bluuebunny/crossref_metadata_2025_split huggingface dataset:
In Situ
Tagged nsp15 Reveals Interactions with Coronavirus Replication/Transcription Complex-Associated Proteins
Authors: Athmer Jeremiah , Fehr Anthony R. , Grunewald Matthew , Smith Everett Clinton , Denison Mark R. , Perlman Stanley 
Published: February 2017

ABSTRACT

Coronavirus (CoV) replication and transcription are carried out in close proximity to restructured endoplasmic reticulum (ER) membranes in replication/transcription complexes (RTC). Many of the CoV nonstructural proteins (nsps) are required for RTC function; however, not all of their functions are known. nsp15 contains an endoribonuclease domain that is conserved in the CoV family. While the enzymatic activity and crystal structure of nsp15 are well defined, its role in replication remains elusive. nsp15 localizes to sites of RNA replication, but whether it acts independently or requires additional interactions for its function remains unknown. To begin to address these questions, we created an
in situ
tagged form of nsp15 using the prototypic CoV, mouse hepatitis virus (MHV). In MHV, nsp15 contains the genomic RNA packaging signal (P/S), a 95-bp RNA stem-loop structure that is not required for viral replication or nsp15 function. Utilizing this knowledge, we constructed an internal hemagglutinin (HA) tag that replaced the P/S. We found that nsp15-HA was localized to discrete perinuclear puncta and strongly colocalized with nsp8 and nsp12, both well-defined members of the RTC, but not the membrane (M) protein, involved in virus assembly. Finally, we found that nsp15 interacted with RTC-associated proteins nsp8 and nsp12 during infection, and this interaction was RNA independent. From this, we conclude that nsp15 localizes and interacts with CoV proteins in the RTC, suggesting it plays a direct or indirect role in virus replication. Furthermore, the use of
in situ
epitope tags could be used to determine novel nsp-nsp interactions in coronaviruses.

IMPORTANCE
Despite structural and biochemical data demonstrating that the coronavirus nsp15 protein contains an endoribonuclease domain, its precise function during coronavirus infection remains unknown. In this work, we created a novel
in situ
tagged form of nsp15 to study interactions and localization during infection. This
in situ
tag was tolerated by MHV and did not affect viral replication. Utilizing this tag, we established that nsp15 localized to sites of replication but not sites of assembly throughout infection. Furthermore, we found that nsp15 interacted with the putative viral primase nsp8 and polymerase nsp12 during CoV infection. The strong association of nsp15 with replication complexes and interactions with replicative CoV enzymes suggest nsp15 is involved in CoV replication. These data and tools developed in this study help elucidate the function of nsp15 during infection and may be used to uncover other novel viral protein interactions.
Characteristics of the local cutaneous sensory thermoneutral zone
Authors: Filingeri Davide , Zhang Hui , Arens Edward A. 
Published: February 2017

Skin temperature detection thresholds have been used to measure human cold and warm sensitivity across the temperature continuum. They exhibit a sensory zone within which neither warm nor cold sensations prevail. This zone has been widely assumed to coincide with steady-state local skin temperatures between 32 and 34°C, but its underlying neurophysiology has been rarely investigated. In this study we employ two approaches to characterize the properties of sensory thermoneutrality, testing for each whether neutrality shifts along the temperature continuum depending on adaptation to a preceding thermal state. The focus is on local spots of skin on the palm. Ten participants (age: 30.3 ± 4.8 yr) underwent two experiments. Experiment 1 established the cold-to-warm inter-detection threshold range for the palm’s glabrous skin and its shift as a function of 3 starting skin temperatures (26, 31, or 36°C). For the same conditions, experiment 2 determined a thermally neutral zone centered around a thermally neutral point in which thermoreceptors’ activity is balanced. The zone was found to be narrow (~0.98 to ~1.33°C), moving with the starting skin temperature over the temperature span 27.5–34.9°C (Pearson r = 0.94; P < 0.001). It falls within the cold-to-warm inter-threshold range (~2.25 to ~2.47°C) but is only half as wide. These findings provide the first quantitative analysis of the local sensory thermoneutral zone in humans, indicating that it does not occur only within a specific range of steady-state skin temperatures (i.e., it shifts across the temperature continuum) and that it differs from the inter-detection threshold range both quantitatively and qualitatively. These findings provide insight into thermoreception neurophysiology. NEW & NOTEWORTHY Contrary to a widespread concept in human thermoreception, we show that local sensory thermoneutrality is achievable outside the 32–34°C skin temperature range. We propose that sensory adaption underlies a new mechanism of temperature integration. Also, we have developed from vision research a new quantitative test addressing the balance in activity of cutaneous cold and warm thermoreceptors. This could have important clinical (assessment of somatosensory abnormalities in neurological disease) and applied (design of personal comfort systems) implications.
Emerging Role of MicroRNAs as Liquid Biopsy Biomarkers in Gastrointestinal Cancers
Authors: Shigeyasu Kunitoshi , Toden Shusuke , Zumwalt Timothy J. , Okugawa Yoshinaga , Goel Ajay 
Published: February 2017

