bluuebunny/crossref_metadata_2025 huggingface dataset:
Influence of Androgen Receptor CAG Polymorphism on Sexual Function Recovery after Testosterone Therapy in Late-Onset Hypogonadism
Authors: Tirabassi Giacomo , Corona Giovanni , Biagioli Andrea , Buldreghini Eddi , delli Muti Nicola , Maggi Mario , Balercia Giancarlo
Published: December 2014

Introduction

Androgen receptor (AR) CAG polymorphism has been found to influence sexual function. However, no study has evaluated its potential to condition sexual function recovery after testosterone replacement therapy (TRT) in a large cohort of hypogonadic subjects.

Aim

To evaluate the role of this polymorphism in sexual function improvement after TRT in late-onset hypogonadism (LOH).

Methods

Seventy-three men affected by LOH were retrospectively considered. Evaluations were performed before TRT started (time 0) and before the sixth undecanoate testosterone injection.

Main Outcome Measures

International Index of Erectile Function (IIEF) questionnaire (erectile function [EF], orgasmic function [OF], sexual desire [SD], intercourse satisfaction [IS], overall satisfaction [OS], and total IIEF-15 score); total and free testosterone and estradiol; AR gene CAG repeat number.

Results

TRT induced a significant increase in total and free testosterone and estradiol. All IIEF domains significantly improved after TRT. AR CAG repeats negatively and significantly correlated with all the variations (Δ-) of sexual function domains, except for Δ-OS. Conversely, Δ-total testosterone was found to be positively and significantly correlated with sexual function domain variations, except for Δ-IS and Δ-OS. Δ-estradiol did not correlate significantly with any of the variations of sexual function domains. After inclusion in generalized linear models, the number of AR gene CAG triplets was found to be independently and negatively associated with Δ-EF, Δ-SD, Δ-IS, and Δ-Total IIEF-15 score, whereas Δ-total testosterone was independently and positively associated with Δ-EF, Δ-OF, Δ-SD, and Δ-Total IIEF-15 score. However, after including time 0 total testosterone in the model, AR gene CAG triplets remained independently and negatively associated only with Δ-EF and Δ-Total IIEF-15 score, whereas Δ-total testosterone was independently and positively associated only with Δ-EF.

Conclusions

Longer length of AR gene CAG repeat tract seems to lower TRT-induced improvement of sexual function in LOH.
A neuronal DNA damage response is detected at the earliest stages of Alzheimer's neuropathology and correlates with cognitive impairment in the Medical Research Council's Cognitive Function and Ageing Study ageing brain cohort
Authors: Simpson Julie E. , Ince Paul G. , Matthews Fiona E. , Shaw Pamela J. , Heath Paul R. , Brayne Carol , Garwood Claire , Higginbottom Adrian , Wharton Stephen B. ,   MRC Cognitive Function and Ageing Neuropathology Study Group
Published: December 2014

Aims

Population‐based studies have shown that approximately 20% of the ageing population (aged 65 years and over) with dementia have little or no classical Alzheimer‐type neuropathology. Cumulative DNA damage and a reduced capacity of DNA repair may result in neuronal dysfunction and contribute to cognitive impairment independent of Alzheimer‐type pathology in the ageing brain.

Methods

We investigated expression of the DNA damage response (DDR)‐associated molecules γH2AX and DNA‐PKcs using immunohistochemistry and western blotting, and senescence‐associated β‐galactosidase in the frontal association neocortex of cases with low levels of Alzheimer‐type pathology (Braak & Braak stage 0–II), and explored their relationship to cognitive impairment in a population‐representative sample from the Medical Research Council's Cognitive Function and Ageing Study cohort.

Results

Increases in both γH2AX+ (rs = −0.36, P = 0.025) and DNA‐PKcs+ (rs = −0.39, P = 0.01) neuronal counts were associated with a lower Mini‐Mental State Examination score. Increasing levels of senescence associated‐β‐gal+ pyramidal neurones were weakly associated with the total number of DNA‐PKcs+ neurones (P = 0.08), but not with traditional senescence‐associated signalling molecules, including p53 and p16.

Conclusion

The association between the neuronal DDR and cognitive impairment, independent of AD pathology in the ageing brain, may be suggestive of a causal link via neuronal dysfunction.
Early‐life stress selectively affects gastrointestinal but not behavioral responses in a genetic model of brain–gut axis dysfunction
Authors: Hyland N. P. , O'Mahony S. M. , O'Malley D. , O'Mahony C. M. , Dinan T. G. , Cryan J. F.
Published: December 2014

Background

Early‐life stress and a genetic predisposition to display an anxiety‐ and depressive‐like phenotype are associated with behavioral and gastrointestinal (GI) dysfunction. Animals exposed to early‐life stress, and those genetically predisposed to display anxiety or depressive behaviors, have proven useful tools in which to study stress‐related GI disorders, such as irritable bowel syndrome (IBS). IBS is a heterogeneous disorder, and likely a consequence of both genetic and environmental factors. However, the combined effects of early‐life stress and a genetic predisposition to display anxiety‐ and depression‐like behaviors on GI function have not been investigated.

Methods

We assessed the effect of maternal separation (MS) on behavioral and GI responses in WKY animals relative to a normo‐anxious reference strain.

