Extracellular waveforms reveal an axonal origin of spikelets in pyramidal neurons.
Spikelets are small spike-like membrane depolarizations measured at the soma whose origin in pyramidal neurons is still unresolved. We investigated the mechanism of spikelet generation using detailed models of pyramidal neurons. We simulated extracellular waveforms associated with action potentials and spikelets and compared these with experimental data obtained by Chorev and Brecht ( J Neurophysiol 108: 1584-1593, 2012) from hippocampal pyramidal neurons in vivo. We considered spikelets originating in the axon of a single cell as well as spikelets generated in two cells coupled with gap junctions. We found that in both cases the experimental data can be explained by an axonal origin of spikelets: in the single-cell case, action potentials are generated in the axon but fail to activate the soma. Such spikelets can be evoked by dendritic input. Alternatively, spikelets resulting from axoaxonal gap junction coupling with a large (greater than several hundred μm) distance between the somata of the coupled cells are also consistent with the data. Our results demonstrate that a cell firing a somatic spikelet generates a detectable extracellular potential that is different from the action potential-correlated extracellular waveform generated by the same cell and recorded at the same location. This, together with the absence of a refractory period between action potentials and spikelets, implies that spikelets and action potentials generated in one cell may easily get misclassified in extracellular recordings as two different cells, albeit they both constitute the output of a single pyramidal neuron. NEW & NOTEWORTHY We addressed the origin of spikelets, using compartmental models of pyramidal neurons. Comparing our simulation results with published extracellular spikelet recordings revealed an axonal origin of spikelets. Our results imply that action potential- and spikelet-associated extracellular waveforms may easily get misclassified as two different cells, albeit they both constitute the output of a single pyramidal cell.


Labeling of fusion proteins with synthetic fluorophores in live cells.
A general approach for the sequential labeling of fusion proteins of O(6)-alkylguanine-DNA alkyltransferase (AGT) with different fluorophores in mammalian cells is presented. AGT fusion proteins with different localizations in the cell can be labeled specifically with different fluorophores, and the fluorescence labeling can be used for applications such as multicolor analysis of dynamic processes and fluorescence resonance energy transfer measurements. The facile access to a variety of different AGT substrates as well as the specificity of the labeling reaction should make the approach an important tool to study protein function in live cells.


Trace element concentrations along a gradient of urban pressure in forest and lawn soils of the Paris region (France).
The concentration, degree of contamination and pollution of 7 trace elements (TEs) along an urban pressure gradient were measured in 180 lawn and wood soils of the Paris region (France). Iron (Fe), a major element, was used as reference element. Copper (Cu), cadmium (Cd), lead (Pb) and zinc (Zn) were of anthropogenic origin, while arsenic (As), chromium (Cr) and nickel (Ni) were of natural origin. Road traffic was identified as the main source of anthropogenic TEs. In addition, the industrial activity of the Paris region, especially cement plants, was identified as secondary source of Cd. Soil characteristics (such as texture, organic carbon (OC) and total nitrogen (tot N) contents) tell the story of the soil origins and legacies along the urban pressure gradient and often can explain TE concentrations. The history of the land-use types was identified as a factor that allowed understanding the contamination and pollution by TEs. Urban wood soils were found to be more contaminated and polluted than urban lawns, probably because woods are much older than lawns and because of the legacy of the historical management of soils in the Paris region (Haussmann period). Lawn soils are similar to the fertile agricultural soils and relatively recently (mostly from the 1950s onwards) imported from the surrounding of Paris, so that they may be less influenced by urban conditions in terms of TE concentrations. Urban wood soils are heavily polluted by Cd, posing a high risk to the biological communities. The concentration of anthropogenic TEs increased from the rural to the urban areas, and the concentrations of most anthropogenic TEs in urban areas were equivalent to or above the regulatory reference values, raising the question of longer-term monitoring.


Transmission dynamics of Tasmanian devil facial tumor disease may lead to disease-induced extinction.
Most pathogens threatening to cause extinction of a host species are maintained on one or more reservoir hosts, in addition to the species that is threatened by disease. Further, most conventional host-pathogen theory assumes that transmission is related to host density, and therefore a pathogen should become extinct before its sole host. Tasmanian devil facial tumor disease is a recently emerged infectious cancer that has led to massive population declines and grave concerns for the future persistence of this largest surviving marsupial carnivore. Here we report the results of mark-recapture studies at six sites and use these data to estimate epidemiological parameters critical to both accurately assessing the risk of extinction from this disease and effectively managing this disease threat. Three sites were monitored from before or close to the time of disease arrival, and at three others disease was well established when trapping began, in one site for at least 10 years. We found no evidence for sex-specific differences in disease prevalence and little evidence of consistent seasonal variation in the force of infection. At all sites, the disease was maintained at high levels of prevalence (>50% in 2-3-year-old animals), despite causing major population declines. We also provide the first estimates of the basic reproductive rate R0 for this disease. Using a simple age-structured deterministic model, we show that our results are not consistent with transmission being proportional to the density of infected hosts but are consistent with frequency-dependent transmission. This conclusion is further supported by the observation that local disease prevalence in 2-3-year-olds still exceeds 50% at a site where population density has been reduced by up to 90% in the past 12 years. These findings lend considerable weight to concerns that this host-specific pathogen will cause the extinction of the Tasmanian devil. Our study highlights the importance of rapidly implementing monitoring programs to determine how transmission depends on host density and emphasizes the need for ongoing management strategies involving a disease-free "insurance population," along with ongoing field monitoring programs to confirm whether local population extinction occurs.


Improving axial resolution in confocal microscopy with new high refractive index mounting media.
Resolution, high signal intensity and elevated signal to noise ratio (SNR) are key issues for biologists who aim at studying the localisation of biological structures at the cellular and subcellular levels using confocal microscopy. The resolution required to separate sub-cellular biological structures is often near to the resolving power of the microscope. When optimally used, confocal microscopes may reach resolutions of 180 nm laterally and 500 nm axially, however, axial resolution in depth is often impaired by spherical aberration that may occur due to refractive index mismatches. Spherical aberration results in broadening of the point-spread function (PSF), a decrease in peak signal intensity when imaging in depth and a focal shift that leads to the distortion of the image along the z-axis and thus in a scaling error. In this study, we use the novel mounting medium CFM3 (Citifluor Ltd., UK) with a refractive index of 1.518 to minimize the effects of spherical aberration. This mounting medium is compatible with most common fluorochromes and fluorescent proteins. We compare its performance with established mounting media, harbouring refractive indices below 1.500, by estimating lateral and axial resolution with sub-resolution fluorescent beads. We show furthermore that the use of the high refractive index media renders the tissue transparent and improves considerably the axial resolution and imaging depth in immuno-labelled or fluorescent protein labelled fixed mouse brain tissue. We thus propose to use those novel high refractive index mounting media, whenever optimal axial resolution is required.


Alterations in vasomotor control of coronary resistance vessels in remodelled myocardium of swine with a recent myocardial infarction.
The mechanism underlying the progressive deterioration of left ventricular (LV) dysfunction after myocardial infarction (MI) towards overt heart failure remains incompletely understood, but may involve impairments in coronary blood flow regulation within remodelled myocardium leading to intermittent myocardial ischemia. Blood flow to the remodelled myocardium is hampered as the coronary vasculature does not grow commensurate with the increase in LV mass and because extravascular compression of the coronary vasculature is increased. In addition to these factors, an increase in coronary vasomotor tone, secondary to neurohumoral activation and endothelial dysfunction, could also contribute to the impaired myocardial oxygen supply. Consequently, we explored, in a series of studies, the alterations in regulation of coronary resistance vessel tone in remodelled myocardium of swine with a 2 to 3-week-old MI. These studies indicate that myocardial oxygen balance is perturbed in remodelled myocardium, thereby forcing the myocardium to increase its oxygen extraction. These perturbations do not appear to be the result of blunted beta-adrenergic or endothelial NO-mediated coronary vasodilator influences, and are opposed by an increased vasodilator influence through opening of K(ATP) channels. Unexpectedly, we observed that despite increased circulating levels of noradrenaline, angiotensin II and endothelin-1, alpha-adrenergic tone remained negligible, while the coronary vasoconstrictor influences of endogenous endothelin and angiotensin II were virtually abolished. We conclude that, early after MI, perturbations in myocardial oxygen balance are observed in remodelled myocardium. However, adaptive alterations in coronary resistance vessel control, consisting of increased vasodilator influences in conjunction with blunted vasoconstrictor influences, act to minimize the impairments of myocardial oxygen balance.


Abiotic Stress Responses and Microbe-Mediated Mitigation in Plants: The Omics Strategies.
Abiotic stresses are the foremost limiting factors for agricultural productivity. Crop plants need to cope up adverse external pressure created by environmental and edaphic conditions with their intrinsic biological mechanisms, failing which their growth, development, and productivity suffer. Microorganisms, the most natural inhabitants of diverse environments exhibit enormous metabolic capabilities to mitigate abiotic stresses. Since microbial interactions with plants are an integral part of the living ecosystem, they are believed to be the natural partners that modulate local and systemic mechanisms in plants to offer defense under adverse external conditions. Plant-microbe interactions comprise complex mechanisms within the plant cellular system. Biochemical, molecular and physiological studies are paving the way in understanding the complex but integrated cellular processes. Under the continuous pressure of increasing climatic alterations, it now becomes more imperative to define and interpret plant-microbe relationships in terms of protection against abiotic stresses. At the same time, it also becomes essential to generate deeper insights into the stress-mitigating mechanisms in crop plants for their translation in higher productivity. Multi-omics approaches comprising genomics, transcriptomics, proteomics, metabolomics and phenomics integrate studies on the interaction of plants with microbes and their external environment and generate multi-layered information that can answer what is happening in real-time within the cells. Integration, analysis and decipherization of the big-data can lead to a massive outcome that has significant chance for implementation in the fields. This review summarizes abiotic stresses responses in plants in-terms of biochemical and molecular mechanisms followed by the microbe-mediated stress mitigation phenomenon. We describe the role of multi-omics approaches in generating multi-pronged information to provide a better understanding of plant-microbe interactions that modulate cellular mechanisms in plants under extreme external conditions and help to optimize abiotic stresses. Vigilant amalgamation of these high-throughput approaches supports a higher level of knowledge generation about root-level mechanisms involved in the alleviation of abiotic stresses in organisms.