Abstract
Cancer has emerged as a leading cause of mortality worldwide, claiming more than 8 million lives annually. Gastrointestinal cancers account for about 35% of these mortalities. Recent advances in diagnostic and treatment strategies have reduced mortality among patients with gastrointestinal cancer, yet a significant number of patients still develop late-stage cancer, where treatment options are inadequate. Emerging interests in “liquid biopsies” have encouraged investigators to identify and develop clinically relevant noninvasive genomic and epigenomic signatures that can be exploited as biomarkers capable of detecting premalignant and early-stage cancers. In this context, microRNAs (miRNA), which are small, noncoding RNAs that are frequently dysregulated in cancers, have emerged as promising entities for such diagnostic purposes. Even though the future looks promising, current approaches for detecting miRNAs in blood and other biofluids remain inadequate. This review summarizes existing efforts to exploit circulating miRNAs as cancer biomarkers and evaluates their potential and challenges as liquid biopsy–based biomarkers for gastrointestinal cancers. Clin Cancer Res; 23(10); 2391–9. ©2017 AACR.
Musashi RNA-Binding Proteins as Cancer Drivers and Novel Therapeutic Targets
Authors: Kudinov Alexander E. , Karanicolas John , Golemis Erica A. , Boumber Yanis 
Published: February 2017

Abstract
Aberrant gene expression that drives human cancer can arise from epigenetic dysregulation. Although much attention has focused on altered activity of transcription factors and chromatin-modulating proteins, proteins that act posttranscriptionally can potently affect expression of oncogenic signaling proteins. The RNA-binding proteins (RBP) Musashi-1 (MSI1) and Musashi-2 (MSI2) are emerging as regulators of multiple critical biological processes relevant to cancer initiation, progression, and drug resistance. Following identification of Musashi as a regulator of progenitor cell identity in Drosophila, the human Musashi proteins were initially linked to control of maintenance of hematopoietic stem cells, then stem cell compartments for additional cell types. More recently, the Musashi proteins were found to be overexpressed and prognostic of outcome in numerous cancer types, including colorectal, lung, and pancreatic cancers; glioblastoma; and several leukemias. MSI1 and MSI2 bind and regulate the mRNA stability and translation of proteins operating in essential oncogenic signaling pathways, including NUMB/Notch, PTEN/mTOR, TGFβ/SMAD3, MYC, cMET, and others. On the basis of these activities, MSI proteins maintain cancer stem cell populations and regulate cancer invasion, metastasis, and development of more aggressive cancer phenotypes, including drug resistance. Although RBPs are viewed as difficult therapeutic targets, initial efforts to develop MSI-specific inhibitors are promising, and RNA interference–based approaches to inhibiting these proteins have had promising outcomes in preclinical studies. In the interim, understanding the function of these translational regulators may yield insight into the relationship between mRNA expression and protein expression in tumors, guiding tumor-profiling analysis. This review provides a current overview of Musashi as a cancer driver and novel therapeutic target. Clin Cancer Res; 23(9); 2143–53. ©2017 AACR.
The Tsar’s Two Bodies
Authors: Keenan Edward L. , Martin Russell E. 
Published: February 2017