Key Results

Both non‐separated (NS) WKY and WKY‐MS animals displayed anxiety‐like responses in the open‐field test and depressive‐like behaviors in the forced swim test relative to Sprague–Dawley rats. However, MS had no further influence on anxiety‐ and depressive‐like behaviors exhibited by this stress‐prone rat strain. Similarly, corticosterone levels measured after the OFT were insensitive to MS in WKY animals. However, WKY‐MS displayed significantly increased colonic visceral hypersensitivity, fecal output, and altered colonic cholinergic sensitivity.

Conclusions & Inferences

Our data suggest that early‐life stress, on the background of a genetic predisposition to display an anxiety‐ and depressive‐like phenotype, selectively influences GI function rather than stress‐related behaviors. Thus, our findings highlight the importance of genetic predisposition on the outcome of early‐life adversity on GI function.
Atypical hemolytic uremic syndrome: Korean pediatric series
Authors: Lee Jiwon M. , Park Young Seo , Lee Joo Hoon , Park Se Jin , Shin Jae Il , Park Yong‐Hoon , Yoo Kee Hwan , Cho Min Hyun , Kim Su‐Young , Kim Seong Heon , Namgoong Mee Kyung , Lee Seung Joo , Lee Jun Ho , Cho Hee Yeon , Han Kyoung Hee , Kang Hee Gyung , Ha Il Soo , Bae Jun‐Seok , Kim Nayoung K. D. , Park Woong‐Yang , Cheong Hae Il
Published: December 2014

Background

Atypical hemolytic uremic syndrome (aHUS) is a rare disease with a genetic predisposition. Few studies have evaluated the disease in the Asian population. We studied a Korean pediatric cohort to delineate the clinical characteristics and genotypes.

Methods

A multicenter cohort of 51 Korean children with aHUS was screened for mutations using targeted exome sequencing covering 46 complement related genes. Anti‐complement‐factor‐H autoantibody (anti‐CFH) titers were measured. Multiplex ligation‐dependent probe amplification assay was performed to detect deletions in the complement factor‐H related protein genes (CFHR) in the patients as well as in 100 healthy Korean controls. We grouped the patients according to etiology and compared the clinical features using Mann–Whitney U‐test and chi‐squared test.

Results

Fifteen patients (group A, 29.7%) had anti‐CFH, and mutations were detected in 11 (group B, 21.6%), including one with combined mutations. The remaining 25 (group C, 49.0%) were negative for both. The prevalence of anti‐CFH was higher than the worldwide level. Group A had a higher onset age than group B, although the difference was not significant. Group B had the worst renal outcome. Gene frequencies of homozygous CFHR1 deletion were 73.3%, 2.7% and 1% in group A, group B + C and the control, respectively.

Conclusions

The incidence of anti‐CFH in the present Korean aHUS cohort was high. Clinical outcomes largely conformed to the previous reports. Although the sample size was limited, this cohort provides a reassessment of clinicogenetic features of aHUS in Korean children.
Seated movement indexes emotion and its regulation in posttraumatic stress disorder
Authors: Woodward Steven H. , Shurick Ashley A. , Alvarez Jennifer , Kuo Janice , Nonyieva Yuliana , Blechert Jens , McRae Kateri , Gross James J.
Published: December 2014

This paper reports the results of a study that administered an emotion regulation task to Operation Enduring Freedom/Operation Iraqi Freedom veterans diagnosed with posttraumatic stress disorder (27) and to healthy controls (23). Seated movement and postural responses were transduced with a sensitive accelerometer attached to the underside of a low‐mass cantilevered chair. Consistent with prior studies in which subjects stood on force plates, aversive photographs induced attenuation of nondirectional movement in patients and controls. Regarding seated postural responses, controls leaned towards neutral photographs and away from aversive ones, while participants with PTSD did the opposite. Regulation had no impact on seated movement but was associated with a seated postural withdrawal from the computer screen.
Evaluation of the Coaches Educators training implementation of the PAPA project: A comparison between Norway and France
Authors: Van Hoye A. , Larsen T. , Sovik M. , Wold B. , Heuzé J‐P. , Samdal O. , Ommundsen Y. , Sarrazin P.
Published: December 2014

This study cross‐culturally compares the implementation process of the Empowering Coaching™ training program to grassroots coaches (GCs) by trained Coach Educators (CEs) from the Promoting Adolescent Physical Activity project in Norway and France. This program targets children's health through coach training to create a more positive environment. Using the RE‐AIM framework, indicators of implementation processes at staff level were defined for adoption, implementation, and maintenance. CEs (n = 18) were interviewed about their professional use of the training, and video‐filmed delivering the intervention trial workshops. GCs (n = 185) responded to a questionnaire after the workshops. Results showed that CEs were keen to participate in this project (16/18; 89%), to stay involved (12/16; 75%), and to diffuse the principles after the research project (12/12; 100%). CE training seemed effective as results indicated that CEs applied the principles of the program during the workshops and covered a high degree of the content (78%). Cross‐cultural differences were found as the French Football Federation CE dropped out of the program, as Norwegian CEs delivered the content with higher fidelity and had higher enjoyment and pace scores than French CEs. More indicators are needed to use the RE‐AIM framework for cross‐cultural comparison.
Objectively measured physical activity in Danish after‐school cares: Does sport certification matter?
Authors: Domazet S. L. , Møller N. C. , Støckel J. T. , Ried‐Larsen M.
Published: December 2014

