New Mouse Model for Chronic Infections by Gram-Negative Bacteria Enabling the Study of Anti-Infective Efficacy and Host-Microbe Interactions.
Only a few, relatively cumbersome animal models enable long-term Gram-negative bacterial infections that mimic human situations, where untreated infections can last for weeks. Here, we describe a simple murine cutaneous abscess model that enables chronic or progressive infections, depending on the subcutaneously injected bacterial strain. In this model, Pseudomonas aeruginosa cystic fibrosis epidemic isolate LESB58 caused localized high-density skin and soft tissue infections and necrotic skin lesions for up to 10 days but did not disseminate in either CD-1 or C57BL/6 mice. The model was adapted for use with four major Gram-negative nosocomial pathogens, Acinetobacter baumannii, Klebsiella pneumoniae, Enterobacter cloacae, and Escherichia coli This model enabled noninvasive imaging and tracking of lux-tagged bacteria, the influx of activated neutrophils, and production of reactive oxygen-nitrogen species at the infection site. Screening antimicrobials against high-density infections showed that local but not intravenous administration of gentamicin, ciprofloxacin, and meropenem significantly but incompletely reduced bacterial counts and superficial tissue dermonecrosis. Bacterial RNA isolated from the abscess tissue revealed that Pseudomonas genes involved in iron uptake, toxin production, surface lipopolysaccharide regulation, adherence, and lipase production were highly upregulated whereas phenazine production and expression of global activator gacA were downregulated. The model was validated for studying virulence using mutants of more-virulent P. aeruginosa strain PA14. Thus, mutants defective in flagella or motility, type III secretion, or siderophore biosynthesis were noninvasive and suppressed dermal necrosis in mice, while a strain with a mutation in the bfiS gene encoding a sensor kinase showed enhanced invasiveness and mortality in mice compared to controls infected with wild-type P. aeruginosa PA14.IMPORTANCE More than two-thirds of hospital infections are chronic or high-density biofilm infections and difficult to treat due to adaptive, multidrug resistance. Unfortunately, current models of chronic infection are technically challenging and difficult to track without sacrificing animals. Here we describe a model of chronic subcutaneous infection and abscess formation by medically important nosocomial Gram-negative pathogens that is simple and can be used for tracking infections by imaging, examining pathology and immune responses, testing antimicrobial treatments suitable for high-density bacterial infections, and studying virulence. We propose that this mouse model can be a game changer for modeling hard-to-treat Gram-negative bacterial chronic and skin infections.


Collateral Recruitment Is Impaired by Cerebral Small Vessel Disease.
Background and Purpose- Cerebral small vessel disease (SVD) is associated with increased stroke risk and poor stroke outcomes. We aimed to evaluate whether chronic SVD burden is associated with poor recruitment of collaterals in large-vessel occlusive stroke. Methods- Consecutive patients with middle cerebral artery or internal carotid artery occlusion presenting within 6 hours after stroke symptom onset who underwent thrombectomy from 2012 to 2017 were included. The prespecified primary outcome was poor collateral flow, which was assessed on baseline computed tomographic angiography (poor, ≤50% filling; good, >50% filling). Markers of chronic SVD on brain magnetic resonance imaging were rated for the extent of white matter hyperintensities, enlarged perivascular spaces, chronic lacunar infarctions and cerebral microbleeds using the Standards for Reporting Vascular Changes on Neuroimaging criteria. Severity of SVD was quantified by adding the presence of each SVD feature, with a total possible score of 0 to 4; each SVD type was also evaluated separately. Multivariable logistic regression analyses were performed to evaluate the relationships between SVD and poor collaterals, with adjustment for potential confounders. Results- Of the 100 eligible patients, the mean age was 65±16 years, median National Institutes of Health Stroke Scale score was 15, and 68% had any SVD. Poor collaterals were observed in 46%, and those with SVD were more likely to have poor collaterals than patients without SVD (aOR, 1.9 [95% CI, 1.1-3.2]). Of the SVD types, poor collaterals were significantly associated with white matter hyperintensities (aOR, 2.9 per Fazekas increment [95% CI, 1.6-5.3]) but not with enlarged perivascular spaces (adjusted odds ratio [aOR], 1.3 [95% CI, 0.4-4.0]), lacunae (aOR, 2.1 [95% CI, 0.6-7.1]), or cerebral microbleeds (aOR, 2.1 [95% CI, 0.6-7.8]). Having a greater number of different SVD markers was associated with a higher odds of poor collaterals (crude trend P<0.001; adjusted P=0.056). There was a dose-dependent relationship between white matter hyperintensity burden and poor collaterals: adjusted odds of poor collaterals were 1.5, 3.0, and 9.7 across Fazekas scores of 1 to 3 (Ptrend=0.015). No patient with an SVD score of 4 had good collaterals. Conclusions- Chronic cerebral SVD is associated with poor recruitment of collaterals in large vessel occlusive stroke. A prospective study to elucidate the potential mechanism of how SVD may impair the recruitment of collaterals is ongoing.


Revealing Dynamic Rotation of Single Graphene Nanoplatelets on Electrified Microinterfaces.
Nanoparticles interact with a variety of interfaces, from cell walls for medicinal applications to conductive interfaces for energy storage and conversion applications. Unfortunately, quantifying dynamic changes of nanoparticles near interfaces is difficult. While optical techniques exist to study nanoparticle dynamics, motions smaller than the diffraction limit are difficult to quantify. Single-entity electrochemistry has high sensitivity, but the technique suffers from ambiguity in the entity's size, morphology, and collision location. Here, we combine optical microscopy, single-entity electrochemistry, and numerical simulations to elucidate the dynamic motion of graphene nanoplatelets at a gold ultramicroelectrode (radius ∼5 μm). The approach of conductive graphene nanoplatelets, suspended in 10 μM NaOH, to an ultramicroelectrode surface was tracked optically during the continuous oxidation of ferrocenemethanol. Optical microscopy confirmed the nanoplatelet size, morphology, and collision location on the ultramicroelectrode. Nanoplatelets collided on the ultramicroelectrode at an angle, θ, enhancing the electroactive area, resulting in a sharp increase in current. After the collision, the nanoplatelets reoriented to lay flat on the electrode surface, which manifested as a return to the baseline current in the amperometric current-time response. Through correlated finite element simulations, we extracted single nanoplatelet angular velocities on the order of 0.5-2°/ms. These results are a necessary step forward in understanding nanoparticle dynamics at the nanoscale.


Ligand-Stabilized Reduced-Dimensionality Perovskites.
Metal halide perovskites have rapidly advanced thin-film photovoltaic performance; as a result, the materials' observed instabilities urgently require a solution. Using density functional theory (DFT), we show that a low energy of formation, exacerbated in the presence of humidity, explains the propensity of perovskites to decompose back into their precursors. We find, also using DFT, that intercalation of phenylethylammonium between perovskite layers introduces quantitatively appreciable van der Waals interactions. These drive an increased formation energy and should therefore improve material stability. Here we report reduced-dimensionality (quasi-2D) perovskite films that exhibit improved stability while retaining the high performance of conventional three-dimensional perovskites. Continuous tuning of the dimensionality, as assessed using photophysical studies, is achieved by the choice of stoichiometry in materials synthesis. We achieve the first certified hysteresis-free solar power conversion in a planar perovskite solar cell, obtaining a 15.3% certified PCE, and observe greatly improved performance longevity.


Renal involvement in rheumatoid arthritis: analysis of renal biopsy specimens from 100 patients.
Abstract We analyzed renal biopsy specimens from 100 patients to evaluate the characteristics of renal involvement in patients with rheumatoid arthritis (RA). Membranous nephropathy (MN) was the most common renal histological pattern (31%). Mesangial proliferative glomerulonephritis (GN) was found in 21% of cases (IgA nephropathy 12%, non-IgA GN 9%), minor changes in 17%, renal amyloidosis in 11%, interstitial nephritis in 9%, sclerotic GN in 4%, and crescentic GN in 2%. MN was relatively more frequent in men than in women, and most developed nephrotic syndrome, while a few developed renal failure. Disease-modifying antirheumatic drugs (DMARDs) correlated with MN in 26 of 31 cases. Mesangial proliferative GN showed high-grade hematuria. Amyloidosis correlated with long duration of RA; approximately half of the cases with amyloidosis also had nephrotic syndrome, and 82% developed renal failure. Of the 100 patients, 82% showed some tubulo-interstitial changes, which might be related to non-steroidal anti-inflammatory drugs. Because renal lesions in RA are very diverse, and early stage cases of MN and amyloidosis can be detected only by histological examinations, renal biopsy should be performed in cases with continuous urinary abnormalities or progressive renal failure.


Glutamate recognition and hydride transfer by Escherichia coli glutamyl-tRNA reductase.
The initial step of tetrapyrrole biosynthesis in Escherichia coli involves the NADPH-dependent reduction by glutamyl-tRNA reductase (GluTR) of tRNA-bound glutamate to glutamate-1-semialdehyde. We evaluated the contribution of the glutamate moiety of glutamyl-tRNA to substrate specificity in vitro using a range of substrates and enzyme variants. Unexpectedly, we found that tRNA(Glu) mischarged with glutamine was a substrate for purified recombinant GluTR. Similarly unexpectedly, the substitution of amino acid residues involved in glutamate side chain binding (S109A, T49V, R52K) or in stabilizing the arginine 52 glutamate interaction (glutamate 54 and histidine 99) did not abrogate enzyme activity. Replacing glutamine 116 and glutamate 114, involved in glutamate-enzyme interaction near the aminoacyl bond to tRNA(Glu), by leucine and lysine, respectively, however, did abolish reductase activity. We thus propose that the ester bond between glutamate and tRNA(Glu) represents the crucial determinant for substrate recognition by GluTR, whereas the necessity for product release by a 'back door' exit allows for a degree of structural variability in the recognition of the amino acid moiety. Analyzing the esterase activity, which occured in the absence of NADPH, of GluTR variants using the substrate 4-nitrophenyl acetate confirmed the crucial role of cysteine 50 for thioester formation. Finally, the GluTR variant Q116L was observed to lack reductase activity whereas esterase activity was retained. Structure-based molecular modeling indicated that glutamine 116 may be crucial in positioning the nicotinamide group of NADPH to allow for productive hydride transfer to the substrate. Our data thus provide new information about the distinct function of active site residues of GluTR from E. coli.