In vivo, high-resolution analysis of yeast and mammalian RNA-protein interactions, RNA structure, RNA splicing and ribozyme cleavage by use of terminal transferase-dependent PCR.
We have investigated the analysis of RNA by use of terminal transferase-dependent PCR (TDPCR), a procedure previously used for the analysis of DNA and chromatin [J. Komura and A.D.Riggs, Nucleic Acids Res.,26, 1807-1811 (1998)]. When preceded by reverse transcription (RT), TDPCR provides an extremely sensitive, versatile, quantitative and nucleotide-level assay for detecting RNA lesions or structures that block primer extension during the RT step. The procedure is: (i) RT using a gene-specific oligonucleotide; (ii) ribo-tailing of the single-stranded cDNA product by use of terminal deoxy-nucleotidyl transferase; (iii) ligation of a DNA linker to the tailed cDNA by use of T4 DNA ligase; and (iv) PCR using a nested, gene-specific primer and a linker-specific primer. This procedure combines the versatility of a primer extension assay with nucleotide-level resolution, the specificity of nested primers and the sensitivity of PCR. Band patterns obtained are reproducible and quantifiable. We successfully used the technique for the study of yeast RNA structure, splicing intermediates and ribozyme cleavage. Also, in vivo footprint experiments, using mammalian cells and RNase T1, revealed the binding of iron-responsive element binding protein to iron responsive elements in the mRNAs of transferrin receptor and ferritin H-chain.


Romaine Lettuce/Skullcap Mixture Improves Sleep Behavior in Vertebrate Models.
The aim of this study is to investigate the effects of romaine lettuce leaves extract (RE), skullcap root extract (SE) and their mixture on sleep behaviors in vertebrate models. HPLC analysis showed that RE contains lactucopicrin (0.02±0.01 mg/g extract), chlorogenic acid (4.05±0.03 mg/g extract), caffeic acid (2.38±0.03 mg/g extract), and chicoric acid (7.02±0.32 mg/g extract) as main phenolic compounds, while SE includes baicalin (99.4±0.5 mg/g extract), baicalein (8.28±0.21 mg/g extract), and wogonin (3.09±0.32 mg/g extract). The mixture of RE (100 mg/g extract) and SE (40 mg/g extract) increased total sleep time by 50.9% compared with the control in pentobarbital-induced sleep model. In electroencephalography (EEG) analysis, RE/SE mixture significantly increased Non-Rapid Eye Movement (NREM), in which delta wave was enhanced by around 40% compared with normal control, leading to the increase of sleep time. In caffeine-induced wake model, RE/SE mixture greatly decreased (53%) caffeine-induced wake time, showing a similar level to normal control. In addition, caffeine-induced decreased of NREM and delta wave effectively increased with RE/SE mixture; NREM and delta wave increased by 85% and 108%, respectively. Furthermore, RE/SE mixture was shown to bind to a gamma-aminobutyric acid type A (GABAA)-benzodiazepine (BZD) receptor stronger than RE or SE single extract. Taken together, RE/SE mixture effectively improved sleep behavior with the increase of NREM via GABAA-BZD receptor binding. RE/SE mixture can be used as an herbal agent for sleep disorders.


Holographic diffractive collimators based on recording with homocentric diverging beams for diode lasers.
A single collimating transmission off-axis blazed surface-relief holographic diffractive element that corrects the ellipticity and the astigmatism of a diode-laser beam was designed. The procedure was applied to a hypothetical laser operating at a wavelength of 633 nm that possesses extremely large astigmatism and relatively small ellipticity together with small diverging angles of the laser beam. The element was recorded with only homocentric diverging beams that had pure spherical wave fronts. Because of the off-axis nature of the element, the aberration of asymmetry had to be corrected. The element was tested in a reciprocal way when the incident wave was a collimated He-Ne laser beam. A diffraction-limited beam was obtained that was as large as approximately half of the designed aperture.


Oncolytic immunotherapy: unlocking the potential of viruses to help target cancer.
Oncolytic immunotherapy is a research area of cancer immunotherapy investigating the use of modified viruses to target cancer cells. A variety of different viral backbones (e.g., adenovirus, reovirus) with a diverse range of genetic modifications are currently being investigated for the treatment of a variety of cancers. The oncolytic virus that has advanced the furthest in clinical development is talimogene laherparepvec, a recombinant HSV-1 virus expressing granulocyte-macrophage colony-stimulating factor (GM-CSF). In a phase 3 study in patients with unresectable metastatic melanoma, intralesional talimogene laherparepvec treatment resulted in a higher durable response rate compared with subcutaneous GM-CSF treatment (16.3 versus 2.1%; P < 0.001). Notably, responses were observed at uninjected lesions including visceral lesions, indicating a systemic antitumor response had occurred. Studies evaluating combination treatments involving oncolytic viruses and immunologic agents are ongoing. This review focuses on the mechanisms of action for oncolytic viruses and highlights select agents and combinations currently in development.


Inhibition and activation of acetaminophen reactive metabolite formation by caffeine. Roles of cytochromes P-450IA1 and IIIA2.
Caffeine has previously been shown to diminish or potentiate acetaminophen (APAP) hepatotoxicity in rats, depending on induction state. To elucidate the P-450 forms involved in these divergent effects, rat liver microsomes, prepared after pretreatment with various inducers, were used to examine the effect of caffeine on N-acetyl-p-benzoquinone imine (NAPQI) formation. The addition of caffeine to incubations with 3-methylcholanthrene (MC)-induced microsomes resulted in a biphasic effect on the formation of NAPQI. A 43% decrease in NAPQI formation was observed as caffeine concentration was increased from 0 to 0.5 mM; however, NAPQI formation was accelerated as caffeine concentration increased, exceeding the control (no caffeine) value, at caffeine concentrations greater than 2.5 mM. Incubations with purified P-450IA1 showed that as caffeine concentration increased from 0 to 5 mM, a 50% inhibition was observed with no evidence of acceleration. In contrast to MC microsomes, the addition of caffeine to incubations with uninduced and phenobarbital-induced adult rat microsomes resulted in a marked (3- to 4-fold) acceleration of NAPQI formation with no evidence of inhibition. Caffeine (5 mM) also accelerated NAPQI formation in microsomes isolated from diabetic rats, but to a substantially lesser extent (120%), suggesting a modest (if any) effect on P-450IIE1, a form previously shown to form NAPQI from APAP. Interestingly, caffeine caused a 3- to 4-fold increase in NAPQI formation by juvenile male and female rat microsomes, but no activation was observed with adult female rat microsomes. These results suggested that caffeine activated a member of the cytochrome P-450IIIA subfamily.(ABSTRACT TRUNCATED AT 250 WORDS)


Reduced hippocampal volume and total white matter volume in posttraumatic stress disorder.
Reduced hippocampal volumes in posttraumatic stress disorder (PTSD) patients are thought to reflect specific changes of this structure. Previous magnetic resonance imaging (MRI) studies have not consistently examined indices of overall brain atrophy, therefore it cannot be completely ruled out that hippocampal changes are explained by whole-brain atrophy. The purpose of this study was to assess hippocampal and whole-brain volume in civilian PTSD. Twelve subjects with PTSD and 10 control subjects underwent brain MRI. Hippocampal volumes were visually quantified using a computerized volumetric program. Whole-brain volumes were obtained with automated k-means-based segmentation. No differences were found in intracranial volumes (ICV). Subjects with PTSD had higher cerebrospinal fluid (CSF)/ICV ratios and lower white matter/ICV ratios, consistent with generalized white matter (WM) atrophy. The effect of age on CSF/ICV was more pronounced in the PTSD group. Subjects with PTSD had smaller absolute and normalized bilateral hippocampal volumes. These differences persisted after adjusting for lifetime weeks of alcohol intoxication. Posttraumatic stress disorder and depression scores correlated negatively with left hippocampal volume, but PTSD scores were a better predictor of hippocampal volumes. Our results replicate previous findings of reduced hippocampal volume in PTSD but also suggest independent, generalized, white matter atrophy.


Using Eye Tracking to Explore Facebook Use and Associations with Facebook Addiction, Mental Well-being, and Personality.
Social networking sites (SNSs) have become ubiquitous in our everyday lives, and for all its communicative benefits, excessive SNS use has been associated with a range of negative health implications. In the present study, the authors use eye-tracking methodology to explore the relationship between individual differences in personality, mental well-being, SNS usage, and the focus of Facebook users' visual attention. Participants (n = 69, mean age = 23.09, SD = 7.54) completed questionnaire measures for personality and to examine changes in depression, anxiety, stress, and self-esteem. They then engaged in a Facebook session while their eye movements and fixations were recorded. These fixations were coded as being directed to social and update areas of interest (AOI) of the Facebook interface. An exploratory analysis of personality factors revealed a negative correlation between openness to experience and inspection times for the updates AOI and an unexpected negative relationship between extraversion and inspection times for social AOI. There were correlations between changes in depression score and inspection of updated AOI, with reduced depression scores associated with increased inspection of updates. Finally, self-reported duration of participants' typical Facebook sessions did not correlate with eye-tracking measures but were associated with increased Facebook addiction scores and greater increases in depression scores. These initial findings indicate that there are differences in the outcomes of interacting with Facebook which can vary based on Facebook addiction, personality variables, and the Facebook features that individuals interact with.


Endogenous sex hormones, high density lipoprotein cholesterol, and other lipoprotein fractions in men.
At least some of the large preponderance for coronary heart disease in men has been attributed to differences in lipid and lipoprotein levels; notably, high density lipoprotein cholesterol (HDL-C), a protective factor, is higher in women. The sex differences in lipid levels have been postulated to be related to differences in sex hormones. In a southern California cohort of 391 men aged 30-79 years, HDL-C levels were positively correlated and very low density lipoprotein cholesterol (VLDL-C) levels were inversely correlated with testosterone levels independently of age, body mass index, physical exercise, smoking, and alcohol intake. Mean HDL-C levels were 12% higher and VLDL-C levels were 40% lower in the highest compared with the lowest quartile of testosterone level. Low density lipoprotein cholesterol levels were positively related to estrone, estradiol, and androstenedione levels. It is premature to attribute the sex differential in lipid cardiovascular risk profiles to higher levels of testosterone per se in men, since testosterone levels are favorably associated with cardiovascular risk while estrogen levels have the converse relation in men. The differing effects and interactions of specific endogenous sex hormones in men and women require further elucidation.


