Can initial vaginal bleeding patterns in etonogestrel implant users predict subsequent bleeding in the first 2 years of use?
To evaluate if a simple method for characterizing vaginal bleeding patterns in etonogestrel contraceptive implant users can predict subsequent patterns and bleeding-related discontinuation over the first 2 years of use. We reanalyzed phase 3 study bleeding data for non-breastfeeding participants from the United States, Europe, Russia and Chile during the first 2 years of implant use to characterize and correlate bleeding patterns. We used 90-day reference periods with period 1.1 starting at Day 29 and ending at Day 118. We dichotomized bleeding patterns as "favorable" (amenorrhea, infrequent bleeding and normal frequency bleeding without prolonged bleeding) or "unfavorable' (prolonged and/or frequent bleeding) and tracked user groups based on these bleeding patterns in reference period 1.1 through Year 1 and from Year 1 through Year 2, respectively. We evaluated data from 537 and 428 women with up to 1 and 2 years use, respectively. Of the 325 (60.5%) women with favorable bleeding in reference period 1.1, 275 (84.6%) reported favorable bleeding also in reference period 2, 197 (60.6%) reported favorable bleeding throughout Year 1, and favorable bleeding in 75-85% of reference periods in Year 2. Among 212 (39.5%) women with unfavorable bleeding in reference period 1.1, 118 (55.7%) continued with unfavorable bleeding in reference period 2, while about 40%-50% reported favorable patterns in RP 2, 3 and/or 4. Initial favorable bleeding resulted in lower discontinuation rates than initial unfavorable bleeding in years 1 (3.7% vs 12.7%, p≪.0001) and 2 (2.5% vs 16.5%, p≪.0001). Implant users with favorable bleeding in the first reference period are likely to continue with favorable bleeding over the next 2 years. Initial bleeding patterns predict overall continuation rates in years 1 and 2. Implications Statement When evaluating vaginal bleeding in any 90-day reference period over 2 years of etonogestrel implant use, approximately 80% of women with favorable and 40% with unfavorable bleeding patterns will have favorable bleeding in the next reference periods. These findings can facilitate counseling regarding bleeding for women using the etonogestrel implant.


GRADE equity guidelines 2: considering health equity in GRADE guideline development: equity extension of the guideline development checklist.
To provide guidance for guideline developers on how to consider health equity at key stages of the guideline development process. Literature review followed by group discussions and consensus building. The key stages at which guideline developers could consider equity include setting priorities, guideline group membership, identifying the target audience(s), generating the guideline questions, considering the importance of outcomes and interventions, deciding what evidence to include and searching for evidence, summarizing the evidence and considering additional information, wording of recommendations, and evaluation and use. We provide examples of how guidelines have actually considered equity at each of these stages. Guideline projects should consider the aforementioned suggestions for recommendations that are equity sensitive.


Reversible calcitonin binding to solubilized sheep brain binding sites.
In this study we have solubilized and characterized binding sites for calcitonin (CT) from sheep brainstem. Autoradiography of 125I-labelled salmon CT (125I-sCT) binding to sheep diencephalon revealed a similar pattern of binding to that seen in other species, although the extent of distribution was greater in the sheep. CT binding activity could be extracted from membranes with either CHAPS or digitonin, but not with beta-octyl glucoside, 125I-sCT binding was saturable, with a dissociation constant for CHAPS-solubilized membranes of 2.8 +/- 0.5 nM and a maximum binding site concentration of 6.2 +/- 1.6 pmol/mg of protein. In competition binding studies, various CTs and their analogues demonstrated a similar rank order of potency to that seen in other CT receptor systems, Optimal binding occurred in the pH range 6.5-7.5, and was decreased in the presence of NaCl concentrations greater than 200 mM. In contrast with most other CT receptor binding systems, in which binding is poorly reversible, the binding of 125I-sCT to sheep brain binding sites underwent substantial dissociation upon addition of excess unlabelled sCT, with 40% and 46% dissociation after 2 h at 4 degree C in particulate and solubilized membranes respectively. Photoaffinity labelling of the binding site with the biologically active analogue 125I-[Arg11,18,4-azidobenzoyl-Lys14]sCT and analysis on SDS/PAGE under reducing conditions revealed a specific protein band of Mr approximately solubilized and particulate brain membranes. This is in accordance with the molecular size of CT receptors in other tissues where two species of receptor have been identified. one of Mr approximately 71,000 and another of Mr approximately 88,000. These results demonstrate the presence of high concentrations of CT binding sites in sheep brain which display different kinetic properties to those of CT receptors found in other tissues.


3D high throughput screening and profiling of embryoid bodies in thermoformed microwell plates.
3D organoids using stem cells to study development and disease are now widespread. These models are powerful to mimic in vivo situations but are currently associated with high variability and low throughput. For biomedical research, platforms are thus necessary to increase reproducibility and allow high-throughput screens (HTS). Here, we introduce a microwell platform, integrated in standard culture plates, for functional HTS. Using micro-thermoforming, we form round-bottom microwell arrays from optically clear cyclic olefin polymer films, and assemble them with bottom-less 96-well plates. We show that embryonic stem cells aggregate faster and more reproducibly (centricity, circularity) as compared to a state-of-the-art microwell array. We then run a screen of a chemical library to direct differentiation into primitive endoderm (PrE) and, using on-chip high content imaging (HCI), we identify molecules, including regulators of the cAMP pathway, regulating tissue size, morphology and PrE gene activity. We propose that this platform will benefit to the systematic study of organogenesis in vitro.


Probabilistic Machine Learning for Healthcare.
Machine learning can be used to make sense of healthcare data. Probabilistic machine learning models help provide a complete picture of observed data in healthcare. In this review, we examine how probabilistic machine learning can advance healthcare. We consider challenges in the predictive model building pipeline where probabilistic models can be beneficial, including calibration and missing data. Beyond predictive models, we also investigate the utility of probabilistic machine learning models in phenotyping, in generative models for clinical use cases, and in reinforcement learning.


Systemic isotretinoin treatment in a renal transplant patient developing sebaceous hyperplasia due to cyclosporine.
Sebaceous hyperplasia (SH) is a benign tumor with telangiectasia on it, yellowish or skin-colored, with papulosis. Besides genetic factors, aging, ultraviolet rays, sex hormones, calcineurin inhibitors, such as cyclosporin, tacrolimus and systemic steroids, play a role in the development of sebaceous hyperplasia. Cyclosporin is widely used in organ transplant patients. Acne, keratosis pilaris, sebaceous hyperplasia and epidermoid cysts, which are rare side effects, are frequently seen in renal transplant patients and it is suggested that the pilosebaceous unit develops as a result of occlusion with keratinous material. It is thought that cyclosporine causes these side effects by increasing the secretion of sebum and 5-alpha reductase enzyme activity. In this case study, wepresent here a 36-year-old female patient who had been on cyclosporine treatment for 25 years and had a large number of yellowish, umblike papules on her face for 20 years. She had been diagnosed with cyclosporin triggered by clinical and histopathological findings. The patient was started on 40 mg/day (0.6 mg/kg/day) systemic isotretinoin treatment, and after two months treatment, the patient had almost complete regression of the lesions. Systemic isotretinoin is effective and easy to treat treatment for patients with multiple lesions, especially when compared to other treatments. In addition, all of the cases reported in the literature are male, and this report presents the first female transplant patient with SH, which is induced by cyclosporine use.


Vitamin D status in a Brazilian cohort of adolescents and young adults with perinatally acquired human immunodeficiency virus infection.
The purpose was to determine the prevalence and related factors of vitamin D (VitD) insufficiency in adolescents and young adults with perinatally acquired human immunodeficiency virus. A cohort of 65 patients (17.6 ± 2 years) at the Federal University of Rio de Janeiro, Brazil, were examined for pubertal development, nutrition, serum parathormone and serum 25-hydroxyvitamin D [s25(OH)D]. s25(OH)D levels < 30 ng/mL (< 75 nmol/L) were defined as VitD insufficiency. CD4+ T-cell counts and viral load, history of worst clinical status, immunologic status as nadir, current immunologic status, and antiretroviral (ART) regimen were also evaluated as risk factors for VitD insufficiency. Mean s25(OH)D was 37.7 ± 13.9 ng/mL and 29.2% had VitD insufficiency. There was no difference between VitD status and gender, age, nutritional status, clinical and immunological classification, and type of ART. Only VitD consumption showed tendency of association with s25(OH)D (p = 0.064). Individuals analysed in summer/autumn season had a higher s25(OH)D compared to the ones analysed in winter/spring (42.6 ± 14.9 vs. 34.0 ± 11.9, p = 0.011). Although, the frequency of VitD insufficiency did not differ statistically between the groups (summer/autumn 17.9% vs. winter/spring 37.8%, p = 0.102), we suggest to monitor s25(OH)D in seropositive adolescents and young adults, especially during winter/spring months, even in sunny regions.


Coordination polymers built up from an s-tetrazine derived ligand and rare-earth ions: from a sequential dimensional expansion to organic-inorganic energy transfer.
Coordination polymers were prepared from a luminescent s-tetrazine ligand (OTz) and various rare-earth ions. Both the radii of the Ln ions and the synthetic conditions (especially pH) drive the formation of the product, and three crystalline phases were finally identified. Using a two-step process, it was possible to increase the dimensionality of one of these compounds from 1-D (MIL-165) to 2-D (MIL-166). The luminescence properties of these coordination polymers were studied. Whereas for Ln = La, Pr, Eu, Tb, the emission characteristic of the s-tetrazine core was observed, such emission was not detected for Ln = Nd, while the emission typical for the Nd ion was present, due to energy transfer from the tetrazine ligand to the Nd3+ ion.


Hyperpolarized magnetic resonance as a sensitive detector of metabolic function.
Hyperpolarized magnetic resonance allows for noninvasive measurements of biochemical reactions in vivo. Although this technique provides a unique tool for assaying enzymatic activities in intact organs, the scope of its application is still elusive for the wider scientific community. The purpose of this review is to provide key principles and parameters to guide the researcher interested in adopting this technology to address a biochemical, biomedical, or medical issue. It is presented in the form of a compendium containing the underlying essential physical concepts as well as suggestions to help assess the potential of the technique within the framework of specific research environments. Explicit examples are used to illustrate the power as well as the limitations of hyperpolarized magnetic resonance.


