Molecular mechanism of Aurora A kinase autophosphorylation and its allosteric activation by TPX2.
We elucidate the molecular mechanisms of two distinct activation strategies (autophosphorylation and TPX2-mediated activation) in human Aurora A kinase. Classic allosteric activation is in play where either activation loop phosphorylation or TPX2 binding to a conserved hydrophobic groove shifts the equilibrium far towards the active conformation. We resolve the controversy about the mechanism of autophosphorylation by demonstrating intermolecular autophosphorylation in a long-lived dimer by combining X-ray crystallography with functional assays. We then address the allosteric activation by TPX2 through activity assays and the crystal structure of a domain-swapped dimer of dephosphorylated Aurora A and TPX2(1-25). While autophosphorylation is the key regulatory mechanism in the centrosomes in the early stages of mitosis, allosteric activation by TPX2 of dephosphorylated Aurora A could be at play in the spindle microtubules. The mechanistic insights into autophosphorylation and allosteric activation by TPX2 binding proposed here, may have implications for understanding regulation of other protein kinases.DOI: http://dx.doi.org/10.7554/eLife.02667.001.


Biochemically normal adrenal pheochromocytoma following extensive central necrosis in a child with von Hippel-Lindau (VHL) gene mutation.
Pheochromocytomas are rare in children. The diagnosis is usually established from a raised urinary or plasma catecholamine or their metabolites. We present a girl aged 11 years who manifested with a hypertensive crisis secondary to an adrenal tumour but with unexpectedly normal urinary metanephrine and catecholamine results. She improved spontaneously following the crisis and underwent surgery later. The histopathological study confirmed a pheochromocytoma with large central necrosis. Her genetic screening reported a pathogenic von Hippel-Lindau gene mutation. Surveillance scan postsurgery detected no other tumours. Following the catecholamine crisis, an acute infarct occurred, resulting in extensive tumour necrosis and subsequent rapid remission of symptoms and paradoxically normal biochemical markers. Although not unheard of in adults, we believe this is the first reported case of an extensive spontaneous necrosis resulting in a biochemically normal pheochromocytoma in a child.


Internet abusers associate with a depressive state but not a depressive trait.
The present study investigated three issues: (i) whether Internet abusers display a depressive state without a depressive trait; (ii) which symptoms are shared between Internet abuse and depression; and (iii) which personality characteristics were shown in Internet abusers. Ninety-nine male and 58 female participants aged 18-24 years were screened with the Chen Internet Addiction Scale. After screening, subjects were separated into the high- (n = 73) and low-risk (n = 84) Internet abuser groups. Participants were respectively administered the Chinese version of the Beck Depression Inventory-II to assess a depressive state and the Minnesota Multiphasic Personality Inventory-2 to assess a depressive trait. The present results showed that high-risk Internet abusers exhibited a stronger depressive state than low-risk Internet abusers in the Beck Depression Inventory-II. However, high-risk Internet abusers didnot show a depressive trait in the Minnesota Multiphasic Personality Inventory-2 compared to low-risk Internet abusers. Therefore, high-risk Internet abuse participants exhibited a depressive state without a depressive trait. In a comparison of the symptoms of depression and Internet abuse, it was found that high-risk Internet abuse participants shared some common behavioral mechanisms with depression, including the psychiatric symptoms of loss of interest, aggressive behavior, depressive mood, and guilty feelings. High-risk Internet abuse participants may be more susceptible to a temporal depressive state but not a permanent depressive trait. The present findings have clinical implications for the prevention and treatment of Internet abuse.


High-Throughput CRISPR/Cas9 Mutagenesis Streamlines Trait Gene Identification in Maize.
Maize (Zea mays) is one of the most important crops in the world. However, few agronomically important maize genes have been cloned and used for trait improvement, due to its complex genome and genetic architecture. Here, we integrated multiplexed CRISPR/Cas9-based high-throughput targeted mutagenesis with genetic mapping and genomic approaches to successfully target 743 candidate genes corresponding to traits relevant for agronomy and nutrition. After low-cost barcode-based deep sequencing, 412 edited sequences covering 118 genes were precisely identified from individuals showing clear phenotypic changes. The profiles of the associated gene-editing events were similar to those identified in human cell lines and consequently are predictable using an existing algorithm originally designed for human studies. We observed unexpected but frequent homology-directed repair through endogenous templates that was likely caused by spatial contact between distinct chromosomes. Based on the characterization and interpretation of gene function from several examples, we demonstrate that the integration of forward and reverse genetics via a targeted mutagenesis library promises rapid validation of important agronomic genes for crops with complex genomes. Beyond specific findings, this study also guides further optimization of high-throughput CRISPR experiments in plants.


Genetically modified mouse models used for studying the role of the AT2 receptor in cardiac hypertrophy and heart failure.
The actions of Angiotensin II have been implicated in many cardiovascular conditions. It is widely accepted that the cardiovascular effects of Angiotensin II are mediated by different subtypes of receptors: AT(1) and AT(2). These membrane-bound receptors share a part of their nucleic acid but seem to have different distribution and pathophysiological actions. AT(1) mediates most of the Angiotensin II actions since it is ubiquitously expressed in the cardiovascular system of the normal adult. Moreover AT(2) is highly expressed in the developing fetus but its expression in the cardiovascular system is low and declines after birth. However the expression of AT(2) appears to be modulated by pathological states such as hypertension, myocardial infarction or any pathology associated to tissue remodeling or inflammation. The specific role of this receptor is still unclear and different studies involving in vivo and in vitro experiments have shown conflicting data. It is essential to clarify the role of the AT(2) receptor in the different pathological states as it is a potential site for an effective therapeutic regimen that targets the Angiotensin II system. We will review the different genetically modified mouse models used to study the AT(2) receptor and its association with cardiac hypertrophy and heart failure.


Pannexin-1 in the CNS: Emerging concepts in health and disease.
Pannexin-1 (Panx1) is a large pore membrane channel with unique conduction properties ranging from non-selective ion permeability to the extracellular release of signalling molecules. The release of ATP by Panx1 has been particularly well-characterized with implications in purine signalling across a variety of biological contexts. Panx1 activity is also important in inflammasome formation and the secretion of pro-inflammatory molecules such as interleukin-1β. Within the central nervous system (CNS), Panx1 is expressed on both neurons and glia, and is thought to mediate crosstalk between these cells. A growing body of literature now supports the pathological activity of Panx1 in contributing to disease processes including seizure, stroke, migraine headache and chronic pain. Emerging evidence also reveals a physiological function of Panx1 in regulating neural stem cell survival, neuronal maturation and synaptic plasticity, with possible relevance to normal cognitive functioning. The aim of this review is to summarize the current evidence regarding the roles of Panx1 in the CNS, with emphasis on how putative signalling properties and activation mechanisms of this channel contribute to various physiological and pathophysiological processes.


Identifying metabolic syndrome in African American children using fasting HOMA-IR in place of glucose.
Metabolic syndrome (MetS) is increasing among young people. We compared the use of homeostasis model assessment of insulin resistance (HOMA-IR) with the use of fasting blood glucose to identify MetS in African American children. We performed a cross-sectional analysis of data from a sample of 105 children (45 boys, 60 girls) aged 9 to 13 years with body mass indexes at or above the 85th percentile for age and sex. Waist circumference, blood pressure, and fasting levels of blood glucose, insulin, triglycerides, and high-density lipoprotein cholesterol were measured. We found that HOMA-IR is a stronger indicator of MetS in children than blood glucose. Using HOMA-IR as 1 of the 5 components, we found a 38% prevalence of MetS in this sample of African American children and the proportion of false negatives decreased from 94% with blood glucose alone to 13% with HOMA-IR. The prevalence of MetS was higher in obese than overweight children and higher among girls than boys. Using HOMA-IR was preferred to fasting blood glucose because insulin resistance was more significantly interrelated with the other 4 MetS components.


Subnanosecond mJ eye-safe laser with an intracavity optical parametric oscillator in a shared resonator.
We theoretically verify that the threshold of an intracavity optical parametric oscillator pumped by a passively Q-switched laser is entirely controlled by the bleach of the saturable absorber not by the signal output reflectivity. We use a series of different output couplers to optimize the output performance. With a signal output reflectivity of 15%, we experimentally achieve an efficient subnanosecond eye-safe laser with 3.3 mJ pulse energy and 1.5 MW peak power.


Bell's palsy and herpes simplex virus.
The possible association of some viral infections with the onset of Bell's palsy was examined in a study of 142 patients. The number of probable recent viral infections, as judged by increase in antibody titers or presence of IgM antibodies, did not differ statistically from that found in a sex- and age-matched control group. However, a higher prevalence of herpes simplex virus (HSV) antibodies was found in the patient group with both a complement-fixation (CF) test and a radioimmunoassay (RIA). Moreover, titers of HSV CF antibodies and antibodies against HSV envelope antigens (RIA) were higher in the patient group. The possibility of reactivated HSV infection and transient demyelination of the facial nerve being causatively associated with Bell's palsy is discussed.


Effect of regular exercise on 24-hour arterial pressure in older hypertensive humans.
The experimental goals were to determine if regular low-intensity aerobic exercise reduces 24-hour arterial blood pressure in middle-aged and older (aged 50 years or older) humans with mild diastolic (90-105 mm Hg) essential hypertension and, if so, whether this is accurately reflected by changes in casual recordings made at rest. Fourteen subjects walked 3-4 days/wk for 6 months, with 10 exercising an additional 6 months; 12 other subjects served as nonexercising controls. In the exercising subjects, maximal oxygen consumption increased 7-14% (p less than 0.05) with little or no change in body weight or fat. Conventional casual readings of systolic, mean, and diastolic arterial pressure at rest were lower (5-10 mm Hg, p less than 0.05) in all body positions after 6 months of exercise and changed little thereafter. Casual recordings made during additional circulatory measurements showed 6-month decreases of only half this magnitude and were specific to a particular blood pressure phase and body position; however, all changes were significant after 12 months of exercise. The reductions in arterial pressure at rest were associated with decreases in heart rate (p less than 0.05) and cardiac output (p less than 0.05). Ambulatory-determined 24-hour arterial pressure was unchanged after 6 months of exercise, but mean levels were slightly lower (4 mm Hg, p less than 0.05) after 12 months due to reductions in daytime (7 mm Hg, p less than 0.05) and nighttime (4 mm Hg, NS) systolic pressure; diastolic pressure was unchanged throughout the year of training. In the controls, conventionally recorded casual blood pressure levels were lower after 6 months (p less than 0.05), but no other changes were observed in any other variable over the 12 months of study. We conclude 1) regular low-intensity aerobic exercise at best produces only small reductions in 24-hour levels of arterial pressure in middle-aged and older humans with mild (diastolic) essential hypertension and 2) training-associated changes in casually determined blood pressure at rest are dependent on the measurement conditions and, most importantly, do not necessarily reflect the magnitude or even the direction of changes in arterial pressure throughout an entire day.