Structure-guided design of sialic acid-based Siglec inhibitors and crystallographic analysis in complex with sialoadhesin.
The Siglec family of receptors mediates cell surface interactions through recognition of sialylated glycoconjugates. The crystal structure of the N-terminal immunoglobulin-like domain of the Siglec sialoadhesin (SnD1) in complex with 2,3-sialyllactose has informed the design of sialic acid analogs (sialosides) that bind Siglecs with significantly enhanced affinities and specificities. Binding assays against sialoadhesin (Sn; Siglec-1), CD22 (Siglec-2), and MAG (Siglec-4) show a 10- to 300-fold reduction in IC(50) values (relative to methyl-alpha-Neu5Ac) for three sialosides bearing aromatic group modifications of the glycerol side chain: Me-alpha-9-N-benzoyl-amino-9-deoxy-Neu5Ac (BENZ), Me-alpha-9-N-(naphthyl-2-carbonyl)-amino-9-deoxy-Neu5Ac (NAP), and Me-alpha-9-N-(biphenyl-4-carbonyl)-amino-9-deoxy-Neu5Ac (BIP). Crystal structures of these sialosides in complex with SnD1 suggest explanations for the differences in specificity and affinity, providing further ideas for compound design of physiological and potentially therapeutic relevance.


Quantitative proteomics of the tonoplast reveals a role for glycolytic enzymes in salt tolerance.
To examine the role of the tonoplast in plant salt tolerance and identify proteins involved in the regulation of transporters for vacuolar Na(+) sequestration, we exploited a targeted quantitative proteomics approach. Two-dimensional differential in-gel electrophoresis analysis of free flow zonal electrophoresis separated tonoplast fractions from control, and salt-treated Mesembryanthemum crystallinum plants revealed the membrane association of glycolytic enzymes aldolase and enolase, along with subunits of the vacuolar H(+)-ATPase V-ATPase. Protein blot analysis confirmed coordinated salt regulation of these proteins, and chaotrope treatment indicated a strong tonoplast association. Reciprocal coimmunoprecipitation studies revealed that the glycolytic enzymes interacted with the V-ATPase subunit B VHA-B, and aldolase was shown to stimulate V-ATPase activity in vitro by increasing the affinity for ATP. To investigate a physiological role for this association, the Arabidopsis thaliana cytoplasmic enolase mutant, los2, was characterized. These plants were salt sensitive, and there was a specific reduction in enolase abundance in the tonoplast from salt-treated plants. Moreover, tonoplast isolated from mutant plants showed an impaired ability for aldolase stimulation of V-ATPase hydrolytic activity. The association of glycolytic proteins with the tonoplast may not only channel ATP to the V-ATPase, but also directly upregulate H(+)-pump activity.


Reactive species and oxidative stress in optic nerve vulnerable to secondary degeneration.
Secondary degeneration contributes substantially to structural and functional deficits following traumatic injury to the CNS. While it has been proposed that oxidative stress is a feature of secondary degeneration, contributing reactive species and resultant oxidized products have not been clearly identified in vivo. The study is designed to identify contributors to, and consequences of, oxidative stress in a white matter tract vulnerable to secondary degeneration. Partial dorsal transection of the optic nerve (ON) was used to model secondary degeneration in ventral nerve unaffected by the primary injury. Reactive species were assessed using fluorescent labelling and liquid chromatography/tandem mass spectroscopy (LC/MS/MS). Antioxidant enzymes and oxidized products were semi-quantified immunohistochemically. Mitophagy was assessed by electron microscopy. Fluorescent indicators of reactive oxygen and/or nitrogen species increased at 1, 3 and 7days after injury, in ventral ON. LC/MS/MS confirmed increases in reactive species linked to infiltrating microglia/macrophages in dorsal ON. Similarly, immunoreactivity for glutathione peroxidase and haem oxygenase-1 increased in ventral ON at 3 and 7days after injury, respectively. Despite increased antioxidant immunoreactivity, DNA oxidation was evident from 1day, lipid oxidation at 3days, and protein nitration at 7days after injury. Nitrosative and oxidative damage was particularly evident in CC1-positive oligodendrocytes, at times after injury at which structural abnormalities of the Node of Ranvier/paranode complex have been reported. The incidence of mitochondrial autophagic profiles was also significantly increased from 3days. Despite modest increases in antioxidant enzymes, increased reactive species are accompanied by oxidative and nitrosative damage to DNA, lipid and protein, associated with increasing abnormal mitochondria, which together may contribute to the deficits of secondary degeneration.


Mobile phone use and incidence of brain tumour histological types, grading or anatomical location: a population-based ecological study.
Some studies have reported increasing trends in certain brain tumours and a possible link with mobile phone use has been suggested. We examined the incidence time trends of brain tumour in Australia for three distinct time periods to ascertain the influence of improved diagnostic technologies and increase in mobile phone use on the incidence of brain tumours. In a population-based ecological study, we examined trends of brain tumour over the periods 1982-1992, 1993-2002 and 2003-2013. We further compared the observed incidence during the period of substantial mobile phone use (2003-2013) with predicted (modelled) incidence for the same period by applying various relative risks, latency periods and mobile phone use scenarios. National Australian incidence registration data on primary cancers of the brain diagnosed between 1982 and 2013. 16 825 eligible brain cancer cases aged 20-59 from all of Australia (10 083 males and 6742 females). Annual percentage change (APC) in brain tumour incidence based on Poisson regression analysis. The overall brain tumour rates remained stable during all three periods. There was an increase in glioblastoma during 1993-2002 (APC 2.3, 95% CI 0.8 to 3.7) which was likely due to advances in the use of MRI during that period. There were no increases in any brain tumour types, including glioma (-0.6, -1.4 to 0.2) and glioblastoma (0.8, -0.4 to 2.0), during the period of substantial mobile phone use from 2003 to 2013. During that period, there was also no increase in glioma of the temporal lobe (0.5, -1.3 to 2.3), which is the location most exposed when using a mobile phone. Predicted incidence rates were higher than the observed rates for latency periods up to 15 years. In Australia, there has been no increase in any brain tumour histological type or glioma location that can be attributed to mobile phones.


Compensation of signal loss due to cardiac motion in point-resolved spectroscopy of the heart.
To study the influence of cardiac motion on point-resolved single voxel spectroscopy (PRESS) of the human heart and to propose an improved sequence design to compensate for cardiac motion-related signal losses. Numerical simulations and in vivo measurements were performed to study motion effects in cardiac spectroscopy. Based on the findings, an improved PRESS sequence with reduced free induction decay spoiling areas was implemented. Myocardial triglyceride (TG) content as estimated with the improved PRESS sequence was compared with the default PRESS sequence commonly used for spectroscopy in nonmoving tissues that was provided by the manufacturer. Numerical simulations and in vivo data show the importance of optimal trigger delays to avoid signal losses in cardiac spectroscopy. The improved PRESS sequence was found to yield a significantly reduced sensitivity to cardiac motion and trigger delay setting and provided a 1.7-fold increase in signal-to-noise ratio on average compared with the default PRESS sequence. Intrasubject variation of myocardial TG content was reduced with the improved sequence compared with the default PRESS sequence (coefficient of variation: 0.11 ± 0.03 versus 0.23 ± 0.06). Improvement of the sequence design is essential to reduce the sensitivity of PRESS to cardiac motion.


Racial-Ethnic Differences in Turnover Intent Among Home Health Aides.
Home health aides are crucial to ensuring quality services for the growing older adult population needing home-based care in the United States. We utilize data from the nationally representative 2007 National Home Health Aide Survey (N = 3,344) to analyze racial-ethnic disparity in turnover intent. Non-Hispanic Black and non-Hispanic Other home health aides had higher all-cause turnover intent than Non-Hispanic Whites. Cause-specific turnover intent varied by race-ethnicity. Compared with non-Hispanic White home health aides, (a) non-Hispanic Black home health aides were more likely to leave due to low pay and educational/advancement opportunities; (b) Hispanic home health aides were more likely to leave due to a perceived lack of respect or difficulty with their supervisor/agency; and (c) non-Hispanic home health aides were more likely to leave due to an impending move. Findings suggest that efforts to address high levels of turnover intent among home health aides should account for differences in race-ethnic groups.


Vital signs: births to teens aged 15-17 years--United States, 1991-2012.
Teens who give birth at age 15-17 years are at increased risk for adverse medical and social outcomes of teen pregnancy. To examine trends in the rate and proportion of births to teens aged 15-19 years that were to teens aged 15-17 years, CDC analyzed 1991-2012 National Vital Statistics System data. National Survey of Family Growth (NSFG) data from 2006-2010 were used to examine sexual experience, contraceptive use, and receipt of prevention opportunities among female teens aged 15-17 years. During 1991-2012, the rate of births per 1,000 teens declined from 17.9 to 5.4 for teens aged 15 years, 36.9 to 12.9 for those aged 16 years, and 60.6 to 23.7 for those aged 17 years. In 2012, the birth rate per 1,000 teens aged 15-17 years was higher for Hispanics (25.5), non-Hispanic blacks (21.9), and American Indians/Alaska Natives (17.0) compared with non-Hispanic whites (8.4) and Asians/Pacific Islanders (4.1). The rate also varied by state, ranging from 6.2 per 1,000 teens aged 15-17 years in New Hampshire to 29.0 in the District of Columbia. In 2012, there were 86,423 births to teens aged 15-17 years, accounting for 28% of all births to teens aged 15-19 years. This percentage declined from 36% in 1991 to 28% in 2012 (p<0.001). NSFG data for 2006-2010 indicate that although 91% of female teens aged 15-17 years received formal sex education on birth control or how to say no to sex, 24% had not spoken with parents about either topic; among sexually experienced female teens, 83% reported no formal sex education before first sex. Among currently sexually active female teens (those who had sex within 3 months of the survey) aged 15-17 years, 58% used clinical birth control services in the past 12 months, and 92% used contraception at last sex; however, only 1% used the most effective reversible contraceptive methods. Births to teens aged 15-17 years have declined but still account for approximately one quarter of births to teens aged 15-19 years. These data highlight opportunities to increase younger teens exposure to interventions that delay initiation of sex and provide contraceptive services for those who are sexually active; these strategies include support for evidence-based programs that reach youths before they initiate sex, resources for parents in talking to teens about sex and contraception, and access to reproductive health-care services.