Stepwise engineering of a Pichia pastoris D-amino acid oxidase whole cell catalyst.
Trigonopsis variabilis D-amino acid oxidase (TvDAO) is a well characterized enzyme used for cephalosporin C conversion on industrial scale. However, the demands on the enzyme with respect to activity, operational stability and costs also vary with the field of application. Processes that use the soluble enzyme suffer from fast inactivation of TvDAO while immobilized oxidase preparations raise issues related to expensive carriers and catalyst efficiency. Therefore, oxidase preparations that are more robust and active than those currently available would enable a much broader range of economically viable applications of this enzyme in fine chemical syntheses. A multi-step engineering approach was chosen here to develop a robust and highly active Pichia pastoris TvDAO whole-cell biocatalyst. As compared to the native T. variabilis host, a more than seven-fold enhancement of the intracellular level of oxidase activity was achieved in P. pastoris through expression optimization by codon redesign as well as efficient subcellular targeting of the enzyme to peroxisomes. Multi copy integration further doubled expression and the specific activity of the whole cell catalyst. From a multicopy production strain, about 1.3 x 103 U/g wet cell weight (wcw) were derived by standard induction conditions feeding pure methanol. A fed-batch cultivation protocol using a mixture of methanol and glycerol in the induction phase attenuated the apparent toxicity of the recombinant oxidase to yield final biomass concentrations in the bioreactor of >or= 200 g/L compared to only 117 g/L using the standard methanol feed. Permeabilization of P. pastoris using 10% isopropanol yielded a whole-cell enzyme preparation that showed 49% of the total available intracellular oxidase activity and was notably stabilized (by three times compared to a widely used TvDAO expressing Escherichia coli strain) under conditions of D-methionine conversion using vigorous aeration. Stepwise optimization using a multi-level engineering approach has delivered a new P. pastoris whole cell TvDAO biocatalyst showing substantially enhanced specific activity and stability under operational conditions as compared to previously reported preparations of the enzyme. The production of the oxidase through fed-batch bioreactor culture and subsequent cell permeabilization is high-yielding and efficient. Therefore this P. pastoris catalyst has been evaluated for industrial purposes.


Improvement in Back and Leg Pain and Disability Following Adult Spinal Deformity Surgery: Study of 324 Patients With 2-year Follow-up and the Impact of Surgery on Patient-reported Outcomes.
A retrospective review of a prospective, multicenter adult spinal deformity (ASD) database. Our objective was to quantify the change in disability reported for patients with radiculopathy as compared with patients with back pain only following ASD realignment surgery. Studies utilizing patient-reported outcomes (PROs) have shown that ASD patients suffer from significant pain and disability. Although surgical correction has been effective at improving back and leg pain, no studies have investigated the impact of radiculopathy on pain and disability in ASD patients. Inclusion criteria were age ≥ 18 years and presence of spinal deformity as defined by coronal Cobb angle ≥ 20°, sagittal vertical axis (SVA) ≥ 5 cm, pelvic tilt (PT) angle ≥ 25°, or thoracic kyphosis (TK) angle ≥ 60°. Demographics and PRO were collected. Patients with radiculopathy were propensity matched with patients with back pain. Preoperative, postoperative, and 2-year follow-up radiographic parameters and PRO were analyzed. Three hundred twenty-four patients met inclusion criteria, and following propensity matching, 90 patients were placed into the radiculopathy and back pain groups. These groups showed no difference in demographic or radiographic parameters. The groups were similar in PRO, with a difference in leg pain as per design. At baseline, leg pain patients had higher disability [Oswestry Disability Index (ODI) and Scoliosis Research Society (SRS)]. Surgical strategies between the two groups showed no differences. Postoperative radiographic parameters showed no difference. Despite a significantly greater decrease in numerical rating scale (NRS) Leg for the leg pain group, postoperatively, these patients remained more disabled than the nonleg group in terms of NRS (back and leg), ODI, and most of the SRS domains. ASD patients with radiculopathy exhibit increased pain and disability when compared with patients without leg pain. This increased pain and disability persists after surgical correction in these patients. 3.


Dystrophin Analysis in Clinical Trials.
Duchenne muscular dystrophy (DMD) is a severe muscle-wasting disease with unmet medical need. The disease is caused by mutations that disrupt the open reading frame of the dystrophin protein that is required to maintain muscle fiber stability during contraction. Lacking dystrophin patients' muscle fibers are continuously damaged eventually leading to replacement of muscle tissue by fibrotic and adipose tissues and loss of muscle function. Many therapeutic approaches aiming at dystrophin restoration are in development, and some have been or are being tested in clinical trials. For these approaches, showing dystrophin restoration or increased dystrophin expression could serve as a pharmacodynamic biomarker to confirm mechanism of action. This review provides an overview of methods currently in use to assess dystrophin in clinical trial muscle biopsies and discusses challenges of dystrophin quantification and using dystrophin as a biomarker in clinical trials.


Tophaceous gout of the knee: revisiting MRI patterns in 30 patients.
To evaluate the locations and associated erosions of tophaceous gout in the knee and to reevaluate its characteristic magnetic resonance imaging features. We performed a retrospective review of the magnetic resonance (MR) images of the knees of 30 patients with histopathologically confirmed gouty tophi. Each tophaceous lesion was evaluated for location, morphology, signal intensity, and associated bone erosion or intraosseous involvement. The tophaceous masses were commonly located on the medial aspect of the infrapatellar fat pad and anterior joint recess (n = 28, 87%), the space adjacent to the lateral rim of the lateral femoral condyle (n = 25, 78%), and the intercondylar fossae (n = 22, 69%). All tophaceous lesions showed similar signal characteristics (low-intermediate signal intensities on T1-weighted images, with heterogeneous signal intensities on T2-weighted images), and 3 morphologic patterns were observed: amorphous masses (n = 27), linear crystalline-like deposits (n = 6), and cystic lesions of the bursae around the knee (n = 3). Associated bone erosions occurred in the lateral rim of the lateral femoral condyle (n = 8), the roof of the intercondylar notch (n = 7), the tibial eminence (n = 5), and the medial and lateral rim of the tibial plateau (n = 3). Intraosseous tophi were seen in the tibial plateau (n = 3) and patella (n = 1). Knowledge of the common locations and associated erosions of tophaceous gout and its characteristic MR appearance may facilitate its correct diagnosis with magnetic resonance imaging, particularly in patients with no clinical symptom or only an isolated lesion on MR images.


Ultrastructural characterisation of the proliferative (stem?) cells within the parenchyma of the normal "resting" breast.
In this study the proliferative (stem?) cells within the parenchyma of the normal "resting" breast were characterised by the ultrastructural examination of 60 mitotic cells. The parenchyma consists of epithelial and myoepithelial cells plus a few intraepithelial lymphocytes and macrophages. The majority of mitotic cells were randomly distributed throughout the lobules with a few present in ducts. In all cases the cells were identified as luminally positioned polarised epithelial cells. The proliferating cells had similar cytoplasmic features and were indistinguishable from adjacent interphase epithelial cells. No evidence was found for the division of subluminal epithelial or myoepithelial cells. These observations would be consistent with a single cell type giving rise to both epithelial and myoepithelial cells.


NAD(P)H, a directly operating antioxidant?
Endogenous oxygen- and nitrogen-centered free radicals are considered to play a decisive role in a variety of diseases such as neurodegenerative disorders, atherosclerosis, or cancer. Directly operating antioxidants limit the action of freely diffusing radicals by scavenging the attacking, oxidizing radical and re-reducing the oxidized biomolecule, i.e., the biomolecule-derived radical. From textbooks of biochemistry it is understood that NAD(P)H acts as a hydride (hydrogen anion) donor in a variety of enzymatic processes. One example is the re-reduction of GSSG to GSH, catalyzed by glutathione reductase. Because of this reaction, NADPH has been suggested to also act as an indirectly operating antioxidant, thus maintaining the antioxidative power of glutathione. To the best of our knowledge, however, neither NADPH nor NADH has been considered to be directly operating antioxidants. Based on recently published data, new experiments, and theoretical considerations, we propose that NAD(P)H represents a decisive, directly operating antioxidant that should be considered of major importance in the mitochondrial compartment. NAD(P)H fulfills this task both by scavenging toxic free radicals and repairing biomolecule-derived radicals.


Nonvolatile ternary resistive switching memory devices based on the polymer composites containing zinc oxide nanoparticles.
Nonvolatile ternary memory devices were fabricated from the composites polymer blends containing zinc oxide (ZnO) nanoparticles. When applying a negative bias on the top electrode, the fabricated devices with a simple sandwich structure of indium tin oxide (ITO)/composite polymer/aluminum (Al) exhibited three distinct resistance states, which could be labeled as "OFF", "ON1" and "ON2" for ternary data storage application. The ITO/polystyrene (PS) + ZnO/Al devices can endure 3 × 104 read-cycles and exhibit a retention time of 104 s. The resistance-temperature dependence at different resistance states was investigated to confirm the temperature-dependent properties. The resistance of the "OFF" and "ON1" state reveals negative temperature dependence, manifesting a typical semiconductor characteristic. The resistance of the "ON2" state exhibits positive temperature dependence, showing metallic properties.


Nonconscious emotional processing involves distinct neural pathways for pictures and videos.
Facial expressions are known to impact observers' behavior, even when they are not consciously identifiable. Relying on visual crowding, a perceptual phenomenon whereby peripheral faces become undiscriminable, we show that participants exposed to happy vs. neutral crowded faces rated the pleasantness of subsequent neutral targets accordingly to the facial expression's valence. Using functional magnetic resonance imaging (fMRI) along with psychophysiological interaction analysis, we investigated the neural determinants of this nonconscious preference bias, either induced by static (i.e., pictures) or dynamic (i.e., videos) facial expressions. We found that while static expressions activated primarily the ventral visual pathway (including task-related functional connectivity between the fusiform face area and the amygdala), dynamic expressions triggered the dorsal visual pathway (i.e., posterior partietal cortex) and the substantia innominata, a structure that is contiguous with the dorsal amygdala. As temporal cues are known to improve the processing of visible facial expressions, the absence of ventral activation we observed with crowded videos questions the capacity to integrate facial features and facial motions without awareness. Nevertheless, both static and dynamic facial expressions activated the hippocampus and the orbitofrontal cortex, suggesting that nonconscious preference judgments may arise from the evaluation of emotional context and the computation of aesthetic evaluation.


LASSO-Patternsearch algorithm with application to ophthalmology and genomic data.
The LASSO-Patternsearch algorithm is proposed to efficiently identify patterns of multiple dichotomous risk factors for outcomes of interest in demographic and genomic studies. The patterns considered are those that arise naturally from the log linear expansion of the multivariate Bernoulli density. The method is designed for the case where there is a possibly very large number of candidate patterns but it is believed that only a relatively small number are important. A LASSO is used to greatly reduce the number of candidate patterns, using a novel computational algorithm that can handle an extremely large number of unknowns simultaneously. The patterns surviving the LASSO are further pruned in the framework of (parametric) generalized linear models. A novel tuning procedure based on the GACV for Bernoulli outcomes, modified to act as a model selector, is used at both steps. We applied the method to myopia data from the population-based Beaver Dam Eye Study, exposing physiologically interesting interacting risk factors. We then applied the the method to data from a generative model of Rheumatoid Arthritis based on Problem 3 from the Genetic Analysis Workshop 15, successfully demonstrating its potential to efficiently recover higher order patterns from attribute vectors of length typical of genomic studies.