The transition of alcohol control in China 1990-2019: Impacts and recommendations.
The harmful use of alcohol is a severe public health issue globally. Chinese per-capita alcohol consumption has increased sharply in recent decades, which has contributed to a rise in alcohol-related problems. In this article we present an analysis of Chinese alcohol policy, beginning with a characterization of alcohol consumption in China followed by an examination of how the nation's alcohol control policy has evolved over the past 30 years, identifying shortcomings and obstacles to improvement. Finally, we present several recommendations informed by the Global Strategy to Reduce the Harmful Use of Alcohol and the SAFER Technical Package-Five Areas of Intervention at National and Subnational Levels (SAFER initiative), to the areas of taxation, alcohol availability, alcohol marketing regulation, and treatment.


Manipulation of the van der Waals Magnet Cr2Ge2Te6 by Spin-Orbit Torques.
We report measurements of current-induced thermoelectric and spin-orbit torque effects within devices in which multilayers of the semiconducting two-dimensional van der Waals magnet Cr2Ge2Te6 (CGT) are integrated with Pt and Ta metal overlayers. We show that the magnetic orientation of the CGT can be detected accurately either electrically (using an anomalous Hall effect) or optically (using magnetic circular dichroism) with good consistency. The samples exhibit large thermoelectric effects, but nevertheless, the spin-orbit torque can be measured quantitatively using the angle-dependent second harmonic Hall technique. For CGT/Pt, we measure the spin-orbit torque efficiency to be similar to conventional metallic-ferromagnet/Pt devices with the same Pt resistivity. The interfacial transparency for spin currents is therefore similar in both classes of devices. Our results demonstrate the promise of incorporating semiconducting 2D magnets within spin-orbitronic and magneto-thermal devices.


Definitions and End Points for Interventional Studies for Arteriovenous Dialysis Access.
This paper is part of the Clinical Trial Endpoints for Dialysis Vascular Access Project of the American Society of Nephrology Kidney Health Initiative. The purpose of this project is to promote research in vascular access by clarifying trial end points which would be best suited to inform decisions in those situations in which supportive clinical data are required. The focus of a portion of the project is directed toward arteriovenous access. There is a potential for interventional studies to be directed toward any of the events that may be associated with an arteriovenous access' evolution throughout its life cycle, which has been divided into five distinct phases. Each one of these has the potential for relatively unique problems. The first three of these correspond to three distinct stages of arteriovenous access development, each one of which has been characterized by objective direct and/or indirect criteria. These are characterized as: stage 1-patent arteriovenous access, stage 2-physiologically mature arteriovenous access, and stage 3-clinically functional arteriovenous access. Once the requirements of a stage 3-clinically functional arteriovenous access have been met, the fourth phase of its life cycle begins. This is the phase of sustained clinical use from which the arteriovenous access may move back and forth between it and the fifth phase, dysfunction. From this phase of its life cycle, the arteriovenous access requires a maintenance procedure to preserve or restore sustained clinical use. Using these definitions, clinical trial end points appropriate to the various phases that characterize the evolution of the arteriovenous access life cycle have been identified. It is anticipated that by using these definitions and potential end points, clinical trials can be designed that more closely correlate with the goals of the intervention and provide appropriate supportive data for clinical, regulatory, and coverage decisions.


Cooperation between prokaryotic (Lde) and eukaryotic (MRP) efflux transporters in J774 macrophages infected with Listeria monocytogenes: studies with ciprofloxacin and moxifloxacin.
Antibiotic efflux is observed in both eukaryotic and prokaryotic cells, modulating accumulation and resistance. The present study examines whether eukaryotic and prokaryotic fluoroquinolone transporters can cooperate in the context of an intracellular infection. We have used (i) J774 macrophages (comparing a ciprofloxacin-resistant cell line overexpressing an MRP-like transporter with wild-type cells with basal expression), (ii) Listeria monocytogenes (comparing a clinical isolate [CLIP21369] displaying ciprofloxacin resistance associated with overexpression of the Lde efflux system with a wild-type strain [EGD]), (iii) ciprofloxacin (substrate of both Lde and MRP) and moxifloxacin (nonsubstrate), and (iv) probenecid and reserpine (preferential inhibitors of MRP and Lde, respectively). The ciprofloxacin MICs for EGD were unaffected by reserpine, while those for CLIP21369 were decreased approximately fourfold (and made similar to those of EGD). Neither probenecid nor reserpine affected the moxifloxacin MICs against EGD or CLIP21369. In dose-response studies (0.01x to 100x MIC) in broth, reserpine fully restored the susceptibility of CLIP21369 to ciprofloxacin (no effect on EGD) but did not influence the activity of moxifloxacin. In studies with intracellular bacteria, reserpine, probenecid, and their combination increased the activity of ciprofloxacin in wild-type and ciprofloxacin-resistant macrophages in parallel with an increase in ciprofloxacin accumulation in macrophages for EGD and an increase in accumulation and decrease in MIC (in broth) for CLIP21369. Moxifloxacin accumulation and intracellular activity were consistently not affected by the inhibitors. A bacterial efflux pump may thus actively cooperate with a eukaryotic efflux transporter to reduce the activity of a common substrate (ciprofloxacin) toward an intracellular bacterial target.


Dimethyl sulfoxide prevents 7β-hydroxycholesterol-induced apoptosis by preserving lysosomes and mitochondria.
Dimethyl sulfoxide (DMSO) is a widely used vehicle for water-insoluble substances and exerts a wide range of pharmacologic effects including anti-inflammatory and free radical scavenging properties. Additionally, in an animal model, DMSO inhibited cholesterol-induced atherosclerosis. Despite such profound pharmacologic effects, mechanisms at the cellular level are not well understood. Atherogenic oxysterols, especially 7-oxysterols, are potent inducers of oxidative stress, cell apoptosis, and are elevated in human atherosclerotic lesions. In this study, we first investigated the effect of DMSO on 7beta-hydroxycholesterol-induced apoptosis of U937 cells and then focused on its influences on production of reactive oxygen species, lysosomal, and mitochondrial membrane permeability. Our results revealed that DMSO protected U937 cells against 7beta-hydroxycholesterol-induced cell death by preventing lysosomal and mitochondrial membrane permeabilization and reactive oxygen species production. Our results also emphasize the necessity for appropriate solvent control groups in experimental models in which DMSO has been used to examine drug effect or identify pathways.


Coronary and carotid atherosclerosis: how useful is the imaging?
The recent advancement of imaging modalities has made possible visualization of atherosclerosis disease in all phases of its development. Markers of subclinical atherosclerosis or even the most advanced plaque features are acquired by invasive (IVUS, OCT) and non-invasive imaging modalities (US, MRI, CTA). Determining plaques prone to rupture (vulnerable plaques) might help to identify patients at risk for myocardial infarction or stroke. The most accepted features of plaque vulnerability include: thin cap fibroatheroma, large lipid core, intimal spotty calcification, positive remodeling and intraplaque neovascularizations. Today, research is focusing on finding imaging techniques that are less invasive, less radiation and can detect most of the vulnerable plaque features. While, carotid atherosclerosis can be visualized using noninvasive imaging, such as US, MRI and CT, imaging plaque feature in coronary arteries needs invasive imaging modalities. However, atherosclerosis is a systemic disease with plaque development simultaneously in different arteries and data acquisition in carotid arteries can add useful information for prediction of coronary events.


The single chain limit of structural relaxation in a polyolefin blend.
The influence of composition on component dynamics and relevant static properties in a miscible polymer blend is investigated using molecular dynamics simulation. Emphasis is placed on dynamics in the single chain dilution limit, as this limit isolates the role of inherent component mobility in the polymer's dynamic behavior when placed in a blend. For our systems, a biased local concentration affecting dynamics must arise primarily from chain connectivity, which is quantified by the self-concentration, because concentration fluctuations are minimized due to restraints on chain lengths arising from simulation considerations. The polyolefins simulated [poly(ethylene-propylene) (PEP) and poly(ethylene-butene) (PEB)] have similar structures and glass transition temperatures, and all interactions are dispersive in nature. We find that the dependence of dynamics upon composition differs between the two materials. Specifically, PEB (slower component) is more influenced by the environment than PEP. This is linked to a smaller self-concentration for PEB than PEP. We examine the accuracy of the Lodge-McLeish model (which is based on chain connectivity acting over the Kuhn segment length) in predicting simulation results for effective concentration. The model predicts the simulation results with high accuracy when the model's single parameter, the self-concentration, is calculated from simulation data. However, when utilizing the theoretical prediction of the self-concentration the model is not quantitatively accurate. The ability of the model to link the simulated self-concentration with biased local compositions at the Kuhn segment length provides strong support for the claim that chain connectivity is the leading cause of distinct mobility in polymer blends. Additionally, the direct link between the willingness of a polymer to be influenced by the environment and the value of the self-concentration emphasizes the importance of the chain connectivity. Furthermore, these findings are evidence that the Kuhn segment length is the relevant length scale controlling segmental dynamics.


Nerve injuries in the hand.
Nerve injuries of the hand are an important problem for the emergency physician because most are diagnosed in the emergency setting. Successful diagnosis and treatment are critical if the injured patient is to regain full hand function. Emergency department management relies on the knowledge of hand and forearm functional anatomy and techniques for diagnosing loss of nerve function. Once a nerve injury has been diagnosed, issues such as primary versus secondary repair must be discussed with the hand surgeon. The hand surgeon must consider whether to pursue an epineurial or fascicular repair and must use additional techniques for diagnosing return of nervous function. Lack of nerve regeneration, chronic paresthesias, neuromas, and sympathetic dystrophy can complicate the successful return of hand nerve function after injury.


