bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mutations in the voltage‐sensing domain affect the alternative ion permeation pathway in the TRPM3 channel
Authors: Held Katharina , Gruss Fabian , Aloi Vincenzo Davide , Janssens Annelies , Ulens Chris , Voets Thomas , Vriens Joris 
Published: March 2018

Key points
Mutagenesis at positively charged amino acids (arginines and lysines) (R1–R4) in the voltage‐sensor domain (transmembrane segment (S) 4) of voltage‐gated Na+, K+ and Ca2+ channels can lead to an alternative ion permeation pathway distinct from the central pore.
Recently, a non‐canonical ion permeation pathway was described in TRPM3, a member of the transient receptor potential (TRP) superfamily. The non‐canonical pore exists in the native TRPM3 channel and can be activated by co‐stimulation of the endogenous agonist pregnenolone sulphate and the antifungal drug clotrimazole or by stimulation of the synthetic agonist CIM0216.
Alignment of the voltage sensor of Shaker K+ channels with the entire TRPM3 sequence revealed the highest degree of similarity in the putative S4 region of TRPM3, and suggested that only one single gating charge arginine (R2) in the putative S4 region is conserved.
Mutagenesis studies in the voltage‐sensing domain of TRPM3 revealed several residues in the voltage sensor (S4) as well as in S1 and S3 that are crucial for the occurrence of the non‐canonical inward currents.
In conclusion, this study provides evidence for the involvement of the voltage‐sensing domain of TRPM3 in the formation of an alternative ion permeation pathway.
AbstractTransient receptor potential (TRP) channels are cationic channels involved in a broad array of functions, including homeostasis, motility and sensory functions. TRP channel subunits consist of six transmembrane segments (S1–S6), and form tetrameric channels with a central pore formed by the region encompassing S5 and S6. Recently, evidence was provided for the existence of an alternative ion permeation pathway in TRPM3, which allows large inward currents upon hyperpolarization independently of the central pore. However, very little knowledge is available concerning the localization of this alternative pathway in the native TRPM3 channel protein. Guided by sequence homology with Shaker K+ channels, in which mutations in S4 can create an analogous ‘omega’ pore, we performed site‐directed mutagenesis studies and patch clamp experiments to identify amino acid residues involved in the formation of the non‐canonical pore in TRPM3. Based on our results, we pinpoint four residues in S4 (W982, R985, D988 and G991) as crucial determinants of the properties of the alternative ion permeation pathway.
Protein Acetylation Mediated by YfiQ and CobB Is Involved in the Virulence and Stress Response of Yersinia pestis
Authors: Liu Wanbing , Tan Yafang , Cao Shiyang , Zhao Haihong , Fang Haihong , Yang Xiaoyan , Wang Tong , Zhou Yazhou , Yan Yanfeng , Han Yanping , Song Yajun , Bi Yujing , Wang Xiaoyi , Yang Ruifu , Du Zongmin 
Published: April 2018

ABSTRACT

Recent studies revealed that acetylation is a widely used protein modification in prokaryotic organisms. The major protein acetylation acetyltransferase YfiQ and the sirtuin-like deacetylase CobB have been found to be involved in basic physiological processes, such as primary metabolism, chemotaxis, and stress responses, in
Escherichia coli
and
Salmonella
. However, little is known about protein acetylation modifications in
Yersinia pestis
, a lethal pathogen responsible for millions of human deaths in three worldwide pandemics. Here we found that
Yp_0659
and
Yp_1760
of
Y. pestis
encode the major protein acetylation acetyltransferase YfiQ and the sirtuin-like deacetylase CobB, respectively, which can acetylate and deacetylate PhoP enzymatically
in vitro
. Protein acetylation impairment in
cobB
and
yfiQ
mutants greatly decreased bacterial tolerance to cold, hot, high-salt, and acidic environments. Our comparative transcriptomic data revealed that the strongly decreased tolerance to stress stimuli was probably related to downregulation of the genes encoding the heat shock proteins (HtpG, HslV, HslR, and IbpA), cold shock proteins (CspC and CspA1), and acid resistance proteins (HdeB and AdiA). We found that the reversible acetylation mediated by CobB and YfiQ conferred attenuation of virulence, probably partially due to the decreased expression of the
psaABCDEF
operon, which encodes Psa fimbriae that play a key role in virulence of
Y. pestis
. This is the first report, to our knowledge, on the roles of protein acetylation modification in stress responses, biofilm formation, and virulence of
Y. pestis
.
Feline Herpesvirus 1 US3 Blocks the Type I Interferon Signal Pathway by Targeting Interferon Regulatory Factor 3 Dimerization in a Kinase-Independent Manner
Authors: Tian Jin , Liu Yongxiang , Liu Xiaoxiao , Sun Xue , Zhang Jikai , Qu Liandong 
Published: April 2018