Global impact of tobacco control policies on smokeless tobacco use: a systematic review protocol.
Smokeless tobacco (ST) was consumed by 356 million people globally in 2017. Recent evidence shows that ST consumption is responsible for an estimated 652 494 all-cause deaths across the globe annually. The WHO Framework Convention on Tobacco Control (FCTC) was negotiated in 2003 and ratified in 2005 to implement effective tobacco control measures. While the policy measures enacted through various tobacco control laws have been effective in reducing the incidence and prevalence of smoking, the impact of ST-related policies (within WHO FCTC and beyond) on ST use is under-researched and not collated. A systematic review will be conducted to collate all available ST-related policies implemented across various countries and assess their impact on ST use. The following databases will be searched: Medline, EMBASE, PsycINFO, Cumulative Index to Nursing and Allied Health Literature, Scopus, EconLit, ISI Web of Science, Cochrane Library (CENTRAL), African Index Medicus, LILACS, Scientific Electronic Library Online, Index Medicus for the Eastern Mediterranean Region, Index Medicus for South-East Asia Region, Western Pacific Region Index Medicus and WHO Library Database, as well as Google search engine and country-specific government websites. All ST-related policy documents (FCTC and non-FCTC) will be included. Results will be limited to literature published since 2005 in English and regional languages (Bengali, Hindi and Urdu). Two reviewers will independently employ two-stage screening to determine inclusion. The Effective Public Health Practice Project's 'Quality Assessment Tool for Quantitative Studies' will be used to record ratings of quality and risk of bias among studies selected for inclusion. Data will be extracted using a standardised form. Meta-analysis and narrative synthesis will be used. Permission for ethics exemption of the review was obtained from the Centre for Chronic Disease Control's Institutional Ethics Committee, India (CCDC_IEC_06_2020; dated 16 April 2020). The results will be disseminated through publications in a peer-reviewed journal and will be presented in national and international conferences. CRD42020191946.


Membrane stiffness is modified by integral membrane proteins.
The ease with which a cell membrane can bend and deform is important for a wide range of biological functions. Peripheral proteins that induce curvature in membranes (e.g. BAR domains) have been studied for a number of years. Little is known, however, about the effect of integral membrane proteins on the stiffness of a membrane (characterised by the bending rigidity, Kc). We demonstrate by computer simulation that adding integral membrane proteins at physiological densities alters the stiffness of the membrane. First we establish that the coarse-grained MARTINI forcefield is able to accurately reproduce the bending rigidity of a small patch of 1500 phosphatidyl choline lipids by comparing the calculated value to both experiment and an atomistic simulation of the same system. This enables us to simulate the dynamics of large (ca. 50 000 lipids) patches of membrane using the MARTINI coarse-grained description. We find that altering the lipid composition changes the bending rigidity. Adding integral membrane proteins to lipid bilayers also changes the bending rigidity, whilst adding a simple peripheral membrane protein has no effect. Our results suggest that integral membrane proteins can have different effects, and in the case of the bacterial outer membrane protein, BtuB, the greater the density of protein, the larger the reduction in stiffness.


Laser-induced ultrasonics for detection of low-amplitude grating through metal layers with finite roughness.
We report on the use of laser-induced ultrasonics for the detection of gratings with amplitudes as small as 0.5 nm, buried underneath an optically opaque nickel layer. In our experiments, we use gratings fabricated on top of a nickel layer on glass, and we optically pump and probe the sample from the glass side. The diffraction of the probe pulse from the acoustic echo from the buried grating is measured as a function of time. We use a numerical model to show how the various physical phenomena such as interface displacement, strain-optic effects, thermo-optic effects, and surface roughness influence the shape and strength of the time-dependent diffraction signal. More importantly, we use a Rayleigh-Rice scattering theory to quantify the amount of light scattering, which is then used as in input parameter in our numerical model to predict the time-dependent diffracted signal.


Four lessons in adaptive leadership.
The armed services have been in the business of leadership development much longer than the corporate world has. Today's military leaders need tools and techniques to face a fast-changing and unpredictable type of enemy--so the armed services train their officers in ways that build a culture of readiness and commitment. Business leaders need to foster an adaptive culture to survive and succeed, given that they, too, face unprecedented uncertainty--and new types of competitors. Michael Useem and his colleagues at the Wharton School incorporate exposure to military leadership into MBA and executive MBA programs. Highlights include direct contact in the classroom with leaders in the U.S. Army, the U.S. Marine Corps, and the Department of Defense, along with field-training exercises and battlefield visits. The programs are designed to help students connect viscerally to essential leadership lessons. Four are featured in the article: Meet the troops. Creating a personal link is crucial to leading people in challenging times. Make decisions. Making good and timely calls is the crux of leadership. Mission first. Focus on common purpose and eschew personal gain. Convey strategic intent. Make the objectives clear, but give people the freedom to execute on them in their own way.


The glucagon-producing alpha cell: an electrophysiologically exceptional cell.
Activation of potassium channels normally serves to reduce cellular activity but recent data indicate that the glucagon-secreting alpha cells are different in this respect and that inhibition of voltage-gated potassium channels results in a paradoxical inhibition of glucagon secretion. Here we discuss these findings and attempt to provide a model for the regulation of glucagon secretion that incorporates these observations.


Functional diversification of the chemical landscapes of yeast Sec14-like phosphatidylinositol transfer protein lipid-binding cavities.
Phosphatidylinositol-transfer proteins (PITPs) are key regulators of lipid signaling in eukaryotic cells. These proteins both potentiate the activities of phosphatidylinositol (PtdIns) 4-OH kinases and help channel production of specific pools of phosphatidylinositol 4-phosphate (PtdIns(4)P) dedicated to specific biological outcomes. In this manner, PITPs represent a major contributor to the mechanisms by which the biological outcomes of phosphoinositide are diversified. The two-ligand priming model proposes that the engine by which Sec14-like PITPs potentiate PtdIns kinase activities is a heterotypic lipid-exchange cycle where PtdIns is a common exchange substrate among the Sec14-like PITP family, but the second exchange ligand varies with the PITP. A major prediction of this model is that second-exchangeable ligand identity will vary from PITP to PITP. To address the heterogeneity in the second exchange ligand for Sec14-like PITPs, we used structural, computational, and biochemical approaches to probe the diversities of the lipid-binding cavity microenvironments of the yeast Sec14-like PITPs. The collective data report that yeast Sec14-like PITP lipid-binding pockets indeed define diverse chemical microenvironments that translate into differential ligand-binding specificities across this protein family.


Plasma pyridoxal-5-phosphate is inversely associated with systemic markers of inflammation in a population of U.S. adults.
Low vitamin B-6 status, based on plasma concentrations of pyridoxal-5-phosphate (PLP), has been identified in inflammatory diseases, including cardiovascular disease, rheumatoid arthritis, inflammatory bowel disease, and diabetes. Our objective was to examine the association between plasma PLP and multiple markers of inflammation in a community-based cohort [n = 2229 participants (55% women, mean age 61 ± 9 y)]. We created an overall inflammation score (IS) as the sum of standardized values of 13 individual inflammatory markers. Multivariable-adjusted regression analysis was used to assess the associations between the IS and plasma PLP. Geometric mean plasma PLP concentrations were lower in the highest tertile category of IS relative to the lowest (61 vs. 80 nmol/L; P-trend < 0.0001). Similarly, the prevalence of PLP insufficiency was significantly higher for participants in the highest compared with the lowest tertiles for IS categories. These relationships persisted after accounting for vitamin B-6 intake. Also, there were significant inverse relationships between plasma PLP and 4 IS based on functionally related markers, including acute phase reactants, cytokines, adhesion molecules, and oxidative stress. In addition, secondary analyses revealed that many of the individual inflammatory markers were inversely associated with plasma PLP after adjusting for plasma C-reactive protein concentration. This study, in combination with past findings, further supports our hypothesis that inflammation is associated with a functional deficiency of vitamin B-6. We discuss 2 possible roles for PLP in the inflammatory process, including tryptophan metabolism and serine hydroxymethyltransferase activity.


Nociceptive afferent vagal input is enhanced after transection of the aortic depressor nerve.
To test the hypothesis that baroreceptor reflexes are involved in the reduced nociceptive responses associated with hypertension, the effects of acute intravenous doses of serotonin (0.75-144 micrograms/kg) on inhibition of the tail-flick reflex, blood pressure, and heart rate were examined in lightly pentobarbital-anesthetized Sprague-Dawley, Wistar-Kyoto, and spontaneously hypertensive rats before and after bilateral transection of the aortic depressor nerve. Before transection, the ED50s for inhibition of the tail-flick reflex in the three strains of rats were 14, 13, and 44 micrograms/kg, respectively. After bilateral transection of the aortic depressor nerve, all three strains displayed a significant increase in sensitivity toward the serotonin-induced inhibition of the tail-flick reflex (ED50s of 7, 7, and 6 micrograms/kg, respectively). There were no changes in the cardiovascular responses to serotonin after transection of the aortic depressor nerve in Wistar-Kyoto or spontaneously hypertensive rats. In Sprague-Dawley rats, however, the cardio-pulmonary cardiovascular afferent-mediated responses were enhanced. In Sprague-Dawley rats, the nociceptive sensitivity to serotonin was unaltered when administered during either the peak increase or decrease in pressure produced by intravenous phenylephrine or nitroprusside, respectively. These results suggest that 1) afferent fibers within the aortic depressor nerve provide strong tonic inhibitory influences on the noxious information conveyed by the vagus in response to intravenous serotonin and 2) these fibers appear to produce their effects by mechanisms that are unrelated to baroreceptor function.


Cigarette smoking, TP53 Arg72Pro, TP53BP1 Asp353Glu and the risk of lung cancer in a Japanese population.
The tumor suppressor protein p53 (TP53) plays a central role in directing cellular responses to DNA damage. Tumor protein 53-binding protein 1 (TP53BP1) binds to TP53 and has a potential role in DNA damage responses. DNA damage-dependent interaction between TP53 and TP53BP1 may contribute to lung cancer risk. We aimed to assess whether or not TP53 and TP53BP1 genetic polymorphisms modulate lung cancer susceptibility in a Japanese population. We investigated the relationship of the TP53 Arg72Pro and TP53BP1 Asp353Glu polymorphisms to lung cancer risk with special reference to polymorphism-polymorphism and polymorphism-smoking interactions among 462 lung cancer cases and 379 controls. The Glu/Glu genotype of TP53BP1 Asp353Glu polymorphism was associated with a decreased risk of lung cancer [odds ratio (OR) = 0.46, 95% confidence interval (CI) = 0.29-0.74]. There was no polymorphism-smoking interaction. A combination of the Pro allele carriage of the TP53 Arg72Pro polymorphism and the Glu/Glu genotype of the TP53BP1 Asp353Glu polymorphism was associated with a decreased risk of lung cancer (OR=0.38, 95% CI=0.17-0.83). The multiplicative interaction measure was statistically significant (OR for interaction = 2.93, 95% CI=1.24-6.93). The relative excess risk due to interaction and attributable proportion due to interaction were 0.74 (95% CI=0.38-1.20) and 0.63 (95% CI=0.05-1.21), respectively. Both the additive interaction measures were not equal to zero, suggesting that the existence of a biological interaction. Our findings indicate the possible association of the Glu allele of the TP53BP1 Asp353Glu polymorphism with lower risk of lung cancer especially among the Pro allele carriers of the TP53 Arg72Pro polymorphism.


