Characteristics of the Norwegian Coastal Current during Years with High Recruitment of Norwegian Spring Spawning Herring (Clupea harengus L.).
Norwegian Spring Spawning herring (NSSH) Clupea harengus L. spawn on coastal banks along the west coast of Norway. The larvae are generally transported northward in the Norwegian Coastal Current (NCC) with many individuals utilizing nursery grounds in the Barents Sea. The recruitment to this stock is highly variable with a few years having exceptionally good recruitment. The principal causes of recruitment variability of this herring population have been elusive. Here we undertake an event analysis using data between 1948 and 2010 to gain insight into the physical conditions in the NCC that coincide with years of high recruitment. In contrast to a typical year when northerly upwelling winds are prominent during spring, the years with high recruitment coincide with predominantly southwesterly winds and weak upwelling in spring and summer, which lead to an enhanced northward coastal current during the larval drift period. Also in most peak recruitment years, low-salinity anomalies are observed to propagate northward during the spring and summer. It is suggested that consistent southwesterly (downwelling) winds and propagating low-salinity anomalies, both leading to an enhanced northward transport of larvae, are important factors for elevated recruitment. At the same time, these conditions stabilize the coastal waters, possibly leading to enhanced production and improved feeding potential along the drift route to Barents Sea. Further studies on the drivers of early life history mortality can now be undertaken with a better understanding of the physical conditions that prevail during years when elevated recruitment occurs in this herring stock.


OWL-based nanomasks for preparing graphene ribbons with sub-10 nm gaps.
We report a simple and highly efficient method for creating graphene nanostructures with gaps that can be controlled on the sub-10 nm length scale by utilizing etch masks comprised of electrochemically synthesized multisegmented metal nanowires. This method involves depositing striped nanowires with Au and Ni segments on a graphene-coated substrate, chemically etching the Ni segments, and using a reactive ion etch to remove the graphene not protected by the remaining Au segments. Graphene nanoribbons with gaps as small as 6 nm are fabricated and characterized with atomic force microscopy, scanning electron microscopy, and Raman spectroscopy. The high level of control afforded by electrochemical synthesis of the nanowires allows us to specify the dimensions of the nanoribbon, as well as the number, location, and size of nanogaps within the nanoribbon. In addition, the generality of this technique is demonstrated by creating silicon nanostructures with nanogaps.


Differences in gene expression levels between early and later stages of human lung development are opposite to those between normal lung tissue and non-small lung cell carcinoma.
We, for the first time, directly compared gene expression profiles in human non-small cell lung carcinomas (NSCLCs) and in human fetal lung development. Previously reported correlations of gene expression profiles between lung cancer and lung development, deduced from matching data on mouse development and human cancer, have brought important information, but suffered from different timing of mouse and human gene expression during fetal development and fundamental differences in tumorigenesis in mice and humans. We used the suppression subtractive hybridization technique to subtract cDNAs prepared from human fetal lung samples at weeks 10-12 and 22-24 and obtained a cDNA library enriched in the transcripts more abundant at the later stage. cDNAs sequencing and RT-PCR analysis of RNAs from human fetal and adult lungs revealed 12 differentially transcribed genes: ADH1B, AQP1, FOLR1, SLC34A2, CAV1, INMT, TXNIP, TPM4, ICAM-1, HLA-DRA, EFNA1 and HLA-E. Most of these genes were found up-regulated in mice and rats at later stages than in human lung development. In surgical samples of NSCLC, these genes were down-regulated as compared to surrounding normal tissues and normal lungs, thus demonstrating opposite expression profiles for the genes up-regulated during fetal lung development.


Interlaboratory reliability of Hybrid Capture 2.
We evaluated the interlaboratory reproducibility of the Hybrid Capture 2 (HC2; Digene, Gaithersburg, MD), a test for oncogenic human papillomavirus (HPV) DNA, using data from 4 clinical center (CC) laboratories and the quality control (QC) laboratory participating in the ASCUS (atypical squamous cells of undetermined significance) and LSIL (low-grade squamous intraepithelial lesion) Triage Study (ALTS). Residual liquid cytology specimens were tested routinely throughout the duration of ALTS at CC laboratories, and a stratified (by time in the study) random sample of specimens was retested by the HPV QC laboratory using equivalent protocols. Of the specimens selected (N = 1,175, 5.50% of all specimens obtained), 1,072 (91.23%) had sufficient specimen volume for retesting. The kappa value between all CC laboratories and the HPV QC laboratory was 0.84 (95% confidence interval, 0.78-0.89), with kappa values for individual CCs and the HPV QC laboratory ranging from 0.79 to 0.89. Agreement between test results was lowest among results for women with negative cytologic findings (0.73); among those with equivocal or abnormal cytologic findings, kappa values were 0.80 or more. These data show that HC2 is a reliable test for detecting clinically relevant oncogenic HPV DNA.


Nuclear inclusions of pathogenic ataxin-1 induce oxidative stress and perturb the protein synthesis machinery.
Spinocerebellar ataxia type-1 (SCA1) is caused by an abnormally expanded polyglutamine (polyQ) tract in ataxin-1. These expansions are responsible for protein misfolding and self-assembly into intranuclear inclusion bodies (IIBs) that are somehow linked to neuronal death. However, owing to lack of a suitable cellular model, the downstream consequences of IIB formation are yet to be resolved. Here, we describe a nuclear protein aggregation model of pathogenic human ataxin-1 and characterize IIB effects. Using an inducible Sleeping Beauty transposon system, we overexpressed the ATXN1(Q82) gene in human mesenchymal stem cells that are resistant to the early cytotoxic effects caused by the expression of the mutant protein. We characterized the structure and the protein composition of insoluble polyQ IIBs which gradually occupy the nuclei and are responsible for the generation of reactive oxygen species. In response to their formation, our transcriptome analysis reveals a cerebellum-specific perturbed protein interaction network, primarily affecting protein synthesis. We propose that insoluble polyQ IIBs cause oxidative and nucleolar stress and affect the assembly of the ribosome by capturing or down-regulating essential components. The inducible cell system can be utilized to decipher the cellular consequences of polyQ protein aggregation. Our strategy provides a broadly applicable methodology for studying polyQ diseases.


Patterns of Intrahemispheric EEG Asymmetry in Insomnia Sufferers: An Exploratory Study.
Individuals with insomnia present unique patterns of electroencephalographic (EEG) asymmetry between homologous regions of each brain hemisphere, yet few studies have assessed asymmetry within the same hemisphere. Increase in intrahemispheric asymmetry during rapid eye movement (REM) sleep in good sleepers (GS) and disruption of REM sleep in insomnia sufferers (INS) both point out that this activity may be involved in the pathology of insomnia. The objective of the present exploratory study was to evaluate and quantify patterns of fronto-central, fronto-parietal, fronto-occipital, centro-parietal, centro-occipital and parieto-occipital intrahemispheric asymmetry in GS and INS, and to assess their association with sleep-wake misperception, daytime anxiety and depressive symptoms, as well as insomnia severity. This paper provides secondary analysis of standard EEG recorded in 43 INS and 19 GS for three nights in a sleep laboratory. Asymmetry measures were based on EEG power spectral analysis within 0.3-60 Hz computed between pairs of regions at frontal, central, parietal and occipital derivations. Repeated-measures ANOVAs were performed to assess group differences. Exploratory correlations were then performed on asymmetry and sleep-wake misperception, as well as self-reported daytime anxiety and depressive symptoms, and insomnia severity. INS presented increased delta and theta F3/P3 asymmetry during REM sleep compared with GS, positively associated with depressive and insomnia complaints. INS also exhibited decreased centro-occipital (C3/O1, C4/O2) and parieto-occipital (P3-O1, P4/O2) theta asymmetry during REM. These findings suggest that INS present specific patterns of intrahemispheric asymmetry, partially related to their clinical symptoms. Future studies may investigate the extent to which asymmetry is related to sleep-wake misperception or memory impairments.


Dementia and Alzheimer's Disease among Older Kidney Transplant Recipients.
Older patients with ESRD who receive a kidney transplant (KT) may develop post-KT dementia and Alzheimer's disease (AD) associated with their long-standing kidney disease and/or neurotoxic immunosuppressant agents. To investigate this possibility, we studied 40,918 older (aged ≥55 years) KT recipients (January 1, 1999 to December 31, 2011) linked to Medicare claims through the US Renal Data System. We estimated dementia and AD risk (cumulative incidence) and studied factors associated with these sequelae using competing risks models. We estimated the risk of death-censored graft loss and mortality after developing dementia or the AD subtype of dementia, separately, using adjusted Cox proportional hazards models. Older recipients had a 10-year dementia risk ranging from 5.1% for recipients aged 55-60 years to 17.0% for recipients aged ≥75 years; 10-year AD risk ranged from 1.0% to 6.7%, respectively. The strongest predictors for dementia and AD were older recipient age and pretransplant diabetes. The 10-year graft loss risk was 28.8% for those who did not develop dementia and 43.1% for those who did, and the corresponding mortality risks were 55.7% and 89.9%, respectively. Older recipients with dementia had a 1.52-fold (95% confidence interval, 1.39 to 1.68) increased risk of graft loss and a 2.38-fold (95% confidence interval, 2.26 to 2.49) increased risk of mortality. We observed similar results for AD. We conclude that older KT recipients have a high risk of post-KT dementia and AD, and these sequelae associate with a profound effect on patient and graft survival.


Extracranial venous drainage patterns in patients with multiple sclerosis and healthy controls.
CCSVI hypothesizes an association between impaired extracranial venous drainage and MS. Published sonographic criteria for CCSVI are controversial, and no MR imaging data exist to support the CCSVI hypothesis. Our purpose was to evaluate possible differences in the extracranial venous drainage of MS and healthy controls using both TOF and contrast-enhanced TRICKS MRV. Healthy subjects (n = 20) and patients with MS (n = 19) underwent axial 2D-TOF neck MRV (to assess flattening) and TRICKS MRV (to assess collaterals) at 3T. Two neuroradiologists blinded to cohort status scored IJV flattening and the severity of non-IJV collaterals by using a 4-point qualitative scale (normal = 0, mild = 1, moderate = 2, severe = 3). κ was used to assess reader agreement. Comparisons between groups were performed by using the Wilcoxon rank sum test. The Spearman rank correlation was used to assess the relationship between IJV flattening and collateral scores and, in patients with MS, EDSS scores. The 2 groups were matched for age and sex (MS, 45 ± 8 years, 79% female; healthy controls, 47 ± 10 years, 65% female). Reader agreement for IJV flattening and collateral severity was good (κ = 0.74) and moderate (κ = 0.58), respectively. While IJV flattening was seen in both patients with MS and healthy controls, scores for the patients with MS were significantly higher (P = .002). Despite a trend, there was no significant difference in collateral scores between groups (P = .063). There was a significant positive correlation between flattening and collateral scores (ρ = 0.32, P = .005) and EDSS and flattening scores (ρ = 0.45, P = .004) but not between EDSS and collateral scores (ρ = 0.01, P = .97). These results indicate that patients with MS have greater IJV flattening and a trend toward more non-IJV collaterals than healthy subjects. The role that this finding plays in the pathogenesis or progression of MS, if any, requires further study.