ABSTRACT

As a prevalent agent in cats, feline herpesvirus 1 (FHV-1) infection contributes to feline respiratory disease and acute and chronic conjunctivitis. FHV-1 can successfully evade the host innate immune response and persist for the lifetime of the cat. Several mechanisms of immune evasion by human herpesviruses have been elucidated, but the mechanism of immune evasion by FHV-1 remains unknown. In this study, we screened for FHV-1 open reading frames (ORFs) responsible for inhibiting the type I interferon (IFN) pathway with an IFN-β promoter reporter and analysis of IFN-β mRNA levels in HEK 293T cells and the Crandell-Reese feline kidney (CRFK) cell line, and we identified the Ser/Thr kinase US3 as the most powerful inhibitor. Furthermore, we found that the anti-IFN activity of US3 depended on its N terminus (amino acids 1 to 75) and was independent of its kinase activity. Mechanistically, the ectopic expression of US3 selectively inhibited IFN regulatory factor 3 (IRF3) promoter activation. Furthermore, US3 bound to the IRF association domain (IAD) of IRF3 and prevented IRF3 dimerization. Finally, US3-deleted recombinant FHV-1 and US3-repaired recombinant FHV-1 (rFHV-dUS3 and rFHV-rUS3, respectively) were constructed. Compared with wild-type FHV-1 and rFHV-rUS3, infection with rFHV-dUS3 induced large amounts of IFN-β
in vitro
and
in vivo
. More importantly, US3 deletion significantly attenuated virulence, reduced virus shedding, and blocked the invasion of trigeminal ganglia. These results indicate that FHV-1 US3 efficiently inhibits IFN induction by using a novel immune evasion mechanism and that FHV-1 US3 is a potential regulator of neurovirulence.

IMPORTANCE
Despite widespread vaccination, the prevalence of FHV-1 remains high, suggesting that it can successfully evade the host innate immune response and infect cats. In this study, we screened viral proteins for inhibiting the IFN pathway and identified the Ser/Thr kinase US3 as the most powerful inhibitor. In contrast to other members of the alphaherpesviruses, FHV-1 US3 blocked the host type I IFN pathway in a kinase-independent manner and via binding to the IRF3 IAD and preventing IRF3 dimerization. More importantly, the depletion of US3 attenuated the anti-IFN activity of FHV-1 and prevented efficient viral replication
in vitro
and
in vivo
. Also, US3 deletion significantly attenuated virulence and blocked the invasion of trigeminal ganglia. We believe that these findings not only will help us to better understand the mechanism of how FHV-1 manipulates the host IFN response but also highlight the potential role of US3 in the establishment of latent infection
in vivo
.
Identification of the RNA Pseudoknot within the 3′ End of the Porcine Reproductive and Respiratory Syndrome Virus Genome as a Pathogen-Associated Molecular Pattern To Activate Antiviral Signaling via RIG-I and Toll-Like Receptor 3
Authors: Xie Sha , Chen Xin-xin , Qiao Songlin , Li Rui , Sun Yangang , Xia Shuangfei , Wang Lin-Jian , Luo Xuegang , Deng Ruiguang , Zhou En-Min , Zhang Gai-Ping 
Published: April 2018

ABSTRACT

Once infected by viruses, cells can detect pathogen-associated molecular patterns (PAMPs) on viral nucleic acid by host pattern recognition receptors (PRRs) to initiate the antiviral response. Porcine reproductive and respiratory syndrome virus (PRRSV) is the causative agent of porcine reproductive and respiratory syndrome (PRRS), characterized by reproductive failure in sows and respiratory diseases in pigs of different ages. To date, the sensing mechanism of PRRSV has not been elucidated. Here, we reported that the pseudoknot region residing in the 3′ untranslated regions (UTR) of the PRRSV genome, which has been proposed to regulate RNA synthesis and virus replication, was sensed as nonself by retinoic acid-inducible gene I (RIG-I) and Toll-like receptor 3 (TLR3) and strongly induced type I interferons (IFNs) and interferon-stimulated genes (ISGs) in porcine alveolar macrophages (PAMs). The interaction between the two stem-loops inside the pseudoknot structure was sufficient for IFN induction, since disruption of the pseudoknot interaction powerfully dampened the IFN induction. Furthermore, transfection of the 3′ UTR pseudoknot transcripts in PAMs inhibited PRRSV replication
in vitro
. Importantly, the predicted similar structures of other arterivirus members, including equine arteritis virus (EAV), lactate dehydrogenase-elevating virus (LDV), and simian hemorrhagic fever virus (SHFV), also displayed strong IFN induction activities. Together, in this work we identified an innate recognition mechanism by which the PRRSV 3′ UTR pseudoknot region served as PAMPs of arteriviruses and activated innate immune signaling to produce IFNs that inhibit virus replication. All of these results provide novel insights into innate immune recognition during virus infection.

