Survival functions for defining a clinical management Lost To Follow-Up (LTFU) cut-off in Antiretroviral Therapy (ART) program in Zomba, Malawi.
While, lost to follow-up (LTFU) from antiretroviral therapy (ART) can be considered a catch-all category for patients who miss scheduled visits or medication pick-ups, operational definitions and methods for defining LTFU vary making comparisons across programs challenging. Using weekly cut-offs, we sought to determine the probability that an individual would return to clinic given that they had not yet returned in order to identify the LTFU cut-off that could be used to inform clinical management and tracing procedures. Individuals who initiated ART with Dignitas International supported sites (n = 22) in Zomba, Malawi between January 1 2007-June 30 2010 and were ≥ 1 week late for a follow-up visit were included. Lateness was categorized using weekly cut-offs from ≥1 to ≥26 weeks late. At each weekly cut-off, the proportion of patients who returned for a subsequent follow-up visit were identified. Cumulative Distribution Functions (CDFs) were plotted to determine the probability of returning as a function of lateness. Hazard functions were plotted to demonstrate the proportion of patients who returned each weekly interval relative to those who had yet to return. In total, n = 4484 patients with n = 7316 follow-up visits were included. The number of included follow-up visits per patient ranged from 1-10 (median: 1). Both the CDF and hazard function demonstrated that after being ≥9 weeks late, the proportion of new patients who returned relative to those who had yet to return decreased substantially. We identified a LTFU definition useful for clinical management. The simple functions plotted here did not require advanced statistical expertise and were created using Microsoft Excel, making it a particularly practical method for HIV programs in resource-constrained settings.


Content-specific auditing of a large scale anatomy ontology.
Biomedical ontologies are envisioned to be usable in a range of research and clinical applications. The requirements for such uses include formal consistency, adequacy of coverage, and possibly other domain specific constraints. In this report we describe a case study that illustrates how application specific requirements may be used to identify modeling problems as well as data entry errors in ontology building and evolution. We have begun a project to use the UW Foundational Model of Anatomy (FMA) in a clinical application in radiation therapy planning. This application focuses mainly (but not exclusively) on the representation of the lymphatic system in the FMA, in order to predict the spread of tumor cells to regional metastatic sites. This application requires that the downstream relations associated with lymphatic system components must only be to other lymphatic chains or vessels, must be at the appropriate level of granularity, and that every path through the lymphatic system must terminate at one of the two well known trunks of the lymphatic system. It is possible through a programmable query interface to the FMA to write small programs that systematically audit the FMA for compliance with these constraints. We report on the design of some of these programs, and the results we obtained by applying them to the lymphatic system. The algorithms and approach are generalizable to other network organ systems in the FMA such as arteries and veins. In addition to illustrating exact constraint checking methods, this work illustrates how the details of an application may reflect back a requirement to revise the design of the ontology itself.


Divergent roles of growth factors in the GnRH regulation of puberty in mice.
Pubertal onset, initiated by pulsatile gonadotropin-releasing hormone (GnRH), only occurs in a favorable, anabolic hormonal milieu. Anabolic factors that may signal nutritional status to the hypothalamus include the growth factors insulin and IGF-1. It is unclear which hypothalamic neuronal subpopulation these factors affect to ultimately regulate GnRH neuron function in puberty and reproduction. We examined the direct role of the GnRH neuron in growth factor regulation of reproduction using the Cre/lox system. Mice with the IR or IGF-1R deleted specifically in GnRH neurons were generated. Male and female mice with the IR deleted in GnRH neurons displayed normal pubertal timing and fertility, but male and female mice with the IGF-1R deleted in GnRH neurons experienced delayed pubertal development with normal fertility. With IGF-1 administration, puberty was advanced in control females, but not in females with the IGF-1R deleted in GnRH neurons, in control males, or in knockout males. These mice exhibited developmental differences in GnRH neuronal morphology but normal number and distribution of neurons. These studies define the role of IGF-1R signaling in the coordination of somatic development with reproductive maturation and provide insight into the mechanisms regulating pubertal timing in anabolic states.


Sodium ion binding pocket mutations and adenosine A2A receptor function.
Recently we identified a sodium ion binding pocket in a high-resolution structure of the human adenosine A2A receptor. In the present study we explored this binding site through site-directed mutagenesis and molecular dynamics simulations. Amino acids in the pocket were mutated to alanine, and their influence on agonist and antagonist affinity, allosterism by sodium ions and amilorides, and receptor functionality was explored. Mutation of the polar residues in the Na(+) pocket were shown to either abrogate (D52A(2.50) and N284A(7.49)) or reduce (S91A(3.39), W246A(6.48), and N280A(7.45)) the negative allosteric effect of sodium ions on agonist binding. Mutations D52A(2.50) and N284A(7.49) completely abolished receptor signaling, whereas mutations S91A(3.39) and N280A(7.45) elevated basal activity and mutations S91A(3.39), W246A(6.48), and N280A(7.45) decreased agonist-stimulated receptor signaling. In molecular dynamics simulations D52A(2.50) directly affected the mobility of sodium ions, which readily migrated to another pocket formed by Glu13(1.39) and His278(7.43). The D52A(2.50) mutation also decreased the potency of amiloride with respect to ligand displacement but did not change orthosteric ligand affinity. In contrast, W246A(6.48) increased some of the allosteric effects of sodium ions and amiloride, whereas orthosteric ligand binding was decreased. These new findings suggest that the sodium ion in the allosteric binding pocket not only impacts ligand affinity but also plays a vital role in receptor signaling. Because the sodium ion binding pocket is highly conserved in other class A G protein-coupled receptors, our findings may have a general relevance for these receptors and may guide the design of novel synthetic allosteric modulators or bitopic ligands.


Chronic pain patient and anaesthesia.
Severe chronic pain is often devastating for the affected individuals causing substantial suffering, health impairment, and a very low quality of life, including significant negative consequences for the patient and for society. Patients with complex pain disorders are seen often in relation to anaesthesia. They deserve special attention and require long time hospitalization and multiple contacts with health-care providers after discharge from hospital. A wider adoption of best perioperative and intraoperative pain management practice is required. This paper reviews current knowledge of perioperative and intraoperative pain management and anaesthetic care of the chronic pain patient. The individual topics covered include the magnitude of the problem created by chronic pain, the management of the patients taking various types of opioids, tolerance and opioid induced hyperalgesia and the multidisciplinary approach to pain management. The preventive and preemptive strategies for reducing the opioid needs and chronic pain after surgery are reviewed. The last section includes the role of acute pain services and an example of the implementation of a transitional pain service with the various benefits it offers; for example, the decrease of the opioid dose after discharge from the hospital. Patients also receive the continuity of care, with not only pain relief but also improvements in physical functioning, quality of life and emotional stress.


The reciprocal relationship between material factors and health in the life course: evidence from SHARE and ELSA.
The widely established health differences between people with greater economic resources and those with fewer resources can be attributed to both social causation (material factors affecting health) and health selection (health affecting material wealth). Each of these pathways may have different intensities at different ages, because the sensitivity of health to a lack of material wealth and the degree to which health can influence economic resources may change. We study the relative importance, in terms of explanatory power, of social causation and health selection, comparing the transitions from childhood to adulthood and from adulthood to old age. We use retrospective survey data from ten European countries from the Survey of Health, Ageing and Retirement in Europe (SHARELIFE, n = 18,734) and the English Longitudinal Study of Ageing (ELSA, n = 6117), and structural equations models in a cross-lagged panel design. Material wealth and health depend on their prior status, wealth more so than health. In the transition from childhood to adulthood, social causation and health selection are equally important: the standardized coefficients for men in SHARE are 0.07 and 0.06, respectively, i.e. one standard deviation increase in material wealth in childhood is associated with a 0.07 standard deviation increase in adult health. In the transition from adulthood to old age, social causation is more important than health selection (0.52 vs. 0.01), across gender and data sets. Both pathways contribute to the creation of health inequalities-however, their relative importance changes with age, which is important for understanding how health inequalities develop and how policies can address them.


Comparison of molecular dynamics and superfamily spaces of protein domain deformation.
It is well known the strong relationship between protein structure and flexibility, on one hand, and biological protein function, on the other hand. Technically, protein flexibility exploration is an essential task in many applications, such as protein structure prediction and modeling. In this contribution we have compared two different approaches to explore the flexibility space of protein domains: i) molecular dynamics (MD-space), and ii) the study of the structural changes within superfamily (SF-space). Our analysis indicates that the MD-space and the SF-space display a significant overlap, but are still different enough to be considered as complementary. The SF-space space is wider but less complex than the MD-space, irrespective of the number of members in the superfamily. Also, the SF-space does not sample all possibilities offered by the MD-space, but often introduces very large changes along just a few deformation modes, whose number tend to a plateau as the number of related folds in the superfamily increases. Theoretically, we obtained two conclusions. First, that function restricts the access to some flexibility patterns to evolution, as we observe that when a superfamily member changes to become another, the path does not completely overlap with the physical deformability. Second, that conformational changes from variation in a superfamily are larger and much simpler than those allowed by physical deformability. Methodologically, the conclusion is that both spaces studied are complementary, and have different size and complexity. We expect this fact to have application in fields as 3D-EM/X-ray hybrid models or ab initio protein folding.