The relationship between age and relaxation of vascular smooth muscle in the rabbit and rat.
Relaxation of rabbit and rat aorta by isoproterenol decreases with increasing age, whereas such responses caused by nitroglycerin or sodium nitrite are not age-dependent. In the present study, we sought to determine whether this relationship also exists in pulmonary arteries and portal veins. As was the case with the aorta, isoproterenol-induced relaxation of pulmonary arteries decreased as the animal aged; relaxation by nitroglycerin was minimally altered. Aging did not influence responses of rabbit and rat portal veins to isoproterenol or nitroglycerin. If the responses of these blood vessels are characteristic of the responses of other vascular smooth muscles to vasodilators, then this study suggests a difference in the manner by which arteries and veins age. We also confirmed that isoproterenol-induced relaxation of rabbit and rat aorta markedly decreases with increasing age and that the responses of rat aorta to nitroglycerin are independent of age. Because of the agonist used to contract the tissues prior to drug-induced relaxation, the results of the first series of experiments with nitroglycerin on rabbit aorta were at variance with our earlier findings. When KCl was used to contract the aortas, the mean effective dose (ED50) obtained for nitroglycerin for tissues from 2-year-old rabbits was 8 to 19 times larger than that obtained from 2-month-old rabbits. This ratio dropped to 4 when the tissues were contracted with histamine. Since KCl and histamine contract rabbit aorta by different mechanisms, this finding suggests that, in addition to a specific loss in beta-receptor activity, increasing age results in an alteration in the contraction-relaxation process of rabbit aortic tissue.


Phage Selection of Bicyclic Peptide Ligands of the Notch1 Receptor.
Precise regulation of the Notch signaling pathway is crucial, as increases or deficiencies in signaling are associated with diseases, including a wide range of cancers. Recent studies have demonstrated that monoclonal antibodies that bind and stabilize the structure of the negative regulatory region (NRR) in the extracellular domain of the Notch receptor can inhibit Notch signaling. In this work, we posed the question whether bicyclic peptides, being around 100-fold smaller than antibodies, can also stabilize the NRR and inhibit Notch signaling. Bicyclic peptides that bind the NRR of human Notch1 were isolated from combinatorial libraries by phage display. Affinity maturation yielded ligands with dissociation constant (Kd ) values as low as 150 nM. The bicyclic peptides increased the melting temperature of the NRR by up to 8 °C, thus substantially stabilizing the protein structure, but they did not inhibit Notch signaling in cellular assays. Although Notch signaling could not be inhibited, this work demonstrates that phage-selected bicyclic peptides can stabilize proteins; this capacity of bicyclic peptides may be exploited for modulating the conformation of other disease targets.


Severe Salt-Losing Syndrome and Hyperkalemia Induced by Adult Nephron-Specific Knockout of the Epithelial Sodium Channel α-Subunit.
Systemic pseudohypoaldosteronism type 1 (PHA-1) is a severe salt-losing syndrome caused by loss-of-function mutations of the amiloride-sensitive epithelial sodium channel (ENaC) and characterized by neonatal life-threatening hypovolemia and hyperkalemia. The very high plasma aldosterone levels detected under hypovolemic or hyperkalemic challenge can lead to increased or decreased sodium reabsorption, respectively, through the Na(+)/Cl(-) cotransporter (NCC). However, the role of ENaC deficiency remains incompletely defined, because constitutive inactivation of individual ENaC subunits is neonatally lethal in mice. We generated adult inducible nephron-specific αENaC-knockout mice (Scnn1a(Pax8/LC1)) that exhibit hyperkalemia and body weight loss when kept on a regular-salt diet, thus mimicking PHA-1. Compared with control mice fed a regular-salt diet, knockout mice fed a regular-salt diet exhibited downregulated expression and phosphorylation of NCC protein, despite high plasma aldosterone levels. In knockout mice fed a high-sodium and reduced-potassium diet (rescue diet), although plasma aldosterone levels remained significantly increased, NCC expression returned to control levels, and body weight, plasma and urinary electrolyte concentrations, and excretion normalized. Finally, shift to a regular diet after the rescue diet reinstated the symptoms of severe PHA-1 syndrome and significantly reduced NCC phosphorylation. In conclusion, lack of ENaC-mediated sodium transport along the nephron cannot be compensated for by other sodium channels and/or transporters, only by a high-sodium and reduced-potassium diet. We further conclude that hyperkalemia becomes the determining factor in regulating NCC activity, regardless of sodium loss, in the ENaC-mediated salt-losing PHA-1 phenotype.


Regulation of cell wall synthesis and growth.
The replication and growth of Mycobacterium tuberculosis are fundamentally linked to the synthesis and extension of its complex cell wall. Incorporation of new wall material must be tightly regulated so that its deposition does not compromise the extant structure. M. tuberculosis also produces an impressive array of complex bioactive lipids that are intimately involved in pathogenesis and protective immunity. The profiles of these lipids are regulated appropriately to allow the bacterium to respond to the prevailing conditions it faces in vivo. A number of regulatory strategies employed by M. tuberculosis to control cell wall biosynthesis and cell division have now been elucidated. The review highlights the role of alternative sigma factors with extracytoplasmic function in the activation of genes for biosynthesis of complex lipids involved in pathogenicity. Rel(Mtb) and CRP(Mt) play roles in cell wall responses to general nutrient deprivation by synthesis and sensing of starvation second messengers, respectively. Recently, the importance of protein phosphorylation networks in cell wall biosynthesis has attracted considerable interest. A plethora of two-component and eukaryotic-like serine/threonine protein kinases systems have been discovered and several are implicated in cell-division, morphogenesis and regulation of the profile of complex bioactive lipids elaborated by the pathogen.


Solid-Phase Extraction (SPE): Principles and Applications in Food Samples.
Solid-Phase Extraction (SPE) is a sample preparation method that is practised on numerous application fields due to its many advantages compared to other traditional methods. SPE was invented as an alternative to liquid/liquid extraction and eliminated multiple disadvantages, such as usage of large amount of solvent, extended operation time/procedure steps, potential sources of error, and high cost. Moreover, SPE can be plied to the samples combined with other analytical methods and sample preparation techniques optionally. SPE technique is a useful tool for many purposes through its versatility. Isolation, concentration, purification and clean-up are the main approaches in the practices of this method. Food structures represent a complicated matrix and can be formed into different physical stages, such as solid, viscous or liquid. Therefore, sample preparation step particularly has an important role for the determination of specific compounds in foods. SPE offers many opportunities not only for analysis of a large diversity of food samples but also for optimization and advances. This review aims to provide a comprehensive overview on basic principles of SPE and its applications for many analytes in food matrix.


Endovascular treatment of cerebral aneurysms: An in vitro study with detachable platinum coils and tricellulose acetate polymer.
The purpose of our experimental study was to determine the effectiveness of filling the cavity of in vitro aneurysms with detachable platinum coils and the combination of detachable platinum coils and liquid embolic agent. Silicone aneurysm models were connected to a circulatory system to simulate arterial flow. A microcatheter was used to introduce detachable coils into the aneurysm cavities. First, platinum coils were introduced until the point of minimal dense packing, indicated by aneurysmal circulatory exclusion. Packing was continued up to maximal dense packing, indicated by protrusion of the coil into the parent artery. Volumetric ratios (coil volume-aneurysm volume) were calculated for minimal and maximal dense packing. Then, after purposeful undercoiling of aneurysm models, a micropump system was used to fill the aneurysm by stepwise injection of tricellulose acetate polymer through the coil mesh until angiographic aneurysm exclusion was completed. The volumetric ratios of maximal packing with coils and tricellulose acetate polymer in relation to the aneurysm volume were calculated. Maximal dense packing ratios with coils (mean, 32.5%; standard deviation [SD], 3%) were slightly higher than those with the minimal dense packing (mean, 28. 2%; SD, 3%) but were always less than 37%. The ratios of packing with the combined use of coils and tricellulose acetate polymer were greater than 100% (mean, 124.4%; SD, 15%). Knowledge of the volumetric ratio of maximal dense packing was useful for effective filling with coils and tricellulose acetate polymer. The combined use of coils and liquid polymer appeared more effective than the use of coils alone for the complete occlusion of the aneurysm lumen.


Use of chimeric muscarinic receptors to investigate epitopes involved in allosteric interactions.
All five (m1-m5) muscarinic receptors are sensitive to allosteric regulation, but gallamine is considerably more potent in slowing the dissociation of N-[3H]methylscopolamine (NMS) from the m2 subtype than from the m3 or m5 subtypes. To study the structural basis for the preference of gallamine for the m2 subtype, we evaluated [3H]NMS-gallamine interactions with chimeric receptors in which segments of the m5 receptor were systematically replaced with the corresponding m2 sequence. Substitutions that included the sixth transmembrane domain and third extracellular loop resulted in marked increases in the potency of gallamine, but substitutions that did not include these regions were without effect. A similar substitution was investigated using m2/m3 chimeric receptors, in which a segment extending from the middle of the sixth transmembrane domain to the carboxyl terminus was exchanged. As with the m2/m5 constructs, substitution of the m2 carboxyl-terminal segment into the m3 subtype significantly increased the potency of gallamine. Furthermore, the converse substitution reduced the potency of gallamine dramatically, to approximately that seen for the m3 subtype itself. It appears that this portion of the receptor is a critical determinant for the binding of gallamine and/or the allosteric interactions between gallamine and [3H]NMS.