Inactivity and more sedentary time predominate the daily activity level of many of today's children. In Denmark, certified sport after‐school cares have been established in order to increase children's daily physical activity (PA) level. This cross‐sectional study aimed to investigate the activity level among participants in certified sport after‐school cares vs regular after‐school cares. The study was carried out in 2011 in 10 after‐school cares (5 sport/5 regular) throughout Denmark, whereof 475 children aged 5–11 years participated. PA level was assessed using Actigraph GT3X and GT3X+ activity monitors worn by the children for at least 8 consecutive days. Anthropometry and cardiorespiratory fitness were measured as well. A multivariate regression analysis was carried out to check for the differences in the PA level across the two care systems. However, there did not appear to be any differences in overall PA or in time‐specific day parts (e.g., during after‐school care). The activity levels were quite similar across after‐school cares and were mutually high during time spent in the care facility.
Young athletes' awareness and monitoring of anti‐doping in daily life: Does motivation matter?
Authors: Chan D. K. C. , Donovan R. J. , Lentillon‐Kaestner V. , Hardcastle S. J. , Dimmock J. A. , Keatley D. A. , Hagger M. S.
Published: December 2014

This study was a preliminarily investigation into the prevention of unintentional doping on the basis of self‐determination theory (SDT). Specifically, we examined the relationship between athletes' motives for doping avoidance and their behavior when offered an unfamiliar food product. Participants were young Australian athletes (n = 410) that were offered a free lollipop prior to completing a questionnaire. It was noted whether participants refused to take or eat the lollipop and whether they read the ingredients of the lollipop. The questionnaire assessed autonomous and controlled forms of motivation, amotivation, doping intentions, and adherence regarding doping avoidance behaviors. The results showed that young athletes who adopted controlled reasons to avoid doping in sport (e.g., not getting caught) tended to report higher adherence to behaviors related to avoiding and monitoring banned substances, whereas those who adopted autonomous reasons (e.g., anti‐doping being consistent with life goals) appeared to be more willing to read the ingredients of the provided food. The significant interaction effect between autonomous and controlled motivation indicated that autonomous motivation was more predictive to doping intention for athletes with low controlled motivation. It is concluded that SDT may help understand the motivational processes of the prevention of unintentional doping in sport.
Early Parental Positive Behavior Support and Childhood Adjustment: Addressing Enduring Questions with New Methods
Authors: Waller Rebecca , Gardner Frances , Dishion Thomas , Sitnick Stephanie L. , Shaw Daniel S. , Winter Charlotte E. , Wilson Melvin
Published: December 2014

A large literature provides strong empirical support for the influence of parenting on child outcomes. The current study addresses enduring research questions testing the importance of early parenting behavior to children's adjustment. Specifically, we developed and tested a novel multi‐method observational measure of parental positive behavior support at age 2. Next, we tested whether early parental positive behavior support was related to child adjustment at school age, within a multi‐agent and multi‐method measurement approach and design. Observational and parent‐reported data from mother–child dyads (N = 731; 49 percent female) were collected from a high‐risk sample at age 2. Follow‐up data were collected via teacher report and child assessment at age 7.5. The results supported combining three different observational methods to assess positive behavior support at age 2 within a latent factor. Further, parents' observed positive behavior support at age 2 predicted multiple types of teacher‐reported and child‐assessed problem behavior and competencies at 7.5 years old. Results supported the validity and predictive capability of a multi‐method observational measure of parenting and the importance of a continued focus on the early years within preventive interventions.
Urea retranslocation from senescing Arabidopsis leaves is promoted by DUR3‐mediated urea retrieval from leaf apoplast
Authors: Bohner Anne , Kojima Soichi , Hajirezaei Mohammad , Melzer Michael , von Wirén Nicolaus
Published: December 2014

In plants, urea derives either from root uptake or protein degradation. Although large quantities of urea are released during senescence, urea is mainly seen as a short‐lived nitrogen (N) catabolite serving urease‐mediated hydrolysis to ammonium. Here, we investigated the roles of DUR3 and of urea in N remobilization. During natural leaf senescence urea concentrations and DUR3 transcript levels showed a parallel increase with senescence markers like ORE1 in a plant age‐ and leaf age‐dependent manner. Deletion of DUR3 decreased urea accumulation in leaves, whereas the fraction of urea lost to the leaf apoplast was enhanced. Under natural and N deficiency‐induced senescence DUR3 promoter activity was highest in the vasculature, but was also found in surrounding bundle sheath and mesophyll cells. An analysis of petiole exudates from wild‐type leaves revealed that N from urea accounted for >13% of amino acid N. Urea export from senescent leaves further increased in ureG‐2 deletion mutants lacking urease activity. In the dur3 ureG double insertion line the absence of DUR3 reduced urea export from leaf petioles. These results indicate that urea can serve as an early metabolic marker for leaf senescence, and that DUR3‐mediated urea retrieval contributes to the retranslocation of N from urea during leaf senescence.
Effect of tiletamine–zolazepam on the intraocular pressure of the dog
Authors: Jang Min , Park Sangwan , Son Won‐gyun , Jo Sang‐min , Hwang Hyeshin , Seo Kangmoon , Lee Inhyung
Published: November 2014

Objective

To determine the effect of tiletamine–zolazepam (TZ) on canine intraocular pressure (IOP).

