bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of Xpert MRSA/SA Nasal and MRSA/SA ELITe MGB Assays for Detection of the
mecA
Gene with Susceptibility Testing Methods for Determination of Methicillin Resistance in Staphylococcus aureus Isolates
Authors: Belmekki Mohamed , Mammeri Hedi , Hamdad Farida , Rousseau Florence , Canarelli Brigitte , Biendo Maurice
Published: July 2013

In a series of 82
Staphylococcus
strains isolated from culture, 100% were identified as
Staphylococcus aureus
by matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS); 99.9% (77/82) of them were resistant to benzylpenicillin, oxacillin, and cefoxitin, and 6.1% (5/82) were susceptible to methicillin. Xpert MRSA/SA assay results were concordant with the phenotypic results in 76.8% (63/82) of cases and discordant in 23.2% (19/82) of cases. The MRSA/SA ELITe MGB kit results were concordant with phenotypic results in 100% of the cases. When comparing the Xpert MRSA/SA assay results with the MRSA/SA ELITe MGB kit results, 78% (64/82) of the cases were concordant, while 22% (18/82) of the cases were discordant. No statistically significant differences were observed between the two techniques. The PCR protocol that was used to validate the results of these two methods gave the following results: 49 were conventional methicillin-resistant
S. aureus
(MRSA) isolates (
mecA
positive and

mecA
LGA251

negative), and 25 were phenotypic MRSA isolates (
mecA
negative and

mecA
LGA251

positive).
UV-Fixed-Thick-Blotch Preparation Improves Sensitivity of Auramine Staining
Authors: Cejudo-García M. A. , Munoz-Davila M. J. , Martinez-Lirola M. J.
Published: July 2013

We describe here a very simple modification of the auramine staining procedure based on preparation of a UV-fixed thick blotch which allowed us to reach an overall sensitivity of 0.82 (592 acid-fast bacillus [AFB]-positive specimens/722 initial respiratory specimens with positive mycobacterial culture) and sensitivities of 0.93 (526 AFB-positive specimens/564 culture-positive specimens) for
Mycobacterium tuberculosis
complex and 0.42 (66 AFB-positive specimens/158 culture-positive specimens) for nontuberculous mycobacteria.
High Coxiella burnetii DNA Load in Serum during Acute Q Fever Is Associated with Progression to a Serologic Profile Indicative of Chronic Q Fever
Authors: Wielders C. C. H. , Wijnbergen P. C. A. , Renders N. H. M. , Schellekens J. J. A. , Schneeberger P. M. , Wever P. C. , Hermans M. H. A.
Published: July 2013

PCR is very effective in diagnosing acute Q fever in the early stages of infection, when bacterial DNA is present in the bloodstream but antibodies have not yet developed. The objective of this study was to further analyze the diagnostic value of semiquantitative real-time PCR (qPCR) in diagnosing acute Q fever in an outbreak situation. At the Jeroen Bosch Hospital, in 2009, qPCR testing for
Coxiella burnetii
DNA was performed for 2,715 patients suspected of having acute Q fever (positive,
n
= 385; negative,
n
= 2,330). The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the qPCR assay were calculated for patients with negative qPCR results with a follow-up sample obtained within 14 days (
n
= 305) and qPCR-positive patients with at least one follow-up sample (
n
= 369). The correctness of the qPCR result was based on immunofluorescence assay results for samples submitted for qPCR and follow-up testing. The sensitivity of the Q fever qPCR assay was 92.2%, specificity 98.9%, PPV 99.2%, and NPV 89.8%. Patients who later developed serologic profiles indicative of chronic Q fever infection had significantly higher
C. burnetii
DNA loads during the acute phase than did patients who did not (
P
< 0.001). qPCR testing is a valuable tool for the diagnosis of acute Q fever and should be used in outbreak situations when the onset of symptoms is <15 days earlier. Special attention is needed in the follow-up monitoring of patients with high
C. burnetii
DNA loads during the acute phase, as this might be an indicator for the development of a serologic profile indicative of chronic infection.
Methylation of Human Papillomavirus Type 16 CpG Sites at E2-Binding Site 1 (E2BS1), E2BS2, and the Sp1-Binding Site in Cervical Cancer Samples as Determined by High-Resolution Melting Analysis–PCR
Authors: Jacquin Elise , Baraquin Alice , Ramanah Rajeev , Carcopino Xavier , Morel Adrien , Valmary-Degano Séverine , Bravo Ignacio G. , de Sanjosé Silvia , Riethmuller Didier , Mougin Christiane , Prétet Jean-Luc
Published: July 2013

High-risk (HR) human papillomavirus (HPV)-associated carcinogenesis is driven mainly by the overexpression of E7 and E6 oncoproteins following viral DNA integration and the concomitant loss of the E2 open reading frame (ORF). However, the integration of HR-HPV DNA is not systematically observed in cervical cancers. The E2 protein acts as a transcription factor that governs viral oncogene expression. The methylation of CpGs in the E2-binding sites (E2BSs) in the viral long control region abrogates E2 binding, thus impairing the E2-mediated regulation of E7/E6 transcription. Here, high-resolution melting (HRM)–PCR was developed to quantitatively analyze the methylation statuses of E2BS1, E2BS2, and the specificity protein 1 (Sp1)-binding site in 119 HPV16-positive cervical smears. This is a rapid assay that is suitable for the analysis of cervical samples. The proportion of cancer samples with methylated E2BS1, E2BS2, and Sp1-binding site CpGs was 47%, whereas the vast majority of samples diagnosed as being within normal limits, low-grade squamous intraepithelial lesions (LSIL), or high-grade squamous intraepithelial lesions (HSIL) harbored unmethylated CpGs. Methylation levels varied widely, since some cancer samples harbored up to 60% of methylated HPV16 genomes. A pyrosequencing approach was used as a confirmation test and highlighted that quantitative measurement of methylation can be achieved by HRM-PCR. Its prognostic value deserves to be investigated alone or in association with other biomarkers. The reliability of this single-tube assay offers great opportunities for the investigation of HPV16 methylation in other HPV-related cancers, such as head and neck cancers, which are a major public health burden.
Novel Method Based on “En Passant” Mutagenesis Coupled with a Gaussia Luciferase Reporter Assay for Studying the Combined Effects of Human Cytomegalovirus Mutations
Authors: Drouot Emilien , Piret Jocelyne , Boivin Guy
Published: July 2013