Polymorphism in the third intron of the interferonγ gene is associated with susceptibility to multiple sclerosis.
The present study aims to examine the relationship between polymorphisms in the third intron of the IFN-γ gene and their influence on susceptibility to multiple sclerosis. A population-based case-control study was used for this purpose. Multiple sclerosis patients and healthy controls were interviewed. Genetic polymorphisms of IFN-γ intron III at the +2118 A/G and +3586 G/ACT sites were detected using polymerase chain reaction-restriction fragment length polymorphism. Genotypes and allele frequencies of IFN-γ intron III at the +2118 position were significantly different between multiple sclerosis patients and controls (P ≥ 0.05). However, no difference in allele frequencies was observed at the +3586 position between the two groups (P ≤ 0.05). Thus, polymorphisms at the +2118 A/G site in the IFN-γ intron III gene may be associated with susceptibility to multiple sclerosis.


Theoretical and computational strategies for rational molecularly imprinted polymer design.
The further evolution of molecularly imprinted polymer science and technology necessitates the development of robust predictive tools capable of handling the complexity of molecular imprinting systems. A combination of the rapid growth in computer power over the past decade and significant software developments have opened new possibilities for simulating aspects of the complex molecular imprinting process. We present here a survey of the current status of the use of in silico-based approaches to aspects of molecular imprinting. Finally, we highlight areas where ongoing and future efforts should yield information critical to our understanding of the underlying mechanisms sufficient to permit the rational design of molecularly imprinted polymers.


Gestational Trophoblastic Disease at Sultan Qaboos University Hospital: Prevalence, Risk Factors, Histological Features, Sonographic Findings, and Outcomes.
We sought to assess the prevalence of gestational trophoblastic diseases (GTD) among pregnant women at Sultan Qaboos University Hospital (SQUH) and compare our results with the international studies. We also sought to determine the risk factors, histological features, sonographic findings, and outcomes in women with GTD. We conducted a retrospective cohort study of all women diagnosed with GTD and followed at SQUH between November 2007 and October 2015. We collected data on maternal demographics, risk factors, sonographic features, histological diagnosis, follow-up period, and chemotherapy treatment from the hospital information system. Sixty-four women with GTD were included in the study with a mean age of 31.0±7.5 years, mean gravidity 4.0, and parity 2.0. The prevalence of GTD was 0.3% (one in 386 births), and the most common risk factors were increased maternal age and multiparity. A partial hydatidiform mole was diagnosed in 54.7%, complete hydatidiform mole in 43.8%, and invasive mole in 1.6% of women. Eleven percent of women required chemotherapy. Typical ultrasound features for partial molar pregnancy were present in 54.7% of our sample, while snowstorm appearance was seen in 89.3% of those with complete mole. Negative beta-human chorionic gonadotropin was achieved 70 days after diagnosis in 41 women. The awareness of the risks and complications of GTD among physicians with close follow-up is paramount. There is a need to establish a national registry of GTD cases in Oman.


A longitudinal study using latent curve models of groups with mild cognitive impairment and Alzheimer's disease.
This study explores how mild cognitive impairment (MCI) and Alzheimer's disease (AD) develop over time. NEW METHOD: this study involves a new application of latent curve models (LCM) to examine the development trajectory of a healthy, MCI, and AD groups on a series of clinical and neural measures. Multiple-group latent curve models were used to compare the parameters of the trajectories across groups. LCM results showed that a linear functional form of growth was adequate for all the clinical and neural measures. Positive and significant differences in initial levels were seen across groups on all of the clinical and neural measures. In all groups, the following measures increased slightly, or considerably, over time: Clinical Dementia Rating, Alzheimer's disease Cognitive Assessment, and Montreal Assessment Test for Dementia. In contrast, a slight or a greatly decreasing trajectory was observed on the following measures: Fluorodeoxyglucose, Mini-Mental State Exam, Rey Auditory Verbal Learning Test as well as Hippocampus, Fusiform and Entorhinal Cortex volume measures. However, a constant mean trajectory was seen on Cognition Self Report Memory and languages scores. COMPARISION WITH EXISTING METHODS: there are no prior studies that applied LCM on large AD datasets. cognitive decline occurs in the cognitively normal (CN), MCI, and AD groups but at different rates. Further, some important cognitive, neural, and clinical variables that (a) best differentiate between CN, MCI, and AD as well as (b) differentially change over time in MCI and AD, which may explain disease progression.


Glyphosate-based herbicides are toxic and endocrine disruptors in human cell lines.
Glyphosate-based herbicides are the most widely used across the world; they are commercialized in different formulations. Their residues are frequent pollutants in the environment. In addition, these herbicides are spread on most eaten transgenic plants, modified to tolerate high levels of these compounds in their cells. Up to 400 ppm of their residues are accepted in some feed. We exposed human liver HepG2 cells, a well-known model to study xenobiotic toxicity, to four different formulations and to glyphosate, which is usually tested alone in chronic in vivo regulatory studies. We measured cytotoxicity with three assays (Alamar Blue, MTT, ToxiLight), plus genotoxicity (comet assay), anti-estrogenic (on ERalpha, ERbeta) and anti-androgenic effects (on AR) using gene reporter tests. We also checked androgen to estrogen conversion by aromatase activity and mRNA. All parameters were disrupted at sub-agricultural doses with all formulations within 24h. These effects were more dependent on the formulation than on the glyphosate concentration. First, we observed a human cell endocrine disruption from 0.5 ppm on the androgen receptor in MDA-MB453-kb2 cells for the most active formulation (R400), then from 2 ppm the transcriptional activities on both estrogen receptors were also inhibited on HepG2. Aromatase transcription and activity were disrupted from 10 ppm. Cytotoxic effects started at 10 ppm with Alamar Blue assay (the most sensitive), and DNA damages at 5 ppm. A real cell impact of glyphosate-based herbicides residues in food, feed or in the environment has thus to be considered, and their classifications as carcinogens/mutagens/reprotoxics is discussed.


Correlates of obesity in young black and white women: the CARDIA Study.
Although differences in obesity between Blacks and Whites are well documented in adult women, less information is available on potential correlates of these differences, especially in young adults. The association between behavioral and demographic factors and body size was assessed in 2801 Black and White women aged 18 to 30 years. Black women had significantly higher age-adjusted mean body mass index and subscapular skinfold thickness than did White women. Obesity had different associations with age and education across racial groups. A positive relationship between age and obesity was seen in Black women but not in White women, whereas a negative association between education and body size was noted only in White women. Potential contributing factors to the increased prevalence of obesity in Black women include a more sedentary lifestyle, higher energy intake, earlier menarche, and earlier age at first childbirth. The difference in obesity across race could not be explained completely by these factors, since within virtually all strata, Black women had higher body mass indexes. Further investigation is needed to develop interventional strategies to prevent or reduce excess levels of obesity in Black women.


Carers' experiences of assistive technology use in dementia care: a cross sectional survey.
Assistive Technology (AT) supports persons with dementia and their carers (family, friends and neighbours), yet little is known about experiences and the impact of AT on carers. We report on an exploratory survey that examined the types, uses, costs and impact of AT on carers as well as their quality of life. A cross-sectional survey using the Carers Assistive Technology Experience Questionnaire collected data from carers in the UK, who used at least one AT in the previous year and provided more than 10 h of care for a person with dementia, living at home. Carers completed the questionnaire online or on paper and information on AT, socio-demographic details, and Short-Form Health Survey (SF-12) data were collected. Descriptive and inferential statistics were used to report results and draw conclusions. Data from 201 carers was analysed. Smartphones and tablet computers were the most frequently used AT. AT were used predominantly for safety, communication, and reminders. Carers usually make decisions on buying and continued use of AT. Multiple AT devices were used in the care of persons with dementia and number of AT used was associated with perceived satisfaction. Satisfaction with AT was not related to age, living arrangements and relationship of carers. From the SF-12, Mean Physical Component Score was 49.19 (95%CI- 47.75 to 50.63) and Mental Component Score was 45.37 (95%CI- 43.93 to 46.80). Women, carers in the 46-65 age group and carers who were not extremely satisfied with AT had lower MCS scores. Carers who lived with the person with dementia and older carers had lower PCS scores. Carers report that AT has a beneficial impact. Carers use multiple ATs, perceive AT to be satisfactory and recommend AT use to others. To support carers, we recommend establishment of centrally funded information sources and a loan store for AT. Further research on incremental addition of AT and changes to formal/paid care because of using AT should be undertaken. Practitioners, academics, manufactures and policy makers should consider the experiences of carers in research, development and use of AT to facilitate improved community living of people with dementia.


Normal tubular regeneration and differentiation of the post-ischemic kidney in mice lacking vimentin.
Proliferation and dedifferentiation of tubular cells are the hallmark of early regeneration after renal ischemic injury. Vimentin, a class III intermediate filament expressed only in mesenchymal cells of mature mammals, was shown to be transiently expressed in post-ischemic renal tubular epithelial cells. Vimentin re-expression was interpreted as a marker of cellular dedifferentiation, but its role in tubular regeneration after renal ischemia has also been hypothesized. This role was evaluated in mice bearing a null mutation of the vimentin gene. Expression of vimentin, proliferating cell nuclear antigen (a marker of cellular proliferation), and villin (a marker of differentiated brush-border membranes) was studied in wild-type (Vim+/+), heterozygous (Vim+/-), and homozygous (Vim-/-) mice subjected to transient ischemia of the left kidney. As expected, vimentin was detected by immunohistochemistry at the basal pole of proximal tubular cells from post-ischemic kidney in Vim+/+ and Vim+/- mice from day 2 to day 28. The expression of the reporter gene beta-galactosidase in Vim+/- and Vim-/- mice confirmed the tubular origin of vimentin. No compensatory expression of keratin could be demonstrated in Vim-/- mice. The intensity of proliferating cell nuclear antigen labeling and the pattern of villin expression were comparable in Vim-/-, Vim+/- and Vim+/+ mice at any time of the study. After 60 days, the structure of post-ischemic kidneys in Vim-/- mice was indistinguishable from that of normal non-operated kidneys in Vim+/+ mice. In conclusion, 1) the pattern of post-ischemic proximal tubular cell proliferation, differentiation, and tubular organization was not impaired in mice lacking vimentin and 2) these results suggest that the transient tubular expression of vimentin is not instrumental in tubular regeneration after renal ischemic injury.