Complementary clinical effects of topical tightening treatment in conjunction with a radiofrequency procedure.
Abstract Background: Skin laxity and cellulite on the buttocks and thighs are two common cosmetic concerns. Skin tightening with radiofrequency (RF) devices has become increasingly popular. The purpose of this study is to evaluate the efficacy and safety of a topical skin laxity tightening agent when used in combination with an RF device. A double-blinded, randomized clinical trial enrolled twenty females with mild-to-moderate skin laxity on the posterior thighs/buttocks. Each subject underwent two monthly treatments with an RF source (Alma Accent) to both legs. Subjects were then randomized to apply a topical agent (Skinceuticals Body Tightening Concentrate) twice daily to only one designated thigh/buttock throughout the eight-week duration of the study. All subjects were evaluated for improvement in lifting, skin tone, radiance, firmness/tightness, skin texture, and overall appearance based on photographic evaluation by blinded investigators at 12 weeks following the final RF treatment. A statistically significant improvement was found in the overall appearance on both sides treated with the RF device when compared to baseline. However, the area treated with the topical agent showed a statistically significantly greater degree of improvement than the side where no topical agent was applied. No adverse effects were reported. The use of a novel skin tightening agent used after RF procedures is both safe and effective for treatment of skin laxity on the buttocks and thighs. Combined therapy leads to a better result.


[Education for self-administered antibiotic therapy: a pragmatic and ethical alternative for the treatment of STDs for the street youth of Kinshasa in the Democratic Republic of the Congo (RDC)].
All healthcare providers decide in someone else's place, for someone else. In doing so, they take their place in a long long tradition, that of medical paternalism. Patients are treated as children, incapable of making decisions about themselves. How then are we supposed to deal with patients like the street children of the Democratic Republic of the Congo, who are not part of our health-care system, who refuse care and prescriptions? Their refusal of caregivers forces us to seek strategies to dispel the conflicts, adapt outselves to the situation (self-medication, drug sales outside of dispensaries, etc.), but especially to rethink the relation between caregivers and patients. This does not mean abandoning the authoritarian patriarchal model for total relativism; the use of drugs such as antibiotics is and must remain surrounded by all the precautions necessary to avoid the further development of resistance; it does mean training and informing. The task facing us is that of health education and promotion, a long and continuous process, centered on patients and integrated with their care, aimed at making them capable of managing their disease. This procedure is part of a pragmatic approach: beyond the asymmetry involved in any relationship of power, it is essential to establish informed confidence, to look for adhesion and not constraint. Only this pragmatism can incite young people with sexually transmitted diseases (STDs) to use modern medicine and comply with the dosage instructions. Effective treatment of STDs is, according to WHO, one of the most powerful weapons in the battle against AIDS transmission.


Incidental Lewy body disease: clinical comparison to a control cohort.
Limited clinical information has been published on cases pathologically diagnosed with incidental Lewy body disease (ILBD). Standardized, longitudinal movement and cognitive data was collected on a cohort of subjects enrolled in the Sun Health Research Institute Brain and Body Donation Program. Of 277 autopsied subjects who had antemortem clinical evaluations within the previous 3 years, 76 did not have Parkinson's disease, a related disorder, or dementia of which 15 (20%) had ILBD. Minor extrapyramidal signs were common in subjects with and without ILBD. Cognitive testing revealed an abnormality in the ILBD group in the Trails B test only. ILBD cases had olfactory dysfunction; however, sample size was very small. This preliminary report revealed ILBD cases have movement and cognitive findings that for the most part were not out of proportion to similarly assessed and age-similar cases without Lewy bodies. Larger sample size is needed to have the power to better assess group differences.


Identifying potential receptors and routes of contaminant exposure in the traditional territory of the Ouje-Bougoumou Cree: land use and a geographical information system.
Great concern has been raised with respect to the 13 traplines that constitute the traditional territory of the Ouje-Bougoumou Cree located in the James Bay region of northern Quebec, Canada, with respect to mine wastes originating from three local mines. As a result, an "Integrative Risk Assessment" was initiated consisting of three interrelated components: a comprehensive human health study, an assessment of the existing ecological/environmental database, and a land use/potential sites of concern study. In this paper, we document past and present land use in the traditional territory of the Ouje-Bougoumou Cree for 72 heads of households, including 13 tallymen, and use a Geographic Information System (GIS) to layer harvest/hunting and gathering/collecting data over known mining areas and potential sites of concern. In this way, potential receptors of contamination and routes of human exposure were identified. Areas of overlap with respect to land use activity and mining operations were relatively extensive for certain harvesting activities (e.g., beaver, Castor canadensis and various species of game birds), less so for fish harvesting (all species) and water collection, and relatively restrictive for large mammal harvesting and collection of fire wood (and other collection activities). Potential receptors of contaminants associated with mining activity (e.g., fish and small mammals) and potential routes of exposure (e.g., ingestion of contaminated game and drinking of contaminated water) were identified.


Whole genome detection of sequence and structural polymorphism in six diverse horses.
The domesticated horse has played a unique role in human history, serving not just as a source of animal protein, but also as a catalyst for long-distance migration and military conquest. As a result, the horse developed unique physiological adaptations to meet the demands of both their climatic environment and their relationship with man. Completed in 2009, the first domesticated horse reference genome assembly (EquCab 2.0) produced most of the publicly available genetic variations annotations in this species. Yet, there are around 400 geographically and physiologically diverse breeds of horse. To enrich the current collection of genetic variants in the horse, we sequenced whole genomes from six horses of six different breeds: an American Miniature, a Percheron, an Arabian, a Mangalarga Marchador, a Native Mongolian Chakouyi, and a Tennessee Walking Horse, and mapped them to EquCab3.0 genome. Aside from extreme contrasts in body size, these breeds originate from diverse global locations and each possess unique adaptive physiology. A total of 1.3 billion reads were generated for the six horses with coverage between 15x to 24x per horse. After applying rigorous filtration, we identified and functionally annotated 17,514,723 Single Nucleotide Polymorphisms (SNPs), and 1,923,693 Insertions/Deletions (INDELs), as well as an average of 1,540 Copy Number Variations (CNVs) and 3,321 Structural Variations (SVs) per horse. Our results revealed putative functional variants including genes associated with size variation like LCORL gene (found in all horses), ZFAT in the Arabian, American Miniature and Percheron horses and ANKRD1 in the Native Mongolian Chakouyi horse. We detected a copy number variation in the Latherin gene that may be the result of evolutionary selection impacting thermoregulation by sweating, an important component of athleticism and heat tolerance. The newly discovered variants were formatted into user-friendly browser tracks and will provide a foundational database for future studies of the genetic underpinnings of diverse phenotypes within the horse.


Recruitment via social media: advantages and potential biases.
Adolescent and young adult (AYA) cancer survivors are under-represented in research. Social media is increasingly used for recruitment given its ability to reach large audiences. Differences in participant characteristics and potential biases due to recruitment source are not well understood. This study aimed to: (a) compare recruitment strategies (hospital-based v. social media) in enrollment metrics, and (b) among enrolled participants, evaluate group differences in patient characteristics and patient reported outcomes (PROs). Preliminary data from a cancer and fertility study with female AYAs were evaluated. Hospital-based recruitment used electronic medical records (EMR) to identify eligible patients. Social media recruitment involved posting on partner organizations' social media outlets. PROs included validated measures related to the parent study. Descriptive statistics evaluated recruitment metrics. Independent samples t-tests and chi-square identified differences in participant characteristics and PROs based on recruitment. Social media yielded a higher enrollment rate (37%; n = 54/146) compared with hospital-based recruitment (7%; n = 21/289) and required fewer study resources. Compared with hospital-based recruitment, participants from social media were more likely to be White (p = 0.01), with a longer time since treatment (p = 0.03); and reported higher levels of reproductive concern (p = 0.004) and negative mood (p = 0.02), and more negative illness perceptions (ps < 0.05). Recruitment via social media may be a more effective and efficient strategy compared with hospital-based methods. However, group differences were identified that could bias findings and limit generalizability. Advantages of social media should be considered with an understanding of how methodology may impact enrollment and results.


Regulation of the dual specificity protein phosphatase, DsPTP1, through interactions with calmodulin.
Reversible phosphorylation is a key mechanism for the control of intercellular events in eukaryotic cells. In animal cells, Ca2+/CaM-dependent protein phosphorylation and dephosphorylation are implicated in the regulation of a number of cellular processes. However, little is known on the functions of Ca2+/CaM-dependent protein kinases and phosphatases in Ca2+ signaling in plants. From an Arabidopsis expression library, we isolated cDNA encoding a dual specificity protein phosphatase 1, which is capable of hydrolyzing both phosphoserine/threonine and phosphotyrosine residues of the substrates. Using a gel overlay assay, we identified two Ca2+-dependent CaM binding domains (CaMBDI in the N terminus and CaMBDII in the C terminus). Specific binding of CaM to two CaMBD was confirmed by site-directed mutagenesis, a gel mobility shift assay, and a competition assay using a Ca2+/CaM-dependent enzyme. At increasing concentrations of CaM, the biochemical activity of dual specificity protein phosphatase 1 on the p-nitrophenyl phosphate (pNPP) substrate was increased, whereas activity on the phosphotyrosine of myelin basic protein (MBP) was inhibited. Our results collectively indicate that calmodulin differentially regulates the activity of protein phosphatase, dependent on the substrate. Based on these findings, we propose that the Ca2+ signaling pathway is mediated by CaM cross-talks with a protein phosphorylation signal pathway in plants via protein dephosphorylation.


Hyaluronate capsule prevents attachment of group A streptococci to mouse peritoneal macrophages.
The antiphagocytic properties of the streptococcal hyaluronic acid capsule were explored in a system in which binding of the organism to the phagocyte, but not ingestion, could occur. The capsule was found to be highly effective in preventing attachment of two strains of group A streptococci to mouse peritoneal macrophages. Variation in attachment with phase of growth in broth culture (low in early log phase, high in late-log phase and thereafter) could be accounted for by production and subsequent loss of capsule. Hyaluronidase treatment removed the capsule and increased adherence; treatment with proteolytic enzymes removed M protein and decreased resistance to phagocytosis in whole human blood but did not remove the capsule or increase adherence to the mouse peritoneal macrophages.


Nuclear DDX3 expression predicts poor outcome in colorectal and breast cancer.
DEAD box protein 3 (DDX3) is an RNA helicase with oncogenic properties that shuttles between the cytoplasm and nucleus. The majority of DDX3 is found in the cytoplasm, but a subset of tumors has distinct nuclear DDX3 localization of yet unknown biological significance. This study aimed to evaluate the significance of and mechanisms behind nuclear DDX3 expression in colorectal and breast cancer. Expression of nuclear DDX3 and the nuclear exporter chromosome region maintenance 1 (CRM1) was evaluated by immunohistochemistry in 304 colorectal and 292 breast cancer patient samples. Correlations between the subcellular localization of DDX3 and CRM1 and the difference in overall survival between patients with and without nuclear DDX3 were studied. In addition, DDX3 mutants were created for in vitro evaluation of the mechanism behind nuclear retention of DDX3. DDX3 was present in the nucleus of 35% of colorectal and 48% of breast cancer patient samples and was particularly strong in the nucleolus. Nuclear DDX3 correlated with worse overall survival in both colorectal (hazard ratio [HR] 2.34, P<0.001) and breast cancer (HR 2.39, P=0.004) patients. Colorectal cancers with nuclear DDX3 expression more often had cytoplasmic expression of the nuclear exporter CRM1 (relative risk 1.67, P=0.04). In vitro analysis of DDX3 deletion mutants demonstrated that CRM1-mediated export was most dependent on the N-terminal nuclear export signal. Overall, we conclude that nuclear DDX3 is partially CRM1-mediated and predicts worse survival in colorectal and breast cancer patients, putting it forward as a target for therapeutic intervention with DDX3 inhibitors under development in these cancer types.