Single d(GpG)/cis-diammineplatinum(II) adduct-induced inhibition of DNA polymerization.
A 44 nucleotide DNA template containing a single site-specifically placed cisplatin adduct (cis-[Pt(NH3)2[d(GpG)-N7(1),-N7(2)]]) was annealed with a primer, positioning its 3'-end four bases before the adduct in the template strand. DNA polymerization in the presence of all four nucleotides revealed that both HIV-1 reverse transcriptase (RT) and T7 DNA polymerase strongly paused at one nucleotide preceding the first platinated guanine and at the positions opposite the two platinated guanines. Analysis of single nucleotide incorporation at each pause site showed that polymerization occurs with biphasic kinetics. A small percentage of DNA was bound productively, providing a small amplitude (1-3%) of a fast phase of polymerization, whereas most of the bound DNA (1-34%) was positioned at the pause site in a nonproductive manner and therefore elongated slowly (0.04-0.06 s-1). DNA substrates annealed to the cisplatin-modified template bind to HIV-1 RT with an affinity (10-20 nM) similar to that of unmodified substrates (6-9 nM). The cisplatin-DNA cross-link moderately weakened DNA binding to T7 DNA polymerase (12-115 nM) but significantly slowed the rate of incorporation of the next nucleotide (2-7 s-1 ), with larger effects closer to the cisplatin-DNA adduct. The crystal structure of the same cisplatin-DNA adduct [Takahara, P. M., Frederick, C. A., and Lippard, S. J. (1996) J. Am. Chem. Soc. 118, 12309-12321] reveals not only the bent DNA duplex but also the propeller twisted base pairs near the cisplatin-DNA adduct. The twisted base pairs may cause misalignment of the cisplatin-modified DNA at the binding cleft of T7 DNA polymerase and significantly slow the rate of the protein conformational change preceding polymerization, leading to the slight accumulation of intermediates within five base pairs of the adduct. The ground-state binding of the next correct nucleotide to the enzyme.DNA complex was weakened by the adduct with T7 DNA polymerase but unchanged with HIV-1 RT at sites other than the three strong pause sites. Nucleotide binding to both enzymes at the three strong pause sites was significantly weaker and less selective.


Bis(N'-benzoyl-pyridine-4-carbohydrazide)(1,10-phenanthroline)copper(II) dinitrate.
In the title complex, [Cu(C(13)H(11)N(3)O(2))(2)(C(12)H(8)N(2))](NO(3))(2), the Cu(II )atom (site symmetry 2) is coordinated by four N atoms from one 1,10-phenanthroline and two hydrazine ligands, respectively. The hydrazine ligands coordinate to the Cu(II)atom by a pyridine N atom. These four atoms form a slightly distorted square-planar N(4) donor set. In the packing, two additional Cu⋯O inter-actions occur [Cu⋯O = 2.462 (2) Å], resulting in a typical Jahn-Teller-distorted octahedral environment around the Cu atom. N-H⋯O hydrogen bonds result in a three-dimensional network. The O atoms of the anion are disordered over two positions in a 0.68 (2):0.32 (2) ratio.


Subretinal Drusenoid Deposits in the Elderly in a Population-Based Study (the Montrachet Study).
The aim of this study was to investigate the prevalence of subretinal drusenoid deposits (SDD) and to identify associated factors in an elderly population. The participants of the population-based Montrachet study underwent an exhaustive ophthalmologic examination, including color fundus photography and macular spectral domain-optical coherence tomography (SD-OCT), coupled with infrared reflectance imaging. The presence of SDD and other age-related macular degeneration lesions, according to the European Eye Epidemiology SD-OCT classification of macular diseases, and subfoveal choroidal thickness were recorded. Moreover, the association of SDD and both clinical and demographic factors as well as plasma levels of vitamin E and lutein/zeaxanthin (L/Z) were analyzed. The mean age of patients was 82.3 ± 3.8 years and 62.7% were female. The prevalence of SDD was 18.1% (n = 205) in the subjects with at least one eye interpretable (n = 1135). In multivariate analysis, SDD was positively associated with increasing age (OR, 4.6; 95% CI, 2.8-7.7; P < 0.001 for subjects aged >85 years), female sex (OR, 1.7; 95% CI, 1.2-2.4; P = 0.005), and plasma L/Z level (OR, 1.2; 95% CI, 1.0-1.5; P = 0.039), and negatively associated with lipid-lowering drugs use (OR, 0.5; 95% CI, 0.3-0.9; P = 0.014 for statin medications) and subfoveal choroidal thickness (OR, 0.8; 95% CI, 0.7-0.9; P = 0.002). The prevalence of SDD was high in subjects older than 75 years, more frequent in women, and was associated with a thinner choroid. The association with lipid-lowering drugs deserves further investigation.


Volume of the amygdala is reduced in patients with narcolepsy - a structural MRI study.
Based on the clinical observation that patients suffering from narcolepsy with cataplexy (NC) have cataplectic attacks when they experience positive emotions, it is therefore hypothesised that the abnormal processing of external emotional input through the limbic system, or motor dysregulation induced by emotions, takes place during these episodes. To date, imaging studies have failed to reveal consistent brain abnormalities in NC patients. Considering the discrepancies in reported structural or functional abnormalities of the hypothalamus, amygdala, and nucleus accumbens, we used the MRI volumetry to determine the volumes of the amygdala and nucleus accumbens in a group of eleven patients with NC (5 males and 6 females, mean age 41.7 years ± 17.7). This data was compared to an equal number of examinations in healthy volunteers matched for age and gender. We found a decrease in the amygdalar volume of NC patients in both raw (p<0.001) and relative (p<0.01) data sets. The difference in amygdalar volume between healthy volunteers and NC patients was about 17%. In contrast to the amygdala, we did not find any differences in the volumes of nucleus accumbens. In the present MRI volumetric study, we found bilateral gray matter loss in the amygdala only.


Geometric Effect for Biological Reactors and Biological Fluids.
As expressed "God made the bulk; the surface was invented by the devil" by W. Pauli, the surface has remarkable properties because broken symmetry in surface alters the material properties. In biological systems, the smallest functional and structural unit, which has a functional bulk space enclosed by a thin interface, is a cell. Cells contain inner cytosolic soup in which genetic information stored in DNA can be expressed through transcription (TX) and translation (TL). The exploration of cell-sized confinement has been recently investigated by using micron-scale droplets and microfluidic devices. In the first part of this review article, we describe recent developments of cell-free bioreactors where bacterial TX-TL machinery and DNA are encapsulated in these cell-sized compartments. Since synthetic biology and microfluidics meet toward the bottom-up assembly of cell-free bioreactors, the interplay between cellular geometry and TX-TL advances better control of biological structure and dynamics in vitro system. Furthermore, biological systems that show self-organization in confined space are not limited to a single cell, but are also involved in the collective behavior of motile cells, named active matter. In the second part, we describe recent studies where collectively ordered patterns of active matter, from bacterial suspensions to active cytoskeleton, are self-organized. Since geometry and topology are vital concepts to understand the ordered phase of active matter, a microfluidic device with designed compartments allows one to explore geometric principles behind self-organization across the molecular scale to cellular scale. Finally, we discuss the future perspectives of a microfluidic approach to explore the further understanding of biological systems from geometric and topological aspects.


Targeting FK506 binding proteins to fight malarial and bacterial infections: current advances and future perspectives.
There is an urgent need for the design and development of new and selective drugs for the treatment of malaria and bacterial infections as these pathogens are developing resistance to presently available therapies. Malaria is a life threatening disease in many countries and responsible for almost one million deaths annually. In particular, drug-resistant malarial parasites are hindering effective control of malaria and prompting to find novel druggable targets and develop compounds with mechanism of action different from the conventional drugs. In this quest, efforts were made to determine three-dimensional structures of Plasmodium falciparum and Plasmodium vivax FK506 binding proteins which bind the macrolides (FK506 and rapamycin) and also demonstrate peptidylprolyl cis-trans isomerase activity in a similar manner as human FKBP12. Previous studies revealed that the immunosuppressive drug FK506 exhibits potential anti-malarial activity by binding FK506 binding domains (FKBD). This review focuses on three different types of FK506 binding proteins/domains in pathogens, their structural characteristics and biological roles. Binding ability of these proteins with the macrolides has opened new possibilities to develop selective inhibitors for these novel targets to combat the life threatening infections.


Myoglobin levels at 12 hours identify patients at low risk for 30-day mortality after thrombolysis in acute myocardial infarction: a Thrombolysis in Myocardial Infarction 10B substudy.
We sought to identify, by use of serum cardiac markers, patients at low risk for 30-day mortality after ST-segment elevation myocardial infarction. Baseline cardiac markers are currently used to identify patients at increased risk for short-term events. We hypothesized that serum markers measured after treatment could identify patients at low risk for 30-day mortality. A total of 839 patients from the Thrombolysis in Myocardial Infarction (TIMI) 10B study had myoglobin, cardiac-specific troponin-I, creatine kinase (CK)-MB measurements at the following time points; baseline, 90 minutes, and 3 and 12 hours after thrombolysis. By use of receiver operating characteristic analysis, thresholds were derived to predict 30-day mortality with at least 95% negative predictive value. Ninety minutes after thrombolysis myoglobin was superior to troponin-I or CK-MB in identifying patients at low risk for mortality. The 30-day mortality for 12-hour myoglobin < or = 239 ng/mL was 1.4% compared with 9.1% for levels > 239 ng/mL (P < .001). For 12-hour troponin-I (threshold 81.5 ng/mL), mortality was 1.9% versus 6.6% (P = .001) if above threshold; similarly for CK-MB at 12 hours (threshold 191 ng/mL) it was 3.3% versus 7.9% (P = .02). Multivariate analysis of baseline and posttreatment cardiac markers, age, sex, infarct artery location, and 90-minute TIMI flow grade identified only 12-hour myoglobin among the cardiac markers as independently predicting a low 30-day mortality (odds ratio 0.11, 95% confidence interval 0.02-0.50, P < .004). Serum cardiac markers can identify greater than two thirds of patients at low risk for 30-day mortality. A low 12-hour myoglobin level (< or = 239 ng/mL in this substudy) identifies such patients at low risk and could potentially assist in early risk stratification and triage after ST-segment elevation myocardial infarction.