Definitions, measurement and indicator selection for quality of care in abortion.
Little consensus exists about how to measure quality of care in abortion. Our purpose is to (a) provide common language for healthcare quality definitions, frameworks and measurement; (b) synthesize literature about quality measurement in abortion; and (c) present criteria for quality metric development. Quality includes effectiveness, patient centeredness, timeliness, efficiency and equity of care. Information about structure, process and outcomes of care is used to measure quality. We do not have good evidence about expected population-level health and behavioral outcomes associated with improving abortion service quality. Abortion patients overwhelmingly report high satisfaction with services, but it is not clear if their satisfaction indicates high-quality care. Guidance exists for quality metric selection; measures must focus on priority topics, be scientifically sound and be feasible. Technical quality standards and clinical guidelines exist, but we lack a standard set of quality metrics. Partners in the Abortion Service Quality Initiative (https://asq-initiative.org/) are collaborating to develop the first-ever global standard for measuring abortion service quality in low- and middle-income countries, both in and out of health care facilities. Standardized and validated quality metrics would move our field forward and contribute to quality improvement activities and, ultimately, to improved health outcomes for women and families. IMPLICATIONS: We define quality of health care, synthesize the evidence about quality of care in abortion and advocate for standardized and validated quality metrics to improve health outcomes for women.


Thermal acclimation has limited effect on the thermal tolerances of summer-collected Arctic and sub-Arctic wolf spiders.
High-latitude ectotherms contend with large daily and seasonal temperature variation. Summer-collected wolf spiders (Araneae; Lycosidae) from sub-Arctic and Arctic habitats have been previously documented as having low temperature tolerance insufficient for surviving year-round in their habitat. We tested two competing hypotheses: that they would have broad thermal breadth, or that they would use plasticity to extend the range of their thermal performance. We collected Pardosa moesta and P. lapponica from the Yukon Territory, Canada, P. furcifera, P. groenlandica, and P. hyperborea from southern Greenland, and P. hyperborea from sub-Arctic Norway, and acclimated them to warm (12 or 20 °C) or cool (4 °C) conditions under constant light for one week. We measured critical thermal minimum (CTmin) or supercooling point (SCP) as a measure of lower thermal limit, and critical thermal maximum (CTmax) as a measure of upper thermal limit. We found relatively little impact of acclimation on thermal limits, and some counterintuitive responses; for example, warm acclimation decreased the SCP and/or cool acclimation increased the CTmax in several cases. Together, this meant that acclimation did not appear to modify the thermal breadth, which supports our first hypothesis, but allows us to reject the hypothesis that spiders use plasticity to fine-tune their thermal physiology, at least in the summer. We note that we still cannot explain how these spiders withstand the very cold winters, and speculate that there are acclimatisation cues or processes that we were unable to capture in our study.


Catechol-based layer-by-layer assembly of composite coatings: a versatile platform to hierarchical nano-materials.
Inspired by the marine mussel's ability to adhere to surfaces underwater, an aqueous catechol-based dip coating platform was developed. Using a catechol-functionalized polyacrylamide binder in combination with inorganic nanoparticles enables the facile fabrication of robust composite coatings via a layer-by-layer process. This modular assembly of well-defined building blocks provides a versatile alternative to electrostatic driven approaches with layer thickness and refractive indices being readily tunable. The platform nature of this approach enables the fabrication of hierarchically ordered nano-materials such as Bragg stacks.


Transforming Obesity Prevention for CHILDren (TOPCHILD) Collaboration: protocol for a systematic review with individual participant data meta-analysis of behavioural interventions for the prevention of early childhood obesity.
Behavioural interventions in early life appear to show some effect in reducing childhood overweight and obesity. However, uncertainty remains regarding their overall effectiveness, and whether effectiveness differs among key subgroups. These evidence gaps have prompted an increase in very early childhood obesity prevention trials worldwide. Combining the individual participant data (IPD) from these trials will enhance statistical power to determine overall effectiveness and enable examination of individual and trial-level subgroups. We present a protocol for a systematic review with IPD meta-analysis to evaluate the effectiveness of obesity prevention interventions commencing antenatally or in the first year after birth, and to explore whether there are differential effects among key subgroups. Systematic searches of Medline, Embase, Cochrane Central Register of Controlled Trials, Cumulative Index to Nursing and Allied Health Literature (CINAHL), PsycInfo and trial registries for all ongoing and completed randomised controlled trials evaluating behavioural interventions for the prevention of early childhood obesity have been completed up to March 2021 and will be updated annually to include additional trials. Eligible trialists will be asked to share their IPD; if unavailable, aggregate data will be used where possible. An IPD meta-analysis and a nested prospective meta-analysis will be performed using methodologies recommended by the Cochrane Collaboration. The primary outcome will be body mass index z-score at age 24±6 months using WHO Growth Standards, and effect differences will be explored among prespecified individual and trial-level subgroups. Secondary outcomes include other child weight-related measures, infant feeding, dietary intake, physical activity, sedentary behaviours, sleep, parenting measures and adverse events. Approved by The University of Sydney Human Research Ethics Committee (2020/273) and Flinders University Social and Behavioural Research Ethics Committee (HREC CIA2133-1). Results will be relevant to clinicians, child health services, researchers, policy-makers and families, and will be disseminated via publications, presentations and media releases. CRD42020177408.


Postoperative Neck Ultrasonography Surveillance After Thyroidectomy in Patients With Medullary Thyroid Carcinoma: A Multicenter Study.
For detecting tumor recurrence of medullary thyroid carcinoma (MTC) in the neck, an appropriate frequency and interval of postoperative ultrasonography (US) surveillance remains unclear. This study aimed to assess an appropriate interval and frequency of postoperative neck US surveillance for detecting tumor recurrence in patients who had undergone thyroid surgery due to MTC. A total of 86 patients who had undergone thyroid surgery for the treatment of MTC and had at least one postoperative US follow-up examination at any of nine affiliated hospitals were included. Postoperative follow-up US, clinical, and histopathological results of patients were reviewed. The tumor recurrence/persistence rate of MTC was investigated, and the interval and session number of postoperative follow-up US and clinicopathologic factors were compared between tumor recurrence/persistence and non-recurrence groups. Of the 86 patients, 22 (25.6%) showed tumor recurrence/persistence. Of the 22 patients with tumor recurrence/persistence, 11 (50%) showed structural recurrence/persistence in the neck on follow-up US. In these 11 patients, the mean interval and session number of postoperative follow-up US between initial surgery and the first US detection of recurrence/persistence was 41.3 ± 39.3 months (range, 6-128 months) and 2.6 ± 2.3 (range, 1-8), respectively. On follow-up US, 6 (54.5%, 6/11) were diagnosed with tumor recurrence/persistence within 3 years of the initial surgery. Tumor recurrence/persistence was significantly correlated with TNM stage (p < 0.001) and multiplicity/bilaterality (p = 0.013). For detecting MTC recurrence/persistence, postoperative US surveillance at 1-year intervals may be sufficient within the first 3 years after thyroid surgery, but depending on the presence of relevant risk factors, annual or biannual US surveillance may be recommendable for 4-10 years after thyroid surgery.


GronOR: Massively parallel and GPU-accelerated non-orthogonal configuration interaction for large molecular systems.
GronOR is a program package for non-orthogonal configuration interaction calculations for an electronic wave function built in terms of anti-symmetrized products of multi-configuration molecular fragment wave functions. The two-electron integrals that have to be processed may be expressed in terms of atomic orbitals or in terms of an orbital basis determined from the molecular orbitals of the fragments. The code has been specifically designed for execution on distributed memory massively parallel and Graphics Processing Unit (GPU)-accelerated computer architectures, using an MPI+OpenACC/OpenMP programming approach. The task-based execution model used in the implementation allows for linear scaling with the number of nodes on the largest pre-exascale architectures available, provides hardware fault resiliency, and enables effective execution on systems with distinct central processing unit-only and GPU-accelerated partitions. The code interfaces with existing multi-configuration electronic structure codes that provide optimized molecular fragment orbitals, configuration interaction coefficients, and the required integrals. Algorithm and implementation details, parallel and accelerated performance benchmarks, and an analysis of the sensitivity of the accuracy of results and computational performance to thresholds used in the calculations are presented.


Lamellipodin tunes cell migration by stabilizing protrusions and promoting adhesion formation.
Efficient migration on adhesive surfaces involves the protrusion of lamellipodial actin networks and their subsequent stabilization by nascent adhesions. The actin-binding protein lamellipodin (Lpd) is thought to play a critical role in lamellipodium protrusion, by delivering Ena/VASP proteins onto the growing plus ends of actin filaments and by interacting with the WAVE regulatory complex, an activator of the Arp2/3 complex, at the leading edge. Using B16-F1 melanoma cell lines, we demonstrate that genetic ablation of Lpd compromises protrusion efficiency and coincident cell migration without altering essential parameters of lamellipodia, including their maximal rate of forward advancement and actin polymerization. We also confirmed lamellipodia and migration phenotypes with CRISPR/Cas9-mediated Lpd knockout Rat2 fibroblasts, excluding cell type-specific effects. Moreover, computer-aided analysis of cell-edge morphodynamics on B16-F1 cell lamellipodia revealed that loss of Lpd correlates with reduced temporal protrusion maintenance as a prerequisite of nascent adhesion formation. We conclude that Lpd optimizes protrusion and nascent adhesion formation by counteracting frequent, chaotic retraction and membrane ruffling.This article has an associated First Person interview with the first author of the paper.


Karnal Bunt: A Re-Emerging Old Foe of Wheat.
Wheat (Triticum aestivum L.) crop health assumes unprecedented significance in being the second most important staple crop of the world. It is host to an array of fungal pathogens attacking the plant at different developmental stages and accrues various degrees of yield losses owing to these. Tilletia indica that causes Karnal bunt (KB) disease in wheat is one such fungal pathogen of high quarantine importance restricting the free global trade of wheat besides the loss of grain yield as well as quality. With global climate change, the disease appears to be shifting from its traditional areas of occurrence with reports of increased vulnerabilities of new areas across the continents. This KB vulnerability of new geographies is of serious concern because once established, the disease is extremely difficult to eradicate and no known instance of its complete eradication using any management strategy has been reported yet. The host resistance to KB is the most successful as well as preferred strategy for its mitigation and control. However, breeding of KB resistant wheat cultivars has proven to be not so easy, and the low success rate owes to the scarcity of resistance sources, extremely laborious and regulated field screening protocols delaying identification/validation of putative resistance sources, and complex quantitative nature of resistance with multiple genes conferring only partial resistance. Moreover, given a lack of comprehensive understanding of the KB disease epidemiology, host-pathogen interaction, and pathogen evolution. Here, in this review, we attempt to summarize the progress made and efforts underway toward a holistic understanding of the disease itself with a specific focus on the host-pathogen interaction between T. indica and wheat as key elements in the development of resistant germplasm. In this context, we emphasize the tools and techniques being utilized in development of KB resistant germplasm by illuminating upon the genetics concerning the host responses to the KB pathogen including a future course. As such, this article could act as a one stop information primer on this economically important and re-emerging old foe threatening to cause devastating impacts on food security and well-being of communities that rely on wheat.