Palatal surgery in a transoral robotic setting (TORS): preliminary results of a retrospective comparison between uvulopalatopharyngoplasty (UPPP), expansion sphincter pharyngoplasty (ESP) and barbed repositioning pharyngoplasty (BRP).
It has become increasingly clear in the past decade that surgical management of obstructive sleep apnoea hypopnoea syndrome (OSAHS) is most successfully managed with multilevel surgery. We evaluated the outcomes of multilevel interventions comparing three different palatal techniques added to TORS: uvulopalatopharyngoplasty (UPPP), a modified expansion sphincter pharyngoplasty (ESP), inspired by the Pang expansion sphincter pharyngoplasty technique and the latest barbed repositioning pharyngoplasty (BRP). Thirty patients were retrospectively evaluated. Ten patients underwent UPPP by Fairbanks, 10 BRP and 10 a modified ESP already described. All patients underwent TORS, tonsillectomy and septo-turbinoplasty. For all cases, the following data were retrieved and revaluated: preoperative and postoperative apnoea-hypopnoea index (AHI), preoperative and postoperative Epworth Sleepiness Scale (ESS), pain visual analogue scale (VAS; 0-10) for the first 5 days postoperatively, palatal operative time for each surgical technique, discharge date and complication types and rate. Both BRP and ESP resulted in better postoperative AHI values and higher surgical success rates in comparison with UPPP. On the other hand, BRP was not more effective than ESP. ESP surgery time was significantly higher than UPPP, while BRP was the quickest procedure. In summary, ESP and BRP seem to be more effective than UPPP in a multilevel surgical robotic setting. However, being quicker, easy to learn and with a low rate of complications, BRP is a safe, effective and promising option for treatment of OSAHS patients.


[Clinical and experimental study of kidney tonics plus whole embryo extract in treatment of renal anemia].
According to the therapeutic principles, reinforcing Kidney-Qi and nourishing the blood were used in treating renal anemia, Renal Anemia Granule No. I and No. II (RAG I and II) were applied in treating patients of renal anemia, the former was a Yin reinforcing agent and the latter a Yang tonics principally. Thirty patients were treated according to the Syndrome Differentiation of TCM and divided into two groups: (1) The combined group, oral liquid of the whole embryo extract was given additionally. (2) The discombined group, no additional drug was given. Patients after treatment all showed significant improvements in serum erythropoietin and hemoglobin (P < 0.05). Experimental study in animal models also displayed the hemoglobin of treated group increased more than that of control group (P < 0.05), which might be related to the enhancement of hematopoietic function of bone marrow.


Extracellular Vesicles for the Diagnosis and Treatment of Parkinson's Disease.
Extracellular vesicles (EVs) shed by neurons and glia in the central nervous system carry a cargo of specific bioactive molecules, facilitating intercellular communication. However, in neurodegenerative disease conditions, EVs carry pathological miRNAs and/or proteins involved in spreading the disease. Such EVs are also found in the cerebrospinal fluid (CSF) or the circulating blood, the characterization of which could identify biomarkers linked to specific neurodegenerative diseases. Moreover, EVs secreted by various stem/progenitor cells carry therapeutic miRNAs and proteins, which have shown promise to alleviate symptoms and slow down the progression of neurodegenerative diseases. The ability of exogenously administered EVs to easily cross the blood-brain barrier with no risk for thrombosis and incorporate into neurons and glia has also opened up the possibility of using nano-sized EVs as carriers of therapeutic drugs or bioactive proteins. This review summarizes the role and function of EVs in alpha-synuclein-mediated neurodegeneration and the spread of alpha-synuclein from neurons to glia, leading to the activation of the inflammatory response in Parkinson's disease (PD). Moreover, the promise of brain-derived EVs in the CSF and the circulating blood for biomarker discovery and the efficacy of stem/progenitor cell-derived EVs or EVs loaded with bioactive molecules such as dopamine, catalase, curcumin, and siRNAs, in alleviating Parkinsonian symptoms are discussed.


Identification of chemoradiation-resistant osteosarcoma stem cells using an imaging system for proteasome activity.
Osteosarcoma is the most common primary bone malignancy in pediatric and adolescent populations. Recurrence and metastatic potential can be due to a subpopulation of cells with stem cell-like characteristics, such as tumor-initiating cells (TICs), which maintain the capacity to regenerate entire tumors. Targeting the TICs in osteosarcoma is a promising avenue for the development of new therapies for this devastating disease. TICs are usually quiescent with a low protein turnover, decreased metabolism, and downregulation of proteasome activity. Recently, cancer cells with low proteasome activity have been identified as TICs in several types of cancer. We stably infected two osteosarcoma cell lines, MG-63 and U2-OS, with an expression vector for a fusion protein between the green fluorescent protein, ZsGreen, and the C-terminal degron of the murine ornithine decarboxylase to monitor the 26S proteasome activity in living cells. We separated the osteosarcoma cells with low proteasome activity using fluorescence-activated cell sorting (FACS) and verified whether these ZsGreen+ cells had TIC-like properties. The ZsGreen+ cells showed enhanced sphere formation capacity and underwent asymmetric divisions into ZsGreen+ and ZsGreen- cells, whereas ZsGreen- cells underwent only symmetric divisions into ZsGreen- cells. Moreover, the ZsGreen+ cells were more chemo- and radioresistant. Thus, the present study demonstrated that chemoradiation-resistant TICs can be visualized by this system and suggested the rationale for further study of osteosarcoma stem cells.


Antihypertensives and Statin Therapy for Primary Stroke Prevention: A Secondary Analysis of the HOPE-3 Trial.
The HOPE-3 trial (Heart Outcomes Prevention Evaluation–3) found that antihypertensive therapy combined with a statin reduced first stroke among people at intermediate cardiovascular risk. We report secondary analyses of stroke outcomes by stroke subtype, predictors, treatment effects in key subgroups. Using a 2-by-2 factorial design, 12 705 participants from 21 countries with vascular risk factors but without overt cardiovascular disease were randomized to candesartan 16 mg plus hydrochlorothiazide 12.5 mg daily or placebo and to rosuvastatin 10 mg daily or placebo. The effect of the interventions on stroke subtypes was assessed. Participants were 66 years old and 46% were women. Baseline blood pressure (138/82 mm Hg) was reduced by 6.0/3.0 mm Hg and LDL-C (low-density lipoprotein cholesterol; 3.3 mmol/L) was reduced by 0.90 mmol/L on active treatment. During 5.6 years of follow-up, 169 strokes occurred (117 ischemic, 29 hemorrhagic, 23 undetermined). Blood pressure lowering did not significantly reduce stroke (hazard ratio [HR], 0.80 [95% CI, 0.59–1.08]), ischemic stroke (HR, 0.80 [95% CI, 0.55–1.15]), hemorrhagic stroke (HR, 0.71 [95% CI, 0.34–1.48]), or strokes of undetermined origin (HR, 0.92 [95% CI, 0.41–2.08]). Rosuvastatin significantly reduced strokes (HR, 0.70 [95% CI, 0.52–0.95]), with reductions mainly in ischemic stroke (HR, 0.53 [95% CI, 0.37–0.78]) but did not significantly affect hemorrhagic (HR, 1.22 [95% CI, 0.59–2.54]) or strokes of undetermined origin (HR, 1.29 [95% CI, 0.57–2.95]). The combination of both interventions compared with double placebo substantially and significantly reduced strokes (HR, 0.56 [95% CI, 0.36–0.87]) and ischemic strokes (HR, 0.41 [95% CI, 0.23–0.72]). Among people at intermediate cardiovascular risk but without overt cardiovascular disease, rosuvastatin 10 mg daily significantly reduced first stroke. Blood pressure lowering combined with rosuvastatin reduced ischemic stroke by 59%. Both therapies are safe and generally well tolerated. URL: https://www.clinicaltrials.gov; Unique identifier: NCT00468923.


Differences in eye movements for face recognition between Canadian and Chinese participants are not modulated by social orientation.
Face recognition strategies do not generalize across individuals. Many studies have reported robust cultural differences between West Europeans/North Americans and East Asians in eye movement strategies during face recognition. The social orientation hypothesis posits that individualistic vs. collectivistic (IND/COL) value systems, respectively defining West European/North American and East Asian societies, would be at the root of many cultural differences in visual perception. Whether social orientation is also responsible for such cultural contrast in face recognition remains to be clarified. To this aim, we conducted two experiments with West European/North American and Chinese observers. In Experiment 1, we probed the existence of a link between IND/COL social values and eye movements during face recognition, by using an IND/COL priming paradigm. In Experiment 2, we dissected the latter relationship in greater depth, by using two IND/COL questionnaires, including subdimensions to those concepts. In both studies, cultural differences in fixation patterns were revealed between West European/North American and East Asian observers. Priming IND/COL values did not modulate eye movement visual sampling strategies, and only specific subdimensions of the IND/COL questionnaires were associated with distinct eye-movement patterns. Altogether, we show that the typical contrast between IND/COL cannot fully account for cultural differences in eye movement strategies for face recognition. Cultural differences in eye movements for faces might originate from mechanisms distinct from social orientation.