Eveningness is associated with poor sleep quality and negative affect in obsessive-compulsive disorder.
Background Obsessive-compulsive disorder (OCD) is characterized by intrusive thoughts and repetitive behaviors that severely encumber daily functioning. OCD patients seem to exhibit sleep disturbances, especially delayed bedtimes that reflect disrupted circadian rhythmicity. Morningness-eveningness is a fundamental factor reflecting individual variations in diurnal preferences related to sleep and waking activities. Eveningness reflecting a delayed sleep-wake timing has repeatedly been associated with sleep problems and negative affect (NA). Therefore, the aim of this study was to examine the associations between morningness-eveningness, sleep complaints, and symptom severity in OCD patients and compared with a mixed psychiatric control group. Materials and methods The data of 49 OCD and 49 mixed psychiatric inpatients (with unipolar depression and anxiety disorders) were analyzed. Patients completed questionnaires regarding morningness-eveningness, sleep quality, nightmare frequency, depression, anxiety, and affective states. Obsessive and compulsive symptom severity was also assessed within the OCD group by clinician-rated scales. Results Eveningness preference was associated with impaired sleep quality and higher NA in OCD patients. In addition, impaired sleep quality showed a moderate correlation with anxiety and strong correlations with depressive symptoms and NA. Interestingly, in the mixed psychiatric group, eveningness was not linked to NA, and sleep quality also showed weaker associations with depressive symptoms and NA. Within the OCD group, eveningness preference was predictive of poorer sleep quality regardless the influence of depressive symptoms. Conclusion Our findings suggest that eveningness and sleep complaints are predictive of affective dysfunctions, and should be carefully considered in the evaluation and treatment of OCD patients.


Inhibition of antigen-specific T lymphocyte activation by structurally related Ir gene-controlled polymers. II. Competitive inhibition of I-E-restricted, antigen-specific T cell responses.
Our previous studies have defined a highly specific competitive inhibition between a pair of structurally related antigens (GT and GAT) for antigen presentation by accessory cells. The present report investigates this phenomenon in a second antigenic system, which is controlled by a distinct Ir gene product. Two GL phi-specific, I-Ed-restricted, interleukin 2-producing T cell hybridomas were constructed. The antigenic fine specificity of these two hybrid clones was distinct. One hybrid reacted solely with GL phi while the second cross-reacted with GLleu and GLT. These latter two copolymers, as well as the antigen GL, were found to inhibit the GL phi response of the non-cross-reactive hybrid. The structurally related antigen G phi was not inhibitory for this clone's response. The cross-reactive GL phi hybrid could also be inhibited, but, in this case, G phi and not GL caused the inhibition. Reciprocal inhibitions could be demonstrated between these and other hybrids (e.g., GAT responsive), indicating a very high degree of specificity to the inhibition. The inhibition caused by the various copolymers was reversible by increasing the concentration of GL phi, This effect was localized to the antigen-presenting cell and not the T cell hybridoma. Functionally, this competition did not appear to be for antigen uptake or general antigen processing. These findings generalize the phenomenon of antigen competition to a second antigen system in the context of a second Ia molecule. The possible mechanisms accounting for the complex pattern of specificities in this system are discussed.


Operational modes of a ferroelectric LCoS modulator for displaying binary polarization, amplitude, and phase diffraction gratings.
We analyze the performance of a ferroelectric liquid crystal on silicon display (FLCoS) as a binary polarization diffraction grating. We analyze the correspondence between the two polarization states emerging from the displayed grating and the polarization and intensity of the diffracted orders generated at the Fourier diffraction plane. This polarization-diffraction analysis leads, in a simple manner, to configurations yielding binary amplitude or binary phase modulation by incorporating an analyzer on the reflected beam. Based on this analysis, we present two useful variations of the polarization configuration. The first is a simplification using a single polarizer, which provides equivalent results for amplitude or phase modulation as the more general operational mode involving two polarizers. The second variation is proposed to compensate the reduction of the diffraction efficiency when the operating wavelength differs from the design one (for which the FLCoS liquid-crystal layer acts as a half-wave plate). In this situation we show how the ideal grating performance can be recovered in spite of the phase-shift mismatch originated by chromatic dispersion. In all cases, we provide experimental results that verify the theoretical analyses.


A composite MyD88/CD40 switch synergistically activates mouse and human dendritic cells for enhanced antitumor efficacy.
The in vivo therapeutic efficacy of DC-based cancer vaccines is limited by suboptimal DC maturation protocols. Although delivery of TLR adjuvants systemically boosts DC-based cancer vaccine efficacy, it could also increase toxicity. Here, we have engineered a drug-inducible, composite activation receptor for DCs (referred to herein as DC-CAR) comprising the TLR adaptor MyD88, the CD40 cytoplasmic region, and 2 ligand-binding FKBP12 domains. Administration of a lipid-permeant dimerizing ligand (AP1903) induced oligomerization and activation of this fusion protein, which we termed iMyD88/CD40. AP1903 administration to vaccinated mice enabled prolonged and targeted activation of iMyD88/CD40-modified DCs. Compared with conventionally matured DCs, AP1903-activated iMyD88/CD40-DCs had increased activation of proinflammatory MAPKs. AP1903-activated iMyD88/CD40-transduced human or mouse DCs also produced higher levels of Th1 cytokines, showed improved migration in vivo, and enhanced both antigen-specific CD8+ T cell responses and innate NK cell responses. Furthermore, treatment with AP1903 in vaccinated mice led to robust antitumor immunity against preestablished E.G7-OVA lymphomas and aggressive B16.F10 tumors. Thus, the iMyD88/CD40 unified "switch" effectively and safely replaced exogenous adjuvant cocktails, allowing remote and sustained DC activation in vivo. DC "licensing" through iMyD88/CD40 may represent a mechanism by which to exploit the natural synergy between the TLR and CD40 signaling pathways in DCs using a single small molecule drug and could augment the efficacy of antitumor DC-based vaccines.


Great flights by great snipes: long and fast non-stop migration over benign habitats.
Migratory land birds perform extreme endurance flights when crossing ecological barriers, such as deserts, oceans and ice-caps. When travelling over benign areas, birds are expected to migrate by shorter flight steps, since carrying the heavy fuel loads needed for long non-stop flights comes at considerable cost. Here, we show that great snipes Gallinago media made long and fast non-stop flights (4300-6800 km in 48-96 h), not only over deserts and seas but also over wide areas of suitable habitats, which represents a previously unknown migration strategy among land birds. Furthermore, the great snipes achieved very high ground speeds (15-27 m s(-1)), which was not an effect of strong tailwind support, and we know of no other animal that travels this rapidly over such a long distance. Our results demonstrate that some migratory birds are prepared to accept extreme costs of strenuous exercise and large fuel loads, even when stopover sites are available along the route and there is little tailwind assistance. A strategy of storing a lot of energy before departure, even if migration is over benign habitats, may be advantageous owing to differential conditions of fuel deposition, predation or infection risk along the migration route.


Red blood cell homeostasis in children and adults with and without asymptomatic malaria infection in Burkina Faso.
Asymptomatic malaria infections may affect red blood cell (RBC) homeostasis. Reports indicate a role for chronic hemolysis and splenomegaly, however, the underlying processes are incompletely understood. New hematology analysers provide parameters for a more comprehensive analysis of RBC hemostasis. Complete blood counts were analysed in subjects from all age groups (n = 1118) living in a malaria hyperendemic area and cytokines and iron biomarkers were also measured. Subjects were divided into age groups (<2 years, 2-4, 5-14 and ≥15 years old) and clinical categories (smear-negative healthy subjects, asymptomatic malaria and clinical malaria). We found that hemoglobin levels were similar in smear-negative healthy children and asymptomatic malaria children but significantly lower in clinical malaria with a maximum difference of 2.2 g/dl in children <2 years decreasing to 0.1 g/dl in those aged ≥15 years. Delta-He, presenting different hemoglobinization of reticulocytes and RBC, levels were lower in asymptomatic and clinial malaria, indicating a recent effect of malaria on erythropoiesis. Reticulocyte counts and reticulocyte production index (RPI), indicating the erythropoietic capacity of the bone marrow, were higher in young children with malaria compared to smear-negative subjects. A negative correlation between reticulocyte counts and Hb levels was found in asymptomatic malaria (ρ = -0.32, p<0.001) unlike in clinical malaria (ρ = -0.008, p = 0.92). Free-Hb levels, indicating hemolysis, were only higher in clinical malaria. Phagocytozing monocytes, indicating erythophagocytosis, were highest in clinical malaria, followed by asymptomatic malaria and smear-negative subjects. Circulating cytokines and iron biomarkers (hepcidin, ferritin) showed similar patterns. Pro/anti-inflammatory (IL-6/IL-10) ratio was higher in clinical than asymptomatic malaria. Cytokine production capacity of ex-vivo whole blood stimulation with LPS was lower in children with asymptomatic malaria compared to smear-negative healthy children. Bone marrow response can compensate the increased red blood cell loss in asymptomatic malaria, unlike in clinical malaria, possibly because of limited level and length of inflammation. Trial registration: Prospective diagnostic study: ClinicalTrials.gov identifier: NCT02669823. Explorative cross-sectional field study: ClinicalTrials.gov identifier: NCT03176719.


Localized soft elasticity in liquid crystal elastomers.
Synthetic approaches to prepare designer materials that localize deformation, by combining rigidity and compliance in a single material, have been widely sought. Bottom-up approaches, such as the self-organization of liquid crystals, offer potential advantages over top-down patterning methods such as photolithographic control of crosslink density, relating to the ease of preparation and fidelity of resolution. Here, we report on the directed self-assembly of materials with spatial and hierarchical variation in mechanical anisotropy. The highly nonlinear mechanical properties of the liquid crystalline elastomers examined here enables strain to be locally reduced >15-fold without introducing compositional variation or other heterogeneities. Each domain (⩾0.01 mm(2)) exhibits anisotropic nonlinear response to load based on the alignment of the molecular orientation with the loading axis. Accordingly, we design monoliths that localize deformation in uniaxial and biaxial tension, shear, bending and crack propagation, and subsequently demonstrate substrates for globally deformable yet locally stiff electronics.