The ameliorative potential of Hyphaene thebaica on streptozotocin-induced diabetic nephropathy.
Diabetic nephropathy (DN) is the leading cause of end-stage renal disease. The aim of the current study is to investigate the possible beneficial effects of Hyphaene thebaica in DN. For this, 50 male albino rats were divided into five groups: group I - represented the control group; group II - received Hyp-haene thebaica extracts of 150 mg/kg BW by oral gavage for 6 weeks; group III - received single intraperitoneal injections of streptozotocin (50 mg/kg BW) to induce type-2 diabetes mellitus; group IV (protective) - diabetic rats recei-ved Hyphaene thebaica extract (150 mg/kg BW) orally for 6 weeks; group V (curative) - received Hyphaene thebaica extract (150 mg/kg BW) orally after the diagnosis of DN. In the DN protected group, blood glucose, urea, and creatinine decreased significantly, while insulin and C-peptide increased significantly. Moreover, cystatin C and neutrophil gelatinase-associated lipocalin decreased. Collagen fibre deposition is increased with an apparent thickening of the parietal layer of Bowman's capsules and the basal lamina of convoluted tubules, as well as increase of the immune-reaction of caspase-3 and desmin. The introduction of Hyphaene thebaica led to greater amelioration in the biochemical markers, apoptotic alterations, and podocyte injuries of the protected group than in the curative group. Hyphaene thebaica may be advised as a good choice that can delay diabetic renal complications.


Structure-Activity Studies of Bis-O-Arylglycolamides: Inhibitors of the Integrated Stress Response.
The integrated stress response comprises multiple signaling pathways for detecting and responding to cellular stress that converge at a single event-the phosphorylation of Ser51 on the α-subunit of eukaryotic translation initiation factor 2 (eIF2α). Phosphorylation of eIF2α (eIF2α-P) results in attenuation of global protein synthesis via the inhibitory effects of eIF2α-P on eIF2B, the guanine exchange factor (GEF) for eIF2. Herein we describe structure-activity relationship (SAR) studies of bis-O-arylglycolamides, first-in-class integrated stress response inhibitors (ISRIB). ISRIB analogues make cells insensitive to the effects of eIF2α-P by activating the GEF activity of eIF2B and allowing global protein synthesis to proceed with residual unphosphorylated eIF2α. The SAR studies described herein support the proposed pharmacology of ISRIB analogues as binding across a symmetrical protein-protein interface formed between protein subunits of the dimeric eIF2B heteropentamer.


Callers' experiences of making emergency calls at the onset of acute stroke: a qualitative study.
Rapid access to emergency medical services (EMS) is essential at the onset of acute stroke, but significant delays in contacting EMS often occur. To explore factors that influence the caller's decision to contact EMS at the onset of stroke, and the caller's experiences of the call. Participants were identified through a purposive sample of admissions to two hospitals via ambulance with suspected stroke. Participants were interviewed using open-ended questions and content analysis was undertaken. 50 participants were recruited (median age 62 years, 68% female). Only one of the callers (2%) was the patient. Two themes were identified that influenced the initial decision to contact EMS at the onset of stroke: perceived seriousness, and receipt of lay or professional advice. Two themes were identified in relation to the communication between the caller and the call handler: symptom description by the caller, and emotional response to onset of stroke symptoms. Many callers seek lay or professional advice prior to contacting EMS and some believe that the onset of acute stroke symptoms does not warrant an immediate 999 call. More public education is needed to improve awareness of stroke and the need for an urgent response.


Disclosing neuroimaging incidental findings: a qualitative thematic analysis of health literacy challenges.
Returning neuroimaging incidental findings (IF) may create a challenge to research participants' health literacy skills as they must interpret and make appropriate healthcare decisions based on complex radiology jargon. Disclosing IF can therefore present difficulties for participants, research institutions and the healthcare system. The purpose of this study was to identify the extent of the health literacy challenges encountered when returning neuroimaging IF. We report on findings from a retrospective survey and focus group sessions with major stakeholders involved in disclosing IF. We surveyed participants who had received a radiology report from a research study and conducted focus groups with participants, parents of child participants, Institutional Review Board (IRB) members, investigators and physicians. Qualitative thematic analyses were conducted using standard group-coding procedures and descriptive summaries of health literacy scores and radiology report outcomes are examined. Although participants reported high health literacy skills (m = 87.3 on a scale of 1-100), 67 % did not seek medical care when recommended to do so; and many participants in the focus groups disclosed they could not understand the findings described in their report. Despite their lack of understanding, participants desire to have information about their radiology results, and the investigators feel ethically inclined to return findings. The language in clinically useful radiology reports can create a challenge for participants' health literacy skills and has the potential to negatively impact the healthcare system and investigators conducting imaging research. Radiology reports need accompanying resources that explain findings in lay language, which can help reduce the challenge caused by the need to communicate incidental findings.


Oleic Acid, deglycosylated vitamin D-binding protein, nitric oxide: a molecular triad made lethal to cancer.
Oleic Acid (OA) has been shown to have anticancer properties mediated by interaction with proteins such as α-lactalbumin and lactoferrins. Therefore, we synthesized complexes of OA and Gc protein-derived macrophage activating factor (GcMAF) that inhibits per se cancer cell proliferation and metastatic potential. We hypothesised that OA-GcMAF complexes could exploit the anticancer properties of both OA and GcMAF in a synergistic manner. We postulated that the stimulating effects of GcMAF on macrophages might lead to release of nitric oxide (NO). Patients with advanced cancer were treated at the Immuno Biotech Treatment Centre with OA-GcMAF-based integrative immunotherapy in combination with a low-carbohydrate, high-protein diet, fermented milk products containing naturally-produced GcMAF, Vitamin D3, omega-3 fatty acids and low-dose acetylsalicylic acid. Measuring the tumour by ultrasonographic techniques, we observed a decrease of tumour volume of about 25%. These observations demonstrate that OA, GcMAF and NO can be properly combined and specifically delivered to advanced cancer patients with significant effects on immune system stimulation and tumour volume reduction avoiding harmful side-effects.


A Gaussian attractor network for memory and recognition with experience-dependent learning.
Attractor networks are widely believed to underlie the memory systems of animals across different species. Existing models have succeeded in qualitatively modeling properties of attractor dynamics, but their computational abilities often suffer from poor representations for realistic complex patterns, spurious attractors, low storage capacity, and difficulty in identifying attractive fields of attractors. We propose a simple two-layer architecture, gaussian attractor network, which has no spurious attractors if patterns to be stored are uncorrelated and can store as many patterns as the number of neurons in the output layer. Meanwhile the attractive fields can be precisely quantified and manipulated. Equipped with experience-dependent unsupervised learning strategies, the network can exhibit both discrete and continuous attractor dynamics. A testable prediction based on numerical simulations is that there exist neurons in the brain that can discriminate two similar stimuli at first but cannot after extensive exposure to physically intermediate stimuli. Inspired by this network, we found that adding some local feedbacks to a well-known hierarchical visual recognition model, HMAX, can enable the model to reproduce some recent experimental results related to high-level visual perception.


Improvement in the clinical practicability of roentgen stereophotogrammetric analysis (RSA): free from the use of the dual X-ray equipment.
After total knee replacement, the monitoring of the prosthetic performance is often done by roentgenographic examination. However, the two-dimensional (2D) roentgen images only provide information about the projection onto the anteroposterior (AP) and mediolateral (ML) planes. Historically, the model-based roentgen stereophotogrammetric analysis (RSA) technique has been developed to predict the spatial relationship between prostheses by iteratively comparing the projective data for the prosthetic models and the roentgen images. During examination, the prosthetic poses should be stationary. This should be ensured, either by the use of dual synchronized X-ray equipment or by the use of a specific posture. In practice, these methods are uncommon or technically inconvenient during follow-up examination. This study aims to develop a rotation platform to improve the clinical applicability of the model-based RSA technique. The rotation platform allows the patient to assume a weight-bearing posture, while being steadily rotated so that both AP and ML knee images can be obtained. This study uses X-ray equipment with a single source and flat panel detectors (FPDs). Four tests are conducted to evaluate the quality of the FPD images, steadiness of the rotation platform, and accuracy of the RSA results. The results show that the distortion-induced error of the FPD image is quite minor, and the prosthetic size can be cautiously calibrated by means of the scale ball(s). The rotation platform should be placed closer to the FPD and orthogonal to the projection axis of the X-ray source. Image overlap of the prostheses can be avoided by adjusting both X-ray source and knee posture. The device-induced problems associated with the rotation platform include the steadiness of the platform operation and the balance of the rotated subject. Sawbone tests demonstrate that the outline error, due to the platform, is of the order of the image resolution (= 0.145 mm). In conclusion, the rotation platform with steady rotation, a knee support, and a handle can serve as an alternative method to take prosthetic images, without the loss in accuracy associated with the RSA method.


Erythropoietin but not VEGF has a protective effect on auditory hair cells in the inner ear.
It has recently been shown that the oxygen-regulated factors erythropoietin (Epo) and vascular endothelial growth factor (VEGF) confer protection on different cells, including neuronal-derived ones. The receptors for Epo and VEGF are widely expressed in different organs. Since mammalian auditory hair cells can irreversibly be damaged by different agents, we aimed to identify otoprotective compounds. We focused on the role of Epo and VEGF in the inner ear and review the recent studies. Epo and its receptor are expressed in the inner ear. In vitro experiments on auditory hair cells showed a protective effect of Epo in ischemia- and gentamicin-induced hair cell damage. In contrast, an in vivo study using an animal model of noise-induced hearing loss showed a negative effect of Epo. Also VEGF and its receptors are expressed in the inner ear. Changes in the expression of VEGF or its receptors have been found in the cochlea after noise exposure, transcranial vibration and diabetic or aged animals. Until now, there are no studies about a direct effect of VEGF on auditory hair cells in vitro or in vivo. We could exclude a protective effect of VEGF on gentamicin-induced auditory hair cell damage in vitro. Thus, we conclude that Epo but not VEGF has a protective effect on auditory hair cell damage at least in vitro.


