Πέμπτη 6 Σεπτεμβρίου 2018

Structural Variants and Selective Sweep Foci Contribute to Insecticide Resistance in the Drosophila Genetic Reference Panel

Patterns of nucleotide polymorphism within populations of Drosophila melanogaster suggest that insecticides have been the selective agents driving the strongest recent bouts of positive selection. However, there is a need to explicitly link selective sweeps to the particular insecticide phenotypes that could plausibly account for the drastic selective responses that are observed in these non-target insects. Here, we screen the Drosophila Genetic Reference Panel with two common insecticides; malathion (an organophosphate) and permethrin (a pyrethroid). Genome-wide association studies map survival on malathion to the two of the largest sweeps in the D. melanogaster genome; Ace and Cyp6g1. Malathion survivorship also correlates with lines which have high levels of Cyp12d1, Jheh1 and Jheh2 transcript abundance. Permethrin phenotypes map to the largest cluster of P450 genes in the Drosophila genome, however in contrast to a selective sweep driven by insecticide use, the derived allele seems to be associated with susceptibility. These results underscore previous findings that highlight the importance of structural variation to insecticide phenotypes: Cyp6g1 exhibits copy number variation and transposable element insertions, Cyp12d1 is tandemly duplicated, the Jheh loci are associated with a Bari1 transposable element insertion, and a Cyp6a17 deletion is associated with susceptibility



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Signatures of Insecticide Selection in the Genome of Drosophila melanogaster

Resistance to insecticides has evolved in multiple insect species, leading to increased application rates and even control failures. Understanding the genetic basis of insecticide resistance is fundamental for mitigating its impact on crop production and disease control. We performed a GWAS approach with the Drosophila Genetic Reference Panel (DGRP) to identify the mutations involved in resistance to two widely used classes of insecticides: organophosphates (OPs, parathion) and pyrethroids (deltamethrin). Most variation in parathion resistance was associated with mutations in the target gene Ace, while most variation in deltamethrin resistance was associated with mutations in Cyp6a23, a gene encoding a detoxification enzyme never previously associated with resistance. A "nested GWAS" further revealed the contribution of other loci: Dscam1 and trpl were implicated in resistance to parathion, but only in lines lacking Wolbachia. Cyp6a17, the paralogous gene of Cyp6a23, and CG7627, an ATP-binding cassette transporter, were implicated in deltamethrin resistance. We observed signatures of recent selective sweeps at all of these resistance loci and confirmed that the soft sweep at Ace is indeed driven by the identified resistance mutations. Analysis of allele frequencies in additional population samples revealed that most resistance mutations are segregating across the globe, but that frequencies can vary substantially among populations. Altogether, our data reveal that the widely used OP and pyrethroid insecticides imposed a strong selection pressure on natural insect populations. However, it remains unclear why, in Drosophila, resistance evolved due to changes in the target site for OPs, but due to a detoxification enzyme for pyrethroids.



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Frequency and circumstances of falls reported by ambulatory unilateral lower limb prosthesis users: a secondary analysis

More than 50% of lower limb prosthesis (LLP) users report falling at least once a year, placing them at high risk for adverse health outcomes like reduced mobility and diminished quality-of-life. Efforts to reduce falls among LLP users have traditionally focused on developing clinical tests to assess fall risk, designing prosthetic components to improve patient safety, and identifying risk factors to recognize potential fallers. Little attention has been directed towards recording, reporting, and characterizing the circumstances of falls among LLP users.

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Bone Mineral Density among Individuals with Residual Lower Limb Weakness after Polio

Literature indicates that individuals with long-term residual lower extremity (LE) weakness after polio have decreased bone mineral density (BMD) deficiencies related to muscle weakness. Where weakness is asymmetrical, bone densitometry (BDt) measured only on the stronger LE may misclassify BMD.

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Ankle joint and rearfoot biomechanics during toe-in and toe-out walking in people with medial compartment knee osteoarthritis

Toe-in and toe-out walking are two strategies which have been shown to be effective in reducing the knee adduction moment in people with knee osteoarthritis. However, despite a positive biomechanical impact on the knee, altering foot rotation may impart unintended forces or joint positions on the ankle that could impact joint health. The kinematic and kinetic changes at the ankle during toe-in or toe-out walking have yet to be examined.