Narrow linewidth two-electrode 1560 nm laterally coupled distributed feedback lasers with third-order surface etched gratings.
We report on the design and characterization of a re-growth free InGaAsP/InP multiple quantum well two-electrode laterally coupled distributed feedback (LC-DFB) lasers. Third-order surface etched gratings have been defined on the ridge sidewalls along the laser cavity by means of stepper lithography. The lasers oscillate in single-mode around 1560 nm with high side mode suppression ratios (>52 dB), a wavelength tuning (≥ 3nm), an output power (≥ 6 mW), and narrow linewidth (<170 kHz) under various current injection ranges at room temperature. A minimum linewidth of 94 kHz has been recorded for 1500 µm-long two-electrode LC-DFB laser while providing non-uniform current injection through the two electrodes. The effect of the width of the inter-electrode gap on these different performance measures is also studied.


A Guide to Understanding Reimbursement and Value-Based Care in the Military Health System.
With the continued rise in the cost of U.S. health care, there is an increased emphasis on value-based care methodologies. Value is defined as health outcomes achieved per dollar spent. Few studies have evaluated the role of value-based care in the Military Health System (MHS), especially in a format which physicians and providers can understand. The purpose of this article is to provide a guide to understanding current reimbursement systems and value-based care in the MHS and discuss potential strategies for improving value and military readiness. We outlined the current value-based care methodologies in the MHS, and by using musculoskeletal care as an example, offer strategies for further improvement. The MHS has been a leader in the health care industry in adopting value-based care strategies. Current value-based systems in the MHS are primarily designed to incentivize process measure compliance. Initial steps toward measurement and reporting health outcomes have been made, however, with the military's use of the Integrated Resourcing and Incentive System (IRIS), National Surgical Quality Improvement Program (NSQIP) database, and the Joint Outpatient Experience Survey (JOES). As this article will describe, universal reporting of health outcomes, adoption of integrated practice units, and a focus on determining outcomes of illness over the entire care cycle offer a significant opportunity to accelerate the MHS journey to providing true value-based care. The universal measurement and systematic improvement of outcomes based on this measurement will contribute to military medical readiness and warfighter effectiveness.


Systematic analysis of somatic mutations impacting gene expression in 12 tumour types.
We present a novel hierarchical Bayes statistical model, xseq, to systematically quantify the impact of somatic mutations on expression profiles. We establish the theoretical framework and robust inference characteristics of the method using computational benchmarking. We then use xseq to analyse thousands of tumour data sets available through The Cancer Genome Atlas, to systematically quantify somatic mutations impacting expression profiles. We identify 30 novel cis-effect tumour suppressor gene candidates, enriched in loss-of-function mutations and biallelic inactivation. Analysis of trans-effects of mutations and copy number alterations with xseq identifies mutations in 150 genes impacting expression networks, with 89 novel predictions. We reveal two important novel characteristics of mutation impact on expression: (1) patients harbouring known driver mutations exhibit different downstream gene expression consequences; (2) expression patterns for some mutations are stable across tumour types. These results have critical implications for identification and interpretation of mutations with consequent impact on transcription in cancer.


Acute and chronic pituitary failure resembling Sheehan's syndrome following bites by Russell's viper in Burma.
Pituitary function was investigated in 9 patients in shock after Russell's viper bites and in 24 individuals who had been severely envenomed 2 weeks to 24 years previously. 3 out of 9 patients had hypoglycaemia and inappropriately low serum cortisol, plasma growth hormone, and plasma prolactin concentrations. 4 who died had pituitary haemorrhage and 1 had adrenal haemorrhage as well. Of the 24 who had apparently recovered from bites, 7 had clinical features of hypopituitarism and no response in plasma growth hormone or prolactin concentrations to symptom-producing insulin-induced hypoglycaemia. 4 of these 7 had a sluggish serum cortisol response to 'Synacthen Depot' and 5 had an abnormal cortisol response to hypoglycaemia. 4 men with symptoms who were tested had low serum testosterone concentrations; serum thyroxine was also low in these men but not in 2 women with menstrual disturbances and impaired insulin responses. Of the 17 individuals without clinical evidence of endocrine disease, 4 had pituitary hormonal abnormalities. Russell's viper envenoming may thus produce a disorder resembling Sheehan's syndrome.


The role of supplemental coverage in a universal health insurance system: Some Canadian evidence.
To examine the effect of supplemental health insurance for prescription drug coverage on health care utilization as measured by the number of visits to physicians in a setting with incomplete public insurance coverage. A latent-class modeling approach is used to capture the presence of latent heterogeneity in the utilization of physician services. The insurance variable is grouped into three different types, depending upon how it is provided - by government, employers, or private companies. The data for this study come from the Ontario component of the Canadian Community Health Survey 2005, a representative sample of the Ontario population, conducted by Statistics Canada. We find that physician health care utilization responds to the presence and type of insurance, and that the results vary substantially across different types of individuals based on unobservable health status characterized by two latent classes: low users (healthy) and high users (less healthy). The fact that not all individuals have access to supplemental insurance for prescription drug coverage calls into question the universality of public insurance that does not cover important complementary services, such as outpatient prescription drugs.


PDZ domain-mediated interactions of G protein-coupled receptors with postsynaptic density protein 95: quantitative characterization of interactions.
G protein-coupled receptors (GPCRs) constitute the largest family of membrane proteins in the human genome. Their signaling is regulated by scaffold proteins containing PDZ domains, but although these interactions are important for GPCR function, they are still poorly understood. We here present a quantitative characterization of the kinetics and affinity of interactions between GPCRs and one of the best characterized PDZ scaffold proteins, postsynaptic density protein 95 (PSD-95), using fluorescence polarization (FP) and surface plasmon resonance (SPR). By comparing these in vitro findings with colocalization of the full-length proteins in cells and with previous studies, we suggest that the range of relevant interactions might extend to interactions with K i = 450 µM in the in vitro assays. Within this range, we identify novel PSD-95 interactions with the chemokine receptor CXCR2, the neuropeptide Y receptor Y2, and four of the somatostatin receptors (SSTRs). The interaction with SSTR1 was further investigated in mouse hippocampal neurons, where we found a clear colocalization between the endogenously expressed proteins, indicating a potential for further investigation of the role of this interaction. The approach can easily be transferred to other receptors and scaffold proteins and this could help accelerate the discovery and quantitative characterization of GPCR-PDZ interactions.


Clearance Kinetics and Clearance Routes of Molecules From the Suprachoroidal Space After Microneedle Injection.
To determine clearance kinetics and routes of clearance of molecules from the suprachoroidal space (SCS) of live New Zealand White rabbits. Suprachoroidal space collapse rate and pressure changes after microneedle injection into SCS were determined. Fluorescent fundus images were acquired to determine clearance rates of molecules ranging in size from 332 Da to 2 MDa. Microneedle injections of fluorescein were performed, and samples were taken from various sites over time to determine amount of fluorescein exiting the eye. Clearance transport was modeled theoretically and compared with experimental data. After injection, pressures in SCS and vitreous humor spiked and returned to baseline within 20 minutes; there was no difference between these two pressures. Suprachoroidal space collapse occurred within 40 minutes. One hour after fluorescein injection, 46% of fluorescein was still present in the eye, 15% had transported across sclera, 6% had been cleared by choroidal vasculature, and 4% had exited via leakage pathways. Characteristic clearance time increased in proportion with molecular radius, but total clearance of 2 MDa FITC-dextran was significantly slower (21 days) than smaller molecules. These data generally agreed with predictions from a theoretical model of molecular transport. Guided by experimental data in the context of model predictions, molecular clearance from SCS occurred in three regimes: (1) on a time scale of approximately 10 minutes, fluid and molecules exited SCS by diffusion into sclera and choroid, and by pressure-driven reflux via transscleral leakage sites; (2) in approximately 1 hour, molecules cleared from choroid by blood flow; and (3) in 1 to 10 hours, molecules cleared from sclera by diffusion and convection.


Urinary Problems Amongst Gynecological Consultations. Association Between Prolapse, Gynecological Surgery and Diabetes.
Urinary incontinence is the inability of a woman to maintain bladder control. Symptoms range from urgency, frequency, nocturia to urge incontinence (1). It limits functional and social activities and leads to depression and social withdrawal. (2). This observational study aimed to describe the common urinary problems amongst gynecological consultations. It also describes the relationship of urinary incontinence with history of diabetes, previous gynecological surgery and prolapse. The study was conducted as a descriptive cross sectional study from Jan-May 2015 at Qassim University Clinic, Buraidah. Women with urinary problem and those without urinary problems were compared for risk factors including diabetes, prolapse and previous gynecological surgery. The Statistical Package for the Social Sciences 22 (SPSS 22) was used to conduct proportion z-tests to determine the association of prolapse, gynecological surgeries and diabetes with urinary incontinence. To test the hypothesis, differences between two groups on the aforementioned factors were examined. The groups included participants that reported having urinary problems (n = 111) and those who do not have urinary problems (n = 100). The most frequent complaints of participants with urinary problems were urgency (n = 66, 59.46%), Stress incontinence (SI) (n = 65, 58.56%) and frequency (n = 62, 55.86%). For participants with a urinary problem, 89.19% have not had a gynecological surgery (n = 99) and the remaining 10.81% of participants had a gynecological surgery (n = 12). For participants without a urinary problem, 97.0% have not had a gynecological surgery (n = 97) and the remaining 3.0% had a gynecological surgery (n = 3). For participants with a urinary problem, 72.97% did not have diabetes (n = 81) and the remaining 27.03% of participants did have diabetes (n = 30). For participants without a urinary problem, 92.0% did not have diabetes (n = 92) and the remaining 8.0% did report having diabetes (n = 8). For participants with a urinary problem, 91.89% had not experienced a prolapse (n = 102) and the remaining 8.11% of participants had experienced a prolapse (n = 9). For participants without a urinary problem, 98.0% have not experienced a prolapse (n = 98) and the remaining two participants had experienced a prolapse. Gynecological conditions such as prolapse of the uterus, surgery involving the pelvic floor as well as medical conditions such as diabetes mellitus are some of the key risk factors for urinary incontinence. Therefore control of these risk factors can avoid development of urinary incontinence. Bladder retraining, Kegel exercises, drinking of less fluids are some of the preventive measures to avoid development of this embarrassing condition.


