Implementation of a Multiplex and Quantitative Proteomics Platform for Assessing Protein Lysates Using DNA-Barcoded Antibodies.
Molecular analysis of tumors forms the basis for personalized cancer medicine and increasingly guides patient selection for targeted therapy. Future opportunities for personalized medicine are highlighted by the measurement of protein expression levels via immunohistochemistry, protein arrays, and other approaches; however, sample type, sample quantity, batch effects, and "time to result" are limiting factors for clinical application. Here, we present a development pipeline for a novel multiplexed DNA-labeled antibody platform which digitally quantifies protein expression from lysate samples. We implemented a rigorous validation process for each antibody and show that the platform is amenable to multiple protocols covering nitrocellulose and plate-based methods. Results are highly reproducible across technical and biological replicates, and there are no observed "batch effects" which are common for most multiplex molecular assays. Tests from basal and perturbed cancer cell lines indicate that this platform is comparable to orthogonal proteomic assays such as Reverse-Phase Protein Array, and applicable to measuring the pharmacodynamic effects of clinically-relevant cancer therapeutics. Furthermore, we demonstrate the potential clinical utility of the platform with protein profiling from breast cancer patient samples to identify molecular subtypes. Together, these findings highlight the potential of this platform for enhancing our understanding of cancer biology in a clinical translation setting.


Waist to hip ratio and psychosocial factors in adults with insulin-dependent diabetes mellitus: the Pittsburgh Epidemiology of Diabetes Complications study.
The relationship between waist to hip ratio (WHR) and psychosocial factors has seldom been investigated, although both may contribute to cardiovascular risk. Therefore, these variables were examined in adults with insulin-dependent diabetes mellitus ([IDDM] N = 592; mean age, 29 years; mean duration, 20 years), a population at increased risk of developing cardiovascular disease. Moreover, the association between changes in psychosocial factors and change in WHR was considered. After adjusting for body mass index (BMI), WHR in men was correlated with higher levels of depressive symptomatology (r = .19, P < .001), greater anxiety (r = .13, P < .05), less social support (r = -.20, P < .01), and lower type A scores (r = -.25, P < .001). In women, WHR was significantly correlated with higher levels of depressive symptomatology (r = .18, P < .01), greater stress (r = .16, P < .01), and alcohol consumption (r = .12, P < .05). For both sexes, smokers had a significantly greater mean WHR than nonsmokers (P < .01). For men, multiple regression analyses adjusting for BMI and age demonstrated that smoking, lower income, less exercise, and lower type A scores were the most significant variables associated with WHR. In women, the independent predictors of WHR were a history of smoking, lower educational level, and depressive symptomatology. The most significant independent predictors of change in WHR from baseline to 2-year follow-up study were change in weight (men), change in BMI (women), and change in depression scores (both sexes). These results suggest that psychosocial factors may affect cardiovascular disease risk through their influence on body fat distribution, and both may be important in identifying those most at risk for cardiovascular disease in populations with IDDM.


Sulodexide for treating venous leg ulcers.
Venous leg ulcers are common, chronic wounds caused by venous diseases, with a high recurrence rate and heavy disease burden. Compression therapy (bandages or stockings) is the first choice treatment for venous leg ulcers. However, when ulcers remain unhealed, medication can also be used with or without compression therapy. Sulodexide, a highly purified glycosaminoglycan (a naturally occurring molecule) has antithrombotic and profibrinolytic properties (it reduces the formation of blood clots) as well as anti-inflammatory effects. Sulodexide has been studied as a potential treatment for venous leg ulcers. To assess the efficacy and safety of sulodexide for treating venous leg ulcers. In July 2015 we searched: The Cochrane Wounds Specialised Register; The Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library); Ovid MEDLINE; Ovid MEDLINE (In-Process & Other Non-Indexed Citations); Ovid EMBASE; EBSCO CINAHL; Chinese Biomedical Literature Database (CBM); China National Knowledge Infrastructure Database (CNKI); Wan Fang and VIP. We also searched clinical trials registries to identify ongoing studies, as well as references listed in relevant publications. There were no restrictions based on date of publication, language or study setting. Randomised controlled trials (RCTs) involving people with a diagnosis of venous leg ulcers which compared sulodexide with placebo or any other drug therapy (such as pentoxifylline, flavonoids, aspirin), with or without compression therapy. We used standard Cochrane methodological procedures. The authors independently selected studies, extracted data and assessed risk of bias. We pooled data to present the risk ratio (RR) with 95% confidence interval (CI), or presented a narrative summary. We assessed overall evidence quality according to the GRADE approach. We included four RCTs with a total of 463 participants (aged 42 years to 93 years); one report was only available as a published abstract.Meta-analysis of three RCTs suggests an increase in the proportion of ulcers completely healed with sulodexide as an adjuvant to local treatment (including wound care and compression therapy) compared with local treatment alone (rate of complete healing with sulodexide 49.4% compared with 29.8% with local treatment alone; RR 1.66; 95% CI 1.30 to 2.12). This evidence for sulodexide increasing the rate of complete healing is low quality due to risk of bias. It is unclear whether sulodexide is associated with any increase in adverse events (4.4% with sulodexide versus 3.1% with no sulodexide; RR 1.44; 95% CI 0.48 to 4.34). The evidence for adverse events is very low quality, downgraded twice for risk of bias and once for imprecision. Sulodexide may increase the healing of venous ulcers, when used alongside local wound care, however the evidence is only low quality and the conclusion is likely to be affected by new research. It is not clear whether sulodexide is associated with adverse effects. The standard dosage, route and frequency of sulodexide reported in the trials was unclear. Further rigorous, adequately powered RCTs examining the effects of sulodexide on healing, ulcer recurrence, quality of life and costs are necessary.


The Wellcome Prize Lecture. A map of auditory space in the mammalian brain: neural computation and development.
The experiments described in this review have demonstrated that the SC contains a two-dimensional map of auditory space, which is synthesized within the brain using a combination of monaural and binaural localization cues. There is also an adaptive fusion of auditory and visual space in this midbrain nucleus, providing for a common access to the motor pathways that control orientation behaviour. This necessitates a highly plastic relationship between the visual and auditory systems, both during postnatal development and in adult life. Because of the independent mobility of difference sense organs, gating mechanisms are incorporated into the auditory representation to provide up-to-date information about the spatial orientation of the eyes and ears. The SC therefore provides a valuable model system for studying a number of important issues in brain function, including the neural coding of sound location, the co-ordination of spatial information between different sensory systems, and the integration of sensory signals with motor outputs.


Immune Stimulation Using a Gut Microbe-Based Immunotherapy Reduces Disease Pathology and Improves Barrier Function in Ulcerative Colitis.
Background: Current ulcerative colitis (UC) treatments are focused on symptom management primarily via immune suppression. Despite the current arsenal of immunosuppressant treatments, the majority of patients with UC still experience disease progression. Importantly, aggressive long-term inhibition of immune function comes with consequent risk, such as serious infections and malignancy. There is thus a recognized need for new, safe and effective treatment strategies for people living with UC that work upstream of managing the symptoms of the disease. The objective of this study was to evaluate a microbial-based treatment, QBECO, that functions to productively activate rather than suppress mucosal immune function as a novel approach to treat UC. Methods: Two established models of experimental colitis, namely chemically-induced DSS colitis and the spontaneous colitis that develops in Muc2 deficient mice, were used to assess whether QBECO treatment could ameliorate gastrointestinal disease. A small exploratory 16-week QBECO open-label trial was subsequently conducted to test the safety and tolerability of this approach and also to determine whether similar improvements in clinical disease and histopathology could be demonstrated in patients with moderate-to-severe UC. Results: QBECO treatment successfully reduced inflammation and promoted mucosal and histological healing in both experimental models and in UC patients. The preclinical models of colitis showed that QBECO ameliorated mucosal pathology, in part by reducing inflammatory cell infiltration, primarily that induced by neutrophils and inflammatory T cells. The most rapid and noticeable change observed in QBECO treated UC patients was a marked reduction in rectal bleeding. Conclusion: Collectively, this work demonstrates for the first time that strategically activating immune function rather than suppressing it, not only does not worsen colitis induced-damage, but may lead to an objective reduction in UC disease pathology.


Missing heritability: is the gap closing? An analysis of 32 complex traits in the Lifelines Cohort Study.
Despite the recent explosive rise in number of genetic markers for complex disease traits identified in genome-wide association studies, there is still a large gap between the known heritability of these traits and the part explained by these markers. To gauge whether this 'heritability gap' is closing, we first identified genome-wide significant SNPs from the literature and performed replication analyses for 32 highly relevant traits from five broad disease areas in 13 436 subjects of the Lifelines Cohort. Next, we calculated the variance explained by multi-SNP genetic risk scores (GRSs) for each trait, and compared it to their broad- and narrow-sense heritabilities captured by all common SNPs. The majority of all previously-associated SNPs (median=75%) were significantly associated with their respective traits. All GRSs were significant, with unweighted GRSs generally explaining less phenotypic variance than weighted GRSs, for which the explained variance was highest for height (15.5%) and varied between 0.02 and 6.7% for the other traits. Broad-sense common-SNP heritability estimates were significant for all traits, with the additive effect of common SNPs explaining 48.9% of the variance for height and between 5.6 and 39.2% for the other traits. Dominance effects were uniformly small (0-1.5%) and not significant. On average, the variance explained by the weighted GRSs accounted for only 10.7% of the common-SNP heritability of the 32 traits. These results indicate that GRSs may not yet be ready for accurate personalized prediction of complex disease traits limiting widespread adoption in clinical practice.


Genic variation in abundant soluble proteins of Drosophila melanogaster and Drosophila pseudoobscura.
Genic variation was surveyed for 20 proteins of Drosophila melanogaster and 18 proteins of D. pseudoobscura. Analysis was by extraction and one-dimensional polyacrylamide gel electrophoresis under nondenaturing conditions, followed by staining with Coomassie Brilliant Blue to detect soluble proteins present in relatively large amounts ("abundant soluble proteins"). D. melanogaster was polymorphic for 65% of its protein loci and an individual was heterozygous for 10% of its loci. The respective figures for D pseudoobscura were 61% and 11%. These estimates of genic variation fall between previously published estimates obtained for these species by one-dimensional electrophoresis of soluble enzymes and those obtained by two-dimensional electrophoresis of solubilized abundant proteins. However, variation for both species could be strongly partitioned between loci, on the basis of tissue and stage expression of the proteins. The results are discussed with respect to their bearing on the possibility that abundant proteins constitute a distinct class of proteins less polymorphic than soluble enzymes.