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A Descriptive Study of Self-Reported Injury in Non-elite Adaptive Athletes

Adaptive sports programs are increasing across the country and there is a paucity of research investigating the epidemiology and sports injury risk factors in non-elite athletes.

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Treatment of Knee Meniscus Pathology: Rehabilitation, Surgery, and Orthobiologics

The meniscal tear treatment paradigm traditionally begins with conservative measures such as physical therapy with referral for operative management for persistent or mechanical symptoms. As a result, the partial meniscectomy is performed more than any other orthopedic procedure in the United States. This treatment paradigm has shifted as recent literature supports the attempt to preserve or repair the meniscus whenever possible given its importance for the structural integrity of the knee joint and the risk of early osteoarthritis associated after meniscus excision.

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Is there an impact of cervical plating on the development of adjacent segment degeneration following Smith-Robinson procedure ? A magnetic resonance imaging study of 84 patients with a 24 year follow-up

Anterior cervical discectomy and fusion (ACDF) without and with cervical plating (ACDF+CP) are accepted surgical techniques for the treatment of degenerative cervical disc disorders. The effect of CP on the development of adjacent segment degeneration (ASD) remains unclear.

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Polaris® expands its public safety line to include all-electric GEM® vehicles

MINNEAPOLIS — Polaris® Government and Defense is adding all-electric GEM® vehicles to its line of highly mobile, affordable, and professional public safety vehicles. Polaris firefighting, law enforcement, and rescue RANGER® and GENERAL® vehicles are proven effective and can go places cars, trucks, helicopters, and other alternatives can't. Now,...

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Periodic Limb Movements in Sleep: Prevalence and Associated Sleepiness in the Wisconsin Sleep Cohort

Periodic limb movements in sleep (PLMS)1 are repetitive bursts of muscle activity, typically in the lower limbs, which may be accompanied by an arousal or sleep fragmentation(Pollmacher et al., 1993). Although PLMS are believed to increase with age(Ancoli-Israel et al., 1985, Ferri et al., 2008), until recently, the prevalence of PLMS in the general population had not been reliably estimated. Small studies have reported prevalence rates between 5 and 11%(Hornyak et al., 2006, Scofield et al., 2008), including a community-based sample of 592 participants recruited from the general population of tri-country Detroit which reported 7.6% (Hornyak et al., 2006, Scofield et al., 2008).

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EEG coherence in a mental arithmetic task performance in first episode schizophrenia and schizoaffective disorder

Current theories of schizophrenia (SZ) have emphasized that the pathophysiology arises from a 'dysconnectivity' between and within cortical areas of the brain (Friston, 1999, Uhlhaas and Singer, 2010, Stephan et al., 2009). Studies of functional connectivity in patients with schizophrenia using PET and functional MRI data both during cognitive tasks (Potkin and Ford, 2009 for a review) and at rest (for a review see Greicius, 2008) vastly support this hypothesis. However, less attention has been paid to the particular disorders within the schizophrenia spectrum.

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The utility and application of electrophysiological methods in the study of visual hallucinations

Visual hallucinations (VH) are a common symptom in both clinical and non-clinical populations, including a wide range of psychiatric, neurological, and ophthalmologic conditions such as schizophrenia, neurodegenerative dementias, eye disease, delirium and drug-induced hallucinosis (Abraham and Duffy 1996; Menon et al. 2003; Collerton et al. 2005; Armstrong (2012); ffytche and Wible 2014).

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Atypical perceptual and neural processing of emotional prosodic changes in children with Autism Spectrum Disorders

Deficient social communication skills, narrow interests, and repetitive behavior are the main diagnostic features of autism spectrum disorders (ASD) (APA, 2013). Some individuals with ASD show significant delays and abnormalities in their language development (later referred to as children with ASD (LI) in this article), whereas some individuals with ASD show rather typical formal language development (later referred to as children with ASD (no LI)) (WHO, 1993; Rapin and Dunn, 2003; Gillberg and Coleman, 2000).

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Pulsara adds key communication features to its healthcare mobile platform

Pulsara is proud to announce the release of two new ingenious elements of its application, Video Chat and Audio Clips, that will enable clinicians to communicate in a much broader way. 2018 has shown to be a breakthrough year for this company with a number of major feature releases, including the recent Consult/Transfer component. These features further enhance regional communication amongst...

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Does the King Airway system signal the end of intubation skills?

