Incorrect use of the student t test in randomized trials of bilateral hip and knee arthroplasty patients.
The statistical analysis of a study's results is critical to its interpretation. Often confused, 2 types of t tests exist for dependent or independent samples. We reviewed randomized controlled trials that included patients undergoing bilateral total hip or knee arthroplasty (dependent samples) that used the t test for analysis. Our inclusion criteria resulted in 40 studies for review, 10 of which combined both unilateral and bilateral patients. Of the 30 studies that compared a pure sample of only bilateral patients, 18 (60%) incorrectly used the independent t test. Of the 10 studies with mixed samples, 8 (80%) used the independent t test. The extent to which the incorrect use of this statistical test led to misleading conclusions is uncertain; however, our findings highlight the misuse of statistical tests in the arthroplasty literature.


Ethylene regulates fast apoplastic acidification and expansin A transcription during submergence-induced petiole elongation in Rumex palustris.
The semi-aquatic dicot Rumex palustris responds to complete submergence by enhanced elongation of young petioles. This elongation of petiole cells brings leaf blades above the water surface, thus reinstating gas exchange with the atmosphere and increasing survival in flood-prone environments. We already know that an enhanced internal level of the gaseous hormone ethylene is the primary signal for underwater escape in R. palustris. Further downstream, concentration changes in abscisic acid (ABA), gibberellin (GA) and auxin are required to gain fast cell elongation under water. A prerequisite for cell elongation in general is cell wall loosening mediated by proteins such as expansins. Expansin genes might, therefore, be important target genes in submergence-induced and plant hormone-mediated petiole elongation. To test this hypothesis we have studied the identity, kinetics and regulation of expansin A mRNA abundance and protein activity, as well as examined pH changes in cell walls associated with this adaptive growth. We found a novel role of ethylene in triggering two processes affecting cell wall loosening during submergence-induced petiole elongation. First, ethylene was shown to promote fast net H(+) extrusion, leading to apoplastic acidification. Secondly, ethylene upregulates one expansin A gene (RpEXPA1), as measured with real-time RT-PCR, out of a group of 13 R. palustris expansin A genes tested. Furthermore, a significant accumulation of expansin proteins belonging to the same size class as RpEXPA1, as well as a strong increase in expansin activity, were apparent within 4-6 h of submergence. Regulation of RpEXPA1 transcript levels depends on ethylene action and not on GA and ABA, demonstrating that ethylene evokes at least three, parallel operating pathways that, when integrated at the whole petiole level, lead to coordinated underwater elongation. The first pathway involves ethylene-modulated changes in ABA and GA, these acting on as yet unknown downstream components, whereas the second and third routes encompass ethylene-induced apoplastic acidification and ethylene-induced RpEXPA1 upregulation.


The Patient Care Paradox: An Interprofessional Qualitative Study of Agitated Patient Care in the Emergency Department.
The emergency department (ED) has been recognized as a high-risk environment for workplace violence. Acutely agitated patients who perpetrate violence against healthcare workers represent a complex care challenge in the ED. Recommendations to improve safety are often based on expert opinion rather than empirical data. In this study we aim to describe the lived experience of staff members caring for this population to provide a broad perspective of ED patient violence. The findings of this study will contribute to the development of a comprehensive framework for ED agitated patient care that will guide safety interventions. We conducted uniprofessional focus groups and individual interviews using a phenomenologic approach with emergency medicine resident physicians, ED staff nurses, patient care technicians, and hospital police officers at an urban hospital in New York City. Audio recordings were transcribed and coded for thematic analysis using the constant comparison method. We reached theoretical saturation with 31 interprofessional participants. Three broad themes emerged from our analysis: 1) ED healthcare workers provide high-quality care to a marginalized patient population that concurrently poses safety threats, creating a patient care paradox; 2) teamwork is critical to safely managing this population, but hierarchy and professional silos hinder coordinated care between healthcare professionals; and 3) environmental challenges and systems issues both in and outside the ED exacerbate threats to safety. The experience of ED staff members while caring for agitated patients is complex and multidimensional. We identified issues that coalesced into four tiers of healthcare delivery at the individual, team, environment, and system levels. Future research is needed to determine applicability of our findings across institutions to build a comprehensive framework for ED agitated patient care.


Human-specific duplication and mosaic transcripts: the recent paralogous structure of chromosome 22.
In recent decades, comparative chromosomal banding, chromosome painting, and gene-order studies have shown strong conservation of gross chromosome structure and gene order in mammals. However, findings from the human genome sequence suggest an unprecedented degree of recent (<35 million years ago) segmental duplication. This dynamism of segmental duplications has important implications in disease and evolution. Here we present a chromosome-wide view of the structure and evolution of the most highly homologous duplications (> or = 1 kb and > or = 90%) on chromosome 22. Overall, 10.8% (3.7/33.8 Mb) of chromosome 22 is duplicated, with an average sequence identity of 95.4%. To organize the duplications into tractable units, intron-exon structure and well-defined duplication boundaries were used to define 78 duplicated modules (minimally shared evolutionary segments) with 157 copies on chromosome 22. Analysis of these modules provides evidence for the creation or modification of 11 novel transcripts. Comparative FISH analyses of human, chimpanzee, gorilla, orangutan, and macaque reveal qualitative and quantitative differences in the distribution of these duplications--consistent with their recent origin. Several duplications appear to be human specific, including a approximately 400-kb duplication (99.4%-99.8% sequence identity) that transposed from chromosome 14 to the most proximal pericentromeric region of chromosome 22. Experimental and in silico data further support a pericentromeric gradient of duplications where the most recent duplications transpose adjacent to the centromere. Taken together, these data suggest that segmental duplications have been an ongoing process of primate genome evolution, contributing to recent gene innovation and the dynamic transformation of genome architecture within and among closely related species.


Global Mortality Impact of the 1957-1959 Influenza Pandemic.
Quantitative estimates of the global burden of the 1957 influenza pandemic are lacking. Here we fill this gap by modeling historical mortality statistics. We used annual rates of age- and cause-specific deaths to estimate pandemic-related mortality in excess of background levels in 39 countries in Europe, the Asia-Pacific region, and the Americas. We modeled the relationship between excess mortality and development indicators to extrapolate the global burden of the pandemic. The pandemic-associated excess respiratory mortality rate was 1.9/10,000 population (95% confidence interval [CI], 1.2-2.6 cases/10,000 population) on average during 1957-1959. Excess mortality rates varied 70-fold across countries; Europe and Latin America experienced the lowest and highest rates, respectively. Excess mortality was delayed by 1-2 years in 18 countries (46%). Increases in the mortality rate relative to baseline were greatest in school-aged children and young adults, with no evidence that elderly population was spared from excess mortality. Development indicators were moderate predictors of excess mortality, explaining 35%-77% of the variance. Overall, we attribute 1.1 million excess deaths (95% CI, .7 million-1.5 million excess deaths) globally to the 1957-1959 pandemic. The global mortality rate of the 1957-1959 influenza pandemic was moderate relative to that of the 1918 pandemic but was approximately 10-fold greater than that of the 2009 pandemic. The impact of the pandemic on mortality was delayed in several countries, pointing to a window of opportunity for vaccination in a future pandemic.


Freezing of Solute-Laden Aqueous Solutions: Kinetics of Crystallization and Heat- and Mass-Transfer-Limited Model.
Following an earlier study, we reexamined the latent heat of fusion during freezing at 5 K/min of twelve different pre-nucleated solute-laden aqueous solutions using a Differential Scanning Calorimeter (DSC) and correlated it with the amount of initially dissolved solids or solutes in the solution. In general, a decrease in DSC-measured heat release (in comparison to that of pure water, 335 mJ/mg) was observed with an increasing fraction of dissolved solids or solutes, as observed in the earlier study. In addition, the kinetics of ice crystallization was also obtained in three representative biological media by performing additional experiments at 1, 5 and 20 K/min. A model of ice crystallization based on the phase diagram of a water-NaCl binary solution and a modified Avrami-like model of kinetics was then developed and fit to the experimental data. Concurrently, a heat and mass transfer model of the freezing of a salt solution in a small container is also presented to account for the effect of the cooling rate as well as the solute concentration on the measured latent of freezing. This diffusion-based model of heat and mass transfer was non-dimensionalized, solved using a numerical scheme and compared with experimental results. The simulation results show that the heat and mass transfer model can predict (± 10%) the experimental results.


Structure-activity relationship studies on isoindoline inhibitors of dipeptidyl peptidases 8 and 9 (DPP8, DPP9): is DPP8-selectivity an attainable goal?
This work represents the first directed study to identify modification points in the topology of a representative DPP8/9-inhibitor, capable of rendering selectivity for DPP8 over DPP9. The availability of a DPP8-selective compound would be highly instrumental for studying and untwining the biological roles of DPP8 and DPP9 and for the disambiguation of biological effects of nonselective DPP-inhibitors that have mainly been ascribed to blocking of DPPIV's action. The cell-permeable DPP8/9-inhibitor 7 was selected as a lead and dissected into several substructures that were modified separately for evaluating their potential to contribute to selectivity. The obtained results, together with earlier work from our group, clearly narrow down the most probable DPP8-selectivity imparting modification points in DPP8/9 inhibitors to parts of space that are topologically equivalent to the piperazine ring system in 7. This information can be considered of high value for future design of compounds with maximal DPP8 selectivity.


Epidemiology of Inflammatory Bowel Diseases in Nepal.
Introduction Inflammatory bowel diseases (IBD) comprise ulcerative colitis (UC) and Crohn's disease (CD). These are diseases of the gastrointestinal tract without a clear etiology but have strong relationships with underlying factors like genetic susceptibility, environmental factors, and intestinal bacteria. In the east, inflammatory bowel diseases predominantly affect the younger population and have an almost equal gender distribution. With urbanization and the adoption of the western lifestyle, the incidence and prevalence of IBD are increasing in Asia. In this study, we describe the epidemiology of IBD in Nepal. Methods This was an observational study conducted in nine endoscopy centers within Nepal. Two years of data of colonoscopies in these centers were collected retrospectively. IBD was diagnosed by endoscopic examination. The incidence of IBD was calculated as the number of patients with IBD per 1000 colonoscopies per year. The demographic profiles of the patients were also collected. Results A total of 7526 colonoscopies were done in nine centers within the two years study period. IBD was seen in 479 patients (6.3%). The incidence of UC was 23.7 per 1000 colonoscopies per year and the incidence of CD was 1.6 per 1000 colonoscopies per year. UC (87%) was more common than CD (13%). Both UC and CD were mostly seen in the 30 to 40 years age group. In patients with UC, the rectum was the most commonly affected site. Discussion IBD in Nepal affects young males in their thirties. Younger age of affliction with a chronic disease and lack of awareness regarding the symptoms and diagnostic modalities of IBD may result in a delayed diagnosis. The target population must be made aware of the presenting symptoms of IBD and a need for colonoscopic examination for diagnosis. There is also a need for creating a national IBD registry for Nepal.


