven 3-month-old, female, helminth-free lambs were immunized intranasally with three doses (1 mg total) of a recombinant part of the catalytic region of the serine/threonine phosphatase 2A (PP2Ar) (group 1 [G1]). In addition, four lambs were used as an adjuvant control group (G2), four as unimmunized, infected controls (G3), and four as unimmunized, uninfected controls (G4). Fifteen days after the last immunization, lambs from G1, G2, and G3 were challenged with 10,000 larval stage 3 (L3) organisms in a plurispecific nematode infection composed of ca. 40%Trichostrongylus colubriformis, 40%Haemonchus contortus, and 20%Teladorsagia circumcincta. All the lambs were clinically monitored throughout the experiment. Parasitological (fecal egg output and immunological response), biopathological (packed-cell volume and leukocyte and eosinophil counts), and zootechnical (live-weight gain) analyses were conducted. On day 105 of the experiment, all the animals were slaughtered and the adult worm population in their abomasa examined. Intranasal administration of PP2Ar with bacterial walls as an adjuvant elicited a strong immune response in the immunized lambs, as evidenced by their humoral immune response. Immunized animals and animals receiving the adjuvant shed significantly (P< 0.001) fewer numbers of parasites' eggs in their feces. The immunization significantly reduced the helminth burden in the abomasa by the end of the experiment (>68%), protection being provided against bothHaemonchusandTeladorsagia. Live-weight gain in the immunized lambs was similar to that in the uninfected controls versus the infected or adjuvanted animal groups. Our results suggest that heterologous immunization of ruminants by intranasal administration may be efficacious in the struggle to control gastrointestinal helminths in these livestock.
Rifampin Drug Resistance Tests for Tuberculosis: Challenging the Gold Standard
Authors: Van Deun Armand , Aung Kya J. M. , Bola Valentin , Lebeke Rossin , Hossain Mohamed Anwar , de Rijk Willem Bram , Rigouts Leen , Gumusboga Aysel , Torrea Gabriela , de Jong Bouke C. 
Published: June 2013

ABSTRACT

The rapid diagnosis of rifampin resistance is hampered by a reported insufficient specificity of molecular techniques for detection of
rpoB
mutations. Our objective for this study was to document the prevalence and prognostic value of
rpoB
mutations with unclear phenotypic resistance. The study design entailed sequencing directly from sputum of first failure or relapse patients without phenotypic selection and comparison of the standard retreatment regimen outcome, according to the mutation present. We found that among all
rpoB
mutations, the best-documented “disputed” rifampin resistance mutations (511Pro, 516Tyr, 526Asn, 526Leu, 533Pro, and 572Phe) made up 13.1% and 10.6% of all mutations in strains from Bangladesh and Kinshasa, respectively. Except for the 511Pro and 526Asn mutations, most of these strains with disputed mutations tested rifampin resistant in routine Löwenstein-Jensen medium proportion method drug susceptibility testing (DST; 78.7%), but significantly less than those with common, undisputed mutations (96.3%). With 63% of patients experiencing failure or relapse in both groups, there was no difference in outcome of first-line retreatment between patients carrying a strain with disputed versus common mutations. We conclude that rifampin resistance that is difficult to detect by the gold standard, phenotypic DST, is clinically and epidemiologically highly relevant. Sensitivity rather than specificity is imperfect with any rifampin DST method. Even at a low prevalence of rifampin resistance, a rifampin-resistant result issued by a competent laboratory may not warrant confirmation, although the absence of a necessity for confirmation needs to be confirmed for molecular results among new cases. However, a result of rifampin susceptibility should be questioned when suspicion is very high, and further DST using a different system (i.e., genotypic after phenotypic testing) would be fully justified.
Extra-Aortic Mycotic Aneurysm Due to Group A Streptococcus after Pharyngitis
Authors: Biswas Jason S. , Lyons Oliver T. A. , Bell Rachel E. , Price Nicholas 
Published: June 2013