Our co-hosts tackle the topic of supraglottic airways, and the best way to approach them in the field

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Does the King Airway system signal the end of intubation skills?

Our co-hosts tackle the topic of supraglottic airways, and the best way to approach them in the field

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Tackling Loneliness

Most of us would not consider loneliness a public health epidemic, yet startling new findings show that a majority of Americans are lonely – according to a survey that Cigna conducted with 20,000 Americans – and loneliness is at the heart of physical and mental health. Not only did our new study find that most Americans are considered lonely, but those who are lonely are much more...

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Decontamination of select infrastructure materials after a radiological incident using a water-based formulation

Publication date: October 2018

Source: Journal of Environmental Radioactivity, Volumes 193–194

Author(s): Jaleh Semmler, Anthony Snaglewski, Arshad Toor, Zahid Khan, Wenxing Kuang, Konstantin Volchek, Pervez Azmi, Vladimir Blinov, Carl Brown

Abstract

This paper summarizes the results of the decontamination of the infrastructure materials concrete, limestone, brick and asphalt contaminated with 60Co, 85Sr, 137Cs and 241Am. The paper focuses on the effect of differences in substrate properties and of the pH of the radionuclide solution used for surface contamination on adsorption or ion exchange of the radionuclides and how these factors affect the decontamination effectiveness. A six-component chemical formulation was used and a process effectiveness of up to 76% was obtained depending on the substrate and radionuclide. Asphalt was the easiest material to decontaminate because of its more hydrophobic nature. Concrete and limestone (and to some extent brick) were less effectively decontaminated as their porous surfaces allowed penetration of radionuclides into water-filled pores in the substrate facilitating adsorption or ion exchange and making them difficult to remove. Brick was the most difficult material to decontaminate because the major component of brick is clay which retains most mono- and divalent ions. The removal of 60Co, 85Sr and 137Cs from the surfaces of concrete, limestone and brick increased when the pH of the radionuclide solutions was moderately acidic to neutral compared to when they were highly acidic. The variability in the test results was similar to that observed in other studies using other decontamination methods, attributed to the inhomogeneity of the substrates used and considered representative of real infrastructure materials.



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Corrigendum to “Dating the Middle Palaeolithic deposits of La Quina Amont (Charente, France) using luminescence methods” [Journal of Human Evolution 109 (2017) 30–45]

Publication date: Available online 6 September 2018

Source: Journal of Human Evolution

Author(s): Marine Frouin, Christelle Lahaye, Hélène Valladas, Thomas Higham, Anne Delagnes, Norbert Mercier



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Psychophysiological and self-reported responses in individuals with methamphetamine use disorder exposed to emotional video stimuli

Publication date: Available online 6 September 2018

Source: International Journal of Psychophysiology

Author(s): Xi-Jing Chen, Chun-Guang Wang, Yong-Hui Li, Nan Sui

Abstract

Individuals with methamphetamine use disorder (MUD) exhibit irritability and compulsive emotional responses, yet the relevant study is scarce. The characteristic of their positive and negative emotional responses can provide effective targets for the clinical intervention. In this study, we compared the emotional responses of 60 participants with MUD and 30 healthy participants to visual stimuli. They watched four types of video to elicit anger, fear, amusement, and joy emotional responses. The self-report of emotional responses (i.e., arousal, valence, and proximity), skin conductance level, and startle response were measured. Comparing to the healthy controls, the methamphetamine group's subjective arousal level of fear is significantly lower (t = 3.763, p < .01); the skin conductance level of joy is significantly higher (t = 2.086, p < .05), and the level of anger is marginal significantly higher (t = 1.984, p = .05); the startle response level of anger (t = 2.069, p < .05) and joy (t = 2.406, p < .05) is significantly higher. The methamphetamine group exhibited an enhanced emotion response to anger and a decreased response to joy which may indicate the emotion dysregulation problem caused by drug. These results provide effective targets for clinical intervention in treating patients of MUD with emotion dysregulation problems.