Originally delivered as a lecture at Columbia University in the fall of 1974, this classic text has circulated among Professor Edward L. Keenan’s students and colleagues for decades, but has never been published. The lecture draws upon the author’s textual work, including his then recently published book challenging the provenance of the Correspondence attributed to Tsar Ivan IV and Prince Andrei Kurbskii, to explore how his conclusions about this and other texts impinge on the received historiographical traditions about power, dynasty, and the life and reign of Ivan IV “the Terrible.” The lecture employs the model developed by Ernst Kantorowicz in his The King’s Two Bodies to understand the biographical details of Ivan IV, including the results of the autopsy of Ivan’s remains. The lecture provides an early, yet penetrating glimpse into the thinking of one of the most influential historians of Russia in the sixteenth century and of the reign of Ivan the Terrible.
Evaluation of a proper cutoff value on quantitative dual-energy perfusion CT for the assessment of acute pulmonary thromboembolism
Authors: Kunihiro Yoshie , Okada Munemasa , Matsunaga Naofumi 
Published: February 2017

Background The cutoff value for assessing the severity of acute pulmonary thromboembolism (PTE) using relative volumetric evaluations of dual-energy perfusion computed tomography (DEpCT) is unclear. Purpose To determine the proper cutoff value for determining the severity of PTE using DEpCT volumetry. Material and Methods A total of 185 patients with venous thromboembolism were included in this study, of whom 61 were diagnosed with acute PTE. DEpCT images were three-dimensionally reconstructed at the following attenuation ranges: 1–2 HU (V2), 1–10 HU (V10), and 1–120 HU (V120). The ratios of low perfusion areas associated with each threshold range per V120 were also calculated, and the relative ratios were expressed as %V2 to %V10. These values were compared with factors indicating the severity of PTE, including the pulmonary arterial pressure, heart rate, CT angiographic obstruction index (CTOI), and right/left ventricular diameter ratio (RV/LV). Results The area under the curve (AUC) of %V2 was highest (0.783) among these values (95% confidence interval, 0.710–0.856) based on the presence of IPCs. The %V2 showed moderate correlations with CTOI (r = 0.36, P = 0.005) and RV/LV (r = 0.36, P = 0.004) in the patients with acute PTE. Conclusion Volumetric evaluations of DEpCT images using the lowest attenuation threshold range (1–2 HU) exhibit the best correlation with factors suggesting the severity of acute PTE.
Biomechanical Comparison of Anterolateral Procedures Combined With Anterior Cruciate Ligament Reconstruction
Authors: Inderhaug Eivind , Stephen Joanna M. , Williams Andy , Amis Andrew A. 
Published: February 2017

Background: Anterolateral soft tissue structures of the knee have a role in controlling anterolateral rotational laxity, and they may be damaged at the time of anterior cruciate ligament (ACL) ruptures. Purpose: To compare the kinematic effects of anterolateral operative procedures in combination with intra-articular ACL reconstruction for combined ACL plus anterolateral–injured knees. Study Design: Controlled laboratory study. Methods: Twelve cadaveric knees were tested in a 6 degrees of freedom rig using an optical tracking system to record the kinematics through 0° to 90° of knee flexion with no load, anterior drawer, internal rotation, and combined loading. Testing was first performed in ACL-intact, ACL-deficient, and combined ACL plus anterolateral–injured (distal deep insertions of the iliotibial band and the anterolateral ligament [ALL] and capsule cut) states. Thereafter, ACL reconstruction was performed alone and in combination with the following: modified MacIntosh tenodesis, modified Lemaire tenodesis passed both superficial and deep to the lateral collateral ligament, and ALL reconstruction. Anterolateral grafts were fixed at 30° of knee flexion with both 20 and 40 N of tension. Statistical analysis used repeated-measures analyses of variance and paired t tests with Bonferroni adjustments. Results: ACL reconstruction alone failed to restore native knee kinematics in combined ACL plus anterolateral–injured knees ( P < .05 for all). All combined reconstructions with 20 N of tension, except for ALL reconstruction ( P = .002-.01), restored anterior translation. With 40 N of tension, the superficial Lemaire and MacIntosh procedures overconstrained the anterior laxity in deep flexion. Only the deep Lemaire and MacIntosh procedures—with 20 N of tension—restored rotational kinematics to the intact state ( P > .05 for all), while the ALL underconstrained and the superficial Lemaire overconstrained internal rotation. The same procedures with 40 N of tension led to similar findings. Conclusion: In a combined ACL plus anterolateral–injured knee, ACL reconstruction alone failed to restore intact knee kinematics. The addition of either the deep Lemaire or MacIntosh tenodesis tensioned with 20 N, however, restored native knee kinematics. Clinical Relevance: The current study indicates that unaddressed anterolateral injuries, in the presence of an ACL deficiency, result in abnormal knee kinematics that is not restored if only treated with intra-articular ACL reconstruction. Both the modified MacIntosh and modified deep Lemaire tenodeses (with 20 N of tension) restored native knee kinematics at time zero.
Inflammatory Microenvironment Persists After Bone Healing in Intra-articular Ankle Fractures
Authors: Adams Samuel B. , Leimer Elizabeth M. , Setton Lori A. , Bell Richard D. , Easley Mark E. , Huebner Janet L. , Stabler Thomas V. , Kraus Virginia B. , Olson Steven A. , Nettles Dana L. 
Published: February 2017