ABSTRACT

Mycotic aneurysms, especially outside the aorta, are uncommon, with group A
Streptococcus
a particularly rare cause. We report a case of extra-aortic mycotic aneurysm following a sore throat without demonstrable bacteremia where identification of the pathological organism was made by molecular diagnostic techniques after a standard laboratory culture was negative.
Broad-Spectrum Biosensor Capable of Detecting and Identifying Diverse Bacterial and Candida Species in Blood
Authors: Metzgar David , Frinder Mark , Lovari Robert , Toleno Donna , Massire Christian , Blyn Lawrence B. , Ranken Raymond , Carolan Heather E. , Hall Thomas A. , Moore David , Hansen Christian J. , Sampath Rangarajan , Ecker David J. 
Published: June 2013

ABSTRACT

We describe an assay which uses broad-spectrum, conserved-site PCR paired with mass spectrometry analysis of amplicons (PCR/electrospray ionization-mass spectrometry [ESI-MS]) to detect and identify diverse bacterial and
Candida
species in uncultured specimens. The performance of the assay was characterized using whole-blood samples spiked with low titers of 64 bacterial species and 6
Candida
species representing the breadth of coverage of the assay. The assay had an average limit of detection of 100 CFU of bacteria or
Candida
per milliliter of blood, and all species tested yielded limits of detection between 20 and 500 CFU per milliliter. Over 99% of all detections yielded correct identifications, whether they were obtained at concentrations well above the limit of detection or at the lowest detectable concentrations. This study demonstrates the ability of broad-spectrum PCR/ESI-MS assays to detect and identify diverse organisms in complex natural matrices that contain high levels of background DNA.
Multicenter Clinical Evaluation of VRESelect Agar for Identification of Vancomycin-Resistant Enterococcus faecalis and Enterococcus faecium
Authors: Anderson Neil W. , Buchan Blake W. , Young Carol L. , Newton Duane W. , Brenke Connie , Lapsley Linda , Granato Paul A. , Ledeboer Nathan A. 
Published: June 2013

ABSTRACT

A chromogenic medium for identification of vancomycin-resistant
Enterococcus faecalis
and
Enterococcus faecium
, VRE
Select
, was compared to bile esculin azide agar with 6 μg/ml vancomycin (BEAV) for the isolation of vancomycin-resistant enterococci (VRE) from stool specimens. At 24 to 28 h, VRE
Select
demonstrated 98.7% (confidence interval [CI], 96.1 to 99.7%) sensitivity and 99.0% (CI, 98.0 to 99.6%) specificity versus 85.1% (CI, 79.8 to 89.5%) and 90.1% (CI, 79.8 to 89.5%) sensitivity and specificity, respectively, for BEAV.
Preliminary Laboratory Report of Fungal Infections Associated with Contaminated Methylprednisolone Injections
Authors: Lockhart Shawn R. , Pham Cau D. , Gade Lalitha , Iqbal Naureen , Scheel Christina M. , Cleveland Angela A. , Whitney Anne M. , Noble-Wang Judith , Chiller Tom M. , Park Benjamin J. , Litvintseva Anastasia P. , Brandt Mary E. 
Published: June 2013