IMPORTANCE
PRRS is the most common viral disease in the pork industry. It is caused by PRRSV, a positive single-stranded RNA virus, whose infection often leads to persistent infection. To date, it is not yet clear how PRRSV is recognized by the host and what is the exact mechanism of IFN induction. Here, we investigated the nature of PAMPs on PRRSV and the associated PRRs. We found that the 3′ UTR pseudoknot region of PRRSV, which has been proposed to regulate viral RNA synthesis, could act as PAMPs recognized by RIG-I and TLR3 to induce type I IFN production to suppress PRRSV infection. This report is the first detailed description of pattern recognition for PRRSV, which is important in understanding the antiviral response of arteriviruses, especially PRRSV, and extends our knowledge on virus recognition.
Clinical and Pathological Characteristics of KEAP1- and NFE2L2-Mutated Non–Small Cell Lung Carcinoma (NSCLC)
Authors: Frank Rieke , Scheffler Matthias , Merkelbach-Bruse Sabine , Ihle Michaela A. , Kron Anna , Rauer Michael , Ueckeroth Frank , König Katharina , Michels Sebastian , Fischer Rieke , Eisert Anna , Fassunke Jana , Heydt Carina , Serke Monika , Ko Yon-Dschun , Gerigk Ulrich , Geist Thomas , Kaminsky Britta , Heukamp Lukas C. , Clement-Ziza Mathieu , Büttner Reinhard , Wolf Jürgen 
Published: April 2018

Abstract
Purpose: KEAP1 and NFE2L2 mutations are associated with impaired prognosis in a variety of cancers and with squamous cell carcinoma formation in non–small cell lung cancer (NSCLC). However, little is known about frequency, histology dependence, molecular and clinical presentation as well as response to systemic treatment in NSCLC.
Experimental Design: Tumor tissue of 1,391 patients with NSCLC was analyzed using next-generation sequencing (NGS). Clinical and pathologic characteristics, survival, and treatment outcome of patients with KEAP1 or NFE2L2 mutations were assessed.
Results: KEAP1 mutations occurred with a frequency of 11.3% (n = 157) and NFE2L2 mutations with a frequency of 3.5% (n = 49) in NSCLC patients. In the vast majority of patients, both mutations did not occur simultaneously. KEAP1 mutations were found mainly in adenocarcinoma (AD; 72%), while NFE2L2 mutations were more common in squamous cell carcinoma (LSCC; 59%). KEAP1 mutations were spread over the whole protein, whereas NFE2L2 mutations were clustered in specific hotspot regions. In over 80% of the patients both mutations co-occurred with other cancer-related mutations, among them also targetable aberrations like activating EGFR mutations or MET amplification. Both patient groups showed different patterns of metastases, stage distribution and performance state. No patient with KEAP1 mutation had a response on systemic treatment in first-, second-, or third-line setting. Of NFE2L2-mutated patients, none responded to second- or third-line therapy.
Conclusions: KEAP1- and NFE2L2-mutated NSCLC patients represent a highly heterogeneous patient cohort. Both are associated with different histologies and usually are found together with other cancer-related, partly targetable, genetic aberrations. In addition, both markers seem to be predictive for chemotherapy resistance. Clin Cancer Res; 24(13); 3087–96. ©2018 AACR.
Genetic Analysis of 779 Advanced Differentiated and Anaplastic Thyroid Cancers
Authors: Pozdeyev Nikita , Gay Laurie M. , Sokol Ethan S. , Hartmaier Ryan , Deaver Kelsi E. , Davis Stephanie , French Jena D. , Borre Pierre Vanden , LaBarbera Daniel V. , Tan Aik-Choon , Schweppe Rebecca E. , Fishbein Lauren , Ross Jeffrey S. , Haugen Bryan R. , Bowles Daniel W. 
Published: April 2018