Human cytomegalovirus (HCMV) resistance to antivirals is a major problem in immunocompromised patients. Drug resistance is characterized by phenotypic testing or genotypic analysis of the phosphotransferase (UL97) and DNA polymerase (UL54) genes. However, genotypic assays require further characterization of unknown mutations in the drug resistance phenotype. Here, we describe a novel method for generating single or multiple mutations anywhere in the HCMV genome and for studying their effects on drug susceptibility. This method is based on cloning of the reference AD169 strain in a bacterial artificial chromosome and the use of “en passant” mutagenesis in bacteria to introduce mutations in recombinant HCMV without scar sequences. We also used this methodology to introduce the
Gaussia
luciferase reporter gene into the genome of the recombinant virus. To validate our system, the well-characterized single mutants with UL97 A594V and UL54 E756K mutations as well as the undescribed A594V/E756K double mutant were generated and their drug susceptibility profiles were determined by measuring the luciferase activity in cell culture supernatants. Drug susceptibility phenotypes for the A594V (8.2-fold increase in ganciclovir 50% effective concentration [EC
50
]) and E756K (1.9-, 3.9-, and 3.0-fold increases in ganciclovir, foscarnet, and cidofovir EC
50
s, respectively) mutants were similar to those previously reported, while the double mutant exhibited 10.8-, 4.1-, and 2.0-fold increases in ganciclovir, foscarnet, and cidofovir EC
50
s, respectively. The combination of the
Gaussia
luciferase reporter-based assay with the markerless “en passant” mutagenesis methodology constitutes an efficient system for studying phenotypes with single or multiple HCMV mutations.
Virion Factors That Target Daxx To Overcome Intrinsic Immunity
Authors: Schreiner Sabrina , Wodrich Harald
Published: July 2013

PML nuclear bodies and their associated functions are part of an intrinsic cellular mechanism aimed at maintaining transcriptional control over viral gene expression and preventing replication of invading viruses. To overcome these barriers, many viruses express early nonstructural, multifunctional proteins to support the viral replication cycle or modulate host immune responses. Virion proteins constituting the invading particle are traditionally investigated for their role in transport during entry or egress and in the assembly of new virions. The additional functions of virion proteins have largely been ignored, in contrast to those of their nonstructural counterparts. A number of recent reports suggest that several virion proteins may also play vital roles in gene activation processes, in particular by counteracting intrinsic immune mechanisms mediated by the PML nuclear body-associated cellular factors Daxx, ATRX, and Sp100. These virion proteins share several features with their more potent nonstructural counterparts, and they may serve to bridge the gap in the early phase of an infection until immediate early viral gene expression is established. In this review, we discuss how virion proteins are an integral part of gene regulation among several viral families and to what extent structural proteins of incoming virions may contribute to species barrier, latency, and oncogenesis.
Uncoupling of Influenza A Virus Transcription and Replication through Mutation of the Unpaired Adenosine in the Viral RNA Promoter
Authors: te Velthuis Aartjan J. W. , Turrell Lauren , Vreede Frank T. , Fodor Ervin
Published: July 2013

Transcription and replication of the influenza A virus RNA genome are mediated by the viral RNA polymerase from a promoter consisting of the partially base-paired 3′ and 5′ termini of viral genome segments. Here we show that transcription and replication can be uncoupled by mutation of an unpaired adenosine in the 5′ strand of the promoter. This residue is important for transcription but not replication by being essential for the cap-binding activity of the RNA polymerase.
The C-Terminal Domain of Chikungunya Virus nsP2 Independently Governs Viral RNA Replication, Cytopathicity, and Inhibition of Interferon Signaling
Authors: Fros Jelke J. , van der Maten Erika , Vlak Just M. , Pijlman Gorben P.
Published: July 2013

Alphavirus nonstructural protein 2 (nsP2) has pivotal roles in viral RNA replication, host cell shutoff, and inhibition of antiviral responses. Mutations that individually rendered other alphaviruses noncytopathic were introduced into chikungunya virus nsP2. Results show that (i) nsP2 mutation P718S only in combination with KR649AA or adaptive mutation D711G allowed noncytopathic replicon RNA replication, (ii) prohibiting nsP2 nuclear localization abrogates inhibition of antiviral interferon-induced JAK-STAT signaling, and (iii) nsP2 independently affects RNA replication, cytopathicity, and JAK-STAT signaling.
The Short Stalk Length of Highly Pathogenic Avian Influenza H5N1 Virus Neuraminidase Limits Transmission of Pandemic H1N1 Virus in Ferrets
Authors: Blumenkrantz Deena , Roberts Kim L. , Shelton Holly , Lycett Samantha , Barclay Wendy S.
Published: July 2013