ABSTRACT

In September 2012, the Centers for Disease Control and Prevention (CDC) initiated an outbreak investigation of fungal infections linked to injection of contaminated methylprednisolone acetate (MPA). Between 2 October 2012 and 14 February 2013, the CDC laboratory received 799 fungal isolates or human specimens, including cerebrospinal fluid (CSF), synovial fluid, and abscess tissue, from 469 case patients in 19 states. A novel broad-range PCR assay and DNA sequencing were used to evaluate these specimens. Although
Aspergillus fumigatus
was recovered from the index case,
Exserohilum rostratum
was the primary pathogen in this outbreak and was also confirmed from unopened MPA vials.
Exserohilum rostratum
was detected or confirmed in 191 specimens or isolates from 150 case patients, primarily from Michigan (
n
= 67 patients), Tennessee (
n
= 26), Virginia (
n
= 20), and Indiana (
n
= 16). Positive specimens from Michigan were primarily abscess tissues, while positive specimens from Tennessee, Virginia, and Indiana were primarily CSF.
E. rostratum
antifungal susceptibility MIC
50
and MIC
90
values were determined for voriconazole (1 and 2 μg/ml, respectively), itraconazole (0.5 and 1 μg/ml), posaconazole (0.5 and 1 μg/ml), isavuconazole (4 and 4 μg/ml), and amphotericin B (0.25 and 0.5 μg/ml). Thirteen other mold species were identified among case patients, and four other fungal genera were isolated from the implicated MPA vials. The clinical significance of these other fungal species remains under investigation. The laboratory response provided significant support to case confirmation, enabled linkage between clinical isolates and injected vials of MPA, and described significant features of the fungal agents involved in this large multistate outbreak.
Rifampin Resistance Missed in Automated Liquid Culture System for Mycobacterium tuberculosis Isolates with Specific
rpoB
Mutations
Authors: Rigouts Leen , Gumusboga Mourad , de Rijk Willem Bram , Nduwamahoro Elie , Uwizeye Cécile , de Jong Bouke , Van Deun Armand 
Published: June 2013

ABSTRACT

WHO-endorsed phenotypic drug susceptibility testing (DST) methods for
Mycobacterium tuberculosis
are assumed to be the gold standard for identifying rifampin (RMP) resistance. However, previous results indicated that low-level, yet probably clinically relevant, RMP resistance linked to specific
rpoB
mutations is easily missed by some growth-based methods. We aimed to compare the level of resistance detected on Löwenstein-Jensen (LJ) medium with resistance detected by the Bactec MGIT 960 automated DST (MGIT-DST) system for various
rpoB
mutants. Full agreement between LJ and MGIT-DST was observed for mutations located at codons 513 (Lys or Pro) and 531 (Leu, Trp), which were always resistant by both methods. For mutations 511Pro, 516Tyr, 533Pro, 572Phe, and several 526 mutations, LJ and MGIT results were highly discordant, with MGIT-DST failing to give a result or declaring the strains susceptible. Our data show that phenotypic RMP resistance testing of
M. tuberculosis
is not a binary phenomenon for some
rpoB
mutations and that the widely used automated MGIT 960 system is prone to miss some RMP resistance-conferring mutations, while careful DST on LJ missed hardly any. Given the association of these mutations with poor clinical outcome, our findings suggest that the gold standard for rifampin resistance should be reconsidered, in order to address the present confusion caused by discrepancies between phenotypic and genotypic results. The impacts of these mutations will depend on the frequency of their occurrence, which may vary from one setting to another.
Control the Host Cell Cycle: Viral Regulation of the Anaphase-Promoting Complex
Authors: Fehr Anthony R. , Yu Dong 
Published: June 2013

ABSTRACT
Viruses commonly manipulate cell cycle progression to create cellular conditions that are most beneficial to their replication. To accomplish this feat, viruses often target critical cell cycle regulators in order to have maximal effect with minimal input. One such master regulator is the large, multisubunit E3 ubiquitin ligase anaphase-promoting complex (APC) that targets effector proteins for ubiquitination and proteasome degradation. The APC is essential for cells to progress through anaphase, exit from mitosis, and prevent a premature entry into S phase. These far-reaching effects of the APC on the cell cycle are through its ability to target a number of substrates, including securin, cyclin A, cyclin B, thymidine kinase, geminin, and many others. Recent studies have identified several proteins from a number of viruses that can modulate APC activity by different mechanisms, highlighting the potential of the APC in driving viral replication or pathogenesis. Most notably, human cytomegalovirus (HCMV) protein pUL21a was recently identified to disable the APC via a novel mechanism by targeting APC subunits for degradation, both during virus infection and in isolation. Importantly, HCMV lacking both viral APC regulators is significantly attenuated, demonstrating the impact of the APC on a virus infection. Work in this field will likely lead to novel insights into viral replication and pathogenesis and APC function and identify novel antiviral and anticancer targets. Here we review viral mechanisms to regulate the APC, speculate on their roles during infection, and identify questions to be addressed in future studies.
Genetic Determinants of Reovirus Pathogenesis in a Murine Model of Respiratory Infection
Authors: Nygaard Rachel M. , Lahti Linse , Boehme Karl W. , Ikizler Mine , Doyle Joshua D. , Dermody Terence S. , Schiff Leslie A. 
Published: June 2013