Reproductive endocrinology of the explosively breeding desert spadefoot toad, Scaphiopus couchii.
The spadefoot toad, Scaphiopus couchii, is an explosively breeding, desert dwelling amphibian that shows two characteristics hypothesized to select for a dissociated breeding pattern: it lives in a harsh environment and it has a very short, but predictable breeding period. We tested the hypothesis that these factors select for a dissociated breeding cycle by measuring plasma steroid hormones and the gametogenic cycles in a free-living population of S. couchii. Blood and tissue samples were obtained from prebreeding, calling (males only), amplexed, and postbreeding toads. In males, plasma testosterone (T), dihydrotestosterone (DHT), and corticosterone (B) differed significantly among reproductive groups, with calling and amplexing toads having the highest levels of each of these steroids. In females, plasma T, DHT, B, progesterone (P), and estradiol (E2) were measured; all steroids except B changed significantly among mating groups. Plasma T and E2 were greatest in prebreeding and amplexed females, while P and DHT were highest only in amplexed females. Primary spermatogonia (SPG I), secondary spermatogonia (SPG II), spermatocytes, spermatids (SPT), and spermatozoa embedded in Sertoli cells (SPS) all changed significantly among male breeding groups. Although all stages were present in all breeding groups, early stages of spermatozoa (SPG I and SPG II) were greatest in postbreeding males, while late stages (SPT and SPS) were highest in prebreeding, calling, and amplexing males. In females, oocyte stages I, II, III, V, and VI and atretic follicles also differed significantly among mating groups. Prebreeding female ovaries were dominated by late-stage (V and VI) oocytes which were subsequently ovulated during amplexus. In postbreeding females, oogenesis was well under way as indicated by the rising numbers of stage II and III oocytes. These results indicate that aspects of gametogenesis occur during and after breeding in both sexes. Although the results demonstrate that gonadal activity was high during the period of sexual activity, suggesting that S. couchii most closely fits an associated pattern of reproduction as defined by Crews (1984, Horm. Behav. 18, 22-28), we also found that gonadal activity continued past the breeding season. For this reason, S. couchii does not completely conform to an associated pattern.


Strategic Combinations: The Future of Oncolytic Virotherapy with Reovirus.
The dominant cancer treatment modalities such as chemotherapy, radiotherapy, and even targeted kinase inhibitors and mAbs are limited by low efficacy, toxicity, and treatment-resistant tumor subclones. Oncolytic viral therapy offers a novel therapeutic strategy that has the potential to dramatically improve clinical outcomes. Reovirus, a double-stranded benign human RNA virus, is a leading candidate for therapeutic development and currently in phase III trials. Reovirus selectively targets transformed cells with activated Ras signaling pathways; Ras genes are some of the most frequently mutated oncogenes in human cancer and it is estimated that at least 30% of all human tumors exhibit aberrant Ras signaling. By targeting Ras-activated cells, reovirus can directly lyse cancer cells, disrupt tumor immunosuppressive mechanisms, reestablish multicellular immune surveillance, and generate robust antitumor responses. Reovirus therapy is currently being tested in combination with radiotherapy, chemotherapy, immunotherapy, and surgery. In this review, we discuss the current successes of these combinatorial therapeutic strategies and emphasize the importance of prioritizing combination oncolytic viral therapy as reovirus-based treatments progress in clinical development. Mol Cancer Ther; 15(5); 767-73. ©2016 AACR.


Volumetric dilution, rather than sequestration best explains the low vitamin D status of obesity.
Vitamin D status is known to be poor in obese individuals; there is no consensus as to the reason. Cross-sectional study of the relation between serum 25-hydroxyvitamin D (25(OH)D) concentration and body size in the baseline data from unsupplemented adults entering two study cohorts in our research unit, N = 686. Regression analyses of body size variables against serum 25(OH)D concentration, using both linear and hyperbolic models. The fit to a hyperbolic model of 25(OH)D against body weight completely removed the obesity-related component of inter-individual variability in serum 25(OH)D concentration. The hyperbolic fit using total body weight was significantly better than any linear model, and specifically better than any using BMI. Dilution of ingested or cutaneously synthesized vitamin D in the large fat mass of obese patients fully explains their typically low vitamin D status. There is no evidence for sequestration of supplemental or endogenous cholecalciferol. Vitamin D replacement therapy needs to be adjusted for body size if desired serum 25(OH)D concentrations are to be achieved.


Phenotypic rescue of a Drosophila model of mitochondrial ANT1 disease.
A point mutation in the Drosophila gene that codes for the major adult isoform of adenine nuclear translocase (ANT) represents a model for human diseases that are associated with ANT insufficiency [stress-sensitive B(1) (sesB(1))]. We characterized the organismal, bioenergetic and molecular phenotype of sesB(1) flies then tested strategies to compensate the mutant phenotype. In addition to developmental delay and mechanical-stress-induced seizures, sesB(1) flies have an impaired response to sound, defective male courtship, female sterility and curtailed lifespan. These phenotypes, excluding the latter two, are shared with the mitoribosomal protein S12 mutant, tko(25t). Mitochondria from sesB(1) adults showed a decreased respiratory control ratio and downregulation of cytochrome oxidase. sesB(1) adults exhibited ATP depletion, lactate accumulation and changes in gene expression that were consistent with a metabolic shift towards glycolysis, characterized by activation of lactate dehydrogenase and anaplerotic pathways. Females also showed downregulation of many genes that are required for oogenesis, and their eggs, although fertilized, failed to develop to the larval stages. The sesB(1) phenotypes of developmental delay and mechanical-stress-induced seizures were alleviated by an altered mitochondrial DNA background. Female sterility was substantially rescued by somatic expression of alternative oxidase (AOX) from the sea squirt Ciona intestinalis, whereas AOX did not alleviate developmental delay. Our findings illustrate the potential of different therapeutic strategies for ANT-linked diseases, based on alleviating metabolic stress.


Cyclosporine, low-density lipoprotein, and cholesterol.
Lipoproteins are known to be able to transport a variety of drugs. This report suggests that low-density lipoprotein not only functions as an important carrier of cyclosporine in plasma but also facilitates transport of cyclosporine across the cell membrane by means of the low-density lipoprotein receptor. Such a mechanism would explain (1) the similar tissue distribution of cyclosporine and the low-density lipoprotein receptor, (2) the increase in immunosuppression and toxicity with low total serum cholesterol levels, and (3) the relative absence of immunosuppression and toxicity with high levels of cyclosporine in the blood in patients with hypertriglyceridemia. In addition to receptor-mediated uptake, a disturbance of the blood-brain barrier is suggested as an explanation of the high frequency of cyclosporine-induced central nervous system toxicity after liver transplantation. Cyclosporine-induced inhibition of the mitochondrial steroid 26-hydroxylase, an enzyme involved in the formation of bile acids from cholesterol and deficient in patients with cerebrotendinous xanthomatosis, may cause or contribute to the observed central nervous system toxicity. It also may explain the similar clinical features of cyclosporine-induced central nervous system toxicity and cerebrotendinous xanthomatosis.


Characterization of fetal arrhythmias by means of fetal magnetocardiography in three cases of difficult ultrasonographic imaging.
Characterization of ultrasound detected fetal arrhythmias is generally performed by means of M-mode and pulsed Doppler echocardiography (fECHO), sonographic techniques that allow only indirect and approximate reconstruction of the true electrophysiological events that occur in the fetal heart. Several studies demonstrated the ability of fetal magnetocardiography (fMCG) to identify fetal arrhythmias. We report on three women, studied after the 32nd gestational week, who were referred for fMCG because of unsatisfying fetal cardiac visualization with fECHO due to maternal obesity, fetus in constant dorsal position hiding the fetal heart, intrauterine growth retardation, and oligohydramnios. Minor pericardial effusion was present in the third patient and digoxin therapy was given. FMCG were recorded with a 77-channel MCG system working in a shielded room. Independent Component Analysis (FastICA algorithm) was used to reconstruct fetal signals. The good quality of the retrieved fetal signals allowed real-time detection of arrhythmias and their classification as supraventricular extrasystoles (SVE), with/without aberrant ventricular conduction and/or atrioventricular block. The time course of the fetal cardiac rhythm was reconstructed for the entire recording duration; hence, fetal heart rate variability could be studied in time and frequency. Since isolated extrasystoles may progress to more hazardous supraventricular tachycardias, the noninvasive antenatal characterization of, even transient, fetal arrhythmias and their monitoring during pregnancy can be of great clinical impact.


Dipeptides of glutamate and aspartate may be endogenous neuroexcitants in the rat hippocampal slice.
The dipeptide N-acetylaspartylglutamate (NAAG), and possibly the related dipeptide aspartylglutamate (AG), have been found in high concentrations in rat brain, and have been shown to bind avidly and selectively to a subset of glutamate (GLUT) receptors. Certain observations regarding GLUT and aspartate might be explained if the endogenous transmitter were a compound composed of both amino acids. We therefore examined the electrophysiological actions of NAAG and AG in the rat in vitro rat hippocampal slice model. NAAG or AG and GLUT were applied locally to cells by a dual-barrel pressure technique. Intracellular recordings from 34 CA1 pyramidal neurons showed depolarizations and conductance increases resembling evoked excitatory postsynaptic potential in 15 of 20 cells exposed to NAAG, and 14 of 14 exposed to AG. Many GLUT-responsive sites did not respond to AG, and most did not respond to NAAG. Responses to NAAG were usually too small to induce cell firing; they were best detected, therefore, by intracellular recording. With extracellular unit recordings, GLUT was equally excitatory in stratum radiatum and pyramidale of CA1 (N = 19; p greater than 0.10, one-way analysis of variance). In contrast, AG was considerably more potent (N = 21; p less than 0.01) in stratum radiatum. NAAG was not noted to excite cells when applied to stratum pyramidale. The region of maximal responsiveness to AG in CA1 coincided with the area of the dendritic tree receiving Schaffer collateral-commissural afferents. This spatial profile, together with other neuropharmacological evidence, support the candidacy of glutamate-containing peptides as endogenous excitatory compounds in certain pathways of hippocampus.