Animals studied

Six healthy dogs without ocular abnormalities were used in this study.

Procedures

The study was carried out as a crossover experimental trial with a 7‐day interval between treatments. TZ combination was administered intravenously (IV) at a dose of 5, 10, and 20 mg/kg (TZ5, TZ10, and TZ20, respectively). Following preanesthetic baseline readings of IOP, each dog received IV TZ and then the IOP values were measured every 10 min for 40 min in all treatments.

Results

The baseline IOP values (mean ± standard deviation) for TZ5, TZ10, and TZ20 were 12.7 ± 0.8, 14.4 ± 1.2, and 15.3 ± 1.7 mmHg, and each IOP changed to 11.1 ± 1.1, 13.1 ± 1.4 and 13.5 ± 1.7 mmHg after intravenous administration of each TZ treatment, respectively. However, there were no statistical differences between baseline and post‐treatment values.

Conclusions

The TZ combination had no clinically significant effect on IOP of the dog. This could be an option for induction or surgical procedures in dogs with ophthalmic problems when an increase in IOP is undesirable.
Dynamic Gains Differentiator for Hydraulic System Control
Authors: Sidhom L. , Brun X. , Smaoui M. , Bideaux E. , Thomasset D.
Published: December 2014

This paper deals with online numerical differentiation of a noisy time signal where new higher order sliding mode differentiators are proposed. The key point of these algorithms is to include a dynamic on the differentiator parameters. These dynamics tune-up automatically the algorithm gains in real-time. Convergence properties of the new schemes are derived using a Lyapunov approach. Their effectiveness is illustrated via simulations and experimental tests, where comparative studies are performed between classical schemes and the new ones. Such algorithms are also used in the feedback control of an electrohydraulic system.
Enhanced Computational Fluid Dynamics Modeling and Laser Doppler Anemometer Measurements for the Air-Flow in an Aero-engine Front Bearing Chamber—Part I
Authors: Aidarinis J. , Goulas A.
Published: December 2014

Modern aero-engine development requires also a gradual increase in the overall effectiveness of lubrication systems. This particularly applies to bearing chambers where a complex two-phase flow is formed by the interaction of the sealing air and the lubrication oil. It is important to increase the level of understanding of the flow field inside the bearing chamber and to develop engineering tools in order to optimize its design and improve its performance. To achieve this, an experimental and a computational study of the whole front bearing chamber were carried out for a range of shaft rotational speeds and sealing air mass flow. The experimental measurements of the air velocity inside the chamber were carried out using a laser Doppler anemometer (LDA) in two-phase air/oil-flow conditions. The experimental facility is a 1:1 scale model of the front bearing chamber of an aero-engine. Computational 3D modeling of the bearing chamber was performed. The bearing gap and the presence of lubrication oil were modeled as an anisotropic porous medium with functions relating the pressure loss of the air coming through the gap and the tangential component of velocity of the air exiting the gap of the ball bearing with the air-flow rate through the gap and the rotational speed of the shaft. The methodology to obtain the above mentioned functions and the results of the detailed study are given (Aidarinis, J., and Goulas, A., 2014, “Enhanced CFD Modeling and LDA Measurements for the Air-Flow in an Aero Engine Front Bearing Chamber: Part II,” ASME Paper No. GT2014-26062). The enhanced computational model of the chamber implementing the law of pressure drop of the “lubricated” bearing and the function of modeling the tangential velocity of the air exiting the bearing was used to calculate the flow field for the full range of the measurements. A limiting curve dividing the operational map of the bearing chamber into two areas was predicted. Large vortical and swirling structures dominate the flow and they vary in size according to the position of the operation point relative to the limiting curve. Operation above the limiting curve leads to flow classified as type I with air going through the ball bearing while for operation below the limiting curve line the flow is classified as type II, there is no air-flow through the bearing gap.
Spatial atomic layer deposition on flexible substrates using a modular rotating cylinder reactor
Authors: Sharma Kashish , Hall Robert A. , George Steven M.
Published: December 2014

Spatial atomic layer deposition (ALD) is a new version of ALD based on the separation of reactant gases in space instead of time. In this paper, the authors present results for spatial ALD on flexible substrates using a modular rotating cylinder reactor. The design for this reactor is based on two concentric cylinders. The outer cylinder remains fixed and contains a series of slits. These slits can accept a wide range of modules that attach from the outside. The modules can easily move between the various slit positions and perform precursor dosing, purging, or pumping. The inner cylinder rotates with the flexible substrate and passes underneath the various spatially separated slits in the outer cylinder. Trimethyl aluminum and ozone were used to grow Al2O3 ALD films at 40 °C on metallized polyethylene terephthalate (PET) substrates to characterize this spatial ALD reactor. Spectroscopic ellipsometry measurements revealed a constant Al2O3 ALD growth rate of 1.03 Å/cycle with rotation speeds from 40 to 100 RPM with the outer cylinder configured for one Al2O3 ALD cycle per rotation. The Al2O3 ALD growth rate then decreased at higher rotation rates for reactant residence times < 5 ms. The Al2O3 ALD films were also uniform to within <1% across the central portion of metallized PET substrate. Fixed deposition time experiments revealed that Al2O3 ALD films could be deposited at 2.08 Å/s at higher rotation speeds of 175 RPM. Even faster deposition rates are possible by adding more modules for additional Al2O3 ALD cycles for every one rotation of the inner cylinder.
Semiautomatic registration of 3D transabdominal ultrasound images for patient repositioning during postprostatectomy radiotherapy
Authors: Presles Benoît , Fargier‐Voiron Marie , Biston Marie‐Claude , Lynch Rod , Munoz Alexandre , Liebgott Hervé , Pommier Pascal , Rit Simon , Sarrut David
Published: December 2014

Purpose:

The aim of the present work is to propose and evaluate registration algorithms of three‐dimensional (3D) transabdominal (TA) ultrasound (US) images to setup postprostatectomy patients during radiation therapy.