Abstract
Purpose: To define the genetic landscape of advanced differentiated and anaplastic thyroid cancer (ATC) and identify genetic alterations of potential diagnostic, prognostic, and therapeutic significance.
Experimental Design: The genetic profiles of 583 advanced differentiated and 196 ATCs generated with targeted next-generation sequencing cancer-associated gene panels MSK-IMPACT and FoundationOne were analyzed.
Results: ATC had more genetic alterations per tumor, and pediatric papillary thyroid cancer had fewer genetic alterations per tumor when compared with other thyroid cancer types. DNA mismatch repair deficit and activity of APOBEC cytidine deaminases were identified as mechanisms associated with high mutational burden in a subset of differentiated thyroid cancers and ATCs. Copy number losses and mutations of CDKN2A and CDKN2B, amplification of CCNE1, amplification of receptor tyrosine kinase genes KDR, KIT, and PDGFRA, amplification of immune evasion genes CD274, PDCD1LG2, and JAK2, and activating point mutations in small GTPase RAC1 were associated with ATC. An association of KDR, KIT, and PDGFRA amplification with the sensitivity of thyroid cancer cells to lenvatinib was shown in vitro. Three genetically distinct types of ATCs are proposed.
Conclusions: This large-scale analysis describes genetic alterations in a cohort of thyroid cancers enriched in advanced cases. Many novel genetic events previously not seen in thyroid cancer were found. Genetic alterations associated with anaplastic transformation were identified. An updated schematic of thyroid cancer genetic evolution is proposed. Clin Cancer Res; 24(13); 3059–68. ©2018 AACR.
TTR (Transthyretin) Stabilizers Are Associated With Improved Survival in Patients With TTR Cardiac Amyloidosis
Authors: Rosenblum Hannah , Castano Adam , Alvarez Julissa , Goldsmith Jeff , Helmke Stephen , Maurer Mathew S. 
Published: April 2018

Background:
TTR (transthyretin) cardiac amyloidosis is caused by dissociation of TTR into monomers, which misassemble into amyloid fibrils. TTR stabilizers act at the dimer–dimer interface to prevent dissociation. We investigated differences in survival among patients with TTR cardiac amyloidosis on stabilizer medications compared with those not on stabilizers.

Methods AND RESULTS:

A retrospective study of patients with TTR cardiac amyloidosis presenting to a single center was conducted. Baseline characteristics were compared between those treated with stabilizers and those not treated with stabilizers. Cox proportional hazards modeling assessed for univariate predictors of the composite outcome of death or orthotopic heart transplant (OHT). Multivariable Cox proportional hazards assessed whether stabilizer treatment was independently associated with improved death or OHT after controlling for significant univariate predictors. One hundred twenty patients (mean age, 75±8, 88% male) were included: 29 patients who received stabilizers and 91 patients who did not. Stabilizer use was associated with a lower risk of the combined end point of death or OHT (hazard ratio, 0.32; 95% confidence interval, 0.18–0.58;
P
<0.0001). Subjects treated with stabilizers were more likely to be of White race (93% versus 55%;
P
<0.001), classified as New York Heart Association classes I and II (79% versus 38%;
P
=0.002), less likely to have a mutation (10% versus 36%;
P
=0.010), have lower troponin I (median 0.06 versus 0.12 ng/mL;
P
=0.002), and higher left ventricular ejection fraction (49% versus 40%;
P
=0.011), suggesting earlier stage of disease. In multivariable Cox analysis, the association between stabilizer and death or OHT persisted when adjusted for all noncollinear univariate predictors with
P
<0.05 (hazard ratio, 0.37; 95% confidence interval, 0.19–0.75;
P
=0.003).

Conclusions:
TTR stabilizers are associated with decreased death and OHT in TTR cardiac amyloidosis. These results need to be confirmed by ongoing randomized clinical trials.
Rashbam’s Approach to Targum Onqelos in His Commentary to the Torah
Authors: Jacobs Jonathan 
Published: April 2018

Abstract
The Targum of Onqelos was the principal pentateuchal targum on which medieval Jews in northern France relied. When Rashbam referenced Onqelos, he used a consistent format composed of four elements. His use of the targum is varied: in most instances, his purpose was lexical. In about one-third of cases, his purpose was exegetical. In three cases, Rashbam cited the targum in order to consider its precise intention (while finding fault with the conclusion reached by Rashi in that regard), an indication of the great importance that Rashbam ascribed to the targum. Nevertheless, despite the high esteem in which he held the targum, Rashbam occasionally allowed himself to disagree with it in his quest to uncover the simplest sense of Scripture. Targum Onqelos is the work most often quoted in Rashbam’s commentary to the Torah, and his use of it demonstrates extensive knowledge of this targum to the Torah.
Erasmus on Dogs and Baths and Other Odious Comparisons
Authors: Eden Kathy 
Published: March 2018

