bluuebunny/crossref_metadata_2025 huggingface dataset:
Casein kinase 1‐epsilon deletion increases mu opioid receptor‐dependent behaviors and binge eating1
Authors: Goldberg L. R. , Kirkpatrick S. L. , Yazdani N. , Luttik K. P. , Lacki O. A. , Keith Babbs R. , Jenkins D. F. , Evan Johnson W. , Bryant C. D.
Published: June 2017

Genetic and pharmacological studies indicate that casein kinase 1 epsilon (Csnk1e) contributes to psychostimulant, opioid, and ethanol motivated behaviors. We previously used pharmacological inhibition to demonstrate that Csnk1e negatively regulates the locomotor stimulant properties of opioids and psychostimulants. Here, we tested the hypothesis that Csnk1e negatively regulates opioid and psychostimulant reward using genetic inhibition and the conditioned place preference assay in Csnk1e knockout mice. Similar to pharmacological inhibition, Csnk1e knockout mice showed enhanced opioid‐induced locomotor activity with the mu opioid receptor agonist fentanyl (0.2 mg/kg i.p.) as well as enhanced sensitivity to low‐dose fentanyl reward (0.05 mg/kg). Interestingly, female knockout mice also showed a markedly greater escalation in consumption of sweetened palatable food – a behavioral pattern consistent with binge eating that also depends on mu opioid receptor activation. No difference was observed in fentanyl analgesia in the 52.5°C hot plate assay (0–0.4 mg/kg), naloxone conditioned place aversion (4 mg/kg), or methamphetamine conditioned place preference (0–4 mg/kg). To identify molecular adaptations associated with increased drug and food behaviors in knockout mice, we completed transcriptome analysis via mRNA sequencing of the striatum. Enrichment analysis identified terms associated with myelination and axon guidance and pathway analysis identified a differentially expressed gene set predicted to be regulated by the Wnt signaling transcription factor, Tcf7l2. To summarize, Csnk1e deletion increased mu opioid receptor‐dependent behaviors, supporting previous studies indicating an endogenous negative regulatory role of Csnk1e in opioid behavior.
Biochemical characterization of LR769, a new recombinant factor VIIa bypassing agent produced in the milk of transgenic rabbits
Authors: Chevreux G. , Tilly N. , Leblanc Y. , Ramon C. , Faid V. , Martin M. , Dhainaut F. , Bihoreau N.
Published: June 2017

Background

The bypassing agent factor VII (FVIIa) is a first‐line therapy for the treatment of acute bleeding episodes in patients with haemophilia and high‐titre inhibitors. FVIIa is a highly post‐translationally modified protein that requires eukaryotic expression systems to produce a fully active molecule. A recombinant FVIIa was produced in the milk of transgenic rabbits to increase expression and provide an efficient, safe and affordable product after purification to homogeneity (LR769).

Aim

To present the biochemical and functional in vitro characteristics of LR769.

Results

Mass spectrometric analyses of the intact protein and of heavy and light chains revealed a fully activated, mature and properly post‐translationally modified protein notably regarding N/O‐glycosylations and γ‐carboxylation. Primary structure analysis, performed by peptide mapping, confirmed 100% of the sequence and the low level or absence of product‐derived impurities such as oxidized, deamidated and glycated forms. Low levels of aggregates and fragments were observed by different chromatographic methods. Higher order structure investigated by circular dichroism showed appropriate secondary/tertiary structures and conformational change in the presence of Ca2+ ions. Finally, activated partial thromboplastin time and thrombin generation assays showed the ability of LR769 to decrease coagulation time and to generate thrombin in haemophiliac‐A‐plasmas, even in the presence of inhibitors.

Conclusion

The innovative expression system used to produce LR769 yields a new safe and effective rhFVIIa for the treatment of haemophilia A or B patients with inhibitors.
Diagnostic value of STMN1, LMO2, HGAL, AID expression and 1p36 chromosomal abnormalities in primary cutaneous B cell lymphomas
Authors: Verdanet Evelyne , Dereure Olivier , René Céline , Tempier Ariane , Benammar‐Hafidi Assia , Gallo Mathieu , Frouin Eric , Durand Luc , Gazagne Isabelle , Costes‐Martineau Valérie , Cacheux Valère , Szablewski Vanessa
Published: June 2017

Aims

Distinction between primary cutaneous follicular lymphoma (PCFL) and primary cutaneous marginal zone lymphoma (PCMZL) is challenging, as clear‐cut immunophenotypical and cytogenetic criteria to segregate both entities are lacking.

Methods and results

To characterize PCFL and PCMZL more clearly and to define criteria helpful for the differential diagnosis, we compared expression of immunohistochemical markers [LIM‐only transcription factor 2 (LMO2), human germinal centre‐associated lymphoma (HGAL), stathmin 1 (STMN1), activation‐induced cytidine deaminase (AID), myeloid cell nuclear differentiation antigen (MNDA)] and the presence of cytogenetic abnormalities described previously in nodal follicular lymphoma [B cell lymphoma 2 (BCL2) and BCL6 breaks, 1p36 chromosomal region deletion (del 1p36)] in a series of 48 cutaneous follicular and marginal zone lymphomas [cutaneous follicular lymphoma (CFL) and cutaneous marginal zone lymphoma (CMZL)]. Immunostaining for STMN1, LMO2, HGAL and AID allowed the distinction between CFL and CMZL, and STMN1 was the most sensitive marker (100% CFL, 0% CMZL). LMO2, HGAL and AID were positive in 93.2%, 82.1% and 86.2% CFL (all CMZL‐negative). MNDA was expressed in both entities without significant difference (10.3% CFL, 30.8% CMZL, P = 0.18). BCL2, BCL6 breaks and the del 1p36 were present in 16.7%, 10.7% and 18.5% CFL and no CMZL. Finally, three and 29 CFL were reclassified as secondary cutaneous follicular lymphomas (SCFL) and PCFL without significant differences concerning phenotypical and cytogenetic features. BCL2, BCL6 breaks and the del 1p36 were present in 11.1%, 8% and 16.7% PCFL and did not impact the prognosis.