Background: Post-traumatic osteoarthritis (PTOA) is responsible for the majority of cases of ankle arthritis. While acute and end-stage intra-articular inflammation has previously been described, the state of the joint between fracture healing and end-stage PTOA remains undefined. This study characterized synovial fluid (SF) composition of ankles after bone healing of an intra-articular fracture to identify factors that may contribute to the development of PTOA. Methods: Of an original 21 patients whose SF was characterized acutely following intra-articular ankle fractures, 7 returned for planned hardware (syndesmotic screw) removal after bone healing (approximately 6 months) and consented to a second bilateral SF collection. SF concentrations of 15 cytokines and matrix metalloproteinases (MMPs) and 2 markers each of cartilage catabolism (CTXII and glycosaminoglycan) and hemarthrosis (biliverdin and bilirubin) were compared for previously fractured and contralateral, uninjured ankles from the same patient. Analysis was also performed to determine the effect of the number of fracture lines and involvement of soft tissue on SF composition. Results: Interleukin (IL)-6, IL-8, MMP-1, MMP-2, and MMP-3 were significantly elevated in the SF from healed ankles compared to matched contralateral uninjured ankles at approximately 6 months after fracture. There were no differences in markers of cartilage catabolism or hemarthrosis. Only IL-1α was affected by the number of fracture lines while differences were not detected for other analytes or with respect to the involvment of soft tissue. Conclusions: Sustained intra-articular inflammation, even after complete bone healing, was suggested by elevations of pro-inflammatory cytokines (IL-6 and IL-8). In addition, elevated concentrations of MMPs were also noted and were consistent with a persistent inflammatory environment. This study suggests new evidence of persistent intra-articular inflammation after intra-articular ankle fracture healing and suggests potential mediators for PTOA development. Clinical Relevance: This work may be relevant to the clinical diagnosis and treatment of post-traumatic osteoarthritis.
Health-related quality of life outcomes from botulinum toxin treatment in hemifacial spasm
Authors: Weiss Daniel , Sturm Justine , Hieber Leonhard , Börtlein Axel , Mayr Ingo , Appy Matthias , Kühnler Benedicta , Buchthal Joachim , Dippon Christian , Arnold Guy , Wächter Tobias 
Published: February 2017

Background: Although botulinum neurotoxin (BoNT) injections may alleviate involuntary muscle contractions in hemifacial spasm substantially, it is less clear whether the motor effect would translate into improvements of health-related quality of life (HR-QoL). Methods: In this open-label clinical observational study, we characterized outcomes on HR-QoL in terms of the EuroQol (EQ-5D-5L) from BoNT in a prospective cohort of patients with hemifacial spasm ( n = 73). Additionally, we characterized appendicular motor and nonmotor signs on motor symptom improvement, depressive symptoms, pain and sleep quality. Patients were assessed at the end of a regular 3-month period from last injection (timepoint1) and 4 weeks after the reinjection of BoNT (timepoint2). Results: Patients showed improved HR-QoL on the EQ-VAS (visual analogue scale) at timepoint2 compared with timepoint1. Moreover, we identified, that impairments in HR-QoL at timepoint1 correlated with life satisfaction and depressive symptoms, respectively. However, these associated variables did not predict the therapeutic effect. Instead, EQ-VAS at timepoint1 accounted for 34.5% of the variance of EQ-VAS improvement expressed as the difference between timepoint2 and timepoint1. Conclusion: Our study supports HR-QoL improvements in hemifacial spasm and the value of generic HR-QoL measures to estimate therapeutic outcome. However, the findings should be considered descriptive, and future high quality trials are needed for confirmatory purposes in order to refine treatment referral in hemifacial spasm with respect to QoL.
Comparison of Guy’s vs S.T.O.N.E. nephrolithometry scoring systems in predicting the success rate of PCNL
Authors: Rathee VS , HC Vivek , Khan Sartaj Wali , Singh AK , Shukla Pushpender Kumar , Verma Ashish , Trivedi Sameer , Dwivedi Udai Shankar 
Published: February 2017

