Metabolism of butoxyethanol in excised human skin in vitro.
Glycol ethers are widely used in industrial and household applications because their chemical and physical properties make them versatile solvents, miscible with both water and organic media. Due to the ease with which the glycol ethers are absorbed through the skin and the potential for development of adverse health effects it is important to understand the extent to which local metabolism can contribute to local and systemic toxicity. Sections of previously frozen, full thickness excised human skin samples were placed on transwell supports and placed with the underside of the skin in contact with receptor fluid. The skin surface was dosed with 115.2 mg of neat butoxyethanol and the absorption and metabolism of butoxyethanol to butoxyacetic acid monitored over time. In total 64.94+/-0.04 mg of butoxyethanol or its metabolites were removed from the surface of the skin at 24h, representing the equivalent of 56% of the applied dose, the equivalent of 17.5% of the applied dose was recovered from the receiver fluid, 3% from within the skin and the remaining 23.5% of the dose was lost to the atmosphere through evaporation. After 24h a total of 31.5 microg of butoxyacetic acid had been produced representing approximately 0.03% of the applied dose. Therefore approximately 0.16% (31.5 microg as a percentage of the total amount of butoxyethanol reaching the receiver fluid (20.17 mg) of the absorbed butoxyethanol was metabolised to butoxyacetic acid during its passage through the skin. This suggested that, although enzyme activities capable of converting butoxyethanol to butoxyacetic acid are present in skin, metabolic conversion during percutaneous absorption was small and systemic exposure to the parent compound rather than the metabolite would occur following dermal exposure to butoxyethanol. This experiment demonstrates that it is possible to maintain metabolic activity in skin samples in an in vitro setup for short, but experimentally useful, periods.


Structure and protein-protein interaction studies on Chlamydia trachomatis protein CT670 (YscO Homolog).
Comparative genomic studies have identified many proteins that are found only in various Chlamydiae species and exhibit no significant sequence similarity to any protein in organisms that do not belong to this group. The CT670 protein of Chlamydia trachomatis is one of the proteins whose genes are in one of the type III secretion gene clusters but whose cellular functions are not known. CT670 shares several characteristics with the YscO protein of Yersinia pestis, including the neighboring genes, size, charge, and secondary structure, but the structures and/or functions of these proteins remain to be determined. Although a BLAST search with CT670 did not identify YscO as a related protein, our analysis indicated that these two proteins exhibit significant sequence similarity. In this paper, we report that the CT670 crystal, solved at a resolution of 2 A, consists of a single coiled coil containing just two long helices. Gel filtration and analytical ultracentrifugation studies showed that in solution CT670 exists in both monomeric and dimeric forms and that the monomer predominates at lower protein concentrations. We examined the interaction of CT670 with many type III secretion system-related proteins (viz., CT091, CT665, CT666, CT667, CT668, CT669, CT671, CT672, and CT673) by performing bacterial two-hybrid assays. In these experiments, CT670 was found to interact only with the CT671 protein (YscP homolog), whose gene is immediately downstream of ct670. A specific interaction between CT670 and CT671 was also observed when affinity chromatography pull-down experiments were performed. These results suggest that CT670 and CT671 are putative homologs of the YcoO and YscP proteins, respectively, and that they likely form a chaperone-effector pair.


Metabolic phenotypes, vascular complications, and premature deaths in a population of 4,197 patients with type 1 diabetes.
Poor glycemic control, elevated triglycerides, and albuminuria are associated with vascular complications in diabetes. However, few studies have investigated combined associations between metabolic markers, diabetic kidney disease, retinopathy, hypertension, obesity, and mortality. Here, the goal was to reveal previously undetected association patterns between clinical diagnoses and biochemistry in the FinnDiane dataset. At baseline, clinical records, serum, and 24-h urine samples of 2,173 men and 2,024 women with type 1 diabetes were collected. The data were analyzed by the self-organizing map, which is an unsupervised pattern recognition algorithm that produces a two-dimensional layout of the patients based on their multivariate biochemical profiles. At follow-up, the results were compared against all-cause mortality during 6.5 years (295 deaths). The highest mortality was associated with advanced kidney disease. Other risk factors included 1) a profile of insulin resistance, abdominal obesity, high cholesterol, triglycerides, and low HDL(2) cholesterol, and 2) high adiponectin and high LDL cholesterol for older patients. The highest population-adjusted risk of death was 10.1-fold (95% CI 7.3-13.1) for men and 10.7-fold (7.9-13.7) for women. Nonsignificant risk was observed for a profile with good glycemic control and high HDL(2) cholesterol and for a low cholesterol profile with a short diabetes duration. The self-organizing map analysis enabled detailed risk estimates, described the associations between known risk factors and complications, and uncovered statistical patterns difficult to detect by classical methods. The results also suggest that diabetes per se, without an adverse metabolic phenotype, does not contribute to increased mortality.


Mammographic density and matrix metalloproteinases in breast tissue.
Mammographic density is a strong risk factor for breast cancer, yet the underlying histopathologic correlates are not clear. Matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) play important roles in multiple stages of tumorigenesis. This study examined the association between mammographic density and expression of MMPs 1, 3, 9, and 12 and TIMP3 in benign and malignant breast tissue of 277 women with mainly Caucasian and Japanese ancestry. Tissue microarrays with up to 4 benign and 4 malignant cores per woman were stained immunohistochemically and evaluated. Digitized prediagnostic mammograms were assessed for densities using a computer-assisted method. General linear models adjusted for known confounders were applied to estimate mean densities by staining category. Strong expression of all MMPs was about twice as frequent in malignant as in benign tissue, while TIMP3 expression in stromal tissue was higher in benign than malignant cores. For MMP3 and 9, less than 10% of cores stained positive; thus, they were not further analyzed. None of the markers showed a statistically significant association with breast density in the entire study population and ethnic-specific results were conflicting and difficult to explain. Although not statistically significant, mean density was consistently lower with more extensive TIMP3 expression in stromal and epithelial tissue. These findings indicate that the higher breast cancer risk in women with dense breasts may be influenced by lower TIMP3 expression. However, future investigations into activities and ratios of additional proteases and their inhibitors as well as other pathways, such as inflammation, are needed.


Sociodemographics, lifestyle factors and health status indicators associated with alcohol consumption and related behaviours: a Brazilian population-based analysis.
To investigate how various alcohol-drinking behaviours are associated with sociodemographics, lifestyle factors and health status indicators in Brazil. This study is based on a household survey of 53,034 adults aged 18 + years from all 26 Brazilian capitals and the Federal District conducted in 2017. Sex-stratified relationships were modelled using logistic regressions and controlled for capital-specific effects. Main outcome measures included regular alcohol use, weekly alcohol use, heavy episodic drinking (HED), frequent HED and drinking and driving. Overall (unadjusted) prevalence of regular alcohol consumption is 41%. Among drinkers, approximately 70% drink on a weekly basis, and 46% are heavy episodic drinkers. Among this latter group, close to 44% are frequent heavy episodic drinkers (i.e. at least four times in a month). Among regular drinkers who also are drivers, the prevalence of drinking and driving is 28%. These prevalences are considerably higher in men. The relationships investigated vary by drinking behaviour and sex, with some factors consistently associated with various behaviours, when present. Population (men or women) at greatest risk include (largely) younger individuals (up to 700% increase in odds) who are single or divorced, those who are less health conscious and watch television or use mobile devices during leisure time 4 + hours per day and do not have diabetes. For drinking and driving, the additional risk factors include speeding behaviour, the use of mobile devices while driving and HED. Education, race/ethnicity and other health status indicators are differently associated with various drinking behaviours. For women, in particular, the results also show differences in odds of up to 360% and 1430% across cities for frequent HED and drinking and driving, respectively. Similarly, indigenous women are at greatest risk of weekly alcohol use and HED. HED and drinking and driving are problematic, as the association with other factors suggests a clustering of risky behaviours that may exacerbate the consequences of drinking behaviours.


Pharyngeal airway dimensions: a cephalometric, growth-study-based analysis of physiological variations in children aged 6-17.
The aim was to assess pharyngeal airway dimensions and physiological changes based on lateral cephalometric radiographs from healthy untreated children aged 6-17 years. The sample consisted of 880 lateral cephalograms (412 females and 468 males) of the Zurich Craniofacial Growth Study. Statistical analyses on cephalometric measurements of airway dimensions (distances 'p': shortest distance between soft palate and posterior pharyngeal wall and 't': shortest distance between tongue and posterior pharyngeal wall) and craniofacial parameters were performed. To disclose differences between different age groups, a Kruskal-Wallis test was applied. The influence of gender on 'p' and 't' was analysed by a Mann-Whitney U-test for each age group separately. The Spearman correlation was computed in order to investigate associations between craniofacial parameters. Variables associated with 'p' and 't' were chosen for multiple regression model investigation. The results demonstrated high interindividual variations. A slight influence of age on 'p' (P = 0.034) could be attested (+1.03 mm) but not on 't' (P = 0.208). With the exception of the 9-year age group, no significant differences between the genders were found. Correlation analysis revealed several statistically significant correlations between 't' or 'p' and antero-posterior cephalometric variables. All correlation coefficients were, however, very low and the adjusted coefficient of determination also revealed the regression model to be very weak. The high interindividual variations of 'p' and 't' render the use of reference values problematic. Contrary to other craniofacial structures, neither age-related changes nor sexual dimorphism were found for 'p' and 't'. Any associations to antero-posterior cephalometric characteristics seem low.