Conclusion

LMO2, HGAL, STMN1 and AID, but not MNDA, are discriminant for the recognition between CFL and CMZL. BCL2, BCL6 rearrangements and the del 1p36 have a role in the pathogenesis of PCFL, the latest being the most common alteration.
Investigation of adult stainless steel crown longevity as an interim restoration in pediatric patients
Authors: Discepolo Keri , Sultan Michael
Published: June 2017

Background

Preformed metal crowns, also known as stainless steel crowns (SSC), are utilized for providing full coverage to young permanent teeth and have been used extensively as restorations of primary teeth for approximately 65 years.

Aim

Evaluate permanent tooth SSC longevity as an interim restoration for teeth requiring full coverage restoration in pediatric patients.

Design

Retrospective chart review from 2006 through 2014 in a hospital‐based dental clinic for permanent tooth pre‐fabricated crown longevity. One hundred and fifty‐five SSCs were evaluated with regard to: age of patient at restoration placement, diagnoses, and duration of crown service. Duration service was further categorized as successful or failed.

Results

Of 155 SSCs, 137 were considered successfully functioning. Total failures were 18. The overall combined success rate for the study group was observed to be 88% with an average service period of 45.18 months. Significant success was noted in patients less than 9 years of age, and significant failure was observed in patients 12 years and older.

Conclusions

Stainless steel crowns are a viable option for interim restoration needs, with an 88% overall success rate during an average service period of 45.18 months.
Mission Congruence and Organization Design: An Empirical Analysis of Childcare Facilities
Authors: Barrenechea‐Mendez Marco A. , Ben‐Ner Avner
Published: June 2017

The paper investigates the relationship between mission congruence and the reliance on incentives and delegation of decision making, using a dataset on 206 childcare centers. We find that mission congruence between childcare teachers and employers is negatively related to the likelihood of reliance on financial incentives and positively related to teacher autonomy. These results support the idea that worker preferences play an important role in the design of organizations.
The impact of bank merger growth on CEO compensation
Authors: Chen Zhian , Hung Wing‐Yee , Li Donghui , Xing Lu
Published: June 2017

We examine the impact of bank mergers on chief executive officer (CEO) compensation during the period 1992–2014, a period characterised by significant banking consolidation. We show that CEO compensation is positively related to both merger growth and non‐merger internal growth, with the former relationship being higher in magnitude. While CEO pay–risk sensitivity is not significantly related to merger growth, CEO pay–performance sensitivity is negatively and significantly related to merger growth. Collectively, our results suggest that, through bank mergers, CEOs can earn higher compensation and decouple personal wealth from bank performance. Furthermore, we document a more severe agency problem in CEO compensation as a consequence of bank mergers relative to mergers in industrial firms. Finally, we find that the post‐financial crisis regulatory reform of executive compensation in banks has limited effectiveness in curbing the merger–pay links.
Impact of obesity on mortality in patients with diabetes: Meta‐analysis of 20 studies including 250,016 patients
Authors: Gao Fei , Wang Zhi Jian , Shen Hua , Yang Shi Wei , Nie Bin , Zhou Yu Jie
Published: June 2017

Aims/Introduction

The impact of body mass index on mortality among patients with diabetes remains controversial. Therefore, we carried out a meta‐analysis of pertinent studies.

Materials and Methods

We searched OVID/MEDLINE, EMBASE and Cochrane databases for all reported studies, which investigated the relationship between body mass index and mortality in patients with diabetes. Summary estimates of hazard ratios (HRs) were obtained with a random effects model. Univariate meta‐regressions were carried out.

Results

A total of 20 studies including 250,016 patients with diabetes were identified. The results of the present study showed a significantly reduced risk of all‐cause mortality in overweight patients (HR 0.82, 95% CI: 0.74–0.91, P < 0.0001, and I2 = 91.6%) as compared with normal weight patients. The survival benefits of obesity were only observed in the elderly patients (HR 0.69, 95% CI: 0.63–0.75, P < 0.0001, and I2 = 50.4%), but not in the younger patients (HR 1.01, 95% CI: 0.84–1.20, P = 0.96, I2 = 80.1%). Furthermore, the beneficial prognostic impacts on overweight (coefficient = 0.030, P = 0.041) and obesity (coefficient = 0.032, P = 0.010) were attenuated with clinical follow‐up duration.