Methods:

Three registration methods have been developed and evaluated to register a reference 3D‐TA‐US image acquired during the planning CT session and a 3D‐TA‐US image acquired before each treatment session. The first method (method A) uses only gray value information, whereas the second one (method B) uses only gradient information. The third one (method C) combines both sets of information. All methods restrict the comparison to a region of interest computed from the dilated reference positioning volume drawn on the reference image and use mutual information as a similarity measure. The considered geometric transformations are translations and have been optimized by using the adaptive stochastic gradient descent algorithm. Validation has been carried out using manual registration by three operators of the same set of image pairs as the algorithms. Sixty‐two treatment US images of seven patients irradiated after a prostatectomy have been registered to their corresponding reference US image. The reference registration has been defined as the average of the manual registration values. Registration error has been calculated by subtracting the reference registration from the algorithm result. For each session, the method has been considered a failure if the registration error was above both the interoperator variability of the session and a global threshold of 3.0 mm.

Results:

All proposed registration algorithms have no systematic bias. Method B leads to the best results with mean errors of −0.6, 0.7, and −0.2 mm in left–right (LR), superior–inferior (SI), and anterior–posterior (AP) directions, respectively. With this method, the standard deviations of the mean error are of 1.7, 2.4, and 2.6 mm in LR, SI, and AP directions, respectively. The latter are inferior to the interoperator registration variabilities which are of 2.5, 2.5, and 3.5 mm in LR, SI, and AP directions, respectively. Failures occur in 5%, 18%, and 10% of cases in LR, SI, and AP directions, respectively. 69% of the sessions have no failure.

Conclusions:

Results of the best proposed registration algorithm of 3D‐TA‐US images for postprostatectomy treatment have no bias and are in the same variability range as manual registration. As the algorithm requires a short computation time, it could be used in clinical practice provided that a visual review is performed.
Quantification of stromal vascular cell mechanics with a linear cell monolayer rheometer
Authors: Elkins Claire M. , Shen Wen-Jun , Khor Victor K. , Kraemer Fredric B. , Fuller Gerald G.
Published: December 2014

Over the past few decades researchers have developed a variety of methods for measuring the mechanical properties of whole cells, including traction force microscopy, atomic force microscopy (AFM), and single-cell tensile testing. Though each of these techniques provides insight into cell mechanics, most also involve some nonideal conditions for acquiring live cell data, such as probing only one portion of a cell at a time, or placing the cell in a nonrepresentative geometry during testing. In the present work, we describe the development of a linear cell monolayer rheometer (LCMR) and its application to measure the mechanics of a live, confluent monolayer of stromal vascular cells. In the LCMR, a monolayer of cells is contacted on both top and bottom by two collagen-coated plates and allowed to adhere. The top plate then shears the monolayer by stepping forward to induce a predetermined step strain, while a force transducer attached to the top plate collects stress information. The stress and strain data are then used to determine the maximum relaxation modulus recorded after step-strain, Gr0, referred to as the zero-time relaxation modulus of the cell monolayer. The present study validates the ability of the LCMR to quantify cell mechanics by measuring the change in Gr0 of a confluent cell monolayer upon the selective inhibition of three major cytoskeletal components (actin microfilaments, vimentin intermediate filaments, and microtubules). The LCMR results indicate that both actin- and vimentin-deficient cells had ∼50% lower Gr0 values than wild-type, whereas tubulin deficiency resulted in ∼100% higher Gr0 values. These findings constitute the first use of a cell monolayer rheometer to quantitatively distinguish the roles of different cytoskeletal elements in maintaining cell stiffness and structure. Significantly, they are consistent with results obtained using single-cell mechanical testing methods, suggesting that the rheology-based LCMR technique may be a useful tool for rapid analysis of cell mechanics by shearing an entire cell monolayer.
If Exercise is Medicine, Where is Exercise in Medicine? Review of U.S. Medical Education Curricula for Physical Activity-Related Content
Authors: Cardinal Bradley J. , Park Eugene A. , Kim MooSong , Cardinal Marita K.
Published: December 2014

Background:

This study provides an update on the amount and type of physical activity education occurring in medical education in the United States in 2013. It is the first study to do so since 2002.

Methods:

Applying content analysis methodology, we reviewed all accessible accredited doctor of medicine and doctor of osteopathic medicine institutions’ websites for physical activity education related coursework (N = 118 fully accessible; 69.41%).

Results:

The majority of institutions did not offer any physical activity education–related courses. When offered, they were rarely required. Courses addressing sports medicine and exercise physiology were offered more than courses in other content domains. Most courses were taught using a clinical approach. No differences were observed between MD and DO institutions, or between private and public institutions.