Endarterectomy vs stenting for carotid artery stenosis: a systematic review and meta-analysis.
The relative efficacy and safety of endarterectomy and stenting in patients with carotid stenosis remain unclear. In this review we synthesize the available evidence derived from randomized controlled trials (RCTs) that compared the two procedures in terms of the risks of death, stroke (disabling and nondisabling), and nonfatal myocardial infarction. We searched for RCTs in MEDLINE, EMBASE, Current Contents, and Cochrane CENTRAL; expert files, and bibliographies of included articles. Two reviewers, working independently, determined trial eligibility and extracted descriptive, methodologic, and outcome data from each eligible RCT. Random-effects meta-analysis was used to assess relative and absolute risks and the I(2) statistic was used to assess heterogeneity of treatment effect among trials. Ten RCTs with 3182 participants proved eligible. At 30 days and compared with endarterectomy, carotid stenting was associated with a nonsignificant reduction in the risk of death (relative risk [RR], 0.61; 95% confidence interval [CI], 0.27-1.37; I(2) = 0%), a nonsignificant reduction in the risk of nonfatal myocardial infarction (RR, 0.43; 95% CI 0.17-1.11; I(2) = 0%), and a nonsignificant increase in the risk of any stroke (RR, 1.29; 95% CI, 0.73-2.26; I(2) = 40%) and major/disabling stroke (RR, 1.06; 95% CI, 0.32-3.52; I(2) = 45%). If one considers the two procedures equivalent if the absolute difference in events is <2%, these results provide moderate-quality evidence for equivalence with respect to death (risk difference [RD] -0.40, 95% CI -1.02 to 0.40) and nonfatal myocardial infarction (RD, -0.70; 95% CI -1.90 to 0.50), but because of much wider CI, only low-quality evidence of equivalence in stroke (RD, 1.00; 95% CI, -1.00 to 3.10). In RCTs, carotid stenting and carotid endarterectomy seem equivalent in terms of death and nonfatal myocardial infarction. Although the impact on stroke remains unestablished, results are consistent with a clinically important increase in stroke risk with stenting, an intervention that aims at reducing the risk of stroke.


Overexpression of eIF5 or its protein mimic 5MP perturbs eIF2 function and induces ATF4 translation through delayed re-initiation.
ATF4 is a pro-oncogenic transcription factor whose translation is activated by eIF2 phosphorylation through delayed re-initiation involving two uORFs in the mRNA leader. However, in yeast, the effect of eIF2 phosphorylation can be mimicked by eIF5 overexpression, which turns eIF5 into translational inhibitor, thereby promoting translation of GCN4, the yeast ATF4 equivalent. Furthermore, regulatory protein termed eIF5-mimic protein (5MP) can bind eIF2 and inhibit general translation. Here, we show that 5MP1 overexpression in human cells leads to strong formation of 5MP1:eIF2 complex, nearly comparable to that of eIF5:eIF2 complex produced by eIF5 overexpression. Overexpression of eIF5, 5MP1 and 5MP2, the second human paralog, promotes ATF4 expression in certain types of human cells including fibrosarcoma. 5MP overexpression also induces ATF4 expression in Drosophila The knockdown of 5MP1 in fibrosarcoma attenuates ATF4 expression and its tumor formation on nude mice. Since 5MP2 is overproduced in salivary mucoepidermoid carcinoma, we propose that overexpression of eIF5 and 5MP induces translation of ATF4 and potentially other genes with uORFs in their mRNA leaders through delayed re-initiation, thereby enhancing the survival of normal and cancer cells under stress conditions.


Spontaneously resolved uterine prolapse in a neonate with spina bifida.
Yapıcıoğlu Yıldızdaş H, Ece Ü, Bilen Yurdakul G, Kolağasıgil E. Spontaneously resolved uterine prolapse in a neonate with spina bifida. Turk J Pediatr 2019; 61: 979-981. Neonatal uterine prolapse is rare and mostly due to spina bifida. It is probably due to abnormal innervation of levator ani and subsequent atrophy of the muscle in patients with spina bifida. Here we report a newborn patient with uterine prolapse after meningomyelocele operation. Foley catheter was used to manage uterine prolapse, however rectal prolapse occurred and catheter was pulled out. She was followed in an outpatient clinic without any management. She is now seven months old and had no uterine prolapse since the age of two months.


The effects of two Chinese herbal medicinal formulae vs. placebo controls for treatment of allergic rhinitis: a randomised controlled trial.
Allergic rhinitis is a chronic illness, affecting 10 to 40% of the worldwide population. Chinese herbal medicines, the treatment of allergic rhinitis, adopted thousands of years in ancient China, has recently raised much attention among researchers globally. This study evaluates the effects of two Chinese herbal formulae [Cure-allergic-rhinitis Syrup (CS) and Yu-ping-feng San (YS)] in treating undergraduate nursing students with allergic rhinitis over a 3-month follow-up, when compared to a placebo control group. A double-blind, randomised controlled trial with repeated-measures, three-parallel-groups design was conducted in a random sample of 249 participants recruited from one university in Hong Kong. After baseline measurements, participants were randomly assigned to CS, YS, or placebo groups (n=83 per group). The main outcomes, including symptom severity, quality of life, and body constitution, were measured with self-administered questionnaires at baseline and immediately, 1 and 3 months after the 4-week interventions. 240 participants completed the trial, with 9 (3.6%) drop-outs. The results of Generalised Estimating Equations test followed by pairwise contrasts tests indicated that the participants who received CS showed significantly greater reduction of symptoms (mean difference of CS vs. placebo=26.13-34.55, P<0.0005) and improvements in quality of life (mean difference of CS vs. placebo=12.81-16.76, P<0.001), and body constitution in 'Qi-deficiency', 'Yang-deficiency', and 'Inherited Special' (mean difference of CS vs. placebo=7.05-8.12, 7.56-8.92, and 4.48-8.10, P=0.01-< 0.0005, 0.001-0.004, and 0.01-< 0.0005, accordingly, at three post-tests). The participants who received YS also indicated significant greater improvements in symptom severity, quality of life, and a few patterns of body constitution when compared to the placebo group. However, its effects were lesser in strength (i.e., smaller effect sizes), varieties of symptoms, and body constitution and sustainability over the 3 months. The herbal formula CS was found effective to reduce symptoms and enhance quality of life in young adults (nursing students) with allergic rhinitis in 'Yang- and/or Qi-deficiency' body constitution. Further controlled trials of its effects in Chinese and/or Asians with allergic rhinitis in terms of socio-demographic, ethnic and illness characteristics and a longer-term follow-up are recommended. The trial has registered at ClinicalTrials.gov with an ID: NCT02027194 (3 January 2014).


Investigating disease severity in an animal model of concurrent babesiosis and Lyme disease.
The incidence of babesiosis, Lyme disease and other tick-borne diseases has increased steadily in Europe and North America during the last five decades. Babesia microti is transmitted by species of Ixodes, the same ticks that transmit the Lyme disease-causing spirochete, Borrelia burgdorferi. B. microti can also be transmitted through transfusion of blood products and is the most common transfusion-transmitted infection in the U.S.A. Ixodes ticks are commonly infected with both B. microti and B. burgdorferi, and are competent vectors for transmitting them together into hosts. Few studies have examined the effects of coinfections on humans and they had somewhat contradictory results. One study linked coinfection with B. microti to a greater number of symptoms of overall disease in patients, while another report indicated that B. burgdorferi infection either did not affect babesiosis symptoms or decreased its severity. Mouse models of infection that manifest pathological effects similar to those observed in human babesiosis and Lyme disease offer a unique opportunity to thoroughly investigate the effects of coinfection on the host. Lyme disease has been studied using the susceptible C3H mouse infection model, which can also be used to examine B. microti infection to understand pathological mechanisms of human diseases, both during a single infection and during coinfections. We observed that high B. microti parasitaemia leads to low haemoglobin levels in infected mice, reflecting the anaemia observed in human babesiosis. Similar to humans, B. microti coinfection appears to enhance the severity of Lyme disease-like symptoms in mice. Coinfected mice have lower peak B. microti parasitaemia compared to mice infected with B. microti alone, which may reflect attenuation of babesiosis symptoms reported in some human coinfections. These findings suggest that B. burgdorferi coinfection attenuates parasite growth while B. microti presence exacerbates Lyme disease-like symptoms in mice.


Long-term safety and effectiveness of growth hormone therapy in Korean children with growth disorders: 5-year results of LG Growth Study.
The aim of this registry study was to analyze the long-term safety and effectiveness of recombinant human growth hormone (rhGH) in South Korean pediatric patients (≥2 years of age) with growth hormone deficiency GHD) of idiopathic or organic etiology, idiopathic short stature, Turner syndrome, small for gestational age and chronic renal failure. The study patients were followed-up till two years after the epiphyseal closure, with visits scheduled every six months. The outcome measures included the incidence of adverse events (AEs, in particular, neoplasia, glucose intolerance and hypothyroidism), as well as height standard deviation score (Ht SDS) and annual height velocity. The results of the interim analysis of a 5-year accumulated data for 2,024 patients (7,342 patient-years, PY) are presented. A total of 14 neoplasms were diagnosed (191/100,000 PY); 7 out of 9 malignancies were recurrent craniopharyngioma found in patients with organic GHD. Seven cases of glucose intolerance (95/100,000 PY) and 22 cases of hypothyroidism (300/100,000 PY) were detected; about half of the cases (4 and 10 cases each) were considered to be related with rhGH treatment. Most of the growth-retarded patients showed continuous improvement in Ht SDS, with the most prominent effect observed within a year of treatment initiation. The beneficial effect of rhGH on Ht SDS gain was maintained for 2-4 years. The incidence of AEs of interest in rhGH-treated patients was low, and most of the neoplasms were benign and/or non-related to rhGH. Most patients benefited from the therapy in terms of height increment.


Leukocyte telomere length and age at menopause.
Telomere length is a marker of cellular aging that varies with the individual, is inherited, and is highly correlated across somatic cell types within persons. Interindividual variability of telomere length may partly explain differences in reproductive aging rates. We examined whether leukocyte telomere length was associated with menopausal age. We evaluated the relationship between leukocyte telomere length and age at natural menopause in 486 white women ≥65 years of age. We fit linear regression models adjusted for age, income, education, body mass index, physical activity, smoking, and alcohol intake. We repeated the analysis in women with surgical menopause. We also performed sensitivity analyses excluding women (1) with unilateral oophorectomy, (2) who were nulliparous, or (3) reporting menopausal age <40 years, among other exclusions. For every 1-kb increase in leukocyte telomere length, average age at natural menopause increased by 10.2 months (95% confidence interval = 1.3 to 19.0). There was no association among 179 women reporting surgical menopause. In all but one sensitivity analysis, the association between leukocyte telomere length and age at menopause became stronger. However, when excluding women with menopausal age <40 years, the association decreased to 7.5 months (-0.4 to 15.5). Women with the longest leukocyte telomere length underwent menopause 3 years later than those with the shortest leukocyte telomere length. If an artifact, an association would likely also have been observed in women with surgical menopause. If these results are replicated, leukocyte telomere length may prove to be a useful predictor of age at menopause.