A Systematic Review and Meta-analysis of Childhood Health Utilities.
A common feature of most reviews or catalogues of health utilities has been their focus on adult health states or derivation of values from adult populations. More generally, utility measurement in or on behalf of children has been constrained by several methodological concerns. The objective of this study was to conduct the first comprehensive systematic review and meta-analysis of primary utility data for childhood conditions and descriptors, and to determine the effects of methodological factors on childhood utilities. The review followed PRISMA guidelines. PubMed, Embase, Web of Science, PsycINFO, EconLit, CINAHL and Cochrane Library were searched for primary studies reporting health utilities for childhood conditions or descriptors using direct or indirect valuation methods. The Paediatric Economic Database Evaluation (PEDE) Porject was also searched for cost-utility analyses with primary utility values. Mean or median utilities for each of the main samples were catalogued, and weighted averages of utilities for each health condition were estimated, by valuation method. Mixed-effects meta-regression using hierarchical linear modeling was conducted for the most common valuation methods to estimate the utility decrement for each health condition category relative to general childhood population health, as well as the independent effects of methodological factors. The literature searches resulted in 272 eligible studies. These yielded 3,414 utilities when all sub-groups were considered, covering all ICD-10 chapters relevant to childhood health, 19 valuation methods, 12 respondent types, 8 modes of administration, and data from 36 countries. A total of 1,191 utility values were obtained when only main study samples were considered, and these were catalogued by health condition or descriptor, and methodological characteristics. 1,073 mean utilities for main samples were used for fixed-effects meta-analysis by health condition and valuation method. Mixed-effects meta-regressions estimated that 53 of 76 ICD-10 delineated health conditions, valued using the HUI3, were associated with statistically significant utility decrements relative to general population health, whereas 38 of 57 valued using a visual analog scale (VAS) were associated with statistically significant VAS decrements. For both methods, parental proxy assessment was associated with overestimation of values, whereas adolescents reported lower values than children under 12 y. VAS responses were more heavily influenced by mode of administration than the HUI3. Utilities and their associated distributions, as well as the independent contributions of methodological factors, revealed by this systematic review and meta-analysis can inform future economic evaluations within the childhood context.


Transmural flow modulates cell and fluid transport functions of lymphatic endothelium.
Lymphatic transport of peripheral interstitial fluid and dendritic cells (DCs) is important for both adaptive immunity and maintenance of tolerance to self-antigens. Lymphatic drainage can change rapidly and dramatically on tissue injury or inflammation, and therefore increased fluid flow may serve as an important early cue for inflammation; however, the effects of transmural flow on lymphatic function are unknown. Here we tested the hypothesis that lymph drainage regulates the fluid and cell transport functions of lymphatic endothelium. Using in vitro and in vivo models, we demonstrated that lymphatic endothelium is sensitive to low levels of transmural flow. Basal-to-luminal flow (0.1 and 1 mum/sec) increased lymphatic permeability, dextran transport, and aquaporin-2 expression, as well as DC transmigration into lymphatics. The latter was associated with increased lymphatic expression of the DC homing chemokine CCL21 and the adhesion molecules intercellular adhesion molecule-1 and E-selectin. In addition, transmural flow induced delocalization and downregulation of vascular endothelial cadherin and PECAM-1 (platelet/endothelial cell adhesion molecule-1). Flow-enhanced DC transmigration could be reversed by blocking CCR7, intercellular adhesion molecule-1, or E-selectin. In an experimental model of lymphedema, where lymphatic drainage is greatly reduced or absent, lymphatic endothelial expression of CCL21 was nearly absent. These findings introduce transmural flow as an important regulator of lymphatic endothelial function and suggest that flow might serve as an early inflammatory signal for lymphatics, causing them to regulate transport functions to facilitate the delivery of soluble antigens and DCs to lymph nodes.


E-learning in graduate medical education: survey of residency program directors.
E-learning-the use of Internet technologies to enhance knowledge and performance-has become a widely accepted instructional approach. Little is known about the current use of e-learning in postgraduate medical education. To determine utilization of e-learning by United States internal medicine residency programs, program director (PD) perceptions of e-learning, and associations between e-learning use and residency program characteristics. We conducted a national survey in collaboration with the Association of Program Directors in Internal Medicine of all United States internal medicine residency programs. Of the 368 PDs, 214 (58.2%) completed the e-learning survey. Use of synchronous e-learning at least sometimes, somewhat often, or very often was reported by 85 (39.7%); 153 programs (71.5%) use asynchronous e-learning at least sometimes, somewhat often, or very often. Most programs (168; 79%) do not have a budget to integrate e-learning. Mean (SD) scores for the PD perceptions of e-learning ranged from 3.01 (0.94) to 3.86 (0.72) on a 5-point scale. The odds of synchronous e-learning use were higher in programs with a budget for its implementation (odds ratio, 3.0 [95% CI, 1.04-8.7]; P = .04). Residency programs could be better resourced to integrate e-learning technologies. Asynchronous e-learning was used more than synchronous, which may be to accommodate busy resident schedules and duty-hour restrictions. PD perceptions of e-learning are relatively moderate and future research should determine whether PD reluctance to adopt e-learning is based on unawareness of the evidence, perceptions that e-learning is expensive, or judgments about value versus effectiveness.


A Subspace Pursuit-based Iterative Greedy Hierarchical solution to the neuromagnetic inverse problem.
Magnetoencephalography (MEG) is an important non-invasive method for studying activity within the human brain. Source localization methods can be used to estimate spatiotemporal activity from MEG measurements with high temporal resolution, but the spatial resolution of these estimates is poor due to the ill-posed nature of the MEG inverse problem. Recent developments in source localization methodology have emphasized temporal as well as spatial constraints to improve source localization accuracy, but these methods can be computationally intense. Solutions emphasizing spatial sparsity hold tremendous promise, since the underlying neurophysiological processes generating MEG signals are often sparse in nature, whether in the form of focal sources, or distributed sources representing large-scale functional networks. Recent developments in the theory of compressed sensing (CS) provide a rigorous framework to estimate signals with sparse structure. In particular, a class of CS algorithms referred to as greedy pursuit algorithms can provide both high recovery accuracy and low computational complexity. Greedy pursuit algorithms are difficult to apply directly to the MEG inverse problem because of the high-dimensional structure of the MEG source space and the high spatial correlation in MEG measurements. In this paper, we develop a novel greedy pursuit algorithm for sparse MEG source localization that overcomes these fundamental problems. This algorithm, which we refer to as the Subspace Pursuit-based Iterative Greedy Hierarchical (SPIGH) inverse solution, exhibits very low computational complexity while achieving very high localization accuracy. We evaluate the performance of the proposed algorithm using comprehensive simulations, as well as the analysis of human MEG data during spontaneous brain activity and somatosensory stimuli. These studies reveal substantial performance gains provided by the SPIGH algorithm in terms of computational complexity, localization accuracy, and robustness.


Novel modulating effects of PKC family genes on the relationship between serum vitamin D and relapse in multiple sclerosis.
The interplay between genes and environmental factors on multiple sclerosis (MS) clinical course has been little studied. We conducted a prospective cohort study of 141 participants with relapsing-remitting MS (RRMS) and genotype data followed from 2002 to 2005 and examined genes in the vitamin D metabolism and vitamin D receptor (VDR)/retinoid X receptor (RXR) transcription factor formation pathway. Gene-vitamin D interactions and the genetic predictors of relapse were assessed using survival analysis. Genetic predictors of 25-hydroxyvitamin D (25(OH)D) were evaluated by multilevel mixed-effects linear regression. Significance threshold was adjusted by Bonferroni correction for the number of genes evaluated. The relationship between 25(OH)D and hazard of relapse was significantly different for different alleles of two intronic single nucleotide polymorphisms (SNPs) (rs908742 in PRKCZ and rs3783785 in PRKCH) in the protein kinase C (PKC) family genes (p(interaction)=0.001, p(adj)=0.021, respectively). Two other intronic SNPs (rs1993116 in CYP2R1and rs7404928 in PRKCB) were significantly associated with lower levels of 25(OH)D (p(interaction)=0.001, p(adj)=0.021, respectively). A cumulative effect of multiple 'risk' genotypes on 25(OH)D levels and hazard of relapse was observed for the significant SNPs (p(trend)=7.12×10(-6) for 25(OH)D levels, p(trend)=8.86×10(-6) for hazard of relapse). Our data support the hypothesis that gene-vitamin D interactions may influence MS clinical course and that the PKC family genes may play a role in the pathogenesis of MS relapse through modulating the association between 25(OH)D and relapse.


Direct quantitative transcript analysis of the agr regulon of Staphylococcus aureus during human infection in comparison to the expression profile in vitro.
Bacteria possess a repertoire of distinct regulatory systems promoting survival in disparate environments. Under in vitro conditions it was demonstrated for the human pathogen Staphylococcus aureus that the expression of most virulence factors is coordinated by the global regulator agr. To monitor bacterial gene regulation in the host, we developed a method for direct transcript analysis from clinical specimens. Quantification of specific transcripts was performed by competitive reverse transcription-PCR, and results were normalized against the constitutively expressed gene for gyrase (gyr). Using sputum from cystic fibrosis (CF) patients infected with S. aureus we examined the transcription of the effector molecule RNAIII of agr, of spa (protein A), generally repressed by agr, and of hla (alpha-toxin), generally activated by agr. In the CF lung RNAIII was expressed poorly, indicating an inactive agr in vivo. Despite the low level of RNAIII expression, spa was detectable only in minute amounts and an irregular transcription of hla was observed in all sputum samples. After subculturing of patient strains agr-deficient isolates and isolates with unusual expression profiles, i.e., not consistent with those obtained from prototypic strains, were observed. In conclusion, the agr activity seems to be nonessential in CF, and from the described expression pattern of spa and hla, other regulatory circuits aside from agr are postulated in vivo.


Insulin-transferrin-selenium prevent human chondrocyte dedifferentiation and promote the formation of high quality tissue engineered human hyaline cartilage.
This study was to investigate the effects of insulin-transferrin-selenium (ITS) on the proliferation and quantitative gene expression of adult human nasal septum chondrocytes in monolayer culture expansion and the formation of tissue engineered hyaline cartilage. Effects of ITS on human nasal septum chondrocytes monolayer culture expansion and gene expression were evaluated in various culture media either added with 2% fetal bovine serum (FBS) or 1 ng/mL basic fibroblast growth factor plus 1 ng/mL transforming growth factor or both serum and growth factors supplementation in comparison with medium added with 10%FBS. Chondrocytes cultured in medium added with 2% fetal bovine serum and growth factors either supplemented with or without ITS were then mixed with pluronic F-127 hydrogel for in vivo tissue engineered cartilage formation in nude mice model. Engineered tissues were removed after 8 weeks of implantation and evaluated with histological staining, immunohistochemistry, transmission electron microscopy and quantitative gene expression analysis. ITS promoted human chondrocytes proliferation and reduced chondrocytes dedifferentiation in media supplemented with serum and growth factors. ITS with 2% FBS and growth factors provided 15-fold increased in chondrocytes number by the end of the culture period compared to the standard culture medium used in chondrocytes culture (medium added with 10% FBS). Engineered tissue resulted from ITS supplementation demonstrated higher quality of cartilage formation. In conclusion, our study has demonstrated the benefits of ITS supplementation in human chondrocytes monolayer culture and tissue engineering cartilage formation.