Conclusions:

More than one-half of the physicians trained in the United States in 2013 received no formal education in physical activity and may, therefore, be ill-prepared to assist their patients in a manner consistent with Healthy People 2020, the National Physical Activity Plan, or the Exercise is Medicine initiative. The Bipartisan Policy Center, American College of Sports Medicine, and the Alliance for a Healthier Generation called for a reversal of this situation on June 23, 2014.
Structural analysis of the EGFR/HER3 heterodimer reveals the molecular basis for activating HER3 mutations
Authors: Littlefield Peter , Liu Lijun , Mysore Venkatesh , Shan Yibing , Shaw David E. , Jura Natalia
Published: December 2014

Cancer-associated mutations in the catalytically inactive receptor HER3 enhance its ability to activate other members of the human epidermal growth factor receptor family.
De-Adhesion for Mitotic Progression
Authors: VanHook Annalisa M.
Published: December 2014

The cell cycle–regulated protein DEPDC1B is required for rounding of adherent cells during mitosis.
Stimulatory Signals Add Up for T Cells
Authors: Mueller Kristen L.
Published: December 2014

T cells follow a linear calculation when integrating costimulatory and cytokine signals.
Synergistic Effect of Nitazoxanide with Neuraminidase Inhibitors against Influenza A Viruses
In Vitro
Authors: Belardo Giuseppe , Cenciarelli Orlando , La Frazia Simone , Rossignol Jean Francois , Santoro M. Gabriella
Published: December 2014

The emergence of drug-resistant influenza A virus (IAV) strains represents a serious threat to global human health and underscores the need for novel approaches to anti-influenza chemotherapy. Combination therapy with drugs affecting different IAV targets represents an attractive option for influenza treatment. We have previously shown that the thiazolide anti-infective nitazoxanide (NTZ) inhibits H1N1 IAV replication by selectively blocking viral hemagglutinin maturation. Herein we investigate the anti-influenza activity of NTZ against a wide range of human and avian IAVs (H1N1, H3N2, H5N9, H7N1), including amantadine-resistant and oseltamivir-resistant strains,
in vitro
. We also investigate whether therapy with NTZ in combination with the neuraminidase inhibitors oseltamivir and zanamivir exerts synergistic, additive, or antagonistic antiviral effects against influenza viruses. NTZ was effective against all IAVs tested, with 50% inhibitory concentrations (IC
50
s) ranging from 0.9 to 3.2 μM, and selectivity indexes (SIs) ranging from >50 to >160, depending on the strain and the multiplicity of infection (MOI). Combination therapy studies were performed in cell culture-based assays using A/Puerto Rico/8/1934 (H1N1), A/WSN/1933 (H1N1), or avian A/chicken/Italy/9097/1997 (H5N9) IAVs; dose-effect analysis and synergism/antagonism quantification were performed using isobologram analysis according to the Chou-Talalay method. Combination index (CI) analysis indicated that NTZ and oseltamivir combination treatment was synergistic against A/Puerto Rico/8/1934 (H1N1) and A/WSN/1933 (H1N1) IAVs, with CI values ranging between 0.39 and 0.63, independently of the MOI used. Similar results were obtained when NTZ was administered in combination with zanamivir (CI = 0.3 to 0.48). NTZ-oseltamivir combination treatment was synergistic also against the avian A/chicken/Italy/9097/1997 (H5N9) IAV (CI = 0.18 to 0.31). Taken together, the results suggest that regimens that combine neuraminidase inhibitors and nitazoxanide exert synergistic anti-influenza effects.
Mechanism of Action for Respiratory Syncytial Virus Inhibitor RSV604
Authors: Challa SreeRupa , Scott Andrew D. , Yuzhakov Olga , Zhou Ying , Tiong-Yip Choi Lai , Gao Ning , Thresher Jason , Yu Qin
Published: December 2014

Respiratory syncytial virus (RSV) is the leading cause of acute lower respiratory tract infections in young children and other high-risk populations. RSV nucleoprotein (N) is essential for virus assembly and replication as part of the viral ribonucleoprotein (RNP) complex. RSV604 was a putative N inhibitor in phase 2 clinical trials whose molecular mechanism of action (MoA) was not well understood. This study investigated the cell line-dependent potency of RSV604 and demonstrated its direct binding to the N protein
in vitro
, providing the first evidence of direct target engagement for this class of inhibitors reported to date. The affinity of RSV604 N binding was not affected by RSV604 resistance mutations in the N protein. RSV604 engaged in two different MoAs in HeLa cells, inhibiting both RSV RNA synthesis and the infectivity of released virus. The lack of inhibition of viral RNA synthesis in some cell lines explained the cell-type-dependent potency of the inhibitor. RSV604 did not inhibit viral RNA synthesis in the RSV subgenomic replicon cells or in the cell-free RNP assay, suggesting that it might act prior to viral replication complex formation. RSV604 did not alter N protein localization in the infected cells. Taken together, these results provide new insights leading to an understanding of the MoAs of RSV604 and other similar N inhibitors.
Unexpected Challenges in Treating Multidrug-Resistant Gram-Negative Bacteria: Resistance to Ceftazidime-Avibactam in Archived Isolates of Pseudomonas aeruginosa
Authors: Winkler Marisa L. , Papp-Wallace Krisztina M. , Hujer Andrea M. , Domitrovic T. Nicholas , Hujer Kristine M. , Hurless Kelly N. , Tuohy Marion , Hall Geraldine , Bonomo Robert A.
Published: December 2014