Haplotypes at the Phg-2 Locus Are Determining Pathotype-Specificity of Angular Leaf Spot Resistance in Common Bean.
Angular leaf spot (ALS) is one of the most devastating diseases of common bean (Phaseolus vulgaris L.) and causes serious yield losses worldwide. ALS resistance is reportedly pathotype-specific, but little is known about the efficacy of resistance loci against different pathotypes. Here, we report on ALS resistance evaluations of 316 bean lines under greenhouse and field conditions at multiple sites in Colombia and Uganda. Surprisingly, genome-wide association studies revealed only two of the five previously described resistance loci to be significantly associated with ALS resistance. Phg-2 on chromosome eight was crucial for ALS resistance in all trials, while the resistance locus Phg-4 on chromosome 4 was effective against one particular pathotype. Further dissection of Phg-2 uncovered an unprecedented diversity of functional haplotypes for a resistance locus in common bean. DNA sequence-based clustering identified eleven haplotype groups at Phg-2. One haplotype group conferred broad-spectrum ALS resistance, six showed pathotype-specific effects, and the remaining seven did not exhibit clear resistance patterns. Our research highlights the importance of ALS pathotype-specificity for durable resistance management strategies in common bean. Molecular markers co-segregating with resistance loci and haplotypes will increase breeding efficiency for ALS resistance and allow to react faster to future changes in pathogen pressure and composition.


Gut microbiota dysbiosis and altered tryptophan catabolism contribute to autoimmunity in lupus-susceptible mice.
The autoimmune disease systemic lupus erythematosus (SLE) is characterized by the production of pathogenic autoantibodies. It has been postulated that gut microbial dysbiosis may be one of the mechanisms involved in SLE pathogenesis. Here, we demonstrate that the dysbiotic gut microbiota of triple congenic (TC) lupus-prone mice (B6.Sle1.Sle2.Sle3) stimulated the production of autoantibodies and activated immune cells when transferred into germfree congenic C57BL/6 (B6) mice. Fecal transfer to B6 mice induced autoimmune phenotypes only when the TC donor mice exhibited autoimmunity. Autoimmune pathogenesis was mitigated by horizontal transfer of the gut microbiota between co-housed lupus-prone TC mice and control congenic B6 mice. Metabolomic screening identified an altered distribution of tryptophan metabolites in the feces of TC mice including an increase in kynurenine, which was alleviated after antibiotic treatment. Low dietary tryptophan prevented autoimmune pathology in TC mice, whereas high dietary tryptophan exacerbated disease. Reducing dietary tryptophan altered gut microbial taxa in both lupus-prone TC mice and control B6 mice. Consequently, fecal transfer from TC mice fed a high tryptophan diet, but not a low tryptophan diet, induced autoimmune phenotypes in germfree B6 mice. The interplay of gut microbial dysbiosis, tryptophan metabolism and host genetic susceptibility in lupus-prone mice suggest that aberrant tryptophan metabolism may contribute to autoimmune activation in this disease.


Life-course social influences on tooth loss and oral attitudes among older people: evidence from a prospective cohort study.
This study examined the relationship of trajectories in social condition with oral attitudes and major tooth loss, using the social mobility and accumulation life-course models in a cohort. Whether social-condition inequalities remained stable or changed from 65 yr of age to 70 yr of age was investigated. In 1992, 6,346 inhabitants born in 1942 and residing in two Swedish counties agreed to participate in a prospective survey. Of the participants in 1992, 3,585 (47.6% men) completed questionnaires in 1997, 2002, 2007, and 2012. In line with the social-mobility model, the prevalence of negative oral attitudes and major tooth loss in participants at 65 and 70 yr of age showed a consistent gradient according to social-condition trajectory, whereby it was lowest among those who were persistently high and highest among those who were persistently low, with the upwardly and downwardly mobile categories being intermediate. A linear graded association between the number of periods with disadvantaged social condition and oral health supported the accumulation model. Both the social mobility and accumulation life-course models were supported. Social-condition differentials in negative oral attitudes and tooth loss seem to remain stable or to narrow weakly after the usual age of retirement.


Apolipoprotein E gene and retinal microvascular signs in older people: the Cardiovascular Health Study.
To examine the association between apolipoprotein E (APOE) gene polymorphism and retinal microvascular signs in an older population. Retinal photographs were taken of 2,152 participants (1,831 whites, and 321 African-Americans), aged 69-96 years, who were participating in a population-based study of four United States communities. We used standardized protocols to assess photographs for the presence of retinal microvascular signs (retinopathy, arterio-venous nicking, and focal arteriolar narrowing) and a computer-assisted method to measure retinal vessel diameters. We analyzed DNA extracted from blood samples of participants for common allelic variants of the APOE gene. After adjusting for age, gender, systolic blood pressure, smoking, total serum cholesterol, and other risk factors, we found white participants carrying the epsilon2 and epsilon4 alleles were more likely to have arterio-venous nicking than the epsilon3/epsilon3 homozygotes, with odds ratio (OR) of 1.70 and confidence interval (CI) 95% (1.03-2.83) for the epsilon2 carriers and OR 1.74 (95% CI 1.06-2.84) for the epsilon4 carriers. Among white participants without hypertension, the associations remained significant for the epsilon4 carriers (OR 2.32, 95% CI 1.18-4.57). Whites, normotensive carriers of the epsilon2 allele had significantly narrower retinal arteriolar diameters (adjusted mean arteriolar diameter of 163.5 mum, 95% CI 160.1-167.0, p=0.03) compared to the epsilon3/epsilon3 homozygotes (167.8 mum, 95% CI 166.0-169.6). APOE gene polymorphism was not associated with retinopathy, focal narrowing, or retinal venular diameters in white participants. There were insufficient numbers of African-Americans for separate multivariate analysis. This study provides little evidence that the APOE gene polymorphism plays a significant role in the pathogenesis of retinal microvascular changes in the general population. In the older white population, APOE epsilon2 and epsilon4 allele carriers were more likely to have arterio-venous nicking. Other retinal signs, however, were not related to APOE gene polymorphism.


Environmental dimensionality determines species coexistence.
According to the competitive-exclusion principle, the number n of regulating variables describing a given community dynamics is an upper bound on the number of species (or types or morphs) that can coexist at equilibrium. On occasion, it is possible to reformulate a model with a lower number of regulating variables than appeared in the initial specification. We call the smallest number of such variables the dimension of the environmental feedback, or environmental dimension for short. For studying which species can invade a community, it is enough to know the sign of each species' long-term growth rate, i.e., invasion fitness. Therefore, different indicators of population growth - so-called fitness proxies, such as the basic reproduction number-are sometimes preferred. However, as we show, different fitness proxies may have different dimensions. Fundamental characteristics such as the environmental dimension should not depend on such arbitrary choices. Here, we resolve this difficulty by introducing a refined definition of environmental dimension that focuses on neutral fitness contours. On this basis, we show that this definition of environmental dimension is not only unambiguous, i.e., independent of the choice of fitness proxy, but also constructive, i.e., applicable without needing to assess an infinite number of possible fitness proxies. We then investigate how to determine environmental dimensions by analysing the two components of the environmental feedback: the impact map describing how a community's resident species affect the regulating variables and the sensitivity map describing how population growth depends on the regulating variables. The dimension of the impact map is lower than n when the set of feasible environments is of lower dimension than n, and the dimension of the sensitivity map is lower than n when not all n regulating variables affect the sign of population growth independently. While the minimum of the dimensions of the impact and sensitivity maps provides an upper bound on the environmental dimension, the combined effect of the two maps can result in an even lower environmental dimension, which happens when the sensitivity map is insensitive to some aspects of the impact map's image. To facilitate the applications of the framework introduced here, we illustrate all key concepts with detailed worked examples. In view of these results, we claim that the environmental dimension is the ultimate generalization of the traditional and widely used notions of the "number of regulating variables" or the "number of limiting factors", and is thus the sharpest generally applicable upper bound on the number of species that can robustly coexist in a community.


Peptide-Conjugated PMOs for the Treatment of Myotonic Dystrophy.
Antisense oligonucleotides (ASOs) have shown great therapeutic potential in the treatment of many neuromuscular diseases including myotonic dystrophy 1 (DM1). However, systemically delivered ASOs display poor biodistribution and display limited penetration into skeletal muscle. The conjugation of cell-penetrating peptides (CPPs) to phosphorodiamidate morpholino oligonucleotides (PMOs), a class of ASOs with a modified backbone, can be used to enhance ASO skeletal muscle penetration. Peptide-PMOs (P-PMOs) have been shown to be highly effective in correcting the DM1 skeletal muscle phenotype in both murine and cellular models of DM1 and at a molecular and functional level. Here we describe the synthesis and conjugation of P-PMOs and methods for analyzing their biodistribution and toxicity in the HSA-LR DM1 mouse model and their efficacy both in vitro and in vivo using FISH and RT-PCR splicing analysis.


Harmful algal blooms: A climate change co-stressor in marine and freshwater ecosystems.
Marine and freshwater ecosystems are warming, acidifying, and deoxygenating as a consequence of climate change. In parallel, the impacts of harmful algal blooms (HABs) on these ecosystems are intensifying. Many eutrophic habitats that host recurring HABs already experience thermal extremes, low dissolved oxygen, and low pH, making these locations potential sentinel sites for conditions that will become more common in larger-scale systems as climate change accelerates. While studies of the effects of HABs or individual climate change stressors on aquatic organisms have been relatively common, studies assessing their combined impacts have been rare. Those doing so have reported strong species- and strain-specific interactions between HAB species and climate change co-stressors yielding outcomes for aquatic organisms that could not have been predicted based on investigations of these factors individually. This review provides an ecological and physiological framework for considering HABs as a climate change co-stressor and considers the consequences of their combined occurrence for coastal ecosystems. This review also highlights critical gaps in our understanding of HABs as a climate change co-stressor that must be addressed in order to develop management plans that adequately protect fisheries, aquaculture, aquatic ecosystems, and human health. Ultimately, incorporating HAB species into experiments and monitoring programs where the effects of multiple climate change stressors are considered will provide a more ecologically relevant perspective of the structure and function of marine ecosystems in future, climate-altered systems.