Cost-benefit analysis for invasive species control: the case of greater Canada goose Branta canadensis in Flanders (northern Belgium).
Sound decisions on control actions for established invasive alien species (IAS) require information on ecological as well as socio-economic impact of the species and of its management. Cost-benefit analysis provides part of this information, yet has received relatively little attention in the scientific literature on IAS. We apply a bio-economic model in a cost-benefit analysis framework to greater Canada goose Branta canadensis, an IAS with documented social, economic and ecological impacts in Flanders (northern Belgium). We compared a business as usual (BAU) scenario which involved non-coordinated hunting and egg destruction with an enhanced scenario based on a continuation of these activities but supplemented with coordinated capture of moulting birds. To assess population growth under the BAU scenario we fitted a logistic growth model to the observed pre-moult capture population. Projected damage costs included water eutrophication and damage to cultivated grasslands and were calculated for all scenarios. Management costs of the moult captures were based on a representative average of the actual cost of planning and executing moult captures. Comparing the scenarios with different capture rates, different costs for eutrophication and various discount rates, showed avoided damage costs were in the range of 21.15 M€ to 45.82 M€ under the moult capture scenario. The lowest value for the avoided costs applied to the scenario where we lowered the capture rate by 10%. The highest value occurred in the scenario where we lowered the real discount rate from 4% to 2.5%. The reduction in damage costs always outweighed the additional management costs of moult captures. Therefore, additional coordinated moult captures could be applied to limit the negative economic impact of greater Canada goose at a regional scale. We further discuss the strengths and weaknesses of our approach and its potential application to other IAS.


The SAM domain of human TEL2 can abrogate transcriptional output from TEL1 (ETV-6) and ETS1/ETS2.
Regulation of gene expression downstream of the Receptor Tyrosine Kinase signaling pathway in Drosophila relies on a transcriptional effector network featuring two conserved Ets family proteins, Yan and Pointed, known as TEL1 (ETV6) and ETS1/ETS2, respectively, in mammals. As in Drosophila, both TEL1 and ETS1/ETS2 operate as Ras pathway transcriptional effectors and misregulated activity of either factor has been implicated in many human leukemias and solid tumors. Providing essential regulation to the Drosophila network, direct interactions with the SAM domain protein Mae attenuate both Yan-mediated repression and PointedP2-mediated transcriptional activation. Given the critical contributions of Mae to the Drosophila circuitry, we investigated whether the human Ets factors TEL1 and ETS1/ETS2 could be subject to analogous regulation. Here we demonstrate that the SAM domain of human TEL2 can inhibit the transcriptional activities of ETS1/2 and TEL1. Drosophila Mae can also attenuate human ETS1/ETS2 function, suggesting there could be cross-species conservation of underlying mechanism. In contrast, Mae is not an effective inhibitor of TEL1, suggesting the mode of TEL2SAM-mediated inhibition of TEL1 may be distinct from how Drosophila Mae antagonizes Yan. Together our results reveal both further similarities and new differences between the mammalian and Drosophila networks and more broadly suggest that SAM domain-mediated interactions could provide an effective mechanism for modulating output from the TEL1 and ETS1/2 oncogenes.


Illness Perception and Psychological Distress in Persons with Porphyria Cutanea Tarda.
Porphyria cutanea tarda (PCT) requires long-term treatment and follow-up, although many patients experience life-long remission. The aim of this cross-sectional postal survey was to describe and investigate the association between illness perception, health complaints, self-reported symptoms and distress in persons with PCT. The participants perceived PCT as a chronic condition with high levels of personal and treatment control. Persons who reported active symptoms scored higher on perceived illness threat, total health complaints and psychological distress compared with those in remission or latent phases. However, a higher perception of illness threat and the total burden of health complaints were more closely associated with psychological distress than were perceived PCT symptoms activity. This has implications for clinical consultation; dermatologists should be attentive to symptoms activity, but also recognize that patients in remission with a high perceived illness threat and multiple health complaints might be especially vulnerable to psychological distress with regards to PCT.


Within and between whorls: comparative transcriptional profiling of Aquilegia and Arabidopsis.
The genus Aquilegia is an emerging model system in plant evolutionary biology predominantly because of its wide variation in floral traits and associated floral ecology. The anatomy of the Aquilegia flower is also very distinct. There are two whorls of petaloid organs, the outer whorl of sepals and the second whorl of petals that form nectar spurs, as well as a recently evolved fifth whorl of staminodia inserted between stamens and carpels. We designed an oligonucleotide microarray based on EST sequences from a mixed tissue, normalized cDNA library of an A. formosa x A. pubescens F2 population representing 17,246 unigenes. We then used this array to analyze floral gene expression in late pre-anthesis stage floral organs from a natural A. formosa population. In particular, we tested for gene expression patterns specific to each floral whorl and to combinations of whorls that correspond to traditional and modified ABC model groupings. Similar analyses were performed on gene expression data of Arabidopsis thaliana whorls previously obtained using the Ath1 gene chips (data available through The Arabidopsis Information Resource). Our comparative gene expression analyses suggest that 1) petaloid sepals and petals of A. formosa share gene expression patterns more than either have organ-specific patterns, 2) petals of A. formosa and A. thaliana may be independently derived, 3) staminodia express B and C genes similar to stamens but the staminodium genetic program has also converged on aspects of the carpel program and 4) staminodia have unique up-regulation of regulatory genes and genes that have been implicated with defense against microbial infection and herbivory. Our study also highlights the value of comparative gene expression profiling and the Aquilegia microarray in particular for the study of floral evolution and ecology.


Zoonotic Semiotics: Plague Narratives and Vanishing Signs in Madagascar.
Zoonosis calls for a multispecies approach to medical semiotics, a method involving the decipherment of outward symptoms and the construction of narrative. In Madagascar, early detection of bubonic plague outbreaks relies on sightings of sick and dead rats. However, people most vulnerable to plague often do not perceive warning signs, and plague symptoms do not always present in rat and human bodies. In August 2015, a plague outbreak killed 10 residents of a rural hamlet in the central highlands. To reconstruct the transmission chain, scientists elicited survivors' memories of dead rats in the vicinity. Not only were these clues imperceptible to most, but residents had also constructed an alternative outbreak narrative based on different evidence. Stark health disparities, a lack of historical memory of the plague, and genetic adaptations of rats and plague bacteria have created a problem of "semiotic cluelessness" that complicates outbreak control measures and increases mortality.


Cardiovascular Outcomes Reported in Hemodialysis Trials.
Patients on long-term hemodialysis are at very high risk for cardiovascular disease but are usually excluded from clinical trials conducted in the general population or in at-risk populations. There are no universally agreed cardiovascular outcomes for trials conducted specifically in the hemodialysis population. In this review, we highlight that trials reporting cardiovascular outcomes in hemodialysis patients are usually of short duration (median 3 to 6 months) and are small (59% of trials have <100 participants). Overall, the cardiovascular outcomes are very heterogeneous and may not reflect outcomes that are meaningful to patients and clinicians in supporting decision making, as they are often surrogates of uncertain clinical importance. Composite outcomes used in different trials rarely share the same components. In a field in which a single trial is often insufficiently powered to fully assess the clinical and economic impact of interventions, differences in outcome reporting across trials make the task of meta-analysis and interpretation of all the available evidence challenging. Core outcome sets are now being established across many specialties in health care to prevent these problems. Through the global Standardized Outcomes in Nephrology-Hemodialysis initiative, cardiovascular disease was identified as a critically important core domain to be reported in all trials in hemodialysis. Informed by the current state of reporting of cardiovascular outcomes, a core outcome measure for cardiovascular disease is currently being established with involvement of patients, caregivers, and health professionals. Consistent reporting of cardiovascular outcomes that are critically important to hemodialysis patients and clinicians will strengthen the evidence base to inform care in this very high-risk population.


Hydrogen storage in engineered carbon nanospaces.
It is shown how appropriately engineered nanoporous carbons provide materials for reversible hydrogen storage, based on physisorption, with exceptional storage capacities (approximately 80 g H2/kg carbon, approximately 50 g H2/liter carbon, at 50 bar and 77 K). Nanopores generate high storage capacities (a) by having high surface area to volume ratios, and (b) by hosting deep potential wells through overlapping substrate potentials from opposite pore walls, giving rise to a binding energy nearly twice the binding energy in wide pores. Experimental case studies are presented with surface areas as high as 3100 m(2) g(-1), in which 40% of all surface sites reside in pores of width approximately 0.7 nm and binding energy approximately 9 kJ mol(-1), and 60% of sites in pores of width>1.0 nm and binding energy approximately 5 kJ mol(-1). The findings, including the prevalence of just two distinct binding energies, are in excellent agreement with results from molecular dynamics simulations. It is also shown, from statistical mechanical models, that one can experimentally distinguish between the situation in which molecules do (mobile adsorption) and do not (localized adsorption) move parallel to the surface, how such lateral dynamics affects the hydrogen storage capacity, and how the two situations are controlled by the vibrational frequencies of adsorbed hydrogen molecules parallel and perpendicular to the surface: in the samples presented, adsorption is mobile at 293 K, and localized at 77 K. These findings make a strong case for it being possible to significantly increase hydrogen storage capacities in nanoporous carbons by suitable engineering of the nanopore space.


Dissolution of pyroxenes and amphiboles during weathering.
Augite, hypersthene, diopside, and hornblende all undergo dissolution during weathering by means of the formation, growth, and coalescence of distinctive, parallel, lens-shaped etch pits. Similar etch features can be produced if these minerals are treated in the laboratory with concentrated hydrofluoric acid plus hydrochloric acid. These pits most likely form at dislocation outcrops, and their shape and orientation are controlled primarily by the crystallography of the underlying mineral. The results are similar to those found for soil feldspars and suggest that silicate weathering, in general, takes place by selective etching and not by general attack of the surface with consequent rounding as necessiated by bulk diffusion-type weathering theories.


Contact dependent suppression of CD4 T cell activation and proliferation by B cells activated through IgD cross-linking.
Although the co-stimulatory interaction between B and T cells is well defined, recent evidence suggests that B cells also have a regulatory role. Here, we show that B cells activated using anti-IgD conjugated to dextran (α-δ-dex) directly inhibit TCR-induced CD4 T cell activation, proliferation and cytokine production. This effect was observed in CD4 T cells activated both with and without CD28 co-stimulation. T cell viability was unaffected, and the T cell suppressive effect was mediated by contact with IgD activated purified B cells and not by IL-10 or other soluble factors. This is the first evidence of IgD activated B cells mediating inhibition of activation and proliferation of CD4 T cells in humans. This article is protected by copyright. All rights reserved.


Evaluating the WHO Assessment Instrument for Mental Health Systems by comparing mental health policies in four countries.
Mental health is a low priority in most countries around the world. Minimal research and resources have been invested in mental health at the national level. As a result, WHO has developed the Assessment Instrument for Mental Health Systems (WHO-AIMS) to encourage countries to gather data and to re-evaluate their national mental health policy. This paper demonstrates the utility and limitations of WHO-AIMS by applying the model to four countries with different cultures, political histories and public health policies: Iraq, Japan, the Philippines and The former Yugoslav Republic of Macedonia. WHO-AIMS provides a useful model for analysing six domains: policy and legislative framework; mental health services; mental health in primary care; human resources; education of the public at large; and monitoring and research. This is especially important since most countries do not have experts in mental health policy or resources to design their own evaluation tools for mental health systems. Furthermore, WHO-AIMS provides a standardized database for cross-country comparisons. However, limitations of the instrument include the neglect of the politics of mental health policy development, underestimation of the role of culture in mental health care utilization, and questionable measurement validity.