Pseudomonas aeruginosa
is a notoriously difficult-to-treat pathogen that is a common cause of severe nosocomial infections. Investigating a collection of β-lactam-resistant
P. aeruginosa
clinical isolates from a decade ago, we uncovered resistance to ceftazidime-avibactam, a novel β-lactam/β-lactamase inhibitor combination. The isolates were systematically analyzed through a variety of genetic, biochemical, genomic, and microbiological methods to understand how resistance manifests to a unique drug combination that is not yet clinically released. We discovered that avibactam was able to inactivate different AmpC β-lactamase enzymes and that
bla
PDC
regulatory elements and penicillin-binding protein differences did not contribute in a major way to resistance. By using carefully selected combinations of antimicrobial agents, we deduced that the greatest barrier to ceftazidime-avibactam is membrane permeability and drug efflux. To overcome the constellation of resistance determinants, we show that a combination of antimicrobial agents (ceftazidime/avibactam/fosfomycin) targeting multiple cell wall synthetic pathways can restore susceptibility. In
P. aeruginosa
, efflux, as a general mechanism of resistance, may pose the greatest challenge to future antibiotic development. Our unexpected findings create concern that even the development of antimicrobial agents targeted for the treatment of multidrug-resistant bacteria may encounter clinically important resistance. Antibiotic therapy in the future must consider these factors.
Susceptibility Profiles of Amphotericin B and Posaconazole against Clinically Relevant Mucorales Species under Hypoxic Conditions
Authors: Maurer Elisabeth , Binder Ulrike , Sparber Manuela , Lackner Michaela , Caramalho Rita , Lass-Flörl Cornelia
Published: December 2014

The effect of hypoxic conditions on the
in vitro
efficacy of amphotericin B and posaconazole against
Mucorales
was evaluated by defining MICs with Etest and broth microdilution and identifying minimal fungicidal concentrations (MFCs). With Etest, oxygen-dependent changes were detected, while the MIC and the MFC determined with broth microdilution remained unaltered with reduced oxygen levels. The observed differences depended on the method used.
Substrate-Induced Radical Formation in 4-Hydroxybutyryl Coenzyme A Dehydratase from Clostridium aminobutyricum
Authors: Zhang Jin , Friedrich Peter , Pierik Antonio J. , Martins Berta M. , Buckel Wolfgang
Published: December 2014

4-Hydroxybutyryl-coenzyme A (CoA) dehydratase (4HBD) from
Clostridium aminobutyricum
catalyzes the reversible dehydration of 4-hydroxybutyryl-CoA to crotonyl-CoA and the irreversible isomerization of vinylacetyl-CoA to crotonyl-CoA. 4HBD is an oxygen-sensitive homotetrameric enzyme with one [4Fe-4S]
2+
cluster and one flavin adenine dinucleotide (FAD) in each subunit. Upon the addition of crotonyl-CoA or the analogues butyryl-CoA, acetyl-CoA, and CoA, UV-visible light and electron paramagnetic resonance (EPR) spectroscopy revealed an internal one-electron transfer to FAD and the [4Fe-4S]
2+
cluster prior to hydration. We describe an active recombinant 4HBD and variants produced in
Escherichia coli
. The variants of the cluster ligands (H292C [histidine at position 292 is replaced by cysteine], H292E, C99A, C103A, and C299A) had no measurable dehydratase activity and were composed of monomers, dimers, and tetramers. Variants of other potential catalytic residues were composed only of tetramers and exhibited either no measurable (E257Q, E455Q, and Y296W) hydratase activity or <1% (Y296F and T190V) dehydratase activity. The E455Q variant but not the Y296F or E257Q variant displayed the same spectral changes as the wild-type enzyme after the addition of crotonyl-CoA but at a much lower rate. The results suggest that upon the addition of a substrate, Y296 is deprotonated by E455 and reduces FAD to FADH·, aided by protonation from E257 via T190. In contrast to FADH·, the tyrosyl radical could not be detected by EPR spectroscopy. FADH· appears to initiate the radical dehydration via an allylic ketyl radical that was proposed 19 years ago. The mode of radical generation in 4HBD is without precedent in anaerobic radical chemistry. It differs largely from that in enzymes, which use coenzyme B
12
,
S
-adenosylmethionine, ATP-driven electron transfer, or flavin-based electron bifurcation for this purpose.
Recombinant Protein Truncation Strategy for Inducing Bactericidal Antibodies to the Macrophage Infectivity Potentiator Protein of Neisseria meningitidis and Circumventing Potential Cross-Reactivity with Human FK506-Binding Proteins
Authors: Bielecka Magdalena K. , Devos Nathalie , Gilbert Mélanie , Hung Miao-Chiu , Weynants Vincent , Heckels John E. , Christodoulides Myron
Published: December 2014