Developmental plasticity and epigenetic mechanisms underpinning metabolic and cardiovascular diseases.
The importance of developmental factors in influencing the risk of later-life disease has a strong evidence base derived from multiple epidemiological, clinical and experimental studies in animals and humans. During early life, an organism is able to adjust its phenotypic development in response to environmental cues. Such developmentally plastic responses evolved as a fitness-maximizing strategy to cope with variable environments. There are now increasing data that these responses are, at least partially, underpinned by epigenetic mechanisms. A mismatch between the early and later-life environments may lead to inappropriate early life-course epigenomic changes that manifest in later life as increased vulnerability to disease. There is also growing evidence for the transgenerational transmission of epigenetic marks. This article reviews the evidence that susceptibility to metabolic and cardiovascular disease in humans is linked to changes in epigenetic marks induced by early-life environmental cues, and discusses the clinical, public health and therapeutic implications that arise.


Estimated macronutrient and fatty acid intakes from an East African Paleolithic diet.
Our genome adapts slowly to changing conditions of existence. Many diseases of civilisation result from mismatches between our Paleolithic genome and the rapidly changing environment, including our diet. The objective of the present study was to reconstruct multiple Paleolithic diets to estimate the ranges of nutrient intakes upon which humanity evolved. A database of, predominantly East African, plant and animal foods (meat/fish) was used to model multiple Paleolithic diets, using two pathophysiological constraints (i.e. protein < 35 energy % (en%) and linoleic acid (LA) >1.0 en%), at known hunter-gatherer plant/animal food intake ratios (range 70/30-30/70 en%/en%). We investigated selective and non-selective savannah, savannah/aquatic and aquatic hunter-gatherer/scavenger foraging strategies. We found (range of medians in en%) intakes of moderate-to-high protein (25-29), moderate-to-high fat (30-39) and moderate carbohydrates (39-40). The fatty acid composition was SFA (11.4-12.0), MUFA (5.6-18.5) and PUFA (8.6-15.2). The latter was high in α-linolenic acid (ALA) (3.7-4.7 en%), low in LA (2.3-3.6 en%), and high in long-chain PUFA (LCP; 4.75-25.8 g/d), LCP n-3 (2.26-17.0 g/d), LCP n-6 (2.54-8.84 g/d), ALA/LA ratio (1.12-1.64 g/g) and LCP n-3/LCP n-6 ratio (0.84-1.92 g/g). Consistent with the wide range of employed variables, nutrient intakes showed wide ranges. We conclude that compared with Western diets, Paleolithic diets contained consistently higher protein and LCP, and lower LA. These are likely to contribute to the known beneficial effects of Paleolithic-like diets, e.g. through increased satiety/satiation. Disparities between Paleolithic, contemporary and recommended intakes might be important factors underlying the aetiology of common Western diseases. Data on Paleolithic diets and lifestyle, rather than the investigation of single nutrients, might be useful for the rational design of clinical trials.


Pseudomonas aeruginosa small protease (PASP), a keratitis virulence factor.
The virulence contribution of Pseudomonas aeruginosa small protease (PASP) during experimental keratitis was studied by comparing a PASP-deficient mutant with its parent and rescue strains. The pasP gene of P. aeruginosa was replaced with the tetracycline resistance gene via allelic exchange. A plasmid carrying the pasP gene was introduced into the PASP-deficient mutant to construct a rescue strain. The PASP protein in the culture supernatants was determined by Western blot analysis. Corneal virulence was evaluated in rabbit and mouse keratitis models by slit lamp examination (SLE), bacterial enumeration, and/or histopathological analysis. Various host proteins and the rabbit tear film were analyzed for their susceptibility to PASP degradation. The PASP-deficient mutant produced a significantly lower mean SLE score when compared with the parent or rescue strain (P ≤ 0.03) at 29 hours postinfection (PI). All of the strains grew equally in the rabbit cornea (P = 0.971). Corneas infected with the PASP-deficient mutant showed moderate histopathology compared with those infected with the parent or rescue strain, which produced severe pathology inclusive of epithelial erosions, corneal edema, and neutrophil infiltration. In the mouse model, eyes inoculated with the PASP-deficient mutant had a significantly lower mean SLE score at 24 hours PI than the eyes inoculated with the parent or rescue strain (P ≤ 0.007). PASP was found to degrade complement C3, fibrinogen, antimicrobial peptide LL-37, and constituents of the tear film. PASP is a commonly secreted protease of P. aeruginosa that contributes significantly to the pathogenesis of keratitis.


Incorporating Sex as a Biological Variable into Clinical and Translational Research Training.
Incorporating sex as a biological variable (SABV) into basic and medical research requires a deliberate plan that weaves concepts of basic genetics, cellular and molecular physiology, and pharmacology into translational medicine. An R4 approach (Right content to the Right learner at the Right time with the Right modality) allows for content to be available in a variety of formats that reinforces the concepts at staged levels of integration. Weaving SABV throughout the varied formats of the R4 approach within the Clinical and Translational Research Training Programs, into the mentoring and training of scholars in NIH Building Interdisciplinary Careers in Women's Health (BIRCWH), and into the Career Enhancement Core of the Specialized Centers of Research Excellence (SCORE) on sex differences through curriculum, case-based approach and journal clubs, and workshops ensures that learners grasp its fundamental relevancy to their own research and beyond. In addition, the collaborative work among the BIRCWH and SCORE programs brings collective expertise from centers around the United States to individual programs through development of best practices and materials. These collective efforts assure that the next generation of basic, clinical, and translational scientists will bring the dimension of SABV into their research and clinical practice.


Management of elderly patients with troponin positive chest pain in a district general hospital.
The number of elderly patients that present with an acute coronary syndrome (ACS) is increasing, reflecting the growing number of people in the general population in this age group. The various guidelines do not generally specify a management strategy in this elderly group and the management is often at the discretion of the treating physician. We conducted an audit within our Cardiology Department to compare our practice of management of ACS in the elderly population based on the European Society of Cardiology guidelines. We conducted a retrospective analysis of the management of patients aged 80 and above that were admitted with troponin positive chest pain from 1(st) January to 31(st) December 2010. Patient information was primarily obtained from our computer data base system that includes blood results, ECHOs, diagnostic angiograms, discharge and clinic letters. If the information was inadequate we obtained patient files or contacted the relevant general practitioner. Octo-nonagenarians represented just over a third (35%) of all patients that were admitted with a troponin positive event during the study period. We noted a 10% mortality rate observed in our study population over a 12 month period. Atrial fibrillation was an incidental finding in 22% of patients. Nearly half of these patients (49%) were managed by the cardiologists. 68% of these patients underwent diagnostic coronary angiography, of which 32% went on to have percutaneous coronary intervention and 7% underwent surgical intervention. Majority (80%) of patients that underwent angioplasty had more than 1 stent and 74% of patients required more than one coronary vessel to be stented. The length of stay in hospital was double for patients who were under the care of the general medical teams rather than the cardiology team. This group also had a higher number of other comorbidities such as dementia, malignancy, a history of gastro intestinal bleeds and chronic renal impairment. Octo and nonagenarians represent a significant proportion of our ACS patients. They have high mortality, greater number of comorbidities, diseased coronary vessels and if intervention was undertaken required more than one stent. Therefore, octo-nonagenarians represent a very complex group of patients. Guidelines and risk stratification are of limited value in this group as clinical trial data is currently lacking. Quality of life and risk to benefit assessments are of paramount importance in this group.


Integrative genomics analysis identifies candidate drivers at 3q26-29 amplicon in squamous cell carcinoma of the lung.
Chromosome 3q26-29 is a critical region of genomic amplification in lung squamous cell carcinomas (SCC). Identification of candidate drivers in this region could help uncover new mechanisms in the pathogenesis and potentially new targets in SCC of the lung. We conducted a meta-analysis of seven independent datasets containing a total of 593 human primary SCC samples to identify consensus candidate drivers in 3q26-29 amplicon. Through integrating protein-protein interaction network information, we further filtered for candidates that may function together in a network. Computationally predicted candidates were validated using RNA interference (RNAi) knockdown and cell viability assays. Clinical relevance of the experimentally supported drivers was evaluated in an independent cohort of 52 lung SCC patients using survival analysis. The meta-analysis identified 20 consensus candidates, among which four (SENP2, DCUN1D1, DVL3, and UBXN7) are involved in a small protein-protein interaction network. Knocking down any of the four proteins led to cell growth inhibition of the 3q26-29-amplified SCC. Moreover, knocking down of SENP2 resulted in the most significant cell growth inhibition and downregulation of DCUN1D1 and DVL3. Importantly, a gene expression signature composed of SENP2, DCUN1D1, and DVL3 stratified patients into subgroups with different response to adjuvant chemotherapy. Together, our findings show that SENP2, DCUN1D1, and DVL3 are candidate driver genes in the 3q26-29 amplicon of SCC, providing novel insights into the molecular mechanisms of disease progression and may have significant implication in the management of SCC of the lung.


Sequential in cis mutagenesis in vivo reveals various functions for CTCF sites at the mouse HoxD cluster.
Mammalian Hox gene clusters contain a range of CTCF binding sites. In addition to their importance in organizing a TAD border, which isolates the most posterior genes from the rest of the cluster, the positions and orientations of these sites suggest that CTCF may be instrumental in the selection of various subsets of contiguous genes, which are targets of distinct remote enhancers located in the flanking regulatory landscapes. We examined this possibility by producing an allelic series of cumulative in cis mutations in these sites, up to the abrogation of CTCF binding in the five sites located on one side of the TAD border. In the most impactful alleles, the global chromatin architecture of the locus was modified, yet not drastically, illustrating that CTCF sites located on one side of a strong TAD border are sufficient to organize at least part of this insulation. Spatial colinearity in the expression of these genes along the major body axis was nevertheless maintained, despite abnormal expression boundaries. In contrast, strong effects were scored in the selection of target genes responding to particular enhancers, leading to the misregulation of Hoxd genes in specific structures. Altogether, while most enhancer-promoter interactions can occur in the absence of this series of CTCF sites, the binding of CTCF in the Hox cluster is required to properly transform a rather unprecise process into a highly discriminative mechanism of interactions, which is translated into various patterns of transcription accompanied by the distinctive chromatin topology found at this locus. Our allelic series also allowed us to reveal the distinct functional contributions for CTCF sites within this Hox cluster, some acting as insulator elements, others being necessary to anchor or stabilize enhancer-promoter interactions, and some doing both, whereas they all together contribute to the formation of a TAD border. This variety of tasks may explain the amazing evolutionary conservation in the distribution of these sites among paralogous Hox clusters or between various vertebrates.