NTP Carcinogenesis Bioassay of Diallyl Phthalate (CAS No. 131-17-9) in B6C3F1 Mice (Gavage Study).
Diallyl phthalate is a widely used crosslinking agent for unsaturated polyesters. Diallyl phthalate or diallyl phthalate polyester blends are used primarily as plasticizers and carriers for adding catalysts and pigments to polyesters and in molding, electrical parts, laminating compounds, and impregnation of metal castings. Rubber compounds, epoxy formulations, and polyurethane foams may also contain diallyl phthalate. Annual production of diallyl phthalate in the United States exceeds 5,000 pounds; precise figures are not available. A NTP Carcinogenesis bioassay of diallyl phthalate (99% pure) was conducted by administering 0 (vehicle control), 150, or 300 mg/kg diallyl phthalate in corn oil by gavage, 5 days per week for 103 weeks, to groups of 50 male and 50 female B6C3F1 mice. Survival rates and mean body weights of dosed mice were not different from those of the controls, and pathological lesions unrelated to proliferative changes were not observed. Therefore, a maximally tolerated dose for the purposes of carcinogenicity testing may not have been achieved. The incidences of lymphoma and either lymphoma or leukemia in dosed male mice were no significantly greater than those in the controls according to pairwise comparisons (P=0.051 to P=0.096), but the trend tests were statistically significant by either life table or incidental tumor analysis (P=0.031 to P=0.045). The incidence of lymphomas in the high-dose male mice was 12/50 (24%) in comparison with 6/50 (12%) in the controls. Recent historical incidences at the performing laboratory and in the NTP Bioassay Program were 18/120 (15%) and 71/661 (11%), respectively. Since the incidence of high-dose male mice with leukemia was not significantly greater than that of concurrent or historical controls at the performing laboratory by pairwise comparisons, this marginal increase was considered only to be equivocally related to diallyl phthalate administration. Increased incidences of squamous cell papillomas, hyperplasia, and inflammatory lesions of the forestomach were observed in diallyl phthalate-dosed mice of both sexes in a dose-related manner. Papillomas of the forestomach were observed in 0%, 2%, and 4% of the control, low-dose, and high-dose mice of both sexes. The recent historical incidence of this tumor in gavage control mice from both the performing laboratory and other laboratories within the Bioassay Program was less than 1%. Forestomach hyperplasia was diagnosed in 0%, 15%, and 18%, and in 8%, 2%, and 29% of the control, low-dose, and high-dose male and female mice, respectively; chronic inflammation of the forestomach was diagnosed in 0%, 9%, and 16% and in 4%, 2%, and 18% of the control, low-dose, and high-dose male and female mice, respectively. Because of the numerical elevation of the forestomach papillomas in the high-dose mice of both sexes, the concomitant observation of dose related forestomach hyperplasia, and the rarity of this tumor in corn oil (gavage) control B6C3F1 mice, the development of squamous cell papillomas of the forestomach may have been related to diallyl phthalate administration. Under the conditions of this bioassay, the development of chronic inflammation and hyperplasia of the forestomach in both male and female B6C3F1 mice was considered to be related to the administration of diallyl phthalate. The development of squamous cell papillomas of the forestomach may also have been related to chemical administration, but the available data are insufficient to indicate a clear cause and effect relationship. An increase in the incidence of male mice with lymphomas was observed, but this increase was considered only to be equivocally related to diallyl phthalate administration. The results of this bioassay, therefore, do not indicate that diallyl phthalate is carcinogenic in B6C3F1 mice, although a maximum tolerated dose may not have been achieved. A carcinogenicity study by the National Toxicology Program of diallyl phthalate in male and female Fisher 344/N rats, employing daily gavage doses of 0 (vehicle control), 50, or 100 mg/kg boicology Program of diallyl phthalate in male and female Fisher 344/N rats, employing daily gavage doses of 0 (vehicle control), 50, or 100 mg/kg body weight is currently being evaluated. Levels of Evidence of Carcinogenicity: Male Mice: Equivocal Female Mice: Equivocal


Low-noise diode-pumped intracavity-doubled laser with off-axially cut Nd:YVO(4).
Effective noise reduction in a diode-pumped intracavity-doubled Nd:YVO(4) laser is achieved with a novel cavity design in which a birefringent filter is formed with off-axially cut Nd:YVO(4) and KTP. 20-mW green output with relative intensity noise of less than -120 dB/Hz was obtained under 290-mW pumping by a broad-area laser diode.


Use of the therapeutic footwear benefit among diabetic medicare beneficiaries in three states, 1995.
To determine the extent to which Medicare provided reimbursement for therapeutic footwear to diabetic Medicare beneficiaries in Washington, Alaska, and Idaho in 1995. Using inpatient, outpatient, and durable medical equipment claims data, we selected a cohort of diabetic Medicare beneficiaries. Therapeutic footwear claims were identified using a set of billing codes intended only for the diabetes footwear benefit. People at "high risk" or "possibly increased risk" for foot problems who might benefit from therapeutic footwear were identified using a combination of International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM) diagnostic codes in any of the databases. Among 608,804 beneficiaries, 10.2% (62,170) met the inclusion criteria for diabetes. Of the diabetic beneficiaries, 13.0% (8,079) had at least one "high risk" diagnosis, and 14.0% (8,686) had at least one "possibly increased risk" diagnosis. The percentage of diabetic beneficiaries with therapeutic footwear claims was 2.9% among those with diagnoses high risk, 0.7% among those with diagnoses indicating possibly increased risk, and 0.1% among those with no diagnosis from the list. Altogether, only 0.6% of beneficiaries meeting the diabetes case ascertainment criteria had a therapeutic footwear claim in 1995. Few diabetic Medicare beneficiaries in Washington, Alaska, and Idaho had claims for reimbursement for therapeutic footwear in 1995. The low utilization of the footwear benefit may represent an important opportunity to improve care for Medicare beneficiaries with diabetes. Further work should be done to characterize the use of the benefit in other regions and to assess whether the low level of usage reflects underutilization.


Engineering the substrate specificity of ADP-ribosyltransferases for identifying direct protein targets.
Adenosine diphosphate ribosyltransferases (ARTDs; ARTD1-17 in humans) are emerging as critical regulators of cell function in both normal physiology and disease. These enzymes transfer the ADP-ribose moiety from its substrate, nicotinamide adenine dinucleotide (NAD(+)), to amino acids of target proteins. The functional redundancy and overlapping target specificities among the 17 ARTDs in humans make the identification of direct targets of individual ARTD family members in a cellular context a formidable challenge. Here we describe the rational design of orthogonal NAD(+) analogue-engineered ARTD pairs for the identification of direct protein targets of individual ARTDs. Guided by initial inhibitor studies with nicotinamide analogues containing substituents at the C-5 position, we synthesized an orthogonal NAD(+) variant and found that it is used as a substrate for several engineered ARTDs (ARTD1, -2, and -6) but not their wild-type counterparts. Comparing the target profiles of ARTD1 (PARP1) and ARTD2 (PARP2) in nuclear extracts highlighted the semi-complementary, yet distinct, protein targeting. Using affinity purification followed by tandem mass spectrometry, we identified 42 direct ARTD1 targets and 301 direct ARTD2 targets. This represents a powerful new technique for identifying direct protein targets of individual ARTD family members, which will facilitate studies delineating the pathway from ARTD activation to a given cellular response.


Psychometric properties of the Beck Self-Esteem Scales.
The psychometric characteristics of the Beck Self-Esteem Scales (BSE) are described for 360 psychiatric outpatients. Patients rated their evaluative beliefs about themselves (Self Scale) and their beliefs about how others evaluate them (Other Scale). Both measures consist of 18 pairs of adjectives, e.g., lovable-unlovable, that are rated using a 10-point scale. The coefficient alphas for the Self Scale (0.94) and the Other Scale (0.95) indicated high internal consistency. Both scales were positively associated with other measures of self-esteem and negatively associated with measures of anxiety and depression. As predicted by the cognitive theory of depression, patients with a principal mood disorder scored significantly lower on the BSE than patients with a principal anxiety disorder. In addition, the mean scores for patients with major depression and dysthymia on the Self Scale were significantly lower than the mean scores for the Other Scale.


Therapeutic effects of the combination of methotrexate and bucillamine in early rheumatoid arthritis: a multicenter, double-blind, randomized controlled study.
Disease-modifying antirheumatic drug (DMARD) combination therapies are used widely, but there have been few reports clearly demonstrating that combination therapy is more effective than DMARD monotherapy. We conducted a multicenter, double-blind controlled trial in order to clarify that the combination of methotrexate and bucillamine is more effective than either alone. The subjects of this study were 71 patients with active rheumatoid arthritis within 2 years of onset. Dosages were 8 mg methotrexate with 5 mg folic acid per week (MTX group), 200 mg bucillamine per day (BUC group), or both MTX and BUC (combination group). Clinical effects and adverse reactions were observed for 96 weeks. The ACR 20 response rate was 79.2% in the combination group, significantly higher than the rates of 43.5% for the MTX group (P = 0.008) and 45.8% for the BUC group (P = 0.0178). The cumulative survival curve of maintaining the ACR 20 response was significantly higher in the combination group than in the MTX and BUC groups (P = 0.0123 and P = 0.0088, respectively). The mean increase in the total Sharp score over 96 weeks was 12.6 +/- 9.0 in the combination group, significantly lower (P = 0.0468) than the value of 28.0 +/- 28.3 for the single DMARD (combined MTX and BUC) group. The incidence of adverse reactions did not differ significantly between the three groups. It was concluded that the combination therapy with MTX and BUC showed significantly higher clinical efficacy than either of the single DMARD therapies.