Educational attainment and work disability in cancer survivors: Do diagnosis and comorbidity affect this association?
To study whether educational attainment had less impact on work disability in cancer survivors than in individuals without cancer. To study whether comorbidity had a higher impact on work disability in low-educated cancer survivors than in high-educated and whether this impact differed when compared with individuals without cancer. Linkage of population-based public health survey data and the Danish Cancer Registry formed two groups: cancer survivors (n = 3,514) and cancer-free individuals (n = 171,262). In logistic regression models, the risk of experiencing an 8-week sick leave spell and the granting of disability pension within a 3-year follow-up period was studied in three educational levels and whether these associations were modified by history of cancer and comorbidity. Odds ratios (OR) with 95% confidence intervals (CI) are reported. Non-stratified adjusted risk of experiencing an 8-week sick leave spell (OR: 1.41, 95% CI (1.33-1.49)) or being granted a disability pension (OR: 1.61, 95% CI (1.31-1.97)) was significantly higher in low-educated than in high-educated respondents. Cancer or comorbidity did not significantly interact with education on the risk of work disability. A moderate impact of low education on future work disability was found for all respondents, neither history of cancer nor comorbidity modified this association.


Transcatheter embolization in pediatric blunt renal trauma: Case report and review of the literature.
This study aims to evaluate the clinical use and outcomes of transcatheter embolization in the setting of pediatric blunt renal trauma cases using an index case and review of the literature. Although the treatment method selection depends on the grade of the trauma and the hemodynamıc status of the patient, conservative methods are considered initially in the setting of pediatric blunt renal trauma. Transcatheter embolization, which is a relatively conservative treatment option, is commonly utilized in adult blunt renal trauma patient population; however, experience in the pediatric population is scarce. A seven-year-old male patient was admitted due to grade IV renal injury secondary to blunt abdominal trauma. He was conservatively followed until -on the post-trauma day 15- he developed gross hematuria and an unstable hemodynamic status. A renal angiogram was performed, which revealed right-sided renal inferior segmental artery pseudoaneurysm and arterio-venous fistula. Subsequently, a transcatheter embolization was performed. There were no complications and no permanent renal injury. Review of the literature revealed that the success rate of transcatheter embolization is 89.47% in the setting of pediatric blunt abdominal trauma-related renal injury regardless of the renal trauma grade. Four patients underwent nephrectomy due to a failed transcatheter embolization procedure. There are not any patients who suffered from embolization-related complications. These results suggest that transcatheter embolization can be safely performed in children with renal injuries due to blunt abdominal trauma with high success and low complication rates and preservation of renal function.


TLR13 recognizes bacterial 23S rRNA devoid of erythromycin resistance-forming modification.
Host protection from infection relies on the recognition of pathogens by innate pattern-recognition receptors such as Toll-like receptors (TLRs). Here, we show that the orphan receptor TLR13 in mice recognizes a conserved 23S ribosomal RNA (rRNA) sequence that is the binding site of macrolide, lincosamide, and streptogramin group (MLS) antibiotics (including erythromycin) in bacteria. Notably, 23S rRNA from clinical isolates of erythromycin-resistant Staphylococcus aureus and synthetic oligoribonucleotides carrying methylated adenosine or a guanosine mimicking a MLS resistance-causing modification failed to stimulate TLR13. Thus, our results reveal both a natural TLR13 ligand and specific mechanisms of antibiotic resistance as potent bacterial immune evasion strategy, avoiding recognition via TLR13.


Peripheral inhibition of glutaminase reduces carrageenan-induced Fos expression in the superficial dorsal horn of the rat.
In inflamed tissue, the levels of the excitatory amino acid glutamate are increased. Glutamate sensitizes peripheral axons of primary afferent neurons during inflammation leading to decreased firing threshold and hyperexcitability. One proposed source of glutamate is the primary afferent. Antagonizing glutamate receptors on peripheral axons of primary afferents during inflammation provides analgesia in animals and humans. The enzyme glutaminase is used by primary sensory neurons to convert glutamine to glutamate, and peripheral inhibition of glutaminase with 6-diazo-5-oxo-l-norleucine (DON) provides long-lasting analgesia during inflammation. In this study, we measured the effects of glutaminase inhibition on carrageenan-induced spinal Fos expression. Rats were given intraplantar injections of carrageenan and treated locally with either vehicle or DON. After 3h of inflammation, hind paw swelling and spinal expression of Fos were examined. CellProfiler was used to automate Fos nuclei counting in five laminar groupings in the spinal cord (I-II, III-IV, V-VI, VII-IX, X). Carrageenan increased hind paw thickness by approximately 70% and spinal Fos expression in superficial (I-II) and deep (V-VI) laminae by 10-fold and 5-fold, respectively. Treatment with DON reduced hind paw swelling by approximately 13% and suppressed Fos expression in the laminae I-II by approximately 54%, but not the deep laminae. Our results further support the notion of glutamate as a peripheral inflammatory mediator and indicate that glutaminase should be considered as a novel therapeutic target for treatment of inflammatory pain.


Morphological alterations within the peripheral fixation of the iris dilator muscle in eyes with pigmentary glaucoma.
To analyze the peripheral fixation of the iris dilator muscle in normal eyes and in eyes with pigmentary glaucoma (PG). Using 63 control eyes (age 18 months-99 years), the peripheral iris dilator was investigated by light microscopy, immunohistochemistry, and electron microscopy. Development was studied using 18 differently aged fetal eyes stained immunohistochemically against α-smooth muscle (SM) actin. The peripheral iris dilator muscle in PG was analyzed using semithin and ultrathin sections of six glutaraldehyde-fixed eyes from three donors aged 38, 62, and 74 years. In normal eyes, the peripheral end of the iris dilator muscle is arranged in a sphincter-like manner. Arcade-shaped tendinous connections associated with myofibroblasts (iridial strands) anchor the iris dilator within the elastic-fibromuscular ciliary meshwork that also serves as fixation area for the elastic tendons of the inner ciliary muscle portions. The iridial strands are innervated and can adapt their length during accommodation. The PG eyes show incomplete circular bundles and iridial strands that are mainly anchored to the iris stroma and the flexible uveal parts of the trabecular meshwork. The normal anchorage of the peripheral iris dilator and its presumably neuronally regulated length adaptation stabilize the peripheral iris during accommodation. Insufficient fixation in PG could promote posterior bowing of the iris with rubbing against the zonular fibers and pigment liberation from the iris pigmented epithelium.


Novel pleiotropic risk loci for melanoma and nevus density implicate multiple biological pathways.
The total number of acquired melanocytic nevi on the skin is strongly correlated with melanoma risk. Here we report a meta-analysis of 11 nevus GWAS from Australia, Netherlands, UK, and USA comprising 52,506 individuals. We confirm known loci including MTAP, PLA2G6, and IRF4, and detect novel SNPs in KITLG and a region of 9q32. In a bivariate analysis combining the nevus results with a recent melanoma GWAS meta-analysis (12,874 cases, 23,203 controls), SNPs near GPRC5A, CYP1B1, PPARGC1B, HDAC4, FAM208B, DOCK8, and SYNE2 reached global significance, and other loci, including MIR146A and OBFC1, reached a suggestive level. Overall, we conclude that most nevus genes affect melanoma risk (KITLG an exception), while many melanoma risk loci do not alter nevus count. For example, variants in TERC and OBFC1 affect both traits, but other telomere length maintenance genes seem to affect melanoma risk only. Our findings implicate multiple pathways in nevogenesis.


Pain in aquatic animals.
Recent developments in the study of pain in animals have demonstrated the potential for pain perception in a variety of wholly aquatic species such as molluscs, crustaceans and fish. This allows us to gain insight into how the ecological pressures and differential life history of living in a watery medium can yield novel data that inform the comparative physiology and evolution of pain. Nociception is the simple detection of potentially painful stimuli usually accompanied by a reflex withdrawal response, and nociceptors have been found in aquatic invertebrates such as the sea slug Aplysia. It would seem adaptive to have a warning system that allows animals to avoid life-threatening injury, yet debate does still continue over the capacity for non-mammalian species to experience the discomfort or suffering that is a key component of pain rather than a nociceptive reflex. Contemporary studies over the last 10 years have demonstrated that bony fish possess nociceptors that are similar to those in mammals; that they demonstrate pain-related changes in physiology and behaviour that are reduced by painkillers; that they exhibit higher brain activity when painfully stimulated; and that pain is more important than showing fear or anti-predator behaviour in bony fish. The neurophysiological basis of nociception or pain in fish is demonstrably similar to that in mammals. Pain perception in invertebrates is more controversial as they lack the vertebrate brain, yet recent research evidence confirms that there are behavioural changes in response to potentially painful events. This review will assess the field of pain perception in aquatic species, focusing on fish and selected invertebrate groups to interpret how research findings can inform our understanding of the physiology and evolution of pain. Further, if we accept these animals may be capable of experiencing the negative experience of pain, then the wider implications of human use of these animals should be considered.


Role of vasopressin in response to intrarenal infusions of alpha-2 adrenoceptor agonists.
Previous studies have indicated that the effects of renal alpha-2 adrenoceptor stimulation are mediated through the blockade of the renal effects of vasopressin. If this premise is correct then 1) specific antagonists of the antidiuretic effect of vasopressin (V2 antagonists) should mimic alpha-2 adrenoceptor stimulation and 2) in the presence of V2 antagonists, the diuretic and natriuretic effect of clonidine should be attenuated. The renal effects of [d(CH2)5,D-Ile2,Ile4]AVP, a specific V2 antagonist, were studied. On the day of the experiment, uninephrectomized rats were anesthetized, and the carotid artery and jugular vein were cannulated for recording blood pressure and saline infusion, respectively. The left kidney was exposed and the ureter cannulated. A 31-gauge needle was advanced into the renal artery to permit direct i.r. infusion of study drugs. Bolus doses of the V2 antagonist (0, 1, 3, 10, or 30 nmol/kg i.v.) produced a dose-related increase in urine volume and free water clearance at all doses tested. Sodium excretion increased only at the higher doses (10 and 30 nmol/kg). This dose-related dissociation in water and then sodium excretion is similar to that observed after i.r. clonidine infusions. In the presence of the V2 antagonist, clonidine (3 micrograms/kg/min) had no effect on urine volume or free water clearance but significantly decreased the excretion of sodium from control. These results demonstrate that V2 antagonists mimic the effects of i.r. clonidine. As well, in the absence of vasopressin (V2 antagonism), the effects of clonidine are attenuated. Moreover, they are also consistent with not only an antidiuretic role for endogenous vasopressin but also an antinatriuretic one.