Tyrosine Kinase Inhibitor Therapies in Chronic Myeloid Leukemia: Effects on Clinical Characteristics and Triglyceride-to-High Density Lipoprotein Cholesterol Ratio.
Tyrosine kinase inhibitors (TKIs) have dramatically improved chronic myeloid leukemia (CML) prognosis. However, TKIs are associated with dyslipidemia and impaired glucosehomeostasis. Triglyceride-to-high-density lipoprotein cholesterol ratio (TG/HDL-C) is proposed to be an indicator of insulin resistance and atherogenic index, but there is no research on TG/HDL-C alterations in patients receiving TKIs for CML. We aimed to evaluate relationships between TKI type/count, clinical characteristics, and laboratory results (particularly TG/HDL-C) in CML patients. A total of 104 patients with chronic phase CML were enrolled in the study. All patients received initial imatinib therapy at 400 mg daily, the type or dose of TKI was then changed according to treatment response and clinical outcomes. Patients were compared with respect to TG/HDL-C categorization (>2.5 versus <2.5), number of TKIs used, and use of imatinib as the only TKI. The median TG/HDL-C was 2.82 (1.03-17.33) and this ratio was higher than 2.5 in 59 (56.7%) patients. Patients with high TG/HDL-C had a significantly higher age than patients with low values (P < 0.001). Recipients of more than one TKI had higher EUTOS risk score and white blood cell (WBC) count (P < 0.05). Recipients of imatinib as the only TKI had higher age, low EOTUS risk score, low WBC, and low neutrophil count (all, P < 0.05). TG/HDL-C values were not associated with the number of different TKIs used or the use of imatinib only in chronic-phase patients with CML. Further large-scale prospective studies are needed to determine whether TG/HDL-C can be used for diagnostic or prognostic purposes in TKI recipients.


The significance of plagioclase textures in mid-ocean ridge basalt (Gakkel Ridge, Arctic Ocean).
Textures and compositions of minerals can be used to infer the physiochemical conditions present within magmatic systems. Given that plagioclase is an abundant phase in many magmatic systems, understanding the link between texture and process is vital. Here, we present a database of textural and compositional data for > 1800 plagioclase crystals in mid-ocean ridge basalt from the Gakkel Ridge (Arctic Ocean) to investigate the physiochemical conditions and processes that govern the formation of plagioclase textures and compositions. The Gakkel basalts have high modal crystal contents (up to 50%). The crystal cargo is complex, with both individual plagioclase and glomerocrysts showing large variations in crystal habit, zoning and resorption. The most common types of zoning are reverse and patchy; we attribute patchy zoning to infilling following either skeletal growth or resorption. Resorption is abundant, with multiple resorption events commonly present in a single crystal, and results from both magmatic recharge and decompression. Periods of strong undercooling, distinct to quench crystallisation, are indicated by matured skeletal crystals and thin normally zoned melt inclusion-rich bands following resorption. Individual samples often contain diverse textural and compositional plagioclase groups. Furthermore, most plagioclase is not in equilibrium with its host melt. Finally, the porous open structures of some glomerocrysts suggest that they represent pieces of entrained disaggregated mush. We interpret this to indicate that the crystal cargo is not generally phenocrystic in origin. Instead, plagioclase crystals that formed in different parts of a mush-dominated plumbing system were entrained into ascending melts. The textures of individual crystals are a function of their respective histories of (under)cooling, magma mixing and decompression. The morphologies of melt inclusion trapped in the plagioclase crystals are associated with specific host crystal textures, suggesting a link between plagioclase crystallisation processes and melt inclusion entrapment. The database of plagioclase presented herein may serve as a template for the interpretation of plagioclase textures in magmatic systems elsewhere.


Arthroscopic inferior capsular shift: long-term follow-up.
Neer and Foster's open inferior capsular shift to treat acquired cases of anteroinferior shoulder instability due to an overstretched and redundant capsule is described with good results. Recently, new arthroscopic techniques were described to manage this problem. To assess the results of a new arthroscopic reinforced inferior capsular shift technique based on Neer and Foster's open inferior capsular shift. Case series; Level of evidence, 4. This new technique of arthroscopic inferior capsular shift was used to treat 108 patients with anteroinferior shoulder instability due to capsular redundancy as confirmed clinically and during arthroscopy. It reduces the size of the redundant capsular pouch and reinforces the thinned-out capsule. Intraoperatively, patients with associated labral tears (n = 25) and patients with open rotator intervals (n = 8) were excluded, and only 75 patients with pure capsular redundancy were included in this study. Patients were followed for a minimum of 7 years. All 75 patients had patulous and redundant capsules. Three patients (4.0%) had a redislocation after a significant trauma. The range of motion preoperatively was 168.1° ± 7.5° in forward elevation, 64.7° ± 7.9° in external rotation, and T5.0 ± T0.8 in internal rotation. Postoperatively, it was 167.2° ± 5.8° in forward elevation, 59.95° ± 4.9° in external rotation, and T7.1 ± T1.0 in internal rotation. The American Shoulder and Elbow Surgeons (ASES) (70.76 to 97.53; P < .001), Constant (90.02 to 99.24; P < .001), and University of California, Los Angeles (UCLA) (21.97 to 33.84; P < .001) scores demonstrated significant improvement postoperatively. This novel technique of arthroscopic capsular shift addresses the problem of capsular redundancy present in many cases of anteroinferior shoulder instability. It tries to achieve a capsular shift based on the principles of Neer. The long-term results are very good.


Increased synthesis and deposition of extracellular matrix proteins leads to endoplasmic reticulum stress in the trabecular meshwork.
Increased synthesis and deposition of extracellular matrix (ECM) proteins in the trabecular meshwork (TM) is associated with TM dysfunction and intraocular pressure (IOP) elevation in glaucoma. However, it is not understood how ECM accumulation leads to TM dysfunction and IOP elevation. Using a mouse model of glucocorticoid (GC)-induced glaucoma, primary human TM cells and human post-mortem TM tissues, we show that increased ECM accumulation leads to endoplasmic reticulum (ER) stress in the TM. The potent GC, dexamethasone (Dex) increased the secretory protein load of ECM proteins in the ER of TM cells, inducing ER stress. Reduction of fibronectin, a major regulator of ECM structure, prevented ER stress in Dex-treated TM cells. Overexpression of fibronectin via treatment with cellular fibronectin also induced chronic ER stress in primary human TM cells. Primary human TM cells grown on ECM derived from Dex-treated TM cells induced ER stress markers. TM cells were more prone to ER stress from ECM accumulation compared to other ocular cell types. Moreover, increased co-localization of ECM proteins with ER stress markers was observed in human post-mortem glaucomatous TM tissues. These data indicate that ER stress is associated with increased ECM accumulation in mouse and human glaucomatous TM tissues.


Turbulence induces metabolically costly behaviors and inhibits food capture in oyster larvae, causing net energy loss.
Planktotrophic invertebrate larvae require energy to develop, disperse and settle successfully, and it is unknown how their energetics are impacted by turbulence. Ciliated larvae gain metabolic energy from their phytoplankton food to offset the energetic costs of growth, development and ciliary activity for swimming and feeding. Turbulence may affect the energetic balance by inducing behaviors that alter the metabolic costs and efficiency of swimming, by raising the encounter rate with food particles and by inhibiting food capture. We used experiments and an empirical model to quantify the net rate of energy gain, swimming efficiency and food capture efficiency for eyed oyster larvae (Crassostrea virginica) in turbulence. At dissipation rates representative of coastal waters, larvae lost energy even when food concentrations were very high. Both feeding activity and turbulence-induced behaviors incurred high metabolic costs. Swimming efficiency was concave up versus dissipation rate, suggesting that ciliary activity for food handling became more costly while swimming became more efficient with turbulence intensity. Though counter-intuitive, swimming may have become more efficient in turbulence because vorticity-induced rotation caused larvae to swim more horizontally, which requires less effort than swimming vertically against the pull of gravity. Overall, however, larvae failed to offset high activity costs with food energy gains because turbulence reduced food capture efficiency more than it enhanced food encounter rates. Younger, smaller larvae may have some energetic advantages, but competent larvae would lose energy at turbulence intensities they experience frequently, suggesting that turbulence-induced starvation may account for much of oysters' high larval mortality.


Mechanobiological bone reaction quantified by positron emission tomography.
While nuclear medicine has been proven clinically effective for examination of the change in bone turnover as a result of stress injury, quantitative correlation between tracer uptake and mechanical stimulation in the human jawbone remains unclear. This study aimed to investigate the relationship between bone metabolism observed by 18F-fluoride positron emission tomography (PET) images and mechanical stimuli obtained by finite element analysis (FEA) in the residual ridge induced by the insertion of a removable partial denture (RPD). An 18F-fluoride PET/CT (computerized tomography) scan was performed to assess the change of bone metabolism in the residual ridge under the denture before and after RPD treatment. Corresponding patient-specific 3D finite element (FE) models were created from CT images. Boundary conditions were prescribed by the modeling of condylar contacts, and muscular forces were derived from the occlusal forces measured in vivo to generate mechanobiological reactions. Different mechanobiological stimuli, e.g., equivalent von Mises stress (VMS), equivalent strain (EQV), and strain energy density (SED), determined from nonlinear FEA, were quantified and compared with the standardized uptake values (SUVs) of PET. Application of increased occlusal force after RPD insertion induced higher mechanical stimuli in the residual bone. Accordingly, SUV increased in the region of residual ridge with higher mechanical stimuli. Thus, with SUV, a clear correlation was observed with VMS and SED in the cancellous bone, especially after RPD insertion (R(2) > 0.8, P < 0.001). This study revealed a good correlation between bone metabolism and mechanical stimuli induced by RPD insertion. From this patient-specific study, it was shown that metabolic change detected by PET in the loaded bone, in a much shorter duration than conventional x-ray assessment, is associated with mechanical stimuli. The nondestructive nature of PET/CT scans and FEA could potentially provide a new method for clinical examination and monitoring of prosthetically driven bone remodeling.


Immunological Investigation for the Presence of Lunasin, a Chemopreventive Soybean Peptide, in the Seeds of Diverse Plants.
Lunasin, a 44 amino acid soybean bioactive peptide, exhibits anticancer and anti-inflammatory properties. All soybean varieties that have been examined contain lunasin. It has also been reported in a few other plant species including amaranth, black nightshade, wheat, barley, rye, and triticale. Interestingly, detailed searches of transcriptome and DNA sequence databases of cereals failed to identify lunasin-coding sequences, raising questions about the authenticity of lunasin in cereals. To clarify the presence or absence of lunasin in cereals and other plant species, an immunological investigation was conducted utilizing polyclonal antibodies raised against the first 20 amino acid N-terminal peptide (SKWQHQQDSCRKQLQGVNLT) and a 15 amino acid C-terminal peptide (CEKHIMEKIQGRGDD) of lunasin. Protein blot analyses revealed the presence of proteins from several plants that reacted against the lunasin N-terminal peptide antibodies. However, the same proteins failed to react against the lunasin C-terminal peptide antibodies. These results demonstrate that peptides identical to soybean lunasin are absent in seeds of diverse plants examined in this study.