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The utility and application of electrophysiological methods in the study of visual hallucinations

Publication date: Available online 6 September 2018

Source: Clinical Neurophysiology

Author(s): Katrina daSilva Morgan, Greg J. Elder, Dominic H. ffytche, Daniel Collerton, John-Paul Taylor

Abstract

Visual hallucinations (VH) are a common symptom in multiple clinical and non-clinical populations. Although structural and functional neuroimaging has informed the understanding of VH, temporal resolution is limited. Electrophysiological techniques provide a complimentary perspective on dynamic and temporal aspects of neural functioning, offering greater insight into the mechanisms underlying their formation. In this review we examine and critically evaluate the emerging evidence base utilising electrophysiological approaches in the study of VH. Overall, increased visual system excitability, dysfunctional visual processing and network connectivity, and cholinergic dysfunction have been consistently observed in VH-prone pathologies. However, a major limitation is in the lack of robust experimental studies and the reliance on single case reports. We conclude that electrophysiology provides tentative evidence for the contribution of bottom-up, top-down, and network dysfunction in the aetiology of VH, supporting several existing VH models. Furthermore, we discuss how electrophysiology has been directly utilised in specific clinical interventions for VH. Further exploration utilising electrophysiology in combination with, for example, neuroimaging will help better understand VH aetiology while aiding in the development of novel therapeutic interventions for this difficult to treat symptom.



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Atypical perceptual and neural processing of emotional prosodic changes in children with Autism Spectrum Disorders

Publication date: Available online 6 September 2018

Source: Clinical Neurophysiology

Author(s): R. Lindström, T. Lepistö-Paisley, T. Makkonen, O. Reinvall, T. Nieminen-von Wendt, R. Alén, T. Kujala

Abstract
Objective

The present study explored the processing of emotional speech prosody in school-aged children with autism spectrum disorders (ASD) but without marked language impairments (children with ASD [no LI]).

Methods

The mismatch negativity (MMN)/the late discriminative negativity (LDN), reflecting pre-attentive auditory discrimination processes, and the P3a, indexing involuntary orienting to attention-catching changes, were recorded to natural word stimuli uttered with different emotional connotations (neutral, sad, scornful and commanding). Perceptual prosody discrimination was addressed with a behavioral sound-discrimination test.

Results

Overall, children with ASD (no LI) were slower in behaviorally discriminating prosodic features of speech stimuli than typically developed control children. Further, smaller standard-stimulus event related potentials (ERPs) and MMN/LDNs were found in children with ASD (no LI) than in controls. In addition, the amplitude of the P3a was diminished and differentially distributed on the scalp in children with ASD (no LI) than in control children.

Conclusions

Processing of words and changes in emotional speech prosody is impaired at various levels of information processing in school-aged children with ASD (no LI).

Significance

The results suggest that low-level speech sound discrimination and orienting deficits might contribute to emotional speech prosody processing impairments observed in ASD.



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The cerebellum seems not a ‘little brain’ for the autonomic nervous system

Publication date: Available online 6 September 2018

Source: Clinical Neurophysiology

Author(s): Ryuji Sakakibara



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Cerebellar impairment during an orthostatic challenge in patients with neurogenic orthostatic hypotension

Publication date: Available online 6 September 2018

Source: Clinical Neurophysiology

Author(s): Jacquie Baker, Justin R. Paturel, Kurt Kimpinski

Abstract
Objective

Compare activation patterns within the cortical autonomic network in patients with neurogenic orthostatic hypotension (NOH) versus healthy age-matched controls during an orthostatic challenge.

Methods

Fifteen health controls and 15 NOH patients performed 3 Valsalva maneuvers, and 5-minutes of lower-body negative pressure (LBNP) during a functional brain MRI.

Results

Compared to controls, NOH patients had significantly less activation within the cerebellum during both LBNP and VM. Both groups had significant activation of the bilateral insula and left thalamus during LBNP. No significant differences were found during the recovery phase of LBNP.

Conclusions

The cerebellum, which plays an important role in vestibulo-sympathetic reflexes, important for blood pressure adjustments during postural changes, appear to be affected in patients with NOH. The cerebellum also appears to be affected during other baroreflex mediated stressors such as the VM.

Significance

Orthostatic reflexes mediated by the cerebellum may be impaired in patients with NOH. The results suggest an additional pathological pathway in patients with autonomic failure.



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Silybin Alleviates Hepatic Steatosis and Fibrosis in NASH Mice by Inhibiting Oxidative Stress and Involvement with the Nf-κB Pathway

Abstract

Background and Aim

Silybin is the major biologically active compound of silymarin, the standardized extract of the milk thistle (Silybum marianum). Increasing numbers of studies have shown that silybin can improve nonalcoholic steatohepatitis (NASH) in animal models and patients; however, the mechanisms underlying silybin's actions remain unclear.