Abstract
Fully aware of an antipathy to comparisons that looks back not only to ancient philosophy and law but to the early modern schoolroom, Erasmus nevertheless puts his full prestige behind the strategy so foundational to the rhetorical theory of Plato, Cicero, Quintilian and Aphthonius. This essay examines the key role of comparison in the form of similitudo, parabola or collatio, and imago in Erasmus’ educational reform as represented by his De copia, De ratione studii, and De conscribendis epistolis, as well as in his own literary production, especially his Adages.
Sea-Effect Precipitation over the Shandong Peninsula, Northern China
Authors: Bao Baoleerqimuge , Ren Guoyu 
Published: April 2018

AbstractSea-effect precipitation (SEP) over the Shandong Peninsula is a unique climatological phenomenon in mainland China, and it exerts a considerable impact on the southern shore of the Bohai Sea. From observed data from 123 stations for the period 1962–2012, the characteristics of cold-season (November–February) SEP in this area were analyzed. Results showed that SEP occurred throughout the late autumn and winter. In all, 1173 SEP days were identified during the 51 years, of which snow days accounted for 73.7% and rain and snow–rain days accounted for 16.1% and 10.1%, respectively. December had the largest number of SEP snow days, followed by January and November. November was the most productive month in terms of SEP rain and snow–rain days. Intense SEP snowfall mainly affected the inland hill area of the peninsula, whereas light SEP snowfall reached farther inland. SEP rainfall shared a similar pattern with snowfall. The SEP frequency showed a significant interannual variability and a nonsignificant upward trend over the period analyzed. SEP was most likely to occur when the temperature difference between sea surface and 850 hPa over the Bohai Sea was above 10°C, indicating a dominant influence of low-level cold-air advection over the sea on the generation and development of the weather phenomenon. A significant negative correlation was also found between the area of sea ice in the Bohai Sea and intense SEP snowfall, indicating that sea ice extent had an important effect on SEP variability over the peninsula. In the case of extremely intense SEP events, a deeper East Asian trough at the 500-hPa level developed over the southwest of the study area and temperature and geopotential height contours were orthogonal to each other, indicating strong geostrophic cold-air advection over the Bohai Sea and the Shandong Peninsula. The extremely intense SEP events were also characterized by anomalous low temperature and high relative humidity in the lower troposphere, which contributed to greater gravitational instability in the study area.
The Evidence Base for the Maternal, Infant, and Early Childhood Home Visiting Program Constructs
Authors: Campos Sofia , Kapp Julie M. , Simoes Eduardo J. 
Published: April 2018

Objectives: The federal Maternal, Infant, and Early Childhood Home Visiting (MIECHV) program requires grantees to demonstrate program improvement as a condition of funding. The MIECHV program monitors grantee progress in federally mandated conceptual areas (ie, benchmarks) that are further subcategorized into related sub-areas or constructs (eg, breastfeeding). Each construct has an associated performance measure that helps MIECHV collect data on program implementation and performance. In 2016, MIECHV modified the constructs and associated performance measures required of grantees. Our objective was to identify whether the constructs were supported by the home visiting literature. Methods: We conducted an evaluation of one of the MIECHV program’s benchmarks (Benchmark 1: Maternal and Newborn Health) for alignment of the Benchmark 1 constructs (preterm birth, breastfeeding, depression screening, well-child visit, postpartum care, and tobacco cessation referrals) with home visiting evidence. In March 2016, we searched the Home Visiting Evidence of Effectiveness database for all publicly available articles on studies conducted in the United States to determine how well the study findings aligned with the MIECHV program constructs. Results: Of 59 articles reviewed, only 3 of the 6 MIECHV constructs—preterm birth, breastfeeding, and well-child visits—were supported by home visiting evidence. Conclusions: This evaluation highlights a limited evidence base for the MIECHV Benchmark 1 constructs and a need to clarify other criteria, beyond evidence, used to choose constructs and associated performance measures. One implication of not having evidence-based performance measures is a lack of confidence that the program will drive positive outcomes. If performance measures are not evidence based, it is difficult to attribute positive outcomes to the home visiting services.