Distinct immunomodulatory effects of a panel of nanomaterials in human dermal fibroblasts.
There are many efforts in understanding the effects of nanoparticles on cell viability and metabolism, however, not much is known regarding the distinct molecular mechanisms of inflammation and cellular stress using low dosing concentrations. To address this gap in the literature, we utilized a novel experimental design that specifically probes the effects of a panel of commonly studied engineered nanomaterials along immunomodulatory pathways, including NF-κB. The panel of particles selected for this study included quantum dot nanocrystals, titanium dioxide, hydroxylated fullerenes, and silver nanoparticles. Cell viability, antioxidant activity, select messenger RNA, and protein modulation were studied in primary human dermal fibroblasts (HDF) and NF-κB knockdown HDF cells. Inflammatory and non-inflammatory immune responses were measured using protein and real-time PCR array analysis from HDF cells exposed to sub-lethal concentrations of nanoparticles. Differences in cellular response to nanoparticles in protein and antioxidant experiments were evident in NF-κB knockdown cells. The methods used in the study, along with the resultant data sets, serve as a potential model for studying the complex pathway-specific biochemical responses in cell and tissue systems associated with nanoparticle exposures.


Three-dimensional reconstruction from serial sections in PC-Windows platform by using 3D_Viewer.
Three-dimensional (3D) reconstruction from serial sections allows identification of objects of interest in 3D and clarifies the relationship among these objects. 3D_Viewer, developed in our laboratory for this purpose, has four major functions: image alignment, movie frame production, movie viewing, and shift-overlay image generation. Color images captured from serial sections were aligned; then the contours of objects of interest were highlighted in a semi-automatic manner. These 2D images were then automatically stacked at different viewing angles, and their composite images on a projected plane were recorded by an image transform-shift-overlay technique. These composition images are used in the object-rotation movie show. The design considerations of the program and the procedures used for 3D reconstruction from serial sections are described. This program, with a digital image-capture system, a semi-automatic contours highlight method, and an automatic image transform-shift-overlay technique, greatly speeds up the reconstruction process. Since images generated by 3D_Viewer are in a general graphic format, data sharing with others is easy. 3D_Viewer is written in MS Visual Basic 6, obtainable from our laboratory on request.


[The complex etiopathogenesis of focal segmental glomerulosclerosis].
Focal segmental glomerulosclerosis (FSGS) is a pathological spectrum subtended by heterogeneous etiologies. A good knowledge of FSGS and its causes should be included in the nephrologists' clinical background, as it deeply influences the subsequent management of the affected patients. In fact, while immunosuppressive treatment should be considered in idiopathic FSGS, the treatment of secondary forms should primarily aim at curing or containing the underpinning etiologic factors. Furthermore, in contrast to secondary FSGS, idiopathic FSGS tends to relapse after kidney transplantation. Although FSGS has a wide spectrum of etiologies, several pathogenetic "moments" are shared. Furthermore, recent studies have identified a pool of glomerular cells potentially capable of regenerating lost podocytes; these cells might represent a promising therapeutic target. The primary aim of this review is to describe the etiologic factors associated with FSGS, with a focus on the main pathogenetic mechanisms involved in its development.


Are Advances in Survival Among the Oldest Old Seen Across the Spectrum of Health and Functioning?
Mortality rates have been reduced by half over the last 60 years for nonagenarians, and the progress is continuing. The greater survival might be due to overtreatment of severely physically and cognitively disabled individuals, which is a big concern for societies and individuals. The study population comprised two Danish birth cohorts: the 1905 Cohort and the 1915 Cohort. At age 95, all from the two cohorts who were still alive and living in Denmark were invited to participate in a health survey that used the same assessment instrument. A total of 2,670 (56.8%) persons participated in the two surveys and survival was assessed through a 7.3-year follow-up period during which 2,497 (93.5%) had died, and with virtually no loss to follow-up. Despite the increasing chance of surviving to age 95, the 1915 Cohort had significantly better health and functioning than the 1905 Cohort. The survival advantage in the 1915 Cohort continued in the follow-up period after age 95: Median survival length was 2.4 months longer, p = .011. This advantage was not statistically associated with different levels of activities of daily living, physical performance, cognitive functioning, self-rated health and life satisfaction. However, the advantage tended to be more pronounced among people with better health. Life span and health increases among the oldest old. The improvement in survival for 95-year olds born in 1915 compared with 1905 was seen across the whole spectrum of health and functioning, with a tendency towards bigger improvement among those in good health.


Effect of exercise-based management on multidirectional instability of the glenohumeral joint: a pilot randomised controlled trial protocol.
The most commonly recommended treatment for multidirectional instability (MDI) of the shoulder is exercise. Despite this recommendation, there is limited evidence to support the effectiveness of exercise. The aim of this paper is to describe a pilot randomised controlled trial comparing the effectiveness of 2 exercise programmes on outcomes of participants with MDI. Consenting participants between 12 and 35 years, with non-traumatic MDI will be randomly allocated to participate in either the Rockwood Instability programme or the Watson MDI programme. Both programmes involve 1 consultation per week for 12 weeks with a physiotherapist to prescribe and progress a home exercise programme. Outcomes will be assessed at baseline, 6, 12, 24 and 52 weeks. Primary outcome measures include the Melbourne Instability Shoulder Score and Western Ontario Shoulder Index. Secondary outcomes include scapular coordinates, scapular upward rotation angles, muscle strength, symptomatic onset, limiting factor and angle of limiting factor in abduction range, incidence of complete glenohumeral joint dislocation, global rating of change, satisfaction scores, the Orebro Musculoskeletal Pain Questionnaire, adverse events and compliance with the home exercise programme. Data will be analysed on intention-to-treat principles and a per protocol basis. This trial will evaluate whether there are differences in outcomes between the Rockwood and the Watson MDI programmes for participants with MDI. Participant confidentiality will be maintained with publication of results. Ethics approval: Faculty of Health Sciences (FHEC12/201). ACTRN12613001240730; Pre-results.


Brownian dynamics simulations of analytical ultracentrifugation experiments exhibiting hydrodynamic and thermodynamic non-ideality.
Hydrodynamic and thermodynamic non-ideality are important phenomena when studying concentrated and interacting systems in analytical ultracentrifugation (AUC). Here we present an extended Brownian Dynamics (BD) based algorithm which incorporates hydrodynamic and thermodynamic non-ideality. It can serve as an independent and versatile approach for the theoretical description of interparticulate interactions in AUC, as it allows tracking the trajectory of individual particles. Concentration dependencies of the sedimentation and diffusion coefficient have been implemented and validated for the extended BD model. For monodisperse systems, it is shown that profiles obtained by BD are in excellent agreement with well-established Lamm equation solvers. Moreover, important limits and restrictions of current Lamm equation based analysis methods are discussed. In particular, BD allows modeling and evaluation of AUC data of non-ideal polydisperse systems. This is relevant as most nanoparticulate systems are polydisperse in size. Here, a simulation for a polydisperse system including concentration effects is presented for the first time.


Assessment of 25-OH vitamin D levels and abnormal blood pressure response in female patients with cardiac syndrome X.
Vitamin D deficiency is associated with coronary artery disease, hypertension, heart failure, endothelial dysfunction, and metabolic syndrome. The pathophysiology of cardiac syndrome X (CSX) involves many pathways that are influenced by vitamin D levels. This study aimed to investigate the relationship between vitamin D deficiency and abnormal blood pressure response to exercise in patients with CSX. This was a cross-sectional and observational study. Fifty females with normal epicardial coronary arteries who presented with typical symptoms of rest or effort angina and 41 healthy age-matched female controls, were included. Patients with cardiomyopathy, severe valvular disease, congenital heart disease, and left ventricular hypertrophy were excluded. All patients underwent stress electrocardiography examination and 25-hydroxy (OH) vitamin D level measurements. Levels of 25-OH vitamin D were significantly lower in CSX patients (9.8±7.3 ng/mL vs. 18.1±7.9 ng/mL; p<0.001). Systolic blood pressure (SBP) (188±15 mm Hg vs. 179±17 mm Hg; p=0.013) and diastolic blood pressure (DBP) (98±9 mm Hg vs. 88±9 mm Hg; p<0.001) during peak exercise were higher in CSX patients. Levels of 25-OH vitamin D were negatively correlated with peak SBP (r=-0.310, p=0.004) and peak DBP (r=-0.535, p<0.001) during exercise. To discard the multicollinearity problem, two different models were used for multivariate analyses. In the first model, metabolic equivalents (METs) (p=0.003) and 25-OH vitamin D levels (p=0.001) were independent predictors. METs (p=0.007), 25-OH vitamin D levels (p=0.008), and peak DBP were determined as independent predictors in the second multivariate model. In patients with CSX, 25-OH vitamin D levels were lower than those in controls; moreover, 25-OH vitamin D deficiency was also associated with higher levels of peak DBP during exercise.


The utility of serology for elimination surveillance of trachoma.
Robust surveillance methods are needed for trachoma control and recrudescence monitoring, but existing methods have limitations. Here, we analyse data from nine trachoma-endemic populations and provide operational thresholds for interpretation of serological data in low-transmission and post-elimination settings. Analyses with sero-catalytic and antibody acquisition models provide insights into transmission history within each population. To accurately estimate sero-conversion rates (SCR) for trachoma in populations with high-seroprevalence in adults, the model accounts for secondary exposure to Chlamydia trachomatis due to urogenital infection. We estimate the population half-life of sero-reversion for anti-Pgp3 antibodies to be 26 (95% credible interval (CrI): 21-34) years. We show SCRs below 0.015 (95% confidence interval (CI): 0.0-0.049) per year correspond to a prevalence of trachomatous inflammation-follicular below 5%, the current threshold for elimination of active trachoma as a public health problem. As global trachoma prevalence declines, we may need cross-sectional serological survey data to inform programmatic decisions.


Tenascin-C Orchestrates an Immune-Suppressive Tumor Microenvironment in Oral Squamous Cell Carcinoma.
Inherent immune suppression represents a major challenge in the treatment of human cancer. The extracellular matrix molecule tenascin-C promotes cancer by multiple mechanisms, yet the roles of tenascin-C in tumor immunity are incompletely understood. Using a 4NQO-induced oral squamous cell carcinoma (OSCC) model with abundant and absent tenascin-C, we demonstrated that tenascin-C enforced an immune-suppressive lymphoid stroma via CCL21/CCR7 signaling, leading to increased metastatic tumors. Through TLR4, tenascin-C increased expression of CCR7 in CD11c+ myeloid cells. By inducing CCL21 in lymphatic endothelial cells via integrin α9β1 and binding to CCL21, tenascin-C immobilized CD11c+ cells in the stroma. Inversion of the lymph node-to-tumor CCL21 gradient, recruitment of T regulatory cells, high expression of anti-inflammatory cytokines, and matrisomal components were hallmarks of the tenascin-C-instructed lymphoid stroma. Ablation of tenascin-C or CCR7 blockade inhibited the lymphoid immune-suppressive stromal properties, reducing tumor growth, progression, and metastasis. Thus, targeting CCR7 could be relevant in human head and neck tumors, as high tenascin-C expression and an immune-suppressive stroma correlate to poor patient survival.