Facial visualizations of women's voices suggest a cross-modality preference for femininity.
Women with higher-pitched voices and more feminine facial features are commonly judged as being more attractive than are women with lower-pitched voices and less feminine faces, possibly because both features are affected by (age-related) variations in endocrine status. These results are primarily derived from investigations of perceptions of variations in single-modality stimuli (i.e., faces or voices) in samples of young adult women. In the present study we sought to test whether male and female perceptions of women's voices affect visual representations of facial femininity. Eighty men and women judged voice recordings of 10 young girls (11-15 years), 10 adult women (19-28 years) and 10 peri-/post-menopausal women (50-64 years) on age, attractiveness, and femininity. Another 80 men and women were asked to indicate the face they think each voice corresponded to using a video that gradually changed from a masculine looking male face into a feminine looking female face. Both male and female participants perceived voices of young girls and adult women to be significantly younger, more attractive and feminine than those of peri-/post-menopausal women. Hearing young girls' and adult women's voices resulted in both men and women selecting faces that differed markedly in apparent femininity from those associated with peri-/post-menopausal women's voices. Voices of young girls had the strongest effect on visualizations of facial femininity. Our results suggest a cross-modal preference for women's vocal and facial femininity, which depends on female age and is independent of the perceiver's sex.


Voluntary activation failure contributes more to plantar flexor weakness than antagonist coactivation and muscle atrophy in chronic stroke survivors.
The contributions of nervous system muscle activation and muscle atrophy to poststroke weakness have not been evaluated together in the same subject. Maximal voluntary contraction (MVC) torque, voluntary activation (twitch interpolation), and electromyographic (EMG) amplitude were determined bilaterally in the plantar flexors of seven chronic stroke survivors (40-63 yr, 24-51 mo poststroke). Volumes of the plantar flexor muscles were determined bilaterally with magnetic resonance imaging (MRI). The mean (±SD) contralesional (paretic) MVC torque was less than one-half of the ipsilesional leg: 56.7 ± 57.4 vs. 147 ± 35.7 Nm (P = 0.006). Contralesional voluntary activation was only 48 ± 36.9%, but was near complete in the ipsilesional leg, 97 ± 1.9% (P = 0.01). The contralesional MVC EMG amplitude (normalized to the maximum M-wave peak-to-peak amplitude) of the gastrocnemii and soleus were 36.0 ± 28.5 and 36.0 ± 31.0% of the ipsilesional leg. Tibialis anterior (TA) EMG coactivation was not different between the contralesional (23.2 ± 24.0% of TA MVC EMG) and ipsilesional side (12.3 ± 5.7%) (P = 0.24). However, TA EMG coactivation was excessive (71%) in one subject and accounted for ~8% of her weakness based on the estimated antagonist torque. Relative (%ipsilesional leg) plantar flexor and gastrocnemii volumes were 88 ± 6% (P = 0.004) and 76 ± 15% (P = 0.01), respectively. Interlimb volume differences of the soleus, deep plantar flexors, and peronei were not significant. Preferred walking speed (0.83 ± 0.33 m/s) was related to the contralesional MVC torque (r(2) = 0.57, P = 0.05, N = 7), but the two subjects with the greatest weakness walked faster than three others. Our findings suggest that plantar flexor weakness in mobile chronic stroke survivors reflects mostly voluntary activation failure, with smaller contributions from antagonist activity and atrophy.


Systemic Retinaldehyde Treatment Corrects Retinal Oxidative Stress, Rod Dysfunction, and Impaired Visual Performance in Diabetic Mice.
Diabetes appears to induce a visual cycle defect because rod dysfunction is correctable with systemic treatment of the visual cycle chromophore 11-cis-retinaldehyde. However, later studies have found no evidence for visual cycle impairment. Here, we further examined whether photoreceptor dysfunction is corrected with 11-cis-retinaldehyde. Because antioxidants correct photoreceptor dysfunction in diabetes, the hypothesis that exogenous visual chromophores have antioxidant activity in the retina of diabetic mice in vivo was tested. Rod function in 2-month-old diabetic mice was evaluated using transretinal electrophysiology in excised retinas and apparent diffusion coefficient (ADC) MRI to measure light-evoked expansion of subretinal space (SRS) in vivo. Optokinetic tracking was used to evaluate cone-based visual performance. Retinal production of superoxide free radicals, generated mostly in rod cells, was biochemically measured with lucigenin. Diabetic mice were systemically treated with a single injection of either 11-cis-retinaldehyde, 9-cis-retinaldehyde (a chromophore surrogate), or all-trans-retinaldehyde (the photoisomerization product of 11-cis-retinaldehyde). Consistent with previous reports, diabetes significantly reduced (1) dark-adapted rod photo responses (transretinal recording) by ∼18%, (2) rod-dominated light-stimulated SRS expansion (ADC MRI) by ∼21%, and (3) cone-dominated contrast sensitivity (using optokinetic tracking [OKT]) by ∼30%. Both 11-cis-retinaldehyde and 9-cis-retinaldehyde largely corrected these metrics of photoreceptor dysfunction. Higher-than-normal retinal superoxide production in diabetes by ∼55% was also significantly corrected following treatment with 11-cis-retinaldehyde, 9-cis-retinaldehyde, or all-trans-retinaldehyde. Collectively, data suggest that retinaldehydes improve photoreceptor dysfunction in diabetic mice, independent of the visual cycle, via an antioxidant mechanism.


SU-E-I-26: Estimation of Micro-Angiographic Fluoroscope (MAF) Gain Settings for Digital Subtraction Angiography (DSA) Based on the Fluoroscopic Exposure.
The MAF is a new high-resolution detector which is being clinically evaluated in neuro-vascular procedures. The detector contains a large-dynamic-range, high-sensitivity light image intensifier with variable gain. Since the MAF is a research prototype only partially integrated with the clinical system, x-ray technique parameters must be set manually. To improve workflow we developed an automatic method to estimate and set the proper LII voltage (MAF gain) for DSA acquisition based on the fluoroscopic parameters. The detector entrance exposure (XD) can be written as the x-ray tube output exposure (Xo) times an object attenuation factor and an inverse-square correction. If the object attenuation, scatter and distances are unchanged and the effect of x-ray kVp changes are neglected, then the DSA XD can be expressed as the ratio of Xo(DSA)/Xo(Fluoroscopy) multiplied with XD(fluoroscopy). We measured Xo for fluoroscopy and DSA for mAs and kVp ranges appropriate to neuro- vascular interventions and fit the data with a 2D function. To estimate the XD(Fluoroscopy) we derived a curve of XD versus LII-voltage for a mid- dynamic-range average pixel gray-level. Since the MAF system during clinical fluoroscopy automatically adjusts the LII voltage until the desired gray-level value is achieved, by reading that voltage we can estimate the XD(Fluoroscopy). Using the 2D-fit function, Xo(DSA) is automatically calculated for the kVp and mA values set and XD(DSA) can be estimated using the relation above. Using the inverse LII calibration curve, the proper LII-voltage can be determined for the desired average gray-level. The algorithm was implemented and evaluated in thirty-two in-vivo DSA runs on rabbits. The proper LII voltage was selected in all cases with no failures. Using the fluoroscopic LII gain setting to determine the appropriate DSA setting can greatly improve the workflow in clinical evaluations of the MAF. NIH Grants R01-EB008425, R01-EB002873 and an equipment grant from Toshiba Medical Systems Corp.


Update on systemic vasculitides.
Management of systemic vasculitis has been revolutionised over the last decade with the introduction of targeted biological agents. With an increase in both the prevalence and the recognition of vasculitis as well as the high cost of these agents, it is important to ensure their most optimal utilisation. The goals of vasculitis therapy include the induction and maintenance of remissions, preventing relapses, reducing the toxicity of therapy with the aim of reducing morbidity and mortality as well as improving the quality of life of those afflicted. This review focuses on the recent advances in the diagnosis, surveillance and treatment of these conditions.


Significance of wine and resveratrol in cardiovascular disease: French paradox revisited.
Many recent studies have reported promising health benefits from red wine consumption. The present article reviews some of the key studies, and the known mechanisms for these beneficial effects. Evidence from different experimental studies, including from the authors' laboratories, have suggested that these beneficial effects are due to polyphenols found in red wine, especially resveratrol in grape skins. These benefits include a reduction in cardiovascular morbidity and mortality, lung cancer and prostate cancer by approximately 30% to 50%, 57% and 50%, respectively. Polyphenols possess antioxidant, superoxide-scavenging, ischemic-preconditioning and angiogenic properties. Some of these properties of polyphenols may explain their protective effects on the cardiovascular system, as well as other body organs. In fact, results from several epidemiological, case-control and prospective studies have prompted the United States Department of Health and Human Services to recommend moderate alcohol consumption in its national health promotion and disease prevention initiative, Healthy People 2010. Further studies are warranted to describe the precise molecular mechanisms for these potential beneficial effects of red wine on the general health of the population, particularly on cardiovascular morbidity and mortality.