Conclusions

The present meta‐analysis showed a significantly lower risk of all‐cause mortality in overweight patients with diabetes compared with normal weight patients. However, the survival benefits of obesity were only observed among the elderly patients.
Genetic architecture of acne vulgaris
Authors: Lichtenberger R. , Simpson M.A. , Smith C. , Barker J. , Navarini A.A.
Published: June 2017

Acne vulgaris is a ubiquitary skin disease characterized by chronic inflammation of the pilosebaceous unit resulting from bacterial colonization of hair follicles by Propionibacterium acnes, androgen‐induced increased sebum production, altered keratinization and inflammation. Here, we review our current understanding of the genetic architecture of this intriguing disease. We analysed genomewide association studies (GWAS) and candidate genes studies for acne vulgaris. Moreover, we included GWAS studies for the associated disease polycystic ovary syndrome (PCOS). Overall, the available data revealed sixteen genetic loci flagged by single nucleotide polymorphisms (SNPs), none of which has been confirmed yet by independent studies. Moreover, a GWAS for PCOS identified 21 susceptible loci. The genetic architecture is complex which has been revealed by GWAS. Further and larger studies in different populations are required to confirm or disprove results from candidate gene studies as well to identify signals that may overlap between different populations. Finally, studies on rare genetic variants in acne and associated diseases like PCOS may deepen our understanding of its pathogenesis.
Application of an advanced maximum likelihood estimation restoration method for enhanced‐resolution and contrast in second‐harmonic generation microscopy
Authors: SIVAGURU MAYANDI , KABIR MOHAMMAD M. , GARTIA MANAS RANJAN , BIGGS DAVID S.C. , SIVAGURU BARGHAV S. , SIVAGURU VIGNESH A. , FRIED GLENN A. , LIU GANG LOGAN , SADAYAPPAN SAKTHIVEL , TOUSSAINT KIMANI C.
Published: June 2017

Second‐harmonic generation (SHG) microscopy has gained popularity because of its ability to perform submicron, label‐free imaging of noncentrosymmetric biological structures, such as fibrillar collagen in the extracellular matrix environment of various organs with high contrast and specificity. Because SHG is a two‐photon coherent scattering process, it is difficult to define a point spread function (PSF) for this modality. Hence, compared to incoherent two‐photon processes like two‐photon fluorescence, it is challenging to apply the various PSF‐engineering methods to improve the spatial resolution to be close to the diffraction limit. Using a synthetic PSF and application of an advanced maximum likelihood estimation (AdvMLE) deconvolution algorithm, we demonstrate restoration of the spatial resolution in SHG images to that closer to the theoretical diffraction limit. The AdvMLE algorithm adaptively and iteratively develops a PSF for the supplied image and succeeds in improving the signal to noise ratio (SNR) for images where the SHG signals are derived from various sources such as collagen in tendon and myosin in heart sarcomere. Approximately 3.5 times improvement in SNR is observed for tissue images at depths of up to ∼480 nm, which helps in revealing the underlying helical structures in collagen fibres with an ∼26% improvement in the amplitude contrast in a fibre pitch. Our approach could be adapted to noisy and low resolution modalities such as micro‐nano CT and MRI, impacting precision of diagnosis and treatment of human diseases.
Ultraviolet A photosensitivity profile of dexchlorpheniramine maleate and promethazine‐based creams: Anti‐inflammatory, antihistaminic, and skin barrier protection properties
Authors: Facchini Gustavo , Eberlin Samara , Clerici Stefano Piatto , Alves Pinheiro Ana Lucia Tabarini , Costa Adilson
Published: June 2017

Background

Unwanted side effects such as dryness, hypersensitivity, and cutaneous photosensitivity are challenge for adherence and therapeutical success for patients using treatments for inflammatory and allergic skin response.

Aims

In this study, we compared the effects of two dermatological formulations, which are used in inflammatory and/or allergic skin conditions: dexchlorpheniramine maleate (DCP; 10 mg/g) and promethazine (PTZ; 20 mg/g).

Methods

We evaluated both formulations for phototoxicity potential, skin irritation, anti‐inflammatory and antihistaminic abilities, and skin barrier repair in vitro and ex vivo using the standard OECD test guideline n° 432, the ECVAM protocol n° 78, and cultured skin explants from a healthy patient. Ultraviolet A was chosen as exogenous agent to induce allergic and inflammatory response.

Results

Both PTZ and DCP promoted increases in interleukin‐1 (IL‐1) synthesis in response to ultraviolet A (UVA) radiation compared to control. However, the increase observed with PTZ was significantly greater than the DCP, indicating that the latter has a lower irritant potential. DCP also demonstrated a protective effect on UVA‐induced leukotriene B4 and nuclear factor kappa B (NF‐κB) synthesis. Conversely, PTZ demonstrates more robust UVA antihistaminic activity. Likewise, PTZ promoted a significantly greater increase in the production of involucrin and keratin 14, both associated with protective skin barrier property.

Conclusion

In conclusion, these data suggest possible diverging UVA response mechanisms of DCP and PTZ, which gives greater insight into the contrasting photosensitizing potential between DCP and PTZ observed in the patients.
Sleep hygiene behaviours in Iranian adolescents: an application of the Theory of Planned Behavior
Authors: Strong Carol , Lin Chung‐Ying , Jalilolghadr Shabnam , Updegraff John A. , Broström Anders , Pakpour Amir H.
Published: June 2017