LEAP2 changes with body mass and food intake in humans and mice.
Acyl-ghrelin administration increases food intake, body weight, and blood glucose. In contrast, mice lacking ghrelin or ghrelin receptors (GHSRs) exhibit life-threatening hypoglycemia during starvation-like conditions, but do not consistently exhibit overt metabolic phenotypes when given ad libitum food access. These results, and findings of ghrelin resistance in obese states, imply nutritional state dependence of ghrelin's metabolic actions. Here, we hypothesized that liver-enriched antimicrobial peptide-2 (LEAP2), a recently characterized endogenous GHSR antagonist, blunts ghrelin action during obese states and postprandially. To test this hypothesis, we determined changes in plasma LEAP2 and acyl-ghrelin due to fasting, eating, obesity, Roux-en-Y gastric bypass (RYGB), vertical sleeve gastrectomy (VSG), oral glucose administration, and type 1 diabetes mellitus (T1DM) using humans and/or mice. Our results suggest that plasma LEAP2 is regulated by metabolic status: its levels increased with body mass and blood glucose and decreased with fasting, RYGB, and in postprandial states following VSG. These changes were mostly opposite of those of acyl-ghrelin. Furthermore, using electrophysiology, we showed that LEAP2 both hyperpolarizes and prevents acyl-ghrelin from activating arcuate NPY neurons. We predict that the plasma LEAP2/acyl-ghrelin molar ratio may be a key determinant modulating acyl-ghrelin activity in response to body mass, feeding status, and blood glucose.


Analyses of Histone Proteoforms Using Front-end Electron Transfer Dissociation-enabled Orbitrap Instruments.
Histones represent a class of proteins ideally suited to analyses by top-down mass spectrometry due to their relatively small size, the high electron transfer dissociation-compatible charge states they exhibit, and the potential to gain valuable information concerning combinatorial post-translational modifications and variants. We recently described new methods in mass spectrometry for the acquisition of high-quality MS/MS spectra of intact proteins (Anderson, L. C., English, A. M., Wang, W., Bai, D. L., Shabanowitz, J., and Hunt, D. F. (2015) Int. J. Mass Spectrom. 377, 617-624). Here, we report an extension of these techniques. Sequential ion/ion reactions carried out in a modified Orbitrap Velos Pro/Elite(TM) capable of multiple fragment ion fills of the C-trap, in combination with data-dependent and targeted HPLC-MS experiments, were used to obtain high resolution MS/MS spectra of histones from butyrate-treated HeLa cells. These spectra were used to identify several unique intact histone proteoforms with up to 81% sequence coverage. We also demonstrate that parallel ion parking during ion/ion proton transfer reactions can be used to separate species of overlapping m/z that are not separated chromatographically, revealing previously indiscernible signals. Finally, we characterized several truncated forms of H2A and H2B found within the histone fractions analyzed, achieving up to 93% sequence coverage by electron transfer dissociation MS/MS. Results of follow-up in vitro experiments suggest that some of the truncated histone H2A proteoforms we observed can be generated by cathepsin L, an enzyme known to also catalyze clipping of histone H3.


Variability and determinants of total homocysteine concentrations in plasma in an elderly population.
The variability of plasma total homocysteine (tHcy) was examined in 96 individuals over a 1-yr period. Blood tHcy concentrations varied from 7.1 micromol/L in the bottom quintile to 14.5 micromol/L in the top quintile. The mean tHcy was 10.4 micromol/L, the between-person SD was 2.5 micromol/L, and the within-person SD was 0.93 micromol/L. There was little seasonal variation, and the reliability coefficient was 0.88. Mean tHcy concentrations were inversely related to mean plasma folate (r = -0.36) and vitamin B12 (r = -0.35) concentrations. Median tHcy concentrations were approximately 1 micromol/L higher in men than in women and in older (70 to 74 years) than in younger (65 to 69 years) individuals and higher in those with the TT and CT genotypes for the methylenetetrahydrofolate reductase polymorphism than in those with the CC genotype (10.7 and 10.6 vs 9.6 micromol/L). Epidemiological studies based on single tHcy measurements may underestimate the magnitude of any risk associations with disease by 10-15%.


Power laws in citation distributions: evidence from Scopus.
Modeling distributions of citations to scientific papers is crucial for understanding how science develops. However, there is a considerable empirical controversy on which statistical model fits the citation distributions best. This paper is concerned with rigorous empirical detection of power-law behaviour in the distribution of citations received by the most highly cited scientific papers. We have used a large, novel data set on citations to scientific papers published between 1998 and 2002 drawn from Scopus. The power-law model is compared with a number of alternative models using a likelihood ratio test. We have found that the power-law hypothesis is rejected for around half of the Scopus fields of science. For these fields of science, the Yule, power-law with exponential cut-off and log-normal distributions seem to fit the data better than the pure power-law model. On the other hand, when the power-law hypothesis is not rejected, it is usually empirically indistinguishable from most of the alternative models. The pure power-law model seems to be the best model only for the most highly cited papers in "Physics and Astronomy". Overall, our results seem to support theories implying that the most highly cited scientific papers follow the Yule, power-law with exponential cut-off or log-normal distribution. Our findings suggest also that power laws in citation distributions, when present, account only for a very small fraction of the published papers (less than 1 % for most of science fields) and that the power-law scaling parameter (exponent) is substantially higher (from around 3.2 to around 4.7) than found in the older literature.


Beyond the black box: drug- and device-associated hypersensitivity events.
Drug- and device-associated hypersensitivity reactions are serious toxicities that can result in respiratory failure or acute cardiac ischemic events, or even severe hypersensitivity syndromes such as Stevens-Johnson syndrome. These toxicities are usually poorly described in the "black box" warnings section of the product labels. Adverse event reports contained in databases maintained by the Project on Medical Research on Adverse Drug Events and Reports (Med-RADAR), product labels, safety advisories disseminated by pharmaceutical manufacturers, the Food and Drug Administration (FDA), and the Centers for Disease Control and Prevention (CDC) were reviewed. Adverse event reports identified three health care workers who developed nevirapine-associated Stevens-Johnson syndrome following occupational exposure to HIV-infected blood or blood products; four persons with localized hypersensitivity and fatal cardiac events associated with rapamycin- or paclitaxel-coated coronary artery stent placements; and six persons with breast cancer who developed severe or fatal anaphylaxis after receiving adjuvant chemotherapy with Cremophor-EL containing paclitaxel. Safety advisories from the FDA, CDC, and the relevant pharmaceutical manufacturers were ambiguous in their description in "black box" warning sections of package inserts describing these serious and potentially fatal toxicities. Improvements are needed in pharmacovigilance and subsequent dissemination of safety advisories for drug/device-associated hypersensitivity reactions.


Supervised classification of bradykinesia in Parkinson's disease from smartphone videos.
Slowness of movement, known as bradykinesia, is the core clinical sign of Parkinson's and fundamental to its diagnosis. Clinicians commonly assess bradykinesia by making a visual judgement of the patient tapping finger and thumb together repetitively. However, inter-rater agreement of expert assessments has been shown to be only moderate, at best. We propose a low-cost, contactless system using smartphone videos to automatically determine the presence of bradykinesia. We collected 70 videos of finger-tap assessments in a clinical setting (40 Parkinson's hands, 30 control hands). Two clinical experts in Parkinson's, blinded to the diagnosis, evaluated the videos to give a grade of bradykinesia severity between 0 and 4 using the Unified Pakinson's Disease Rating Scale (UPDRS). We developed a computer vision approach that identifies regions related to hand motion and extracts clinically-relevant features. Dimensionality reduction was undertaken using principal component analysis before input to classification models (Naïve Bayes, Logistic Regression, Support Vector Machine) to predict no/slight bradykinesia (UPDRS = 0-1) or mild/moderate/severe bradykinesia (UPDRS = 2-4), and presence or absence of Parkinson's diagnosis. A Support Vector Machine with radial basis function kernels predicted presence of mild/moderate/severe bradykinesia with an estimated test accuracy of 0.8. A Naïve Bayes model predicted the presence of Parkinson's disease with estimated test accuracy 0.67. The method described here presents an approach for predicting bradykinesia from videos of finger-tapping tests. The method is robust to lighting conditions and camera positioning. On a set of pilot data, accuracy of bradykinesia prediction is comparable to that recorded by blinded human experts.


Anthropogenic noise is associated with reductions in the productivity of breeding Eastern Bluebirds (Sialia sialis).
Although previous studies have related variations in environmental noise levels with alterations in communication behaviors of birds, little work has investigated the potential long-term implications of living or breeding in noisy habitats. However, noise has the potential to reduce fitness, both directly (because it is a physiological stressor) and indirectly (by masking important vocalizations and/or leading to behavioral changes). Here, we quantified acoustic conditions in active breeding territories of male Eastern Bluebirds (Sialia sialis). Simultaneously, we measured four fitness indicators: cuckoldry rates, brood growth rate and condition, and number of fledglings produced (i.e., productivity). Increases in environmental noise tended to be associated with smaller brood sizes and were more strongly related to reductions in productivity. Although the mechanism responsible for these patterns is not yet clear, the breeding depression experienced by this otherwise disturbance-tolerant species indicates that anthropogenic noise may have damaging effects on individual fitness and, by extraction, the persistence of populations in noisy habitats. We suggest that managers might protect avian residents from potentially harmful noise by keeping acoustically dominant anthropogenic habitat features as far as possible from favored songbird breeding habitats, limiting noisy human activities, and/or altering habitat structure in order to minimize the propagation of noise pollution.


The rat albumin promoter: cooperation with upstream elements is required when binding of APF/HNF1 to the proximal element is partially impaired by mutation or bacterial methylation.
We have characterized in the accompanying paper (P. Herbomel, A. Rollier, F. Tronche, M.-O. Ott, M. Yaniv, and M. C. Weiss, Mol. Cell. Biol. 9:4750-4758, 1989) six different elements in the albumin promoter. One of them, the proximal element (PE), is the binding site for a strictly liver specific factor, APF/HNF1. This binding site contains a bacterial DAM DNA methylase methylation target sequence which, when methylated, decreases the affinity of the protein for this element. When the different albumin promoter constructions were prepared in an Escherichia coli deoxyadenosine methylase-negative strain, the respective contributions of the elements to the overall promoter activity were strikingly different. An intact proximal element plus the TATA box gave almost full transcriptional activity in transient transfection experiments and only in differentiated hepatoma cells of line H4II, whereas the distal elements (distal element III [DEIII], the NF1-binding site DEII, and the E/CBP-binding site DEI) had become essentially dispensable. Mutations affecting the CCAAT box showed only a two- to threefold decrease. When PE was methylated, mutated, or replaced by the homologous element from the alpha-fetoprotein gene, activity in the context of the short promoter (PE plus the TATA box) was abolished. However, activity was restored in the presence of the upstream elements, showing that cooperation with factors binding to the CCAAT box and distal elements favors the functional interaction of the liver-specific APF/HNF1 factor with lower-affinity binding sites.