Spontaneous hemoperitoneum in cats: 65 cases (1994-2006).
To describe the clinical signs, physical examination findings, clinical laboratory abnormalities, etiology, and outcome in cats with spontaneous hemoperitoneum. Retrospective case series. 65 client-owned cats with spontaneous hemoperitoneum. Medical records of cats with spontaneous hemoperitoneum at 7 large referral clinics were reviewed. Cats were included if a definitive diagnosis of spontaneous hemoperitoneum could be obtained from review of the medical records. 65 cats met inclusion criteria. The most common historical findings were lethargy, anorexia, and vomiting. Common findings on physical examination included inadequate hydration status and hypothermia. The most common clinicopathologic abnormalities were high serum AST activity, anemia, prolonged prothrombin time, and prolonged partial thromboplastin time. Forty-six percent (30/65) of cats had abdominal neoplasia, and 54% (35/65) had nonneoplastic conditions. Hemangiosarcoma was the most often diagnosed neoplasm (18/30; 60%), and the spleen was the most common location for neoplasia (11/30; 37%). Eight cats survived to be discharged from the hospital. Cats with neoplasia were significantly older and had significantly lower PCVs than cats with non-neoplastic disease. Spontaneous hemoperitoneum in cats often results in debilitating clinical consequences. In contrast to dogs with hemoperitoneum, the cause of hemoperitoneum in cats is approximately evenly distributed between neoplastic and nonneoplastic diseases. Although only a few cats were treated in this study, the prognosis appears poor.


Gamma-aminobutyric acid (GABA) is an autocrine excitatory transmitter in human pancreatic beta-cells.
Paracrine signaling via gamma-aminobutyric acid (GABA) and GABA(A) receptors (GABA(A)Rs) has been documented in rodent islets. Here we have studied the importance of GABAergic signaling in human pancreatic islets. Expression of GABA(A)Rs in islet cells was investigated by quantitative PCR, immunohistochemistry, and patch-clamp experiments. Hormone release was measured from intact islets. GABA release was monitored by whole-cell patch-clamp measurements after adenoviral expression of alpha(1)beta(1) GABA(A)R subunits. The subcellular localization of GABA was explored by electron microscopy. The effects of GABA on electrical activity were determined by perforated patch whole-cell recordings. PCR analysis detected relatively high levels of the mRNAs encoding GABA(A)R alpha(2), beta(3,) gamma(2), and pi subunits in human islets. Patch-clamp experiments revealed expression of GABA(A)R Cl(-) channels in 52% of beta-cells (current density 9 pA/pF), 91% of delta-cells (current density 148 pA/pF), and 6% of alpha-cells (current density 2 pA/pF). Expression of GABA(A)R subunits in islet cells was confirmed by immunohistochemistry. beta-Cells secreted GABA both by glucose-dependent exocytosis of insulin-containing granules and by a glucose-independent mechanism. The GABA(A)R antagonist SR95531 inhibited insulin secretion elicited by 6 mmol/l glucose. Application of GABA depolarized beta-cells and stimulated action potential firing in beta-cells exposed to glucose. Signaling via GABA and GABA(A)R constitutes an autocrine positive feedback loop in human beta-cells. The presence of GABA(A)R in non-beta-cells suggests that GABA may also be involved in the regulation of somatostatin and glucagon secretion.


SM22 a Plasma Biomarker for Human Transmural Intestinal Ischemia.
To evaluate the diagnostic potential of smooth muscle protein of 22 kDa (SM22) as plasma biomarker for the detection of transmural intestinal ischemia. Acute mesenteric ischemia is an abdominal emergency requiring rapid diagnosis and treatment. Especially, detection of transmural damage is imperative because it mandates emergency surgery. Since early clinical and radiological signs are nonspecific, there is an urgent need for accurate biomarkers. SM22 is a potential marker for transmural damage because of its abundant expression in intestinal smooth muscles. SM22 concentrations were measured using a newly built enzyme-linked immunosorbent assay. SM22 release was assessed in plasma and intestinal tissue of rats subjected to intestinal ischemia. Blood and tissue were sampled at baseline and followed up to 24 hours of ischemia. Next, organ-specific SM22 arteriovenous concentration differences were studied in both rats and patients. Finally, plasma from patients with intestinal ischemia, other acute abdominal complaints, and healthy volunteers were tested for SM22. SM22 concentrations were significantly elevated in rats from 4 hours of ischemia onwards. Furthermore, SM22 plasma concentrations closely paralleled the histological increasing degree of intestinal smooth muscle damage. Arteriovenous calculations showed that SM22 was specifically released by the intestines and renally cleared. First data of SM22 release in man demonstrated that patients with transmural intestinal ischemia had significantly higher plasma SM22 levels than patients with only ischemic mucosal injury, other acute abdominal diseases, or healthy controls. This study shows that SM22 is released into the circulation upon severe ischemia of the intestinal muscle layers. Plasma levels of SM22 are potentially useful for the detection of transmural intestinal damage.


Long-term survival outcomes of reduced-intensity allogeneic or autologous transplantation in relapsed grade 3 follicular lymphoma.
Grade 3 follicular lymphoma (FL) has aggressive clinical behavior. To evaluate the optimal first transplantation approach in relapsed/refractory grade 3 FL patients, we compared the long-term outcomes after allogeneic (allo-) vs autologous hematopoietic cell transplantation (auto-HCT) in the rituximab era. A total of 197 patients undergoing first reduced-intensity conditioning (RIC) allo-HCT or first auto-HCT during 2000-2012 were included. Rituximab-naive patients were excluded. Allo-HCT recipients were younger, more heavily pretreated and had a longer interval between diagnosis and HCT. The 5-year probabilities of non-relapse mortality (NRM), relapse/progression, PFS and overall survival (OS) for auto-HCT vs allo-HCT groups were 4% vs 27% (P<0.001), 61% vs 20% (P<0.001), 36% vs 51% (P=0.07) and 59% vs 54% (P=0.7), respectively. On multivariate analysis, auto-HCT was associated with reduced risk of NRM (relative risk (RR)=0.20; P=0.001). Within the first 11 months post HCT, auto- and allo-HCT had similar risks of relapse/progression and PFS. Beyond 11 months, auto-HCT was associated with higher risk of relapse/progression (RR=21.3; P=0.003) and inferior PFS (RR=3.2; P=0.005). In the first 24 months post HCT, auto-HCT was associated with improved OS (RR=0.42; P=0.005), but in long-time survivors (beyond 24 months) it was associated with inferior OS (RR=3.6; P=0.04). RIC allo-HCT as the first transplant approach can provide improved PFS and OS, in long-term survivors.


A SARM1-mitochondrial feedback loop drives neuropathogenesis in a Charcot-Marie-Tooth disease type 2A rat model.
Charcot-Marie-Tooth disease type 2A (CMT2A) is an axonal neuropathy caused by mutations in the mitofusin 2 (MFN2) gene. MFN2 mutations result in profound mitochondrial abnormalities, but the mechanism underlying the axonal pathology is unknown. Sterile α and Toll/IL-1 receptor motif-containing 1 (SARM1), the central executioner of axon degeneration, can induce neuropathy and is activated by dysfunctional mitochondria. We tested the role of SARM1 in a rat model carrying a dominant CMT2A mutation (Mfn2H361Y) that exhibits progressive dying-back axonal degeneration, neuromuscular junction (NMJ) abnormalities, muscle atrophy, and mitochondrial abnormalities - all hallmarks of the human disease. We generated Sarm1-KO (Sarm1-/-) and Mfn2H361Y Sarm1 double-mutant rats and found that deletion of Sarm1 rescued axonal, synaptic, muscle, and functional phenotypes, demonstrating that SARM1 was responsible for much of the neuropathology in this model. Despite the presence of mutant MFN2 protein in these double-mutant rats, loss of SARM1 also dramatically suppressed many mitochondrial defects, including the number, size, and cristae density defects of synaptic mitochondria. This surprising finding indicates that dysfunctional mitochondria activated SARM1 and that activated SARM1 fed back on mitochondria to exacerbate the mitochondrial pathology. As such, this work identifies SARM1 inhibition as a therapeutic candidate for the treatment of CMT2A and other neurodegenerative diseases with prominent mitochondrial pathology.


Dark matter in the genome: evidence of widespread transcription detected by microarray tiling experiments.
Microarrays provide the opportunity to measure transcription from regions of the genome without bias towards the location of known genes. This technology thus offers an important source of genomic sequence annotation that is complementary to cDNA sequencing and computational gene-finding methods. Recent "tiling" microarray experiments that assay transcription at regular intervals throughout the genome have shown evidence of large amounts of transcription outside the boundaries of known genes. This transcription is observed in polyadenylated RNA samples and appears to be derived from intergenic regions, from introns of known genes and from sequences antisense to known transcripts. In this article, we discuss different explanations for this phenomenon.


Domain size engineering in tetragonal Pb(In(1∕2)Nb(1∕2))O(3)-Pb(Mg(1∕3)Nb(2∕3))O(3)-PbTiO(3) crystals.
The effect of domain size on the dielectric and piezoelectric properties of [111]-oriented tetragonal Pb(In(1∕2)Nb(1∕2))O(3)-Pb(Mg(1∕3)Nb(2∕3))O(3)-PbTiO(3) crystals was investigated. The dielectric permittivity (ɛ(33 ) (T)∕ɛ(0)) and piezoelectric coefficient (d(33)) were found to be on the order of 13 800 and 1630 pC∕N, respectively, for samples with domain size of ∼500 nm, a 3-fold increase to crystals with domain size of ∼50 μm. Rayleigh analysis revealed that the extrinsic contribution to the piezoelectric response increased from ∼8% to 30% with decreasing domain size, due to the increased domain wall density and associated irreversible domain wall motion. The enhanced properties were thought to relate to the fine domain structures, however, showing a poor electric field and temperature stabilities with domain size of 500 nm. Of particular significance is that samples with domain size being on the order of 5 μm exhibit field and temperature stabilities, with yet high piezoelectric properties, make it potential for transducer applications.


Tissue geometry patterns epithelial-mesenchymal transition via intercellular mechanotransduction.
Epithelial-mesenchymal transition (EMT) is a phenotypic change in which epithelial cells detach from their neighbors and become motile. Whereas soluble signals such as growth factors and cytokines are responsible for stimulating EMT, here we show that gradients of mechanical stress define the spatial locations at which EMT occurs. When treated with transforming growth factor (TGF)-beta, cells at the corners and edges of square mammary epithelial sheets expressed EMT markers, whereas those in the center did not. Changing the shape of the epithelial sheet altered the spatial pattern of EMT. Traction force microscopy and finite element modeling demonstrated that EMT-permissive regions experienced the highest mechanical stress. Myocardin-related transcription factor (MRTF)-A was localized to the nuclei of cells located in high-stress regions, and inhibiting cytoskeletal tension or MRTF-A expression abrogated the spatial patterning of EMT. These data suggest a causal role for tissue geometry and endogenous mechanical stresses in the spatial patterning of EMT.