Poor sleep quality and inadequate sleep in adolescents are a rising trend globally. The Theory of Planned Behaviour (TPB)—which centres on an individual's attitude toward performing the behaviour, subjective norms and perceived behavioural control—has been applied to examine sleep hygiene behaviours in young adults. We expanded on prior works by using a longitudinal design to examine the effects of TPB factors, together with sleep hygiene knowledge and planning constructs, on sleep hygiene behaviours and on sleep quality and health in a group of Iranian adolescents. A total of 1822 healthy adolescents (mean age = 13.97) from 25 high schools in Qazvin, Iran, completed a self‐reported survey at baseline and 6 months later. Structural equation modelling (SEM) was used to delineate the pathway from adolescents’ sleep hygiene knowledge, TPB constructs of their behavioural intentions and sleep hygiene behaviours and their sleep quality and self‐reported health. The SEM model demonstrated that although behavioural intention, coping planning and action planning predicted the sleep hygiene behaviours positively 6 months later with acceptable model fit [comparative fit index (CFI) = 0.936; Tucker–Lewis index (TLI) = 0.902; root mean square error of approximation (RMSEA) = 0.080; standardized root mean square residual (SRMR) = 0.044], sleep hygiene knowledge did not predict behavioural intentions significantly. Sleep hygiene behaviours were associated with sleep quality and psychiatric wellbeing. Thus, the TPB, combined with coping and action planning, is useful in understanding the sleep hygiene behaviours of adolescents. Health‐care providers may want to emphasize TPB constructs and coping and action planning to improve adolescents’ sleep hygiene behaviours, rather than rely solely upon increasing adolescents’ sleep hygiene knowledge.
Validity of spectral analysis based on heart rate variability from 1‐minute or less ECG recordings
Authors: Takahashi Naomi , Kuriyama Akira , Kanazawa Hoshinori , Takahashi Yoshimitsu , Nakayama Takeo
Published: June 2017

Background

To broaden the utility of heart rate variability (HRV) in clinical medicine and mass screening, results based on shorter electrocardiogram (ECG) recordings require validation with those based on standard 5‐minute recordings. We investigated the association between HRV variables obtained from 5‐minute ECGs with those obtained from ECGs shorter than 5 minutes.

Methods

Twenty‐two participants aged 20–69 years underwent 5‐minute resting ECG recordings in the supine position with natural breathing. Spectral analysis using MemCalc method was performed to calculate high‐frequency (HF, which required at least 10 seconds) and low‐frequency (LF, which required at least 30 seconds) components. Participants were not strictly preconditioned as in previous experimental studies in order to simulate a setting similar to that of a general health checkup. Associations of each variable between the 5‐minute ECG recordings and those for shorter recordings were examined by Pearson's correlation coefficients and Bland‐Altman plots.

Results

HF and LF components were log‐transformed based on their distributions. Correlation coefficients between 5‐minute data and shorter recordings in the supine position with natural breathing ranged from 0.80 to 0.91 (HF by 10‐second recording, 0.80; LF by 30‐second recording, 0.83, respectively). Bland‐Altman plots showed that gaps between the values from both methods slightly increased as the HF and LF component values increased.

Conclusions

Although slight proportional errors were possible, values from standard 5‐minute and shorter recordings in the supine position were strongly correlated. Our findings suggest that shorter ECG data without strict preconditioning can be reliably used for spectral analysis.
Binaural sensitivity in children who use bilateral cochlear implants
Authors: Ehlers Erica , Goupell Matthew J. , Zheng Yi , Godar Shelly P. , Litovsky Ruth Y.
Published: June 2017

Children who are deaf and receive bilateral cochlear implants (BiCIs) perform better on spatial hearing tasks using bilateral rather than unilateral inputs; however, they underperform relative to normal-hearing (NH) peers. This gap in performance is multi-factorial, including the inability of speech processors to reliably deliver binaural cues. Although much is known regarding binaural sensitivity of adults with BiCIs, less is known about how the development of binaural sensitivity in children with BiCIs compared to NH children. Sixteen children (ages 9–17 years) were tested using synchronized research processors. Interaural time differences and interaural level differences (ITDs and ILDs, respectively) were presented to pairs of pitch-matched electrodes. Stimuli were 300-ms, 100-pulses-per-second, constant-amplitude pulse trains. In the first and second experiments, discrimination of interaural cues (either ITDs or ILDs) was measured using a two-interval left/right task. In the third experiment, subjects reported the perceived intracranial position of ITDs and ILDs in a lateralization task. All children demonstrated sensitivity to ILDs, possibly due to monaural level cues. Children who were born deaf had weak or absent sensitivity to ITDs; in contrast, ITD sensitivity was noted in children with previous exposure to acoustic hearing. Therefore, factors such as auditory deprivation, in particular, lack of early exposure to consistent timing differences between the ears, may delay the maturation of binaural circuits and cause insensitivity to binaural differences.
Human echolocation in different situations and rooms
Authors: Schenkman Bo N.
Published: June 2017

People, especially when blind, use echolocation to detect obstacles, orient themselves, and get an awareness of their environment. I and coworkers have, with mostly psychophysical methods, studied perceptual aspects of how people accomplish echolocation. Echolocation with long canes while walking was possible but difficult. The effects of the spectral composition of the emitted sounds had no effects. Sound recordings in anechoic and conference rooms from non-walking, static situations, later presented in a laboratory showed a better performance in an ordinary room with reflections, than in an anechoic room. We also found that there was a higher performance with longer sounding sounds than for short clicks. Among the difficulties for the blind are how to avoid masking of sounds. A few blind are exceptionally high performing. An “information-surplus principle” has been proposed. Various information sources are used, but repetition pitch seems more important than loudness for echolocation. Among other sources, timbre may also provide information. There may exist a time gap, acoustic gaze, for how blind people use clicks. It is likely also that there are at least two processes taking place in the hearing system when listening for echoes, one attuned to short sounds and one to long sounds.
Osmoregulated Periplasmic Glucans
Authors: Bontemps-Gallo Sébastien , Bohin Jean-Pierre , Lacroix Jean-Marie
Published: June 2017