The NIOSH CROPS Demonstration Project: A Study in New York and Virginia with an Emphasis on Youth.
The agriculture, forestry, and fishing (AgFF) industry sector has had the highest rate of work-related deaths over the past several years. Tractors are the single largest source of occupational fatalities in this industry sector, and tractor rollovers are the largest category of tractor fatalities. The risk of death due to tractor rollovers has been found to be reduced considerably when rollover protective structures (ROPS) are used in conjunction with seatbelts. Increasing the number of tractors with ROPS and seatbelts is one way in which tractor rollover fatalities can be mitigated. The NIOSH cost-effective rollover protective structure (CROPS) project was designed as a demonstration project to determine if CROPS, a type of ROPS, could be retrofitted in the field and were acceptable to farmers. To this end, the CROPS project was successful, with 50 tractors being retrofitted with CROPS in the field. All study participants were asked to complete a pretest, test, and posttest regarding the CROPS retrofit demonstration and their knowledge and attitudes toward ROPS. CROPS demonstrators were the participants who retrofitted their tractors with CROPS. Demonstrators were asked to recruit three to five other participants, known as observers, to watch the demonstration. The rationale was to generate interest in ROPS/CROPS among other farmers and community leaders. Overall, 16% of the participants had youth operating tractors on their farms. Participants reported that 44 youth operated tractors on their farms, more than 25% of these young tractor operators were 4 to 10 years old, and half of the youth operating tractors on participant farms were 13 years old or younger. Only one participant group (demonstrators) included individuals who had all of their tractors protected with ROPS/CROPS for young tractor operators (28%), but they accounted for only one farm each among the New York and Virginia demonstrators. The survey question on ROPS importance did not show an overall increase after respondents participated in the CROPS demonstration project. However, one Virginia observer asked to become a Virginia demonstrator and installed a CROPS on his tractor. Additionally, the questions on attitude toward ROPS showed a general increase in mean scores, with the some of the lowest response categories on the follow-up survey not being selected by respondents, indicating a potentially positive impact on participants' safety attitudes toward ROPS. For the safety attitude questions, the majority remained the same. This study indicates there is still much work to be done to reduce the number of deaths in production agriculture due to tractor rollovers. CROPS could be a valuable addition to the efforts of retrofitting ROPS on tractors in order to reduce the number of deaths due to tractor rollovers.


Elastic filaments in skeletal muscle revealed by selective removal of thin filaments with plasma gelsolin.
Muscle needs an elastic framework to maintain its mechanical stability. Removal of thin filaments in rabbit skeletal muscle with plasma gelsolin has revealed the essential features of elastic filaments. The selective removal of thin filaments was confirmed by staining with phalloidin-rhodamine for fluorescence microscopy, examination of arrowhead formation with myosin subfragment 1 by electron microscopy, and analysis by SDS-PAGE. Thin section electron microscopy revealed the elastic fine filaments (approximately 4 nm in diameter) connecting thick filaments and the Z line. After removal of thin filaments, both rigor stiffness and active tension generation were lost, but the resting tension remained. These observations indicate that the thin filament-free fibers maintain a framework composed of the serial connections of thick filaments, the elastic filaments, and the Z line, which gives passive elasticity to the contractile system of skeletal muscle. The resting tension that remained in the thin filament-free fibers was decreased by mild trypsin treatment. The only protein component that was digested in parallel with the decrease in the resting tension and the disappearance of the elastic filaments was alpha-connectin (also called titin 1), which was transformed from the alpha to the beta form (from titin 1 to 2, respectively). Thus, we conclude that the main protein component of the elastic filaments is alpha-connectin (titin 1).


Differential effects of WAY-100135 on the decrease in 5-hydroxytryptamine release induced by buspirone and NAN-190.
1-(2-Methoxyphenyl)-4-[(phthalimido)butyl] piperazine (NAN-190) and 8-[4-[4-(2-pyrimidinyl)-1-piperazinyl]butyl]-8-azaspiro[4.5] decane-7,9-dione (buspirone) are 5-HT1A receptor partial agonists which decrease 5-hydroxytryptamine (5-HT) release in vivo. In order to assess whether these ligands decrease 5-HT release by stimulating 5-HT1A receptors we examined the ability of the selective 5-HT1A receptor antagonist N-tert-butyl 3-4-(2-methoxyphenyl) piperazin-1-yl-2-phenylpropanamide dihydrochloride (WAY-100135) to block their inhibitory effects on 5-HT. NAN-190 (0.1 mg/kg s.c.) and buspirone (1.0 mg/kg s.c.) significantly decreased extracellular levels of 5-HT in hippocampal dialysates. WAY-100135 (10.0 mg/kg s.c.) attenuated the effect of buspirone but had no significant effect on the NAN-190-induced decreased in 5-HT release. These data demonstrate that buspirone is an agonist at the somatodendritic 5-HT1A receptor but that the inhibitory effects of NAN-190 on 5-HT release may be mediated via a mechanism other than, or in addition to, 5-HT1A receptor agonism.


High-energy noncollinear optical parametric-chirped pulse amplification in LBO at 800 nm.
The optical parametric-chirped pulse amplification (OPCPA) based on large-aperture nonlinear optical crystals is promising for implementation of an ultrahigh peak-power laser system of 10 PW and beyond. We demonstrated the highest energy broadband OPCPA at 800 nm, to the best of our knowledge, by using an 80 mm in diameter LiB(3)O(5)(LBO) amplifier, with an output energy of 28.68 J, a bandwidth of 80 nm (FWHM), and conversion efficiency of 25.38%. After compression, a peak power of 0.61 PW with 33.8 fs pulse duration is produced.


The national comparative audit of surgery for nasal polyposis and chronic rhinosinusitis.
This study summarises the results of a National Audit of sino-nasal surgery carried out in England and Wales. It describes patient and operative characteristics as well as patient outcomes up to 36 months after surgery. Prospective cohort study. NHS hospitals in England and Wales. Consecutive patients undergoing surgery for nasal polyposis and/or chronic rhinosinusitis. The total score derived from a 22-item version of the Sino-Nasal Outcome Test (SNOT-22). Lower scores represent better health-related quality of life. A total of 3128 consecutive patients at 87 NHS hospitals were enrolled. There is a large improvement in SNOT-22 scores from the pre-operative period (mean = 42.0) to 3 months after surgery (mean = 25.5). The scores for patients undergoing nasal polypectomy improved from 41.0 before surgery to 23.1 at 3 months after surgery, while the scores for patients undergoing surgery for chronic rhinosinusitis alone improved from 44.2 to 31.2. The SNOT-22 scores reported at 12 and 36 months after surgery were similar to those reported at 3 months. Excessive bleeding occurred in 5% of patients during the operation and in 1% of patients after the operation. Intra-orbital complications were reported in 0.2%. Of those patients undergoing primary surgery for bilateral grade I or II polyposis, 18% had not received a pre-operative course of steroid treatment. At the 36-month follow-up, 11.4% of patients had undergone revision surgery. The audit confirms that sino-nasal surgery is generally safe and effective. There is some evidence that patient selection for surgery could be improved.


Aggression in low functioning children and adolescents with autistic disorder.
Parents, caregivers and mental health professionals have often reported violence and aggression in children or adolescents with autistic disorder. However, most of these observations derived from anecdotal reports, and studies on frequency and characterization of aggression in autism remain limited. Our objective was to better characterize and understand the different types of aggressive behaviors displayed by a large group of individuals with autism in different observational situations. The study was conducted on 74 children and adolescents with autism and 115 typically developing control individuals matched for sex, age and pubertal stage. Other-Injurious Behaviors (OIB) were assessed in three observational situations (parents at home, two caregivers at day-care, a nurse and a child psychiatrist during blood drawing) using validated scales. The frequency of OIB was significantly higher in individuals with autism compared to typically developing control individuals during the blood drawing (23% vs. 0%, P<0 .01). The parents observed significantly less OIB in their children than caregivers (34% vs. 58%, P<0.05). In addition, the most frequent concurrent behaviors occurring just before the appearance of OIB in individuals with autism were anxiety-related behaviors and excitation according to the parental as well as the caregiver observation. The results suggest that in a stressful situation, such as the blood drawing, individuals with autism release their stress through behaviors such as OIB, whereas typically developing individuals regulate and express their stress through cognitive skills such as mental coping strategies, symbolization skills with representation and anticipation of the stressful situation, social interaction and verbal or non-verbal communication. The findings underline also the key role of the environment in assessing OIB and developing therapeutic perspectives, with an individual who modulates his/her behavior according to the environment, and an environment that perceives this behavior and reacts to it with different tolerance thresholds according to the observers.


Differential Effects of DHA- and EPA-Rich Oils on Sleep in Healthy Young Adults: A Randomized Controlled Trial.
Emerging evidence suggests that adequate intake of omega-3 polyunsaturated fatty acids (n-3 PUFAs), which include docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), might be associated with better sleep quality. N-3 PUFAs, which must be acquired from dietary sources, are typically consumed at suboptimal levels in Western diets. Therefore, the current placebo-controlled, double-blind, randomized trial, investigated the effects of an oil rich in either DHA or EPA on sleep quality in healthy adults who habitually consumed low amounts of oily fish. Eighty-four participants aged 25-49 years completed the 26-week intervention trial. Compared to placebo, improvements in actigraphy sleep efficiency (p = 0.030) and latency (p = 0.026) were observed following the DHA-rich oil. However, these participants also reported feeling less energetic compared to the placebo (p = 0.041), and less rested (p = 0.017), and there was a trend towards feeling less ready to perform (p = 0.075) than those given EPA-rich oil. A trend towards improved sleep efficiency was identified in the EPA-rich group compared to placebo (p = 0.087), along with a significant decrease in both total time in bed (p = 0.032) and total sleep time (p = 0.019) compared to the DHA-rich oil. No significant effects of either treatment were identified for urinary excretion of the major melatonin metabolite 6-sulfatoxymelatonin. This study was the first to demonstrate some positive effects of dietary supplementation with n-3 PUFAs in healthy adult normal sleepers, and provides novel evidence showing the differential effects of n-3 PUFA supplements rich in either DHA or EPA. Further investigation into the mechanisms underpinning these observations including the effects of n-3 PUFAs on sleep architecture are required.