Speciation Rate Is Independent of the Rate of Evolution of Morphological Size, Shape, and Absolute Morphological Specialization in a Large Clade of Birds.
Whether ecological differences between species evolve in parallel with lineage diversification is a fundamental issue in evolutionary biology. These processes might be connected if conditions that favor the proliferation of species, such as release from competitors, facilitate the evolution of novel ecological relationships. Despite this, phylogenetic studies do not consistently identify such a connection. Conversely, if higher diversity caused species to become increasingly specialized ecologically, then lineage diversification might become dissociated from ecological diversification. In this analysis, we ask whether the rate of lineage diversification in a large clade of birds is correlated with morphological specialization and with rates of morphological evolution. We find that morphological variation is related to species richness within clades but that rates of morphological evolution are decoupled from the rate of lineage diversification. Additionally, morphological specialization within lineages is independent of the rate at which lineages diversify, with the results apparently robust against false negative inference. This dissociation is likely a consequence of the major ecomorphological differences between avian clades arising early in their evolutionary history, with comparatively little variation added subsequently, while avian diversification has been driven predominantly by geographic isolation and sexual selection. Accordingly, biodiversity appears to be limited by the extent to which taxa can subdivide exploited regions of ecological space and not just overall ecological opportunity.


G-quadruplexes as sensing probes.
Guanine-rich sequences of DNA are able to create tetrastranded structures known as G-quadruplexes; they are formed by the stacking of planar G-quartets composed of four guanines paired by Hoogsteen hydrogen bonding. G-quadruplexes act as ligands for metal ions and aptamers for various molecules. Interestingly, the G-quadruplexes form a complex with anionic porphyrin hemin and exhibit peroxidase-like activity. This review focuses on overview of sensing techniques based on G-quadruplex complexes with anionic porphyrins for detection of various analytes, including metal ions such as K+, Ca2+, Ag+, Hg2+, Cu2+, Pb2+, Sr2+, organic molecules, nucleic acids, and proteins. Principles of G-quadruplex-based detection methods involve DNA conformational change caused by the presence of analyte which leads to a decrease or an increase in peroxidase activity, fluorescence, or electrochemical signal of the used probe. The advantages of various detection techniques are also discussed.


1,25-Dihydroxyvitamin D3 induces differentiation of a retinoic acid-resistant acute promyelocytic leukemia cell line (UF-1) associated with expression of p21(WAF1/CIP1) and p27(KIP1).
Retinoic acid (RA) resistance is a serious problem for patients with acute promyelocytic leukemia (APL) who are receiving all-trans RA. However, the mechanisms and strategies to overcome RA resistance by APL cells are still unclear. The biologic effects of RA are mediated by two distinct families of transcriptional factors: RA receptors (RARs) and retinoid X receptors (RXRs). RXRs heterodimerize with 1, 25-dihydroxyvitamin D3 [1,25(OH)2D3] receptor (VDR), enabling their efficient transcriptional activation. The cyclin-dependent kinase (cdk) inhibitor p21(WAF1/CIP1) has a vitamin D3-responsive element (VDRE) in its promoter, and 1,25(OH)2D3 enhances the expression of p21(WAF1/CIP1) and induces differentiation of selected myeloid leukemic cell lines. We have recently established a novel APL cell line (UF-1) with features of RA resistance. 1,25(OH)2D3 can induce growth inhibition and G1 arrest of UF-1 cells, resulting in differentiation of these cells toward granulocytes. This 1, 25(OH)2D3-induced G1 arrest is enhanced by all-trans RA. Also, 1, 25(OH)2D3 (10(-10) to 10(-7) mol/L) in combination with RA markedly inhibits cellular proliferation in a dose- and time-dependent manner. Associated with these findings, the levels of p21(WAF1/CIP1) and p27(KIP1) mRNA and protein increased in these cells. Northern blot analysis showed that p21(WAF1/CIP1) and p27(KIP1) mRNA and protein increased in these cells. Northern blot analysis showed that p21(WAF1/CIP1) and p27(KIP1) transcripts were induced after 6 hours' exposure to 1,25(OH)2D3 and then decreased to basal levels over 48 hours. Western blot experiments showed that p21(WAF1/CIP1) protein levels increased and became detectable after 12 hours of 1,25(OH)2D3 treatment and induction of p27(KIP1) protein was much more gradual and sustained in UF-1 cells. Interestingly, the combination of 1, 25(OH)2D3 and RA markedly enhanced the levels of p27(KIP1) transcript and protein as compared with levels induced by 1, 25(OH)2D3 alone. In addition, exogenous p27(KIP1) expression can enhance the level of CD11b antigen in myeloid leukemic cells. In contrast, RA alone can induce G1 arrest of UF-1 cells; however, it did not result in an increase of p21(WAF1/CIP1) and p27(KIP1) transcript and protein expression in RA-resistant cells. Taken together, we conclude that 1,25(OH)2D3 induces increased expression of cdk inhibitors, which mediates a G1 arrest, and this may be associated with differentiation of RA-resistant UF-1 cells toward mature granulocytes.


c-myc in the hematopoietic lineage is crucial for its angiogenic function in the mouse embryo.
The c-myc proto-oncogene, which is crucial for the progression of many human cancers, has been implicated in key cellular processes in diverse cell types, including endothelial cells that line the blood vessels and are critical for angiogenesis. The de novo differentiation of endothelial cells is known as vasculogenesis, whereas the growth of new blood vessels from pre-existing vessels is known as angiogenesis. To ascertain the function of c-myc in vascular development, we deleted c-myc in selected cell lineages. Embryos lacking c-myc in endothelial and hematopoietic lineages phenocopied those lacking c-myc in the entire embryo proper. At embryonic day (E) 10.5, both mutant embryos were grossly normal, had initiated primitive hematopoiesis, and both survived until E11.5-12.5, longer than the complete null. However, they progressively developed defective hematopoiesis and angiogenesis. The majority of embryos lacking c-myc specifically in hematopoietic cells phenocopied those lacking c-myc in endothelial and hematopoietic lineages, with impaired definitive hematopoiesis as well as angiogenic remodeling. c-myc is required for embryonic hematopoietic stem cell differentiation, through a cell-autonomous mechanism. Surprisingly, c-myc is not required for vasculogenesis in the embryo. c-myc deletion in endothelial cells does not abrogate endothelial proliferation, survival, migration or capillary formation. Embryos lacking c-myc in a majority of endothelial cells can survive beyond E12.5. Our findings reveal that hematopoiesis is a major function of c-myc in embryos and support the notion that c-myc functions in selected cell lineages rather than in a ubiquitous manner in mammalian development.


Ranking of parameters of pain hypersensitivity according to their discriminative ability in chronic low back pain.
Low back pain is associated with plasticity changes and central hypersensitivity in a subset of patients. We performed a case-control study to explore the discriminative ability of different quantitative sensory tests in distinguishing between 40 cases with chronic low back pain and 300 pain-free controls, and to rank these tests according to the extent of their association with chronic pain. Gender, age, height, weight, body mass index, and psychological measures were recorded as potential confounders. We used 26 quantitative sensory tests, including different modalities of pressure, heat, cold, and electrical stimulation. As measures of discrimination, we estimated receiver operating characteristics (ROC) and likelihood ratios. Six tests seemed useful (in order of their discriminative ability): (1) pressure pain detection threshold at the site of most severe pain (fitted area under the ROC, 0.87), (2) single electrical stimulation pain detection threshold (0.87), (3) single electrical stimulation reflex threshold (0.83), (4) pressure pain tolerance threshold at the site of most severe pain (0.81), (5) pressure pain detection threshold at suprascapular region (0.80), and (6) temporal summation pain threshold (0.80). Pressure and electrical pain modalities seemed most promising and may be used for diagnosis of pain hypersensitivity and potentially for identifying individuals at risk of developing chronic low back pain over time.


Enhancement of hydroxyl radical generation in the Fenton reaction by alpha-hydroxy acid.
The effect of various organic acids on hydroxyl radical (.OH) generation in the Fenton reaction were examined by the ESR spin trapping technique, where 5,5-dimethyl-1-pyroline-N-nitroxide (DMPO) and alpha-phenyl-tert-butyl nitrone (PBN) were used as the spin trapping reagents. alpha-Hydroxy acids such as lactic acid, glycolic acid and 2-hydroxy isobutyric acid were found to markedly enhance .OH generation in the reaction. In contrast, beta-hydroxy acid, alpha-keto acid, esters of alpha-hydroxy acids, aldehydes and other straight chain organic acids had no such enhancing activity. alpha-Amino acids had also no enhancing effect. The results suggest that the alpha-hydroxy acid moiety is prerequisite for the enhancement of .OH generation in the Fenton reaction. Superoxide dismutase did not inhibit the enhancing effect of alpha-hydroxy acids whereas catalase completely inhibited the .OH generation. Thus, alpha-hydroxy acids directly enhanced the .OH generation via the Fenton reaction but not the Haber-Weiss reaction. Possible role of lactic acid manipulating .OH generation is discussed in relation to the ischemia-reperfusion cell damage.


Young children's perceptions of fire-safety messages: do framing and parental mediation matter?
Media can deliver health and safety messages promoting child health and injury prevention. This study examined the effects of message framing and parental mediation on children's perceptions of fire-safety messages. Using a 2 × 3 randomized experimental design, this study considered both message framing (gain or loss) and parental mediation (no mediation/control, unscripted, or scripted) with 320 children who were 4 and 5 years of age. Children saw two messages (burn and smoke inhalation) embedded in a cartoon. Afterward, researchers assessed children's recall, understanding, and perceptions of self-efficacy and social norms. Children were more likely to recall the safety messages if they were older (burn: adjusted odds ration [AOR] = 2.74 and smoke: AOR = 2.58), and could recall the smoke inhalation message if they had unscripted mediation (AOR = 3.16). Message understanding was poor, with only about 50% of children choosing a correct behavior in a similar scenario. For the burn message, correct understanding was associated with gain-framing and scripted mediation (AOR = 3.22 and 5.77, respectively). Only the scripted mediation group was significantly associated with an increase in perceived social norms (burn: coefficient =.37 and smoke: coefficient =.55; P <.001. Gain-framing was associated with increased odds of self-efficacy for both behaviors (burn: AOR = 1.77 and smoke: AOR = 1.77). Messages that show positive outcomes combined with scripted parental mediation appear most effective in communicating safety behaviors, but the overall effectiveness of video-based messages to teach children safety behaviors needs to be enhanced.