Among all the systems developed by enterobacteria to face osmotic stress, only osmoregulated periplasmic glucans (OPGs) were found to be modulated during osmotic fluxes. First detected in 1973 by E.P. Kennedy’s group in a study of phospholipid turnover in
Escherichia coli
, OPGs have been shown across alpha, beta, and gamma subdivisions of the proteobacteria. Discovery of OPG-like compounds in the epsilon subdivision strongly suggested that the presence of periplasmic glucans is essential for almost all proteobacteria. This article offers an overview of the different classes of OPGs. Then, the biosynthesis of OPGs and their regulation in
E. coli
and other species are discussed. Finally, the biological role of OPGs is developed. Beyond structural function, OPGs are involved in pathogenicity, in particular, by playing a role in signal transduction pathways. Recently, OPG synthesis proteins have been suggested to control cell division and growth rate.
Development of a Web Tool for Escherichia coli Subtyping Based on
fimH
Alleles
Authors: Roer Louise , Tchesnokova Veronika , Allesøe Rosa , Muradova Mariya , Chattopadhyay Sujay , Ahrenfeldt Johanne , Thomsen Martin C. F. , Lund Ole , Hansen Frank , Hammerum Anette M. , Sokurenko Evgeni , Hasman Henrik
Published: June 2017

The aim of this study was to construct a valid publicly available method for
in silico fimH
subtyping of
Escherichia coli
particularly suitable for differentiation of fine-resolution subgroups within clonal groups defined by standard multilocus sequence typing (MLST). FimTyper was constructed as a FASTA database containing all currently known
fimH
alleles. The software source code is publicly available at
https://bitbucket.org/genomicepidemiology/fimtyper
, the database is freely available at
https://bitbucket.org/genomicepidemiology/fimtyper_db
, and a service implementing the software is available at
https://cge.cbs.dtu.dk/services/FimTyper
. FimTyper was validated on three data sets: one containing Sanger sequences of
fimH
alleles of 42
E. coli
isolates generated prior to the current study (data set 1), one containing whole-genome sequence (WGS) data of 243 third-generation-cephalosporin-resistant
E. coli
isolates (data set 2), and one containing a randomly chosen subset of 40
E. coli
isolates from data set 2 that were subjected to conventional
fimH
subtyping (data set 3). The combination of the three data sets enabled an evaluation and comparison of FimTyper on both Sanger sequences and WGS data. FimTyper correctly predicted all 42
fimH
subtypes from the Sanger sequences from data set 1 and successfully analyzed all 243 draft genomes from data set 2. FimTyper subtyping of the Sanger sequences and WGS data from data set 3 were in complete agreement. Additionally,
fimH
subtyping was evaluated on a phylogenetic network of 122 sequence type 131 (ST131)
E. coli
isolates. There was perfect concordance between the typology and
fimH
-based subclones within ST131, with accurate identification of the pandemic multidrug-resistant clonal subgroup ST131-
H
30. FimTyper provides a standardized tool, as a rapid alternative to conventional
fimH
subtyping, highly suitable for surveillance and outbreak detection.
HDM2 Promotes NEDDylation of Hepatitis B Virus HBx To Enhance Its Stability and Function
Authors: Liu Ningning , Zhang Jinfang , Yang Xiaohai , Jiao Tong , Zhao Xin , Li Wenxia , Zhu Jianhua , Yang Pu , Jin Jianping , Peng Jirun , Li Zhiwei , Ye Xin
Published: June 2017

Hepatitis B virus (HBV)-encoded X protein (HBx) plays a critical role in HBV-related hepatocarcinoma development. In this study, we demonstrate that HBx is specifically modified by NEDD8. We found that E3 ligase HDM2 promotes NEDDylation of HBx to enhance HBx stability by preventing its ubiquitination-mediated degradation. Consistently, analysis of 160 hepatocellular carcinoma patient specimens indicated that the amount of HDM2 protein correlates with HBx protein level. We identified that HBx K91 and K95 as the key HBx NEDDylation sites and observed that the NEDDylation-deficient HBx has shorter half-life. We generated Huh7 cell lines which ectopically express wild-type and NEDDylation-deficient HBx and found that NEDDylation-deficient HBx showed less chromatin localization and less DDB1 binding. Consistently, the expression of HBx-regulated genes (IL-8, MMP9, and YAP) and HBV transcription (the activity of HBV enhancer and the amount of pgRNA transcribed from cccDNA) were significantly higher in cells expressing wild-type (WT) HBx than that in cells expressing mutant HBx. In addition, HBx-expressing cells proliferated faster than control and mutant HBx-expressing cells. We also showed that the ability of WT HBx-expressing cells to form tumors in nude mice was significantly higher than that of mutant HBx-expressing cells. In conclusion, we revealed that E3 ligase HDM2 promotes NEDDylation of HBx to enhance HBx stability and chromatin localization, which in turn favors HBx-dependent transcriptional regulation, cell proliferation, and HBV-driven tumor growth.