Quantitative analysis of motor unit action potentials in the subclavian muscle of healthy horses.
To evaluate the application of analysis of motor unit action potentials (MUAP) in horses and to obtain values of MUAP for the subclavian muscle of horses. 10 healthy adult Dutch Warmblood horses. Electromyographic examination of the subclavian muscle in conscious nonsedated horses was performed to evaluate insertional activity, spontaneous activity, MUAP variables, and recruitment patterns. Muscle and body temperatures were measured at the beginning and end of the procedure. Amplitude, duration, number of phases, and number of changes in direction (ie, turns) for all representative MUAP were analyzed to determine values for this muscle in this group of horses. Mean +/- SD duration of insertional activity was 471.7 +/- 33.45 milliseconds. Mean MUAP amplitude in the examined horses was 379 RV (95% confidence interval [CI], 349 to 410 microV). Mean MUAP duration of the subclavian muscle was 727 milliseconds (95% CI, 6.84 to 7.71 milliseconds). Mean number of phases was 2.9, and mean number of turns was 3.0. Prevalence of polyphasic MUAP defined as MUAP with > 4 phases, was 77%. Number of MUAP that had > 5 turns was 2.4%. Satellite potentials were found in 1.0% of the MUAP CONCLUSIONS AND CLINICAL RELEVANCE: This study revealed that electromyography including MUAP analysis can be performed in horses, and values for the subclavian muscle in healthy adult horses can be obtained. Analysis of MUAP could be a valuable diagnostic tool for use in discriminating between myogenic and neurogenic problems in horses.


Evidence That Phosphorylation of the α-Subunit of eIF2 Does Not Essentially Inhibit mRNA Translation in Wheat Germ Cell-Free System.
A mechanism based on reversible phosphorylation of the α-subunit of eukaryotic initiation factor 2 (eIF2α) has been confirmed as an important regulatory pathway for the inhibition of protein synthesis in mammalian and yeast cells, while plants constitute the significant exception. We studied the induction of TaeIF2α phosphorylation in germinated wheat (Triticum aestivum) embryos subjected to different adverse conditions. Data confirmed that formation of TaeIF2(αP) was not a general response, as no phosphorylation was observed under salt, oxidative, or heat stress. Nevertheless, treatment by salicylic acid, UV-light, cold shock and histidinol did induce phosphorylation of TaeIF2α of wheat as has been established previously for AteIF2α in Arabidopsis (Arabidopsis thaliana). The influence of TaeIF2α phosphorylation on translation of reporter mRNA with different 5'-untranslated regions (5'UTRs) was studied in wheat germ cell-free system (WG-CFS), in which TaeIF2α was first phosphorylated either by heterologous recombinant human protein kinase, HsPKR (activated by double-stranded (ds)RNA), or by endogenous protein kinase TaGCN2 (activated by histidinol). Pretreatment of WG-CFS with HsPKR in the presence of dsRNA or with histidinol resulted in intense phosphorylation of TaeIF2α; however, the translation levels of all tested mRNAs decreased by only 10-15% and remained relatively high. In addition, factor OceIF2 from rabbit (Oryctolagus cuniculus) bound GDP much more strongly than the homologous factor TaeIF2 from wheat germ. Furthermore, factor OceIF2B was able to stimulate guanine nucleotide exchange (GDP→GTP) on OceIF2 but had no effect on a similar exchange on TaeIF2. These results suggest that the mechanism of stress response via eIF2α phosphorylation is not identical in all eukaryotes, and further research is required to find and study in detail new plant-specific mechanisms that may inhibit overall protein synthesis in plants under stress.


Use of intervention strategies by assertive community treatment teams to promote patients' engagement.
This study explored the range of interventions and the use of more intrusive techniques by staff of assertive community treatment (ACT) teams to promote engagement, manage problem behaviors, and reinforce positive behaviors among patients. Individual and organizational characteristics that may be associated with these practices were identified. Between January and March 2006, clinicians (N=239) from 34 ACT teams participated in a one-time survey about their intervention strategies with patients, perceptions about the ACT team environment, and beliefs about persons with severe mental illness. Significant variation existed in the types of interventions employed across teams. The less intrusive strategies, including positive inducements and verbal guidance, were the most common. Other strategies that placed limits on patients but that were still considered less intrusive-such as medication monitoring and money management-were also common. Clinicians who reported working in more demoralized climates and having negative perceptions of mental illness were more likely to endorse leveraged or intrusive interventions. The findings of this study suggest significant variation across teams in the use of intervention strategies. Both perceptions of a demoralized organizational climate and stigmatizing beliefs about mental illness were correlated with the use of more intrusive intervention strategies. Future research on the role and appropriateness of more intrusive interventions in mental health treatment and the impact of such interventions on patient outcomes is warranted.


Role of Physical Therapists in the Management of Individuals at Risk for or Diagnosed With Venous Thromboembolism: Evidence-Based Clinical Practice Guideline.
The American Physical Therapy Association (APTA), in conjunction with the Cardiovascular & Pulmonary and Acute Care sections of APTA, have developed this clinical practice guideline to assist physical therapists in their decision-making process when treating patients at risk for venous thromboembolism (VTE) or diagnosed with a lower extremity deep vein thrombosis (LE DVT). No matter the practice setting, physical therapists work with patients who are at risk for or have a history of VTE. This document will guide physical therapist practice in the prevention of, screening for, and treatment of patients at risk for or diagnosed with LE DVT. Through a systematic review of published studies and a structured appraisal process, key action statements were written to guide the physical therapist. The evidence supporting each action was rated, and the strength of statement was determined. Clinical practice algorithms, based on the key action statements, were developed that can assist with clinical decision making. Physical therapists, along with other members of the health care team, should work to implement these key action statements to decrease the incidence of VTE, improve the diagnosis and acute management of LE DVT, and reduce the long-term complications of LE DVT.


Iron- and cobalt-catalyzed C(sp3)-H bond functionalization reactions and their application in organic synthesis.
Direct C-H bond functionalization catalyzed by non-precious transition metals is an attractive strategy in synthetic chemistry. Compared with the precious metals rhodium, palladium, ruthenium, and iridium commonly used in this field, catalysis based on non-precious metals, especially the earth-abundant ones, is appealing due to the increasing demand for environmentally benign and sustainable chemical processes. Herein, developments in iron- and cobalt-catalyzed C(sp3)-H bond functionalization reactions are described, with an emphasis on their applications in organic synthesis, i.e., the synthesis of natural products and pharmaceuticals and/or their modification.


Demonstration of link protein in proteoglycan aggregates from human intervertebral disc.
Proteoglycan aggregates free of non-aggregating proteoglycan have been prepared from the annuli fibrosi and nuclei pulposi of intervertebral discs of three human lumbar spines by extraction with 4M-guanidinium chloride, associative density gradient centrifugation, and chromatography on Sepharose CL-2B. The aggregate (A1-2B.V0) was subjected to dissociative density-gradient ultracentrifugation. Three proteins of Mr 38 900, 44 200 and 50 100 found in the fraction of low buoyant density (A1-2B.V0-D4) reacted with antibodies to link protein from newborn human articular cartilage. After reduction with mercaptoethanol, two proteins of Mr 43 000 and two of Mr 20 000 and 14 000 were seen. The A1-2B.V0-D4 fraction, labelled with 125I, coeluted with both hyaluronate and a hyaluronate oligosaccharide (HA14) on a Sepharose CL-2B column. HA10 and HA14 reduced the viscosity of A1 fractions; HA4, HA6 and HA8 did not. HA14 decreased the viscosity of disc proteoglycans less than it did that of bovine cartilage proteoglycans. Thus, although a link protein was present in human intervertebral disc, it stabilized proteoglycan aggregates less well than did the link protein from bovine nasal cartilage.


Gastro-protective effects by extracts of Petasites hybridus: the role of inhibition of peptido-leukotriene synthesis.
Extracts from Petasites hybridus are in therapeutic use for more than 2000 years. They have been claimed to improve gastrointestinal pain, lung-diseases such as asthma and cough, as well as spasms of the urogenital-tract. We have investigated these claims in animal models of gastro-intestinal ulcers and confirmed that alcoholic extracts of Petasites hybridus block the ethanol-induced gastric damage and reduce small intestinal ulcerations induced in rats by indomethacin. Searching for a possible mode of action, we found that ethanolic extracts of Petasites hybridus inhibit the peptido-leukotriene biosynthesis in mouse peritoneal macrophages but leave prostaglandin biosynthesis unimpaired. We conclude that the active principle could be useful in the treatment of gastro-intestinal human diseases and that Petasites hybridus may contribute to the inhibition of calcium-dependent processes such as leukotriene biosynthesis.


A controlled trial of including symptom data in computer-based care suggestions for managing patients with chronic heart failure.
Heart failure is common and associated with considerable morbidity and cost, yet physician adherence to treatment guidelines is suboptimal. We conducted a randomized controlled study to determine if adding symptom information to evidence-based, computer-generated care suggestions would affect treatment decisions among primary care physicians caring for outpatients with heart failure at two Veterans Affairs medical centers. Physicians were randomly assigned to receive either care suggestions generated with electronic medical record data and symptom data obtained from questionnaires mailed to patients within 2 weeks of scheduled outpatient visits (intervention group) or suggestions generated with electronic medical record data alone (control group). Patients had to have a diagnosis of heart failure and objective evidence of left ventricular systolic dysfunction. We assessed physician adherence to heart failure guidelines, as well as patients' New York Heart Association (NYHA) class, quality of life, satisfaction with care, hospitalizations, and outpatient visits, at 6 and 12 months after enrollment. Patients in the intervention (n = 355) and control (n = 365) groups were similar at baseline. At 12 months, there were no significant differences in adherence to care suggestions between physicians in the intervention and control groups (33% vs. 30%, P = 0.4). There were also no significant changes in NYHA class (P = 0.1) and quality-of-life measures (P >0.1), as well as no differences in the number of outpatient visits between intervention and control patients (6.7 vs. 7.1 visits, P = 0.48). Intervention patients were more satisfied with their physicians (P = 0.02) and primary care visit (P = 0.02), but had more all-cause hospitalizations at 6 months (1.5 vs. 0.7 hospitalizations, P = 0.0002) and 12 months (2.3 vs. 1.7 hospitalizations, P = 0.05). Adding symptom information to computer-generated care suggestions for patients with heart failure did not affect physician treatment decisions or improve patient outcomes.