Methods

Male C57BL/6 mice were fed a methionine-choline deficient (MCD) diet for 8 weeks to induce the NASH model, and silybin was orally administered to the NASH mice. The effects of silybin on lipid accumulation, hepatic fibrosis, oxidative stress, inflammation-related gene expression and nuclear factor kappa B (NF-κB) activities were evaluated by biochemical analysis, immunohistochemistry, immunofluorescence, quantitative real-time PCR and western blot.

Results

Silybin treatment significantly alleviated hepatic steatosis, fibrosis and inflammation in MCD-induced NASH mice. Moreover, silybin inhibited HSC activation and hepatic apoptosis and prevented the formation of MDBs in the NASH liver. Additionally, silybin partly reversed the abnormal expression of lipid metabolism-related genes in NASH. Further study showed that the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway played important roles in the silybin-derived antioxidant effect, as evidenced by the upregulation of Nrf2 target genes in the silybin treatment group. In addition, silybin significantly downregulated the expression of inflammation-related genes and suppressed the activity of NF-κB signaling.

Conclusions

Silybin was effective in preventing the MCD-induced increases in hepatic steatosis, fibrosis and inflammation. The effect was related to alteration of lipid metabolism-related gene expression, activation of the Nrf2 pathway and inhibition of the NF-κB signaling pathway in the NASH liver.



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Homocysteine and Folate in Inflammatory Bowel Disease: Can Reducing Sulfur Reduce Suffering?



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Blood pressure-independent increase in the cortical cerebral blood flow induced by manual acupuncture of the auricular region in rats

Abstract

We aimed to determine whether acupuncture to the auricular region increases cortical regional cerebral blood flow (rCBF). The rCBF was measured using laser speckle contrast imaging in urethane-anesthetized rats. Acupuncture stimulation was performed manually at the auricular concha or abdomen. The former's stimulation significantly increased the rCBF of the bilateral cerebral cortex in the frontal, parietal, and occipital lobes without altering the systemic arterial pressure. In contrast, abdominal stimulation affected neither rCBF nor systemic arterial pressure. The increase in the rCBF was completely abolished by the severance of the somatic nerves that innervated the auricular region, comprising the trigeminal nerve, facial nerve, auricular branch of the vagal nerve, glossopharyngeal nerve, and great auricular nerve. Thus, application of acupuncture to the auricular region increases the rCBF without increasing arterial pressure.



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Correction to: Differences in gut microbiota associated with age, sex, and stool consistency in healthy Japanese subjects

The authors would like to correct the errors in the publication of the original article. The correction details are given below for your reading.



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Association between physician burnout and patient safety, professionalism, and patient satisfaction: A systematic review and meta-analysis

JAMA Internal Medicine

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Genotype and phenotype analysis using an epilepsy‐associated gene panel in Chinese pediatric epilepsy patients

Clinical Genetics, Volume 0, Issue ja, -Not available-.


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Cultural equivalence, reliability and utility of the Portuguese version of the Oral Behaviors Checklist

Journal of Oral Rehabilitation, Volume 0, Issue ja, -Not available-.


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Pharmacogenetic and clinical predictors of response to clopidogrel plus aspirin after acute coronary syndrome in Egyptians