H5N1 influenza viruses pose a pandemic threat but have not acquired the ability to support sustained transmission between mammals in nature. The restrictions to transmissibility of avian influenza viruses in mammals are multigenic, and overcoming them requires adaptations in hemagglutinin (HA) and PB2 genes. Here we propose that a further restriction to mammalian transmission of the majority of highly pathogenic avian influenza (HPAI) H5N1 viruses may be the short stalk length of the neuraminidase (NA) protein. This genetic feature is selected for when influenza viruses adapt to chickens. In our study, a recombinant virus with seven gene segments from a human isolate of the 2009 H1N1 pandemic combined with the NA gene from a typical chicken-adapted H5N1 virus with a short stalk did not support transmission by respiratory droplet between ferrets. This virus was also compromised in multicycle replication in cultures of human airway epithelial cells at 32°C. These defects correlated with a reduction in the ability of virus with a short-stalk NA to penetrate mucus and deaggregate virions. The deficiency in transmission and in cleavage of tethered substrates was overcome by increasing the stalk length of the NA protein. These observations suggest that H5N1 viruses that acquire a long-stalk NA through reassortment might be more likely to support transmission between humans. Phylogenetic analysis showed that reassortment with long-stalk NA occurred sporadically and as recently as 2011. However, all identified H5N1 viruses with a long-stalk NA lacked other mammalian adapting features and were thus several genetic steps away from becoming transmissible between humans.
The
HEX1
Gene of Fusarium graminearum Is Required for Fungal Asexual Reproduction and Pathogenesis and for Efficient Viral RNA Accumulation of Fusarium graminearum Virus 1
Authors: Son Moonil , Lee Kyung-Mi , Yu Jisuk , Kang Minji , Park Jin Man , Kwon Sun-Jung , Kim Kook-Hyung
Published: July 2013

The accumulation of viral RNA depends on many host cellular factors. The hexagonal peroxisome (Hex1) protein is a fungal protein that is highly expressed when the DK21 strain of
Fusarium graminearum
virus 1 (FgV1) infects its host, and Hex1 affects the accumulation of FgV1 RNA. The Hex1 protein is the major constituent of the Woronin body (WB), which is a peroxisome-derived electron-dense core organelle that seals the septal pore in response to hyphal wounding. To clarify the role of Hex1 and the WB in the relationship between FgV1 and
Fusarium graminearum
, we generated targeted gene deletion and overexpression mutants. Although neither
HEX1
gene deletion nor overexpression substantially affected vegetative growth, both changes reduced the production of asexual spores and reduced virulence on wheat spikelets in the absence of FgV1 infection. However, the vegetative growth of deletion and overexpression mutants was increased and decreased, respectively, upon FgV1 infection compared to that of an FgV1-infected wild-type isolate. Viral RNA accumulation was significantly decreased in deletion mutants but was significantly increased in overexpression mutants compared to the viral RNA accumulation in the virus-infected wild-type control. Overall, these data indicate that the
HEX1
gene plays a direct role in the asexual reproduction and virulence of
F. graminearum
and facilitates viral RNA accumulation in the FgV1-infected host fungus.
Exosomes Derived from Epstein-Barr Virus-Infected Cells Are Internalized via Caveola-Dependent Endocytosis and Promote Phenotypic Modulation in Target Cells
Authors: Nanbo Asuka , Kawanishi Eri , Yoshida Ryuji , Yoshiyama Hironori
Published: July 2013

Epstein-Barr virus (EBV), a human gammaherpesvirus, establishes a lifelong latent infection in B lymphocytes and epithelial cells following primary infection. Several lines of evidence suggest that exosomes derived from EBV-infected cells are internalized and transfer viral factors, including EBV-encoded latent membrane protein and microRNAs, to the recipient cells. However, the detailed mechanism by which exosomes are internalized and their physiological impact on the recipient cells are still poorly understood. In this study, we visualized the internalization of fluorescently labeled exosomes derived from EBV-uninfected and EBV-infected B cells of type I and type III latency into EBV-negative epithelial cells. In this way, we demonstrated that exosomes derived from all three cell types were internalized into the target cells in a similar fashion. Internalization of exosomes was significantly suppressed by treatment with an inhibitor of dynamin and also by the knockdown of caveolin-1. Labeled exosomes were colocalized with caveolae and subsequently trafficked through endocytic pathways. Moreover, we observed that exosomes derived from type III latency cells upregulated proliferation and expression of intercellular adhesion molecule 1 (ICAM-1) in the recipient cells more significantly than did those derived from EBV-negative and type I latency cells. We also identified the EBV latent membrane protein 1 (LMP1) gene as responsible for induction of ICAM-1 expression. Taken together, our data indicate that exosomes released from EBV-infected B cells are internalized via caveola-dependent endocytosis, which, in turn, contributes to phenotypic changes in the recipient cells through transferring one or more viral factors.
The Homologous Tripartite Viral RNA Polymerase of A/Swine/Korea/CT1204/2009(H1N2) Influenza Virus Synergistically Drives Efficient Replication and Promotes Respiratory Droplet Transmission in Ferrets
Authors: Pascua Philippe Noriel Q. , Song Min-Suk , Kwon Hyeok-Il , Lim Gyo-Jin , Kim Eun-Ha , Park Su-Jin , Lee Ok-Jun , Kim Chul-Joong , Webby Richard J. , Webster Robert G. , Choi Young-Ki
Published: July 2013