The effect of exercise on pharmacokinetics and pharmacodynamics of levodopa.
The aim of our study was to evaluate the influence of low-intensity exercise on levodopa absorption and levodopa motor effect. We studied the pharmacokinetics and pharmacodynamics of levodopa under resting conditions and under a workload of 50 watts for 2 hours on a cycle ergometer in 12 parkinsonian patients with predictable fluctuations of motor disability. The patients attended the hospital on both days in a provoked off state. After a baseline assessment of motor disability using the Columbia University rating scale (CURS scale) and a blood test for measurement of the baseline levodopa concentration in the plasma, 100 mg levodopa and 25 mg benserazide were administered as a single dose orally. Blood samples for measurement of the levodopa concentration in the plasma were taken, and motor assessments were conducted at 15-minute intervals for 240 minutes and at 30-minute intervals from 240 to 360 minutes. All patients were able to perform the exercise program. The baseline Columbia University rating scale score did not differ significantly between both days. The mean levodopa concentration in plasma at half-maximal motor effect tended to be higher during exercise and indicated that the patients needed a higher levodopa concentration in plasma to achieve the half-maximal motor effect. The maximal levodopa concentration in plasma tended to be higher with exercise. Both trends did not reach statistical significance. In summary, there was not a negative or a positive net effect of exercise on pharmacokinetics and pharmacodynamics of levodopa. However, there were two counteracting trends: a trend toward better levodopa absorption and a trend toward a deteriorated concentration-effect correlation.


Distinguishing true from false positives in genomic studies: p values.
Distinguishing true from false positive findings is a major challenge in human genetic epidemiology. Several strategies have been devised to facilitate this, including the positive predictive value (PPV) and a set of epidemiological criteria, known as the "Venice" criteria. The PPV measures the probability of a true association, given a statistically significant finding, while the Venice criteria grade the credibility based on the amount of evidence, consistency of replication and protection from bias. A vast majority of journals use significance thresholds to identify the true positive findings. We studied the effect of p value thresholds on the PPV and used the PPV and Venice criteria to define usable thresholds of statistical significance. Theoretical and empirical analyses of data published on AlzGene show that at a nominal p value threshold of 0.05 most "positive" findings will turn out to be false if the prior probability of association is below 0.10 even if the statistical power of the study is higher than 0.80. However, in underpowered studies (0.25) with a low prior probability of 1 × 10(-3), a p value of 1 × 10(-5) yields a high PPV (>96 %). Here we have shown that the p value threshold of 1 × 10(-5) gives a very strong evidence of association in almost all studies. However, in the case of a very high prior probability of association (0.50) a p value threshold of 0.05 may be sufficient, while for studies with very low prior probability of association (1 × 10(-4); genome-wide association studies for instance) 1 × 10(-7) may serve as a useful threshold to declare significance.


Association of the HLA-DRB1 gene with premature death, particularly from cardiovascular disease, in patients with rheumatoid arthritis and inflammatory polyarthritis.
To examine the role of the variants of the PTPN22 and HLA-DRB1 genes as predictors of mortality in inflammatory polyarthritis (IP) and rheumatoid arthritis (RA). Patients were recruited from a primary care-based inception cohort of patients with IP and were followed up prospectively. For patients who died, the cause and date of death was obtained. Cox proportional hazards regression models were used to assess the association of the HLA-DRB1 (including the shared epitope [SE]) and PTPN22 genes with the risk of death from all causes and from cardiovascular disease (CVD) and to assess the interactions between SE, smoking, and anti-cyclic citrullinated peptide (anti-CCP) status, adjusted by age at symptom onset and sex. DNA samples were available from 1,022 IP patients. During followup, 751 of them (74%) satisfied the American College of Rheumatology 1987 criteria for RA, and 242 of them (24%) died. Carriage of 2 copies of SE alleles predicted death from all causes (hazard ratio [HR] 1.57 [95% confidence interval (95% CI) 1.1-2.2]) and from CVD (HR 1.68 [95% CI 1.1-2.7]). This effect was most marked for individuals with the HLA-DRB1*01/*04 combination. An interaction of smoking, SE alleles, and anti-CCP antibodies was observed and was associated with the greatest risk of death from CVD (HR 7.81 [95% CI 2.6-23.2]). No association of the PTPN22 gene with mortality was detected. SE alleles, particularly compound heterozygotes, are associated with death from all causes and from CVD, independently of autoantibody status. However, the combination of SE, smoking, and anti-CCP antibodies is associated with a high risk of premature death in patients with IP and RA, which raises the possibility of a targeted strategy to prevent CVD in these patients.


Effect of pendant group structure on the hydrolytic stability of polyaspartamide polymers under physiological conditions.
We describe the synthesis of metal chelating polymers based on polyaspartamide and polyglutamide backbones as carriers for (111)In in radioimmunoconjugates. These polymers [PAsp(DTPA), PGlu(DTPA)] have a biotin end group and diethylenetriaminepentaacetic acid (DTPA) chelators attached to the primary amines of the diethylenetriamine (DET) pendant groups of biotin-poly{N'-[N-(2-aminoethyl)-2-aminoethyl]aspartamide} [PAsp(DET)] and of biotin-poly{N'-[N-(2-aminoethyl)-2-aminoethyl]glutamide} [PGlu(DET)]. Like Asn-containing proteins and polypeptides, polyaspartamides undergo uncatalyzed degradation under model physiological conditions (10 mM phosphate buffer, pH 7.4, 150 mM NaCl). We studied the uncatalyzed degradation of the polyaspartamide polymers by size exclusion chromatography and found that the degradation rate was sensitive to the nature of the pendant groups. The metal-free polymer underwent somewhat slower degradation than the corresponding polymers in which the DTPA groups were saturated with Eu(3+) or In(3+), but even after 14 days, substantial fractions of the polymers survived. We conclude that these polymers undergo negligible degradation on the time scale (24-48 h) of radioimmunotherapy treatment of tumors with (111)In. From a mechanistic perspective, we note that these degradation rates are on the order of the deamidation rates reported [J. Peptide Res. 2004, 63, 426] for Asn-containing pentapeptides, with half-times on the order of 10 days, but much slower than the rapid decay (hours) reported recently [Biomaterials 2010, 31, 3707] for poly{N'-[N-(2-aminoethyl)-2-aminoethyl]aspartamide} itself. This variation in degradation rate can be explained in terms of the influence of positive charges on the pendant group enhancing the acidity of the side-chain amide nitrogen of the aspartamide repeat unit. The DET pendant group is positively charged at pH 7, but in indium-loaded PAsp(DTPA) this charge is offset by the net negative charge of the DTPA-In complex.


An auditory brainstem nucleus as a model system for neuronal metabolic demands.
The correlation between neuronal activity and metabolism is essential for coding, plasticity, neurological disorders and the interpretation of functional neuroimaging data. Most likely, metabolic requirements depend upon neuron type, and macroscopic energy demands vary with brain region. However, specific needs of individual neuron types are enigmatic. Therefore, we monitored metabolic activity in the lateral superior olive (LSO), an auditory brainstem nucleus containing only one neuron type. LSO neurons exhibit extreme but well-described biophysics with firing rates of several hundred hertz and low input resistances of a few megaohms. We recorded changes in NADH and flavin adenine dinucleotide (FAD) autofluorescence and O2 concentration in acute brainstem slices of Mongolian gerbils (Meriones unguiculatus) following electrical stimulation. The LSO shows the typical biphasic NADH/FAD response up to a physiologically relevant frequency of 400 Hz. In the same animal, we compared the LSO with the hippocampal CA1 region and the cerebral cortex. The rate of NADH/FADH2 consumption and regeneration was slowest in LSO. However, frequency dependence was only similar during the consumption phase but varied during regeneration within the three brain regions. Changes in NADH, FAD and O2 levels and blocking metabolic reactions indicate a pronounced contribution of mitochondrial oxidative phosphorylation in the LSO which is known for the other brain regions as well. Lactate transport and interconversion are involved in LSO metabolism as we found in immunohistochemical and pharmacological experiments. Our findings show that the LSO represents an apt, biophysically distinct model for brain metabolism and that neuronal properties determine metabolic needs.


Toward a more robust lower neck compressive injury tolerance-an approach combining multiple test methodologies.
The compressive tolerance of the cervical spine has traditionally been reported in terms of axial force at failure. Previous studies suggest that axial compressive force at failure is particularly sensitive to the alignment of the cervical vertebra and the end conditions of the test methodology used. The present study was designed to develop a methodology to combine the data of previous experiments into a diverse data set utilizing multiple test methods to allow for the evaluation of the robustness of current and proposed eccentricity based injury criteria. Data were combined from 2 studies composed of dynamic experiments including whole cervical spine and head kinematics that utilized different test methodologies with known end conditions, spinal posture, injury outcomes, and measured kinetics at the base of the neck. Loads were transformed to the center of the C7-T1 intervertebral disc and the eccentricity of the sagittal plane resultant force relative to the center of the disc was calculated. The correlation between sagittal plane resultant force and eccentricity at failure was evaluated and compared to the correlation between axial force and sagittal plane moment and axial force alone. Accounting for the eccentricity of the failure loads decreased the scatter in the failure data when compared to the linear combination of axial force and sagittal plane moment and axial force alone. A correlation between axial load and sagittal plane flexion moment at failure (R² = 0.44) was identified. The sagittal plane extension moment at failure did not have an identified correlation with the compressive failure load for the tests evaluated in this data set (R² = 0.001). The coefficients of determination for the linear combinations of sagittal plane resultant force with anterior and posterior eccentricity are 0.56 and 0.29, respectively. These correlations are an improvement compared to the combination of axial force and sagittal plane moment. Results using the outlined approach indicate that the combination of lower neck sagittal plane resultant force and the anterior-posterior eccentricity at which the load is applied generally correlate with the type of cervical damage identified. These results show promise at better defining the tolerance for compressive cervical fractures in male postmortem human subjects (PMHS) than axial force alone. The current analysis requires expansion to include more tolerance data so the robustness of the approach across various applied loading vectors and cervical postures can be evaluated.