IMPORTANCE
Hepatitis B virus (HBV) HBx protein plays a critical role in viral replication and hepatocarcinogenesis. However, the regulation of HBx stability is not well understood. We found that HBx is modified by NEDD8 and that the HDM2 E3 ligase promotes HBx NEDDylation to enhance HBx stability by inhibiting its ubiquitination. We provide a new evidence to show the positive correlation between HDM2 and HBx in clinical hepatocellular carcinoma (HCC) samples. We also identified the major NEDDylation sites on HBx. Our studies indicate that the defective NEDDylation of HBx negatively affects its ability to activate the transcription of downstream genes and promote cell proliferation and tumor growth
in vivo
. Taken together, our findings reveal a novel posttranslational modification of HBx by HDM2 which regulates its stability, subcellular localization, and functions. These findings indicate that HDM2 is an important regulator on HBx and a potential diagnosis/therapeutic marker for HBV-associated HCC.
Evasion of the STING DNA-Sensing Pathway by VP11/12 of Herpes Simplex Virus 1
Authors: Deschamps Thibaut , Kalamvoki Maria
Published: June 2017

The
st
imulator of
in
terferon (IFN)
g
enes (STING) is a broad antimicrobial factor that restricts herpes simplex virus (HSV) by activating type I interferon and proinflammatory responses upon sensing of foreign DNA. UL46 is one of the most abundant tegument proteins of HSV-1, but a well-established function has yet to be found. We found that the HSV-1 UL46 protein interacts with and colocalizes with STING. A ΔUL46 virus displayed growth defects and activated innate immunity, but both effects were alleviated in STING knockdown cells. UL46 was also required for the inhibition of the 2′,3′-cyclic GMP-AMP (cGAMP)-dependent immune responses during infection. In cells expressing UL46, out of the context of the infection, innate immunity to a ΔICP0 virus was largely compromised, and that permitted ICP0-deficient mutants to replicate. The UL46-expressing cell lines also rescued the defects of the ΔUL46 virus and enhanced wild-type virus infection. The UL46-expressing cell lines did not activate interferon-stimulated gene (ISG) transcription following treatment with the noncanonical cyclic dinucleotide 2′,3′-cGAMP, suggesting that the STING pathway may be compromised. Indeed, we found that both proteins STING and IFI16 were eliminated in cells constitutively expressing UL46 and that the accumulation of their transcripts was blocked. Finally, we demonstrated that UL46 via its N terminus binds to STING and, via its C terminus, to TBK1. These interactions appear to modulate the functions of STING during HSV-1 infection. Taken together, our studies describe a novel function for one of the least-studied proteins of HSV, the tegument protein UL46, and that function involves the evasion of foreign DNA-sensing pathways.

IMPORTANCE
Herpes simplex virus 1 (HSV-1) afflicts 80% of the population worldwide, causing various diseases. After initial infection, the virus establishes latent reservoirs in sensory neurons and persists for life. Here we describe novel interactions between HSV-1 and the DNA sensor STING. We found that (i) HSV-1 tegument protein UL46 interacts with and colocalizes with STING; (ii) UL46 expressed out of the context of the infection blocks type I interferon triggered by STING stimuli, through the elimination of STING and of interferon-inducible protein 16 (IFI16); (iii) a ΔUL46 virus displayed growth defects, which were rescued in STING knockdown cells; (iv) the ΔUL46 virus failed to block innate immunity triggered by ligands of STING such as 2′,3′-cGAMP and also activated IFN-β and ISG expression; and (v) UL46 binds to both STING and TBK1 through different domains. We conclude that UL46 counteracts the actions of STING during HSV-1 infection.
Guanylate-Binding Protein 1 Inhibits Nuclear Delivery of Kaposi's Sarcoma-Associated Herpesvirus Virions by Disrupting Formation of Actin Filament
Authors: Zou Zhe , Meng Zhihua , Ma Chao , Liang Deguang , Sun Rui , Lan Ke
Published: June 2017

Kaposi's sarcoma-associated herpesvirus (KSHV) is a typical gammaherpesvirus that establishes persistent lifelong infection in host cells. In order to establish successful infection, KSHV has evolved numerous immune evasion strategies to bypass or hijack the host immune system. However, host cells still produce immune cytokines abundantly during primary KSHV infection. Whether the immune effectors produced are able to inhibit viral infection and how KSHV successfully conquers these immune effectors remain largely unknown. The guanylate-binding protein 1 (GBP1) gene is an interferon-stimulated gene and exerts antiviral functions on several RNA viruses; however, its function in DNA virus infection is less well understood. In this study, we found that KSHV infection increases both the transcriptional and protein levels of GBP1 at the early stage of primary infection by activating the NF-κB pathway. The overexpression of GBP1 significantly inhibited KSHV infection, while the knockdown of GBP1 promoted KSHV infection. The GTPase activity and dimerization of GBP1 were demonstrated to be responsible for its anti-KSHV activity. Furthermore, we found that GBP1 inhibited the nuclear delivery of KSHV virions by disrupting the formation of actin filaments. Finally, we demonstrated that replication and transcription activator (RTA) promotes the degradation of GBP1 through a proteasome pathway. Taken together, these results provide a new understanding of the antiviral mechanism of GBP1, which possesses potent anti-KSHV activity, and suggest the critical role of RTA in the evasion of the innate immune response during primary infection by KSHV.

IMPORTANCE
GBP1 can be induced by various cytokines and exerts antiviral activities against several RNA viruses. Our study demonstrated that GBP1 can exert anti-KSHV function by inhibiting the nuclear delivery of KSHV virions via the disruption of actin filaments. Moreover, we found that KSHV RTA can promote the degradation of GBP1 through a proteasome-mediated pathway. Taken together, our results elucidate a novel mechanism of GBP1 anti-KSHV activity and emphasize the critical role of RTA in KSHV evasion of the host immune system during primary infection.
Gender Parity Trends for Invited Speakers at Four Prominent Virology Conference Series
Authors: Kalejta Robert F. , Palmenberg Ann C.
Published: June 2017