ABSTRACT

Many viruses invade mucosal surfaces to establish infection in the host. Some viruses are restricted to mucosal surfaces, whereas others disseminate to sites of secondary replication. Studies of strain-specific differences in reovirus mucosal infection and systemic dissemination have enhanced an understanding of viral determinants and molecular mechanisms that regulate viral pathogenesis. After peroral inoculation, reovirus strain type 1 Lang replicates to high titers in the intestine and spreads systemically, whereas strain type 3 Dearing (T3D) does not. These differences segregate with the viral S1 gene segment, which encodes attachment protein σ1 and nonstructural protein σ1s. In this study, we define genetic determinants that regulate reovirus-induced pathology following intranasal inoculation and respiratory infection. We report that two laboratory isolates of T3D, T3D
C
and T3D
F
, differ in the capacity to replicate in the respiratory tract and spread systemically; the T3D
C
isolate replicates to higher titers in the lungs and disseminates, while T3D
F
does not. Two nucleotide polymorphisms in the S1 gene influence these differences, and both S1 gene products are involved. T3D
C
amino acid polymorphisms in the tail and head domains of σ1 protein influence the sensitivity of virions to protease-mediated loss of infectivity. The T3D
C
polymorphism at nucleotide 77, which leads to coding changes in both S1 gene products, promotes systemic dissemination from the respiratory tract. A σ1s-null virus produces lower titers in the lung after intranasal inoculation and disseminates less efficiently to sites of secondary replication. These findings provide new insights into mechanisms underlying reovirus replication in the respiratory tract and systemic spread from the lung.
Autocrine CCL3 and CCL4 Induced by the Oncoprotein LMP1 Promote Epstein-Barr Virus-Triggered B Cell Proliferation
Authors: Tsai Shu-Chun , Lin Sue-Jane , Lin Cheau-Jye , Chou Ya-Ching , Lin Jiun-Han , Yeh Te-Huei , Chen Mei-Ru , Huang Li-Min , Lu Meng-You , Huang Ya-Chi , Chen Huan-Yun , Tsai Ching-Hwa 
Published: June 2013

ABSTRACTEpstein-Barr virus (EBV) alters the regulation and expression of a variety of cytokines in its host cells to modulate host immune surveillance and facilitate viral persistence. Using cytokine antibody arrays, we found that, in addition to the cytokines reported previously, two chemotactic cytokines, CCL3 and CCL4, were induced in EBV-infected B cells and were expressed at high levels in all EBV-immortalized lymphoblastoid cell lines (LCLs). Furthermore, EBV latent membrane protein 1 (LMP1)-mediated Jun N-terminal protein kinase activation was responsible for upregulation of CCL3 and CCL4. Inhibition of CCL3 and CCL4 in LCLs using a short hairpin RNA approach or by neutralizing antibodies suppressed cell proliferation and caused apoptosis, indicating that autocrine CCL3 and CCL4 are required for LCL survival and growth. Importantly, significant amounts of CCL3 were detected in EBV-positive plasma from immunocompromised patients, suggesting that EBV modulates this chemokinein vivo. This study reveals the regulatory mechanism and a novel function of CCL3 and CCL4 in EBV-infected B cells. CCL3 might be useful as a therapeutic target in EBV-associated lymphoproliferative diseases and malignancies.
Caveolar Endocytosis Is Required for Human PSGL-1-Mediated Enterovirus 71 Infection
Authors: Lin Hsiang-Yin , Yang Ya-Ting , Yu Shu-Ling , Hsiao Kuang-Nan , Liu Chia-Chyi , Sia Charles , Chow Yen-Hung 
Published: June 2013