Systems-level characterization and engineering of oxidative stress tolerance in Escherichia coli under anaerobic conditions.
Despite many prior studies on microbial response to oxidative stress, our understanding of microbial tolerance against oxidative stress is currently limited to aerobic conditions, and few engineering strategies have been devised to resolve toxicity issues of oxidative stress under anaerobic conditions. Since biological processes, such as anaerobic fermentation, are frequently hampered by toxicity arising from oxidative stress, increased microbial tolerance against oxidative stress improves the overall productivity and yield of biological processes. Here, we show a systems-level analysis of oxidative stress response of Escherichia coli under anaerobic conditions, and present an engineering strategy to improve oxidative stress tolerance. First, we identified essential cellular mechanisms and regulatory factors underlying oxidative stress response under anaerobic conditions using a transcriptome analysis. In particular, we showed that nitrogen metabolisms and respiratory pathways were differentially regulated in response to oxidative stress under anaerobic and aerobic conditions. Further, we demonstrated that among transcription factors with oxidative stress-derived perturbed activity, the deletion of arcA and arcB significantly improved oxidative stress tolerance under aerobic and anaerobic conditions, respectively, whereas fnr was identified as an essential transcription factor for oxidative stress tolerance under anaerobic conditions. Moreover, we showed that oxidative stress increased the intracellular NADH : NAD(+) ratio under aerobic and anaerobic conditions, which indicates a regulatory role of NADH in oxidative stress tolerance. Based on this finding, we demonstrated that increased NADH availability through fdh1 overexpression significantly improved oxidative stress tolerance under aerobic conditions. Our results here provide novel insight into better understanding of cellular mechanisms underlying oxidative stress tolerance under anaerobic conditions, and into developing strain engineering strategies to enhance microbial tolerance against oxidative stress towards improved biological processes.


In vitro response of primary human bone marrow stromal cells to recombinant human bone morphogenic protein-2 in the early and late stages of osteoblast differentiation.
A number of factors must be added to human bone marrow stromal cells (hBMSCs) in vitro to induce osteogenesis, including ascorbic acid (AA), beta-glycerophosphate (GP), and dexamethasone (Dex). Bone morphogenic protein (BMP)-2 is an osteoinductive factor that can commit stromal cells to differentiate into osteoblasts. However, it is still not clear whether the addition of BMP-2 alone in vitro can induce hBMSCs to complete osteoblast differentiation, resulting in matrix mineralization. This study compares the effects of BMP-2 and Dex, alone and combined, on the early and late stages of hBMSC differentiation. We found that BMP-2 causes a significant induction of alkaline phosphatase (ALP) activity in hBMSCs, with a transcriptional upregulation of known BMP-2-responsive genes, and the stable expression of cbfa1 in the nucleus and the regions surrounding the nucleus in the early phase of osteoblast differentiation. However, continuous treatment with BMP-2 alone at doses ranging from 100 to 300 ng/mL results in a less efficient enhancement of in vitro matrix mineralization, despite a significant induction of ALP activity at a concentration of 100 ng/mL. Our results reflect how the effects of BMP-2 on hBMSCs can vary depending on the stage of osteoblast differentiation, and highlight the need to understand the role of BMP-2 in primary hBMSCs derived from diverse sources in order to increase the efficiency of using BMP-2 in osteoinductive therapies.


The bop2-1 mutation reveals radial asymmetry in the inner dynein arm region of Chlamydomonas reinhardtii.
Strains of Chlamydomonas reinhardtii with a mutant allele at the BOP2 locus swim slowly and have an abnormal flagellar waveform similar to previously identified strains with defects in the inner arm region. Double mutant strains with the bop2-1 allele and any of 17 different mutations that affect the dynein arm region swim more slowly than either parent, which suggests that the bop2-1 mutation does not affect solely the outer dynein arms, the I1 or ida4 inner dynein arms, or the dynein regulatory complex. Flagellar axonemes isolated from bop2-1 cells are missing a phosphorylated polypeptide of 152 kD. Electron microscopic analysis shows that bop2-1 axonemes are missing density in the inner dynein arm region. Surprisingly, two populations of images were observed in longitudinal sections of axonemes from the bop2-1 strain. In the 10 longitudinal axonemes examined, a portion of the dynein regulatory complex and a newly identified structure, the projection, are affected. In five of these 10 longitudinal axonemes examined, two lobes of the ida4 inner arm are also missing. By examining the cross-sectional images of wild-type and bop2-1 axonemes at each outer doublet position around the axoneme, we have determined that the bop2-1 mutation affects the assembly of inner arm region components in a doublet specific manner. Doublets 5, 6, and 8 have the most severe deficiency, doublet 9 has an intermediate phenotype, and doublets 2, 3, 4, and 7 have the least severe phenotype. The bop2-1 mutation provides the first evidence of radial asymmetry in the inner dynein arm region.


Dissolved organic carbon influences microbial community composition and diversity in managed aquifer recharge systems.
This study explores microbial community structure in managed aquifer recharge (MAR) systems across both laboratory and field scales. Two field sites, the Taif River (Taif, Saudi Arabia) and South Platte River (Colorado), were selected as geographically distinct MAR systems. Samples derived from unsaturated riverbed, saturated-shallow-infiltration (depth, 1 to 2 cm), and intermediate-infiltration (depth, 10 to 50 cm) zones were collected. Complementary laboratory-scale sediment columns representing low (0.6 mg/liter) and moderate (5 mg/liter) dissolved organic carbon (DOC) concentrations were used to further query the influence of DOC and depth on microbial assemblages. Microbial density was positively correlated with the DOC concentration, while diversity was negatively correlated at both the laboratory and field scales. Microbial communities derived from analogous sampling zones in each river were not phylogenetically significantly different on phylum, class, genus, and species levels, as determined by 16S rRNA gene pyrosequencing, suggesting that geography and season exerted less sway than aqueous geochemical properties. When field-scale communities derived from the Taif and South Platte River sediments were grouped together, principal coordinate analysis revealed distinct clusters with regard to the three sample zones (unsaturated, shallow, and intermediate saturated) and, further, with respect to DOC concentration. An analogous trend as a function of depth and corresponding DOC loss was observed in column studies. Canonical correspondence analysis suggests that microbial classes Betaproteobacteria and Gammaproteobacteria are positively correlated with DOC concentration. Our combined analyses at both the laboratory and field scales suggest that DOC may exert a strong influence on microbial community composition and diversity in MAR saturated zones.


Bone remodeling and bone matrix quality before and after menopause in healthy women.
Acceleration of remodeling activity after menopause leads to bone loss and fragility, however, whether this is associated with modifications of bone matrix quality has been less studied. The impact of variation in bone remodeling rate on bone matrix has been studied mainly in pathologies or anti-osteoporotic treatments. However, in healthy women this has been less studied. We analyzed, at the global level, bone matrix quality in bone biopsies from 3 groups of healthy women (20 per group): 1) before menopause (PreM), 2) 1 year after menopause (PostM, paired biopsies with preM), and 3) 14 (±9) years after menopause (LT-PostM). The mean degree of mineralization (DMB) and heterogeneity index (HI) of mineralization were assessed by X-ray microradiography on whole bone matrix; intrinsic properties (mineral/organic ratio, mineral maturity, mineral crystallinity, collagen maturity) were assessed by Fourier Transform Infrared microspectroscopy, microhardness by microindentation, both at a global level and calculated by mean of several measurements over the whole tissue area. In PostM compared to PreM (bone remodeling rate had doubled), mean DMB measured on the entire bone plane (whole bone matrix) of the sample was not different. HI was increased in trabecular bone indicating a higher heterogeneity of mineralization. However, in PostM, mineral/organic ratio (trabecular) and microhardness (cortical and trabecular) were decreased, whereas mineral/collagen maturation or crystal size/perfection were unchanged. Thus, in PostM, the local mineral content and microhardness were first affected. In LT-PostM (bone remodeling rate was 3 times higher), the mean DMB was still not different. However, the mineral/organic ratio, microhardness, mineral maturity, crystallinity all were lower compared to PreM and PostM, in both cortical and trabecular bone. Bone remodeling rate was negatively correlated with microhardness, DMB, mineral/organic and crystallinity. This suggests that increases in bone remodeling rates after menopause have a direct impact on bone quality by inducing the formation of more extensive "immature" bone areas, but the amount of immature bone does not cause modification of the global DMB.


Asymmetry in a pterobranch hemichordate and the evolution of left-right patterning.
To investigate the possible origin of vertebrate asymmetry, we reexamined the long-described asymmetry of pterobranch hemichordates, a group of tiny, mostly colonial marine filter feeders. As first described over a century ago, we found left-right (LR) asymmetry in localization of the gonad. However, contrary to early descriptions, the direction of asymmetry is random, displaying antisymmetry rather than directional asymmetry. We found that the direction of gonad localization is significantly related to the shape of the tube in which the zooid lives. We also reexamined other anatomical features described to have directional asymmetry, the shape of the oral lamella and localization of the gonopore; however, we did not confirm asymmetry in these examples. Together with previous studies, our results suggest that hemichordates have undergone degeneration in the precision of asymmetry from an ancestral asymmetrical condition that existed before the divergence of deuterostomes.


Differential effects of anti-Nogo-A antibody treatment and treadmill training in rats with incomplete spinal cord injury.
Locomotor training on treadmills can improve recovery of stepping in spinal cord injured animals and patients. Likewise, lesioned rats treated with antibodies against the myelin associated neurite growth inhibitory protein, Nogo-A, showed increased regeneration, neuronal reorganization and behavioural improvements. A detailed kinematic analysis showed that the hindlimb kinematic patterns that developed in anti-Nogo-A antibody treated versus treadmill trained spinal cord injured rats were significantly different. The synchronous combined treatment group did not show synergistic effects. This lack of synergistic effects could not be explained by an increase in pain perception, sprouting of calcitonin gene-related peptide (CGRP) positive fibres or by interference of locomotor training with anti-Nogo-A antibody induced regeneration and sprouting of descending fibre tracts. The differential mechanisms leading to behavioural recovery during task-specific training and in regeneration or plasticity enhancing therapies have to be taken into account in designing combinatorial therapies so that their potential positive interactive effects can be fully expressed.