Improved prediction of cardiovascular disease based on a panel of single nucleotide polymorphisms identified through genome-wide association studies.
Genome-wide association studies (GWAS) have identified single-nucleotide polymorphisms (SNPs) at multiple loci that are significantly associated with coronary artery disease (CAD) risk. In this study, we sought to determine and compare the predictive capabilities of 9p21.3 alone and a panel of SNPs identified and replicated through GWAS for CAD. We used the Ottawa Heart Genomics Study (OHGS) (3323 cases, 2319 control subjects) and the Wellcome Trust Case Control Consortium (WTCCC) (1926 cases, 2938 control subjects) data sets. We compared the ability of allele counting, logistic regression, and support vector machines. Two sets of SNPs, 9p21.3 alone and a set of 12 SNPs identified by GWAS and through a model-fitting procedure, were considered. Performance was assessed by measuring area under the curve (AUC) for OHGS using 10-fold cross-validation and WTCCC as a replication set. AUC for logistic regression using OHGS increased significantly from 0.555 to 0.608 (P=3.59×10⁻¹⁴) for 9p21.3 versus the 12 SNPs, respectively. This difference remained when traditional risk factors were considered in a subgroup of OHGS (1388 cases, 2038 control subjects), with AUC increasing from 0.804 to 0.809 (P=0.037). The added predictive value over and above the traditional risk factors was not significant for 9p21.3 (AUC 0.801 versus 0.804, P=0.097) but was for the 12 SNPs (AUC 0.801 versus 0.809, P=0.0073). Performance was similar between OHGS and WTCCC. Logistic regression outperformed both support vector machines and allele counting. Using the collective of 12 SNPs confers significantly greater predictive capabilities for CAD than 9p21.3, whether traditional risks are or are not considered. More accurate models probably will evolve as additional CAD-associated SNPs are identified.


Selective area heteroepitaxy of GaSb on GaAs (001) for in-plane InAs nanowire achievement.
The growth of in-plane GaSb nanotemplates on a GaAs (001) substrate is demonstrated combining nanoscale patterning of the substrate and selective area heteroepitaxy. The selective growth of GaSb inside nano-stripe openings in a SiO2 mask layer is achieved at low temperature thanks to the use of an atomic hydrogen flux during the molecular beam epitaxy. These growth conditions promote the spreading of GaSb inside the apertures and lattice mismatch accommodation via the formation of a regular array of misfit dislocations at the interface between GaSb and GaAs. We highlight the impact of the nano-stripe orientation as well as the role of the Sb/Ga flux ratio on the strain relaxation of GaSb along the [110] direction and on the nanowire length along the [1-10] one. Finally we demonstrate how these GaSb nanotemplates can be used as pedestals for subsequent growth of in-plane InAs nanowires.


Tumour associated lymphocytes in the pleural effusions of patients with mesothelioma express high levels of inhibitory receptors.
Pleural effusion (PE) is a common feature of malignant pleural mesothelioma. These effusions typically contain lymphocytes and malignant cells. We postulated that the PE would be a source of lymphocytes for analysis of tumor immune milieu. The aim of this study was to compare the phenotype and T cell receptor usage of pleural effusion T cells with paired concurrently drawn peripheral blood lymphocytes. We used multi-parameter flow cytometry and high-throughput T cell receptor sequencing to analyse peripheral blood and pleural effusion mononuclear cells. Both CD8+ and CD4+ T cells from effusion showed increased expression of T cell inhibitory receptors PD-1, LAG-3 and Tim-3 compared to blood. Comprehensive T cell receptor sequencing on one of the patients showed a discordant distribution of clonotypes in the antigen-experienced (PD-1+) compartment between effusion and blood, suggesting an enrichment of antigen specific clonotypes in the effusion, with potential as an immunological response biomarker.


Efficacy of attractive toxic sugar baits (ATSB) against Aedes albopictus with garlic oil encapsulated in beta-cyclodextrin as the active ingredient.
We tested the efficacy of attractive toxic sugar bait (ATSB) with garlic oil microencapsulated in beta-cyclodextrin as active ingredient against Aedes albopictus in suburban Haifa, Israel. Two three-acre gardens with high numbers of Ae. albopictus were selected for perimeter spray treatment with ATSB and ASB (bait containing no active ingredient). Baits were colored with food dye to verify feeding of the mosquitoes. The mosquito population was monitored by human landing catches and sweep net catches in the surrounding vegetation. Experiments lasted for 44 days. Treatment occurred on day 13. The mosquito population collapsed about 4 days after treatment and continued to drop steadily for 27 days until the end of the study. At the experimental site the average pre-treatment landing rate was 17.2 per 5mins. Two days post-treatment, the landing rate dropped to 11.4, and continued to drop to an average of 2.6 during the following 26 days. During the same period, the control population was stable. Few sugar fed females (8-10%) approached a human bait and anthrone tests showed relatively small amounts of sugar within their crop/gut. Around 60-70 % of males caught near our human bait were sugar positive which may indicate that the males were feeding on sugar for mating related behavior. From the vegetation treated with the toxic bait, we recovered significantly fewer (about 10-14%) males and females stained by ATSB than at the ASB-treated control. This may indicate that the toxic baits alter the resting behavior of the poisoned mosquitoes within the vegetation. Almost no Ae. albopictus females (5.2±1.4) approached human bait after treatment with ATSB. It therefore appears that microencapsulated garlic oil is an effective pesticide against Ae. albopictus when used in an ATSB system.


MRI of rheumatoid arthritis:comparing the outcome measures in rheumatology clinical trials (OMERACT) scoring and volume of synovitis for the assessment of biologic therapy.
The outcome measures in rheumatology clinical trials (OMERACT) scores are the most mature quantitation system for rheumatoid arthritis (RA) on magnetic resonance imaging (MRI). Direct measuring techniques of synovial volume have been reported with good reproducibility, although few reports have demonstrated the changes of these measures in response to treatment. To assess these clinical responses, we evaluated the correlation of the changes of clinical activity score 28-joints disease activity score (DAS28-CRP) with the changes of OMERACT scores and with synovial volume measurements. Eight RA patients who were treated by biologic agents were examined with MRI of the dominant affected wrist and finger joints before and one year after the treatment. The total OMERACT score was reduced from 48.0 to 41.3, and synovial volume was reduced from 15.4 to 8.8 milliliters. Positive correlations were seen between the changes of DAS28-CRP and the changes of OMERACT synovitis score (r=0.27), OMERACT total score (r=0.43) and synovial volume (r=0.30). Limited to synovium assessment, synovial volume showed a better correlation with DAS28-CRP than the OMERACT synovitis score. On the other hand, the OMERACT total score showed a higher correlation with DAS28-CRP than synovial volume, probably because the OMERACT total score includes scores for bone erosion and bone edema as well.


The immediate effects of fitting and tuning solid ankle-foot orthoses in early stroke rehabilitation.
Ankle-foot orthoses are known to have a generally positive effect on gait in stroke, however the specifc type of AFO and the time point at which it is provided are highly variable in the currently available literature. The objective was to determine the immediate spatiotemporal and kinematic effect of custom-made solid ankle-foot orthoses in early stroke rehabilitation, compared to shod walking. Five male and three female participants were recruited to the study (n = 8), with a mean age of 57 (16) years who were 3.5 (3) weeks post-stroke. Each received a custom-made solid ankle-foot orthosis to a predefined set of design criteria and tuned using heel wedges to control the shank inclination angle during shod walking. Repeated spatiotemporal and three-dimensional gait measures were taken pre- and immediately post-intervention. A pre-post-test experimental study. With the solid ankle-foot orthosis, walking velocity increased from 0.22 (0.2) to 0.36 (0.3) m/s (p < 0.05), overall average step length increased from 0.28 (0.1) to 0.37 (0.1) m (p < 0.05), cadence increased from 45 (19) to 56 (19) steps/min (p < 0.05) and step length symmetry ratio increased from 0.65 (0.2) to 0.74 (0.2) (not significant). No clear changes were observed in the joint kinematics of the hip and knee. In our small group of early stroke patients who were fitted with a solid ankle-foot orthosis, immediate significant improvements occurred in walking speed, step length and cadence, when compared to walking with shoes only. This study provides evidence about the immediate effects of custom solid ankle-foot orthoses on gait of early stroke survivors. Ankle-foot orthosis design specifications are fully described for replication. This study suggests that observing global segment orientation may be more useful than joint angles when fitting and tuning ankle-foot orthoses for optimal ankle-foot orthosis/footwear alignment.


Lymphadenectomy in uterine low-grade endometrial stromal sarcoma: an analysis of 19 cases and a literature review.
The role of lymphadenectomy in low-grade endometrial stromal sarcoma (ESS) is controversial. The risk of nodal metastases ranges from 0% to 44%, but data are inconclusive. The objective of this study and of the literature review was to investigate the incidence of nodal involvement in macroscopically early-stage tumors. All consecutive patients with histologically proven uterine low-grade ESS who underwent surgery in our institution were considered eligible for the analysis. Until July 2006, pelvic systematic lymphadenectomy was performed based on the physician's choice, whereas after July 2006, all women with apparent early-stage tumor underwent systematic pelvic nodes dissection. From January 1985 to March 2009, 64 women were surgically treated and 19 underwent bilateral systematic pelvic lymphadenectomy. The median count of nodes removed was 20 (range, 11-30). Nodal involvement was depicted in 3 of 19 women but only in 1 in 16 (5%) women with apparent early-stage disease. Specifically, we did not find nodal metastases in 9 women with macroscopically early-stage tumors treated after July 2006. According to the literature review, the risk of nodal metastases is 5% (range, 0%-16%) in early-stage disease. There is no indication to offer a systematic lymphadenectomy in apparent early-stage low-grade ESS according to our analysis and to the literature review. Retroperitoneal surgery should be limited in case of bulky nodal disease or in case of nodal recurrences.


Modulation of PLAGL2 transactivation by positive cofactor 2 (PC2), a component of the ARC/Mediator complex.
The pleomorphic adenoma gene (PLAG) family of transcription factors regulates a wide range of physiological processes, including cell proliferation, tissue-specific gene regulation, and embryonic development, although little is known regarding the mechanisms that regulate PLAG protein activity. In this study, a yeast two-hybrid screen identified PC2, a component of the Mediator complex, as a PLAGL2-binding protein. We show that PC2 cooperates with PLAGL2 and PU.1 to enhance the activity of a known PLAGL2 target promoter (NCF2). The PLAGL2-binding element in the NCF2 promoter consisted of the core sequence of the bipartite PLAG1 consensus site, but lacked the G-cluster motif, and was recognized by PLAGL2 zinc fingers 5 and 6. Promoter and PLAGL2 mutants showed that PLAGL2 and PU.1 were required to bind to their respective sites in the promoter, and PC2 knockdown demonstrated that PC2 was essential for enhanced promoter activity. Co-immunoprecipitation and promoter-reporter studies reveal that the effect of PC2 on PLAGL2 target promoter activity was conferred via the C-terminus of PLAGL2, the region that is required for PC2 binding and contains the PLAGL2 activation domain. Importantly, chromatin immunoprecipitation analysis and PC2 knockdown studies confirmed that endogenous PC2 protein associated with the NCF2 promoter in MM1 cells in the region occupied by PLAGL2, and was required for PLAGL2 target promoter activity in TNF-alpha-treated MM1 cells, respectively. Lastly, the expression of another known PLAGL2 target gene, insulin-like growth factor II (IGF-II), was greatly diminished in the presence of PC2 siRNA. Together, the data identify PC2 as a novel PLAGL2-binding protein and important mediator of PLAGL2 transactivation.