Root ethylene mediates rhizosphere microbial community reconstruction when chemically detecting cyanide produced by neighbouring plants.
Stress-induced hormones are essential for plants to modulate their microbiota and dynamically adjust to the environment. Despite the emphasis of the role of the phytohormone ethylene in the plant physiological response to heterospecific neighbour detection, less is known about how this activated signal mediates focal plant rhizosphere microbiota to enhance plant fitness. Here, using 3 years of peanut (Arachis hypogaea L.), a legume, and cyanide-containing cassava (Manihot esculenta Crantz) intercropping and peanut monocropping field, pot and hydroponic experiments in addition to exogenous ethylene application and soil incubation experiments, we found that ethylene, a cyanide-derived signal, is associated with the chemical identification of neighbouring cassava and the microbial re-assemblage in the peanut rhizosphere. Ethylene production in peanut roots can be triggered by cyanide production of neighbouring cassava plants. This gaseous signal alters the microbial composition and re-assembles the microbial co-occurrence network of peanut by shifting the abundance of an actinobacterial species, Catenulispora sp., which becomes a keystone in the intercropped peanut rhizosphere. The re-assembled rhizosphere microbiota provide more available nutrients to peanut roots and support seed production. Our findings suggest that root ethylene acts as a signal with a dual role. It plays a role in perceiving biochemical cues from interspecific neighbours, and also has a regulatory function in mediating the rhizosphere microbial assembly, thereby enhancing focal plant fitness by improving seed production. This discovery provides a promising direction to develop novel intercropping strategies for targeted manipulations of the rhizosphere microbiome through phytohormone signals. Video abstract.


Role of diffusion MRI and apparent diffusion coefficient measurement in the diagnosis, staging and pathological classification of bladder tumors.
Our aim was to investigate the role of simultaneous apparent diffusion coefficient (ADC) measurement and diffusion-weighted MRI (DWMRI) in the diagnosis and differentiation of bladder masses. We examined 45 patients presenting with hematuria by dynamic and diffusion MRI (at b100, b600, b1000 gradients) and cystoscopy. Thirty healthy volunteers without hematuria were also included in the study. Results from imaging techniques were compared to cystoscopic and histopathologic findings. The ADC values of benign and malignant bladder masses, and bladder tissue in healthy subjects were measured and compared with each other. The mean ADC values of benign (n = 10) and malignant lesions (n = 35) were significantly lower than the mean ADC values of adjacent bladder walls and bladder walls of healthy subjects at all 3 gradients (p < 0.05). However, no significant difference was detected between benign and malignant lesions (p > 0.05). Mean ADC values of invasive tumors were significantly lower than superficial malignancies at b600 and b1000 gradients. The mean ADC values of transitional cell tumors were significantly higher than squamous cell tumors at b100 and b600 gradients. Simultaneous measurement ofADC and DWMRI provided useful information in the diagnosis, staging and histopathological classification of bladder tumors.


Global biodiversity: indicators of recent declines.
In 2002, world leaders committed, through the Convention on Biological Diversity, to achieve a significant reduction in the rate of biodiversity loss by 2010. We compiled 31 indicators to report on progress toward this target. Most indicators of the state of biodiversity (covering species' population trends, extinction risk, habitat extent and condition, and community composition) showed declines, with no significant recent reductions in rate, whereas indicators of pressures on biodiversity (including resource consumption, invasive alien species, nitrogen pollution, overexploitation, and climate change impacts) showed increases. Despite some local successes and increasing responses (including extent and biodiversity coverage of protected areas, sustainable forest management, policy responses to invasive alien species, and biodiversity-related aid), the rate of biodiversity loss does not appear to be slowing.


Survival Benefit of Tolvaptan for Refractory Ascites in Patients with Advanced Cirrhosis.
The study aimed to evaluate the effects of tolvaptan treatment on survival of patients with decompensated liver cirrhosis with refractory ascites. This multicenter, retrospective, observational study included patients with cirrhosis who were treated with tolvaptan for hepatic ascites refractory to conventional diuretics. Patients who could and could not decrease accompanying diuretics within 1 month after tolvaptan administration were defined as the "Decreased" and "Not-decreased" groups, respectively. Median body weight change 1 week after tolvaptan treatment was -1.95 kg, with the 50% of patients experiencing a 2 kg/week reduction. Spot urinary sodium was found to be a better predictor of tolvaptan response than liver function and liver fibrosis markers. Median survival was significantly longer (not reached versus 116 days, p = 0.005) and serum creatinine concentrations 12 weeks after tolvaptan administration significantly lower (0.99 vs. 1.55 mg/dL, p < 0.05) in the Decreased than in the Not-decreased group. Multivariate analysis showed that the presence of viable hepatocellular carcinoma (hazards ratio [HR] 2.14, p = 0.02) and a decrease in diuretics were independently prognostic of survival (HR 0.36, p < 0.01). The maintenance of renal function is essential in enhancing survival of patients with cirrhosis. Doses of diuretics should be adjusted appropriately during tolvaptan treatment.


The role of free radicals and plasmatic antioxidant in Ménière's syndrome.
The objective of this study was verification, through suitable hematochemical tests, of the supposition that central-systemic microtoxicosis plays a role either in the etiopathogenesis of Ménière's syndrome or in labyrinthine pathological processes or hypoacusis. We did not, therefore, exclude other well-known hypotheses in the causality of these pathologies. Nonetheless, one finds, particularly in the Ménière's cases, a constant homogeneous distribution of the metabolic products of this microtoxicosis, such as a high concentration of free radicals and low natural defenses (e.g., antioxidant plasmatic capacity). Therefore, there exists a kind of dangerous central and systemic presence of reactive molecules, aimed toward the polyunsaturated fatty acids and homeostatic complex enzymes, that is not compensated for by the natural antioxidant defense. The presence of this lack of balance, verified by suitable tests, has shown the rationality of use of a product made from reduced glutathione, thioctic acid, cysteine, and other antioxidants as a multipurpose antidote to this element of etiopathogenesis. Patients were divided into three groups (control, conventional therapy, and antioxidant treatment), and those in the antioxidant treatment group, especially those with Ménière's syndrome, demonstrated a net and more significant improvement. Also, parallel clinical and instrument evaluations of this new therapeutic solution, the efficacy of which has already been positively demonstrated, are expected to provide further evidence to support the primary hypothesis.


Detecting cancer biomarkers in blood: challenges for new molecular diagnostic and point-of-care tests using cell-free nucleic acids.
As we move into the era of individualized cancer treatment, the need for more sophisticated cancer diagnostics has emerged. Cell-free (cf) nucleic acids (cf-DNA, cf-RNA) and other cellular nanoparticulates are now considered important and selective biomarkers. There is great hope that blood-borne cf-nucleic acids can be used for 'liquid biopsies', replacing more invasive tissue biopsies to analyze cancer mutations and monitor therapy. Conventional techniques for cf-nucleic acid biomarker isolation from blood are generally time-consuming, complicated and expensive. They require relatively large blood samples, which must be processed to serum or plasma before isolation of biomarkers can proceed. Such cumbersome sample preparation also limits the widespread use of powerful, downstream genomic analyses, including PCR and DNA sequencing. These limitations also preclude rapid, point-of-care diagnostic applications. Thus, new technologies that allow rapid isolation of biomarkers directly from blood will permit seamless sample-to-answer solutions that enable next-generation point-of-care molecular diagnostics.


Reasoning as we read: establishing the probability of causal conditionals.
Indicative conditionals of the form if p then q (e.g., if student tuition fees rise, then applications for university places will fall) invite consideration of a hypothetical event (e.g., tuition fees rising) and of one of its possible consequences (e.g., applications falling). Since a rise in tuition fees is an uncertain event with equally uncertain consequences, a reader may believe the statement to a greater or lesser extent. As a conditional is read, the earliest point at which this probabilistic evaluation can take place is as the consequent clause is wrapped up (e.g., as the critical word fall is read in the example above). Wrap-up processing occurs at the end of the clause, as it is evaluated and integrated into the evolving discourse representation. Five sources of probability may plausibly influence the evaluation of a conditional as it is wrapped up; these are P(p), P(q), P(pq), P(q|p), and P(not-p or q). A total of 128 conditionals were constructed, with these probabilities calculated for each item in a pretest. The conditionals were then embedded in vignettes and read by 36 participants on a word-by-word basis. Using linear mixed-effects modeling, we found that wrap-up reading times were predicted by pretest ratings of P(p) and P(q|p). There was no influence of P(q), P(pq), or P(not-p or q) on wrap-up reading times. Our findings are consistent with the suppositional theory of conditionals proposed by Evans and Over (2004) but do not support the mental-models theory advanced by Johnson-Laird and Byrne (2002).


Overexpression of the csA cell adhesion molecule under its own cAMP-regulated promoter impairs morphogenesis in Dictyostelium.
The contact site A (csA) glycoprotein is a strictly developmentally regulated plasma membrane component responsible for the EDTA-stable (Ca(2+)-independent) form of intercellular adhesion in Dictyostelium discoideum. Using inverse polymerase chain reaction and a terminator vector we have isolated a 1.6 kb genomic fragment carrying a 1.1 kb upstream region of the csA gene. This fragment had promoter activity in D. discoideum cells, giving rise to a 3'-truncated csA RNA that was regulated like the mRNA of the endogenous gene. Cyclic AMP pulses strongly enhanced transcription from the cloned csA promoter. These findings provide evidence that the cloned region of the csA gene comprises the complete promoter. It contains a G/C-rich octamer motif similar to other cAMP-regulated D. discoideum promoters. When the csA protein was strongly overexpressed under the developmental control of the csA promoter, morphogenesis was substantially altered. Aggregation was delayed, and secondary centres were formed along aggregation streams that led to fragmentation of the aggregates and multiple slug formation. At high cell density a substantial portion of aggregated cells was left behind on the substratum when slugs and fruiting bodies were built. The transformation vector was also employed to rescue a csA-negative mutant, HG1287, from its cell adhesion defect.


Why does economic analysis in health care not get implemented more? Towards a greater understanding of the rules of the game and the costs of decision making.
Despite recent initiatives across a number of countries to institutionalise the use of economic evaluation in the assessment of new technologies, the evidence suggests that, to date, the impact of economic analysis on health sector decision making in practice has been low. This paper proposes that incentive compatibility problems associated with much of the decision making that economics is meant to inform is a major factor in this. The argument involves recognition that economic priority setting tends to create winners and losers and, as a consequence, any change in the status quo will tend to impose direct costs on those decision makers required to implement them. At the same time, such individuals often lack any stake over the benefits of such decisions. In this type of institutional setting, the prospects of achieving the necessary cooperation for implementation are likely to be low, particularly if it is viewed as being part of a 'one-off' initiative. Conventional health economic analysis and agency theory are limited in the extent to which they can address this problem. Such forms of analyses tend to lack an appreciation of broader institutional characteristics, including the prevailing values and norms within a decision making context that are potentially important in promoting cooperation and thus the prospects of implementation.