We previously reported that influenza A/swine/Korea/1204/2009(H1N2) virus was virulent and transmissible in ferrets in which the respiratory-droplet-transmissible virus (CT-Sw/1204) had acquired simultaneous hemagglutinin (HA
D225G
) and neuraminidase (NA
S315N
) mutations. Incorporating these mutations into the nonpathogenic A/swine/Korea/1130/2009(H1N2, Sw/1130) virus consequently altered pathogenicity and growth in animal models but could not establish efficient transmission or noticeable disease. We therefore exploited various reassortants of these two viruses to better understand and identify other viral factors responsible for pathogenicity, transmissibility, or both. We found that possession of the CT-Sw/1204 tripartite viral polymerase enhanced replicative ability and pathogenicity in mice more significantly than did expression of individual polymerase subunit proteins. In ferrets, homologous expression of viral RNA polymerase complex genes in the context of the mutant Sw/1130 carrying the HA
225G
and NA
315N
modifications induced optimal replication in the upper nasal and lower respiratory tracts and also promoted efficient aerosol transmission to respiratory droplet contact ferrets. These data show that the synergistic function of the tripartite polymerase gene complex of CT-Sw/1204 is critically important for virulence and transmission independent of the surface glycoproteins. Sequence comparison results reveal putative differences that are likely to be responsible for variation in disease. Our findings may help elucidate previously undefined viral factors that could expand the host range and disease severity induced by triple-reassortant swine viruses, including the A(H1N1)pdm09 virus, and therefore further justify the ongoing development of novel antiviral drugs targeting the viral polymerase complex subunits.
The Template Specificity of Bacteriophage Φ6 RNA Polymerase
Authors: Qiao Jian , Mindich Leonard
Published: July 2013

Bacteriophage Φ6 contains three double-stranded RNA (dsRNA) genomic segments, L, M, and S. The RNA is located inside a core particle composed of multiple copies of a major structural protein, an RNA-dependent RNA polymerase, a hexameric NTPase, and an auxiliary protein. The virion RNA polymerase in the core particle transcribes segments M and S
in vitro
. Segment L is transcribed poorly because its transcript starts with GU instead of GG found on segments S and M. Transcription
in vivo
is modified by the binding of host protein YajQ to the outside the core particle so that segment L is transcribed well. This mechanism is the determinant of the temporal control of gene expression in Φ6. Mutants of Φ6 have been isolated that are independent of YajQ for transcription of segment L. The mutations are found in the gene of the viral polymerase or the major capsid protein or both. These mutants are capable of transcribing segment L with the GU start or GA or GC. The same is found to be true when YajQ is added to wild-type particles. Minus-strand synthesis has restrictions that are different from that of plus-strand synthesis, and YajQ or mutations to independence do not modify minus-strand synthesis behavior. Purified polymerase P2 is able to transcribe dsRNA, but transcription behavior of segment L by both wild-type and mutant polymerases is different from that seen in capsid structures. Adding YajQ to purified polymerase does not change its transcription specificity.
Presentation Overrides Specificity: Probing the Plasticity of Alphaviral Proteolytic Activity through Mutational Analysis
Authors: Lulla Valeria , Karo-Astover Liis , Rausalu Kai , Merits Andres , Lulla Aleksei
Published: July 2013

Semliki Forest virus (genus
Alphavirus
) is an important model for studying regulated nonstructural (ns) polyprotein processing. In this study, we evaluated the strictness of the previously outlined cleavage rules, accounting for the timing and outcome of each of three cleavages within the ns polyprotein P1234, and assessed the significance of residues P6 to P4 within the cleavage sites using an alanine scanning approach. The processing of the 1/2 and 3/4 sites was most strongly affected following changes in residues P5 and P4, respectively. However, none of the mutations had a detectable effect on the processing of the 2/3 site. An analysis of recombinant viruses bearing combinations of mutations in cleavage sites revealed tolerance toward the cooccurrence of native and mutated cleavage sites within the same polyprotein, suggesting a remarkable plasticity of the protease recognition pocket. Even in a virus in which all of the cleavage sequences were replaced with alanines in the P6, P5, and P4 positions, the processing pattern was largely preserved, without leading to reversion of cleavage site mutations. Instead, the emergence of second-site mutations was identified, among which Q706R/L in nsP2 was confirmed to be associated with the recognition of the P4 position within the modified cleavage sites. Our results imply that the spatial arrangement of the viral replication complex inherently contributes to scissile-site presentation for the protease, alleviating stringent sequence recognition requirements yet ensuring the precision and the correct order of processing events. Obtaining a proper understanding of the consequences of cleavage site manipulations may provide new tools for taming alphaviruses.
Interaction of Herpes Simplex Virus ICP0 with ND10 Bodies: a Sequential Process of Adhesion, Fusion, and Retention
Authors: Gu Haidong , Zheng Yi , Roizman Bernard
Published: July 2013

On entry into the nucleus, herpes simplex virus 1 (HSV-1) DNA localizes to nuclear bodies known as ND10. Gene repression imposed by ND10 is released by a viral protein, ICP0, via degradation of the ND10 constituents promyelocytic leukemia protein (PML) and Sp100 and the subsequent dispersal of ND10 bodies. In order to understand the dynamic interaction between ICP0 and ND10, we carried out deletion mapping to identify the domains of ICP0 responsible for its association with ND10. Here, we report the following. (i) An ND10 entry signal (ND10-ES), located between residues 245 and 474, is required for ICP0 to penetrate and fuse with ND10. ICP0 lacking ND10-ES adheres to the surface of ND10 but fails to enter. (ii) In the absence of ND10-ES, the E3 ubiquitin ligase of ICP0 facilitates the transient adhesion of the truncated ICP0 to the ND10 surface, whereas the presence of ND10-ES in ICP0 renders ND10 fusion regardless of the E3 ligase activity. (iii) The C terminus of ICP0 is required for retention of ICP0 in ND10 but plays no role in the recruitment process. (iv) The adverse effects of an inactive RING domain on viral replication are partially reversed by deleting either ND10-ES or the C-terminal retention domain, suggesting that additional ICP0 functions require the release of ICP0 from ND10. Based on these results, we conclude that association of ICP0 and ND10 is a dynamic process, in which three sequential steps—adhesion, fusion, and retention—are adopted to stabilize the interaction. A faithful execution of these steps defines the ultimate productivity of the virus.
Association of the Vaccinia Virus A11 Protein with the Endoplasmic Reticulum and Crescent Precursors of Immature Virions
Authors: Maruri-Avidal Liliana , Weisberg Andrea S. , Moss Bernard
Published: July 2013