Increased prevalence of methicillin-resistant Staphylococcus aureus nasal colonization in household contacts of children with community acquired disease.
To measure Methicillin-resistant Staphylococcus aureus (MRSA) nasal colonization prevalence in household contacts of children with current community associated (CA)-MRSA infections (study group) in comparison with a group of household contacts of children without suspected Staphylococcus aureus infection (a control group). This is a cross sectional study. Cultures of the anterior nares were taken. Relatedness of isolated strains was tested using pulse field gel electrophoresis (PFGE). The prevalence of MRSA colonization in the study group was significantly higher than in the control group (18/77 (23%) vs 3/77 (3.9%); p ≤ 0.001). The prevalence of SA colonization was 28/77 (36%) in the study group and 16/77 (21%) in the control group (p = 0.032). The prevalence of SA nasal colonization among patients was 6/24 (25%); one with methicillin-susceptible S. aureus (MSSA) and 5 with MRSA. In the study (patient) group, 14/24 (58%) families had at least one household member who was colonized with MRSA compared to 2/29 (6.9%) in the control group (p = 0.001). Of 69 total isolates tested by PFGE, 40 (58%) were related to USA300. Panton-Valetine leukocidin (PVL) genes were detected in 30/52 (58%) tested isolates. Among the families with ≥1 contact colonized with MRSA, similar PFGE profiles were found between the index patient and a contact in 10/14 families. Prevalence of asymptomatic nasal carriage of MRSA is higher among household contacts of patients with CA-MRSA disease than control group. Decolonizing such carriers may help prevent recurrent CA-MRSA infections.


In Silico Screening Identified Novel Small-molecule Antagonists of PAC1 Receptor.
Pituitary adenylate cyclase-activating polypeptide (PACAP) and its receptors are present in the spinal dorsal horn and dorsal root ganglia, suggesting an important role of PACAP signaling systems in the modulation of spinal nociceptive transmission. Previously, we found that intrathecal injection of PACAP or maxadilan, a selective PACAP type I (PAC1) receptor agonist, induced transient aversive responses followed by a long-lasting mechanical allodynia in mice, suggesting that PACAP-PAC1 receptor systems are involved in chronic pain and that selective PAC1 antagonists may become a new class of analgesics. Although several PAC1 antagonists, such as PACAP 6-38, have been reported, all of them are peptide compounds. In the present study, we identified new small-molecule antagonists of the PAC1 receptor using in silico screening and in vitro/vivo pharmacological assays. The identified small-molecule compounds, named PA-8 and PA-9, dose dependently inhibited the phosphorylation of CREB induced by PACAP in PAC1-, but not VPAC1- or VPAC2-receptor-expressing CHO cells. PA-8 and PA-9 also dose dependently inhibited PACAP-induced cAMP elevation with an IC50 of 2.0 and 5.6 nM, respectively. In vivo pharmacological assays showed that intrathecal injection of these compounds blocked the induction of PACAP-induced aversive responses and mechanical allodynia in mice. In contrast, the compounds when administered alone exerted neither agonistic nor algesic actions in the in vitro/vivo assays. The compounds identified in the present study are new and the first small-molecule antagonists of the PAC1 receptor; they may become seed compounds for developing novel analgesics.


Crackling versus continuumlike dynamics in brittle failure.
We study how the loading rate, specimen geometry, and microstructural texture select the dynamics of a crack moving through an heterogeneous elastic material in the quasistatic approximation. We find a transition, fully controlled by two dimensionless variables, between dynamics ruled by continuum fracture mechanics and crackling dynamics. Selection of the latter by the loading, microstructure, and specimen parameters is formulated in terms of scaling laws on the power spectrum of crack velocity. This analysis defines the experimental conditions required to observe crackling in fracture. Beyond failure problems, the results extend to a variety of situations described by models of the same universality class, e.g., the dynamics in wetting or of domain walls in amorphous ferromagnets.


Practical in situ-generation of phosphinite ligands for palladium-catalyzed carbonylation of (hetero)aryl bromides forming esters.
An effective method for alkoxycarbonylation of (hetero)aryl bromides is developed in the presence of in situ-generated phosphinite ligands tBu2POR (R = nBu, nPr, Et or Me). For this purpose commercially available tBu2PCl was used as the pre-ligand in the presence of different alcohols. For the first time cross coupling reactions with two alcohols - one generating the ligand, the other used as substrate - were developed. Through this method, ligand optimization can be performed in a more efficient manner and the desired products could be obtained with good yields and selectivity.


Frequency and impact of grade three or four toxicities of novel agents on outcomes of older patients with chronic lymphocytic leukemia and non-Hodgkin lymphoma (alliance A151611).
Older patients with cancer suffer from chemotherapy-related toxicities more frequently than younger patients. As novel agents are being used more commonly in chronic lymphocytic leukemia (CLL) and non-Hodgkin lymphoma (NHL), toxicities of these agents in older patients have not been well studied. Further, impact of these toxicities on outcomes in the elderly is unknown. This study aimed to answer both questions. We reviewed 14 Alliance for Clinical Trials in Oncology trials that enrolled CLL and/or NHL patients between 2004-2014. Toxicity was assessed per the NCI-CTCAE (version 3-5). Probabilities of experiencing grade three or four hematologic and non-hematologic toxicities were modeled as a function of clinical and disease-related factors using logistic regression. 1199 patients (409 age ≥ 65; 790 age < 65) were analyzed; 438 received only biologic therapy (145 age ≥ 65; 293 age < 65), and 761 received biologic + chemotherapy (264 age ≥ 65; 497 age < 65). The odds of grade three or four hematologic [odds ratio (OR) 1.70; p = 0.009: 95% CI (1.57-1.84)] and non-hematologic toxicities [OR 1.47; p = 0.022; 95% CI (1.39-1.55)] were increased in older patients with CLL, as well as odds of grade three or four non-hematologic toxicities [OR 1.89; p = 0.017; 95% CI (1.64-2.17)] in older patients with NHL. Grade three or four hematologic toxicities were associated with inferior OS and PFS in older patients with NHL [HR 3.14; p = 0.006; 95% CI (2.25-4.39) for OS and 3.06; p = 0.011; 95% CI (2.10-4.45) for PFS], though not in CLL. A prognostic model predicting grade three or four toxicities was also developed. CLL and NHL patients ≥ 65 year encounter more toxicities than younger patients even when treated with novel biologic agents. Development of grade three or four hematologic toxicities lead to inferior PFS and OS in NHL but not in CLL.


Web-based computer adaptive assessment of individual perceptions of job satisfaction for hospital workplace employees.
To develop a web-based computer adaptive testing (CAT) application for efficiently collecting data regarding workers' perceptions of job satisfaction, we examined whether a 37-item Job Content Questionnaire (JCQ-37) could evaluate the job satisfaction of individual employees as a single construct. The JCQ-37 makes data collection via CAT on the internet easy, viable and fast. A Rasch rating scale model was applied to analyze data from 300 randomly selected hospital employees who participated in job-satisfaction surveys in 2008 and 2009 via non-adaptive and computer-adaptive testing, respectively. Of the 37 items on the questionnaire, 24 items fit the model fairly well. Person-separation reliability for the 2008 surveys was 0.88. Measures from both years and item-8 job satisfaction for groups were successfully evaluated through item-by-item analyses by using t-test. Workers aged 26 - 35 felt that job satisfaction was significantly worse in 2009 than in 2008. A Web-CAT developed in the present paper was shown to be more efficient than traditional computer-based or pen-and-paper assessments at collecting data regarding workers' perceptions of job content.


Fermented food intake is associated with a reduced likelihood of atopic dermatitis in an adult population (Korean National Health and Nutrition Examination Survey 2012-2013).
The prevalence of atopic dermatitis (AD) has continuously increased throughout the world in every age group, and the recent increase in AD in Korean adults may be related to changes in nutrient intakes due to westernization of dietary patterns. We hypothesized that the prevalence of AD is associated with the different dietary patterns and fermented food intakes of the Korean adult population. We examined the hypothesis using 9763 adults 19 years or older using the 2012-2013 Korean National Health and Nutrition Examination Survey. We identified 4 dietary patterns in addition to that including fermented foods using principal component analysis on data obtained from a 116-item validated semiquantitative food frequency questionnaire: meat and processed foods; vegetables, fruits, legumes, seafood, and seaweed; rice and grains; and coffee, chocolate, and ice cream. Adjusted odds ratios (ORs) for AD were calculated according to dietary patterns after adjusting for potential confounders. High levels of consumption (>92 times/month) of fermented foods such as doenjang, chungkookjang, kimchi, fermented seafood, makgeolli, and beer were associated with a lower prevalence of AD (OR, 0.56; 95% confidence interval [CI], 0.37-0.84). In contrast, high levels of consumption of meat and processed foods were strongly associated with the prevalence of AD (OR, 2.42; 95% CI, 1.48-3.94). Interestingly, the consumption of coffee, chocolate, and ice cream was significantly negatively associated with the prevalence of AD (OR, 0.53; 95% CI, 0.34-0.82). In conclusion, the hypothesis was accepted. The results can be applied to nutrition education programs for the general population to decrease risk factors for AD.


Reoccurrence of botulinum neurotoxin subtype A3 inducing food-borne botulism, Slovakia, 2015.
A case of food-borne botulism occurred in Slovakia in 2015. Clostridium botulinum type A was isolated from three nearly empty commercial hummus tubes. The product, which was sold in Slovakia and the Czech Republic, was withdrawn from the market and a warning was issued immediately through the European Commission's Rapid Alert System for Food and Feed (RASFF). Further investigation revealed the presence of botulinum neurotoxin (BoNT) subtype BoNT/A3, a very rare subtype implicated in only one previous outbreak (Loch Maree in Scotland, 1922). It is the most divergent subtype of BoNT/A with 15.4% difference at the amino acid level compared with the prototype BoNT/A1. This makes it more prone to evading immunological and PCR-based detection. It is recommended that testing laboratories are advised that this subtype has been associated with food-borne botulism for the second time since the first outbreak almost 100 years ago, and to validate their immunological or PCR-based methods against this divergent subtype.