Kinetic silver staining and quantification of proteins adsorbed to microtiter plates.
A silver stain was used to detect and quantitate proteins adsorbed to microtiter plate wells. The kinetics of the development of the silver stain were analyzed with an automated microtiter plate reader. The lag time for stain development was found to be a consistent indicator of the amount of protein adsorbed to a microtiter plate well. Protein which was not preadsorbed to the microtiter plate was not effectively stained by silver. Complete adsorption of protein applied to the microtiter plate was possible by drying small amounts of protein in very dilute buffers. Variations in sensitivity for different proteins were less than 30% for the panel of proteins examined. Determinations from kinetic silver staining agreed with those from copper staining for bovine albumin adsorbed to microtiter plates. The precision of kinetic silver staining assay was optimal in the range of 40 to 200 ng per microtiter plate well. In this range, the standard deviations averaged less than 5%. Even smaller amounts of protein can be detected and interpolated down to approximately 10 ng per well. The kinetic silver staining method can be used on standard microtiter plate readers without special filters and is readily adaptable to automated systems.


Acetylcholinesterase/paraoxonase genotype and expression predict anxiety scores in Health, Risk Factors, Exercise Training, and Genetics study.
Anxiety involves complex, incompletely understood interactions of genomic, environmental, and experience-derived factors, and is currently being measured by psychological criteria. Here, we report previously nonperceived interrelationships between expression variations and nucleotide polymorphisms of the chromosome 7q21-22 acetylcholinesterase-paraoxonase 1 (ACHE-PON1) locus with the trait- and state-anxiety measures of 461 healthy subjects from the Health, Risk Factors, Exercise Training, and Genetics Family Study. The AChE protein controls the termination of the stress-enhanced acetylcholine signaling, whereas the PON protein displays peroxidase-like activity, thus protecting blood proteins from oxidative stress damages. Serum AChE and PON enzyme activities were both found to be affected by demographic parameters, and showed inverse, reciprocal associations with anxiety measures. Moreover, the transient scores of state anxiety and the susceptibility score of trait anxiety both appeared to be linked to enzyme activities. This finding supported the notion of corresponding gene expression relationships. Parallel polymorphisms in the ACHE and PON1 genes displayed apparent associations with both trait- and state-anxiety scores. Our findings indicate that a significant source of anxiety feelings involves inherited and acquired parameters of acetylcholine regulation that can be readily quantified, which can help explaining part of the human variance for state and trait anxiety.


Volumetric finite-element modelling of biological growth.
Differential growth is the driver of tissue morphogenesis in plants, and also plays a fundamental role in animal development. Although the contributions of growth to shape change have been captured through modelling tissue sheets or isotropic volumes, a framework for modelling both isotropic and anisotropic volumetric growth in three dimensions over large changes in size and shape has been lacking. Here, we describe an approach based on finite-element modelling of continuous volumetric structures, and apply it to a range of forms and growth patterns, providing mathematical validation for examples that admit analytic solution. We show that a major difference between sheet and bulk tissues is that the growth of bulk tissue is more constrained, reducing the possibility of tissue conflict resolution through deformations such as buckling. Tissue sheets or cylinders may be generated from bulk shapes through anisotropic specified growth, oriented by a polarity field. A second polarity field, orthogonal to the first, allows sheets with varying lengths and widths to be generated, as illustrated by the wide range of leaf shapes observed in nature. The framework we describe thus provides a key tool for developing hypotheses for plant morphogenesis and is also applicable to other tissues that deform through differential growth or contraction.


A review of e-textiles in neurological rehabilitation: How close are we?
Textiles able to perform electronic functions are known as e-textiles, and are poised to revolutionise the manner in which rehabilitation and assistive technology is provided. With numerous reports in mainstream media of the possibilities and promise of e-textiles it is timely to review research work in this area related to neurological rehabilitation.This paper provides a review based on a systematic search conducted using EBSCO- Health, Scopus, AMED, PEDro and ProQuest databases, complemented by articles sourced from reference lists. Articles were included if the e-textile technology described had the potential for use in neurological rehabilitation and had been trialled on human participants. A total of 108 records were identified and screened, with 20 meeting the broad review inclusion criteria. Nineteen user trials of healthy people and one pilot study with stroke participants have been reported.The review identifies two areas of research focus; motion sensing, and the measurement of, or stimulation of, muscle activity. In terms of motion sensing, E-textiles appear able to reliably measure gross movement and whether an individual has achieved a predetermined movement pattern. However, the technology still remains somewhat cumbersome and lacking in resolution at present. The measurement of muscle activity and the provision of functional electrical stimulation via e-textiles is in the initial stages of development but shows potential for e-textile expansion into assistive technologies.The review identified a lack of high quality clinical evidence and, in some cases, a lack of practicality for clinical application. These issues may be overcome by engagement of clinicians in e-textile research and using their expertise to develop products that augment and enhance neurological rehabilitation practice.


Interpreting the value of feedback: Older adult voices in nursing education.
As patients, older adults are often involved informally with the teaching of nursing students in clinical environments. This involvement is recognised as valuable; however, the role of the patient is often passive. In recent years there has been recognition of the value that real people's lives, and health experiences can bring to student education, so much so that many programs now incorporate real patients into the formal education of students. In this educational initiative, older adults, from a Residential Aged Care Campus (RACC), participated as simulated patients in scenario based clinical learning activities, providing feedback to nursing students on the carative aspects of the learning activities. The aim of this study was to contribute toward the body of knowledge in engaging older adults in simulated clinical activities, specifically in the provision of feedback to nursing students. A qualitative interpretive methodology was applied to capture the older adults' experiences of providing feedback as simulated patients. This included the collection of data through participant observation, recording of field notes, collection of a written feedback form and semi-structured interviews with the participants. Thematic analysis was undertaken to identify emergent themes in the data. Findings from this study have emphasised the value of including the patient voice in nursing education. The provision of feedback by older adults during direct care interactions supports and nurtures student awareness of the patient and the caring process of nursing. Unique to this study, was the finding that although the participants received no formal training in the delivery of feedback, all utilised a clear process recognised within the literature. A recommendation linked to this finding is that a feedback training program would be of benefit in reducing some of the feelings of uncertainty expressed by the older adults when giving feedback and better equipping them to deliver constructive feedback.


Responses of 6500 households to arsenic mitigation in Araihazar, Bangladesh.
This study documents the response of 6500 rural households in a 25 km(2) area of Bangladesh to interventions intended to reduce their exposure to arsenic contained in well water. The interventions included public education, posting test results for arsenic on the wells, and installing 50 community wells. Sixty-five percent of respondents from the subset of 3410 unsafe wells changed their source of drinking water, often to new and untested wells. Only 15% of respondents from the subset of safe wells changed their source, indicating that health concerns motivated the changes. The geo-referenced data indicate that distance to the nearest safe well also influenced household responses.


Multidimensional Processes: In Italy, biomedical signal and image processing embraces a multiparametric, multimodal, multiscale paradigm.
Biomedical studies, both in research and in clinical applications, deal with the management of large amounts of data. Different sensors and transducers, advances in technologies, and the availability of innovative medical equipment and instrumentation all contribute to the ability to make biological measurements at different scales, ranging from systems, to organs, to tissues, to cells, right down to proteins and genes. Biomedical signals and data carry important information about the system or the organ that generated them.


Transcranial Direct Current Stimulation Accelerates The Onset of Exercise-Induced Hypoalgesia: A Randomized Controlled Study.
Exercise-induced hypoalgesia (EIH) describes acute reductions in pain that occur following exercise. Current evidence suggests that the magnitude of EIH is small-to-moderate at best, warranting exploration of novel avenues to bolster these effects. Transcranial direct current stimulation (tDCS) has been shown to relieve pain and represents a promising intervention that may enhance EIH. This study aimed to determine whether anodal tDCS of the primary motor cortex (M1) can augment EIH in healthy individuals experiencing experimentally-induced musculoskeletal pain. Twenty-four healthy subjects attended 2 experimental sessions ("Day 0" and "Day 2"). On Day 0, subjects were injected with nerve growth factor into their right extensor carpi radialis brevis to induce persistent elbow pain. On Day 2, each subject received active or sham tDCS over M1 followed by an isometric grip exercise. Pain intensity, muscle soreness, sensitivity (pressure pain thresholds), and conditioned pain modulation were assessed prior to the nerve growth factor injection, on Day 2 before tDCS, immediately post-exercise, and 15 minutes post-exercise. Active tDCS expedited the onset of EIH, inducing immediate reductions in pain intensity that were not present until 15 minutes post-exercise in the sham group. However, active tDCS did not reduce muscle soreness or sensitivity when compared to sham tDCS. PERSPECTIVE: These findings suggest that active tDCS accelerates the onset of EIH in healthy individuals experiencing experimentally-induced pain. This may represent a promising means of enhancing adherence to exercise protocols. However, larger randomised controlled trials in persistent pain populations are required to confirm the clinical impact of these findings.


The effects of the visual environment on responses to colour by domestic chicks.
It is well known that development of vision is affected by experience, but there are few studies of environmental effects on colour vision. Natural scenes contain predominantly a restricted range of reflectance spectra, so such effects might be important, perhaps biasing visual mechanisms towards common colours. We investigated how the visual environment affects colour preferences of domestic chicks ( Gallus gallus), by training week-old birds to select small food containers distinguished from an achromatic alternative either by an orange or by a greenish-blue colour. Chicks that had been raised in control conditions, with long-wavelength-dominated reflectance spectra, responded more readily to orange than to blue. This was not due to avoidance of blue, as increasing saturation enhanced the chicks' preference for the same hue. The advantage of orange was, however, reduced or abolished for chicks raised in an environment dominated by blue objects. This indicates that responses to coloured food are affected by experience of non-food objects. If colours of ordinary objects in the environment do influence responses to specialised visual signals this might help explain why biological signals directed at birds are often coloured yellow, orange or red; long-wavelength-dominated spectra being more prevalent than short-wavelength-dominated spectra.