Objectives Dual antiplatelet therapy (DAPT) with aspirin and clopidogrel reduces the risk for recurrent cardiovascular events after acute coronary syndrome (ACS). However, there is significant variation in response to DAPT that may be influenced by both genetic and nongenetic factors. This study aimed to assess the effect of genetic polymorphisms in PON-1, PEAR-1, P2Y12, CES1, and CYP2C19, along with clinical, demographic, and social factors, on variation in response to DAPT in Egyptians. Participants and methods This study included 230 Egyptians treated with clopidogrel 75 mg/day and aspirin 81 mg/day for at least 12 months following their first ACS. Simple and multivariable logistic regression analyses were carried out to identify factors associated with major adverse cardiovascular events (MACE), defined as the occurrence of recurrent ACS, ischemic stroke, stent-related revascularization, or death, in clopidogrel-treated participants. Results Using multivariable logistic regression analysis, the CYP2C19*2 polymorphism was the only genetic predictor of MACE [odds ratio (OR): 2.23, 95% confidence interval (CI): 1.15–4.33, P=0.01]. In addition, proton pump inhibitor use (OR: 4.77, 95% CI: 1.47–15.54, P=0.009) and diabetes (OR: 1.83, 95% CI: 1.03–3.26, P=0.03) were associated with higher cardiovascular risk, whereas statin use was associated with lower risk (OR: 0.43, 95% CI: 0.25–0.76, P=0.003). The contribution of these four genetic and nongenetic factors explained 19% of the variability in risk for MACE in Egyptians treated with DAPT. Conclusions These results highlight that CYP2C19*2, along with diabetes, and use of proton pump inhibitor and statin are important factors jointly associated with variability in clinical response to DAPT following ACS in Egyptians. *Shaimaa Fathy and Mohamed H. Shahin contributed equally to the writing of this article. Correspondence to Mohamed H. Shahin, PhD, Department of Pharmacotherapy and Translational Research, Center for Pharmacogenomics, College of Pharmacy, University of Florida, PO Box 100486, Gainesville, FL 32610, USA Tel: +1 352 273 6405; fax: +1 352 273 6121; e-mail: mhossam@ufl.edu Received April 18, 2018 Accepted August 17, 2018 Copyright © 2018 Wolters Kluwer Health, Inc. All rights reserved.

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Physiological adaptations to resistance training in rats selectively bred for low and high response to aerobic exercise training

Experimental Physiology, Volume 0, Issue ja, -Not available-.


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Effect of negative motivation on the behavioral and autonomic correlates of deception

Psychophysiology, EarlyView.


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Impact of vascular disease, amputation level, and the mismatch between balance ability and balance confidence in a cross sectional study of the likelihood of falls among people with limb loss: perception versus reality

Objective Investigate impact of balance ability, Activities-specific Balance Confidence (ABC), and other self-reported and clinical factors on incidence of falls among people with lower-limb loss. Design Cross-sectional study (N=305) with multivariable logistic regression analysis Results Participants included 68.3% males with age 55.5±14.9; with 50% dysvascular and 56.8% transtibial amputations. Average ABC=2.1/4 ±1.1, balance ability=2.9/4 ±1.3 and walking speed=0.766±0.387 m/s. The final model showed fall risk was heightened for people with vascular comorbidities (odds ratio [OR]=3.46, 95% confidence interval [CI]=1.40-8.54) and better balance (OR=23.29, CI=3.19-170.23); but attenuated for people with transfemoral (OR=0.08, CI=0.01-0.82) and vascular amputations (OR=0.38, CI=0.15-0.95). Significant interactions existed between age and amputation level (OR=1.06, CI=1.02-1.11) and between balance confidence and balance ability (OR=0.27, CI=0.13-0.57). Conclusions While people with vascular amputations were less likely to fall than those with nonvascular amputations, people with concurrent vascular comorbidities were more likely to fall than those without. People with transfemoral amputations were less likely to fall—however, fall risk increased with each year of age compared to people with transtibial amputations. People with balance ability ≥3.5 fell more often than those with lower ability, but people with lower balance ability and mismatched confidence in their balance ability had 3.7 times greater fall risk. CORRESPONDING AUTHOR CONTACT INFORMATION: Christopher Kevin Wong, PT, PhD, OCS, Associate Professor, Department of Rehabilitation and Regenerative Medicine, Associate Director, Program in Physical Therapy, Columbia University Medical Center, 617 West 168th Street, The Georgian 3rd Floor, New York, NY 10032, Tel: 212-305-3781 Fax: 212-305-4569 Email: ckw7@cumc.columbia.edu DISCLOSURES: a. The authors have no conflicts of interest to report. c. Funding: Supported in part by the National Center for Injury Prevention and Control, Centers for Disease Control and Prevention (grant no. 1 R49 CE002096). Its contents are solely the responsibility of the authors and do not represent the official views of the Centers for Disease Control and Prevention or the Department of Health and Human Services. c. The authors have no potential financial benefits to report. d. Prior presentations: This work has not been presented. Related work focused on injury was presented at the International Society of Prosthetics and Orthotics, 2017 World Conference, Capetown, South Africa. Copyright © 2018 Wolters Kluwer Health, Inc. All rights reserved.

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