The apparent
de novo
formation of viral membranes within cytoplasmic factories is a mysterious, poorly understood first step in poxvirus morphogenesis. Genetic studies identified several viral proteins essential for membrane formation and the assembly of immature virus particles. Their repression results in abortive replication with the accumulation of dense masses of viroplasm. In the present study, we further characterized one of these proteins, A11, and investigated its association with cellular and viral membranes under normal and abortive replication conditions. We discovered that A11 colocalized in cytoplasmic factories with the endoplasmic reticulum (ER) and L2, another viral protein required for morphogenesis. Confocal microscopy and subcellular fractionation indicated that A11 was not membrane associated in uninfected cells, whereas L2 still colocalized with the ER. Cell-free transcription and translation experiments indicated that both A11 and L2 are tail-anchored proteins that associate posttranslationally with membranes and likely require specific cytoplasmic targeting chaperones. Transmission electron microscopy indicated that A11, like L2, associated with crescent membranes and immature virions during normal infection and with vesicles and tubules near masses of dense viroplasm during abortive infection in the absence of the A17 or A14 protein component of viral membranes. When the synthesis of A11 was repressed, “empty” immature-virion-like structures formed in addition to masses of viroplasm. The immature-virion-like structures were labeled with antibodies to A17 and to the D13 scaffold protein and were closely associated with calnexin-labeled ER. These studies revealed similarities and differences between A11 and L2, both of which may be involved in the recruitment of the ER for virus assembly.
Reversible retinopathy associated with oral deferasirox therapy
Authors: Walia Harpreet S , Yan Jiong
Published: July 2013

A 17-year-old girl with a history of sickle cell anaemia undergoing chronic blood transfusions and iron-chelation therapy presented for multiple ophthalmic examinations. During treatment with deferoxamine, her examination remained stable but 2 years after changing to deferasirox she presented with decreased visual acuity and only mild funduscopic changes. Marked electrophysiological abnormalities were also evident. After cessation of deferasirox, her visual acuity improved and electrophysiological responses improved. No prior reports of deferasirox-related retinopathy are available. We suggest that oral deferasirox caused a reversible retinopathy in our patient and clinicians be aware of this entity.
Actinomyces naeslundiiandEikenella corrodensas rare causes of liver abscesses
Authors: Jaqua Nathan Thomas , Smith Adam J , Shin Terry T , Jahanmir Jay
Published: July 2013

A 48-year-old man with an unremarkable medical history was admitted with vague conditions of fever, chills, myalgias and malaise. Physical examination was remarkable for only scleral icterus. Laboratory evaluation revealed elevated aminotransferases, alkaline phosphatase and bilirubin. Imaging demonstrated two masses in the right lobe of his liver, which were ultimately drained and cultures demonstratedActinomycesandEikenella. He continued to have fever on broad-spectrum antibiotics until catheter drainage of the abscesses was performed. He was eventually discharged in improved condition on amoxicillin-clavulanate. His aminotransferases, alkaline phosphatase and bilirubin continued to improve and he remained afebrile and asymptomatic. A repeat CT 2 months after discharge demonstrated resolution of the abscesses.ActinomycesandEikenellaare rare causes of liver abscesses and treatment requires drainage and an extended course of antibiotics. The polymicrobial character typical of liver abscesses makes antibiotic therapy challenging when cultures reveal rare organisms such asActinomycesandEikenella.
Brainstem tuberculosis
Authors: Demetriou George A
Published: July 2013

We present a case of a 38-year-old-man who presented with 1-week history of developing weakness of peripheral and cranial nerves. His MRI scan of the brain showed a large cavitating lesion at the brainstem and two further lesions of the right cerebral cortex and his CT chest showed features of old tuberculosis (TB). The identification of acid-fast bacilli was confirmed by analysis of bronchoalveolar lavage taken during bronchoscopy. He was started on anti-TB medications and repeat MRI 3 months later confirmed shrinkage of the cavitating lesion.
Epulis fissuratum: consequence of ill-fitting prosthesis
Authors: Mohan Ravi Prakash Sasankoti , Verma Sankalp , Singh Udita , Agarwal Neha
Published: July 2013

A poorly fitted prosthesis can give rise to a plethora of problems like pain, discomfort in mastication and speech and epulis fissuratum. Epulis fissuratum refers to reactive tissue response to excessive mechanical pressure imparted by the poor fit of prosthesis. In this article, we discuss a case of epulis fissuratum in a 69-year-old male patient.
Back pain in a previously healthy teenager
Authors: Fitzgerald Felicity , Howard Julia , Bailey Fiona , Soleimanian Sorrush
Published: July 2013

We present the case of a 14-year-old previously healthy boy who presented to his general practitioner with back pain and fever after rugby training. He was initially treated for suspected discitis but during the course of his admission he rapidly deteriorated and developed severe necrotising pneumonia. He was intubated, ventilated and transferred to a paediatric intensive care unit. Panton-Valentine leukocidinStaphylococcus aureuswas suspected and subsequently identified in blood cultures.
How do patients with end-stage ankle arthritis decide between two surgical treatments? A qualitative study
Authors: Zaidi Razi , Pfeil Michael , Macgregor Alexander J , Goldberg Andy
Published: July 2013

Objective

To examine how patients decide between ankle fusion and ankle replacement in end-stage ankle arthritis.