TRICALS: creating a highway toward a cure.
A change in our current approach toward drug development is required to improve the likelihood of finding effective treatment for patients with amyotrophic lateral sclerosis (ALS). The aim of the Treatment Research Initiative to Cure ALS (TRICALS) is to extend the collective effort with industry and consolidate drug development paths. TRICALS has begun a series of meetings on how to best move the field forward collaboratively, thereby addressing five major topics in ALS clinical trials: (1) preclinical research, (2) biomarker development, (3) eligibility criteria, (4) efficacy endpoints and (5) innovative trial design. There is an appetite for ongoing discussions of these major topics in clinical trials between representatives from academia, patient advocacy groups, industry partners and funding bodies. Industry is open to fundamentally change drug development for ALS and shorten the time to effective therapy for patients by implementing promising innovations in biomarker development, trial design, and patient selection. There is however, a pressing need from all stakeholders for regulatory discussions and amendments of current guidelines to successfully adopt innovation in future clinical development lines.


Cyclooctatetraenyl calcium and strontium amido complexes.
This paper reports the synthesis and structural characterisation of a series of cyclooctatetraenyl (C8H8) complexes of calcium and strontium, most of them containing the bis(trimethylsilyl)amido ligand. Mixing MI2 (M = Ca, Sr), KN(SiMe3)2 and K2(C8H8) in thf in a 2 : 2 : 1 ratio gave the inverse sandwich amido complexes [[{(Me3Si)2N}M(thf)x]2(μ-C8H8)] (M = Ca, x = 1; M = Sr, x = 2) (1-Ca, 1-Sr) in fair to good yields. From a 1 : 1 : 1 ratio, the mixed potassium calcium complex [[{(Me3Si)2N}Ca(thf)](μ-C8H8)K] (2) was obtained in good yield. The reaction of 2 with a 1 : 1 mixture of CaI2 and KN(SiMe3)2 yielded 1-Ca confirming that the reaction could be carried out stepwise. Attempts at making heterobimetallic calcium strontium amido complexes from the reaction of 2 with Sr{N(SiMe3)2}2 led to redistribution reactions which afforded the potassium strontium complex [[K{(Me3Si)2N}2Sr]2(μ-C8H8)] (3) among other species. Complex 3 was more conveniently synthesised in fair yield starting from a 2 : 4 : 1 molar mixture of SrI2, KN(SiMe3)2 and K2(C8H8), respectively. Treatment of 2 with PhC[triple bond, length as m-dash]CH in benzene yielded an insoluble complex tentatively formulated as the polymeric complex [CaK(CCPh)(C8H8)(thf)x]n (4). Upon dissolution in thf, 4 underwent a redistribution reaction yielding the X-ray characterised [{Ca(μ-C8H8)2}K2(thf)2] (5) and a homoleptic calcium alkynyl complex [Ca(CCPh)2(thf)x]. Several compounds were characterised by X-ray diffraction as discrete or polymeric structures where CaCπ interactions were identified in some cases. Despite identified drawbacks, such as the lability of the coordinated thf ligands and facile redistribution reactions, this study represents the first synthetic approach to cyclooctatetraenyl heavy alkaline-earth metal amido complexes.


Exploring the experiences of living with Lewy body dementia: An integrative review.
Lewy body dementia is a common neurodegenerative dementia with unique challenges in managing day-to-day life. A more in-depth multifaceted picture of the Lewy body dementia lived experience will enable identification of best practice and future research direction. The review aim was to explore experiences of people living with Lewy body dementia and their family carers. Integrative review method informed by Whittemore and Knafl, supported by the information retrieval framework PALETTE. A convergent integrated approach enabled synthesis of key findings and theme identification. Medline, CINAHL, PsycINFO, AMED, and ALOIS databases were systematically searched to find studies published between 1995 and 2020. Twenty-six articles from twenty studies were synthesized (from 1583 retrieved). Quality and relevance were appraised using the Mixed-Methods Appraisal Tool and Gough's 'Weight of Evidence' framework. Data management was supported by ATLAS.ti 8 and COVIDENCE software. Four themes were identified: living with an uncertain diagnosis and prognosis; fear of the now - worry for the future; living with behavioural and psychiatric symptoms; and maintaining a social and emotional life. People reported difficulty finding information and support around diagnosis, disease progression and managing complex symptoms. The result is increased caregiver burden, grief and stress and reduced quality of life. Delayed diagnosis and complex symptom burden means people are not getting the timely support and information they need to live and die well. Current evidence is largely quantitative, with a focus on family caregiver burden and unmet need. The challenge remains in how to capture a more holistic picture of the lived experience for people living with Lewy body dementia and those who care for them. This review highlighted current knowledge and identified gaps in exploring the lived experience for people with Lewy body dementia and their families.


Global expansion and redistribution of Aedes-borne virus transmission risk with climate change.
Forecasting the impacts of climate change on Aedes-borne viruses-especially dengue, chikungunya, and Zika-is a key component of public health preparedness. We apply an empirically parameterized model of viral transmission by the vectors Aedes aegypti and Ae. albopictus, as a function of temperature, to predict cumulative monthly global transmission risk in current climates, and compare them with projected risk in 2050 and 2080 based on general circulation models (GCMs). Our results show that if mosquito range shifts track optimal temperature ranges for transmission (21.3-34.0°C for Ae. aegypti; 19.9-29.4°C for Ae. albopictus), we can expect poleward shifts in Aedes-borne virus distributions. However, the differing thermal niches of the two vectors produce different patterns of shifts under climate change. More severe climate change scenarios produce larger population exposures to transmission by Ae. aegypti, but not by Ae. albopictus in the most extreme cases. Climate-driven risk of transmission from both mosquitoes will increase substantially, even in the short term, for most of Europe. In contrast, significant reductions in climate suitability are expected for Ae. albopictus, most noticeably in southeast Asia and west Africa. Within the next century, nearly a billion people are threatened with new exposure to virus transmission by both Aedes spp. in the worst-case scenario. As major net losses in year-round transmission risk are predicted for Ae. albopictus, we project a global shift towards more seasonal risk across regions. Many other complicating factors (like mosquito range limits and viral evolution) exist, but overall our results indicate that while climate change will lead to increased net and new exposures to Aedes-borne viruses, the most extreme increases in Ae. albopictus transmission are predicted to occur at intermediate climate change scenarios.


Endoscopic ablation of intestinal metaplasia containing high-grade dysplasia in esophagectomy patients using a balloon-based ablation system.
This study aimed to determine the optimal treatment parameters for the ablation of intestinal metaplasia (IM) containing high-grade dysplasia (HGD) using a balloon-based ablation system for patients undergoing esophagectomy. Immediately before esophagectomy, patients underwent ablation of circumferential segments of the esophagus containing IM-HGD using the HALO360 system. The treatment settings were randomized to 10, 12, or 14 J/cm2 for two, three, or four applications. After esophagectomy, multiple sections from ablation zones were microscopically evaluated. Histologic end points included maximum ablation depth (histologic layer) and complete ablation of all IM-HGD (yes/no). Eight men with a mean age of 57 years (range, 45-71 years) were treated, and 10 treatment zones were created. There were no device-related adverse events. At resection, there was no evidence of a transmural thermal effect. Grossly, ablation zones were clearly demarcated sections of ablated epithelium. The maximum ablation depth was the lamina propria or muscularis mucosae. The highest energy (14 J/cm2, 4 applications) incurred edema in the superficial submucosa, but no submucosa ablation. Complete ablation of IM and HGD occurred in 9 of 10 ablation zones (90%), defined as complete removal of the epithelium with only small foci of "ghost cells" representing nonviable, ablated IM-HGD and demonstrating loss of nuclei and cytoarchitectural derangement. One focal area of viable IM-HGD remained at the margin of one ablation zone (12 J/cm2, 2 applications) because of incomplete overlap. Complete ablation of IM-HGD without ablation of submucosa is possible using the HALO360 system. Ablation depth is dose related and limited to the muscularis mucosae. In one patient, small residual foci of IM-HGD at the edge of the ablation zone were attributable to incomplete overlap, which can be avoided. This study, together with nonesophagectomy IM-HGD trials currently underway, will identify the optimal treatment parameters for IM-HGD patients who would otherwise undergo esophagectomy or photodynamic therapy.


Role of beta blockers following percutaneous coronary intervention for acute coronary syndrome.
There is a paucity of evidence supporting routine beta blocker (BB) use in patients undergoing percutaneous coronary intervention (PCI) for acute coronary syndrome (ACS). The aim of this study was to evaluate BB use post PCI and its association with mortality. Furthermore, the study aimed to evaluate the association between BB and mortality in the subgroups of patients with left ventricular ejection fraction (LVEF) <35%, LVEF 35%-50% and LVEF >50%. Using a large PCI registry, data from patients with ACS between January 2005 and June 2017 who were alive at 30 days were analysed. Those patients taking BB at 30 days were compared with those who were not taking BB. The primary outcome was all-cause mortality. The mean follow-up was 5.3±3.5 years. Of the 17 562 patients, 83.3% were on BB. Mortality was lower in the BB group (13.1% vs 19.5%, p=0.0001). Multivariable Cox proportional hazards model showed that BB use was associated with lower overall mortality (adjusted HR 0.87, 95% CI 0.78 to 0.97, p=0.014). In the subgroup analysis, BB use was associated with reduced mortality in LVEF <35% (adjusted HR 0.63, 95% CI 0.44 to 0.91, p=0.013), LVEF 35%-50% (adjusted HR 0.80, 95% CI 0.68 to 0.95, p=0.01), but not LVEF >50% (adjusted HR 1.03, 95% CI 0.87 to 1.21, p=0.74). BB use remains high and is associated with reduced mortality. This reduction in mortality is primarily seen in those with reduced ejection fraction, but not in those with preserved ejection fraction.


Cost-effectiveness of the Mental Health and Development model for schizophrenia-spectrum and bipolar disorders in rural Kenya.
The treatment gap for serious mental disorders across low-income countries is estimated to be 89%. The model for Mental Health and Development (MHD) offers community-based care for people with mental disorders in 11 low- and middle-income countries. In Kenya, using a pre-post design, 117 consecutively enrolled participants with schizophrenia-spectrum and bipolar disorders were followed-up at 10 and 20 months. Comparison outcomes were drawn from the literature. Costs were analysed from societal and health system perspectives. From the societal perspective, MHD cost Int$ 594 per person in the first year and Int$ 876 over 2 years. The cost per healthy day gained was Int$ 7.96 in the first year and Int$ 1.03 over 2 years - less than the agricultural minimum wage. The cost per disability-adjusted life year averted over 2 years was Int$ 13.1 and Int$ 727 from the societal and health system perspectives, respectively, on par with antiretrovirals for HIV. MHD achieved increasing returns over time. The model appears cost-effective and equitable, especially over 2 years. Its affordability relies on multi-sectoral participation nationally and internationally.