Vermicious thermo-responsive Pickering emulsifiers.
Thermo-responsive vermicious (or worm-like) diblock copolymer nanoparticles prepared directly in n-dodecane via polymerisation-induced self-assembly (PISA) were used to stabilise water-in-oil Pickering emulsions. Mean droplet diameters could be tuned from 8 to 117 μm by varying the worm copolymer concentration and the water volume fraction and very high worm adsorption efficiencies (∼100%) could be obtained below a certain critical copolymer concentration (∼0.50%). Heating a worm dispersion up to 150 °C led to a worm-to-sphere transition, which proved to be irreversible if conducted at sufficiently low copolymer concentration. This affords a rare opportunity to directly compare the Pickering emulsifier performance of chemically identical worms and spheres. It is found that the former nanoparticles are markedly more efficient, since worm-stabilised water droplets are always smaller than the equivalent sphere-stabilised droplets prepared under identical conditions. Moreover, the latter emulsions are appreciably flocculated, whereas the former emulsions proved to be stable. SAXS studies indicate that the mean thickness of the adsorbed worm layer surrounding the water droplets is comparable to that of the worm cross-section diameter determined for non-adsorbed worms dispersed in the continuous phase. Thus the adsorbed worms form a monolayer shell around the water droplets, rather than ill-defined multilayers. Under certain conditions, demulsification occurs on heating as a result of a partial worm-to-sphere morphological transition.


IL-1 does not reverse the anti-proliferative effect of aspirin in breast cancer cells.
Interleukin-1 (IL-1) is a multifunctional proinflammatory cytokine. There have been studies suggesting a role in affecting growth and invasiveness of malignant breast cells by either blocking or stimulating growth of cultured MCF-7 breast cancer cells. This effect may be mediated by induction of COX-2. Aspirin is an inhibitor of COX-2 and has been implicated, with other non-steroidal anti-inflammatory drugs (NSAIDS) in prevention and treatment of breast cancer. In this study the in vitro effects of IL-1 and aspirin on growth of MCF-7 human breast cancer cells was examined. MCF-7 cells were treated with various concentrations of IL-1 and aspirin alone and in combination. Cell growth was assessed by cell number measurement. Aspirin significantly decreased growth rate in a dose-dependent manner, alone and as a combined treatment with IL-1 with a maximum reduction in growth rate at 300 mg/ml (P < 0.05). Treatment with IL-1 alone showed no significant effect on growth rate of MCF-7 cells (P > 0.05). This study confirms that aspirin suppresses the proliferation rate of MCF-7 cells both as a single agent and in combination with IL-1. It also suggests that IL-1 alone does not stimulate or inhibit growth of MCF-7 cells.


A simple two-component model of the electrically evoked compound action potential in the human cochlea.
Neural response telemetry (NRT) permits in situ intracochlear recordings of the electrically evoked compound action potential from the auditory nerves using scala tympani electrodes. The recorded NRT waveforms can generally be categorized under either single positive peak or double positive peak waveforms. This is similar to the observations from Stypulkowski and van den Honert, who suggested that the double peak complex arises from two components that could be axonal and dendritic in origin, respectively. Using a simple mathematical model which linearly combines two separate waveforms similar in shape but differing in amplitude and latencies, it was possible to simulate the various NRT waveform categories. The simulation results support the view that the two waveform components originate from dendritic or axonal processes and implies that the shape of the response waveform may provide information about the degree of neural survival in the stimulated cochlea. This information could be useful for determining optimal speech coding parameters for cochlear implant users on an individual basis.


Cirrus high-definition optical coherence tomography compared with Stratus optical coherence tomography in glaucoma diagnosis.
To compare the retinal nerve fiber layer (RNFL) evaluation using Cirrus optical coherence tomography (OCT) and Stratus OCT in glaucoma diagnosis. One hundred thirty normal and 86 patients with glaucoma were included in this prospective study. The signal strengths of the OCTs were evaluated. The sensitivities and specificities of global RNFL average thickness were compared in the four quadrants and in each clock hour sector. Receiver operating characteristic (ROC) curves, areas under the ROC (AUC), and the likelihood ratio (LR) were plotted for RNFL thickness. Agreement between the OCTs was calculated by using the Bland-Altman method and kappa (kappa) coefficient. Twenty-three percent of all cases examined with Stratus OCT and 1.9% examined with Cirrus OCT had a signal strength below 6 (P = 0.01). In cases with signal strengths > or =6, the mean signal strength was higher with Cirrus OCT than with Stratus OCT (P = 0.01). The RNFL measurements by Cirrus were thicker than those of Stratus OCT (P < 0.05). The AUCs were 0.829 for Stratus and 0.837 for Cirrus OCT (P = 0.706) for global RNFL average. LRs were similar in both OCTs in global RNFL classification but varied in quadrants. The widths of the limits of agreement varied between 42.16 and 97.79 microm. There was almost perfect agreement (kappa = 0.82) in the average RNFL classification. Cirrus OCT has better scan quality than Stratus OCT, especially in glaucomatous eyes. In cases with good-quality scans, the sensitivity and specificity, and AUCs were similar. The best agreement was in the global average RNFL classification. The widths of limits of agreements exceed the limits of resolution of the OCTs.


Disparities in scientific research activity between doctors and nurses working in the Peruvian health care system: Analysis of a nationally representative sample.
To evaluate disparities in the frequency of scientific activity between medical doctors and nurses in Peru. We carried out a secondary data analysis of the National Health Services Users' Satisfaction Survey (ENSUSALUD), 2016. This nationally representative survey evaluates doctors and nurses working in clinical settings. We defined scientific activity as i) having published an original article (journal indexed in Web of Science, Scopus or Medline); and ii) having authored an abstract in a national or international conference. We estimated crude and adjusted disparities prevalence ratios (aDPR) and 95% confidence intervals (95%CI). We included 2025 doctors and 2877 nurses in the analysis; 71% of doctors doctor were male, and 93% of nurses were female (p<0.001). Among doctors, 13.9% had published an article, and 8.4% presented an abstract at a conference in the last two years, while these proportions were 0.6% and 2.5% for nurses, respectively. The adjusted models showed that doctors, when compared to nurses, were approximately 27 times likely to have published a paper (aDPR = 27.86; 95% CI 10.46 to 74.19) and twice as likely to have authored a conference abstract (aDPR = 2.51; 95% CI 1.39 to 4.53). There are important disparities in scientific activity between doctors and nurses working in clinical settings in Peru. Disparities are more significant for article publication than for authoring in conference abstracts. We suggest public policies that promote research dissemination between health professionals, with emphasis on nurses.


4D multi-modality tissue segmentation of serial infant images.
Accurate and consistent segmentation of infant brain MR images plays an important role in quantifying patterns of early brain development, especially in longitudinal studies. However, due to rapid maturation and myelination of brain tissues in the first year of life, the intensity contrast of gray and white matter undergoes dramatic changes. In fact, the contrast inverse around 6-8 months of age, when the white and gray matter tissues are isointense and hence exhibit the lowest contrast, posing significant challenges for segmentation algorithms. In this paper, we propose a longitudinally guided level set method to segment serial infant brain MR images acquired from 2 weeks up to 1.5 years of age, including the isointense images. At each single-time-point, the proposed method makes optimal use of T1, T2 and the diffusion-weighted images for complimentary tissue distribution information to address the difficulty caused by the low contrast. Moreover, longitudinally consistent term, which constrains the distance across the serial images within a biologically reasonable range, is employed to obtain temporally consistent segmentation results. Application of our method on 28 longitudinal infant subjects, each with 5 longitudinal scans, shows that the automated segmentations from the proposed method match the manual ground-truth with much higher Dice Ratios than other single-modality, single-time-point based methods and the longitudinal but voxel-wise based methods. The software of the proposed method is publicly available in NITRC (http://www.nitrc.org/projects/ibeat).


RIBBON-2: a randomized, double-blind, placebo-controlled, phase III trial evaluating the efficacy and safety of bevacizumab in combination with chemotherapy for second-line treatment of human epidermal growth factor receptor 2-negative metastatic breast cancer.
This phase III study compared the efficacy and safety of bevacizumab combined with standard chemotherapy regimens versus chemotherapy alone as second-line treatment of patients with human epidermal growth factor receptor 2 (HER2) -negative metastatic breast cancer. Patients were randomly assigned 2:1 to chemotherapy + bevacizumab or to chemotherapy + placebo. Before random assignment, investigators chose capecitabine, a taxane (paclitaxel, nab-paclitaxel, or docetaxel), gemcitabine, or vinorelbine. Dosing for bevacizumab or placebo was 15 mg/kg every 3 weeks or 10 mg/kg every 2 weeks, depending on chemotherapy regimen. The primary end point was progression-free survival (PFS). Secondary end points included overall survival, PFS by chemotherapy cohort, objective response rate (ORR), duration of objective response, 1-year survival rate, and safety. RIBBON-2 enrolled 684 patients (225, chemotherapy + placebo; 459, chemotherapy + bevacizumab). The combination of bevacizumab with chemotherapy demonstrated a statistically significant benefit. Median PFS increased from 5.1 to 7.2 months (stratified hazard ratio for PFS, 0.78; 95% CI, 0.64 to 0.93; P = .0072). The 10% improvement in ORR between the placebo- and bevacizumab-containing arms (39.5% v 29.6%; P = .0193), although not statistically significant, was consistent with previous trials. There was no statistically significant difference in overall survival. The most common grade ≥ 3 adverse events (AEs) related to bevacizumab treatment were hypertension (9.0%) and proteinuria (3.1%). There was an increased number of AEs leading to study discontinuation in the chemotherapy + bevacizumab arm compared with the chemotherapy + placebo arm (13.3% v 7.2%). The combination of bevacizumab with commonly used chemotherapies improved PFS in the second-line treatment of patients with HER2-negative metastatic breast cancer, with a safety profile comparable with that in prior phase III studies.