Ifosfamide neurotoxicty in a young female with a remarkable response to thiamine.
Ifosfamide, a commonly used chemotherapeutic agent in various regimens for many malignancies and has a well known central nervous system side effect. Ifosfamide induced encephalopathy develops in approximately 10-30% of patients exposed to the drug. It is generally reversible after discontinuing the therapy; however cases of fatal neurotoxicity have been reported in literature. Commonly used antidote, Methylene blue; has a moderate efficacy in reversing the encephalopathy followed by lesser response rates by Thiamine. We submit a case report of a young female patient with refractory diffuse large B cell lymphoma who developed severe ifosfamide neurotoxicity. With the use of intravenous thiamine, encephalopathy resolved in our patient within a mean time of 30 hours (average range is 10-30 hours). We found Thiamine to be safe and effective in treatment for ifosfamide induced encephalopathy.


Hierarchical regulation of the NikR-mediated nickel response in Helicobacter pylori.
Nickel is an essential metal for Helicobacter pylori, as it is the co-factor of two enzymes crucial for colonization, urease and hydrogenase. Nickel is taken up by specific transporters and its intracellular homeostasis depends on nickel-binding proteins to avoid toxicity. Nickel trafficking is controlled by the Ni(II)-dependent transcriptional regulator NikR. In contrast to other NikR proteins, NikR from H. pylori is a pleiotropic regulator that depending on the target gene acts as an activator or a repressor. We systematically quantified the in vivo Ni(2+)-NikR response of 11 direct NikR targets that encode functions related to nickel metabolism, four activated and seven repressed genes. Among these, four targets were characterized for the first time (hpn, hpn-like, hydA and hspA) and NikR binding to their promoter regions was demonstrated by electrophoretic mobility shift assays. We found that NikR-dependent repression was generally set up at higher nickel concentrations than activation. Kinetics of the regulation revealed a gradual and temporal NikR-mediated response to nickel where activation of nickel-protection mechanisms takes place before repression of nickel uptake. Our in vivo study demonstrates, for the first time, a chronological hierarchy in the NikR-dependent transcriptional response to nickel that is coherent with the control of nickel homeostasis in H. pylori.


Analysis of the carbon-13 and proton NMR spectra of bovine chromaffin granules.
Natural abundance carbon-13 and proton NMR spectra of bovine chromaffin granules have been obtained and analyzed using computer simulation techniques. High resolution spectra show the presence of a fluid aqueous phase containing epinephrine, ATP and a random coil protein. The protein spectrum contains unusually intense resonances due to glutamic acid and proline and has been simulated satisfactorily using the known amino acid composition of chromogranin A. The lipid phase of chromaffin granules gives rise to intense, but very broad, resonances in the carbon-13 spectrum. Protons in the lipid phase are also observable as a very rapid component of the proton-free induction decay (T2 approximately equal to 15 microns). Linewidths of the carbon-13 spectra have been used to set upper limits on rotational correlation times and on the motional anisotropy in the aqueous phase. These limits show that the aqueous phase is a simple solution (not a gel) that is isotropic over regions much larger than solute dimensions. No gel transition is observed between -3 and 25 degrees C. The carbon-13 spectra are definitely inconsistent with a lipoprotein matrix model and chromaffin granules previously proposed by Helle and Serck-Hanssen ((1975) Mol. Cell, Biochem. 6, 127-146). Relative carbon-13 intensities of ATP and epinephrine are not consistent with the known 1 : 4 mol ratio of these components. This fact suggests that epinephrine and ATP are not directly complexed in intact chromaffin granules.


Evaluation of the role of miR-31-dependent reduction in dystrophin and nNOS on atrial-fibrillation-induced electrical remodelling in man.
The management of atrial fibrillation remains a challenge. This condition remodels atrial electrical properties, which promote resistance to treatment. Although remodelling has long been a therapeutic target in atrial fibrillation, its causes remain incompletely understood. We aimed to evaluate the role of miR-31-dependent reduction in dystrophin and neuronal nitric oxide synthase (nNOS, also known as NOS1) on atrial electrical properties and atrial fibrillation inducibility. We recruited 258 patients (209 patients in sinus rhythm and 49 with permanent atrial fibrillation) from the John Radcliffe Hospital, Oxford, UK; written informed consent was obtained from each participant. We also used a goat model of pacing-induced atrial fibrillation (24 with atrial fibrillation vs 20 controls in normal sinus rythm) and nNos-knock-out mice (n=28 compared with 27 wild-type littermates). Gene expression of miR-31, dystrophin, and nNOS was assessed by quantitative RT-PCR; protein content was measured by immunoblotting; NOS activity was evaluated with high-performance liquid chromatography; action potential duration (APD) and rate dependent adaptation were assessed by single-cell patch-clamping, and atrial fibrillation inducibility was evaluated by transoesophageal atrial burst stimulation. We found that atrial-specific upregulation of miR-31 in human atrial fibrillation caused dystrophin (DYS) translational repression and accelerated mRNA degradation of nNOS leading to a profound reduction in atrial DYS and nNOS protein content and in nitric oxide availability. In human atrial myocytes obtained from patients in sinus rhythm, nNOS inhibition was sufficient to recapitulate hallmark features of remodelling induced by atrial fibrillation, such as shortening of APD and loss of APD rate-dependency, but had no effect in patients with atrial fibrillation. In mice, nNos gene deletion or inhibition shortened atrial APD and increased atrial fibrillation inducibility in vivo. Inhibition of miR-31 in human atrial fibrillation recovered DYS and nNOS, and normalised APD and APD rate-dependency. Prevention of miR-31 binding to nNOS 3'UTR recovered both nNOS protein and gene expression but had no effect on the DYS protein or mRNA level (consistent with the mRNA degradation of nNOS by miR-31). Prevention of miR-31 binding to DYS 3'UTR increased DYS protein but not mRNA is consistent with translation repression of DYS by miR-31; recovery of DYS protein increased nNOS protein but not mRNA in keeping with a stabilising effect of DYS on nNOS protein. In goats, a reduction in dystrophin and nNOS protein content was associated with upregulation of miR-31 in the atria but not in the ventricles. The findings suggest that atrial-specific upregulation of miR-31 in human atrial fibrillation is a key mechanism causing atrial loss of dystrophin and nNOS; this loss leads to the electrical phenotype induced by atrial fibrillation. British Heart Foundation (BHF) Programme grant (for BC and XL), BHF Centre of Excellence in Oxford (SR), Leducq Foundation (in part for BC and SR), the European Union's seventh Framework Programme Grant Agree.


Effect of cariogenic challenge on the stability of dentin bonds.
The oral environment is subject to biofilm accumulation and cariogenic challenge, and few studies exist on the effect of these factors on the bond strength of adhesive systems. The aim of this study was to test if the exposure of adhesive interfaces to cariogenic challenge under biofilm accumulation could promote higher degradation than the exposure to biofilm accumulation alone. Five molars were ground until exposure of medium dentin and then restored (Single Bond 2 and Z250 3M ESPE). The tooth/resin sets were cut to obtain beam-shaped specimens, which were distributed according to the aging conditions (n=20): water for 24 h (control); biofilm under cariogenic challenge for 3, 5 or 10 days; biofilm without cariogenic challenge for 10 days; and water for 3 months. Microcosm biofilms were formed from human saliva and grown in a saliva analogue medium, supplemented or not with sucrose to promote cariogenic challenge. Specimens were tested for microtensile bond strength, and failure modes were classified using light microscopy. Bond strength data were analyzed using ANOVA and failure modes were analyzed using ANOVA on ranks (α=0.05). No significant differences in bond strength were detected among the aging methods (P=0.248). The aging period was associated with an increase in the frequency of adhesive failures for the groups aged for 10 days or longer (P<0.001). Aging leads to a higher prevalence of interfacial adhesive failures, although this effect is not associated with cariogenic challenge or reduction in bond strengths.


Policy statement--Role of the pediatrician in youth violence prevention.
Youth violence continues to be a serious threat to the health of children and adolescents in the United States. It is crucial that pediatricians clearly define their role and develop the appropriate skills to address this threat effectively. From a clinical perspective, pediatricians should become familiar with Connected Kids: Safe, Strong, Secure, the American Academy of Pediatrics' primary care violence prevention protocol. Using this material, practices can incorporate preventive education, screening for risk, and linkages to community-based counseling and treatment resources. As advocates, pediatricians may bring newly developed information regarding key risk factors such as exposure to firearms, teen dating violence, and bullying to the attention of local and national policy makers. This policy statement refines the developing role of pediatricians in youth violence prevention and emphasizes the importance of this issue in the strategic agenda of the American Academy of Pediatrics.


Visual Analytics for Electromagnetic Situation Awareness in Radio Monitoring and Management.
Traditional radio monitoring and management largely depend on radio spectrum data analysis, which requires considerable domain experience and heavy cognition effort and frequently results in incorrect signal judgment and incomprehensive situation awareness. Faced with increasingly complicated electromagnetic environments, radio supervisors urgently need additional data sources and advanced analytical technologies to enhance their situation awareness ability. This paper introduces a visual analytics approach for electromagnetic situation awareness. Guided by a detailed scenario and requirement analysis, we first propose a signal clustering method to process radio signal data and a situation assessment model to obtain qualitative and quantitative descriptions of the electromagnetic situations. We then design a two-module interface with a set of visualization views and interactions to help radio supervisors perceive and understand the electromagnetic situations by a joint analysis of radio signal data and radio spectrum data. Evaluations on real-world data sets and an interview with actual users demonstrate the effectiveness of our prototype system. Finally, we discuss the limitations of the proposed approach and provide future work directions.