Optimising Controlled Human Malaria Infection Studies Using Cryopreserved P. falciparum Parasites Administered by Needle and Syringe.
Controlled human malaria infection (CHMI) studies have become a routine tool to evaluate efficacy of candidate anti-malarial drugs and vaccines. To date, CHMI trials have mostly been conducted using the bite of infected mosquitoes, restricting the number of trial sites that can perform CHMI studies. Aseptic, cryopreserved P. falciparum sporozoites (PfSPZ Challenge) provide a potentially more accurate, reproducible and practical alternative, allowing a known number of sporozoites to be administered simply by injection. We sought to assess the infectivity of PfSPZ Challenge administered in different dosing regimens to malaria-naive healthy adults (n = 18). Six participants received 2,500 sporozoites intradermally (ID), six received 2,500 sporozoites intramuscularly (IM) and six received 25,000 sporozoites IM. Five out of six participants receiving 2,500 sporozoites ID, 3/6 participants receiving 2,500 sporozoites IM and 6/6 participants receiving 25,000 sporozoites IM were successfully infected. The median time to diagnosis was 13.2, 17.8 and 12.7 days for 2,500 sporozoites ID, 2,500 sporozoites IM and 25,000 sporozoites IM respectively (Kaplan Meier method; p = 0.024 log rank test). 2,500 sporozoites ID and 25,000 sporozoites IM have similar infectivities. Given the dose response in infectivity seen with IM administration, further work should evaluate increasing doses of PfSPZ Challenge IM to identify a dosing regimen that reliably infects 100% of participants. ClinicalTrials.gov NCT01465048.


Biophysical mapping of the adenosine A2A receptor.
A new approach to generating information on ligand receptor interactions within the binding pocket of G protein-coupled receptors has been developed, called Biophysical Mapping (BPM). Starting from a stabilized receptor (StaR), minimally engineered for thermostability, additional single mutations are then added at positions that could be involved in small molecule interactions. The StaR and a panel of binding site mutants are captured onto Biacore chips to enable characterization of the binding of small molecule ligands using surface plasmon resonance (SPR) measurement. A matrix of binding data for a set of ligands versus each active site mutation is then generated, providing specific affinity and kinetic information (K(D), k(on), and k(off)) of receptor-ligand interactions. This data set, in combination with molecular modeling and docking, is used to map the small molecule binding site for each class of compounds. Taken together, the many constraints provided by these data identify key protein-ligand interactions and allow the shape of the site to be refined to produce a high quality three-dimensional picture of ligand binding, thereby facilitating structure based drug design. Results of biophysical mapping of the adenosine A(2A) receptor are presented.


Web-based analysis of the mouse transcriptome using Genevestigator.
Gene function analysis often requires a complex and laborious sequence of laboratory and computer-based experiments. Choosing an effective experimental design generally results from hypotheses derived from prior knowledge or experimentation. Knowledge obtained from meta-analyzing compendia of expression data with annotation libraries can provide significant clues in understanding gene and network function, resulting in better hypotheses that can be tested in the laboratory. Genevestigator is a microarray database and analysis system allowing context-driven queries. Simple but powerful tools allow biologists with little computational background to retrieve information about when, where and how genes are expressed. We manually curated and quality-controlled 3110 mouse Affymetrix arrays from public repositories. Data queries can be run against an annotation library comprising 160 anatomy categories, 12 developmental stage groups, 80 stimuli, and 182 genetic backgrounds or modifications. The quality of results obtained through Genevestigator is illustrated by a number of biological scenarios that are substantiated by other types of experimentation in the literature. The Genevestigator-Mouse database effectively provides biologically meaningful results and can be accessed at https://www.genevestigator.ethz.ch.


[IgA vasculitis with massive hemorrhage from the jejunum after steroid administration].
A 45-year-old Japanese man presenting with leg purpura, abdominal pain, and arthralgia was diagnosed with IgA vasculitis. His symptoms resolved after the intravenous administration of prednisolone. However, on day 20 of admission, he experienced bloody discharge and hypovolemic shock. The bleeding point was not identified on contrast-enhanced computed tomography scanning. The blood loss was approximately 10800ml and the patient received transfusions of 48 units of concentrated red blood cells, 18 units of fresh frozen plasma, and 30 units of concentrated platelets. Laparotomy and enteroscopy were performed through the incision of the jejunum to detect the bleeding source. Spurting bleeding was observed during the enteroscopy and partial resection of the jejunum was performed. Histopathological examination of the resected specimen revealed large vessels beneath the jejunal ulcer scar, suggesting bleeding from a Dieulafoy's lesion. Leukocytoclastic vasculitis or cytomegalovirus infection was not observed in the resected specimen. Gastrointestinal symptoms in patients with IgA vasculitis usually improve with bowel rest and conservative treatment. Administration of steroids or factor XIII is recommended for patients with severe abdominal pain refractory to conservative management. Rarely, massive bleeding, perforation, intussusception, and/or intestinal obstruction occur in the gastrointestinal tract and these complications affect patients' prognoses. The clinical course in the present patient indicated that severe bleeding from the gastrointestinal tract can occur even after symptom remission in patients with IgA vasculitis. In such cases, prompt treatment, including laparotomy and/or enteroscopy, is essential.


Phylogeography of 27,000 SARS-CoV-2 Genomes: Europe as the Major Source of the COVID-19 Pandemic.
The novel coronavirus SARS-CoV-2 emerged from a zoonotic transmission in China towards the end of 2019, rapidly leading to a global pandemic on a scale not seen for a century. In order to cast fresh light on the spread of the virus and on the effectiveness of the containment measures adopted globally, we used 26,869 SARS-CoV-2 genomes to build a phylogeny with 20,247 mutation events and adopted a phylogeographic approach. We confirmed that the phylogeny pinpoints China as the origin of the pandemic with major founders worldwide, mainly during January 2020. However, a single specific East Asian founder underwent massive radiation in Europe and became the main actor of the subsequent spread worldwide during March 2020. This lineage accounts for the great majority of cases detected globally and even spread back to the source in East Asia. Despite an East Asian source, therefore, the global pandemic was mainly fueled by its expansion across and out of Europe. It seems likely that travel bans established throughout the world in the second half of March helped to decrease the number of intercontinental exchanges, particularly from mainland China, but were less effective between Europe and North America where exchanges in both directions are visible up to April, long after bans were imposed.


Phenotypical Characteristics of POC1B-Associated Retinopathy in Japanese Cohort: Cone Dystrophy With Normal Funduscopic Appearance.
Cone/cone-rod dystrophy is a large group of retinal disorders with both phonotypic and genetic heterogeneity. The purpose of this study was to characterize the phenotype of eight patients from seven families harboring POC1B mutations in a cohort of the Japan Eye Genetics Consortium (JEGC). Whole-exome sequencing with targeted analyses identified homozygous or compound heterozygous mutations of the POC1B gene in 7 of 548 families in the JEGC database. Ophthalmologic examinations including the best-corrected visual acuity, perimetry, fundus photography, fundus autofluorescence imaging, optical coherence tomography, and full-field and multifocal electroretinography (ERGs) were performed. There were four men and four women whose median age at the onset of symptoms was 15.6 years (range, 6-23 years) and that at the time of examination was 40.3 years (range, 22-67 years). The best-corrected visual acuity ranged from -0.08 to 1.52 logMAR units. The funduscopic appearance was normal in all the cases except in one case with faint mottling in the fovea. Optical coherence tomography revealed an absence of the interdigitation zone and blurred ellipsoid zone in the posterior pole, but the foveal structures were preserved in three cases. The full-field photopic ERGs were reduced or extinguished with normal scotopic responses. The central responses of the multifocal ERGs were preserved in two cases. The diagnosis was either generalized cone dystrophy in five cases or cone dystrophy with foveal sparing in three cases. Generalized or peripheral cone dystrophy with normal funduscopic appearance is the representative phenotype of POC1B-associated retinopathy in our cohort.


A model for the sequential dominance of antigenic variants in African trypanosome infections.
Trypanosoma brucei infects various domestic and wild mammals in equatorial Africa. The parasite's genome contains several hundred alternative and highly diverged surface antigens, of which only a single one is expressed in any cell. Individual cells occasionally change expression of their surface antigen, allowing them to escape immune surveillance. These switches appear to occur in a partly random way, creating a diverse set of antigenic variants. In spite of this diversity, the parasitaemia develops as a series of outbreaks, each outbreak dominated by relatively few antigenic types. Host-specific immunity eventually clears the dominant antigenic types and a new outbreak follows from antigenic types that have apparently been present all along at low frequency. This pattern of sequential dominance by different antigenic types remains unexplained. I use a mathematical model of parasitaemia and host immunity to show that small variations in the rate at which each type switches to other types can explain the observations. My model shows that randomly chosen switch rates do not provide sufficiently ordered parasitaemias to match the observations. Instead, minor modifications of switch rates by natural selection are required to develop a sequence of ordered parasitaemias.


Toxicity of radiotherapy in patients with collagen vascular disease.
A diagnosis of collagen vascular disease (CVD) may predispose to radiotherapy (RT) toxicity. The objective of the current study was to identify factors that influence RT toxicity in the setting of CVD. A total of 86 RT courses for 73 patients with CVD were delivered between 1985 and 2005. CVD subtypes include rheumatoid arthritis (RA; 33 patients), systemic lupus erythematosus (SLE; 13 patients), scleroderma (9 patients), dermatomyositis/polymyositis (5 patients), ankylosing spondylitis (4 patients), polymyalgia rheumatica/temporal arteritis (4 patients), Wegener granulomatosis (3 patients), and mixed connective tissue disorders (MCTD)/other (2 patients). Each patient with CVD was matched to 1 to 3 controls with respect to sex, race, site irradiated, RT dose (+/-2 Gray), and age (+/-5 years). There was no significant difference between CVD patients (65.1%) and controls (72.5%) experiencing any acute toxicity. CVD patients had a higher incidence of any late toxicity (29.1% vs 14%; P = .001), and a trend toward an increased rate of severe late toxicity (9.3% vs 3.7%; P = .079). RT delivered to the breast had increased risk of severe acute toxicity, whereas RT to the pelvis had increased risk of severe acute and late toxicity. RT administered in the setting of scleroderma carried a higher risk of severe late toxicity, whereas RT to SLE patients carried a higher risk of severe acute and late toxicity. Although generally well tolerated, RT in the setting of CVD appears to carry a higher risk of late toxicity. RT to the pelvis or in the setting of SLE or scleroderma may predispose to an even greater risk of severe toxicity. These issues should be considered when deciding whether to offer RT for these patients.