Computational Evidence Suggests That 1-Chloroethanol May Be an Intermediate in the Thermal Decomposition of 2-Chloroethanol into Acetaldehyde and HCl.
The dehalogenation of 2-chloroethanol (2ClEtOH) in the gas phase with and without the participation of catalytic water molecules has been investigated using methods rooted into the density functional theory. The well-known HCl elimination leading to vinyl alcohol (VA) was compared to the alternative elimination route toward oxirane and shown to be kinetically and thermodynamically more favorable. However, the isomerization of VA to acetaldehyde in the gas phase, in the absence of water, was shown to be kinetically and thermodynamically less favorable than the recombination of VA and HCl to form the isomeric 1-chloroethanol (1ClEtOH) species. At the ωB97X-D/cc-pVTZ level of calculation, this species is more stable than 2ClEtOH by about 6 kcal mol-1 at 298 K, and the reaction barrier for VA to 1ClEtOH is 23 kcal mol-1 versus 55 kcal mol-1 for the direct transformation of VA to acetaldehyde. In a successive step, 1ClEtOH can decompose directly to acetaldehyde and HCl with a lower barrier (29 kcal mol-1) than that of VA to the same products (55 kcal mol-1). The calculations were repeated using a single ancillary water molecule (W) in the complexes 2ClEtOH_W and 1ClEtOH_W. The latter adduct is now more stable than 2ClEtOH_W by about 8 kcal mol-1 at 298 K, implying that the water molecule increased the already higher stability of 1ClEtOH in the gas phase. However, this catalytic water molecule lowers dramatically the barrier for the interconversion of VA to acetaldehyde (from 55 to 7 kcal mol-1). This barrier is now smaller than the one for the conversion to 1ClEtOH (which also decreases, but not so much, from 23 to 13 kcal mol-1). Thus, it is concluded that while 1ClEtOH may be a plausible intermediate in the gas phase dehalogenation of 2ClEtOH, it is unlikely that it plays a major role in water complexes (or, by inference, aqueous solution). It is also shown that neither in the gas phase nor in the cluster with one water molecule, the oxirane path is more favorable than the VA alcohol path. Additionally, a direct conversion of 2ClEtOH to 1ClEtOH through a transition state which resembles a VA molecule in a complex with a chlorine atom and a hydrogen atom on both sides of this planar species was found. This reaction path has also lower activation energy than the conversion to oxirane but not as low as the conversion to VA.


Retail supply of malaria-related drugs in rural Tanzania: risks and opportunities.
To characterize availability of fever and malaria medicines within the retail sector in rural Tanzania, assess the likely public health implications, and identify opportunities for policy interventions to increase the coverage of effective treatment. A census of retailers selling drugs was undertaken in the areas under demographic surveillance in four Tanzanian districts, using a structured questionnaire. Drugs were stocked by two types of retailer: a large number of general retailers (n = 675) and a relatively small number of drug shops (n = 43). Almost all outlets stocked antipyretics/painkillers. One-third of general retailers stocking drugs had antimalarials, usually chloroquine alone. Almost all drug shops stocked antimalarials (98%): nearly all had chloroquine, 42% stocked quinine, 37% sulphadoxine-pyrimethamine and 30% amodiaquine. A large number of antimalarial brands were available. Population ratios indicate the relative accessibility of retail drug providers compared with health facilities. Drug shop staff generally travelled long distances to buy from drugs wholesalers or pharmacies. General retailers bought mainly from local general wholesalers, with a few general wholesalers accounting for a high proportion of all sources cited. Drugs were widely available from a large number of retail outlets. Potential negative implications include provision of ineffective drugs, confusion over brand names, uncontrolled use of antimalarials, and the availability of components of potential combination therapy regimens as monotherapies. On the other hand, this active and highly accessible retail market provides opportunities for improving the coverage of effective antimalarial treatment. Interventions targeted at all drug retailers are likely to be costly to deliver and difficult to sustain, but two promising points for targeted intervention are drug shops and selected general wholesalers. Retail quality may also be improved through consumer education, and modification of the chemical quality, packaging and price of products entering the retail distribution chain.


Thyroid hormone targets matrix Gla protein gene associated with vascular smooth muscle calcification.
Thyroid hormones have marked cardiovascular effects in vivo. However, their direct effects on vascular smooth muscle cells have been unclear. Because thyroid hormones play critical roles in bone remodeling, we hypothesized that they are also associated with vascular smooth muscle calcification, one of the pathological features of vascular sclerosis. To test this hypothesis, we examined the effects of 3',3,5-triiodo-L-thyronine (T3) on the expression of calcification-associated genes in rat aortic smooth muscle cells (RAOSMCs). Quantitative RT-PCRs revealed that a physiological concentration of T3 (15 pmol/L free T3) increased mRNA level of matrix Gla protein (MGP), which acts as a potent inhibitor of vascular calcification in vivo, by 3-fold in RAOSMCs, as well as in cultured human coronary artery smooth muscle cells. In RAOSMCs transiently transfected with a luciferase reporter gene driven by the MGP promoter, T3 significantly stimulated luciferase activity. In addition, RNA interference against thyroid hormone receptor-alpha gene diminished the effect of T3 on MGP expression. Aortic smooth muscle tissues from methimazole-induced hypothyroid rats (400 mg/L drinking water; 4 weeks) also showed a 68% decrease in the MGP mRNA level, as well as a 33% increase in calcium content compared with that from the control euthyroid animals, whereas hyperthyroidism (0.2 mg T3/kg IP; 10 days) upregulated MGP mRNA by 4.5-fold and reduced calcium content by 11%. Our findings suggest that a physiological concentration of thyroid hormone directly facilitates MGP gene expression in smooth muscle cells via thyroid hormone nuclear receptors, leading to prevention of vascular calcification in vivo.


Cyclic peptides target the aromatic cage of a PHD-finger reader domain to modulate epigenetic protein function.
Plant homeodomain fingers (PHD-fingers) are a family of reader domains that can recruit epigenetic proteins to specific histone modification sites. Many PHD-fingers recognise methylated lysines on histone tails and play crucial roles in transcriptional regulation, with their dysregulation linked to various human diseases. Despite their biological importance, chemical inhibitors for targeting PHD-fingers are very limited. Here we report a potent and selective de novo cyclic peptide inhibitor (OC9) targeting the Nε-trimethyllysine-binding PHD-fingers of the KDM7 histone demethylases, developed using mRNA display. OC9 disrupts PHD-finger interaction with histone H3K4me3 by engaging the Nε-methyllysine-binding aromatic cage through a valine, revealing a new non-lysine recognition motif for the PHD-fingers that does not require cation-π interaction. PHD-finger inhibition by OC9 impacted JmjC-domain mediated demethylase activity at H3K9me2, leading to inhibition of KDM7B (PHF8) but stimulation of KDM7A (KIAA1718), representing a new approach for selective allosteric modulation of demethylase activity. Chemoproteomic analysis showed selective engagement of OC9 with KDM7s in T cell lymphoblastic lymphoma SUP T1 cells. Our results highlight the utility of mRNA-display derived cyclic peptides for targeting challenging epigenetic reader proteins to probe their biology, and the broader potential of this approach for targeting protein-protein interactions.


Iodine toxicity treated with hemodialysis and continuous venovenous hemodiafiltration.
Continuous mediastinal irrigation with povidone-iodine is used commonly for treating severe postoperative mediastinitis. However, concurrent iodine toxicity has been reported, particularly in patients with renal dysfunction (likely because absorbed iodine is renally excreted). The authors were consulted on a 45-year-old patient with mediastinitis who had renal and hepatic dysfunction while being treated with mediastinal irrigation of povidone-iodine. The povidone-iodine irrigation was discontinued because he had toxic plasma iodine levels. Despite this, his condition worsened, and the iodine levels remained elevated. Thus, hemodialysis (HD) was initiated using high-flux membranes followed by continuous venovenous hemodiafiltration (CVVHDF; 2 L/h of hemofiltration and 2 L/h of HD). Plasma and effluent iodine levels were measured repeatedly to determine iodine clearance by these 2 modalities (HD, 120 mL/min; CVVHDF, 37 mL and 44 mL/min on days 1 and 2, respectively). Hepatic and renal functions improved with decreasing plasma iodine levels. Based on this experience and after reviewing the literature the authors conclude that: (1) iodine irrigation can increase blood iodine levels significantly, especially in the setting of renal failure, and lead to increased morbidity and mortality; (2) plasma iodine levels should be monitored in patients with renal insufficiency; and (3) HD and CVVHDF are effective at clearing iodine. The authors suggest that patients that are at high risk or already developing signs of iodine toxicity should have the iodine irrigation discontinued and may benefit from renal replacement therapy (RRT). Alternatively, concomitant RRT during iodine irrigation may be attempted to maintain the systemic iodine levels at nontoxic levels.


Palliative care for patients with locally advanced and metastatic non-small cell lung cancer.
Non-small cell lung cancer (NSCLC) often presents at an advanced stage and can result in a significant symptomatic burden. Many patients with advanced NSCLC experience symptoms that include pain, dyspnea, cough, decreased appetite, weight loss, and depression. Early initiation of palliative care for advanced or metastatic NSCLC can reduce symptoms, improve quality of life, and prolong survival. Palliative care interventions by an interdisciplinary team focus on patient and family and address physical symptoms, illness understanding, coping, and psychosocial and spiritual distress. The addition of systemic therapy to best supportive care can further improve quality of life and prolong survival. Thoracic external beam radiation therapy beam radiotherapy is well tolerated and can improve quality of life and relieve symptoms such as hemoptysis, pain, cough, and dyspnea. For central airway obstruction, external beam radiation therapy, endobronchial brachytherapy, Nd:YAG laser therapy, and photodynamic therapy can improve symptoms. Symptomatic lung metastases to the brain, adrenal glands, and liver or compressing or impinging the spinal cord or nerve roots can also result in patient symptoms and can be managed with medical and pharmacologic interventions, radiation therapy, surgery, ablative therapy, and systemic therapy. All healthcare providers treating patients with advanced or metastatic NSCLC should make palliative care a priority by assessing for pain, dyspnea, coping and psychosocial distress, and other symptoms at each patient encounter to reduce patient morbidity, improve quality of life, and potentially prolong survival.