Oxidative Stress Levels and DNA Repair Kinetics in Senescent Primary Human Fibroblasts Exposed to Chronic Low Dose Rate of Ionizing Radiation.
Exposure to low dose rate (LDR) radiation may accelerate aging processes. Previously, we identified numerous LDR-induced pathways involved in oxidative stress (OS) and antioxidant systems, suggesting that these pathways protect against premature senescence (PS). This study aimed to investigate if there are differences between young replicative senescent (RS) and PS cells considering DNA repair kinetics, OS, and DNA damage localized in the telomeres. We established PS cells by culturing and passaging young primary fibroblasts exposed to LDR. Then, RS cells were established by culturing and passaging young fibroblasts until they stopped proliferating. Senescence was characterized by analyzing telomere length and senescence-associated β-galactosidase (SA-β-gal) staining. DNA damage and repair were evaluated with γH2AX foci formation; telomere identification was carried out using the fluorescence in situ hybridization (FISH) probe; and oxidative stress was assessed by measuring 8-oxo-dG in the medium. The data indicate the following: young cells have a better ability to cope with LDR-induced oxidative stress; RS and PS have higher steady-state levels of DNA damage; RS have slower DNA repair kinetics; and PS/RS have elevated levels of telomeric DNA damage. Our main conclusion is that PS and RS differ regarding DNA repair kinetics and SA-β-gal levels.


Rapid dry exfoliation method for tuneable production of molybdenum disulphide quantum dots and large micron-dimension sheets.
Despite advances in two-dimensional (2D) transition metal dichalcogenide research owing to their outstanding physical properties, the synthesis of large micron-dimension single-layer sheets of these materials remains a challenge. Here, we present a novel and unique method to rapidly and flexibly exfoliate bulk molybdenum disulphide (MoS2) into either small nanometre-dimension quantum dots (QDs) or large micron-dimension sheets comprising predominantly single- or few-layers. The exfoliation process is conducted in dry phase, i.e., without liquid, by exploiting nanometer-amplitude MHz-order surface vibrations in the form of surface acoustic waves (SAWs). To produce the small QDs, we take advantage of the unprecedentedly large surface acceleration-on the order of 108 m s-2-to induce an iterative impaction mechanism involving successive ejection and collision of the bulk MoS2 aggregates within a miniature enclosure in order to progressively thin and break their lateral dimensions into single- and few-layer QDs. In contrast, we suppress the impaction in the zero-height enclosure limit by confining the bulk MoS2 under adhesive tape such that the shear that arises due to the travelling SAW on the substrate progressively thins the material whilst preserving their lateral dimension such that large, predominantly-monolayer, micron-sized sheets are produced with high substrate coverage up to around 80%. This fast, additive-free and dry exfoliation platform potentially presents a simple yet scalable micromechanical exfoliation method towards viable commercial production of 2D transition metal dichalcogenides.


Regression of hormone-dependent mammary tumors in Sprague-Dawley rats as a result of tamoxifen or pharmacologic doses of 17 beta-estradiol: cyclic adenosine 3',5'-monophosphate-mediated events.
The estrogen and cyclic adenosine 3',5'-monophosphate (cAMP)-binding activities changed markedly when 7,12-dimethylbenz[a]anthracene-induced mammary tumors of Sprague-Dawley rats regressed following daily injections of either tamoxifen or pharmacologic doses of 17 beta-estradiol. cAMP binding increased eightfold to tenfold, whereas estrogen binding increased twofold to threefold in regressing tumor nuclei at 5 days after either treatment, which resulted in an inversion of the ratio of estrogen binding to cAMP binding found in growing tumor nuclei. Concomitantly, both binding activities were depleted from the cytosol. In the regressing tumors, the cAMP level increased twofold and nuclear cAMP-dependent protein kinase activity increased threefold to fourfold, with a 70-80% decrease in the cytoplasmic protein kinase activity. The rise in the nuclear protein kinase activity was abolished when cycloheximide was given with tamoxifen or with high doses of 17 beta-estradiol, which suggests that the increased activity is due to new protein synthesis. In the regressing tumor nuclei, the phosphorylation of the regression-associated proteins increased, whereas the phosphorylation of growth-associated proteins decreased. These data suggest that the mammary tumor regression induced by tamoxifen or high doses of estrogen proceed through a series of cAMP-mediated events.


A new scheme for fixed node diffusion quantum Monte Carlo with pseudopotentials: Improving reproducibility and reducing the trial-wave-function bias.
Fixed node diffusion quantum Monte Carlo (FN-DMC) is an increasingly used computational approach for investigating the electronic structure of molecules, solids, and surfaces with controllable accuracy. It stands out among equally accurate electronic structure approaches for its favorable cubic scaling with system size, which often makes FN-DMC the only computationally affordable high-quality method in large condensed phase systems with more than 100 atoms. In such systems, FN-DMC deploys pseudopotentials (PPs) to substantially improve efficiency. In order to deal with nonlocal terms of PPs, the FN-DMC algorithm must use an additional approximation, leading to the so-called localization error. However, the two available approximations, the locality approximation (LA) and the T-move approximation (TM), have certain disadvantages and can make DMC calculations difficult to reproduce. Here, we introduce a third approach, called the determinant localization approximation (DLA). DLA eliminates reproducibility issues and systematically provides good quality results and stable simulations that are slightly more efficient than LA and TM. When calculating energy differences-such as interaction and ionization energies-DLA is also more accurate than the LA and TM approaches. We believe that DLA paves the way to the automation of FN-DMC and its much easier application in large systems.


Abundance and distribution of Tursiops truncatus in the Western Mediterranean Sea: an assessment towards the Marine Strategy Framework Directive requirements.
The Mediterranean Sea common bottlenose dolphin population has been assessed as Vulnerable according to the IUCN Red List Criteria. The species is also included in several International Agreements, European Union Regulations and Directives. Amongst them, a strict protection and identification of special conservation areas are requested by the EU Habitats Directive. Despite direct takes, by-catch, chemical and acoustic pollution, and prey depletion, general habitat degradation and fragmentation have been indicated as detrimental for the species, the degree to which these threats pose population risk is still largely unknown. At present it is thus not possible to depict the actual status of the population and to assess prospective trends. To address this gap in the current knowledge, line transect distance sampling aerial surveys were conducted in a wide portion of the Western Mediterranean Sea between the summer of 2010 and winter 2011. A total of 165 parallel transects equally spaced at 15 km were designed providing homogeneous coverage probability. Overall, 21,090 km were flown on effort and 16 bottlenose dolphin sightings were recorded and used for the analysis. The surface abundance and density estimates resulted in 1676 animals (CV = 38.25; 95% CI = 804-3492) with a density of 0.005 (CV = 38.25%). These results represent the first ever estimates for the common bottlenose dolphin over a wide portion of the Western Mediterranean Sea Subregion, with the potential to be useful baseline data to inform conservation. Specifically, they could be used as indicators under the Marine Strategy Framework Directive requirements, in conjunction with other study methods.


Competitive effects of intestinal microflora on Vibrio cholerae in gnotobiotic mice.
The coexistence of Vibrio cholerae and several intestinal bacteria was determined in gnotobiotic mice. The bacteria tested included a Bacteroides sp, Clostridium difficile, Clostridium perfringens type A, 2 separate isolates of Escherichia coli, 2 different Lactobacilli, 2 separate isolates of Proteus mirabilis, Pseudomonas maltophilia, and Streptococcus faecalis. Each species of bacteria became established and was recovered repeatedly from the stools during the studies. No single strain or species was found that would eliminate V cholerae from these mice within 2 wk. Not until a combination of E coli, P mirabilis, and S faecalis was present in the mice did V cholerae disappear from the stools in 14 da.


Should the label "schizophrenia" be abandoned?
The term schizophrenia is increasingly contested by researchers, clinicians, patients and families. Mental health users and professionals around the world have started calling for a change of the name, seeing it as stigmatizing and harmful. This paper reviews the literature published so far on the issue of renaming schizophrenia, carefully weighing the pros and cons of the proposed changes. Forty seven papers have been published so far, encompassing editorials, research papers, commentaries to editorials, letters, forum papers and narrative reviews. The advantages of renaming schizophrenia far outweigh the disadvantages. It would reduce stigma and benefit communication between clinicians, patients and families. The most conservative option for renaming schizophrenia would be the use of eponyms since they are neutral and avoid adverse connotations. Renaming schizophrenia is not only a matter of semantics, but also an attempt to change the stigma carried by the present name. Nevertheless, a change will not be useful unless accompanied by parallel changes in legislation, services and the education of professionals and the public.


A switch of chlorinated substrate causes emergence of a previously undetected native Dehalobacter population in an established Dehalococcoides-dominated chloroethene-dechlorinating enrichment culture.
Chlorobenzenes are soil and groundwater pollutants of concern that can be reductively dehalogenated by organohalide-respiring bacteria from the genera Dehalococcoides and Dehalobacter. The bioaugmentation culture KB-1® harbours Dehalococcoides mccartyi spp. that reductively dehalogenate trichloroethene to ethene. It contains more than 30 reductive dehalogenase genes; some of them are highly similar to genes found in the chlorobenzene-respiring Dehalococcoides mccartyi strain CBDB1. We explored the chlorobenzene dehalogenation capability of the KB-1 enrichment culture using 1,2,4-trichlorobenzene (1,2,4-TCB). We achieved adaptation of KB-1 to 1,2,4-TCB that is dehalogenated to a mixture of dichlorobenzenes, and subsequently to monochlorobenzene and benzene. Surprisingly, a native Dehalobacter population, and not a Dehalococcoides population, couples the dechlorination of 1,2,4-TCB to growth achieving an average yield of 1.1 ± 0.6 × 1013 cells per mole of Cl- released. Interestingly, the dechlorination of 1,2,4-TCB occurs alongside the complete dechlorination of trichloroethene to ethene in cultures fed both electron acceptors. Dehalobacter was not previously identified as a major player in KB-1, but its ecological niche was favoured by the introduction of 1,2,4-TCB. Based on 16S rRNA phylogeny, Dehalobacter populations seem to cluster into specialised clades, and are likely undergoing substrate specialisation as a strategy to reduce competition for electron acceptors.


A 3D Microscaffold Cochlear Electrode Array for Steroid Elution.
In cochlear implants, the electrode insertion trauma during surgery can cause damage residual hearing. Preserving the residual hearing is an important challenge and the localized administration of drugs, such as steroids, is one of the most promising ways, but remains a challenge. Here, a microscaffold cochlear electrode array (MiSCEA) consisting of a microfabricated flexible electrode array and a 3D microscaffold for steroid reservoir is reported. The MiSCEA without loaded drug is tested by measuring the electrically evoked auditory brainstem response of the cochlea in guinea pigs (n = 4). The scaffold is then coated with steroid (dexamethasone) encapsulated in polylactic-co-glycolic acid and the continuous release of the steroid into artificial perilymph during six weeks is monitored. The steroid-containing scaffolds are then implanted into guinea pigs (n = 4) and threshold shifts are analyzed for four weeks by measuring the acoustically evoked auditory brainstem response. The threshold shifts tend to be lower in the group implanted with the steroid-containing MiSCEAs. The feasibility of 3D MiSCEA opens up the development of potential next-generation cochlear electrode with improved steroid release dynamics into cochlea.