How dependencies between successive examples affect on-line learning.
We study the dynamics of on-line learning for a large class of neural networks and learning rules, including backpropagation for multilayer perceptrons. In this paper, we focus on the case where successive examples are dependent, and we analyze how these dependencies affect the learning process. We define the representation error and the prediction error. The representation error measures how well the environment is represented by the network after learning. The prediction error is the average error that a continually learning network makes on the next example. In the neighborhood of a local minimum of the error surface, we calculate these errors. We find that the more predictable the example presentation, the higher the representation error, i.e., the less accurate the asymptotic representation of the whole environment. Furthermore we study the learning process in the presence of a plateau. Plateaus are flat spots on the error surface, which can severely slow down the learning process. In particular, they are notorious in applications with multilayer perceptrons. Our results, which are confirmed by simulations of a multilayer perceptron learning a chaotic time series using backpropagation, explain how dependencies between examples can help the learning process to escape from a plateau.


Functional MRI and Wada determination of language lateralization: a case of crossed dominance.
The Wada test has historically been the conventional procedure for determining language lateralization before neurosurgery. However, functional magnetic resonance imaging (fMRI) offers a less invasive alternative to the Wada procedure. Research indicates that the two techniques used together may provide comparable, and sometimes complementary, information that results in improved prediction of postsurgical language ability. We present a case in which use of fMRI in conjunction with Wada testing provided complementary information about language lateralization before neurosurgical resection of a mesial temporal subependymoma for seizure control in a patient with schizencephaly.


Fruit and vegetable consumption and risk of epithelial ovarian cancer: the European Prospective Investigation into Cancer and Nutrition.
The association between consumption of fruit and vegetables and risk of ovarian cancer is still unclear from a prospective point of view. Female participants (n = 325,640) of the European Prospective Investigation into Cancer and Nutrition study, free of any cancer at baseline, were followed on average for 6.3 years to develop ovarian cancer. During 2,049,346 person-years, 581 verified cases of primary, invasive epithelial ovarian cancer were accrued. Consumption of fruits and vegetables as well as subgroups of vegetables, estimated from validated dietary questionnaires and calibrated thereafter, was related to ovarian cancer incidence in multivariable hazard regression models. Histologic subtype specific analyses were done. Total intake of fruit and vegetables, separately or combined, as well as subgroups of vegetables (fruiting, root, leafy vegetables, cabbages) was unrelated to risk of ovarian cancer. A high intake of garlic/onion vegetables was associated with a borderline significant reduced risk of this cancer. The examination by histologic subtype indicated some differential effects of fruit and vegetable intake on ovarian cancer risk. Overall, a high intake of fruits and vegetables did not seem to protect from ovarian cancer. Garlic/onion vegetables may exert a beneficial effect. The study of the histologic subtype of the tumor warrants further investigation.


Induction of Major Histocompatibility Complex-mismatched Mouse Lung Allograft Acceptance With Combined Donor Bone Marrow: Lung Transplant Using a 12-Hour Nonmyeloablative Conditioning Regimen.
Despite broad and intense conventional immunosuppression, long-term survival after lung transplantation lags behind that for other solid organ transplants, primarily because of allograft rejection. Therefore, new strategies to promote lung allograft acceptance are urgently needed. The purpose of the present study was to induce allograft tolerance with a protocol compatible with deceased donor organ utilization. Using the major histocompatibility complex-mismatched mouse orthotopic lung transplant model, we investigated a conditioning regimen consisting of pretransplant T cell depletion, low-dose total body irradiation and posttransplant (donor) bone marrow, and splenocyte infusion followed by posttransplantation cyclophosphamide. Our results show that C57BL/6 recipients of BALB/c lung allografts undergoing this complete short-duration nonmyeloablative conditioning regimen had durable lung allograft acceptance. Mice that lacked 1 or more components of this regimen exhibited significant graft loss. Mechanistically, animals with lung allograft acceptance had established higher levels of donor chimerism, lymphocyte responses which were attenuated to donor antigens but maintained to third-party antigens, and clonal deletion of donor-reactive host Vβ T cells. Frequencies of Foxp3 T regulatory cells were comparable in both surviving and rejected allografts implying that their perturbation was not a dominant cell-regulatory mechanism. Donor chimerism was indispensable for sustained tolerance, as evidenced by acute rejection of allografts in established chimeric recipients of posttransplantation cyclophosphamide after a chimerism-ablating secondary recipient lymphocyte infusion. Together, these data provide proof-of-concept for establishing lung allograft tolerance with tandem donor bone marrow transplantation using a short-duration nonmyeloablative conditioning regimen and posttransplant cyclophosphamide.


Epigenetic profiles in children with a neural tube defect; a case-control study in two populations.
Folate deficiency is implicated in the causation of neural tube defects (NTDs). The preventive effect of periconceptional folic acid supplement use is partially explained by the treatment of a deranged folate-dependent one carbon metabolism, which provides methyl groups for DNA-methylation as an epigenetic mechanism. Here, we hypothesize that variations in DNA-methylation of genes implicated in the development of NTDs and embryonic growth are part of the underlying mechanism. In 48 children with a neural tube defect and 62 controls from a Dutch case-control study and 34 children with a neural tube defect and 78 controls from a Texan case-control study, we measured the DNA-methylation levels of imprinted candidate genes (IGF2-DMR, H19, KCNQ1OT1) and non-imprinted genes (the LEKR/CCNL gene region associated with birth weight, and MTHFR and VANGL1 associated with NTD). We used the MassARRAY EpiTYPER assay from Sequenom for the assessment of DNA-methylation. Linear mixed model analysis was used to estimate associations between DNA-methylation levels of the genes and a neural tube defect. In the Dutch study group, but not in the Texan study group we found a significant association between the risk of having an NTD and DNA methylation levels of MTHFR (absolute decrease in methylation of -0.33% in cases, P-value = 0.001), and LEKR/CCNL (absolute increase in methylation: 1.36% in cases, P-value = 0.048), and a borderline significant association for VANGL (absolute increase in methylation: 0.17% in cases, P-value = 0.063). Only the association between MTHFR and NTD-risk remained significant after multiple testing correction. The associations in the Dutch study were not replicated in the Texan study. We conclude that the associations between NTDs and the methylation of the MTHFR gene, and maybe VANGL and LEKKR/CNNL, are in line with previous studies showing polymorphisms in the same genes in association with NTDs and embryonic development, respectively.


Monolithic integration of a silicon driver circuit onto a lead lanthanum zirconate titanate substrate for smart spatial light modulator fabrication.
The monolithic integration of N-channel metal-oxide-semiconductor (NMOS) driver circuits in silicon thin films onto a lead lanthanum zirconate titanate (PLZT) substrate is reported. Two integration methods are compared. Both methods result in NMOS transistors that exhibit electrical properties that are close to those of transistors fabricated in bulk silicon. The characteristics of PLZT modulators driven by thin-film transistors are also similar to those of bulk PLZT modulators. These techniques promise new spatial light modulators of high complexity and performance that good-quality silicon and bulk PLZT can offer.


[Pathological hoarding by children and adolescents].
There is a dearth of knowledge and understanding concerning hoarding by children and adolescents. Psychiatrists need to know more about the phenomenon of hoarding since it can be a marker of psychopathology and it sometimes is symptomatic of a psychiatric disorder. To review hoarding from an epidemiological and psychopathological perspective and to discuss it in relation to the developmental aspect of the first object acquisition: the transitional object. We conducted a literature search in PubMed, Medline, PsycINFO and the Cochranedatabase using primarily the search term 'hoarding', but also in combination with the terms: primates, child, adolescent, psych*, klepto*, transitional object, obsessive-compulsive disorder, collecting and attachment. Both animals and humans engage frequently in collecting and hoarding. Up to 60% of normally functioning children and adolescents are involved in collecting. A strong emotional attachment to possessions may be a response to an attachment problem. Hoarding combined with psychopathology is seen in persons of all ages but the prevalence rates for children and adults are unknown. Hoarding is a worrisome type of behaviour which must be regarded as an indication of serious comorbid psychopathology. It can occur either as a symptom of an existing disorder or as a separate disorder. Finally we recommend that hoarding be included in the diagnostic criteria of the dsm and icd.


Prescription medication hoarding and borrowing or sharing behaviours in older residents in the Illawarra, New South Wales, Australia.
To examine prescription medication hoarding and borrowing or sharing (PMHBS) behaviours in older people, particularly which medications are subject to these behaviours and the circumstances that enable these behaviours. A mixed methods triangulation design, using consecutive qualitative (focus groups) and quantitative (survey) methodologies in a convenience sample of people older than 65 years, living independently in the Illawarra region (New South Wales). Focus group participants (n = 28) acknowledged PMHBS behaviours were widespread; however, very few survey respondents (n = 226) admitted to engaging in these behaviours. Main findings in the study were enablers for these behaviours: the prescription medication is considered the same as that prescribed previously; and self-medicating for pain relief. The prevalence of PMHBS behaviours in this study was low, although it was acknowledged such behaviours occurred in the wider community. Sharing strong pain medication and the same prescription medication appeared to be acceptable in this population.


Childhood: a suitable case for treatment?
We examine the contemporary debate on attention deficit hyperactivity disorder, in which concerns about medicalisation and overuse of drug treatments are paramount. We show medicalisation in attention deficit hyperactivity disorder to be a complex issue that requires systematic research to be properly understood. In particular, we suggest that the debate on this disorder might be more productive and less divisive if longitudinal, evidence-based understanding of the harms and benefits of psychiatric diagnosis and misdiagnosis existed, as well as better access to effective, non-drug treatments. If articulation of the values that should guide clinical practice in child psychiatry is encouraged, this might create greater trust and less division.