Design

Purposive patient selection, semistructured interviews, thematic analysis.

Setting

Royal National Orthopaedic Hospital, Stanmore, UK.

Participants

14 patients diagnosed with end-stage ankle osteoarthritis.

Results

We interviewed 6 men and 8 women with a mean age of 58 years (range 41–83). All had opted for surgery after failure of at least 6 months of conservative management, sequentially trading-off daily activities to limit the evolving pain. To decide between two offered treatments of ankle fusion and total ankle replacement (TAR), three major sources informed the patients’ decision-making process: their surgeon, peers and the internet. The treating surgeon was viewed as the most reliable and influential source of information. Information gleaned from other patients was also important, but with questionable reliability, as was information from the internet, both of which invariably required validation by the surgeon and in some cases the general practitioner.

Conclusions

Patients seek knowledge from a wealth of sources including the internet, web forums and other patients. While they leverage each of these sources to guide decision-making, the most important and influential factor in governing how patients decide on any particular surgical intervention is their surgeon. A high quality doctor–patient relationship, coupled with clear, balanced and complete information is essential to enable shared decision-making to become a standard model of care.
Prospective registration, bias risk and outcome-reporting bias in randomised clinical trials of traditional Chinese medicine: an empirical methodological study
Authors: Liu Jian-Ping , Han Mei , Li Xin-Xue , Mu Yu-Jie , Lewith George , Wang Yu-Yi , Witt Claudia M , Yang Guo-Yan , Manheimer Eric , Snellingen Torkel , Berman Brian , Gluud Christian
Published: July 2013

Background

Clinical trials on Traditional Chinese Medicine (TCM) should be registered in a publicly accessible international trial register and report on all outcomes. We systematically assessed and evaluated TCM trials in registries with their subsequent publications.

Objective

To describe the characteristics of TCM trials, estimate bias risk and outcome-reporting bias in clinical trials.

Data sources and study selection

Fifteen trial registries were searched from their inception to July 2012 to identify randomised trials on TCM including Chinese herbs, acupuncture and/or moxibustion, cupping, tuina, qigong, etc.

Data extraction

We extracted data including TCM specialty and treated disease/conditions from the registries and searched for subsequent publications in PubMed and Chinese databases. We compared information in the registries of completed trials with any publications focusing on study design, sample size, randomisation, bias risk including reporting bias from the register protocol.

Results

1096 registered randomised trials were identified evaluating TCM, of which 505 were completed studies (46.1%). The most frequent conditions were pain (13.3%), musculoskeletal (11.7%), nervous (8.7%), digestive (7.1%), circulatory (6.5%), respiratory (6.3%), mental and behavioural disorders (6.2%) and cancer (6.0%). The trial register data identified parallel, phase II/III randomised trials with sample size estimations and blinding, but limited information about randomisation (sequence generation and allocation concealment). Comparing trial registration data of 115 completed trials (22.8%) with their subsequent 136 publications, inconsistencies were identified in one or more of the following: sample size (11%), outcome assessor blinding (37.5%), primary outcomes (29%) and safety (28%) reporting.

Conclusions

Increasing numbers of clinical trials investigating a variety of TCM interventions have been registered in international trial registries. The study design of registered TCM trials has improved in estimating sample size, use of blinding and placebos. However, selective outcome reporting is widespread and similar to conventional medicine and therefore study conclusions should be interpreted with caution.
Effects of enzyme replacement therapy for cardiac-type Fabry patients with a Chinese hotspot late-onset Fabry mutation (IVS4+919G>A)
Authors: Lin Hsiang-Yu , Liu Hao-Chuan , Huang Yu-Hsiu , Liao Hsuan-Chieh , Hsu Ting-Rong , Shen Chia-I , Li Shao-Tzu , Li Cheng-Fang , Lee Li-Hong , Lee Pi-Chang , Huang Chun-Kai , Chiang Chuan-Chi , Lin Ching-Yuang , Lin Shuan-Pei , Niu Dau-Ming
Published: July 2013

Objective

Current studies of newborn screening for Fabry disease in Taiwan have revealed a remarkably high prevalence of cardiac-type Fabry disease with a Chinese hotspot late-onset Fabry mutation (IVS4+919G>A).

Design

Retrospective cohort study.

Setting

Tertiary medical centre.

Participants

21 patients with cardiac-type Fabry disease (15 men and 6 women) as well as 15 patients with classic Fabry disease (4 men and 11 women) treated with biweekly intravenous infusions of agalsidase β (1 mg/kg) or agalsidase α (0.2 mg/kg) for at least 6 months.

Outcome measures

These data were collected at the time before enzyme replacement therapy (ERT) began and followed up after ERT for at least 6 months, including patient demographics, medical history, parameter changes of cardiac status and renal functions, plasma globotriaosylsphingosine (lyso-Gb3) and Mainz Severity Score Index.