Alveolar type II cells escape stress failure caused by tonic stretch through transient focal adhesion disassembly.
Mechanical ventilation-induced excessive stretch of alveoli is reported to induce cellular stress failure and subsequent lung injury, and is therefore an injurious factor to the lung. Avoiding cellular stress failure is crucial to ventilator-induced lung injury (VILI) treatment. In the present study, primary rat alveolar type II (ATII) cells were isolated to evaluate their viability and the mechanism of their survival under tonic stretch. By the annexin V/ PI staining and flow cytometry assay, we demonstrated that tonic stretch-induced cell death is an immediate injury of mechanical stress. In addition, immunofluorescence and immunoblots assay showed that the cells experienced an expansion-contraction-reexpansion process, accompanied by partial focal adhesion (FA) disassembly during contraction. Manipulation of integrin adherent affinity by altering bivalent cation levels in the culture medium and applying an integrin neutralizing antibody showed that facilitated adhesion affinity promoted cell death under tonic stretch, while lower level of adhesion protected the cells from stretch-induced stress failure. Finally, a simplified numerical model was established to reveal that adequate disassembly of FAs reduced the forces transmitting throughout the cell. Taken together, these results indicate that ATII cells escape stress failure caused by tonic stretch via active cell morphological remodeling, during which cells transiently disassemble FAs to unload mechanical forces.


Effects of mutations near the bacteriochlorophylls in reaction centers from Rhodobacter sphaeroides.
Mutations were made in four residues near the bacteriochlorophyll cofactors of the photosynthetic reaction center from Rhodobacter sphaeroides. These mutations, L131 Leu to His and M160 Leu to His, near the dimer bacteriochlorophylls, and M203 Gly to Asp and L177 Ile to Asp, near the monomer bacteriochlorophylls, were designed to result in the placement of a hydrogen bond donor group near the ring V keto carbonyl of each bacteriochlorophyll. Perturbations of the electronic structures of the bacteriochlorophylls in the mutants are indicated by additional resolved transitions in the bacteriochlorophyll absorption bands in steady-state low-temperature and time-resolved room temperature spectra in three of the resulting mutant reaction centers. The major effect of the two mutations near the dimer was an increase up to 80 mV in the donor oxidation-reduction midpoint potential. Correspondingly, the calculated free energy difference between the excited state of the primary donor and the initial charge separated state decreased by up to 55 mV, the initial forward electron-transfer rate was up to 4 times slower, and the rate of charge recombination between the primary quinone and the donor was approximately 30% faster in these two mutants compared to the wild type. The two mutations near the monomer bacteriochlorophylls had minor changes of 25 mV or less in the donor oxidation-reduction potential, but the mutation close to the monomer bacteriochlorophyll on the active branch resulted in a roughly 3-fold decrease in the rate of the initial electron transfer.


Using absolute and relative muscle endurance to estimate maximal strength in young athletes.
The purpose of this study was to evaluate the effectiveness of repetitions to fatigue (RTF) using absolute and relative muscle-endurance performances to estimate 1-repetition-maximum (1-RM) bench-press performance in high school male athletes. Members of high school athletic teams (n = 118, age = 16.5 +/- 1.1 y, weight = 82.7 +/- 18.7 kg) were tested for 1-RM bench press and RTF with an absolute load of 61.4 kg and a relative load that produced 7 to 10 RTF (7- to 10-RM). All participants had completed a minimum of 4 wk of resistance training before measurement. All 7- to 10-RM-prediction equations had higher correlations between predicted and actual 1-RM (r > .98) than the 61.4-kg absolute-load equation (r = .95). Despite the high correlations, only 3 of 11 equations produced predicted values that were nonsignificantly different from actual 1-RM. The best 7-to 10-RM equation predicted 65% of the athletes'performances within +/-4.5 kg of their actual 1-RM. The addition of simple anthropometric dimensions did not increase the validity correlations or decrease the prediction errors. The 7- to 10-RM method can provide an accurate method of estimating strength levels for adjusting loads in a training program and is more accurate for predicting 1-RM bench press in high school athletes than the 61.4-kg repetition method.


Dynamic integration of conceptual information during learning.
The development and application of concepts is a critical component of cognition. Although concepts can be formed on the basis of simple perceptual or semantic features, conceptual representations can also capitalize on similarities across feature relationships. By representing these types of higher-order relationships, concepts can simplify the learning problem and facilitate decisions. Despite this, little is known about the neural mechanisms that support the construction and deployment of these kinds of higher-order concepts during learning. To address this question, we combined a carefully designed associative learning task with computational model-based functional magnetic resonance imaging (fMRI). Participants were scanned as they learned and made decisions about sixteen pairs of cues and associated outcomes. Associations were structured such that individual cues shared feature relationships, operationalized as shared patterns of cue pair-outcome associations. In order to capture the large number of possible conceptual representational structures that participants might employ and to evaluate how conceptual representations are used during learning, we leveraged a well-specified Bayesian computational model of category learning [1]. Behavioral and model-based results revealed that participants who displayed a tendency to link experiences in memory benefitted from faster learning rates, suggesting that the use of the conceptual structure in the task facilitated decisions about cue pair-outcome associations. Model-based fMRI analyses revealed that trial-by-trial integration of cue information into higher-order conceptual representations was supported by an anterior temporal (AT) network of regions previously implicated in representing complex conjunctions of features and meaning-based information.


Orthotomic systems of rays in inhomogeneous isotropic media.
The behavior of orthotomic systems of rays, rays associated with a system of wavefronts, is analyzed from the point of view of classical geometrical optics. The rays themselves are described in terms of the ray equation derived from Fermat's principle. A condition for an aggregate of rays to comprise an orthotomic system is found. Some consequences of this condition on the geometric properties of wave-fronts are found. The resemblance of some of these to the Maxwell equations is noted.


Do women fare worse? A metaanalysis of gender differences in outcome after traumatic brain injury.
The purpose of this metaanalysis was to investigate possible gender differences in TBI sequelae. The case fatality rates in patients after TBI have previously been shown to be significantly higher in women as compared with men. A quantitative review of published studies of TBI outcome revealed eight studies (20 outcome variables) of TBI in which outcome was reported separately for men and women. Outcome was worse in women than in men for 85% of the measured variables, with an average effect size of -0.15. Although clinical opinion is often that women tend to experience better outcomes than do men after TBI, the opposite pattern was suggested in the results of this metaanalysis. However, this conclusion is limited by the fact that in only a small percentage of the total published reports on TBI outcome was outcome described separately for each sex. A careful, prospective study of sex differences in TBI outcome is clearly needed.


The TLR4 D299G and T399I SNPs are constitutively active to up-regulate expression of Trif-dependent genes.
Dysregulated Toll-Like Receptor (TLR) signalling and genetic polymorphisms in these proteins are linked to many human diseases. We investigated TLR4 functional variants D299G and T399I to assess the impact on LPS-induced responsiveness in comparison to wild-type TLR4. The mechanism by which this occurs in unclear as these SNPs do not lie within the lipid A binding domain or dimerisation sites of the LPS-TLR4/MD2 receptor complexes. Transfection of TLR4D299G, TLR4T399I or TLR4D299G. T399I into HEK cells resulted in constitutive activation of an NF-κB reporter gene and a blunting of the LPS-induced reporter activation compared to WT-TLR4. Unstimulated human monocyte/macrophages, from patients with the D299G and T399I SNPs demonstrated a downregulation of many genes, particularly Tram/Trif signalling pathway constitutents compared to the TLR4 wild-type subjects supporting the concept of basal receptor activity. Monocyte/macrophages from carriers of the TLR4 D299G and T399I polymorphisms stimulated with LPS showed >6 fold lower levels of NF-κB and ∼12 fold higher IFN-β gene expression levels compared to wild-type subjects (P<0.05; MWU test) and dramatically altered resultant cytokine profiles. We conclude that these TLR4 SNPs affect constitutive receptor activity which impacts on the hosts ability to respond to LPS challenge leading to a dysregulated sub-optimal immune response to infection.


Weight gain is associated with reduced striatal response to palatable food.
Consistent with the theory that individuals with hypofunctioning reward circuitry overeat to compensate for a reward deficit, obese versus lean humans have fewer striatal D2 receptors and show less striatal response to palatable food intake. Low striatal response to food intake predicts future weight gain in those at genetic risk for reduced signaling of dopamine-based reward circuitry. Yet animal studies indicate that intake of palatable food results in downregulation of D2 receptors, reduced D2 sensitivity, and decreased reward sensitivity, implying that overeating may contribute to reduced striatal responsivity. Thus, we tested whether overeating leads to reduced striatal responsivity to palatable food intake in humans using repeated-measures functional magnetic resonance imaging. Results indicated that women who gained weight over a 6 month period showed a reduction in striatal response to palatable food consumption relative to weight-stable women. Collectively, results suggest that low sensitivity of reward circuitry increases risk for overeating and that this overeating may further attenuate responsivity of reward circuitry in a feedforward process.


Review article: S-adenosyl-L-methionine--a new therapeutic agent in liver disease?
The established biochemical effects of exogenous S-Adenosyl-L-Methionine (SAMe) are diverse and are still being explored in liver disease. Putative therapeutic effects could be exerted via different mechanisms. The established deficiency of SAMe synthetase in cirrhosis could by bypassed by exogenous SAMe, leading to increased levels of sulphur-containing amino acids and glutathione which would protect against oxidant stress and drug-induced hepatotoxicity (for example, paracetamol). Furthermore SAMe could act by improving membrane fluidity, and thus potentially improve or restore the function of receptors, enzymes and transporters in the cell surface. Membrane fluidity is known to be affected by alterations in cell membrane lipid composition in chronic liver disease. Very few therapeutic agents are effective for the symptomatic or specific treatment of chronic liver disease. SAMe has established biochemical and biophysical effects which in pilot studies ameliorate symptoms and biochemical parameters of cholestasis. Moreover, abnormalities in liver function tests (including transaminase values) also improve. Before SAMe can be considered as an established therapy for patients with hepatic disease, long-term controlled clinical trials of SAMe are needed to assess the benefit for patients' symptoms, well being, histological changes and progression of liver disease.