Ephedrine-induced thermogenesis in man: no role for interscapular brown adipose tissue.
The warmest interscapular skin areas were located by thermography in six healthy subjects during ephedrine-induced thermogenesis. In these interscapular areas, and in lumbar control areas, the skin temperature, subcutaneous temperature and adipose tissue blood flow were measured before and during ephedrine-induced thermogenesis. The skin and subcutaneous temperatures increased in the interscapular area as well as in the lumbar area, by about 0.7-1.2 degrees C. The interscapular skin temperature remained about 1 degree C higher than the lumbar; the subcutaneous temperatures in the two areas were identical during the experiments. Although the interscapular subcutaneous adipose tissue blood flow increased about sixfold and the lumbar increased twofold, the absolute flows were higher in the lumbar area. The oxygen uptake increased to a maximum of 30% above control level. Plasma glucose and glycerol concentrations remained unchanged, and plasma non-esterified fatty acids, lactate and noradrenaline concentrations increased slightly but significantly. Biopsies taken from the hot interscapular areas did not contain brown adipose tissue. It is concluded that the high interscapular skin temperature may be due to a lower insulating fat thickness and that the increases in skin and subcutaneous temperatures during ephedrine-induced thermogenesis are caused by an increased blood flow. These observations weigh against the hypothesis that the interscapular temperature increase is due to functional, interscapular brown adipose tissue.


Gastrointestinal tolerance and utilization of agave inulin by healthy adults.
Little clinical research exists on agave inulin as a fiber source. Due to differences in botanical origin and chemical structure compared to other inulin-type fibers, research is needed to assess gastrointestinal (GI) tolerance following consumption. This study aimed to evaluate GI tolerance and utilization of 5.0 and 7.5 g per day of agave inulin in healthy adults (n = 29) using a randomized, double-blind, placebo-controlled crossover trial consisting of three 21 day periods with 1 week washouts among periods. GI tolerance was assessed via daily and weekly questionnaires, three fecal samples were collected on days 16-20 of each period, and breath hydrogen testing was completed on the final day of each treatment period. Survey data were compared using a generalized linear mixed model. All other outcomes were analyzed using a mixed linear model with a repeated measures procedure. Composite GI intolerance scores for 5.0 and 7.5 g treatments were both greater (P < 0.05) than control, however, scores were low, with means of 0.4, 1.9, and 2.3 on a 0-12 point composite scale for 0, 5.0, and 7.5 g treatments, respectively. There were slight increases (P < 0.05) in bloating, flatulence, and rumbling frequency with 5.0 and 7.5 g agave inulin. Abdominal pain and rumbling intensity were marginally greater (P < 0.05) with 7.5 g. Bloating and flatulence intensity increased (P < 0.05) with 5.0 g and 7.5 g. Agave inulin did not affect diarrhea (P > 0.05). Number of bowel movements per day increased, stools were softer, and stool dry matter percentage was lower with 7.5 g (P < 0.05). Breath hydrogen concentrations increased (P < 0.001) from 5-8 hour postprandial when participants consumed agave inulin compared to control. These data demonstrate that doses up to 7.5 g per day of agave inulin led to minimal GI upset, do not increase diarrhea, and improve laxation in healthy young adults.


Deficiency in cold-inducible RNA-binding protein attenuates acute respiratory distress syndrome induced by intestinal ischemia-reperfusion.
Intestinal ischemia-reperfusion can occur in shock and mesenteric occlusive diseases, causing significant morbidity and mortality. Aside from local injury, intestinal ischemia-reperfusion can result in remote organ damage, particularly in the lungs. Cold-inducible RNA-binding protein (CIRP) was identified as a novel inflammatory mediator. We hypothesized that a deficiency in CIRP would protect the lungs during intestinal ischemia-reperfusion injury. Intestinal ischemia was induced in adult male C57BL/6 wild-type and CIRP knock-out (CIRP-/-) mice via clamping of the superior mesenteric artery for 60 minutes. Reperfusion was allowed for 4 hours or 20 hours, and blood, gut, and lung tissues were harvested for various analyses. After intestinal ischemia-reperfusion, the elevated levels of serum lactate dehydrogenase and inflammatory cytokine interleukin-6 were reduced by 68% and 98%, respectively, at 20 hours after ischemia-reperfusion in CIRP-/- mice compared with the wild-type mice. In the gut, mRNA levels of inflammatory cytokine interleukin-6 were reduced by 67% at 4 hours after ischemia-reperfusion in CIRP-/- mice. In the lungs, inflammatory cytokine interleukin-6 protein and myeloperoxidase activity were reduced by 78% and 26% at 20 hours and 4 hours after ischemia-reperfusion, respectively, in CIRP-/- mice. Finally, the elevated lung caspase-3 was significantly decreased by 55%, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling-positive cells decreased by 91%, and lung injury score decreased by 37% in CIRP-/- mice at 20 hours after ischemia-reperfusion. Increased levels of proinflammatory cytokines, myeloperoxidase, and apoptosis are the hallmarks of acute respiratory distress syndrome. We noticed after intestinal ischemia-reperfusion the proinflammatory milieu in lungs was elevated significantly, while the CIRP-/- mice had significantly decreased levels of proinflammatory cytokine, myeloperoxidase, and apoptotic cells leading to decreased lung injury. These findings strongly established a causal link between CIRP and acute respiratory distress syndrome during intestinal ischemia-reperfusion injuries. Targeting CIRP may therefore be beneficial for treatment of intestinal ischemia-reperfusion-associated acute respiratory distress syndrome acute respiratory distress syndrome.


'We call it a virus but I want to say it's the devil inside': redemption, moral reform and relationships with God among people living with HIV in Papua New Guinea.
There is growing recognition of the importance of religion and religious beliefs as they relate to the experience of HIV, globally and in Papua New Guinea in particular. Based on 36 in-depth qualitative interviews conducted with people living with HIV receiving HIV antiretroviral therapy in 2008, this paper examines the cultural aetiology of HIV of in Papua New Guinea, the country with the highest reported burden of HIV in the Pacific. Narratives provided drew upon a largely moral framework, which viewed HIV acquisition as a consequence of moral failing and living an un-Christian life. This explanation for suffering viewed the individual as responsible for their condition in much the same way that neo-liberal biomedical discourses do. Moral reform and re-establishing a relationship with God were seen as key actions necessary to effect healing on the material body infected with HIV. Religious understandings of HIV drew upon a pre-existing cultural aetiology of dis-ease and misfortune widespread in Papua New Guinea. Understanding the centrality of Christianity to explanations of disease, and subsequently the actions necessary to bring about health, is essential in order to understand how people with HIV in receipt of antiretroviral therapies internalise biomedical perspectives and reconcile these with Christian beliefs.


AN ITALIAN SURVEY OF COMPLIANCE WITH MAJOR GUIDELINES FOR L-THYROXINE OF PRIMARY HYPOTHYROIDISM.
The adherence by endocrinologists to guideline regarding levothyroxine (LT4) therapy and the compliance of patients may impact the management of hypothyroidism. The aim of this study was to compare the adherence of Italian endocrinologists to the ATA/AACE and ETA guidelines on the management of newly diagnosed primary hypothyroidism and to validate the Italian version of the Morisky-Green Medical Adherence Scale-8 (MMAS-8) questionnaire as applied to the evaluation of the adherence of patients with hypothyroidism to LT4 treatment. This was an observational, longitudinal, multicenter, cohort study, involving 12 Italian Units of Endocrinology. The study enrolled 1,039 consecutive outpatients (mean age 48 years; 855 women, 184 men). The concordance of Italian endocrinologists with American Association of Clinical Endocrinologists/American Thyroid Association (AACE/ATA) and European Thyroid Association (ETA) recommendations was comparable (77.1% and 71.7%) and increased (86.7 and 88.6%) after the recommendations on LT4 dose were excluded, considering only the remaining recommendations on diagnosis, therapy, and follow-up. The MMAS-8 was filled out by 293 patients. The mean score was 6.71 with 23.9% low (score <6), 38.6% medium (6 to <8), 37.5% highly (= 8) adherers; the internal validation coefficient was 0.613. Highly adherent patients were not more likely to have good control of hypothyroidism compared with either medium (69% versus 72%, P = .878) or low (69% versus 43%, P = .861) adherers. Clinical management of hypothyroidism in Italy demonstrated an observance of international guidelines by Italian endocrinologists. Validation of the Italian version of the MMAS-8 questionnaire provides clinicians with a reliable and simple tool for assessing the adherence of patients to LT4 treatment. AACE = American Association of Clinical Endocrinologists; ATA = American Thyroid Association; EDIPO = Endotrial SIE: DIagnosis and clinical management of Primitive hypothyrOidism in Italy; eCRF = electronic case report form; ETA = European Thyroid Association; fT3 = free triiodothyronine; fT4 = free thyroxine; LT4 = levothyroxine; MMAS-8 = Morisky-Green Medical Adherence Scale-8; PH = primary hypothyroidism; T3 = triiodothyronine; T4 = thyroxine; TSH = thyroid-stimulating hormone; US = ultrasonography.


Efficient adaptation of complex-valued noiselet sensing matrices for compressed single-pixel imaging.
Minimal mutual coherence of discrete noiselets and Haar wavelets makes this pair of bases an essential choice for the measurement and compression matrices in compressed-sensing-based single-pixel detectors. In this paper, we propose an efficient way of using complex-valued and nonbinary noiselet functions for object sampling in single-pixel cameras with binary spatial light modulators and incoherent illumination. The proposed method allows us to determine m complex noiselet coefficients from m+1 binary sampling measurements. Further, we introduce a modification to the complex fast noiselet transform, which enables computationally efficient real-time generation of the binary noiselet-based patterns using efficient integer calculations on bundled patterns. The proposed method is verified experimentally with a single-pixel camera system using a binary spatial light modulator.