Biomimetic super-lyophobic and super-lyophilic materials applied for oil/water separation: a new strategy beyond nature.
Oil spills and industrial organic pollutants have induced severe water pollution and threatened every species in the ecological system. To deal with oily water, special wettability stimulated materials have been developed over the past decade to separate oil-and-water mixtures. Basically, synergy between the surface chemical composition and surface topography are commonly known as the key factors to realize the opposite wettability to oils and water and dominate the selective wetting or absorption of oils/water. In this review, we mainly focus on the development of materials with either super-lyophobicity or super-lyophilicity properties in oil/water separation applications where they can be classified into four kinds as follows (in terms of the surface wettability of water and oils): (i) superhydrophobic and superoleophilic materials, (ii) superhydrophilic and under water superoleophobic materials, (iii) superhydrophilic and superoleophobic materials, and (iv) smart oil/water separation materials with switchable wettability. These materials have already been applied to the separation of oil-and-water mixtures: from simple oil/water layered mixtures to oil/water emulsions (including oil-in-water emulsions and water-in-oil emulsions), and from non-intelligent materials to intelligent materials. Moreover, they also exhibit high absorption capacity or separation efficiency and selectivity, simple and fast separation/absorption ability, excellent recyclability, economical efficiency and outstanding durability under harsh conditions. Then, related theories are proposed to understand the physical mechanisms that occur during the oil/water separation process. Finally, some challenges and promising breakthroughs in this field are also discussed. It is expected that special wettability stimulated oil/water separation materials can achieve industrial scale production and be put into use for oil spills and industrial oily wastewater treatment in the near future.


Introductory lecture: recent research progress on aggregation-induced emission.
Since the discovery of the aggregation-induced emission (AIE) phenomenon in 2001, research on AIE molecules has drawn much attention, and this area has been expanding tremendously. This brief review will focus on recent advances in the science and application of AIE molecules, including new mechanistic understanding, new AIE molecules for sensing and imaging, stimuli-responsive AIE molecules and applications of AIE molecules for OLEDs. Moreover, this review will give a perspective on the possible opportunities and challenges that exist in the future for this area.


CFTR modulator theratyping: Current status, gaps and future directions.
New drugs that improve the function of the cystic fibrosis transmembrane conductance regulator (CFTR) protein with discreet disease-causing variants have been successfully developed for cystic fibrosis (CF) patients. Preclinical model systems have played a critical role in this process, and have the potential to inform researchers and CF healthcare providers regarding the nature of defects in rare CFTR variants, and to potentially support use of modulator therapies in new populations. The Cystic Fibrosis Foundation (CFF) assembled a workshop of international experts to discuss the use of preclinical model systems to examine the nature of CF-causing variants in CFTR and the role of in vitro CFTR modulator testing to inform in vivo modulator use. The theme of the workshop was centered on CFTR theratyping, a term that encompasses the use of CFTR modulators to define defects in CFTR in vitro, with application to both common and rare CFTR variants. Several preclinical model systems were identified in various stages of maturity, ranging from the expression of CFTR variant cDNA in stable cell lines to examination of cells derived from CF patients, including the gastrointestinal tract, the respiratory tree, and the blood. Common themes included the ongoing need for standardization, validation, and defining the predictive capacity of data derived from model systems to estimate clinical outcomes from modulator-treated CF patients. CFTR modulator theratyping is a novel and rapidly evolving field that has the potential to identify rare CFTR variants that are responsive to approved drugs or drugs in development.


Dissolution of ibuprofen from spray dried and spray chilled particles.
The formulation of hydrophobic drugs for oral drug delivery is challenging due to poor solubility, poor dissolution and poor wetting of these drugs. Consequently, the aim of this study was to improve the dissolution of a model poorly water soluble drug, ibuprofen. Microparticles containing ibuprofen were produced by spray drying and spray chilling technology in the absence/presence of a hydrophilic surfactant. Poloxamer 127, tri-block copolymer, was chosen as the hydrophilic surfactant to improve drug particle wettability and hence the dissolution rate. The prepared formulations were evaluated for in vitro dissolution and intrinsic solubility. In addition, the produced drug particles were characterised by scanning electron microscopy (SEM), differential scanning calorimeter (DSC) and Fourier transform infrared spectroscopy (FT-IR). SEM revealed changes in the surface morphology of processed ibuprofen, suggesting the effective formation of the drug particles. DSC data showed shifting of the melting peak of the drug towards lower melting temperature in the prepared particles, indicating the possibility of drug /polymer interaction. The results of the dissolution studies of spray dried ibuprofen and spray dried ibuprofen/Poloxamer 127 particles showed significantly (P<0.05) increased percentage drug release compared to control (ibuprofen raw material). For spray chilling, the prepared particles did not improve the dissolution of the drug, the dissolution was even less than that of the control. DSC and FT-IR results demonstrated that spray drying reduced drug crystallinity, but for spray chilled particles there was evidence of polymorphic changes in the drug with and without the surfactant. Consequently, it is believed that spray drying of ibuprofen is a useful tool to improve wettability, solubility and hence the dissolution behaviour of poorly water soluble drugs, in contrast to spray chilling technique.


Intraoperative blood loss during different stages of scoliosis surgery: A prospective study.
There are a number of reasons for intraoperative blood loss during scoliosis surgery based on the type of approach, type of disease, osteopenia, and patient blood profile. However, no studies have investigated bleeding patterns according to the stage of the operation. The objective of this prospective study was to identify intraoperative bleeding patterns in different stages of scoliosis surgery. We prospectively analyzed the estimated blood loss (EBL) and operation time over four stages of scoliosis surgery in 44 patients. The patients were divided into three groups: adolescent idiopathic (group 1), spastic neuromuscular (group 2) and paralytic neuromuscular (group 3). The per-level EBL and operation times of the groups were compared on a stage-by-stage basis. The bone marrow density (BMD) of each patient was also obtained, and the relationship between per-level EBL and BMD was compared using regression analysis. Per-level operation time was similar across all groups during surgical stage (p > 0.05). Per-level EBL was also similar during the dissection and bone-grafting states (p > 0.05). However, during the screw insertion stage, the per-level EBL was significantly higher in groups 2 and 3 compared to group 1 (p < 0.05). In the correction stage, per-level EBL was highest in group 3 (followed in order by groups 2 and 1) (p < 0.05). Preoperative BMD indicated that group 3 had the lowest bone quality, followed by groups 2 and 1 (in order), but the preoperative blood indices were similar in all groups. The differences in bleeding patterns in the screw insertion and correction stages were attributed to the poor bone quality of groups 2 and 3. Group 3 had the lowest bone quality, which caused loosening of the bone-screw interface during the correction stage and led to more bleeding. Patients with a T-score less than -2.5 showed a risk for high per-level EBL that was nine times higher than those with scores greater than -2.5 (p = 0.003). We investigated the blood loss patterns during different stages of scoliosis surgery. Patients with poor BMD showed a risk of blood loss nine times higher than those with good BMD.


Low CO2 Sensitivity of Microzooplankton Communities in the Gullmar Fjord, Skagerrak: Evidence from a Long-Term Mesocosm Study.
Ocean acidification is considered as a crucial stressor for marine communities. In this study, we tested the effects of the IPCC RPC6.0 end-of-century acidification scenario on a natural plankton community in the Gullmar Fjord, Sweden, during a long-term mesocosm experiment from a spring bloom to a mid-summer situation. The focus of this study was on microzooplankton and its interactions with phytoplankton and mesozooplankton. The microzooplankton community was dominated by ciliates, especially small Strombidium sp., with the exception of the last days when heterotrophic dinoflagellates increased in abundance. We did not observe any effects of high CO2 on the community composition and diversity of microzooplankton. While ciliate abundance, biomass and growth rate were not affected by elevated CO2, we observed a positive effect of elevated CO2 on dinoflagellate abundances. Additionally, growth rates of dinoflagellates were significantly higher in the high CO2 treatments. Given the higher Chlorophyll a content measured under high CO2, our results point at mainly indirect effects of CO2 on microzooplankton caused by changes in phytoplankton standing stocks, in this case most likely an increase in small-sized phytoplankton of <8 μm. Overall, the results from the present study covering the most important part of the growing season indicate that coastal microzooplankton communities are rather robust towards realistic acidification scenarios.


Relationship Between Treatment Initiation and Healthcare Costs in Alzheimer's Disease.
The relationship between Alzheimer's disease (AD) treatment patterns and healthcare costs is unknown. Administrative claims data from the MarketScan Commercial and Medicare databases covering 2010 through 2016 were used to identify the comorbidities, treatment patterns, and healthcare costs in the three years prior to and one year post medical diagnosis of AD in 21,448 patients with no treatment and 57,970 patients with treatment. Pre-index mean annual costs ranged from $14,228 to $26,876, and post-index mean annual costs ranged from $21,052 to $45,685 depending on age and treatment timing. After adjusting for baseline characteristics, patients 50-100 years old who initiated treatment with an FDA approved drug prior to or concurrent with diagnosis had healthcare costs 9%-19% lower in the year following diagnosis than those who did not receive treatment. Early or concurrent treatment is associated with lower overall healthcare costs in the year following AD diagnosis.


Characterization of Environmental Dust in the Dammam Area and Mud After-Effects on Bisphenol-A Polycarbonate Sheets.
Owing to recent climate changes, dust storms are increasingly common, particularly in the Middle East region. Dust accumulation and subsequent mud formation on solid surfaces in humid environments typically have adverse effects on surface properties such as optical transmittance, surface texture, and microhardness. This is usually because the mud, which contains alkaline and ionic species, adheres strongly to the surface, often through chemical bonds, and is therefore difficult to remove. In this study, environmental dust and the after-effects of mud formed on a polycarbonate sheet, which is commonly used as a protective glass in photovoltaic cells. Ionic compounds (OH(-)) are shown to significantly affect the optical, mechanical, and textural characteristics of the polycarbonate surface, and to increase the adhesion work required to remove the dry mud from the polycarbonate surface upon drying. Such ability to modify characteristics of the polycarbonate surface could address the dust/mud-related limitations of superhydrophobic surfaces.