Life-history traits drive the evolutionary rates of mammalian coding and noncoding genomic elements.
A comprehensive phylogenetic framework is indispensable for investigating the evolution of genomic features in mammals as a whole, and particularly in humans. Using the ENCODE sequence data, we estimated mammalian neutral evolutionary rates and selective pressures acting on conserved coding and noncoding elements. We show that neutral evolutionary rates can be explained by the generation time (GT) hypothesis. Accordingly, primates (especially humans), having longer GTs than other mammals, display slower rates of neutral evolution. The evolution of constrained elements, particularly of nonsynonymous sites, is in agreement with the expectations of the nearly neutral theory of molecular evolution. We show that rates of nonsynonymous substitutions (dN) depend on the population size of a species. The results are robust to the exclusion of hypermutable CpG prone sites. The average rate of evolution in conserved noncoding sequences (CNCs) is 1.7 times higher than in nonsynonymous sites. Despite this, CNCs evolve at similar or even lower rates than nonsynonymous sites in the majority of basal branches of the eutherian tree. This observation could be the result of an overall gradual or, alternatively, lineage-specific relaxation of CNCs. The latter hypothesis was supported by the finding that 3 of the 20 longest CNCs displayed significant relaxation of individual branches. This observation may explain why the evolution of CNCs fits the expectations of the nearly neutral theory less well than the evolution of nonsynonymous sites.


Channel-mediated potassium uptake in Helicobacter pylori is essential for gastric colonization.
To date, the biological role of prokaryotic K(+) channels remains unknown. Helicobacter pylori contains a gene encoding a putative K(+) channel (HpKchA) of the two-transmembrane RCK (regulation of K(+) conductance) domain family, but lacks known bacterial K(+) uptake systems. A H. pylori DeltahpKchA mutant presented a strong growth defect at low K(+) concentration, which was compensated by KCl addition. The role of the separate RCK domain was investigated in H. pylori by mutagenesis of its internal start codon, which led to a K(+)-dependent intermediate growth phenotype, consistent with RCK activating channel function. Tagging HpKchA C-terminally, we detected a 1:1 stoichiometry of the full-length HpKchA and the separate RCK domain. We constructed single amino-acid exchanges within the unusual selectivity filter of HpKchA (ATGFGA) in H. pylori and observed complete loss (G74A), a slight defect (G76A or F75G) or wild-type (A77D) channel function. HpKchA was essential for colonization of the murine stomach. These data show, for the first time, a biological function for a prokaryotic K(+) channel, as a K(+) uptake system, essential for the persistence of H. pylori in the gastric environment.


Characterisation of Chlamydia pneumoniae and other novel chlamydial infections in captive snakes.
Chlamydiosis has been described in both free-ranging and captive reptiles. The infection usually manifests as granulomatous inflammation in inner organs such as spleen, heart, lung and liver but might also occur in asymptomatic reptiles. The aim of this study was to investigate and characterise Chlamydia pneumoniae and potential other novel chlamydial infections in the choana and cloaca samples of 137 clinically healthy captive snakes from six private collections. Forty eight samples from 29 animals were found to be positive by a Chlamydiaceae family-specific qPCR. By Chlamydia species-specific ArrayTube Microarray, 43 samples were positive, with 36 of these being identified as C. pneumoniae. The prevalence of Chlamydia ranged from 5 to 33%. PCR and sequencing of the Chlamydiales 16S rRNA signature sequence of 21 Chlamydia positive samples revealed the presence of seven novel 16S rRNA genotypes. BLAST-n and phylogenetic analysis of the near-full length 16S rRNA gene sequence of each of these novel 16S rRNA sequences revealed that five genotypes share closest sequence identity to 16S rRNA sequences from C. pneumoniae (98.6-99.2%), suggesting that these sequences are novel C. pneumoniae strains. One genotype is 96.9% similar to C. pneumoniae strains suggesting it may originate from a yet undescribed chlamydial species within the genus Chlamydia. This study further highlights the broad host range for C. pneumoniae and suggests that reptiles may still contain a significant and largely uncharacterised level of chlamydial genetic diversity that requires further investigation.


Residue arithmetic processing utilizing optical Fredkin gate arrays.
A cascadable residue arithmetic processor based on optical Fredkin gate arrays and page-oriented holographic memories is introduced. The implementations of residue functions and operations by this processor are described. Analytic expressions are derived for the number of holograms and waveguide channels required for the implementation of residue addition and multiplication. The practical cases of 16-bit addition and multiplication are analyzed as specific examples. It is shown that, using the proposed architecture, these operations can be implemented with state-of-the-art technologies in holography and integrated optics.


Characterization of the calcium-mediated response to alkaline stress in Saccharomyces cerevisiae.
Exposure of the yeast Saccharomyces cerevisiae to alkaline stress resulted in adaptive changes that involved remodeling the gene expression. Recent evidence suggested that the calcium-activated protein phosphatase calcineurin could play a role in alkaline stress signaling. By using an aequorin luminescence reporter, we showed that alkaline stress resulted in a sharp and transient rise in cytoplasmic calcium. This increase was largely abolished by addition of EGTA to the medium or in cells lacking Mid1 or Cch1, components of the high affinity cell membrane calcium channel. Under these circumstances, the alkaline response of different calcineurin-sensitive transcriptional promoters was also blocked. Therefore, exposure to alkali resulted in entry of calcium from the external medium, and this triggered a calcineurin-mediated response. The involvement of calcineurin and Crz1/Tcn1, the transcription factor activated by the phosphatase, in the transcriptional response triggered by alkalinization has been globally assessed by DNA microarray analysis in a time course experiment using calcineurin-deficient (cnb1) and crz1 mutants. We found that exposure to pH 8.0 increased at least 2-fold the mRNA levels of 266 genes. In many cases (60%) the response was rather early (peak after 10 min). The transcriptional response of 27 induced genes (10%) was reduced or fully abolished in cnb1 cells. In general, the response of crz1 mutants was similar to that of calcineurin-deficient cells. By analysis of a systematic deletion library, we found 48 genes whose mutation resulted in increased sensitivity to the calcineurin inhibitor FK506. Twenty of these mutations (42%) also provoked alkaline pH sensitivity. In conclusion, our results demonstrated that calcium signaling and calcineurin activation represented a significant component of the yeast response to environmental alkalinization.


A case of primary amebic meningoencephalitis. Light and electron microscopy, and immunohistologic studies.
A 56-year-old alcoholic developed primary meningoencephalitis and died 3 weeks later. Autopsy revealed multiple areas of necrosis and acute and granulomatous inflammation in the brain and pancreas. Trophozoites and cysts were discovered in the brain and trophozoites alone in the pancreas. Morphologic studies revealed some parasitic features described as characteristic for Naegleria, but clinical, histological and immunohistological features favored in Acanthamoeba infection. This study calls for caution in the use of pure morphological criteria for the differential diagnosis of Naegleria and Acanthamoeba in humans. Cultural characteristics, immunohistology and serologic tests are probably more reliable for differentiating these parasites.


Tissue-specific expression of the heat shock protein HSP27 during Drosophila melanogaster development.
The alpha-crystallin-related heat shock (stress) protein hsp27 is expressed in absence of heat shock during Drosophila melanogaster development. Here, we describe the tissue distribution of this protein using an immunoaffinity-purified antibody. In embryos, hsp27 translated from maternal RNA is uniformly distributed, except in the yolk. During the first, second, and early third larval stages, hsp27 expression is restricted to the brain and the gonads. These tissues are characterized by a high level of proliferating cells. In late third instar larvae and early pupae, in addition to the central nervous system and the gonads, all the imaginal discs synthesize hsp27. The disc expression seems restricted to the beginning of their differentiation since it disappears during the second half of the pupal stage: no more hsp27 is observed in the disc-derived adult organs. In adults, hsp27 is still present in some regions of the central nervous system, and is also expressed in the male and female germ lines where it accumulates in mature sperm and oocytes. The transcript and the protein accumulate in oocytes since the onset of vitellogenesis with a uniform distribution similar to that found in embryos. The adult germ lines transcribe hsp27 gene while no transcript is detected in the late pupal and adult brain. These results suggest multiple roles of hsp27 during Drosophila development which may be related to both the proliferative and differentiated states of the tissues.


Effects of interoceptive accuracy in autonomic responses to external stimuli based on cardiac rhythm.
Interoceptive accuracy is an index of the ability to perceive an individual's internal bodily state, including heartbeat and respiration. Individual differences in interoceptive accuracy influence emotional recognition through autonomic nervous activity. However, the precise mechanism by which interoceptive accuracy affects autonomic reactivity remains unclear. Here, we investigated how cardiac reactivity induced by a non-affective external rhythm differed among individuals, using a heartbeat counting task. Because individuals with poor interoceptive accuracy cannot distinguish an external rhythm from their cardiac cycles, it has been hypothesized that the interoceptive effect on heart rate works differently in individuals with good interoceptive accuracy and those with poor interoceptive accuracy. Study participants observed a visual or auditory stimulus presented at a rhythm similar to the participants' resting heart rates. The stimulus rhythm was gradually changed from that of their resting heart rate, and we recorded electrocardiographs while participants were exposed to the stimuli. Individuals with good interoceptive accuracy exhibited a deceleration in heart rate when the rhythm of the auditory stimulus changed. In contrast, in the group with poor interoceptive accuracy, the heart rate decreased only when the stimulus became faster. They were unable to distinguish the rhythm of their own heartbeat from that of the external rhythm; therefore, we propose that such individuals recognize the stimuli at the pace of their heart rate. Individuals with good interoceptive accuracy were able to distinguish their heart rates from the external rhythm. A modality difference was not observed in this study, which suggests that both visual and auditory stimuli help mimic heart rate. These results may provide physiological evidence that autonomic reactivity influences the perception of the internal bodily state, and that interoception and the autonomic state interact to some degree.


Effectiveness of Thrombolytic Therapy in Acute Embolic Stroke due to Infective Endocarditis.
Objective. To identify the role of thrombolytic therapy in acute embolic stroke due to infective endocarditis. Design. Case report. Setting. University hospital. Patient. A 70-year-old male presented with acute onset aphasia and hemiparesis due to infective endocarditis. His head computerized tomographic scan revealed left parietal sulcal effacement. He was given intravenous tissue plasminogen activator with significant resolution of the neurologic deficits without complications. Main Outcome Measures. Physical examination, National Institute of Health Stroke Scale, radiologic examination results. Conclusions. Thrombolytic therapy in selected cases of stroke due to infective endocarditis manifesting as major neurologic deficits can be considered as an option after careful consideration of risks and benefits. The basis for such favorable response rests in the presence of fibrin as a major constituent of the vegetation. The risk of precipitating hemorrhage with thrombolytic therapy especially with large infarcts and mycotic aneurysms should be weighed against the benefits of averting a major neurologic deficit.