Scientific conferences are most beneficial to participants when they showcase significant new experimental developments, accurately summarize the current state of the field, and provide strong opportunities for collaborative networking. A top-notch slate of invited speakers, assembled by conference organizers or committees, is key to achieving these goals. The perceived underrepresentation of female speakers at prominent scientific meetings is currently a popular topic for discussion, but one that often lacks supportive data. We compiled the full rosters of invited speakers over the last 35 years for four prominent international virology conferences, the American Society for Virology Annual Meeting (ASV), the International Herpesvirus Workshop (IHW), the Positive-Strand RNA Virus Symposium (PSR), and the Gordon Research Conference on Viruses & Cells (GRC). The rosters were cross-indexed by unique names, gender, year, and repeat invitations. When plotted as gender-dependent trends over time, all four conferences showed a clear proclivity for male-dominated invited speaker lists. Encouragingly, shifts toward parity are emerging within all units, but at different rates. Not surprisingly, both selection of a larger percentage of first-time participants and the presence of a woman on the speaker selection committee correlated with improved parity. Session chair information was also collected for the IHW and GRC. These visible positions also displayed a strong male dominance over time that is eroding slowly. We offer our personal interpretation of these data to aid future organizers achieve improved equity among the limited number of available positions for session moderators and invited speakers.

IMPORTANCE
Politicians and media members have a tendency to cite anecdotes as conclusions without any supporting data. This happens so frequently now, that a name for it has emerged: fake news. Good science proceeds otherwise. The underrepresentation of women as invited speakers at international scientific conferences exemplifies a present-day discussion topic usually occurring without facts to support or refute the arguments. We now provide records profiling four prominent virology conferences over the years 1982 to 2017 with the intention that the trends and accompanying analyses of the gender parity of invited speakers may allow the ongoing discussions to be informed.
Newcastle Disease Virus Establishes Persistent Infection in Tumor Cells
In Vitro
: Contribution of the Cleavage Site of Fusion Protein and Second Sialic Acid Binding Site of Hemagglutinin-Neuraminidase
Authors: Rangaswamy Udaya S. , Wang Weijia , Cheng Xing , McTamney Patrick , Carroll Danielle , Jin Hong
Published: June 2017

Newcastle disease virus (NDV) is an oncolytic virus being developed for the treatment of cancer. Following infection of a human ovarian cancer cell line (OVCAR3) with a recombinant low-pathogenic NDV, persistent infection was established in a subset of tumor cells. Persistently infected (PI) cells exhibited resistance to superinfection with NDV and established an antiviral state, as demonstrated by upregulation of interferon and interferon-induced genes such as myxoma resistance gene 1 (Mx1) and retinoic acid-inducing gene-I (RIG-I). Viruses released from PI cells induced higher cell-to-cell fusion than the parental virus following infection in two tumor cell lines tested, HT1080 and HeLa, and remained attenuated in chickens. Two mutations, one in the fusion (F) protein cleavage site, F117S (F
117S
), and another in hemagglutinin-neuraminidase (HN), G169R (HN
169R
), located in the second sialic acid binding region, were responsible for the hyperfusogenic phenotype. F
117S
improves F protein cleavage efficiency, facilitating cell-to-cell fusion, while HN
169R
possesses a multifaceted role in contributing to higher fusion, reduced receptor binding, and lower neuraminidase activity, which together result in increased fusion and reduced viral replication. Thus, establishment of persistent infection
in vitro
involves viral genetic changes that facilitate efficient viral spread from cell to cell as a potential mechanism to escape host antiviral responses. The results of our study also demonstrate a critical role in the viral life cycle for the second receptor binding region of the HN protein, which is conserved in several paramyxoviruses.

IMPORTANCE
Oncolytic Newcastle disease virus (NDV) could establish persistent infection in a tumor cell line, resulting in a steady antiviral state reflected by constitutively expressed interferon. Viruses isolated from persistently infected cells are highly fusogenic, and this phenotype has been mapped to two mutations, one each in the fusion (F) and hemagglutinin-neuraminidase (HN) proteins. The F
117S
mutation in the F protein cleavage site improved F protein cleavage efficiency while the HN
169R
mutation located at the second receptor binding site of the HN protein contributed to a complex phenotype consisting of a modest increase in fusion and cell killing, lower neuraminidase activity, and reduced viral growth. This study highlights the intricate nature of these two mutations in the glycoproteins of NDV in the establishment of persistent infection. The data also shed light on the critical balance between the F and HN proteins required for efficient NDV infection and their role in avian pathogenicity.
Psyllids, It’s What’s on the Inside That Counts: Community Cross Talk Facilitates Prophage Interactions
Authors: Hansen Allison K. , Skidmore Isabel H.
Published: June 2017

Despite the availability of massive microbial community data sets (e.g., metagenomes), there is still a lack of knowledge on what molecular mechanisms facilitate cross talk between microbes and prophage within a community context.
Malignant hypertension as a rare cause of thrombotic microangiopathy
Authors: Thind Guramrinder , Kailasam Karthik
Published: June 2017

Malignant hypertension can occasionally be associated with microangiopathic haemolytic anaemia. A 38-year-old male presented with nausea, vomiting, loss of appetite and oliguria for 2 weeks. He was diagnosed with hypertensive emergency with cardiac and renal dysfunction. Interestingly, further workup was diagnostic for the presence of thrombotic microangiopathy (TMA): haemoglobin =12.7 g/dL, indirect bilirubin =2.0 mg/dL, haptoglobin ≤6 mg/dL, platelet count =121 000/μL and schistocytes on peripheral smear. At the outset, the cause of TMA was unclear. Patient denied having diarrhoea, making haemolytic uremic syndrome less likely. A normal ADAMTS13 activity test ruled out thrombotic thrombocytopaenic purpura. Malignant hypertension induced TMA was highest on the differential and plasma exchange was deferred. Renal biopsy revealed features of TMA and malignant nephrosclerosis. Patient eventually became dialysis dependent. Aggressive blood pressure control was obtained with multiple medications.
Growth of meibomian gland tissue after intraductal meibomian gland probing in patients with obstructive meibomian gland dysfunction
Authors: Maskin Steven L , Testa Whitney R
Published: June 2017

Purpose

To investigate the impact of meibomian gland probing (MGP) on meibomian gland (MG) area from the upper lids of patients with obstructive meibomian gland dysfunction (o-MGD).