Inflammasome signaling and regulation of interleukin-1 family cytokines.
Specific IL-1 family cytokines are expressed by cells as cytosolic pro-forms that require cleavage for their activity and cellular release. IL-1β, IL-18, and IL-37 maturation and secretion is governed by inflammatory caspases within signaling platforms called inflammasomes. By inducing pyroptosis, inflammasomes can also drive the release of the alarmin IL-1α. Recent advances have transformed our mechanistic understanding of inflammasome signaling, cell death decisions, and cytokine activation and secretion. Here, we provide an updated view of inflammasome signaling; mechanisms underpinning IL-1α, IL-1β, IL-18, and IL-37 maturation and release; and the functions of these cytokines in protective and pathological inflammation.


Relationship between Cognitive Dysfunction and Age-Related Variability in Oxidative Markers in Isolated Mitochondria of Alzheimer's Disease Transgenic Mouse Brains.
Many neurodegenerative disorders, including Alzheimer's disease (AD), are strongly associated with the accumulation of oxidative damage. Transgenic animal models are commonly used to elucidate the pathogenic mechanism of AD. Beta amyloid (Aβ) and tau hyperphosphorylation are very famous hallmarks of AD and well-studied, but the relationship between mitochondrial dysfunction and the onset and progression of AD requires further elucidation. In this study we used transgenic mice (the strain name is 5xFAD) at three different ages (3, 6, and 20 months old) as an AD model. Cognitive impairment in AD mice occurred in an age-dependent manner. Aβ1-40 expression significantly increased in an age-dependent manner in all brain regions with or without AD, and Aβ1-42 expression in the hippocampus increased at a young age. In a Western blot analysis using isolated mitochondria from three brain regions (cerebral cortex, cerebellum, and hippocampus), NMNAT-3 expression in the hippocampi of aged AD mice was significantly lower than that of young AD mice. SOD-2 expression in the hippocampi of AD mice was lower than for the age-matched controls. However, 3-NT expression in the hippocampi of AD mice was higher than for the age-matched controls. NQO-1 expression in the cerebral cortex of AD mice was higher than for the age-matched controls at every age that we examined. However, hippocampal NQO-1 expression in 6-month-old AD mice was significantly lower than in 3-month-old AD mice. These results indicate that oxidative stress in the hippocampi of AD mice is high compared to other brain regions and may induce mitochondrial dysfunction via oxidative damage. Protection of mitochondria from oxidative damage may be important to maintain cognitive function.


Use of immunotoxins in combination to inhibit clonogenic growth of human breast carcinoma cells.
Substantial heterogeneity has been observed in the expression of individual antigens within tumor cell populations. Immunotoxins which bind to different cell surface antigens might exert additive or synergistic cytotoxicity when used in combination to eliminate all clonogenic cells within a tumor. Immunotoxins have been prepared by conjugating recombinantly derived toxin A chain to different monoclonal reagents which recognize cell surface determinants of Mr 42,000 (317G5), 55,000 (260F9), and 200,000 (741F8). Each immunotoxin was evaluated for binding, internalization, and cytotoxicity with four breast cancer cell lines. Each of the three immunotoxins bound to the SKBr3 cell line and exerted antitumor activity in a limiting dilution clonogenic assay. Simultaneous treatment with two immunotoxins produced additive antitumor activity with each of the possible combinations. Additive binding could be demonstrated by immunofluorescent techniques, however, with only one of three combinations. With two of the three combinations, subpopulations of tumor cells could be identified which lacked one or the other antigenic determinant but not both. Consequently, log-additive antitumor activity was produced by immunotoxins in combination, and heterogeneity of antigenic targets may have contributed to the combined cytotoxicity in some but not all cases.


Analysing the floral elements of the lost tree of Easter Island: a morphometric comparison between the remaining ex-situ lines of the endemic extinct species Sophora toromiro.
Sophora toromiro (Phil) Skottsb. is a species that has been extinct in its natural habitat Easter Island (Rapa Nui) for over 50 years. However, seed collections carried out before its extinction have allowed its persistence ex-situ in different botanical gardens and private collections around the world. The progenies of these diverse collections have been classified in different lines, most of them exhibiting high similarity as corroborated by molecular markers. In spite of this resemblance observed between the different lines, one of them (Titze) has dissimilar floral elements, thus generating doubts regarding its species classification. The floral elements (wing, standard and keel) belonging to three different S. toromiro lines and two related species were analyzed using geometric morphometrics. This method was applied in order to quantify the floral shape variation of the standard, wing, and keel between the different lines and control species. Geometric morphometrics analyses were able to distinguish the floral elements at both intra (lines) and inter-specific levels. The present results are on line with the cumulative evidence that supports the Titze line as not being a proper member of the S. toromiro species, but probably a hybridization product or even another species of the Edwardsia section. The reintroduction programs of S. toromiro should consider this information when assessing the authenticity and origin of the lines that will be used to repopulate the island.


Comparing error minimized extreme learning machines and support vector sequential feed-forward neural networks.
Recently, error minimized extreme learning machines (EM-ELMs) have been proposed as a simple and efficient approach to build single-hidden-layer feed-forward networks (SLFNs) sequentially. They add random hidden nodes one by one (or group by group) and update the output weights incrementally to minimize the sum-of-squares error in the training set. Other very similar methods that also construct SLFNs sequentially had been reported earlier with the main difference that their hidden-layer weights are a subset of the data instead of being random. These approaches are referred to as support vector sequential feed-forward neural networks (SV-SFNNs), and they are a particular case of the sequential approximation with optimal coefficients and interacting frequencies (SAOCIF) method. In this paper, it is firstly shown that EM-ELMs can also be cast as a particular case of SAOCIF. In particular, EM-ELMs can easily be extended to test some number of random candidates at each step and select the best of them, as SAOCIF does. Moreover, it is demonstrated that the cost of the computation of the optimal output-layer weights in the originally proposed EM-ELMs can be improved if it is replaced by the one included in SAOCIF. Secondly, we present the results of an experimental study on 10 benchmark classification and 10 benchmark regression data sets, comparing EM-ELMs and SV-SFNNs, that was carried out under the same conditions for the two models. Although both models have the same (efficient) computational cost, a statistically significant improvement in generalization performance of SV-SFNNs vs. EM-ELMs was found in 12 out of the 20 benchmark problems.


Serum 25-hydroxyvitamin D concentrations in relation to cardiometabolic risk factors and metabolic syndrome in postmenopausal women.
Low concentrations of serum 25-hydroxyvitamin D [25(OH)D] may be associated with cardiometabolic disorders; however, little is known about their relation to intermediate metabolic and lipid markers. We investigated the relation of serum 25(OH)D concentrations to fasting insulin, glucose, dyslipidemia, adiposity, and prevalent metabolic syndrome. We conducted this cross-sectional analysis in 292 postmenopausal women aged 50-79 y in the Women's Health Initiative Calcium-Vitamin D (WHI-CaD) trial. Data were collected from 3 nested case-control studies that measured baseline serum 25(OH)D concentrations. Inverse probability weighting was used to approximate parameter estimates for the WHI-CaD population. In weighted linear regression models adjusted for age, race-ethnicity, month of blood draw, region, case-control status, smoking, alcohol, physical activity, and history of cardiometabolic risk factors, there was an inverse association of serum 25(OH)D with adiposity [body mass index (BMI): β = -1.12 ± 0.30, P = 0.0002; waist circumference: β = -3.57 ± 0.49, P < 0.0001; waist-hip ratio: β = -0.01 ± 0.002, P < 0.0001], triglycerides (β = -0.10 ± 0.02, P < 0.0001), and triglyceride:HDL-cholesterol ratio (β = -0.11 ± 0.03, P = 0.0003). The multivariable-adjusted odds ratio for metabolic syndrome for the highest (≥52 nmol/L) compared with the lowest (<35 nmol/L) tertile of serum 25(OH)D concentrations was 0.28 (95% CI: 0.14, 0.56). Significant associations remained after adjustment for BMI. We observed no significant associations with LDL cholesterol, HDL cholesterol, insulin, glucose, homeostatic model assessment of insulin resistance (HOMA-IR), or homeostatic model assessment of β cell function (HOMA-β). Higher serum 25(OH)D concentrations may be inversely associated with adiposity, triglycerides, triglyceride:HDL-cholesterol ratio, and metabolic syndrome but are not associated with LDL and HDL cholesterol, insulin, glucose, HOMA-IR, or HOMA-β in postmenopausal women. This trial was registered at clinicaltrials.gov as NCT00000611.


Genome-wide association study for circulating levels of PAI-1 provides novel insights into its regulation.
We conducted a genome-wide association study to identify novel associations between genetic variants and circulating plasminogen activator inhibitor-1 (PAI-1) concentration, and examined functional implications of variants and genes that were discovered. A discovery meta-analysis was performed in 19 599 subjects, followed by replication analysis of genome-wide significant (P < 5 × 10(-8)) single nucleotide polymorphisms (SNPs) in 10 796 independent samples. We further examined associations with type 2 diabetes and coronary artery disease, assessed the functional significance of the SNPs for gene expression in human tissues, and conducted RNA-silencing experiments for one novel association. We confirmed the association of the 4G/5G proxy SNP rs2227631 in the promoter region of SERPINE1 (7q22.1) and discovered genome-wide significant associations at 3 additional loci: chromosome 7q22.1 close to SERPINE1 (rs6976053, discovery P = 3.4 × 10(-10)); chromosome 11p15.2 within ARNTL (rs6486122, discovery P = 3.0 × 10(-8)); and chromosome 3p25.2 within PPARG (rs11128603, discovery P = 2.9 × 10(-8)). Replication was achieved for the 7q22.1 and 11p15.2 loci. There was nominal association with type 2 diabetes and coronary artery disease at ARNTL (P < .05). Functional studies identified MUC3 as a candidate gene for the second association signal on 7q22.1. In summary, SNPs in SERPINE1 and ARNTL and an SNP associated with the expression of MUC3 were robustly associated with circulating levels of PAI-1.


The alchemy of tendon repair: a primer for the (S)mad scientist.
During vertebrate development, mesenchymal progenitors capable of forming bone, cartilage, muscle, fat, or tendon arise from either neural crest or somitic mesoderm. Transcriptional programs that specify mesenchymal cell fates are initiated and modified by paracrine cues provided by TGF-beta superfamily members and mediated in part via the regulated assembly of Smad-containing multiprotein transcription factor complexes. In this issue of the JCI, Hoffmann and colleagues have identified that Smad8 activation drives tendon formation from C3H10T1/2 cells, a murine cell line that recapitulates many features of normal multipotent mesenchymal cells (see the related article beginning on page 940). Cells programmed to the tenocyte cell fate in vitro formed tenogenic grafts in vivo. These results add to the accumulating evidence that proliferating, multipotent mesenchymal progenitor cells can be programmed to yield multiple cell types--e.g., osteoblasts, myocytes, chondrocytes, and tenocytes--that may be useful in cell-based therapeutic approaches to musculoskeletal diseases.