A recombinant macrophage infectivity potentiator (rMIP) protein of
Neisseria meningitidis
induces significant serum bactericidal antibody production in mice and is a candidate meningococcal vaccine antigen. However, bioinformatics analysis of MIP showed some amino acid sequence similarity to human FK506-binding proteins (FKBPs) in residues 166 to 252 located in the globular domain of the protein. To circumvent the potential concern over generating antibodies that could recognize human proteins, we immunized mice with recombinant truncated type I rMIP proteins that lacked the globular domain and the signal leader peptide (LP) signal sequence (amino acids 1 to 22) and contained the His purification tag at either the N or C terminus (C-term). The immunogenicity of truncated rMIP proteins was compared to that of full (i.e., full-length) rMIP proteins (containing the globular domain) with either an N- or C-terminal His tag and with or without the LP sequence. By comparing the functional murine antibody responses to these various constructs, we determined that C-term His truncated rMIP (−LP) delivered in liposomes induced high levels of antibodies that bound to the surface of wild-type but not Δ
mip
mutant meningococci and showed bactericidal activity against homologous type I MIP (median titers of 128 to 256) and heterologous type II and III (median titers of 256 to 512) strains, thereby providing at least 82% serogroup B strain coverage. In contrast, in constructs lacking the LP, placement of the His tag at the N terminus appeared to abrogate bactericidal activity. The strategy used in this study would obviate any potential concerns regarding the use of MIP antigens for inclusion in bacterial vaccines.
CD73-Generated Adenosine Is Critical for Immune Regulation during Toxoplasma gondii Infection
Authors: Mahamed Deeqa A. , Toussaint Leon E. , Bynoe Margaret S.
Published: December 2014

As an obligate intracellular pathogen, the apicomplexan parasiteToxoplasma gondiievades immune system-mediated clearance by undergoing stage differentiation to persist indefinitely in susceptible hosts. Previously, we found that mice deficient in the ectoenzyme CD73, which generates adenosine in the extracellular matrix, were resistant to chronic toxoplasmosis after oral infection withT. gondii. Resistance in CD73 knockout mice was due to a delay in parasite differentiation in the central nervous system (CNS). To further clarify the role of CD73 and extracellular adenosine inT. gondiipathogenesis, we infected wild-type (WT) and CD73−/−mice withT. gondiicysts systemically by the intraperitoneal (i.p.) route. In contrast to oral infection, i.p. infected CD73−/−mice were highly susceptible to immune-mediated pathology, with significantly increased infiltration of neutrophils and T cells into the peritoneal cavity. Administration of the broad-spectrum adenosine receptor agonist 5′-N-ethylcarboxamidoadenosine (NECA) protected CD73−/−mice againstT. gondii-induced immunopathology, suggesting that the absence of CD73-generated adenosine led to the increased susceptibility in these mice. Peritoneal exudate cells from infected CD73−/−mice produced higher levels of the inflammatory mediators nitric oxide, tumor necrosis factor alpha (TNF-α), and interleukin-1β (IL-1β), without enhanced parasite killing or clearance. Bone marrow chimeras established that CD73 expression in both hematopoietic and nonhematopoietic compartments contributes to limitingT. gondii-induced immunopathology. In addition, mice deficient in the adenosine receptor A2Awere more susceptible to immunopathology during intraperitoneal infection withT. gondiithan WT mice. Thus, extracellular adenosine is a key immune regulator that limits collateral tissue damage due to an intracellular pathogen and promotes host survival.
Fifty Ways To Inhibit Motility via Cyclic Di-GMP: the Emerging Pseudomonas aeruginosa Swarming Story
Authors: McCarter Linda L. , Gomelsky Mark
Published: December 2014

There are numerous ways by which cyclic dimeric GMP (c-di-GMP) inhibits motility. Kuchma et al. (S. L. Kuchma, N. J. Delalez, L. M. Filkins, E. A. Snavely, J. P. Armitage, and G. A. O'Toole, J. Bacteriol. 197:420–430, 2015,http://dx.doi.org/10.1128/JB.02130-14) offer a new, previously unseen way of swarming motility inhibition inPseudomonas aeruginosaPA14. This bacterium possesses a single flagellum with one rotor and two sets of stators, only one of which can provide torque for swarming. The researchers discovered that elevated levels of c-di-GMP inhibit swarming by skewing stator selection in favor of the nonfunctional, “bad” stators.
Different Effects of Transcutaneous Electric Nerve Stimulation and Electroacupuncture at St36–st37 on the Cerebral Cortex
Authors: Kang Yu-Tien , Liao Yi-Sheng , Hsieh Ching-Liang
Published: November 2014

Background

The effects of transcutaneous electric nerve stimulation (TENS) and electroacupuncture (EA) on the cerebral cortex are largely unclear. The purpose of the present study was to investigate the effect of TENS and EA on the cerebral cortex by examining their effect on the median nerve-somatosensory evoked potentials (MN-SEPs).

Methods

Twenty volunteers were studied. The cortical and cervical spinal potentials were recorded by median nerve stimulation at the left wrist. Sham TENS, 2 Hz TENS and 2 Hz EA were applied to both ST36 and ST37. MN-SEPs were recorded during sham TENS, 2 Hz TENS and 2 Hz EA, with at least 1 week interval for each subject. One-way analysis of variance was used to determine the differences in latency and amplitude of the MN-SEPs observed in the stimulation and post-stimulation periods compared with baseline. Scheffe's post hoc correction was employed to identify pairwise differences.

Results

No differences in mean latency were found between the stimulation procedures during the stimulation and post-stimulation periods. 2 Hz EA but not sham TENS or 2 Hz TENS caused higher mean amplitudes in N20 and N30 during the stimulation and post-stimulation periods.

Conclusions

EA, but not TENS, induces changes in certain components of the signal.