Introduction and objective: The aim of the present study was to compare the accuracy of the Guy’s and S.T.O.N.E. scoring systems in predicting percutaneous nephrolithotomy (PCNL) outcomes. Methods: This prospective study was carried out in the Department of Urology, Institute of Medical Sciences, Banaras Hindu University, Varanasi, between August 2013 and September 2015 in collaboration with the Department of Radiodiagnosis and Imaging. A total of 100 cases of renal calculi operated on with PCNL were considered in the study on the basis of CT morphodensitometry. The study was approved by the institute’s ethical committee. Guy’s and S.T.O.N.E. scoring systems were calculated. Regression analysis and ROC curves were performed. Results: A total of 100 PCNLs were operated on. The overall stone-free rate was 90% with a complication rate of 17%. When compared to patients with residual fragments, stone-free patients had significantly lower Guy’s grade (3.5 vs 2; p < 0.001) and S.T.O.N.E. score (9.1 vs 7.5; p = 0.023). Logistic regression analysis showed that both Guy’s and S.T.O.N.E. systems were significantly associated with stone-free status, OR 0.96 ( p < 0.001) and OR 0.94 ( p < 0.001), respectively. Furthermore, both scoring systems were significantly associated with estimated blood loss ( p = 0.02 and p = 0.004). There was good correlation between both scoring systems and operative time ( r = 0.2, p < 0.002 and r = 0.3, p < 0.002) and length of hospital stay ( r = 0.1, p = 0.002 and r = 0.2, p < 0.002). There were significant associations between both scoring systems and complications ( p < 0.001 and p = 0.004). There was no significant difference in the areas under the curves for the Guy’s and S.T.O.N.E. scoring systems (0.96 (95% CI 0.92–0.99) vs 0.94 (95% CI 0.89–1.001); p < 0.001). Conclusion: Both Guy’s and S.T.O.N.E scoring systems have comparable accuracies in predicting post-PCNL stone-free status. Other factors not included in either scoring system may need to be incorporated in the future to increase their accuracy.
Finger and foot tapping as alternative outcomes of upper and lower extremity function in multiple sclerosis
Authors: Tanigawa Makoto , Stein Jason , Park John , Kosa Peter , Cortese Irene , Bielekova Bibiana 
Published: February 2017

Background While magnetic resonance imaging contrast-enhancing lesions represent an excellent screening tool for disease-modifying treatments in relapsing–remitting multiple sclerosis (RRMS), this biomarker is insensitive for testing therapies against compartmentalized inflammation in progressive multiple sclerosis (MS). Therefore, alternative sensitive outcomes are needed. Using machine learning, clinician-acquired disability scales can be combined with timed measures of neurological functions such as walking speed (e.g. 25-foot walk; 25FW) or fine finger movements (e.g. 9-hole peg test; 9HPT) into sensitive composite clinical scales, such as the recently developed combinatorial, weight-adjusted disability scale (CombiWISE). Ideally, these complementary simplified measurements of certain neurological functions could be performed regularly at patients’ homes using smartphones. Objectives We asked whether tests amenable to adaptation to smartphone technology, such as finger and foot tapping have comparable sensitivity and specificity to current non-clinician-acquired disability measures. Results We observed that finger and foot tapping can differentiate RRMS and progressive MS in a cross-sectional study and can also measure yearly and two-year disease progression in the latter, with better power (based on z-scores) in comparison to currently utilized 9HPT and 25FW. Conclusions Replacing the 9HPT and 25FW with simplified tests broadly adaptable to smartphone technology may enhance the power of composite scales for progressive MS.