Results

After 6–39 months of ERT, plasma lyso-Gb3 was found to be reduced in 89% (17/19) and 93% (14/15) of patients with cardiac-type and classic Fabry disease, respectively, which indicated an improvement of disease severity. For patients with cardiac-type Fabry disease, echocardiography revealed the reduction or stabilisation of left ventricular mass index (LVMI), the thicknesses of intraventricular septum (IVS) and left posterior wall (LPW) in 83% (15/18), 83% (15/18) and 67% (12/18) of patients, respectively, as well as 77% (10/13), 73% (11/15) and 60% (9/15) for those with classic type. Most patients showed stable renal function after ERT. There were statistically significant improvements (p<0.05) between the data at baseline and those after ERT for values of plasma lyso-Gb3, LVMI, IVS, LPW and Mainz Severity Score Index. No severe clinical events were reported during the treatment.

Conclusions

ERT is beneficial and appears to be safe for Taiwanese patients with cardiac-type Fabry disease, as well as for those with the classic type.
A school-based education programme to reduce salt intake in children and their families (School-EduSalt): protocol of a cluster randomised controlled trial
Authors: He Feng J , Wu Yangfeng , Ma Jun , Feng Xiangxian , Wang Haijun , Zhang Jing , Lin Ching-Ping , Yuan Jianhui , Ma Yuan , Yang Yide , Yan Lijing L , Jan Stephen , Nowson Caryl , MacGregor Graham A
Published: July 2013

Introduction

The current salt intake is very high for children as well as adults in China. A reduction in salt intake is one of the most cost-effective measures to curb the rapidly growing disease burden attributed to blood pressure and cardiovascular disease in the Chinese population. A lower salt diet starting from childhood has the potential to prevent the development of such conditions. The School-EduSalt (School-basedEducation Programme to ReduceSalt) study aims to determine whether an education programme targeted at school children can lower salt intake in children and their families.

Methods and analysis

The study is designed as a cluster randomised controlled trial. The location is Changzhi, Shanxi province in northern China. The study population will consist of 28 primary schools with 280 children aged ≈11 years and 560 adult family members. Children in the intervention group will be educated on how to reduce salt intake. They will then be empowered to deliver the salt reduction message home to their families. In particular, children need to persuade the person who does the cooking to reduce the amount of salt used during food preparations. The duration of the intervention is one school term (≈4.5 months). The primary outcome is the difference between the intervention and the control group in the change in 24 h urinary sodium and the secondary outcome is the difference between the intervention and control group in the change of blood pressure. An economic evaluation will be undertaken to assess cost-effectiveness.

Ethics and dissemination

The study has been approved by The Queen Mary Research Ethics Committee (QMREC2012/81) and Peking University Health Science Centre IRB (IRB00001052-12072). Study findings will be disseminated widely through conference presentations and peer-reviewed publications.

Protocol Registration

Protocol Registered on ClinicalTrials.govNCT01821144.
How often do our liver core biopsies reach current definitions of adequacy?
Authors: Fryer Eve , Wang Lai Mun , Verrill Clare , Fleming Kenneth
Published: July 2013

Needle core biopsy is a key tool in diagnosis and assessment of many medical liver diseases, but there is evidence that the combination of small size of the specimen obtained and the patchy nature of many of these diseases can result in misdiagnosis or incorrect staging. The Royal College of Pathologists has therefore published guidelines for assessment of adequacy. To assess whether these guidelines were being observed, we reviewed cases reported in our department over a 15-year period. Results showed that only 19.8% of cores would be considered adequate, 56.4% were suboptimal and 23.8% were inadequate. We discuss the issues around recommendations on the minimum size of liver biopsies, potential factors limiting biopsy size and whether further refinement of the guidelines for adequacy is required.
Elevation of protein kinase Cα stimulates osteogenic differentiation of mesenchymal stem cells through the TAT-mediated protein transduction system
Authors: Hua Wei-Kai , Shiau Ya-Huei , Lee Oscar K. , Lin Wey-Jinq
Published: July 2013

Mesenchymal stem cells (MSCs) can differentiate toward various lineages, including the osteogenic lineage, and thus hold great potential for clinic purposes. By using pharmacological inhibitors, protein kinase C (PKC) signaling has been shown to either negatively or positively regulate differentiation of bone, however, due to the low transfection efficiency in MSCs, the role of individual PKC isoforms is still not fully understood. In this study, we established a TAT peptide-mediated transduction system that efficiently delivered PKCα proteins into MSCs in a non-invasive fashion. The increased PKCα protein levels significantly promoted osteogenic differentiation in the murine mesenchymal C3H10T1/2 cells and in primary MSCs from both human and mouse, as demonstrated by the enhanced activity of the osteoblast marker, alkaline phosphatase, and the enhanced expression of the key transcription factor runx2. Mineralization is an important functional indication for bone differentiation. Our results further showed that PKCα promoted expression of the important osteocalcin gene and the accumulation of calcium minerals. Taken together, this study provides direct evidence showing that PKCα positively regulates osteogenic differentiation and demonstrates that the TAT peptide-mediated method enables functional study of specific PKC isoforms in MSCs without using viral infection. This may promote the application of PKCs in therapeutic treatment.
Overexpression ofHelicobacter pyloriVacA N-terminal fragment induces proinflammatory cytokine expression and apoptosis in human monocytic cell line through activation of NF-κB
Authors: Luo Jing-Jing , Li Cun-Yan , Liu Sheng , Yu Wen , Tang Shuang-Yang , Cai Heng-Ling , Zhang Yan
Published: July 2013