First Report of Cowpea mild mottle virus in Cowpea and French Bean in Taiwan.
In 2009, more than 50% of vine type French beans were found bearing severe viral symptoms in a vegetable garden in Nantou County, Taiwan. Infected plants were stunted and exhibited pronounced mottling symptoms on their leaves. The symptomatic plants were mechanically inoculated on Chenopodium quinoa and local lesions developed 7 to 10 days after inoculation. The virus source established by back isolation the single lesion from C. quinoa on French beans developed symptoms similar to those found in the field. Host range test showed that this isolate could only infect leguminous plants, including soybean, mung bean, pea, peanut, asparagus bean, cowpea, adzuki bean, and lima bean, but not cucurbitaceous and solanaceous plants. Since only Cucumber mosaic virus (CMV) has been reported in Taiwan to induce similar symptoms in French beans, we tested both the field collected and inoculated French beans by CMV antiserum in ELISA but obtained a negative result. Due to subsequent electron microscopy studies that found potyvirus and carlavirus like particles in the leaf dips of infected French beans, we conducted reverse transcription (RT)-PCR using generic degenerate primers for potyviruses (Hrp5/Pot1 (2) and PotZ/Pot1 (3)) and carlaviruses (Decarla-u2 (5'-TGCACTGARTCMGAYTATGARGCYTT-3' and Decarla-d1 (5'-GCACATRTCRTCVCCDGCAAA-3') previously designed in our lab. No amplification was found from the potyvirus primers, while the carlavirus one gave an expected amplicon of 285 bp, which was found sharing 81% nucleotide sequence identity with the replicase gene of Cowpea mild mottle virus (CpMMV) (GenBank Accession No. FJ560903). A primer pair (CpMMV-CPu: 5'-TTTACTCTTAggTWATggAgTC-3' and CpMMV-CPd: 5'-CCTATTAAAACACACAAHTCAAA-3') was thus designed to amplify the complete coat protein (CP) gene based on the reported CP sequences and obtained an expected 867-bp product from our French bean isolate. This 867-bp sequence (JX020701) was confirmed to have 97.6% amino acid sequence identities with the CP gene of a Puerto Rican CpMMV isolate (GU191840). In a separate survey, another isolate from asparagus bean (CpMMV-V) causing mild mottling symptom was obtained. Analyses of the CP gene of CpMMV-V (JX070669) confirmed that it shared 88.8% and 97.8% of nucleotide and amino acid sequence identities with the French bean isolate, respectively. Different from most carlaviruses with aphid transmissibility, CpMMV has been shown to be transmitted non-persistently by whiteflies (1). Both CpMMV isolates from Taiwan were confirmed to be transmitted by silver leaf whiteflies (Bemicia argentifolii Bellows and Perring). This is the first record of whitefly transmissible legume virus in Taiwan. Since whitefly has been a problem in agriculture worldwide, CpMMV can be a new emerging threat for Taiwan's legume crop production. References: (1) M. Iwaki et al. Plant Dis. 66:365, 1982. (2) S. S. Pappu et al. J. Virol. Methods 41:9, 1993. (3) F. M. Zerbini et al. Phytopathology 85:746.


Intracellular delivery of peptide drugs using viral nanoparticles of bacteriophage P22: covalent loading and cleavable release.
Peptides have great potential in the treatment of various diseases because of their high specificity and safety. However, their application has been limited by the lack of appropriate delivery techniques. In this study, we have demonstrated the intracellular delivery of peptides using viral nanoparticles (VNPs) of bacteriophage P22 through covalent loading of two different peptides with synergistic cytotoxic effects and controllable release triggered by Cathepsin B, a highly over-expressed protease in many tumors. Fluorescence imaging of the intracellular localization of the P22 VNPs and the peptides evidenced that the delivery system could be transported as designed. Cell Counting Kit-8 (CCK-8) assay and flow cytometry analysis revealed the cytotoxic effects of the peptides on MDA-MB-231 tumor cells. These results have confirmed the feasibility of genetically fusing cargo peptides to the capsid protein of P22 and cleavable release thereafter. This study may be instructive for the design of drug delivery systems and multifunctional theranostic devices based on P22 VNPs.


Prevalence and treatment of pain in older adults in nursing homes and other long-term care institutions: a systematic review.
The high prevalence of pain in older adults and its impact in this age group make it a public health issue, yet few studies of pain relief focus on older adults. Residents of long-term care facilities have more cognitive impairment than their community-living counterparts and may have difficulty reporting the presence and severity of pain. This systematic literature review was conducted to determine the prevalence of pain, and the type and effectiveness of interventions that have been used to treat pain in residents of nursing homes. Studies were identified by searching MEDLINE (from January 1966 to May 1997), HEALTH (from January 1975 to May 1997), CINAHL (from January 1982 to April 1997), AGELINE (from January 1978 to April 1997) and the Cochrane Library (1997, issue 1) and by performing a manual search of textbooks and reference lists. Studies of any methodological design were included if they estimated the prevalence of pain in nursing homes or other long-term care institutions or evaluated interventions for the treatment of pain in residents. Of the 14 eligible studies, 12 were noncomparative studies, 1 was a comparison study with nonrandomized contemporaneous controls, and 1 was a randomized controlled trial. Information on several factors was extracted from each study, including study design, number of patients and facilities, main outcomes measured, methods used to identify and detect pain, prevalence and types of pain, and interventions used to treat pain. The strength of the evidence provided by each study was also assessed. In the 6 studies with data from self-reporting or chart reviews, the prevalence of pain ranged from 49% to 83%. In the 5 studies with data on analgesic use only, the prevalence of pain ranged from 27% to 44%. Only 3 studies, with just 30 patients in total, evaluated an intervention for the treatment of pain. Despite the high prevalence of pain in residents of nursing homes, there is a lack of studies evaluating interventions to relieve their pain. The authors make recommendations for future studies in this area.


The correlation between the bone probing, radiographic and histometric measurements of bone level after regenerative surgery.
The most accurate method of assessing bone level is to elevate the flap and measure the bone level directly. However, this method causes discomfort to the patient and can damage the tissues. Therefore, many studies have been conducted to find an alternative method that can be used to assess the bone level clinically with accuracy and reliability. In the present study, we evaluated the clinical reliability and accuracy of bone probing and radiographic measurements, by comparing the bone levels obtained by both of these measurement techniques with the histometrically confirmed bone levels, after four different kinds of regenerative therapy. Twenty-four intrabony defects (4 x 4 mm one-wall intrabony defects) were surgically created bilaterally in the mandibular second and fourth premolars of six beagle dogs. The control group underwent a conventional flap operation. The graft group was treated with calcium phosphate glass only, the guided tissue regeneration group was treated with guided tissue regeneration only, and the graft + guided tissue regeneration group was treated with calcium phosphate glass and guided tissue regeneration. Bone probing and radiographic measurements were performed to assess the bone level 8 weeks after the operation and then the subjects were killed to perform the histometric measurements. The correlation between the bone probing depths and the histometric bone levels, and that between the radiographic bone levels and the histometric bone levels were analyzed by Spearman's rank correlation analysis. The statistical significance with respect to the type of regenerative therapy was analyzed by the Kruskal-Wallis test. The difference between the bone probing depth and the histometric bone level measurements was 0.14, and that between the radiographic bone level and histometric bone level was 0.6. The coefficient of correlation between the bone probing depth and the histometric bone level was 0.90, and that between the radiographic bone level and the histometric bone level was 0.73. The type of regenerative therapy had no significant effect on the difference between the histometric bone level and the other measurements. The results of the present study suggest that the bone probing measurement may be a reliable method for the assessment of the actual bone level following any type of periodontal regenerative therapy.


Effects of Nonpurified and Choline Supplemented or Nonsupplemented Purified Diets on Hepatic Steatosis and Methionine Metabolism in C3H Mice.
Previous studies indicated that nonpurified and purified commercially available control murine diets have different metabolic effects with potential consequences on hepatic methionine metabolism and liver histology. We compared the metabolic and histological effects of commercial nonpurified (13% calories from fat; 57% calories from carbohydrates with 38 grams/kg of sucrose) and purified control diets (12% calories from fat; 69% calories from carbohydrates with ∼500 grams/kg of sucrose) with or without choline supplementation administered to C3H mice with normal lipid and methionine metabolism. Diets were started 2 weeks before mating, continued through pregnancy and lactation, and continued in offspring until 24 weeks of age when we collected plasma and liver tissue to study methionine and lipid metabolism. Compared to mice fed nonpurified diets, the liver/body weight ratio was significantly higher in mice fed either purified diet, which was associated with hepatic steatosis and inflammation. Plasma alanine aminotransferase levels were higher in mice receiving the purified diets. The hepatic S-adenosylmethionine (SAM)/S-adenosylhomocysteine (SAH) ratio was higher in female mice fed purified compared to nonpurified diet (4.6 ± 2 vs. 2.8 ± 1.9; P < 0.05). Choline supplementation was associated with improvement of some parameters of lipid and methionine metabolism in mice fed purified diets. Standard nonpurified and purified diets have significantly different effects on development of steatosis in control mice. These findings can help in development of animal models of fatty liver and in choosing appropriate laboratory control diets for control animals.


Directed evolution of the 3C protease from coxsackievirus using a novel fluorescence-assisted intracellular method.
Proteases are crucial for regulating biological processes in organisms through hydrolysis of peptide bonds. Recombinant proteases have moreover become important tools in biotechnological, and biomedical research and as therapeutics. We have developed a label-free high-throughput method for quantitative assessment of proteolytic activity in Escherichia coli. The screening method is based on co-expression of a protease of interest and a reporter complex. This reporter consists of an aggregation-prone peptide fused to a fluorescent protein via a linker that contains the corresponding substrate sequence. Cleavage of the substrate rescues the fluorescent protein from aggregation, resulting in increased fluorescence that correlates to proteolytic activity, which can be monitored using flow cytometry. In one round of flow-cytometric cell sorting, we isolated an efficiently cleaved tobacco etch virus (TEV) substrate from a 1:100 000 background of non-cleavable sequences, with around 6000-fold enrichment. We then engineered the 3C protease from coxsackievirus B3 (CVB3 3Cpro) towards improved proteolytic activity on the substrate LEVLFQ↓GP. We isolated highly proteolytic active variants from a randomly mutated CVB3 3Cpro library with up to 4-fold increase in activity. The method enables simultaneous measurement of proteolytic activity and protease expression levels and can therefore be applied for protease substrate profiling, as well as directed evolution of proteases.


Failures in communication and information transfer across the surgical care pathway: interview study.
Effective communication is imperative to safe surgical practice. Previous studies have typically focused upon the operating theatre. This study aimed to explore the communication and information transfer failures across the entire surgical care pathway. Using a qualitative approach, semi-structured interviews were conducted with 18 members of the multidisciplinary team (seven surgeons, five anaesthetists and six nurses) in an acute National Health Service trust. Participants' views regarding information transfer and communication failures at each phase of care, their causes, effects and potential interventions were explored. Interviews were recorded, transcribed verbatim, and submitted to emergent theme analysis. Sampling ceased when categorical and theoretical saturation was achieved. Preoperatively, lack of communication between anaesthetists and surgeons was the most common problem (13/18 participants). Incomplete handover from the ward to theatre (12/18) and theatre to recovery (15/18) were other key problems. Work environment, lack of protocols and primitive forms of information transfer were reported as the most common cause of failures. Participants reported that these failures led to increased morbidity and mortality. Healthcare staff were strongly supportive of the view that standardisation and systematisation of communication processes was essential to improve patient safety. This study suggests communication failures occur across the entire continuum of care and the participants opined that it could have a potentially serious impact on patient safety. This data can be used to plan interventions targeted at the entire surgical pathway so as to improve the quality of care at all stages of the patient's journey.