Caffeine protects Alzheimer's mice against cognitive impairment and reduces brain beta-amyloid production.
A recent epidemiological study suggested that higher caffeine intake over decades reduces the risk of Alzheimer's disease (AD). The present study sought to determine any long-term protective effects of dietary caffeine intake in a controlled longitudinal study involving AD transgenic mice. Caffeine (an adenosine receptor antagonist) was added to the drinking water of amyloid precursor protein, Swedish mutation (APPsw) transgenic (Tg) mice between 4 and 9 months of age, with behavioral testing done during the final 6 weeks of treatment. The average daily intake of caffeine per mouse (1.5 mg) was the human equivalent of 500 mg caffeine, the amount typically found in five cups of coffee per day. Across multiple cognitive tasks of spatial learning/reference memory, working memory, and recognition/identification, Tg mice given caffeine performed significantly better than Tg control mice and similar to non-transgenic controls. In both behaviorally-tested and aged Tg mice, long-term caffeine administration resulted in lower hippocampal beta-amyloid (Abeta) levels. Expression of both Presenilin 1 (PS1) and beta-secretase (BACE) was reduced in caffeine-treated Tg mice, indicating decreased Abeta production as a likely mechanism of caffeine's cognitive protection. The ability of caffeine to reduce Abeta production was confirmed in SweAPP N2a neuronal cultures, wherein concentration-dependent decreases in both Abeta1-40 and Abeta1-42 were observed. Although adenosine A(1) or A(2A) receptor densities in cortex or hippocampus were not affected by caffeine treatment, brain adenosine levels in Tg mice were restored back to normal by dietary caffeine and could be involved in the cognitive protection provided by caffeine. Our data demonstrate that moderate daily intake of caffeine may delay or reduce the risk of AD.


A quantitative, real-time polymerase chain reaction assay for beak and feather disease virus.
PCR-based assays for the detection of BFDV DNA are in widespread use throughout the world. Quantitative real-time PCR assays are extremely sensitive and have the advantages over standard PCR assays that they do not require post-reaction processing to visualise PCR products and can quantify the amount of DNA present in a sample. This study describes a quantitative real-time PCR assay for the detection of BFDV DNA, using primers designed to amplify a conserved 81 bp fragment of ORFV1 and SYTO9, a fluorescent intercalating dye. A synthetic oligonucleotide was used to make standard curves for the quantitation of viral load in blood and feather preparations. The assay was very sensitive, with a detection limit of 50 copies/microL. The assay was developed using DNA extracts from the feathers of 10 different species of birds which had tested BFDV-positive previously and was validated with blood and feather samples from corellas vaccinated with an experimental BFDV vaccine, then challenged with live virus. Viral DNA was detected consistently in the blood of all control (non-vaccinated) birds and in some vaccinated birds. Contamination of the environment with feather dander from BFDV-infected birds meant that feather preparations used for the haemagglutination assay were unreliable for the detection and quantitation of viral excretion. Nonetheless, the assay should prove to be a useful and sensitive test for the detection of viral DNA in a range of samples.


Smoking and socio-demographic determinant of cardiovascular diseases among males 45+ years in Indonesia.
The main objective of this study is to present the prevalence of Cardio Vascular Diseases (CVDs) defined as been diagnosed or having symptoms of Coronary Heart Disease, Arrhytmia, or Heart Failure. The main risk factor analyzed is smoking behavior. The data used for this study was from Basic Health Survey of 2007, a National baseline data collected every three years which consist of more than one million samples representing 33 provinces in Indonesia. Information on socio-demographic characteristics, history of CVDs and smoking behavior were collected by highly-trained interviewers using a questionnaire which had been tested. A sub-sample of the survey consisting of 100,009 males aged 45 years and over was analyzed. Crude and adjusted odds ratio (OR) were analyzed using logistic regressions to estimate the prevalence of CVDs by smoking behavior and socio-demographic characteristics. Overall, 86.8% respondents reported that they had never been diagnosed as having CVDs or having any symptom of CVDs.; while 2.1% respondents reported that they had been diagnosed by a health professional (a doctor or a nurse) of having CVDs. The interviewers also identified three signs and symptoms of CVDs for all respondents if they reported of never been diagnosed CVDs. Among all respondents 2.3% had symptoms of coronary heart disease, 4.9% had symptoms of arrhytmia, and 3.9% had symptoms of heart failure. The prevalence of CVDs was significantly higher in former smokers (OR = 2.03), and duration of smoking for more than 20 years. The prevalence of CVDs was significantly higher among older groups. Old males who lived in Sulawesi island had higher probability of having CVDs (OR = 1.67). The lower prevalence of CVDs seemed to have associated with higher among Senior High School Graduate compared to those who Never Schooling (OR = 0.8). Since population of Indonesia is relatively young, the future of health care costs of Indonesia would be high due to high prevalence of smoking among males population. This finding suggests that Indonesia should ratify Framework Convention on Tobacco Control ans start impelementing measures to control tobacco uses in order to reduce public health and economic consequences of smoking in the future.


Young children's probability of dying before and after their mother's death: a rural South African population-based surveillance study.
There is evidence that a young child's risk of dying increases following the mother's death, but little is known about the risk when the mother becomes very ill prior to her death. We hypothesized that children would be more likely to die during the period several months before their mother's death, as well as for several months after her death. Therefore we investigated the relationship between young children's likelihood of dying and the timing of their mother's death and, in particular, the existence of a critical period of increased risk. Data from a health and socio-demographic surveillance system in rural South Africa were collected on children 0-5 y of age from 1 January 1994 to 31 December 2008. Discrete time survival analysis was used to estimate children's probability of dying before and after their mother's death, accounting for moderators. 1,244 children (3% of sample) died from 1994 to 2008. The probability of child death began to rise 6-11 mo prior to the mother's death and increased markedly during the 2 mo immediately before the month of her death (odds ratio [OR] 7.1 [95% CI 3.9-12.7]), in the month of her death (OR 12.6 [6.2-25.3]), and during the 2 mo following her death (OR 7.0 [3.2-15.6]). This increase in the probability of dying was more pronounced for children whose mothers died of AIDS or tuberculosis compared to other causes of death, but the pattern remained for causes unrelated to AIDS/tuberculosis. Infants aged 0-6 mo at the time of their mother's death were nine times more likely to die than children aged 2-5 y. The limitations of the study included the lack of knowledge about precisely when a very ill mother will die, a lack of information about child nutrition and care, and the diagnosis of AIDS deaths by verbal autopsy rather than serostatus. Young children in lower income settings are more likely to die not only after their mother's death but also in the months before, when she is seriously ill. Interventions are urgently needed to support families both when the mother becomes very ill and after her death. Please see later in the article for the Editors' Summary.


Promoter upstream elements of the chicken cardiac myosin light-chain 2-A gene interact with trans-acting regulatory factors for muscle-specific transcription.
A segment of the 5'-flanking region of the chicken cardiac myosin light-chain gene extending from nucleotide -64 to the RNA start site is sufficient to allow muscle-specific transcription. In this paper, we characterize, by mutational analysis, sequence elements which are essential for the promoter activity. Furthermore, we present evidence for a negative-acting element which is possibly involved in conferring the muscle specificity. Nuclear proteins specifically bind to the DNA elements, as demonstrated by gel mobility shift assays and DNase I protection footprinting. The significance of the DNA-protein interactions for the function of the promoter in vivo is demonstrated by competition experiments in which protein-binding oligonucleotides were microinjected into nuclei of myotubes, where they successfully competed for the protein factors which are required to trans activate the MLC2-A promoter. The ability to bind nuclear proteins involves two closely spaced AT-rich sequence elements, one of which constitutes the TATA box. The binding properties correlate well with the capacity to activate transcription in vivo, since mutations in this region of the promoter concomitantly lead to loss of binding and transcriptional activity.


Development of porphyria cutanea tarda after treatment with cyclophosphamide.
Porphyria cutanea tarda, a metabolic disorder of heme biosynthesis, is characterized by cutaneous hyperpigmentation, facial hypertrichosis, dark urine, and a distinctive pattern of excess porphyrin production. Hepatic uroporphyrinogen decarboxylase activity is markedly reduced in patients with this disorder. Although porphyria cutanea tarda may be familial, it is more often sporadic in occurrence, and has been associated with excess alcohol ingestion, estrogen administration, iron overload, and several environmental hepatotoxins. It has also been associated on occasion with malignancy. We report a 46-yr-old woman with ovarian carcinoma who developed porphyria cutanea tarda while undergoing treatment with cisplatin and cyclophosphamide. The temporal course of the porphyrin abnormality suggested that cyclophosphamide was the pathogenic agent, and symptoms regressed after cessation of this drug with continued administration of cisplatin. The pathogenesis of the porphyria is not clear; however, cyclophosphamide is a substrate for cytochrome P450, and may produce metabolites that destroy this protein. The resulting increased turnover of heme might then result in overproduction of porphyrin precursors, resulting in the clinical syndrome. Studies of porphyrin metabolism in patients treated with cyclophosphamide may help to elucidate this possibility.


[A patient with vision problems: first manifestation of a neurodegenerative condition].
Vision problems are common and the causes are diverse. This case illustrates the fact that vision problems may also be the first manifestation of a neurodegenerative disorder. A 46-year-old male developed progressive, incapacitating vision problems. Multiple pairs of glasses with lenses of different strengths did not improve symptoms. No ophthalmological explanation of the problems could be found. Oculomotor apraxia and visual extinction were seen on neurological examination. Visuoperceptual impairment was the main finding on neuropsychological examination. Impairment was also identified in visuoconstruction, memory and praxis. Cognitive problems in several areas, interference with activities of daily living, and young age of onset complete the clinical picture of presenile dementia. Posterior cortical atrophy is a spectrum of neurodegenerative disorders, characterized by progressive, incapacitating visuospatial and visuoperceptual impairment. The most prevalent underlying pathology is Alzheimer's disease. At present there is no curative therapy for posterior cortical atrophy.


Thermal rescue of UV-irradiated bacteriophage T4 and biphasic mode of action of the WXY system.
When ultraviolet-irradiated bacteriophage T4 is assayed at plating temperatures ranging from 20 degrees to 40 degrees, its survival increases at the higher temperatures. This "thermal rescue" requires an intact WXY system but not the denV pyrimidine dimer excision system. Mutation rates decrease with increasing temperature, indicating that some lesions processed in a mutagenic manner at lower temperatures are accurately repaired or circumvented at high temperatures. When both the cold sensitivity of UV survival in the wild type and the temperature sensitivity of newly isolated ts mutants of uvsX and uvsY were used, expression of the WXY system was monitored in temperature shift UV survival experiments and was found to be biphasic: the uvsX and uvsY functions increase UV survival in two increments, one at an early and another at a late stage of infection. The uvsW function, however, increases UV survival only early in infection.