Genetic evidence for spatio-temporal changes in the dispersal patterns of two sympatric African colobine monkeys.
Western black-and-white colobus and Temmink's red colobus are two forest-dependent African primates with similar ecological requirements, often found in sympatry. Their most striking difference lies in their social system: black-and-white colobus live in small groups with mainly male-mediated dispersal but where females can also disperse, whereas red colobus live in larger groups with males described as philopatric. To investigate whether genetic evidence supports the reported patterns of dispersal based on observational data, we examined eight black-and-white and six red colobus social groups from Cantanhez National Park, Guinea-Bissau. Microsatellite markers revealed a lack of sex-biased dispersal for black-and-white colobus. Gene flow, mainly mediated by females, better explained the genetic patterns found in red colobus, with some evidence for less extensive male dispersal. In contrast to the microsatellite data, low mitochondrial diversity for the black-and-white colobus suggests that historical and/or long-range male-mediated gene flow might have been favored. In red colobus, the co-existence of three divergent mitochondrial haplogroups suggests that the Cantanhez population contains a secondary contact zone between divergent lineages that evolved in allopatry. Female-biased dispersal in this species may be a major factor contributing to the colonization by such differentiated mitochondrial lineages in the region. Overall, we find evidence for a spatio-temporal change in the dispersal patterns of the colobus monkeys of Cantanhez, with mitochondrial DNA indicating dispersal by mainly a single sex and microsatellite data suggesting that recently both sexes appear to be dispersing within the population.


Multiple clinical forms of dehydrated hereditary stomatocytosis arise from mutations in PIEZO1.
Autosomal dominant dehydrated hereditary stomatocytosis (DHSt) usually presents as a compensated hemolytic anemia with macrocytosis and abnormally shaped red blood cells (RBCs). DHSt is part of a pleiotropic syndrome that may also exhibit pseudohyperkalemia and perinatal edema. We identified PIEZO1 as the disease gene for pleiotropic DHSt in a large kindred by exome sequencing analysis within the previously mapped 16q23-q24 interval. In 26 affected individuals among 7 multigenerational DHSt families with the pleiotropic syndrome, 11 heterozygous PIEZO1 missense mutations cosegregated with disease. PIEZO1 is expressed in the plasma membranes of RBCs and its messenger RNA, and protein levels increase during in vitro erythroid differentiation of CD34(+) cells. PIEZO1 is also expressed in liver and bone marrow during human and mouse development. We suggest for the first time a correlation between a PIEZO1 mutation and perinatal edema. DHSt patient red cells with the R2456H mutation exhibit increased ion-channel activity. Functional studies of PIEZO1 mutant R2488Q expressed in Xenopus oocytes demonstrated changes in ion-channel activity consistent with the altered cation content of DHSt patient red cells. Our findings provide direct evidence that R2456H and R2488Q mutations in PIEZO1 alter mechanosensitive channel regulation, leading to increased cation transport in erythroid cells.


Impact of body mass index on adjacent segment disease after lumbar fusion for degenerative spine disease.
Adjacent segment disease is an important complication after fusion of degenerative lumbar spines. However, the role of body mass index (BMI) in adjacent segment disease has been addressed less. To examine the relationship between BMI and adjacent segment disease after lumbar fusion for degenerative spine diseases. For this retrospective study, we enrolled 190 patients undergoing lumbar fusion surgery for degeneration. BMI at admission was documented. Adjacent segment disease was defined by integration of the clinical presentations and radiographic criteria based on the morphology of the dural sac on magnetic resonance images. Adjacent segment disease was identified in 13 of the 190 patients, accounting for 6.8%. The interval between surgery and diagnosis as adjacent segment disease ranged from 21 to 66 months. Five of the 13 patients required subsequent surgical intervention for clinically relevant adjacent segment disease. In the logistic regression model, BMI was a risk factor for adjacent segment disease after lumbar fusion for degenerative spine diseases (odds ratio, 1.68; 95% confidence interval, 1.27-2.21; P < .001). Any increase of 1 mean value in BMI would increase the adjacent segment disease rate by 67.6%. The patients were subdivided into 2 groups based on BMI, and up to 11.9% of patients with BMI ≥ 25 kg/m were diagnosed as having adjacent segment disease at the last follow-up. BMI is a risk factor for adjacent segment disease in patients undergoing lumbar fusion for degenerative spine diseases. Because BMI is clinically objective and modifiable, controlling body weight before or after surgery may provide opportunities to reduce the rate of adjacent segment disease and to improve the outcome of fusion surgery.


Laser ablation tomography for visualization of root colonization by edaphic organisms.
Soil biota have important effects on crop productivity, but can be difficult to study in situ. Laser ablation tomography (LAT) is a novel method that allows for rapid, three-dimensional quantitative and qualitative analysis of root anatomy, providing new opportunities to investigate interactions between roots and edaphic organisms. LAT was used for analysis of maize roots colonized by arbuscular mycorrhizal fungi, maize roots herbivorized by western corn rootworm, barley roots parasitized by cereal cyst nematode, and common bean roots damaged by Fusarium. UV excitation of root tissues affected by edaphic organisms resulted in differential autofluorescence emission, facilitating the classification of tissues and anatomical features. Samples were spatially resolved in three dimensions, enabling quantification of the volume and distribution of fungal colonization, western corn rootworm damage, nematode feeding sites, tissue compromised by Fusarium, and as well as root anatomical phenotypes. Owing to its capability for high-throughput sample imaging, LAT serves as an excellent tool to conduct large, quantitative screens to characterize genetic control of root anatomy and interactions with edaphic organisms. Additionally, this technology improves interpretation of root-organism interactions in relatively large, opaque root segments, providing opportunities for novel research investigating the effects of root anatomical phenes on associations with edaphic organisms.


Effects of fixed labial orthodontic appliances on speech sound production.
To explore the impact of fixed labial orthodontic appliances on speech sound production. Speech evaluations were performed on 23 patients with fixed labial appliances. Evaluations were performed immediately prior to appliance insertion, immediately following insertion, and 1 and 2 months post insertion. Baseline dental/skeletal variables were correlated with the ability to accommodate the presence of the appliances. Appliance effects were variable: 44% of the subjects were unaffected, 39% were temporarily affected but adapted within 2 months, and 17% of patients showed persistent sound errors at 2 months. Resolution of acquired sound errors was noted by 8 months post-appliance removal. Maladaptation to appliances was correlated to severity of malocclusion as determined by the Grainger's Treatment Priority Index. Sibilant sounds, most notably /s/, were affected most often. (1) Insertion of fixed labial appliances has an effect on speech sound production. (2) Sibilant and stopped sounds are affected, with /s/ being affected most often. (3) Accommodation to fixed appliances depends on the severity of malocclusion.


MtLAX2, a Functional Homologue of the Arabidopsis Auxin Influx Transporter AUX1, Is Required for Nodule Organogenesis.
Most legume plants can form nodules, specialized lateral organs that form on roots, and house nitrogen-fixing bacteria collectively called rhizobia. The uptake of the phytohormone auxin into cells is known to be crucial for development of lateral roots. To test the role of auxin influx in nodulation we used the auxin influx inhibitors 1-naphthoxyacetic acid (1-NOA) and 2-NOA, which we found reduced nodulation of Medicago truncatula. This suggested the possible involvement of the AUX/LAX family of auxin influx transporters in nodulation. Gene expression studies identified MtLAX2, a paralogue of Arabidopsis (Arabidopsis thaliana) AUX1, as being induced at early stages of nodule development. MtLAX2 is expressed in nodule primordia, the vasculature of developing nodules, and at the apex of mature nodules. The MtLAX2 promoter contains several auxin response elements, and treatment with indole-acetic acid strongly induces MtLAX2 expression in roots. mtlax2 mutants displayed root phenotypes similar to Arabidopsis aux1 mutants, including altered root gravitropism, fewer lateral roots, shorter root hairs, and auxin resistance. In addition, the activity of the synthetic DR5-GUS auxin reporter was strongly reduced in mtlax2 roots. Following inoculation with rhizobia, mtlax2 roots developed fewer nodules, had decreased DR5-GUS activity associated with infection sites, and had decreased expression of the early auxin responsive gene ARF16a Our data indicate that MtLAX2 is a functional analog of Arabidopsis AUX1 and is required for the accumulation of auxin during nodule formation in tissues underlying sites of rhizobial infection.


Age and gender-related differences in emotional and behavioural problems and autistic features in children and adolescents with Down syndrome: a survey-based study of 674 individuals.
Recent studies have indicated an increased risk of autism, behavioural and emotional problems and attention-deficit/hyperactivity disorder in individuals with Down syndrome. In a large-scale survey-based study, we examined the rates of these problems and their relationship to age and gender, in a sample of 674 individuals (4-18 years) with Down syndrome. The relationship with IQ level was also explored in a subsample (n = 175). The Strengths and Difficulties Questionnaire and the Social Communication Questionnaire were used to assess behavioural and emotional problems and autism traits. On the Strengths and Difficulties Questionnaire, peer problems were the most frequently reported difficulty (48% > cut-off), followed by hyperactivity/inattention (34% > cut-off). On the Social Communication Questionnaire, 37% scored at or above cut-off (≥15) for autism spectrum disorder; 17% were at or above the suggested cut-off (≥22) for autism. Little association between age and behavioural or emotional problems or with severity of autistic symptomatology was found. However, peer problems were more common in adolescents than in junior school children (P < 0.001); Hyperactivity/inattention was less prevalent among adolescents (P < 0.001). High rates of autistic features, emotional and behavioural problems are documented. These problems are related to age, gender and degree of intellectual disability.