Can father inclusive practice reduce paternal postnatal anxiety? A repeated measures cohort study using the Hospital Anxiety and Depression Scale.
Perinatal research on anxiety and depression has primarily focused on mothers. We have limited knowledge of fathers' anxiety during the perinatal period yet there is evidence that the parenting capacity of a person can be compromised by anxiety and depression. The purpose of this paper is to identify the impact of a father inclusive intervention on perinatal anxiety and depression. The prime focus of the intervention was to provide education and support to fathers of breastfeeding partners with the aim of increasing both initiation and duration of breastfeeding. A repeated measures cohort study was conducted during a RCT that was implemented across eight public maternity hospitals in Perth, Western Australia between May 2008 and June 2009. A baseline questionnaire which included the Hospital Anxiety and Depression Scale (HADS) was administered to all participants on the first night of their hospital based antenatal education program and was repeated at six weeks postnatal. SPSS version 17 was used for reporting descriptive results. The mean anxiety levels at baseline for the fathers in the intervention group (n=289) and control group (n=244) were 4.58 and 4.22 respectively. At 6 weeks postnatal (only matched pairs), intervention and control group were 3.93 and 3.79. More intervention group fathers self-rated less anxiety compared to the fathers in the control group from baseline to post test (p=0.048). Depression scores for intervention fathers at baseline (mean=1.09) and at six weeks (mean=1.09) were very similar to fathers in the control group at baseline (mean=1.11) and at six weeks (mean=1.07) with no significant changes. Both intervention and control group fathers experienced some anxiety prior to the birth of their baby, but this was rapidly reduced at six weeks. Paternal anxiety is common to new fathers and providing them with information and strategies for problem-solving can increase their knowledge and potentially lower the risk of postnatal anxiety. (Australian New Zealand Clinical Trials Registry ACTRN12609000667213).


High-performance all-solid-state flexible micro-supercapacitor arrays with layer-by-layer assembled MWNT/MnO(x) nanocomposite electrodes.
In this study, we report on the fabrication of high performance planar-type flexible micro-supercapacitor (MSC) arrays using Au electrodes coated with a functionalized multi-walled carbon nanotube (MWNT) film and a layer of MWNT-COOH/MnOx nanoparticle (NP) composite on top. The MWNT thin film was formed via layer-by-layer (LbL) assembly of MWNTs functionalized with amine groups and MWNTs with carboxylic acid groups in water. The hydrothermally synthesized composite of MWNT-COOH/MnOx NPs was coated on top of the MWNT film (LbL-MWNT). The addition of MWNT-COOH/MnOx NP composite as a top layer enhanced the performance of the MSCs dramatically, resulting in a volumetric capacitance of 50 F cm(-3) at a scan rate of 10 mV s(-1) and a coulombic efficiency of ∼100%. By contrast, a volumetric capacitance of 3.6 F cm(-3) was obtained when using only the LbL-MWNT film. After repetitive operation up to ∼10(4) times, the capacitance remained at ∼88.3% of the original value. With a deliberate circuit design consisting of serially connected MSC arrays, various light-emitting diodes operating at different bias voltages could be lit. The MSC circuit fabricated on a polyethylene terephthalate (PET) film showed stable electrochemical properties upon 1000 cycles of bending deformation.


15-Year prognostic utility of coronary artery calcium scoring for all-cause mortality in the elderly.
Prior studies have demonstrated a decline in the predictive ability of conventional risk factors (RF) with advancing age, emphasizing the need for novel tools to improve risk stratification in the elderly. Coronary artery calcification (CAC) is a robust predictor of adverse cardiovascular events, but its long-term prognostic utility beyond RFs in elderly persons is unknown. A consecutive series of 9715 individuals underwent CAC scoring and were followed for a mean of 14.6 ± 1.1 years. Multivariable Cox proportional hazards regression (HR) with 95% confidence intervals (95% CI) was employed to assess the independent relationship of CAC and RFs with all-cause death. The incremental value of CAC, stratified by age, was examined by using an area under the receiver operator characteristic curve (AUC) and category-free net reclassification improvement (NRI). Of the overall study sample, 728 (7.5%) adults (mean age 74.2 ± 4.2 years; 55.6% female) were 70 years or older, of which 157 (21.6%) died. The presence of any CAC was associated with a >4-fold (95% CI = 2.84-6.59) adjusted risk of death for those over the age of 70, which was higher compared with younger study counterparts, or other measured RFs. For individuals 70 years or older, the discriminatory ability of CAC improved upon that of RFs alone (C statistics 0.764 vs. 0.675, P < 0.001). CAC also enabled improved reclassification (category-free NRI = 84%, P < 0.001) when added to RFs. In a large-scale observational cohort registry, CAC improves prediction, discrimination, and reclassification of elderly individuals at risk for future death.


Carbon nanotubes enhance CpG uptake and potentiate antiglioma immunity.
Stimulation of toll-like receptor-9 (TLR9) by CpG oligodeoxynucleotides (CpG) has been shown to counteract the immunosuppressive microenvironment and to inhibit tumor growth in glioma models. Because TLR9 is located intracellularly, we hypothesized that methods that enhance its internalization may also potentiate its immunostimulatory response. The goal of this study was to evaluate carbon nanotubes (CNT) as a CpG delivery vehicle in brain tumor models. Functionalized single-walled CNTs were conjugated with CpG (CNT-CpG) and evaluated in vitro and in mice bearing intracranial GL261 gliomas. Flow cytometry was used to assess CNT-CpG uptake and antiglioma immune response. Tumor growth was measured by bioluminescent imaging, histology, and animal survival. CNT-CpG was nontoxic and enhanced CpG uptake both in vitro and intracranial gliomas. CNT-mediated CpG delivery also potentiated proinflammatory cytokine production by primary monocytes. Interestingly, a single intracranial injection of low-dose CNT-CpG (but not free CpG or blank CNT) eradicated intracranial GL261 gliomas in half of tumor-bearing mice. Moreover, surviving animals exhibited durable tumor-free remission (>3 months), and were protected from intracranial tumor rechallenge, demonstrating induction of long-term antitumor immunity. These findings suggest that CNTs can potentiate CpG immunopotency by enhancing its delivery into tumor-associated inflammatory cells.


Regulation of sigma B levels and activity in Bacillus subtilis.
The sigB operon of Bacillus subtilis encodes sigma B plus three additional proteins (RsbV, RsbW, and RsbX) that regulate sigma B activity. Using an anti-sigma B monoclonal antibody to monitor the levels of sigma B protein, PSPAC to control the expression of the sigB operon, and a ctc-lacZ reporter system to monitor sigma B activity, we observed that the rsbV and rsbW products control sigma B activity at the ctc promoter independently of their effects on sigma B levels. In contrast, RsbX was found to have no effect on expression of ctc when the sigB operon was controlled by PSPAC. The data are consistent with RsbV and RsbW being regulators of sigma B activity and RsbX acting primarily as a negative regulator of sigB operon expression. Evidence that stationary-phase induction of the sigma B-dependent ctc promoter is accomplished by a reduction in RsbW-dependent inhibition of sigma B activity is also presented. In addition, Western blot (immunoblot) analyses of sigB operon expression demonstrated that sigma B accumulation is coupled to the synthesis of its primary inhibitor (RsbW). This finding is consistent with RsbW and sigma B being present within the cell in equivalent amounts, a circumstance that would permit RsbW to directly influence sigma B activity by a direct protein-protein interaction.


Palmitic acid is a toll-like receptor 4 ligand that induces human dendritic cell secretion of IL-1β.
Palmitic acid (PA) and other saturated fatty acids are known to stimulate pro-inflammatory responses in human immune cells via Toll-like receptor 4 (TLR4). However, the molecular mechanism responsible for fatty acid stimulation of TLR4 remains unknown. Here, we demonstrate that PA functions as a ligand for TLR4 on human monocyte derived dendritic cells (MoDCs). Hydrophobicity protein modeling indicated PA can associate with the hydrophobic binding pocket of TLR4 adaptor protein MD-2. Isothermal titration calorimetry quantified heat absorption that occurred during PA titration into TLR4/MD2, indicating that PA binds to TLR4/MD2. Treatment of human MoDCs with PA resulted in endocytosis of TLR4, further supporting the function of PA as a TLR4 agonist. In addition, PA stimulated DC maturation and activation based on the upregulation of DC costimulatory factors CD86 and CD83. Further experiments showed that PA induced TLR4 dependent secretion of the pro-inflammatory cytokine IL-1β. Lastly, our experimental data show that PA stimulation of NF-κB canonical pathway activation is regulated by TLR4 signaling and that reactive oxygen species may be important in upregulating this pro-inflammatory response. Our experiments demonstrate for the first time that PA activation of TLR4 occurs in response to direct molecular interactions between PA and MD-2. In summary, our findings suggest a likely molecular mechanism for PA induction of pro-inflammatory immune responses in human dendritic cells expressing TLR4.


Targeting Human Astrocytes' Calcium-sensing Receptors for Treatment of Alzheimer's Disease.
Understanding the pathophysiology of Alzheimer's disease (AD) in the principal human neural cells is necessary for finding therapeutics for this illness. To help do this, we have been using freshly cultured functionally normal cerebral cortical adult human astrocytes (NAHAs) and postnatal neurons. The findings show that amyloid-β oligomers (Aβ-os) binding to calcium-sensing receptors (CaSRs) on NAHAs and neuron surfaces trigger signals capable of driving AD pathogenesis. This Aβ•CaSR signalling shifts the amyloid precursor protein (APP) from its α-secretase shedding producing neurotrophic/neuroprotective soluble (s)APPα to its β-secretase cleaving engendering AD-driving Aβ42/Aβ42-os peptides. Aβ•CaSR signalling in NAHAs also drives the release of toxic hyper-phosphorylated Tau proteins in exosomes, and of nitric oxide, and VEGF-A. These several harmful agents comprise the neuron-killing machinery, driving the very slowly spreading AD neurocontagion. VEGF-A over-secretion from Aβ-exposed blood vessel-attached astrocytes induces a functional magnetic resonance imaging- detectable hippocampal neoangiogenesis which indicates approaching AD in amnestic minor cognitive impairment (aMCI) patients. Most important in AD's regard, selective allosteric CaSR antagonists (calcylitics) added to Aβ42/Aβ42-os-exposed NAHAs (or to human neuron cultures) rescue the extracellular shedding of neurotrophic/ neuroprotective sAPPα and suppress all the neurotoxic effects of Aβ•CaSR signalling even when multiple microglial cytokines are also present. Therefore, since the multipotent calcilytics would be reasonably safe and inexpensive drugs for humans, it is worthwhile testing them as AD therapeutics in clinical trials especially in persons in the earliest detectable stages of AD neuropathology progression such as aMCI.