Adherence of Helicobacter pylori to human gastric epithelial cells in vitro.
Gram-negative spiral organisms, currently referred to as Helicobacter pylori, are associated with primary gastritis and duodenal ulceration. The organisms colonise gastric mucus and adhere to epithelial cells of inflamed antra. To further examine the binding of H. pylori to human gastric epithelial cells, we developed and characterised an in-vitro bacterial adherence assay. Scanning electronmicroscopy suggested that spiral-shaped bacteria were adherent to the surface of KATO-III cells which were derived from a human gastric adenocarcinoma. Transmission electronmicroscopy confirmed the attachment of H. pylori to these epithelial cells in tissue culture. Some bacteria were adherent to intact microvilli, others were closely adherent to the plasma membrane in regions where microvilli were effaced. In studies with radiolabelled H. pylori, adherence to epithelial cells in tissue culture contrasted with minimal binding of bacteria to polystyrene wells alone. Incubation of bacteria with gastric cells at 4 degrees C significantly reduced adherence of H. pylori. We conclude that adherence of H. pylori to gastric epithelial cells in tissue culture involved "attachment and effacement mechanisms". This assay could serve as a suitable in-vitro model for the study of the bacterial adhesins and host receptors which mediate attachment of H. pylori to gastric epithelial cell surfaces.


Myofibroblast-Derived Exosome Induce Cardiac Endothelial Cell Dysfunction.
Background: Endothelial cells (ECs) play a critical role in the maintenance of vascular homeostasis and in heart function. It was shown that activated fibroblast-derived exosomes impair cardiomyocyte function in hypertrophic heart, but their effect on ECs is not yet clear. Thus, we hypothesized that activated cardiac fibroblast-derived exosomes (FB-Exo) mediate EC dysfunction, and therefore modulation of FB-exosomal contents may improve endothelial function. Methods and Results: Exosomes were isolated from cardiac fibroblast (FB)-conditioned media and characterized by nanoparticle tracking analysis and electron microscopy. ECs were isolated from mouse heart. ECs were treated with exosomes isolated from FB-conditioned media, following FB culture with TGF-β1 (TGF-β1-FB-Exo) or PBS (control) treatment. TGF-β1 significantly activated fibroblasts as shown by increase in collagen type1 α1 (COL1α1), periostin (POSTN), and fibronectin (FN1) gene expression and increase in Smad2/3 and p38 phosphorylation. Impaired endothelial cell function (as characterized by a decrease in tube formation and cell migration along with reduced VEGF-A, Hif1α, CD31, and angiopoietin1 gene expression) was observed in TGF-β1-FB-Exo treated cells. Furthermore, TGF-β1-FB-Exo treated ECs showed reduced cell proliferation and increased apoptosis as compared to control cells. TGF-β1-FB-Exo cargo analysis revealed an alteration in fibrosis-associated miRNAs, including a significant increase in miR-200a-3p level. Interestingly, miR-200a-3p inhibition in activated FBs, alleviated TGF-β1-FB-Exo-mediated endothelial dysfunction. Conclusions: Taken together, this study demonstrates an important role of miR-200a-3p enriched within activated fibroblast-derived exosomes on endothelial cell biology and function.


The Modulatory Bioeffects of Pomegranate (Punica granatum L.) Polyphenols on Metabolic Disorders: Understanding Their Preventive Role against Metabolic Syndrome.
Modern research achievements support the health-promoting effects of natural products and diets rich in polyphenols. Pomegranate (PG) (Punica granatum L.) contains a considerable number of bioactive compounds that exert a broad spectrum of beneficial biological activities, including antimicrobial, antidiabetic, antiobesity, and atheroprotective properties. In this context, the reviewed literature shows that PG intake might reduce insulin resistance, cytokine levels, redox gene expression, blood pressure elevation, vascular injuries, and lipoprotein oxidative modifications. The lipid parameter corrective capabilities of PG-ellagitannins have also been extensively reported to be significantly effective in reducing hyperlipidemia (TC, LDL-C, VLDL-C, and TAGs), while increasing plasma HDL-C concentrations and improving the TC/HDL-C and LDL-C/HDL-C ratios. The health benefits of pomegranate consumption seem to be acheived through the amelioration of adipose tissue endocrine function, fatty acid utilization, GLUT receptor expression, paraoxonase activity enhancement, and the modulation of PPAR and NF-κB. While the results from animal experiments are promising, human findings published in this field are inconsistent and are still limited in several aspects. The present review aims to discuss and provide a critical analysis of PG's bioeffects on the components of metabolic syndrome, type-2 diabetes, obesity, and dyslipidemia, as well as on certain cardiovascular-related diseases. Additionally, a brief overview of the pharmacokinetic properties, safety, and bioavailability of PG-ellagitannins is included.


Design, Synthesis, Antiviral Evaluation, and SAR Studies of New 1-(Phenylsulfonyl)-1H-Pyrazol-4-yl-Methylaniline Derivatives.
A series of N-((3-phenyl-1-(phenylsulfonyl)-1H-pyrazol-4-yl)methyl)anilines 7a-p and 8a-l, structurally related to previously synthesized and tested (N-(1,3-diphenyl-1H-pyrazol-4-yl)methyl)anilines (1a-v), were designed and synthesized. The new derivatives were evaluated in cell-based assays for their cytotoxicity and antiviral activity against a large panel of RNA and DNA viruses of public health significance. Generally, the tested compounds did not display cytotoxicity toward the cell lines used. The majority of derivatives 7a-p were able to interfered with YFV and RSV replication in the micromolar range showing a marked improvement in potency and selectivity with respect to the reference inhibitors 6-azauridine and ribavirin, respectively. The introduction of a p-methoxy substituent on the phenylsulfonyl group (compounds 8a-l) completely abolished the anti-RSV activity and reduced or eliminated the potency against YFV. On the contrary, several p-methoxy analogs were able to interfere with BVDV replication with a comparable (8b, 8c, 8g, and 8k) or better (8a and 8f) potency than the reference inhibitor, ribavirin. Compound 7e, selected for time of addition experiments on BHK-21 cell cultures infected with YFV, achieved the highest reduction of virus titer when added 2 h post infection and maintained up to 4 h post infection.


RAS1 regulates filamentation, mating and growth at high temperature of Cryptococcus neoformans.
Cryptococcus neoformans is a basidiomycete yeast and opportunistic human pathogen of increasing clinical importance due to the increasing population of immunocompromised patients. To further investigate signal transduction cascades regulating fungal pathogenesis, we have identified the gene encoding a RAS homologue in this organism. The RAS1 gene was disrupted by transformation and homologous recombination. The resulting ras1 mutant strain was viable, but failed to grow at 37 degrees C, and exhibited significant defects in mating and agar adherence. The ras1 mutant strain was also avirulent in an animal model of cryptococcal meningitis. Reintroduction of the wild-type RAS1 gene complemented these ras1 mutant phenotypes and restored virulence in animals. A dominantly active RAS1 mutant allele, RAS1Q67L, induced a differentiation phenotype known as haploid fruiting, which involves filamentation, agar invasion and sporulation in response to nitrogen deprivation. The ras1 mutant mating defect was suppressed by overexpression of MAP kinase signalling elements and partially suppressed by exogenous cAMP. Additionally, cAMP also suppressed the agar adherence defect of the ras1 mutant. However, the ability of the ras1 mutant strain to grow at elevated temperature was not restored by cAMP or MAP kinase overexpression. Our findings support a model in which RAS1 signals in C. neoformans through cAMP-dependent, MAP kinase, and RAS-specific signalling cascades to regulate mating and filamentation, as well as growth at high temperature which is necessary for maintenance of infection.


Infrared study of the kinetics and mechanism of adsorption of acrylic polymers on alumina surfaces.
In this paper, we studied the kinetics of the adsorption of poly(methyl methacrylate), PMMA, onto native aluminum oxide surfaces by X-ray photoelectron spectroscopy and reflection-absorption infrared spectroscopy, with the intent of tracking the various changes observed in the infrared spectrum of the adsorbed polymer layer as a function of adsorption time. Specifically, we utilized the relative changes in the absorption bands of the carbonyl, carboxylic acid, and carboxylate groups to determine the sequence of events that culminate in the formation of bonds between carboxylate groups on hydrolyzed PMMA and specific sites on the aluminum oxide surface. We have shown that the adsorption process involves the hydrolysis of a fraction of the methoxy groups of the PMMA to generate COOH groups. Unlike previous assumptions, the formation of COOH groups on the PMMA chains does not constitute a sufficient condition for the actual chemisorption of the polymer chains onto the metal oxide surface. To promote bonding, the acid groups must undergo dissociation to form the carboxylate groups, followed subsequently by actual bond formation, that is, active anchoring, on the surface. This process is mediated by the aluminum oxide sites on the surface in the presence of water. Hence, the adsorption process occurs via a two-step mechanism, in which the first step, that is, the hydrolysis step, is a necessary but insufficient condition and the second step, that is, the anchoring step, is largely dependent on the type of interfacial chemistry possible for a particular polymer-metal oxide surface, the polymer conformation, the molecular weight, and the flexibility of the adsorbing molecules.


Nitrogen losses and greenhouse gas emissions under different N and water management in a subtropical double-season rice cropping system.
Nitrogen non-point pollution and greenhouse gas (GHG) emission are major challenges in rice production. This study examined options for both economic and environmental sustainability through optimizing water and N management. Field experiments were conducted to examine the crop yields, N use efficiency (NUE), greenhouse gas emissions, N losses under different N and water management. There were four treatments: zero N input with farmer's water management (N0), farmer's N and water management (FP), optimized N management with farmer's water management (OPTN) and optimized N management with alternate wetting and drying irrigation (OPTN+AWD). Grain yields in OPTN and OPTN+AWD treatments increased by 13.0-17.3% compared with FP. Ammonia volatilization (AV) was the primary pathway for N loss for all treatments and accounted for over 50% of the total losses. N losses mainly occurred before mid-tillering. N losses through AV, leaching and surface runoff in OPTN were reduced by 18.9-51.6% compared with FP. OPTN+AWD further reduced N losses from surface runoff and leaching by 39.1% and 6.2% in early rice season, and by 46.7% and 23.5% in late rice season, respectively, compared with OPTN. The CH4 emissions in OPTN+AWD were 20.4-45.4% lower than in OPTN and FP. Total global warming potential of CH4 and N2O was the lowest in OPTN+AWD. On-farm comparison confirmed that N loss through runoff in OPTN+AWD was reduced by over 40% as compared with FP. OPTN and OPTN+AWD significantly increased grain yield by 6.7-13.9%. These results indicated that optimizing water and N management can be a simple and effective approach for enhancing yield with reduced environmental footprints.