Gout treatment and comorbidities: a retrospective cohort study in a large US managed care population.
Gout prevalence increased in recent years to become one of the most common causes of inflammatory arthritis in most industrialised countries. Comorbidities may affect the disease severity and treatment patterns. We describe the main characteristics of gout patients, gout-related treatment patterns and prevalent comorbidities in a managed care population. From the large US PharMetrics Patient-Centric Database, patients aged 20-89 with at least 2 claims for a diagnosis of gout (ICD9 274.xx) and related prescriptions between January 1, 1996 and December 31, 2008 were included. Gout flares were ascertained during follow-up. Sex-specific multivariable Poisson regression models were used to assess factors associated with number of flares. 177,637 gout patients were included (mean age 55.2 years; men 75.6%). Overall, more than half (58.1%) had any of the considered comorbidities; hypertension (36.1%), dyslipidemia (27.0%) and diabetes (15.1%) being the most common. Nonselective NSAIDs were the most commonly dispensed (in 38.7% of patients). Notably, 39% of patients did not receive any prescription medication for gout. Patients with comorbidities were significantly more likely to receive anti-gout prescriptions. During an acute episode the prescription of NSAIDs and colchicine increased; and 29.9% of patients received allopurinol. The risk of flares was associated with cardiometabolic comorbidities and older age in women (highest at age 60-69), while in men it decreased by age. Women with these conditions were 60% more likely to have flares (incidence rate ratio, IRR 1.60;1.48-1.74), while men were 10% (IRR 1.10; 1.06-1.13) more likely. Comorbidities affected gout treatment patterns and the occurrence and frequency of acute attacks. Cardiometabolic comorbidities, common in this patients' population, were associated with an increased risk of flares.


Association Between Incretin-Based Drugs and the Risk of Acute Pancreatitis.
The association between incretin-based drugs, such as dipeptidyl peptidase 4 (DPP-4) inhibitors and glucagon-like peptide 1 (GLP-1) agonists, and acute pancreatitis is controversial. To determine whether the use of incretin-based drugs, compared with the use of 2 or more other oral antidiabetic drugs, is associated with an increased risk of acute pancreatitis. A large, international, multicenter, population-based cohort study was conducted using combined health records from 7 participating sites in Canada, the United States, and the United Kingdom. An overall cohort of 1 532 513 patients with type 2 diabetes initiating the use of antidiabetic drugs between January 1, 2007, and June 30, 2013, was included, with follow-up until June 30, 2014. Current use of incretin-based drugs compared with current use of at least 2 oral antidiabetic drugs. Nested case-control analyses were conducted including hospitalized patients with acute pancreatitis matched with up to 20 controls on sex, age, cohort entry date, duration of treated diabetes, and follow-up duration. Hazard ratios (HRs) and 95% CIs for hospitalized acute pancreatitis were estimated and compared current use of incretin-based drugs with current use of 2 or more oral antidiabetic drugs. Secondary analyses were performed to assess whether the risk varied by class of drug (DPP-4 inhibitors and GLP-1 agonists) or by duration of use. Site-specific HRs were pooled using random-effects models. Of 1 532 513 patients included in the analysis, 781 567 (51.0%) were male; mean age was 56.6 years. During 3 464 659 person-years of follow-up, 5165 patients were hospitalized for acute pancreatitis (incidence rate, 1.49 per 1000 person-years). Compared with current use of 2 or more oral antidiabetic drugs, current use of incretin-based drugs was not associated with an increased risk of acute pancreatitis (pooled adjusted HR, 1.03; 95% CI, 0.87-1.22). Similarly, the risk did not vary by drug class (DPP-4 inhibitors: pooled adjusted HR, 1.09; 95% CI, 0.86-1.22; GLP-1 agonists: pooled adjusted HR, 1.04; 95% CI, 0.81-1.35) and there was no evidence of a duration-response association. In this large population-based study, use of incretin-based drugs was not associated with an increased risk of acute pancreatitis compared with other oral antidiabetic drugs.


Genome-wide estimation of transcript concentrations from spotted cDNA microarray data.
A method providing absolute transcript concentrations from spotted microarray intensity data is presented. Number of transcripts per microg total RNA, mRNA or per cell, are obtained for each gene, enabling comparisons of transcript levels within and between tissues. The method is based on Bayesian statistical modelling incorporating available information about the experiment from target preparation to image analysis, leading to realistically large confidence intervals for estimated concentrations. The method was validated in experiments using transcripts at known concentrations, showing accuracy and reproducibility of estimated concentrations, which were also in excellent agreement with results from quantitative real-time PCR. We determined the concentration for 10,157 genes in cervix cancers and a pool of cancer cell lines and found values in the range of 10(5)-10(10) transcripts per microg total RNA. The precision of our estimates was sufficiently high to detect significant concentration differences between two tumours and between different genes within the same tumour, comparisons that are not possible with standard intensity ratios. Our method can be used to explore the regulation of pathways and to develop individualized therapies, based on absolute transcript concentrations. It can be applied broadly, facilitating the construction of the transcriptome, continuously updating it by integrating future data.


The size illusion: visual and kinaesthetic information in size perception.
Earlier studies have shown the size of kinaesthetically presented two-dimensional movement patterns to be significantly overestimated. Whether this size overestimation is characteristic of the kinaesthetic system alone has not been established. Two experiments are reported which were designed to investigate size judgment made after kinaesthetic and visual pattern presentation and the effect of environmental cues on the perception of movement patterns. In experiment 1 patterns were presented kinaesthetically (experimenter guided hand movements around the outline of the pattern) or in combination with visual information given by a moving light (pinpoint light attached to the stylus which was moved around the pattern); visual and kinaesthetic cues were either congruent or conflicting with each other; and environmental cues were either present or absent. In experiment 2 static visual display was compared with visually traced pattern presentation, again with or without environmental cues. Overall the results showed that, regardless of experimental manipulation, in all cases where the information was given over time the subject perceived the pattern larger than reality. After static visual display, overestimation of size did not occur.


Prediction of transposable element derived enhancers using chromatin modification profiles.
Experimentally characterized enhancer regions have previously been shown to display specific patterns of enrichment for several different histone modifications. We modelled these enhancer chromatin profiles in the human genome and used them to guide the search for novel enhancers derived from transposable element (TE) sequences. To do this, a computational approach was taken to analyze the genome-wide histone modification landscape characterized by the ENCODE project in two human hematopoietic cell types, GM12878 and K562. We predicted the locations of 2,107 and 1,448 TE-derived enhancers in the GM12878 and K562 cell lines respectively. A vast majority of these putative enhancers are unique to each cell line; only 3.5% of the TE-derived enhancers are shared between the two. We evaluated the functional effect of TE-derived enhancers by associating them with the cell-type specific expression of nearby genes, and found that the number of TE-derived enhancers is strongly positively correlated with the expression of nearby genes in each cell line. Furthermore, genes that are differentially expressed between the two cell lines also possess a divergent number of TE-derived enhancers in their vicinity. As such, genes that are up-regulated in the GM12878 cell line and down-regulated in K562 have significantly more TE-derived enhancers in their vicinity in the GM12878 cell line and vice versa. These data indicate that human TE-derived sequences are likely to be involved in regulating cell-type specific gene expression on a broad scale and suggest that the enhancer activity of TE-derived sequences is mediated by epigenetic regulatory mechanisms.


The Patient Health Questionnaire 12 Somatic Symptom scale as a predictor of symptom severity and consulting behaviour in patients with irritable bowel syndrome and symptomatic diverticular disease.
Anxiety, depression and nongastrointestinal symptoms are often prominent in irritable bowel syndrome (IBS), but their relative value in patient management has not been quantitatively assessed. We modified the Patient Health Questionnaire 15 (PHQ-15) by excluding three gastrointestinal items to create the PHQ-12 Somatic Symptom (PHQ-12 SS) scale. To compare the value of the PHQ-12 SS scale with the Hospital Anxiety and Depression (HAD) scale in predicting symptoms and patient behaviour in IBS and diverticular disease. We compared 151 healthy volunteers (HV), 319 IBS patients and 296 patients with diverticular disease (DD), 113 asymptomatic [ASYMPDD] and 173 symptomatic DD (SYMPDD). Patient Health Questionnaire 12 SS scores for IBS and SYMPDD were significantly higher than HV. Receiver-operator curves showed a PHQ-12 SS >6, gave a sensitivity for IBS of 66.4% with a specificity of 94.7% and a positive likelihood ratio (PLR) = 13.2, significantly better than that associated with an HAD anxiety score >7, PLR = 3.0 and depression score >7 PLR = 6.5. PHQ-12 SS correlated strongly with IBS severity scale and GP visits in both IBS and DD. The PHQ-12 SS scale is a useful clinical tool which correlates with patient behaviour in both IBS and symptomatic DD.


Assessing phagotrophy in the mixotrophic ciliate Paramecium bursaria using GFP-expressing yeast cells.
We used cells of the yeast Saccharomyces cerevisiae expressing green fluorescent protein (GFP) as fluorescently labelled prey to assess the phagocytic activities of the mixotrophic ciliate Paramecium bursaria, which harbours symbiotic Chlorella-like algae. Because of different fluorescence spectra of GFP and algal chlorophyll, ingested GFP-expressing yeast cells can be distinguished from endosymbiotic algal cells and directly counted in individual P. bursaria cells using fluorescence microscopy. By using GFP-expressing yeast cells, we found that P. bursaria altered ingestion activities under different physiological conditions, such as different growth phases or the presence/absence of endosymbionts. Use of GFP-expressing yeast cells allowed us to estimate the digestion rates of live prey of the ciliate. In contrast to the ingestion activities, the digestion rate within food vacuoles was not affected by the presence of endosymbionts, consistent with previous findings that food and perialgal vacuoles are spatially and functionally separated in P. bursaria. Thus, GFP-expressing yeast may provide a valuable tool to assess both ingestion and digestion activities of ciliates that feed on eukaryotic organisms.