Transformation of 17alpha-ethinylestradiol during water chlorination: effects of bromide on kinetics, products, and transformation pathways.
Kinetics, products, and pathways of 17alpha-ethinylestradiol (EE2) transformation during chlorination of water with and without bromide (Br(-)) were studied to evaluate the effect of chlorination on the fate of steroid estrogens. Br(-) can be important for the fate of EE2 during chlorination because of a rapid conversion of Br(-) to bromine by chlorine and a approximately 1000 times higher reactivity of bromine than chlorine toward EE2. Experimental data and model calculations for EE2 chlorination show that 4-chloro EE2 and 2,4-dichloro EE2 are the major initial transformation products in absence of Br(-). With increasing concentration of Br(-), the bromo-analogous of the chloro EE2s becomes significant as additional initial transformation products. Those initial transformation products of EE2 are generally quickly further transformed to products with a destroyed phenolic moiety. Transformation of EE2 during chlorination becomes considerably faster at Br(-) concentrations higher than 0.25 microM (20 microg L(-1)). However, in presence of model dissolved organic matter such as glycine or phenol, the accelerating effect of Br(-) diminishes due to a more rapid consumption of bromine than chlorine by these compounds. A kinetic model was developed which allows to predict EE2 transformation during chlorination of natural waters and wastewaters in presence of Br(-) and low concentrations of ammonia (<10 microM). It could be demonstrated in these systems that bromine produced from Br(-) is mainly responsible for EE2 transformation.


Use of the probability of stone formation (PSF) score to assess stone forming risk and treatment response in a cohort of Brazilian stone formers.
The aim was to confirm that PSF (probability of stone formation) changed appropriately following medical therapy on recurrent stone formers. Data were collected on 26 Brazilian stone-formers. A baseline 24-hour urine collection was performed prior to treatment. Details of the medical treatment initiated for stone-disease were recorded. A PSF calculation was performed on the 24 hour urine sample using the 7 urinary parameters required: voided volume, oxalate, calcium, urate, pH, citrate and magnesium. A repeat 24-hour urine sample was performed for PSF calculation after treatment. Comparison was made between the PSF scores before and during treatment. At baseline, 20 of the 26 patients (77%) had a high PSF score (> 0.5). Of the 26 patients, 17 (65%) showed an overall reduction in their PSF profiles with a medical treatment regimen. Eleven patients (42%) changed from a high risk (PSF > 0.5) to a low risk (PSF < 0.5) and 6 patients reduced their risk score but did not change risk category. Six (23%) patients remained in a high risk category (> 0.5) during both assessments. The PSF score reduced following medical treatment in the majority of patients in this cohort.


Exercise- and allergen-induced asthma do not change the production of Paf-acether by neutrophils and platelets.
Paf-acether, whose role has been suggested in asthma, is a mediator released by stimulated neutrophils, platelets and other cells. Neutrophils and platelets are activated in vivo during exercise or allergen-induced asthma. Upon in vitro stimulation, macrophages from mice treated with an inflammatory stimulus, such as thioglycoccollate, release less paf-acether than macrophages from non-treated mice. We hypothesized that upon in vitro activation platelets and neutrophils should produce less paf-acether after exercise- or allergen-induced asthma. To test this hypothesis, we measured the production of paf-acether by neutrophils and platelets obtained before, 15 and 75 min after exercise in seven normal subjects and five asthmatic subjects with exercise-induced asthma, and in five other asthmatic subjects after specific challenge with Dermatophagoides Pteronyssinus. Purified neutrophils and washed platelets were incubated independently for 10 min at 37 degrees C with no specific activator, with a platelet activator (thrombin, 1 IU.ml-1), a neutrophil activator (opsonized zymosan, 1 mg.ml-1), and both together. We found no significant difference between asthmatic and normal subjects in the amount of paf-acether synthesized by platelets or neutrophils and no fall in the production of paf-acether after exercise- or allergen-induced asthma. However, our method may lack sensitivity in detecting partial activation of these cells and is based on the assumption that changes in peripheral blood cells are representative of changes of these cells in lungs.


Coactivator peptides have a differential stabilizing effect on the binding of estrogens and antiestrogens with the estrogen receptor.
The effectiveness of estrogens in stimulating gene transcription mediated by the estrogen receptor (ER) appears to depend on ER interactions with coactivator proteins. These coactivators bind to ER when it is liganded with an estrogen agonist, but not when it is liganded with an estrogen antagonist. Because estrogen agonists are known to induce a conformation in ER that stabilizes coactivator binding, we asked whether coactivator binding to ER causes a reciprocal stabilization of agonist ligand binding. We used a fluorescent ligand for ER, tetrahydrochrysene-ketone, to monitor the rates of ligand dissociation from ERalpha and ERbeta, and to see how this process is affected by the p160-class coactivator, steroid receptor coactivator-1 (SRC-1). We used a 15-amino acid peptide corresponding to the second nuclear receptor box LXXLL motif in SRC-1 (NR-2 peptide), which is known to interact with the ER ligand-binding domain, a mutant peptide with an LXXAL sequence (NR-2A peptide), and a 203-amino acid fragment of SRC-1, termed the nuclear receptor domain (SRC1-NRD), embodying all three of the internal NR boxes of this protein. Both the NR-2 peptide and the SRC1-NRD fragment markedly slow the rate of dissociation of the agonist ligands tetrahydrochrysene-ketone, estradiol, and diethylstilbestrol, increasing the half-life of the ER-agonist complex by up to 50- to 60-fold. The SRC1-NRD has much higher potency in retarding ligand dissociation than does the NR-2 peptide; it is maximally effective at 30 nM, and it appears to bind with the stoichiometry of one SRC1-NRD per ER dimer. The peptides had little effect on the dissociation rate of antagonist ligands. Consistent with these results, we find that increasing the concentration of SRC-1 in cells by transfection of an expression plasmid encoding SRC-1 causes a 17-fold increase in the potency of estradiol in an estrogen-responsive reporter gene transcription assay. Thus, there is multifactorial control over receptor-coactivator interaction, its strength being determined by the agonist vs. antagonist nature of the ligand and the particular structure of the agonist ligand, and by the receptor subtype and the NR box sequence. The stabilizing effect of coactivator on ER-agonist ligand complexes may be important in determining the potency of estrogen agonists in a cell and may also underlie the tissue-selective pharmacology of certain synthetic estrogens.


Seismic vulnerability assessment to earthquake at urban scale: A case of Mostaganem city in Algeria.
The focus of this study was the seismic vulnerability assessment of buildings constituting Mostaganem city in Algeria. Situated 320 km to the west of Algiers, Mostaganem city encompasses a valuable cultural and architectural built heritage. The city has suffered several moderate earthquakes in recent years; this has led to extensive structural damage to old structures, especially unreinforced historical buildings. This study was divided into two essential steps, the first step being to establish fragility curves based on a non-linear static pushover analysis for each typology and height of buildings. Twenty-seven pushover analyses were performed by means of SAP2000 software (three analyses for each type of building). The second step was to adopt the US HAZUS software and to modify it to suit the typical setting and parameters of the city of Mostaganem. A seismic vulnerability analysis of Mostaganem city was conducted using HAZUS software after inputting the new parameters of the fragility curves established within the first step. The results indicated that the number of poor-quality buildings expected to be totally destroyed under a 5.5 Mw earthquake scenario could reach more than 28 buildings. Three percent of unreinforced masonry (URM) buildings were completely damaged and 10% were extensively damaged. Of the concrete frame buildings, 6% were extensively damaged and 19% were moderately damaged. According to the built year, 6% of both concrete frame and URM buildings built before 1980 are estimated to be collapsing. Buildings constructed between 1980 and 1999 are more resistant; 8% of those structures were extensively damaged and 18% were moderately damaged. Only 10% of buildings constructed after 1999 were moderately damaged. The results also show that the main hospital of the city, built before 1960, will be extensively damaged during an earthquake of 5.5 Mw. The number of human casualties could reach several hundreds - 10.5% of residents of URM buildings are injured or dead. Compared with the URM buildings, concrete frame buildings have lower casualty rates of 1.5% and 0.5% for those built before and after 1980, respectively. It was concluded that Mostaganem city belongs to seismic vulnerable zones in Algeria; in this regard, an action plan is needed for the rehabilitation of old constructions. In addition, the effectiveness of establishing and introducing new and appropriate fragility curves was demonstrated.


Association of socioeconomic deprivation with life expectancy and all-cause mortality in Spain, 2011-2013.
Life tables summarise a population's mortality experience during a time period. Sex- and age-specific life tables are needed to compute various cancer survival measures. However, mortality rates vary according to socioeconomic status. We present sex- and age-specific life tables based on socioeconomic status at the census tract level in Spain during 2011-2013 that will allow estimating cancer relative survival estimates and life expectancy measures by socioeconomic status. Population and mortality data were obtained from the Spanish Statistical Office. Socioeconomic level was measured using the Spanish Deprivation Index by census tract. We produced sex- and age-specific life expectancies at birth by quintiles of deprivation, and life tables by census tract and province. Life expectancy at birth was higher among women than among men. Women and men in the most deprived census tracts in Spain lived 3.2 and 3.8 years less than their counterparts in the least deprived areas. A higher life expectancy in the northern regions of Spain was discovered. Life expectancy was higher in provincial capitals than in rural areas. We found a significant life expectancy gap and geographical variation by sex and socioeconomic status in Spain. The gap was more pronounced among men than among women. Understanding the association between life expectancy and socioeconomic status could help in developing appropriate public health programs. Furthermore, the life tables we produced are needed to estimate cancer specific survival measures by socioeconomic status. Therefore, they are important for cancer control in Spain.