Sonic hedgehog (Shh) signaling promotes tumorigenicity and stemness via activation of epithelial-to-mesenchymal transition (EMT) in bladder cancer.
Activation of the sonic hedgehog (Shh) signaling pathway controls tumorigenesis in a variety of cancers. Here, we show a role for Shh signaling in the promotion of epithelial-to-mesenchymal transition (EMT), tumorigenicity, and stemness in the bladder cancer. EMT induction was assessed by the decreased expression of E-cadherin and ZO-1 and increased expression of N-cadherin. The induced EMT was associated with increased cell motility, invasiveness, and clonogenicity. These progression relevant behaviors were attenuated by treatment with Hh inhibitors cyclopamine and GDC-0449, and after knockdown by Shh-siRNA, and led to reversal of the EMT phenotype. The results with HTB-9 were confirmed using a second bladder cancer cell line, BFTC905 (DM). In a xenograft mouse model TGF-β1 treated HTB-9 cells exhibited enhanced tumor growth. Although normal bladder epithelial cells could also undergo EMT and upregulate Shh with TGF-β1 they did not exhibit tumorigenicity. The TGF-β1 treated HTB-9 xenografts showed strong evidence for a switch to a more stem cell like phenotype, with functional activation of CD133, Sox2, Nanog, and Oct4. The bladder cancer specific stem cell markers CK5 and CK14 were upregulated in the TGF-β1 treated xenograft tumor samples, while CD44 remained unchanged in both treated and untreated tumors. Immunohistochemical analysis of 22 primary human bladder tumors indicated that Shh expression was positively correlated with tumor grade and stage. Elevated expression of Ki-67, Shh, Gli2, and N-cadherin were observed in the high grade and stage human bladder tumor samples, and conversely, the downregulation of these genes were observed in the low grade and stage tumor samples. Collectively, this study indicates that TGF-β1-induced Shh may regulate EMT and tumorigenicity in bladder cancer. Our studies reveal that the TGF-β1 induction of EMT and Shh is cell type context dependent. Thus, targeting the Shh pathway could be clinically beneficial in the ability to reverse the EMT phenotype of tumor cells and potentially inhibit bladder cancer progression and metastasis.


Minimally invasive management of epiphrenic esophageal diverticula.
The purpose of this study is to review our initial experience with a minimally invasive approach to manage symptomatic epiphrenic esophageal diverticula. Five patients with symptomatic epiphrenic esophageal diverticula underwent surgical management between August 1997 and December 2002. All patients complained of dysphagia; had experienced symptoms for at least 12 months; and were evaluated preoperatively by a barium esophagram, esophagogastroduodenoscopy, and esophageal manometry. The epiphrenic esophageal diverticula measured 5 cm or less in all patients. Manometry demonstrated esophageal dysmotility in three patients. A minimally invasive technique was completed in all five patients. Four patients underwent laparoscopic diverticulectomy and myotomy including a concomitant Toupet fundoplication, and one patient underwent thoracoscopic diverticulectomy and myotomy. The mean operative time was 245 minutes (range 175-334). The longest operative time was for the thoracoscopic procedure. The estimated blood loss was minimal (range 30-100 cm3). The laparoscopic patients had a mean postoperative length of stay of 2.75 days (range 2-4) and the patient undergoing a thoracoscopic approach was discharged on postoperative day 6 due to a history of lung disease and home oxygen requirements. There were no other postoperative complications. After a mean follow-up of 16.2 months (range 3-36) all patients are asymptomatic. Short-term follow-up after our initial experience with minimally invasive approaches for epiphrenic esophageal diverticula demonstrates that thoracoscopic and laparoscopic approaches are feasible; safe; and effectively alleviate dysphagia, regurgitation, and other associated symptoms. Long-term outcomes should be monitored during the evolution of these novel minimally invasive techniques to ensure outcomes comparable to those of a transthoracic open approach.


Prostate-specific antigen testing for prostate cancer screening: A national survey of Canadian primary care physicians' opinions and practices.
In 2014, the Canadian Task Force on Preventive Health Care (CTFPHC) recommended against routine prostate cancer screening with the prostate-specific antigen (PSA) blood test. We surveyed Canadian primary care physicians (PCPs) to understand their opinions and attitudes towards prostate cancer screening in 2016. Twenty PCPs piloted the survey to assess its accessibility. We distributed a flyer to 19 633 PCPs as an insert in a large mailed package inviting them to attend a national meeting, and later promoted the survey at the meeting. Multinomial logistic regression models examined factors associated with agreement of key guideline statements and the overall benefit of PSA screening. A total of 1254 PCPs responded (rate of 6.4%); 54.7% of physicians aware of the CTFPHC recommendations report screening less often as a result. Overall, 55.6% of PCPs feel that the risks of PSA screening outweigh the benefits. On multivariable analysis, physicians who did not read the guidelines, did not have an academic appointment, or were in practice for over 20 years were significantly more likely to disagree with the statement that men 55-69 years old should not be screened for prostate cancer with PSA. Our national survey found that the prostate cancer screening practices of Canadian PCPs varies widely across physician demographic groups, with almost equal numbers for or against. This has significant ethical, medical, and legal implications. The poor response rate to highly incentivized survey request may suggest a reluctance or general apathy towards this subject because of the Task Force recommendations. Future efforts should provide physicians with objective guidance around PSA screening, incorporating input from all stakeholders, including PCPs, urologists, and patients.


The association between depression and eating styles in four European countries: The MooDFOOD prevention study.
Depression, one of the most prevalent and disabling disorders in Europe, is thought to be associated with unhealthy eating styles. As prevalence of depression and eating styles potentially differ across Europe, the current study aimed to investigate in a large, European sample, the associations of history of major depressive disorder and depression severity with unhealthy eating styles. Baseline data of the MooDFOOD prevention study was used. The current analysis included 990 participants of four European countries (The Netherlands, United Kingdom, Germany, Spain). Analyses of Covariance and linear regression analyses were performed with depression history or depression severity as determinants, and emotional, uncontrolled, and cognitive restrained eating (Three Factor Eating Questionnaire Revised, 18 item) as outcomes. Depression history and severity were associated with more emotional and uncontrolled eating and with less cognitive restrained eating. Mood, somatic, and cognitive symptom clusters were also associated with more emotional and uncontrolled eating, and with less cognitive restrained eating. The somatic depressive symptoms "increased appetite" and "increased weight" were more strongly associated to unhealthy eating styles compared to other symptoms. No differences in associations between depression and unhealthy eating were found between European countries. Our results suggest that depression is related to more unhealthy eating styles. Diminishing unhealthy eating styles in subthreshold depressed persons could potentially reduce adverse health consequences like weight gain, unhealthy dietary patterns and weight-related diseases. It is also possible that interventions that decrease depressive symptoms can lead to a decrease in unhealthy eating styles.


Sequence variants in the transforming growth beta-induced factor (TGIF) gene are not associated with high myopia.
High myopia is a common complex-trait eye disorder, with implications for blindness due to increased risk of retinal detachment, macular degeneration, premature cataracts, and glaucoma. Mapping studies have identified at least four loci for nonsyndromic autosomal dominant high myopia at 18p11.31, 12q22-q23, 17q21-q23, and 7q36. The smallest haplotyped interval for these loci is that of the MYP2 locus on 18p11.31. Recently, the transforming growth beta-induced factor (TGIF) gene was reported to be a candidate gene for MYP2-associated high myopia in single-nucleotide polymorphism studies. The purpose of this study was to determine whether DNA sequence variants in the human TGIF gene are causally related to MYP2-associated high myopia. The protein coding regions and intron-exon boundaries of the human TGIF gene were sequenced using genomic DNA samples from MYP2 individuals (affected, unaffected) and external control subjects. The TGIF model used was the April 20, 2003, human genome National Center for Biotechnology Information (NCBI) build 33, which has 10 exons and encodes eight transcript variants. Polymorphic sequence changes were compared to those in the previous report. Reverse-transcription polymerase chain reaction (RT-PCR) was performed to validate TGIF gene expression in ocular tissues. A total of 21 polymorphisms of TGIF were found by direct sequencing: 3 were missense, 2 were silent, 10 were not translated, 4 were intronic, and 2 were homozygous deletions. The 3 missense allelic variants were localized to exon 10 at positions 236C-->T(Pro-->Leu), 244C-->T(Pro-->Ser), and 245C-->T(Pro-->Leu). Silent mutations were observed in exon 10 at positions 177A-->G, 333C-->T. Ten polymorphisms were novel. No sequence alterations were exclusively associated with the affected disease phenotype. RT-PCR results confirmed expression of TGIF in RNA samples derived from human sclera, cornea, optic nerve, and retina. TGIF is a known candidate gene for MYP2-associated high myopia, based on its mapped location within the MYP2 interval. Mutation analysis of the encoded TGIF gene for MYP2 autosomal dominant high myopia did not identify sequence alterations associated with the disease phenotype. Further studies of MYP2 candidate genes are needed to determine the gene that causes of this potentially blinding disorder.


Dendritic cell vaccines and immunity in glioma patients.
The dismal prognoses suffered by malignant primary brain tumor (glioma) patients remain unchanged over the past two decades despite significant improvements in the treatment of distinct tumors. Immunotherapy, and vaccine therapy in particular, represents a promising experimental approach to treat malignant gliomas, but major challenges still remain to render vaccination clinically effective. These challenges include diminishing the risk of pathologic autoimmunity, and identifying the cellular basis of clinical vaccine benefits. Addressing such challenges should eventually help increase the proportion of patients experiencing clinical vaccine benefits. Recent studies in glioma patients have characterized tumor antigens on human gliomas, identified some of the immune cells involved in beneficial anti-glioma immunity, and examined how gliomas may be altered by sub-lethal immune influences. This has provided a glimpse of the strength to which immunity influences glioma clinical outcome, and resurrects hope that clinically effective vaccines to treat these tumors is within reach. Insight into the complex dynamics of immune-tumor interactions promises to extend this reach by delineating mechanisms of immune synergy with other forms of treatment.