Vacuolating cytotoxin (VacA) is an important virulence factor in the pathogenesis of Helicobacter pylori-related diseases. The aim of this study was to investigate the function of the amino-terminal 476 residue fragment (p52) of VacA and the possible molecular mechanisms responsible for its induction of proinflammatory cytokines secretion and apoptosis. Human acute monocytic leukemia cell line THP-1 was used as an in vitro model to study proinflammatory cytokines secretion and apoptosis induced by transfection of a recombinant plasmid encoding the amino-terminal 476 residue fragment (p52) of VacA. The results showed that VacA p52 overexpression induced the production of tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1β), nitric oxide, and reactive oxygen species in THP-1 cells in a time-dependent manner. VacA p52 overexpression also promoted THP-1 cells apoptosis. In addition, VacA p52 triggered the activation of nuclear factor kappa B (NF-κB), indicating a possible mechanism for its induction of proinflammatory cytokines secretion and cell apoptosis. Our study demonstrated that the induction of cytokines secretion and apoptosis by VacA p52 in THP-1 cells could be mediated through activation of nuclear factor kappa B.
Theoretical constraints on kinetic k-essence dark energy model in El-Nabulsi fractional action cosmology
Authors: Pasqua Antonio , Chattopadhyay Surajit
Published: July 2013

In this work, we considered an effective scalar field theory described by a Lagrangian with a noncanonical kinetic term, which leads to accelerated expansion in the present Universe and is known as k-essence in the framework of the fractional action cosmology recently introduced by El-Nabulsi. We have chosen a particular ansatz for the scale factor and the scalar field, in which both are described as a power-law of the time, t. We have studied the behavior of some cosmological quantities in other models to obtain some useful information about the model considered. We observed that the equation of state parameter, w, has decreasing behavior and it never crosses the phantom divide line (i.e., w = –1). Studying the statefinder pair {r, s} and {w, w′}, we observed that the model considered is able to obtain the ΛCDM phase of the Universe.
ELECTROPHYSIOLOGICAL RESPONSES FROM VAGUS NERVE STIMULATION IN RATS
Authors: MOLLET LIES , RAEDT ROBRECHT , DELBEKE JEAN , EL TAHRY RIËM , GRIMONPREZ ANNELIES , DAUWE INE , DE HERDT VEERLE , MEURS ALFRED , WADMAN WYTSE , BOON PAUL , VONCK KRISTL
Published: July 2013

The mechanism of action of vagus nerve stimulation (VNS) for pharmacoresistant epilepsy is unknown and the therapeutic outcome is highly variable. We investigated stimulation-induced vagus nerve electrophysiological responses in rats using various stimulation parameters. Conduction velocity, I 50, rheobase and chronaxie were calculated. We identified an early and late component corresponding to an afferent compound action potential (CAP) and a remote laryngeal motor-evoked potential (LMEP), respectively. The conduction velocity (CAP: 26.2 ± 1.4 m/s; LMEP: 32.4 ± 2.4 m/s) and I 50 (CAP: 2.4 ± 0.3 mA; LMEP: 1.8±0.2 mA) were significantly different for both components, the rheobase (CAP: 140±30 μA; LMEP: 110±26 μA) and chronaxie (CAP: 66±7 μs; LMEP: 73±9 μs) were not. Using a pulse of 10 μs, the CAP saturated between 4–5 mA. Our method can be used to record VNS-induced electrophysiological responses in rats and provides an objective biomarker for electrical stimulation with various parameters in an experimental set-up. Our findings are potentially useful for clinical purposes in the sense that combination of VNS and recording of vagal nerve CAPs may help clinicians to determine the individual optimal intensity required to fully activate fast-conducting afferent fibers.
A FUSION METHOD FOR PARTIAL FINGERPRINT RECOGNITION
Authors: CHEN FANGLIN , LI MING , ZHANG YI
Published: July 2013

Conventional algorithms for fingerprint recognition are mainly based on minutiae information. However, the small number of minutiae in partial fingerprints is still a challenge in fingerprint matching. In this paper, a novel algorithm is proposed to improve the performance of partial fingerprint matching. A simulation scheme was firstly proposed to construct a serial of partial fingerprints with different area. Then, the influence of the fingerprint area in partial fingerprint recognition is studied. By comparing the performance of partial fingerprint recognition with different fingerprint area, some useful conclusions can be drawn: (1) The decrease of the fingerprint area degrades the performance of partial fingerprint recognition; (2) When the fingerprint area decreases, the genuine matching scores will decrease, whereas the imposter matching scores will increase. Based on these observations, we proposed a fusion scheme based on modified support vector machine (SVM) to combine the area information for fingerprint recognition. Experimental result illustrates the effectiveness of the proposed method.
STRONG DECAYS OF FIRST RADIAL EXCITATIONS OF LIGHTEST VECTOR MESON STATES
Authors: MENGESHA ELIAS , BHATNAGAR SHASHANK
Published: July 2013

In this work, we have employed Bethe–Salpeter equation (BSE) under Covariant Instantaneous Ansatz (CIA) to study strong decays of first radial excitations, ω(1420) and ρ(1450) of lightest vector mesons, ω(782) and ρ(770) respectively, through the processes: ω(1420)→ρ+π and ρ(1450)→ω+π. The main motivation for this work was our intention to resolve the problems involved in calculations of triangle quark-loop diagrams as pointed out earlier in Ref. 1 which appear in processes such as h→h'+h" and h→h'+γ (attempted recently) in BSE under CIA, which gives rise to complexities in amplitudes making calculations difficult. We explicitly show the mathematical procedure for handling the process, h→h'+h" involving three hadron–quark vertex functions, which is more involved than that for h→h'+γ and leads to decay widths which are in rough agreement with data.