Biomarker modulation following short-term vorinostat in women with newly diagnosed primary breast cancer.
Agents that target the epigenome show activity in breast cancer models. In preclinical studies, the histone deacetylase inhibitor vorinostat induces cell-cycle arrest, apoptosis, and differentiation. We evaluated biomarker modulation in breast cancer tissues obtained from women with newly diagnosed invasive disease who received vorinostat and those who did not. Tumor specimens were collected from 25 women who received up to 6 doses of oral vorinostat 300 mg twice daily and from 25 untreated controls in a nonrandomized study. Candidate gene expression was analyzed by reverse transcription PCR (RT-PCR) using the Oncotype DX 21-gene assay, and by immunohistochemistry for Ki-67 and cleaved caspase-3. Matched samples from treated women were analyzed for gene methylation by quantitative multiplex methylation-specific PCR (QM-MSP). Wilcoxon nonparametric tests were used to compare changes in quantitative gene expression levels pre- and post-vorinostat with changes in expression in untreated controls, and changes in gene methylation between pre- and post-vorinostat samples. Vorinostat was well tolerated and there were no study-related delays in treatment. Compared with untreated controls, there were statistically significant decreases in the expression of proliferation-associated genes Ki-67 (P = 0.003), STK15 (P = 0.005), and Cyclin B1 (P = 0.03) following vorinostat, but not in other genes by the Oncotype DX assay, or in expression of Ki-67 or cleaved caspase-3 by immunohistochemistry. Changes in methylation were not observed. Short-term vorinostat administration is associated with a significant decrease in expression of proliferation-associated genes in untreated breast cancers. This demonstration of biologic activity supports investigation of vorinostat in combination with other agents for the management of breast cancer.


Heterogeneity of fibrillin-rich microfibrils extracted from human skin of diverse ethnicity.
The dermal elastic fibre network is the primary effector of skin elasticity, enabling it to extend and recoil many times over the lifetime of the individual. Fibrillin-rich microfibrils (FRMs) constitute integral components of the elastic fibre network, with their distribution showing differential deposition in the papillary dermis across individuals of diverse skin ethnicity. Despite these differential findings in histological presentation, it is not known if skin ethnicity influences FRM ultrastructure. FRMs are evolutionarily highly conserved from jellyfish to man and, regardless of tissue type or species, isolated FRMs have a characteristic 'beads-on-a-string' ultrastructural appearance, with an average inter-bead distance (or periodicity) of 56 nm. Here, skin biopsies were obtained from the photoprotected buttock of healthy volunteers (18-27 years; African: n = 5; European: n = 5), and FRMs were isolated from the superficial papillary dermis and deeper reticular dermis and imaged by atomic force microscopy. In the reticular dermis, there was no significant difference in FRM ultrastructure between European and African participants. In contrast, in the more superficial papillary dermis, inter-bead periodicity was significantly larger for FRMs extracted from European participants than from African participants by 2.20 nm (p < .001). We next assessed whether these differences in FRM ultrastructure were present during early postnatal development by characterizing FRMs from full-thickness neonatal foreskin. Analysis of FRM periodicity identified no significant difference between neonatal cohorts (p = .865). These data suggest that at birth, FRMs are developmentally invariant. However, in adults of diverse skin ethnicity, there is a deviation in ultrastructure for the papillary dermal FRMs that may be acquired during the passage of time from child to adulthood. Understanding the mechanism by which this difference in papillary dermal FRMs arises warrants further study.


Neglected tropical diseases: diagnosis, clinical management, treatment and control.
Branded in 2005, "neglected tropical diseases" have gained traction in terms of advocacy, interest for research, enhanced funding and political will for their control and eventual elimination. Starting with an initial set of 13 neglected tropical diseases--seven helminth, three bacterial and three protozoal infections--the list considerably expanded to more than 40 diseases that now also includes viral, fungal and ectoparasitic infections. In this review, we provide a comprehensive overview of the neglected tropical diseases, their causative agents and the current geographical distribution, including their importance for the general practitioners seeing returning travellers and migrants in Switzerland. We characterise the most important of the neglected tropical diseases in terms of at-risk population, estimated number of infections, annual mortality rates and global burden, including current knowledge gaps. With an emphasis on neglected tropical diseases due to helminths, protozoa and ectoparasites, we review common diagnostic methods and current recommendations for treatment at the population level and the individual patient, thereby juxtaposing the situation in highly endemic countries on one side, with Switzerland on the other. We highlight the clinical presentation and management of the neglected tropical diseases in general and then elaborate on two examples, strongyloidiasis and leptospirosis. Our review provides a global perspective of neglected tropical diseases and we hope that it will prove useful for the general practitioner and clinician in Switzerland and elsewhere to enhance their suspicion index, differential diagnosis, clinical management and treatment, including referral to specialised clinics and laboratories when need be.


Specific CP110 Phosphorylation Sites Mediate Anaphase Catastrophe after CDK2 Inhibition: Evidence for Cooperation with USP33 Knockdown.
Chromosomal instability (CIN) is a hallmark of solid tumor biology and is implicated in carcinogenesis. Preferentially eliminating malignant cells by targeting CIN and aneuploidy is an attractive antineoplastic strategy. We previously reported that CDK2 antagonism causes lung cancer cells to undergo anaphase catastrophe and apoptosis through inhibition of phosphorylation of the centrosomal protein CP110. Cells with activating KRAS mutations were particularly sensitive to CDK2 inhibition due to downregulation of CP110 protein levels. This study investigated mechanisms of CDK2 antagonism that mediate anaphase catastrophe via changes in CP110 protein expression and how activated KRAS affects CP110 levels in lung cancers. Site-directed mutagenesis revealed candidate CDK phosphorylation sites of CP110 (residues Ser 170 and Thr 194) critical for conferring anaphase catastrophe by altering centrosome clustering in mitosis. Intriguingly, KRAS mutation can promote CP110 protein degradation by upregulating the ubiquitin ligase SCF(cyclinF), which targets CP110 protein for destabilization. Finally, CDK2 inhibitor response was enhanced when combined with knockdown of the deubiquitinase USP33 that in turn accelerates CP110 protein degradation. Thus, this study provides molecular pharmacologic insights into how CP110 expression regulates response to CDK2 inhibition. An improved understanding of in vitro antineoplastic mechanisms of combining CDK2 antagonism with induced CP110 repression provides a rationale for exploring clinical consequences of this strategy. Taken together, preclinical findings obtained from combining CDK2 inhibition with USP33 repression have implications for treating patients with non-small cell lung cancers.


Decreases in diabetes-free life expectancy in the U.S. and the role of obesity.
With increasing life expectancy in the U.S., it is important to know whether a longer life expectancy means a longer healthy life span or a prolonged period of later-life morbidity. This study examines changes in lifetime without diabetes, a leading cause of morbidity in later life. Using demographic methods and nationally representative data, we estimated changes in diabetes-free life expectancy between 1980-1989 and 2000-2004 for adult men and women in the U.S., estimated the contribution of changes in age-specific diabetes rates, and examined the changing effects of weight status on diabetes risks. While life expectancy at age 18 for men and women increased between the 1980s and the 2000s, diabetes-free life expectancy at age 18 decreased by 1.7 years for men and 1.5 years for women. The proportion of 18-year-olds who would develop diabetes in their lifetimes increased by almost 50% among women and almost doubled among men. Obese individuals experienced the greatest losses in diabetes-free life expectancy during this period, estimated at 5.6 years for men and 2.5 years for women. Diabetes-free life expectancy decreased for both men and women between 1980-1989 and 2000-2004, and these decreases are almost entirely attributable to large increases in diabetes incidence among obese individuals.


Trained Immunity and Reactivity of Macrophages and Endothelial Cells.
Innate immune cells can develop exacerbated immunologic response and long-term inflammatory phenotype following brief exposure to endogenous or exogenous insults, which leads to an altered response towards a second challenge after the return to a nonactivated state. This phenomenon is known as trained immunity (TI). TI is not only important for host defense and vaccine response but also for chronic inflammations such as cardiovascular and metabolic diseases such as atherosclerosis. TI can occur in innate immune cells such as monocytes/macrophages, natural killer cells, endothelial cells (ECs), and nonimmune cells, such as fibroblast. In this brief review, we analyze the significance of TI in ECs, which are also considered as innate immune cells in addition to macrophages. TI can be induced by a variety of stimuli, including lipopolysaccharides, BCG (bacillus Calmette-Guerin), and oxLDL (oxidized low-density lipoprotein), which are defined as risk factors for cardiovascular and metabolic diseases. Furthermore, TI in ECs is functional for inflammation effectiveness and transition to chronic inflammation. Rewiring of cellular metabolism of the trained cells takes place during induction of TI, including increased glycolysis, glutaminolysis, increased accumulation of tricarboxylic acid cycle metabolites and acetyl-coenzyme A production, as well as increased mevalonate synthesis. Subsequently, this leads to epigenetic remodeling, resulting in important changes in chromatin architecture that enables increased gene transcription and enhanced proinflammatory immune response. However, TI pathways and inflammatory pathways are separated to ensure memory stays when inflammation undergoes resolution. Additionally, reactive oxygen species play context-dependent roles in TI. Therefore, TI plays significant roles in EC and macrophage pathology and chronic inflammation. However, further characterization of TI in ECs and macrophages would provide novel insights into cardiovascular disease pathogenesis and new therapeutic targets. Graphic Abstract: A graphic abstract is available for this article.


[Multicenter study on the prevention of congenital toxoplasmosis in Buenos Aires].
Toxoplasmosis is an infection caused by Toxoplasma gondii, an intracellular parasite of universal distribution, with a variable prevalence depending on the region. This infection causes both morbidity and mortality in the fetus and newborn. The present study is an evaluation of the Argentine Consensus Guidelines regarding prenatal prevention of toxoplasmosis. Screening tests in pregnant women were done in nine different hospitals within the city of Buenos Aires and surroundings, where 19825 births between May 1st 2006 and April 30th 2007 were registered. Screening tests were done in 13632 pregnant women, using IgG and IgM determinations by ELISA. If acute infection was suspected, the patient's serum was sent to the reference laboratory to fulfill the pending tests: Sabin Feldman, ISAGA M, ISAGA A, ISAGA E and avidity. Clinical and serologic evaluation was done to all newborn of these mothers. Three hundred and fifty one specimens were sent and analyzed. Conclusions from the analysis were as follows: 121 (32%) patients probably acquired the infection during pregnancy, in 176 (46%) patients, acute infection was excluded, in 37 women (10%) serologic results were inconclusive, and in 47 (12%) the interpretation of results was impossible due to lack of information on the exact gestational age. Clinical and serologic control was performed in 94 newborns of mothers infected during pregnancy, and 5 congenital toxoplasmosis were detected, with fetal damage, four corioretinitis and one case of microcephaly. This study allowed us to validate the Argentine Consensus of Congenital Toxoplasmosis Guidelines.