Discrimination between chewing of coca leaves or drinking of coca tea and smoking of "paco" (coca paste) by hair analysis. A preliminary study of possibilities and limitations.
Hair analysis is a suitable way to discriminate between coca chewers and consumers of manufactured cocaine using the coca alkaloids hygrine (HYG) and cuscohygrine (CUS) as markers. In the present preliminary study it was examined whether CUS and HYG can be detected in hair of occasional and moderate coca chewers or coca tea drinkers, whether CUS and HYG appear in hair of PACO consumers (smoking coca paste waste), and whether anhydroecgonine methyl ester (AEME) is a useful cocaine smoking marker in this context. Three groups were included: 10 volunteers from Buenos Aires with occasional or moderate chewing of coca leaves or drinking coca tea, 20 Argentinean PACO smokers and 8 German cocaine users. The hair samples (1-4 segments) were analyzed by a validated LC-MS/MS method for cocaine (COC), norcocaine (NC), benzoylecgonine (BE), ecgonine methyl ester (EME), cocaethylene (CE), cinnamoylcocaine (CIN), tropacocaine (TRO), AEME, CUS and HYG. For comparison, eight samples of coca leaves or coca tea were analyzed. Only low concentrations of COC were found in hair of seven occasional users of coca leaves or coca tea (0.010-0.051 ng/mg). For three moderate chewers of coca leaves all compounds were detected including AEME but except TRO. The hair samples of PACO smokers contained much higher concentrations of COC (0.027-341 ng/mg, mean 37.4 ng/mg) and its metabolites. CUS was not found in these samples but traces of HYG were seen in 8 of 37 hair segments. AEME as a marker for coca smoking was detected in hair of 15 smokers. In comparison to COC, the concentrations of EME and CIN were higher for PACO smokers than for German cocaine consumers. AEME (56 ± 20 μg/g) was detected in all coca leave and coca tea samples which explains the detection of this substance in hair of coca chewers. Therefore, its use for differentiation between coca chewers and PACO smokers is limited. CUS remains to be the most suitable marker in hair for chewing coca leaves or drinking coca tea more frequently than two times per month since it does not appear in hair of Argentinean PACO smokers and German cocaine users. Contrary to a previous proposal, the ratios CIN/COC and EME/COC appeared not to be applicable as criteria for this purpose because of the higher concentration of these alkaloids in hair of PACO smokers. More research is needed to assess the value of AEME in hair of South American coca leave or cocaine users.


Cardiomyopathy in zebrafish due to mutation in an alternatively spliced exon of titin.
The zebrafish embryo is transparent and can tolerate absence of blood flow because its oxygen is delivered by diffusion rather than by the cardiovascular system. It is therefore possible to attribute cardiac failure directly to particular genes by ruling out the possibility that it is due to a secondary effect of hypoxia. We focus here on pickwickm171 (pikm171), a recessive lethal mutation discovered in a large-scale genetic screen. There are three other alleles in the pik complementation group with this phenotype (pikm242, pikm740, pikm186; ref. 3) and one allele (pikmVO62H) with additional skeletal paralysis. The pik heart develops normally but is poorly contractile from the first beat. Aside from the edema that inevitably accompanies cardiac dysfunction, development is normal during the first three days. We show by positional cloning that the 'causative' mutation is in an alternatively-spliced exon of the gene (ttn) encoding Titin. Titin is the biggest known protein and spans the half-sarcomere from Z-disc to M-line in heart and skeletal muscle. It has been proposed to provide a scaffold for the assembly of thick and thin filaments and to provide elastic recoil engendered by stretch during diastole. We found that nascent myofibrils form in pik mutants, but normal sarcomeres are absent. Mutant cells transplanted to wildtype hearts remain thin and bulge outwards as individual cell aneurysms without affecting nearby wildtype cardiomyocytes, indicating that the contractile deficiency is cell-autonomous. Absence of Titin function thus results in blockage of sarcomere assembly and causes a functional disorder resembling human dilated cardiomyopathies, one form of which is described in another paper in this issue.


Prefrontal Cortex Activation Reflects Efficient Exploitation of Higher-order Statistical Structure.
Because everyday actions are statistically structured, knowing which action a person has just completed allows predicting the most likely next action step. Taking even more than the preceding action into account improves this predictability but also causes higher processing costs. Using fMRI, we investigated whether observers exploit second-order statistical regularities preferentially if information on possible upcoming actions provided by first-order regularities is insufficient. We hypothesized that anterior pFC balances whether or not second-order information should be exploited. Participants watched videos of actions that were structured by first- and second-order conditional probabilities. Information provided by the first and by the second order was manipulated independently. BOLD activity in the action observation network was more attenuated the more information on upcoming actions was provided by first-order structure, reflecting expectation suppression for more predictable actions. Activation in posterior parietal sites decreased further with second-order information but increased in temporal areas. As expected, second-order information was integrated more when less first-order information was provided, and this interaction was mediated by anterior pFC (BA 10). Observers spontaneously used both the present and the preceding action to predict the upcoming action, and integration of the preceding action was enhanced when the present action was uninformative.


The Marius implant bridge: surgical and prosthetic rehabilitation for the completely edentulous upper jaw with moderate to severe resorption: a 5-year retrospective clinical study.
Patients seeking replacement of their upper denture with an implant-supported restoration are most interested in a fixed restoration. Accompanying the loss of supporting alveolar structure due to resorption is the necessity for lip support, often provided by a denture flange. Attempts to provide a fixed restoration can result in compromises to oral hygiene based on designs with ridge laps. An alternative has been an overdenture prosthesis, which provides lip support but has extensions on to the palate and considerations of patient acceptance. The Marius bridge was developed as a fixed bridge alternative offering lip support that is removable by the patient for hygiene purposes, with no palatal extension beyond normal crown-alveolar contours. Implant-supported restorative treatment of completely edentulous upper jaws, as an alternative to a complete denture, is frequently an elective preference, and it requires significant patient acceptance beyond the functional improvement of chewing. Patients with moderate to severe bone resorption and thin ridges present additional challenges for adequate bone volume and soft-tissue contours. The purpose of this investigation was to develop a surgical and prosthetic implant treatment protocol for completely edentulous maxillae in which optimal lip support and phonetics is achieved in combination with substantial implant anchorage without bone grafting. The Marius bridge is a complete-arch, double-structure prosthesis for maxillae that is removable by the patient for oral hygiene. The first 45 consecutive patients treated by one person (YF) in one center with this concept are reported, with 245 implants followed for up to 5 years after prostheses connection. The cumulative fixture survival rate for this 5-year retrospective clinical study was 97%. Five fixtures failed before loading, in five different patients, and two fixtures in the same patient failed at the 3-year follow-up visit. None of the bridges failed, giving a prostheses survival rate of 100%. The complications were few and mainly prosthetic: nine incidences of attachment component complications, one mesobar fracture, and three reports of gingivitis. All complications were solved or repaired immediately, with minimal or no interruption of prostheses use. Satisfactory medium-term results of survival and patient satisfaction show that the Marius bridge can be recommended for implant dentistry. The technique may reduce the need for grafting, because it allows for longer implants to be placed with improved bone anchorage and prostheses support. key words: clinical follow-up, complete arch, double structure, esthetics, maxilla, phonetics, tilted implants


Estimating the probability of a major outbreak from the timing of early cases: an indeterminate problem?
Conservation biologists, as well as veterinary and public health officials, would benefit greatly from being able to forecast whether outbreaks of infectious disease will be major. For values of the basic reproductive number (R0) between one and two, infectious disease outbreaks have a reasonable chance of either fading out at an early stage or, in the absence of intervention, spreading widely within the population. If it were possible to predict when fadeout was likely to occur, the need for costly precautionary control strategies could be minimized. However, the predictability of even simple epidemic processes remains largely unexplored. Here we conduct an examination of simulated data from the early stages of a fatal disease outbreak and explore how observable information might be useful for predicting major outbreaks. Specifically, would knowing the time of deaths for the first few cases allow us to predict whether an outbreak will be major? Using two approaches, trajectory matching and discriminant function analysis, we find that even in our best-case scenario (with accurate knowledge of epidemiological parameters, and precise times of death), it was not possible to reliably predict the outcome of a stochastic Susceptible-Exposed-Infectious-Recovered (SEIR) process.


Competition Seriousness and Competition Level Modulate Testosterone and Cortisol Responses in Soccer Players.
This study aimed to analyze the modulating effect of competition seriousness and competition level in the testosterone and cortisol responses in professional soccer player. Ninety five (95) soccer players were included in this study (professional, n = 39; semiprofessional, n = 27; amateur, n = 29) before and after training, friendly game and official games. Repeated measures ANOVA showed higher testosterone levels (F(1,89) = 134, p < 0.0001, η2p = 0.75) in professional soccer players, when compared with semiprofessional (p < 0.0001) or amateur athletes (p < 0.0001). After winning a competition game an increase in testosterone levels was observed in professionals (t = -3.456, p < 0.001), semiprofessionals (t = -4.400, p < 0.0001), and amateurs (t = -2.835, p < 0.009). In contrast, this momentary hormonal fluctuation was not observed after winning a friendly game or during a regular training day. Additionally, statistical analysis indicated that cortisol levels were lower in professional (t = -3.456, p < 0.001) and semiprofessional athletes (t = -4.400, p < 0.0001) than in amateurs (t = -2.835, p < 0.009). In soccer players a rise in testosterone was only observable when the team was faced with an actual challenge but did not support a different response between categories. Thus, the desire to achieve a goal (and keep the social status) may be one of the key reasons why testosterone levels rise promptly. Conversely, testosterone did not change after friendly games, which suggests these situations are not real goals and the players do not perceive an actual threat (in terms of dominance) more than the preparation for their next competitive game.