Methods

Retrospective study comparing pre-MGP/post-MGP non-contact infrared meibography results in patients with o-MGD, viewing signs of MG growth within total measurement field.

Results

Post-MGP meibography of 34 lids (19 patients, ≥4.5 to ≤12 months’ follow-up) showed 41.2% with MG growth. Ten lids had meibographies suitable for analysis, showing significant collective (116 glands) increase in mean individual glandular area (MIGA) of 4.87% (p=0.0145). Four of 10 lids independently showed significant increase in MIGA, ranging from 10.70% to 21.13% (p<0.0001, p=0.0277, p=0.0292, p=0.0345), while six did not.

At >12 and <25 months’ follow-up, 16 lids (9 additional patients) had follow-up showing 25% with signs of MG growth. Analysis of three lids showed a significant collective (33 glands) increase in MIGA of 11.19% (p=0.0004). Two of three lids independently showed significant increase in MIGA of 13.73% and 20.00% (p=0.0097, p=0.0001). Collectively, for all 13 analysed lids (149 glands), there was a significant increase of 6.38% in total glandular area (p=0.0447) and a significant increase of 6.23% in MIGA (p=0.0003).

Conclusion

MGP was associated with increased MG tissue area and growth of atrophied MGs as viewed on meibography. MGP provides unequivocal physical proof of a patent meibum outflow tract through the natural orifice, and may promote glandular growth in part by direct mechanical establishment of a patent duct/orifice system.
Resisting the urge to prescribe vancomycin
Authors: Magalhães‐Sant'Ana Manuel
Published: June 2017

THIS series gives readers the opportunity to consider and contribute to discussion of some of the ethical dilemmas that can arise in veterinary practice. Each month, a case scenario is presented, followed by discussion of some of the issues involved.

In addition, a possible way forward is suggested; however, there is rarely a cut‐and‐dried answer in such cases, and readers may wish to suggest an alternative approach. This month's dilemma, ‘Resisting the urge to prescribe vancomycin’, was submitted and is discussed by Manuel Magalhães‐Sant'Ana. Readers with comments to contribute are invited to send them as soon as possible, so that they can be considered for publication in the next issue.

The series is being coordinated by Steven McCulloch, a practising vet with a PhD in the ethics of veterinary policy. It aims to provide a framework that will help practitioners find solutions when facing similar dilemmas.
Approach to the cryptorchid horse
Authors: Pollock Patrick
Published: June 2017

Cryptorchidism (‘hidden testis’) is the failure of one or both testes to descend from the developmental position close to the kidney to the normal adult position in the scrotum. The term cryptorchid is used to describe the non‐descended testis and, by extension, affected horses, although a number of colloquial terms are also used for the condition, including rig, ridgling and original. Cryptorchidism is thought to be an inherited condition and is the most common, non‐lethal developmental abnormality in horses. Although it has been reported in a variety of different breeds, those most commonly affected include percherons, saddlebreds, quarter horses, native‐breed ponies and crossbred horses; thoroughbreds have the lowest prevalence. This article gives an overview of the anatomy of the testes and describes the challenges involved in diagnosing the condition and the techniques that are available for treatment.
Analysis of neointima development in flow diverters using optical coherence tomography imaging
Authors: Matsuda Yoshikazu , Chung Joonho , Lopes Demetrius K
Published: June 2017

Background

Flow diverters are used for the treatment of intracranial aneurysms. Surface modification may decrease the thrombogenicity of flow diverters but the details are unknown. Optical coherence tomography (OCT) is an intravascular imaging test with high resolution which identifies neointimal growth over stents. We compared the development of neointima in a flow diverter and stents with and without surface modification in a swine model.

Methods

In this study we implanted four devices (two in each carotid artery) in four pigs. The devices used were the Pipeline Flex embolization device (PED Flex, n=6), PED with Shield technology (PED Shield, n=6), and Solitaire AB (n=4). Serial carotid angiographic and OCT images were obtained on days 0, 7, 14, and 21. The data analyzed included: neointimal area (lumen area − stent area), neointimal ratio ([lumen area − stent area]/stent area), and the neointimal thickness ratio (minimum neointimal thickness/maximum neointimal thickness).

Results

There was no significant difference in where neointima formation was initiated in relation to the implanted device (distal vs middle vs proximal). The PED Shield had a trend towards earlier endothelial formation at day 7. By day 21 the neointimal ratio was significantly higher for the PED Flex and PED Shield devices than for Solitaire (p<0.05 and p<0.01, respectively). The neointimal thickness ratio was significantly higher with PED Shield than with PED Flex and Solitaire (p<0.05 and p<0.01, respectively).

Conclusions

OCT enabled us to follow and compare in vivo the development of neointima over implants. PED Shield showed a similar neointimal volume to PED Flex and more concentric neointima.