Computer simulation study on the self-assembly of unimodal and bimodal polymer-grafted nanoparticles in a polymer melt.
The controllable distribution of nanoparticles (NPs) in polymer nanocomposites (PNCs) is a challenge in materials science. An important method is grafting chains that are chemically identical to the polymer matrix on NPs. By performing comprehensive molecular dynamics simulations, the self-assembly behavior of polymer-grafted NPs in a polymer matrix is investigated in this study. The relationship between the grafted chain length N, grafting density σ and the NPs' self-assembly morphologies is studied. Phase diagrams of the NP self-assembly structures for both unimodal and bimodal grafted NP systems are constructed on a parameter space, where P is the matrix polymer chain length. NP self-assembly structures of strings, connected/sheet and small clusters are identified in different regions. In order to quantitatively characterize the NP self-assembly morphology, we define a morphological measurement parameter which characterizes the distribution of the Voronoi cell volume of the NPs. Using this parameter, we discuss the influences of both long and short grafted chains on the dispersion of bimodal polymer-grafted NPs in a polymer melt. We find that the short grafted chains can not only shield the NP surface from the polymer matrix but also elongate the long grafted chains into the polymer matrix, therefore favoring a better dispersion of NPs. Our results also indicate that the bimodal grafted NPs will not be fully dispersed until the short grafted chains are dense enough to elongate the long grafted chains, hence forming a wetting NP/matrix interface.


A review of bartonellosis in Ecuador and Colombia.
A review of the literature regarding bartonellosis or Carrion's disease in Colombia and Ecuador is presented, together with observations made by the author in areas of both countries from which the disease has been recorded. There is evidence from pre-Columbian artifacts that verruga peruana, the cutaneous form of the disease, was present in Ecuador at least 1,000 years prior to the arrival of Europeans. These artifacts were discovered in the coastal province of Manabi, a low-lying area very different from the high Andean valleys of Peru with which bartonellosis is normally associated. Most of the cases recorded in recent years from this coastal area. The disease does not appear to have occurred in Colombia before the 1930s and only one case has been reported during the past 40 years. The possibility of many more subclinical cases being present in both Ecuador and Colombia is discussed, together with the possibility that the acquired immunodeficiency syndrome epidemic will reveal a higher prevalence among the inhabitants of endemic areas than previously suspected. Although the suspected vector of Bartonella bacilliformis, the sand fly Lutzomyia verrucarum, has not been recorded from Ecuador or Colombia, related species are present in endemic areas and may be involved in transmission.


Comparison of brain natriuretic peptide (BNP) and NT-proBNP in screening for pulmonary arterial hypertension in patients with systemic sclerosis.
To compare the performance of brain natriuretic peptide (BNP) and N-terminal pro-brain natriuretic peptide (NT-proBNP) in screening for pulmonary arterial hypertension (PAH) in systemic sclerosis (SSc). Between January 2008 and March 2009, outpatients referred to our unit and satisfying LeRoy criteria for SSc were assessed for PAH. Doppler echocardiography, BNP measurement, and NT-proBNP measurement were done concomitantly for a complete clinical, instrumental, and biochemical evaluation. Right-heart catheterization was carried out in cases of suspected PAH [estimated pulmonary arterial pressure (PAP) ≥ 36 mm Hg; diffusion capacity for carbon monoxide (DLCO) ≤ 50% of predicted value; 1-year DLCO decline ≥ 20% in absence of pulmonary fibrosis; unexplained dyspnea]. One hundred thirty-five patients were enrolled (124 women, 11 men; 96 limited SSc, 39 diffuse SSc); precapillary PAH was found in 20 patients (15 limited SSc, 5 diffuse SSc). The estimated PAP correlated with both BNP (R = 0.3; 95% CI 0.14-0.44) and NT-proBNP (R = 0.3, 95% CI 0.14-0.45). BNP [area under the curve (AUC) 0.74, 95% CI 0.59-0.89] was slightly superior to NT-proBNP (AUC 0.63, 95% CI 0.46-0.80) in identification of PAH, with diagnosis cutoff values of 64 pg/ml (sensitivity 60%, specificity 87%) and 239.4 pg/ml (sensitivity 45%, specificity 90%), respectively. BNP (log-transformed, p = 0.032) and creatinine (p = 0.049) were independent predictors of PAH, while NT-proBNP was not (p = 0.50). In our single-center study, the performance of BNP was slightly superior to that of NT-proBNP in PAH screening of patients with SSc, although normal levels of these markers do not exclude diagnosis. We observed that impaired renal function is associated with an increased risk of PAH in SSc. Further multicenter studies are needed to confirm our results (ClinicalTrials.gov ID NCT00617487).


Glutamate-Weighted Chemical Exchange Saturation Transfer Magnetic Resonance Imaging Detects Glutaminase Inhibition in a Mouse Model of Triple-Negative Breast Cancer.
Glutamate is an important metabolite of glutaminolysis, a metabolic pathway used by many aggressive cancers, including triple-negative breast cancer (TNBC). With the exception of the brain, in vivo detection of glutamate in tissues using 1H magnetic resonance spectroscopy (MRS) is challenging. Compared with MRS, glutamate-weighted chemical exchange saturation transfer MR imaging (GluCEST MRI) offers a more sensitive detection mechanism that is free of glutamine interference. Here, we developed a robust, highly repeatable GluCEST MRI protocol in mice bearing human TNBC xenografts and treated with a potent glutaminase inhibitor, CB-839. In paired studies, treatment with CB-839 for 2 days reduced the GluCEST asymmetry value compared with baseline (P < 0.05, n = 10). The absolute change of the GluCEST asymmetry value was -2.5 percent points after CB-839 treatment versus +0.3 after vehicle (P < 0.01). Correspondingly, treatment with CB-839 reduced tumor glutamate concentrations by 1.5 mmol/L, consistent with prior calibration between changes of the GluCEST value versus tissue glutamate concentration; CB-839, however, did not change tumor intracellular pH. These results demonstrate in a mouse model of breast cancer the utility of GluCEST MRI to detect the early response to glutaminase inhibition.Significance: A sensitive method enables noninvasive detection of tumor response to inhibitors of glutamine metabolism. Cancer Res; 78(19); 5521-6. ©2018 AACR.


Quantitative H2[(15)O]-PET in Pediatric Moyamoya Disease: Evaluating Perfusion before and after Cerebral Revascularization.
Moyamoya disease (MMD) is an idiopathic intracranial angiopathy with a progressive spontaneous occlusion of the circle of Willis resulting in repeated ischemia if not diagnosed and treated early, especially in children. Prevention of stroke is achieved by revascularization of the affected cerebral regions. Functional imaging techniques such as H2[(15)O]-Positron emission tomography (PET) allow quantification of cerebral perfusion/blood flow (CBF) and in particular cerebrovascular response after acetazolamide (AZA) challenge. The cerebrovascular reserve (CVR) can then be calculated and used to identify regions at risk of infarct, hence allowing surgery to be specifically targeted and personalized. Pediatric patients with diagnosed MMD underwent initial H2[(15)O]-PET scans at baseline and after stimulation with AZA. Indication for surgery was then based collectively on the extent of disease observed clinically and on magnetic resonance imaging, on the arterial territories involved, as seen in angiography and the respective regional CVR observed in PET. Cerebral revascularization surgeries were subsequently performed, tailored to the individual patient. Postoperative assessment of clinical outcome was augmented with follow-up PET (median duration after surgery, 10.4 months). CBF at baseline, after AZA and CVR were compared between presurgery and postsurgery scans in the areas supplied by the major cerebral arteries. Parametric images reflecting CBF, response to AZA and CVR clearly showed deficits in cortical but not subcortical regions or cerebellum. AZA-CBF and CVR deficits were most clear in middle cerebral artery and anterior cerebral artery (ACA) regions. In addition to the clinical symptomatology, angiography, AZA-CBF, and CVR images allowed the laterality of deficits to be clearly visualized for tailored surgery and the indication for targeted ACA or posterior cerebral artery revascularization to be assessed. Comparison of baseline CBF, AZA-CBF, and CVR between presurgery and postsurgery scans in revascularized areas revealed a significant improvement in baseline and AZA-CBF after surgery. Although no significant differences in CVR after revascularization surgery were found, a clear improvement of the deficits apparent in AZA-CBF in revascularized regions was found. We demonstrate that quantitative H2[(15)O]-PET is a highly useful tool to direct surgical intervention in MMD. Detailed quantitative analysis of CBF changes and CVR after surgery supports a targeted surgical approach.


Mercury and other mining-related contaminants in ospreys along the Upper Clark Fork River, Montana, USA.
We investigated links between mining-related contaminants in river sediment and their occurrence in nestling ospreys (Pandion haliaetus) in the Clark Fork River Basin, Montana, USA. Blood and feather samples from 111 osprey chicks were collected during 4 years from nests along river sections with greatly different sediment concentrations of arsenic (As), cadmium (Cd), copper (Cu), lead (Pb), zinc (Zn), and mercury (Hg). No significant differences between river sections were found among Zn (3,150 ± 160 μg L(-1)) and Cd (<5 μg L(-1)) concentrations in blood. Cu, Pb, and As concentrations in blood were significantly increased in chicks from the most contaminated river sections (mean values of 298, 8.9, and 100 μg L(-1), respectively). Cu, Zn, and Pb concentrations increased significantly during a year of above-average river runoff combined with high suspended sediment loads in rivers. Total Hg concentrations in blood and feathers were highly correlated and depended on the geographic locations of the nests. The lowest blood concentrations of Hg were observed in the most upstream river section (mean 151 μg L(-1)) where total sediment concentrations were increased (0.80 mg kg(-1)). River sections with intermediate blood concentrations (mean 206 and 303 μg L(-1)) were associated with low to intermediate sediment concentrations (0.058 and 0.46 mg kg(-1)). The highest concentrations of Hg in ospreys (mean 548 μg L(-1)) were observed downstream from a contaminated tributary (1-4 mg kg(-1) in sediment). In river sections with lower Hg concentrations in sediment, there was a negative correlation between blood Hg concentration and chick mass, presumably due to high deposition rates into growing feathers. This relationship was absent in sections of high Hg exposure. Osprey blood and feathers are suitable for monitoring Hg in aquatic ecosystems; however, responses of As, Cd, Cu, Pb, and Zn are more subtle.