Accuracy and confidence of visual short-term memory do not go hand-in-hand: behavioral and neural dissociations.
Currently influential models of working memory posit that memory content is highly accessible to conscious inspection. These models predict that metacognition of memory performance should go hand-in-hand with the accuracy of the underlying memory representation. To test this view, we investigated how visual information presented during the maintenance period affects VSTM accuracy and confidence. We used a delayed cue-target orientation discrimination task in which participants were asked to hold in memory a grating, and during the maintenance period a second memory cue could be presented. VSTM accuracy of the first memory cue was impaired when the orientation of the second memory cue was sufficiently different. However, participants' response confidence was reduced whenever the second memory cue was presented; thus VSTM accuracy and confidence were dissociated. In a second experiment, we applied transcranial direct current stimulation (tDCS) over the right dorsolateral prefrontal cortex (DLPFC) to investigate the causal role of this region in VSTM metacognition. Relative to the sham condition, anodal tDCS induced a general reduction in confidence ratings but did not affect VSTM accuracy. Overall, these results indicate that our metacognition of memory performance is influenced by factors other than the accuracy of the underlying memory representation.


Associations Between the Cyclic Guanosine Monophosphate Pathway and Cardiovascular Risk Factors: MESA.
Background cGMP mediates numerous cardioprotective functions and is a potential therapeutic target for cardiovascular disease. Preclinical studies suggest that plasma cGMP is reflective of natriuretic peptide stimulation. Epidemiologic associations between cGMP and natriuretic peptide, as well as cardiovascular disease risk factors, are unknown. Methods and Results We measured plasma cGMP in 542 men and 496 women free of cardiovascular disease and heart failure in MESA (Multi-Ethnic Study of Atherosclerosis). Cross-sectional associations of N-terminal pro-B type natriuretic peptide, sex hormones, and cardiovascular disease/heart failure risk factors with log(cGMP) were analyzed using multivariable linear regression models. Mean (SD) cGMP was 4.7 (2.6) pmol/mL, with no difference between the sexes. After adjusting for cardiovascular risk factors, N-terminal pro-B type natriuretic peptide was significantly positively associated with cGMP (P<0.05). Higher blood pressure and lower estimated glomerular filtration rate were associated with higher cGMP (P<0.05). Triglyceride levels, total/high-density lipoprotein cholesterol ratio, presence of diabetes mellitus, and the homeostatic model assessment of insulin resistance were inversely associated with cGMP (P<0.05). Among women, free testosterone and dehydroepiandrosterone were inversely associated with cGMP, while sex hormone binding globulin was positively associated (P<0.05). Conclusions In a community-cohort, plasma cGMP was associated with natriuretic peptide signaling. Higher blood pressure and greater renal dysfunction were positively associated with cGMP, while adverse metabolic risk factors were inversely associated. Increased androgenicity in postmenopausal women was inversely associated with cGMP. These novel associations further our understanding of the role of cGMP in a general population.


Somatosensory Evoked Potentials Reveal Reduced Embodiment of Emotions in Autism.
Consistent with current models of embodied emotions, this study investigates whether the somatosensory system shows reduced sensitivity to facial emotional expressions in autistic compared with neurotypical individuals, and whether these differences are independent from between-group differences in visual processing of facial stimuli. To investigate the dynamics of somatosensory activity over and above visual carryover effects, we recorded EEG activity from two groups of autism spectrum disorder (ASD) or typically developing (TD) humans (male and female), while they were performing a facial emotion discrimination task and a control gender task. To probe the state of the somatosensory system during face processing, in 50% of trials we evoked somatosensory activity by delivering task-irrelevant tactile taps on participants' index finger, 105 ms after visual stimulus onset. Importantly, we isolated somatosensory from concurrent visual activity by subtracting visual responses from activity evoked by somatosensory and visual stimuli. Results revealed significant task-dependent group differences in mid-latency components of somatosensory evoked potentials (SEPs). ASD participants showed a selective reduction of SEP amplitudes (P100) compared with TD during emotion task; and TD, but not ASD, showed increased somatosensory responses during emotion compared with gender discrimination. Interestingly, autistic traits, but not alexithymia, significantly predicted SEP amplitudes evoked during emotion, but not gender, task. Importantly, we did not observe the same pattern of group differences in visual responses. Our study provides direct evidence of reduced recruitment of the somatosensory system during emotion discrimination in ASD and suggests that this effect is not a byproduct of differences in visual processing.SIGNIFICANCE STATEMENT The somatosensory system is involved in embodiment of visually presented facial expressions of emotion. Despite autism being characterized by difficulties in emotion-related processing, no studies have addressed whether this extends to embodied representations of others' emotions. By dissociating somatosensory activity from visual evoked potentials, we provide the first evidence of reduced recruitment of the somatosensory system during emotion discrimination in autistic participants, independently from differences in visual processing between typically developing and autism spectrum disorder participants. Our study uses a novel methodology to reveal the neural dynamics underlying difficulties in emotion recognition in autism spectrum disorder and provides direct evidence that embodied simulation of others' emotional expressions operates differently in autistic individuals.


Creatine Supplementation and Skeletal Muscle Metabolism for Building Muscle Mass- Review of the Potential Mechanisms of Action.
Creatine, a very popular supplement among athletic populations, is of growing interest for clinical applications. Since over 90% of creatine is stored in skeletal muscle, the effect of creatine supplementation on muscle metabolism is a widely studied area. While numerous studies over the past few decades have shown that creatine supplementation has many favorable effects on skeletal muscle physiology and metabolism, including enhancing muscle mass (growth/hypertrophy); the underlying mechanisms are poorly understood. This report reviews studies addressing the mechanisms of action of creatine supplementation on skeletal muscle growth/hypertrophy. Early research proposed that the osmotic effect of creatine supplementation serves as a cellular stressor (osmosensing) that acts as an anabolic stimulus for protein synthesis signal pathways. Other reports indicated that creatine directly affects muscle protein synthesis via modulations of components in the mammalian target of rapamycin (mTOR) pathway. Creatine may also directly affect the myogenic process (formation of muscle tissue), by altering secretions of myokines, such as myostatin and insulin-like growth factor-1, and expressions of myogenic regulatory factors, resulting in enhanced satellite cells mitotic activities and differentiation into myofiber. Overall, there is still no clear understanding of the mechanisms of action regarding how creatine affects muscle mass/growth, but current evidence suggests it may exert its effects through multiple approaches, with converging impacts on protein synthesis and myogenesis.


Methylmalonic Acidemia Diagnosis by Laboratory Methods.
Methylmalonic acidemia (MMA) is usually caused by a deficiency of the enzyme methylmalonyl-CoA mutase (MCM), a defect in the transport or synthesis of its cofactor, adenosyl-cobalamin (cblA, cblB, cblC, cblF, cblD, and cblX), or deficiency of the enzyme methylmalonyl-CoA epimerase. A comprehensive diagnostic approach involves investigations of metabolites with tandem mass spectrometry, organic acid analysis with gas chromatography, enzymatic studies with fibroblast cell culture, and finally, mutation analysis. With biochemical techniques and enzymatic assay the reliable characterization of patients with isolated MMA for mutation analysis can be achieved. Reliable classification of these patients is essential for ongoing and prospective studies on treatments, outcomes, and prenatal diagnoses. This article reviews the diagnostic techniques used to characterize patients with MMA.


Three dimensional sensitivity characterization of plasmonic nanorods for refractometric biosensors.
An experimental three dimensional characterization of the local refractive index sensitivity of plasmonic gold nanorods is performed by controlled apposition of lithographic nanostructures. We show up to seven times higher sensitivity values to local changes in the refractive index at the particle tip than center. In addition, successive deposition of defined nm-thin dielectric layers on nanorods covered with stripe masks allows us to study the sensitivity decrease normal to the particle surface separately for different particle sites. Clear trends to a stronger sensitivity decay at sites of higher local sensitivity are demonstrated experimentally and theoretically. Our sensitivity characterization provides an important tool to find the most suitable particle type and particle site for specific bio-sensing applications.


Transient chylomicronemia preceding the onset of insulin-dependent diabetes in a young girl with no humoral markers of islet autoimmunity.
We investigated the possible causes of diabetes in a young child who presented with hyperglycemia associated with severe hypertriglyceridemia (>166 mmol/l), hypercholesterolemia (>38 mmol/l) and fasting chilomicrons. The patient did not have any of the HLA and autoantibody markers typically associated with type 1 diabetes. A glucose clamp failed to demonstrate insulin resistance (peripheral glucose utilization rate (M)=4.3 mg/kg per min) and there was no family history of type 2 diabetes or maturity onset diabetes in youth. Both fasting and stimulated C-peptide levels, including those in response to i.v. glucagon, were below the limit of detection. This is consistent with loss of beta-cell function. The family history did not reveal the existence of relatives with lipid abnormalities, coronary heart disease, and pancreatitis. We did not find any abnormality of plasma apoCII, lipoproteinlipase and hepatic lipase activities. The patients had a epsilon3/epsilon3 apoE genotype and she rapidly cleared an oral fat load after normalization of plasma lipids. The mild hyperglycemia seems an unlikely explanation for both the severe hypertriglyceridemia and chylomicronemia. A more plausible explanation is transient lipoproteinlipase deficiency. This rare condition, occasionally associated with a high-fat diet, could have caused the rapid and dramatic hypertriglyceridemia observed in this patient, which in turn might have led to the beta-cell destruction by direct lipid toxicity.


Serial changes in adiponectin and BNP in ACS patients: paradoxical associations with each other and with prognosis.
Plasma adiponectin is inversely associated with the risk of coronary heart disease in healthy people. However, adiponectin and BNP (B-type natriuretic peptide) are both known to be positively associated with a risk of poor outcome, and with each other, in ACS (acute coronary syndrome) patients. Serial changes in plasma adiponectin and BNP following ACS have not been assessed previously, and may clarify these apparently paradoxical associations. In the present study, adiponectin, BNP, classical risk markers and clinical parameters were measured in plasma from 442 consecutive ACS patients in an urban teaching hospital, with repeat measures at 7 weeks (n=338). Patients were followed-up for 10 months. Poor outcome was defined as mortality or readmission for ACS or congestive heart failure (n=90). In unadjusted analysis, the change in adiponectin (but not baseline or 7-week adiponectin) was significantly associated with the risk of an adverse outcome {odds ratio (OR), 5.42 [95% CI (confidence interval), 2.78-10.55]}. This association persisted after adjusting for classical risk factors and clinical markers, but was fully attenuated by adjusting for the 7-week BNP measurement [OR, 1.13 (95% CI, 0.27-4.92)], which itself remained associated with risk [OR, 5.86 (95% CI, 1.04-32.94)]. Adiponectin and BNP positively correlated at baseline and 7 weeks, and the change in both parameters over 7 weeks also correlated (r=0.39, P<0.001). In conclusion, increases in plasma adiponectin (rather than absolute levels) after ACS are related to the risk of an adverse outcome, but this relationship is not independent of BNP levels. The results of the present study allude to a potential direct or indirect relationship between adiponectin and BNP post-ACS which requires further investigation.