ABSTRACT
Enterovirus 71 (EV71) causes hand, foot, and mouth disease and severe neurological disorders in children. Human scavenger receptor class B member 2 (hSCARB2) and P-selectin glycoprotein ligand-1 (PSGL-1) are identified as receptors for EV71. The underling mechanism of PSGL-1-mediated EV71 entry remains unclear. The endocytosis required for EV71 entry were investigated in Jurkat T and mouse L929 cells constitutively expressing human PSGL-1 (PSGL-1-L929) or human rhabdomyosarcoma (RD) cells displaying high SCARB2 but no PSGL-1 by treatment of specific inhibitors or siRNA. We found that disruption of clathrin-dependent endocytosis prevented EV71 infection in RD cells, while there was no influence in Jurkat T and PSGL-1-L929 cells. Disturbing caveolar endocytosis by specific inhibitor or caveolin-1 siRNA in Jurkat T and PSGL-1-L929 cells significantly blocked EV71 infection, whereas it had no effect on EV71 infection in RD cells. Confocal immunofluorescence demonstrated caveola, and EV71 was directly colocalized. pH-dependent endosomal acidification and intact membrane cholesterol were important for EV71 infection, as judged by the pretreatment of inhibitors that abrogated the infection. A receptor-dominated endocytosis of EV71 infection was observed: PSGL-1 initiates caveola-dependent endocytosis and hSCARB2 activates clathrin-dependent endocytosis.
Capsid Antibodies to Different Adeno-Associated Virus Serotypes Bind Common Regions
Authors: Gurda Brittney L. , DiMattia Michael A. , Miller Edward B. , Bennett Antonette , McKenna Robert , Weichert Wendy S. , Nelson Christian D. , Chen Wei-jun , Muzyczka Nicholas , Olson Norman H. , Sinkovits Robert S. , Chiorini John A. , Zolotutkhin Sergei , Kozyreva Olga G. , Samulski R. Jude , Baker Timothy S. , Parrish Colin R. , Agbandje-McKenna Mavis 
Published: June 2013

ABSTRACT
Interactions between viruses and the host antibody immune response are critical in the development and control of disease, and antibodies are also known to interfere with the efficacy of viral vector-based gene delivery. The adeno-associated viruses (AAVs) being developed as vectors for corrective human gene delivery have shown promise in clinical trials, but preexisting antibodies are detrimental to successful outcomes. However, the antigenic epitopes on AAV capsids remain poorly characterized. Cryo-electron microscopy and three-dimensional image reconstruction were used to define the locations of epitopes to which monoclonal fragment antibodies (Fabs) against AAV1, AAV2, AAV5, and AAV6 bind. Pseudoatomic modeling showed that, in each serotype, Fabs bound to a limited number of sites near the protrusions surrounding the 3-fold axes of the T=1 icosahedral capsids. For the closely related AAV1 and AAV6, a common Fab exhibited substoichiometric binding, with one Fab bound, on average, between two of the three protrusions as a consequence of steric crowding. The other AAV Fabs saturated the capsid and bound to the walls of all 60 protrusions, with the footprint for the AAV5 antibody extending toward the 5-fold axis. The angle of incidence for each bound Fab on the AAVs varied and resulted in significant differences in how much of each viral capsid surface was occluded beyond the Fab footprints. The AAV-antibody interactions showed a common set of footprints that overlapped some known receptor-binding sites and transduction determinants, thus suggesting potential mechanisms for virus neutralization by the antibodies.