Detection of extracapsular extension of prostate cancer: role of fat suppression endorectal MRI.
The purpose of this work was to compare the efficacy of fat-suppressed and non-fat-suppressed fast spin echo (FSE) endorectal MRI in the detection of extracapsular extension (ECE) of prostate cancer by experienced and inexperienced readers. Seventy-nine patients with biopsy-proven prostate cancer underwent axial FSE T2-weighted endorectal MRI of the prostate prior to radical prostatectomy. Twenty-one patients were imaged with frequency-selective fat suppression, and 58 were imaged without fat suppression. All images were retrospectively and independently reviewed by two readers of different experience levels who were blinded to clinical and pathological findings. Readers assessed the presence or absence of ECE on a 5 point scale for each side of the prostate, and step-section pathology was used as the standard of reference in all patients. Receiver operating characteristics analysis was used to compare the performance of fat-suppressed and non-fat-suppressed images by both readers. ECE was present in 33 of 79 (42%) patients. The more experienced reader demonstrated better diagnostic performance (p < 0.05) than the less experienced reader in terms of sensitivity and area under the ROC curve (Az) for MRI without fat suppression. Use of frequency-selective fat suppression did not result in any significant improvement in diagnosis of ECE compared with MRI without fat suppression for either the experienced (Az 0.81 vs. 0.79) or the inexperienced (Az 0.76 vs. 0.68) reader. Even when reader experience is considered, use of frequency-selective fat suppression did not significantly improve the diagnosis of ECE by MRI. The decision to use fat suppression and the selection of a fat suppression technique can be left to the discretion of the individual reader.


Stereoisomeric profiling of chiral pharmaceutically active compounds in wastewaters and the receiving environment - A catchment-scale and a laboratory study.
Chiral pharmaceutically active compounds (cPACs) are not currently governed by environmental regulation yet are expected to be in the future. As cPACs can exert stereospecific toxicity in the aquatic environment, it is essential to better understand their stereoselective behaviour here. Therefore, this study aims to provide a new perspective towards comprehensive evaluation of cPACs at a river catchment level, including their stereochemistry as a chemical phenomenon driving fate of chiral molecules in the environment. A large spatial and temporal monitoring program was performed in Southwest England. It included 5 sewage treatment works and the receiving waters of the largest river catchment in Southwest England. Simultaneously, lab-scale microcosm studies in simulated activated sludge bioreactors and river water microcosm were performed to evaluate stereoselective degradation of cPACs. A multi-residue enantioselective method allowed the analysis of a total of 18 pairs of enantiomers and 3 single enantiomers in wastewater and river water samples. Our monitoring program revealed: (1) spatial and temporal variations of cPACs in influent wastewaters resulting from different patterns of usage as well as an (2) enantiomeric enrichment of cPACs, likely due to human metabolism, despite their commercialization as racemic mixtures. A similar chiral signature was observed in effluent and receiving waters. Stereoselective degradation was observed in trickling filters (TF) for naproxen, ketoprofen, cetirizine and 10,11-dihydroxy-10-hydroxycarbamazepine, in sequencing batch reactors (SBR) for ifosfamide and in activated sludge (AS) for cetirizine. The extent of enantiomer-specific fate was wastewater treatment dependent in the case of naproxen (TF showed higher stereoselectivity than AS and SBR) and cetirizine (TF and AS showed higher stereoselectivity than SBR) due to differing microbial population. Furthermore, stereoselective degradation of naproxen was highly variable among STWs using similar treatments (TF) and operating in the same region. Microbial stereoselective degradation was also confirmed by both activated and river water simulated microcosm for chloramphenicol, ketoprofen, indoprofen, naproxen and 10,11-dihydroxy-10-hydroxycarbamazepine. Results from our large scale river catchment monitoring study and lab simulated microcosm show wide-ranging implications of enantiomerism of cPACs on environmental risk assessment (ERA). As two enantiomers of the same compound show different biological effects (e.g. toxicity), their non-racemic presence in the environment might lead to inaccurate ERA. This is because current ERA approaches do not require analysis at enantiomeric level.


The Heidelberg Screen for Pattern Mutants of Drosophila: A Personal Account.
In large-scale mutagenesis screens performed in 1979-1980 at the EMBL in Heidelberg, we isolated mutations affecting the pattern or structure of the larval cuticle in Drosophila. The 600 mutants we characterized could be assigned to 120 genes and represent the majority of such genes in the genome. These mutants subsequently provided a rich resource for understanding many fundamental developmental processes, such as the transcriptional hierarchies controlling segmentation, the establishment of cell states by signaling pathways, and the differentiation of epithelial cells. Most of the Heidelberg genes are now molecularly known, and many of them are conserved in other animals, including humans. Although the screens were initially driven entirely by curiosity, the mutants now serve as models for many human diseases. In this review, we describe the rationale of the screening procedures and provide a classification of the genes on the basis of their initial phenotypes and the subsequent molecular analyses.


Heparin-induced antibodies and cardiovascular risk in patients on dialysis.
The clinical relevance of heparin-induced antibodies (HIA) in the absence of thrombocytopenia remains to be defined. The aims of this study were (i) to determine the prevalence of HIA in patients treated by dialysis, (ii) to determine the prevalence of thrombocytopenia and heparin-induced thrombocytopenia (HIT), and (iii) to test whether HIA are associated with adverse outcomes. Sera from 740 patients treated by hemodialysis (HD, n=596) and peritoneal dialysis (PD, n=144) were tested for HIA (IgG, IgA or IgM) by masked investigators at approximately six months after enrolment in the Choices for Healthy Outcomes in Caring for End-Stage Renal Disease (CHOICE) study. We assessed, with time-to-event Cox proportional hazards models, whether the presence of HIA predicted any of four clinical outcomes: arterial cardiovascular events, venous thromboembolism, vascular access occlusion and mortality. HIA prevalence was 10.3% overall. HIA positivity did not predict development of thrombocytopenia or any of the four clinical outcomes over a mean follow-up of 3.6 years, with hazard ratios for arterial cardiovascular events of 0.98 (95% confidence interval 0.70-1.37), venous thromboembolism 1.39 (0.17-11.5), vascular access occlusion 0.82 (0.40-1.71), and mortality 1.18 (0.85-1.64). Chronic intermittent heparin exposure was associated with a high seroprevalence of HIA. In dialysis patients these antibodies were not an independent risk factor for cardiovascular events and mortality. Our data do not suggest that dialysis patients should be monitored for HIA antibodies in the absence of thrombocytopenia.


Transcutaneous immunization with novel lipid-based adjuvants induces protection against gastric Helicobacter pylori infection.
The development of vaccines to combat pathogens that infect across mucosal surfaces has been a major goal of vaccine research. Successful mucosal vaccination requires the co-administration of adjuvants that can overcome the state of immune tolerance normally associated with mucosal application of proteins. In the case of oral immunization, delivery systems are also required to protect vaccine antigens against destruction by gastric pH and digestive enzymes. Furthermore, adjuvants used for mucosal delivery must be free of neurotoxic effects like those induced by the commonly used experimental mucosal adjuvant cholera toxin. Maintenance of the "cold chain" is also essential for the effectiveness of any vaccine and adjuvants/delivery systems that enhance the stability of a vaccine would offer a significant advantage. Needle-free methods of vaccination that induce protective immunity at multiple mucosal surfaces are also desirable for rapid vaccination of large populations. In the present study we show that transcutaneous immunization (TCI) using Lipid C, a novel lipid-based matrix originally developed for oral immunization, containing soluble Helicobacter sonicate significantly reduces the gastric bacterial burden in mice following gastric challenge with live Helicobacter pylori. Protection is associated with the production of splenic gamma interferon and gastric IgA and was achieved without the co-administration of potent and potentially toxic adjuvants, although protection was further enhanced by inclusion of CpG-ODN and cholera toxin in the lipid delivery system.


Car drivers attend to different gaze targets when negotiating closed vs. open bends.
On winding roads, car drivers have to control speed and steering angle in order to keep the car in an optimal lane position. Among the strategies proposed for steering regulation are the use of the tangent point, a geometrical method, and gaze sampling, in which retinal flow lines obtained by tracking a spot on the future road need to be assessed. Previous studies used a variety of scenarios (real-road vs. simulator) and different road designs (closed vs. open bends, different curvatures) and found results speaking in favor of either strategy. Here, we investigate what effects the openness of the bend, i.e. the sight distance of the driver, has on the percentage with which drivers use the tangent point. Six drivers drove a test car repeatedly through a series of twelve bends on real roads while their eye-movements were recorded. Results show that the reliance on the tangent point is generally high and increases with the closedness (shorter sight distances) of the bend and higher curvature. In open bends they alternatively look far into the straight road segments adjacent to the bend, but do not use gaze sampling.


Pilot study assessing balance in women with fibromyalgia syndrome.
The purpose of current study was to assess postural control and balance self-efficacy in people with fibro-myalgia syndrome [FMS]. Thirty-two females with FMS completed the Activity-specific Balance Confidence Scale, Berg Balance test, NeuroCom Balance Master sensory-organization test, and limits of stability. There was a high prevalence of reported falls and a low mean score on the Activity-specific Balance Confidence Scale. A significant number of subjects scored below the population norm fifth percentile score on the sensory-organization test composite, visual, and vestibular sections. Preliminary evidence suggests that women with FMS may present with deficits in postural control, sensory organization, and balance self-efficacy.


PAEDIATRIC STANDARDISED CONCENTRATION INFUSIONS - A NATIONAL SOLUTION.
Wide scale implementation of paediatric standardised concentration infusions (SCIs) and the use of smart pump technology has been slow despite international safety agency recommendations. Implementation rates in European hospitals fall far below those in the United States, where for the last decade accreditation has been linked to implementation.1 2 Multidisciplinary collaboration is essential, with pharmacy input and the creation of a smart pump drug library recognised as often being limiting, yet crucial factors, to implementation.3 Following on from the successful development and implementation of a paediatric drug library of standardised concentrations in the paediatric intensive care unit (PICU), operating theatres and cardiac ward of a large tertiary children's hospital, a project was set up to further develop this library for use across multiple sites including paediatric acute transport services. Post-implementation of the original SCI drug library in the single site paediatric hospital in 2012, a database was created to record proposals for amendments and expansion of that library. A cross-site multidisciplinary collaborative working group was established with representation from PICU pharmacists, intensivists, nursing and clinical engineering to progress multisite adoption of a standardised drug library. Differences in practices across sites were identified and resolved by consensus where possible. Unresolved differences were overcome by omission or 'hiding' of particular drug lines at individual sites until consensus could be reached for future updates. Legally binding agreements were drawn up between sites, in conjunction with the infusion pump vendors, to prevent any future deviations from the master library. Cross-site collaboration over a number of months facilitated the successful amendment and extension of the original drug library to create a new master drug file. Individual site versions of this library, and the supporting documentation were created and disseminated. A change control management plan was developed and agreed upon. Cross-site collaboration is achievable in supporting the increased implementation of standardised concentration infusions. The standardisation of practices across sites maximises both human and financial resources, and has the potential to reduce medications errors as both patients and medical staff transfer across sites.