The estimation of utility weights in cost-utility analysis for mental disorders: a systematic review.
To systematically review approaches and instruments used to derive utility weights in cost-utility analyses (CUAs) within the field of mental disorders and to identify factors that may have influenced the choice of the approach. We searched the databases DARE (Database of Abstracts of Reviews of Effects), NHS EED (National Health Service Economic Evaluation Database), HTA (Health Technology Assessment), and PubMed for CUAs. Studies were included if they were full economic evaluations and reported quality-adjusted life-years as the health outcome. Study characteristics and instruments used to estimate utility weights were described and a logistic regression analysis was conducted to identify factors associated with the choice of either the direct (e.g. standard gamble) or the preference-based measure (PBM) approach (e.g. EQ-5D). We identified 227 CUAs with a maximum in 2009, 2010, and 2012. Most CUAs were conducted in depression, dementia, or psychosis, and came from the US or the UK, with the EQ-5D being the most frequently used instrument. The application of the direct approach was significantly associated with depression, psychosis, and model-based studies. The PBM approach was more likely to be used in recent studies, dementia, Europe, and empirical studies. Utility weights used in model-based studies were derived from only a small number of studies. We only searched four databases and did not evaluate the quality of the included studies. Direct instruments and PBMs are used to elicit utility weights in CUAs with different frequencies regarding study type, mental disorder, and country.


Blood pressure lowering diminishes age-related changes in the rat aortic intima.
These studies were undertaken to determine whether age-related changes in the aortic intima can be inhibited by prolonged blood pressure lowering. Normotensive Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) were treated with a combination of chlorothiazide, reserpine, and hydralazine beginning at 20 weeks of age and continuing until 62 weeks. Aortic morphology was assessed at the termination of the study by light and electron microscopy. Average systolic blood pressures during the final 6 months of therapy were: for untreated WKY, 147 +/- 5 mm Hg (mean +/- SE); for treated WKY, 110 +/- 2; for untreated SHR, 206 +/- 3; and for treated SHR, 104 +/- 4. Intimal and medial abnormalities were maximal in untreated SHR, while untreated WKY showed moderate changes consistent with their age. In contrast, both treatment groups exhibited relatively minimal alterations despite their advanced age. The occurrence of intimal lesions in individual animals correlated with the average level of blood pressure during therapy. These data suggest that blood pressure reduction, even in normotensive animals, may reduce the effects of aging on the arterial wall.


Mutations Identified in the Hepatitis C Virus (HCV) Polymerase of Patients with Chronic HCV Treated with Ribavirin Cause Resistance and Affect Viral Replication Fidelity.
Ribavirin has been used for 25 years to treat patients with chronic hepatitis C virus (HCV) infection; however, its antiviral mechanism of action remains unclear. Here, we studied virus evolution in a subset of samples from a randomized 24-week trial of ribavirin monotherapy versus placebo in chronic HCV patients, as well as the viral resistance mechanisms of the observed ribavirin-associated mutations in cell culture. Thus, we performed next-generation sequencing of the full-length coding sequences of HCV recovered from patients at weeks 0, 12, 20, 32 and 40 and analyzed novel single nucleotide polymorphisms (SNPs), diversity, and mutation-linkage. At week 20, increased genetic diversity was observed in 5 ribavirin-treated compared to 4 placebo-treated HCV patients due to new synonymous SNPs, particularly G-to-A and C-to-U ribavirin-associated transitions. Moreover, emergence of 14 nonsynonymous SNPs in HCV nonstructural 5B (NS5B) occurred in treated patients, but not in placebo controls. Most substitutions located close to the NS5B polymerase nucleotide entry site. Linkage analysis showed that putative resistance mutations were found in the majority of genomes in ribavirin-treated patients. Identified NS5B mutations from genotype 3a patients were further introduced into the genotype 3a cell-culture-adapted DBN strain for studies in Huh7.5 cells. Specific NS5B substitutions, including DBN-D148N+I363V, DBN-A150V+I363V, and DBN-T227S+S183P, conferred resistance to ribavirin in long-term cell culture treatment, possibly by reducing the HCV polymerase error rate. In conclusion, prolonged exposure of HCV to ribavirin in chronic hepatitis C patients induces NS5B resistance mutations leading to increased polymerase fidelity, which could be one mechanism for ribavirin resistance.


Recurrence of dislocation following total hip arthroplasty revision using dual mobility cups was rare in 180 hips followed over 7 years.
Dual mobility (DM) cups of mobile polyethylene were introduced to prevent total hip arthroplasty (THA) dislocation, but no large series with this design to treat recurrent instability have been reported. Our retrospective investigation ascertained the efficiency of DM cups in correction of recurrent dislocation and assessed any adverse effects. One hundred eighty THAs with recurrent instability were revised to DM cups in 180 patients (mean age, 67.4 ± 11.7 years; range, 19 to 92 years). Thirty-one patients (17.2%) underwent at least one earlier THA revision, and 15 (10.3%) incurred non-union of the greater trochanter. Of the initial group in 2009, 145 patients had completed evaluations which included assessment of the Harris Hip Score and a radiographic assessment at a mean follow-up of 7.7 ± 2.2 years (range, 4 to 14 years). The rate of survival was calculated considering any reason for revision as failure. At follow-up, Harris hip score was 83.9 ± 16.1 (range, 21 to 100). Dislocation of the large articulation occurred in seven hips (4.8%), and only two recurred (1.4%) (one requiring additional revision). In addition, two intra-prosthetic dislocations of the small articulation (1.4%) were observed and needed revision surgery. The large number of earlier surgeries and non-union of the greater trochanter were related to recurrent instability. Two cups (1.4%) showed signs of definite loosening; six (4.1%) presented signs of possible loosening. Twenty-nine hips manifested femoral or acetabular osteolysis (20%), but only three were severe. Eight-year survival rate considering revision for any reason was 92.6% (95% CI, 85.5-96.4%). This series indicates that DM cups are a viable option to treat recurrent THA instability. Their design provides a low risk of recurrent instability without increasing mechanical complications.


Plasmon induced polymerization using a TERS approach: a platform for nanostructured 2D/1D material production.
Plasmon-induced chemical reactions have recently attracted great attention as a promising method for high efficiency light-energy conversion and proved to be useful in a wealth of different domains of chemistry and physics. One of the interesting and, so far, less explored avenues of such reactions is their potential for efficient, highly localized and controlled polymer production. Here, we present the first example of a localized, directed plasmon catalyzed polymerization process of a self-assembled monolayer on both silver and gold surfaces monitored by surface- and tip-enhanced Raman spectroscopy (SERS and TERS). As a proof-of-concept, a bi-functionalized dibenzo(1,2)dithiine-3,8-diamine (D3ATP) molecule that undergoes a well-known plasmon-induced coupling via the amino group into an azo group has been used. Initial dimerization is demonstrated using established marker bands associated with the formation of the azo group. A subsequent indicator for a polymerization reaction, the appearance of a new characteristic band, is monitored by time-dependent SERS and TERS experiments. We demonstrate that the dimerization reaction and hence, the subsequent polymerization, can be induced by a plasmonic feature, e.g. a TERS tip, at specific nanoscale locations and, at a much larger micron scale, by continuously scanning the plasmonic probe. The presented results provide the basis for designing further plasmonic catalysis experiments in general, and offer a new platform for producing ultra-thin polymer films with a defined structural dimension.


Overexpression of bone morphogenetic protein 10 in myocardium disrupts cardiac postnatal hypertrophic growth.
Postnatal cardiac hypertrophies have traditionally been classified into physiological or pathological hypertrophies. Both of them are induced by hemodynamic load. Cardiac postnatal hypertrophic growth is regarded as a part of the cardiac maturation process that is independent of the cardiac working load. However, the functional significance of this biological event has not been determined, mainly because of the difficulty in creating an experimental condition for testing the growth potential of functioning heart in the absence of hemodynamic load. Recently, we generated a novel transgenic mouse model (alphaMHC-BMP10) in which the cardiac-specific growth factor bone morphogenetic protein 10 (BMP10) is overexpressed in postnatal myocardium. These alphaMHC-BMP10 mice appear to have normal cardiogenesis throughout embryogenesis, but develop to smaller hearts within 6 weeks after birth. alphaMHC-BMP10 hearts are about half the normal size with 100% penetrance. Detailed morphometric analysis of cardiomyocytes clearly indicated that the compromised cardiac growth in alphaMHC-BMP10 mice was solely because of defect in cardiomyocyte postnatal hypertrophic growth. Physiological analysis further demonstrated that the responses of these hearts to both physiological (e.g. exercise-induced hypertrophy) and pathological hypertrophic stimuli remain normal. In addition, the alphaMHC-BMP10 mice develop subaortic narrowing and concentric myocardial thickening without obstruction by four weeks of age. Systematic analysis of potential intracellular pathways further suggested a novel genetic pathway regulating this previously undefined cardiac postnatal hypertrophic growth event. This is the first demonstration that cardiac postnatal hypertrophic growth can be specifically modified genetically and dissected out from physiological and pathological hypertrophies.


Comparison of combined antioxidants and thymoquinone in the prevention of testis ischemia - reperfusion injury.
We aimed to compare the preventive effects of combined antioxidants (CA1, 2) with a single antioxidant drug (thymoquinone; TQ) on experimental testis Ischemia/Reperfusion (I/R) injury. Thirty-five adult male Wistar rats were divided into five groups of seven rats each: control, testis I/R, testis I/R + CA1, testis I/R + CA2, and testis I/R + TQ. After 1 h of testicular ischemia, reperfusion was achieved by detorsion for 4 h. Antioxidants were intraperitoneally administered for 30 min prior to reperfusion. All rats were sacrificed 4 h after reperfusion to evaluate the tissue levels of malondialdehyde (MDA) and total antioxidant status (TAS) and the immunohistochemical evaluation of tissue inducible and endothelial nitric acid synthase (iNOS, eNOS) and apoptosis protease-activating factor 1 (APAF-1). MDA levels were lower and TAS values were higher in the I/R + antioxidant groups than in the I/R group (p < 0.05). iNOS and eNOS levels in the I/R + antioxidant groups were also lower than those in the I/R group (p < 0.05). There were no significant differences between the CA groups and the TQ group according to aforementioned parameters. In addition, tissue APAF-1 values were significantly higher in the I/R group than in the other groups. However, there was a significant difference between the TQ and CA groups in APAF-1 levels, which were highest in the TQ group (p < 0.05). Although TQ alone increased TAS values and reduced tissue iNOS and eNOS levels, combined antioxidant treatment may more effectively reduce apoptosis and increase preventive effects in testis I/R injury.