Reperfusion therapies and poststroke seizures.
Seizures are not only a frequent complication of stroke but have been associated with an unfavorable functional and vital outcome of patients who have had stroke. Facing a new paradigm of acute standard stroke care, acute symptomatic seizures in this clinical setting deserve to be rethought. Reperfusion therapies, the gold standard treatment for acute ischemic stroke, improve long-term survival and outcome of patients who have had stroke and have been associated both with clinical seizures and the occurrence of epileptiform activity in the electroencephalogram (EEG). This narrative review describes the different physiopathological mechanisms underlying the possible association between reperfusion therapies and seizures, both acute symptomatic seizures and unprovoked seizures, and the current evidence regarding the risk of poststroke seizures in treated patients. It also identifies the gaps in our knowledge to foster future studies in this field. By different mechanisms, reperfusions therapies may have opposing effects on the risk of poststroke seizures. There is a need for a better definition of the specific physiopathology of seizures in clinical practice, as many factors can be recognized. Additionally, most of the current clinical evidence refers to acute symptomatic seizures and not to unprovoked seizures or poststroke epilepsy, and our analysis does not support the existence of a strong association between thrombolysis and poststroke seizures. So far, the impact of reperfusion therapies on the frequency of poststroke seizures is unclear. To study this effect, many clinical challenges must be overcome, including a better and clear operational definition of seizures and stroke characteristics, the standard of stroke and epilepsy care and EEG monitoring, and the degree of reperfusion success. Prospective, high quality, larger, and longer follow-up multicentric studies are urgently needed. Additionally, stroke registries can also prove useful in better elucidate whether there is an association between reperfusion therapies and seizures. This article is part of the Special Issue "Seizures & Stroke".


Linewidth enhancement factor in semiconductor lasers subject to various external optical feedback conditions.
The linewidth enhancement factor α of a semiconductor laser under the influences of optical feedback with different feedback strengths, external cavity lengths, and feedback phases are studied both experimentally and theoretically. The value of α is determined from the minimum of the Hopf bifurcation curve when the laser is subject to both optical feedback and optical injection. In the experiment, a pellicle beamsplitter mounted on a PZT stage placed on a linear translation stage is used as the reflector, where the external cavity length can be adjusted continuously from the long cavity regime to the short cavity regime with phase accuracy. With a moderate feedback strength, α is found to increase as the feedback strength increases. Moreover, while α is insensitive to the feedback phase in the long cavity regime, it can be tuned continuously in the short cavity regime when varying the phase. A normalized variation range of 21.59% is obtained experimentally at an external cavity length of 1.5 cm, which can be further enhanced by shortening the external cavity. To the best of our knowledge, this is the first detailed study of α from the long to the short cavity regime in a semiconductor laser subject to optical feedback. More particularly, the continuous tuning of α under phase variation is demonstrated the first time.


Miklós Bodanszky Award Lecture: Advances in the selective targeting of protein phosphatase-1 and phosphatase-2A with peptides.
Protein phosphatase-1 and phosphatase-2A are two ubiquitously expressed enzymes known to catalyze the majority of dephosphorylation reactions on serine and threonine inside cells. They play roles in most cellular processes and are tightly regulated by regulatory subunits in holoenzymes. Their misregulation and malfunction contribute to disease development and progression, such as in cancer, diabetes, viral infections, and neurological as well as heart diseases. Therefore, targeting these phosphatases for therapeutic use would be highly desirable; however, their complex regulation and high conservation of the active site have been major hurdles for selectively targeting them in the past. In the last decade, new approaches have been developed to overcome these hurdles and have strongly revived the field. I will focus here on peptide-based approaches, which contributed to showing that these phosphatases can be targeted selectively and aided in rethinking the design of selective phosphatase modulators. Finally, I will give a perspective on www.depod.org, the human dephosphorylation database, and how it can aid phosphatase modulator design. © 2017 The Authors. Journal of Peptide Science published by European Peptide Society and John Wiley & Sons Ltd.


Polymorphisms at the innate immune receptor TLR2 are associated with Borrelia infection in a wild rodent population.
The discovery of the key role of Toll-like receptors (TLRs) in initiating innate immune responses and modulating adaptive immunity has revolutionized our understanding of vertebrate defence against pathogens. Yet, despite their central role in pathogen recognition and defence initiation, there is little information on how variation in TLRs influences disease susceptibility in natural populations. Here, we assessed the extent of naturally occurring polymorphisms at TLR2 in wild bank voles (Myodes glareolus) and tested for associations between TLR2 variants and infection with Borrelia afzelii, a common tick-transmitted pathogen in rodents and one of the causative agents of human Lyme disease. Bank voles in our population had 15 different TLR2 haplotypes (10 different haplotypes at the amino acid level), which grouped in three well-separated clusters. In a large-scale capture-mark-recapture study, we show that voles carrying TLR2 haplotypes of one particular cluster (TLR2c2) were almost three times less likely to be Borrelia infected than animals carrying other haplotypes. Moreover, neutrality tests suggested that TLR2 has been under positive selection. This is, to our knowledge, the first demonstration of an association between TLR polymorphism and parasitism in wildlife, and a striking example that genetic variation at innate immune receptors can have a large impact on host resistance.


Disease burden in The Netherlands due to infections with Shiga toxin-producing Escherichia coli O157.
Surveys carried out between 1990 and 2000 indicated that the incidence of STEC O157-associated gastroenteritis in The Netherlands was 1250 cases/year (median), of which 180 visited a general practitioner, 40 are reported and 0.6 are fatal, mainly in the elderly. There are approximately 20 cases of STEC O157-associated haemolytic-uraemic syndrome (HUS) per year, mainly in children. There are 2.5 HUS patients per year who develop end-stage renal disease (ESRD). There are an estimated 2 HUS-related and 0.5 ESRD-related fatalities per year. The mean disease burden associated with STEC O157 in the Dutch population is 116 (90% confidence interval 85-160) Disability Adjusted Life Years (DALYs) per year. Mortality due to HUS (58 DALYs), and ESRD (21 DALYs) and dialysis due to ESRD (21 DALYs) constitute the main determinants of disease burden. Sensitivity analysis indicates that uncertainty associated with model assumptions did not have a major effect on these estimates.


Apparently synonymous substitutions in FGFR2 affect splicing and result in mild Crouzon syndrome.
Mutations of fibroblast growth factor receptor 2 (FGFR2) account for a higher proportion of genetic cases of craniosynostosis than any other gene, and are associated with a wide spectrum of severity of clinical problems. Many of these mutations are highly recurrent and their associated features well documented. Crouzon syndrome is typically caused by heterozygous missense mutations in the third immunoglobulin domain of FGFR2. Here we describe two families, each segregating a different, previously unreported FGFR2 mutation of the same nucleotide, c.1083A>G and c.1083A>T, both of which encode an apparently synonymous change at the Pro361 codon. We provide experimental evidence that these mutations affect normal FGFR2 splicing and document the clinical consequences, which include a mild Crouzon syndrome phenotype and reduced penetrance of craniosynostosis. These observations add to a growing list of FGFR2 mutations that affect splicing and provide important clinical information for genetic counselling of families affected by these specific mutations.


Weekly enzyme replacement therapy may slow decline of renal function in patients with Fabry disease who are on long-term biweekly dosing.
This study was performed to determine whether adult male patients with Fabry disease who demonstrate a continuing decline in renal function despite 2 to 4 yr of conventionally dosed agalsidase alfa therapy (0.2 mg/kg every other week [EOW]) show an improved slope of decline with weekly administration using the same dosage. Eleven (27%) of 41 adult male patients with Fabry disease who participated in long-term agalsidase alfa clinical trials and who had demonstrated a slope of decline in estimated GFR (eGFR) of > or =5 ml/min per 1.73 m(2)/yr while receiving long-term treatment with agalsidase alfa at the currently recommended dosage of 0.2 mg/kg, infused EOW, were enrolled in this open-label, prospective study. Patients were switched from EOW to weekly infusions and followed for an additional 24 mo. Before switching to weekly dosing, eGFR was 53.7 +/- 6.3 ml/min per 1.73 m(2) (mean +/- SEM), and mean rate of change in eGFR was -8.0 +/- 0.8 ml/min per 1.73 m(2)/yr. During the 24-mo follow-up period after switching to weekly dosing, the mean rate of change in eGFR was observed to slow to -3.3 +/- 1.4 ml/min/1.73 m(2)/yr (P = 0.01 versus EOW). After switching to weekly dosing, three patients demonstrated an improvement in eGFR and six patients demonstrated a slowing in the rate of eGFR decline; only two patients failed to improve their eGFR slope. A multiple regression model confirmed that the weekly infusion regimen was the strongest explanatory variable for the change in eGFR (P = 0.0008), with a weaker contribution from the concomitant use of angiotensin converting enzyme inhibitors/angiotensin receptor blockers (P = 0.02). These results suggest that weekly infusions of agalsidase alfa at a dosage of 0.2 mg/kg may be beneficial in the subgroup of patients who have Fabry disease and whose kidney function continues to decline after 2 to 4 yr or more of standard EOW dosing.


The Refractive Status and Vision Profile: evaluation of psychometric properties and comparison of Rasch and summated Likert-scaling.
The psychometric properties of the Refractive Status and Vision Profile (RSVP) questionnaire were evaluated using Rasch analysis. Ninety-one myopic patients from a refractive surgery clinic and general optometric practice completed the RSVP. Rasch analysis of the RSVP ordinal data was performed to examine for unidimensionality and item reduction. The traditional Likert-scoring system was compared with a Rasch-scored RSVP and a reduced item Rasch-scored RSVP. Rasch analysis of the original RSVP showed poor targeting of item difficulty to patient quality of life, items with a ceiling effect and underutilized response categories. Combining the underutilized response scales and removal of redundant and misfitting items improved the internal consistency and targeting of the RSVP, and the reduced 20-item Rasch scored RSVP showed greater relative precision over standard Likert scoring in discriminating between the two subject groups. A Rasch scaled quality of life questionnaire is recommended for use in refractive outcomes research.