Genome-wide analyses of borderline personality features.
The heritability of borderline personality (BP) features has been established in multiple twin and family studies. Using data from the borderline subscale of the Personality Assessment Inventory Borderline Features Scale (PAI-BOR) collected in two Dutch cohorts (N=7125), the Netherlands Twin Register and The Netherlands Study of Depression and Anxiety, we show that heritability of the PAI-BOR total score using genome-wide single-nucleotide polymorphism (SNPs) is estimated at 23%, and that the genetic variance is substantially higher in affect instability items compared with the other three subscales of the PAI-BOR (42.7% vs non-significant estimates for self-harm, negative relations and identity problems). We present results from a first genome-wide association study of BP features, which shows a promising signal on chromosome 5 corresponding to SERINC5, a protein involved in myelination. Reduced myelination has been suggested as possibly having a role in the development of psychiatric disorders characterized by lack of social interaction. The signal was confirmed in a third independent Dutch cohort drawn from the Erasmus Rucphen Family study (N=1301). Our analyses were complemented by investigating the heterogeneity that was implied by the differences in genetic variance components in the four subscales of the PAI-BOR. These analyses show that the association of SNPs tagging SERINC5 differs substantially across the 24 items of the PAI-BOR. Further, using reverse regression we showed that the effects were present only in subjects with higher scores on the PAI-BOR. Taken together, these results suggest that future genome-wide analyses can benefit substantially by taking into account the phenotypic and genetic heterogeneity of BP features.


Postmortem memantine concentrations.
Postmortem fluid and tissue concentrations of memantine (Namenda), a drug recently approved for the treatment of Alzheimer's Disease by the FDA, are reported in a suspicious death. In addition, memantine concentrations considered to be incidental findings in three other cases are included to aid in the interpretation in future toxicological investigations. Memantine was extracted from biological samples by a standard liquid-liquid basic drug method followed by analysis utilizing a gas chromatograph-mass spectrometer operated in SIM mode. Blood concentrations ranged from 0.03 to 1.8 mg/L, and the liver concentration was 6.1 mg/kg.


Methods for the induction of reproduction in a tropical species of filamentous ulva.
The green seaweed Ulva is a major fouling organism but also an edible aquaculture product in Asia. This study quantified for the first time the effect of key factors on the reproduction of a tropical species of filamentous Ulva (Ulva sp. 3). The controlled timing of release of swarmers (motile reproductive bodies) was achieved when experiments were initiated in the early afternoon by exposing the thalli to a temperature shock (4°C) for 10 min and subsequently placing them into autoclaved filtered seawater under a 12 h light: 12 h dark photoperiod at 25°C. The release of swarmers then peaked two days after initiation. In contrast, segmentation, dehydration, salinity or time of initiation of experiments had no effect of any magnitude on reproduction. The released swarmers were predominantly biflagellate (95%), negatively phototactic and germinated without complementary gametes. This indicates that Ulva sp. 3 has a simple asexual life history dominated by biflagellate zoids.


A Review of Deep Learning for Screening, Diagnosis, and Detection of Glaucoma Progression.
Because of recent advances in computing technology and the availability of large datasets, deep learning has risen to the forefront of artificial intelligence, with performances that often equal, or sometimes even exceed, those of human subjects on a variety of tasks, especially those related to image classification and pattern recognition. As one of the medical fields that is highly dependent on ancillary imaging tests, ophthalmology has been in a prime position to witness the application of deep learning algorithms that can help analyze the vast amount of data coming from those tests. In particular, glaucoma stands as one of the conditions where application of deep learning algorithms could potentially lead to better use of the vast amount of information coming from structural and functional tests evaluating the optic nerve and macula. The purpose of this article is to critically review recent applications of deep learning models in glaucoma, discussing their advantages but also focusing on the challenges inherent to the development of such models for screening, diagnosis and detection of progression. After a brief general overview of deep learning and how it compares to traditional machine learning classifiers, we discuss issues related to the training and validation of deep learning models and how they specifically apply to glaucoma. We then discuss specific scenarios where deep learning has been proposed for use in glaucoma, such as screening with fundus photography, and diagnosis and detection of glaucoma progression with optical coherence tomography and standard automated perimetry. Deep learning algorithms have the potential to significantly improve diagnostic capabilities in glaucoma, but their application in clinical practice requires careful validation, with consideration of the target population, the reference standards used to build the models, and potential sources of bias.


An alternative-exon database and its statistical analysis.
We compiled a comprehensive database of alternative exons from the literature and analyzed them statistically. Most alternative exons are cassette exons and are expressed in more than two tissues. Of all exons whose expression was reported to be specific for a certain tissue, the majority were expressed in the brain. Whereas the length of constitutive exons follows a normal distribution, the distribution of alternative exons is skewed toward smaller ones. Furthermore, alternative-exon splice sites deviate more from the consensus: their 3' splice sites are characterized by a higher purine content in the polypyrimidine stretch, and their 5' splice sites deviate from the consensus sequence mostly at the +4 and +5 positions. Furthermore, for exons expressed in a single tissue, adenosine is more frequently used at the -3 position of the 3' splice site. In addition to the known AC-rich and purine-rich exonic sequence elements, sequence comparison using a Gibbs algorithm identified several motifs in exons surrounded by weak splice sites and in tissue-specific exons. Together, these data indicate a combinatorial effect of weak splice sites, atypical nucleotide usage at certain positions, and functional enhancers as an important contribution to alternative-exon regulation.


Should a diagnosis of endometriosis be sought in all symptomatic women?
A tool is needed to enable clinicians to determine whether women wish to seek a pathology-based explanation for chronic pelvic pain or whether they just want symptom relief. Such an approach might reduce the number of unnecessary laparoscopies without adversely affecting outcomes.


Imperatorin exerts antiallergic effects in Th2-mediated allergic asthma via induction of IL-10-producing regulatory T cells by modulating the function of dendritic cells.
Imperatorin is a furanocoumarin compound which exists in many medicinal herbs and possesses various biological activities. Herein, we investigated the antiallergic effects of imperatorin in asthmatic mice and explored the immunomodulatory actions of imperatorin on immune cells. We used a murine model of ovalbumin (OVA)-induced asthma to evaluate the therapeutic potential of imperatorin. Additionally, bone marrow-derived dendritic cells (DCs; BMDCs) were used to clarify whether imperatorin exerts an antiallergic effect through altering the ability of DCs to regulate T cells. Oral administration of imperatorin to OVA-sensitized and -challenged mice decreased serum OVA-specific immunoglobulin E (IgE) production, attenuated the airway hyperresponsiveness (AHR), and alleviated airway inflammation in a dose-dependent manner. Notably, secretions of Th2 cytokines and chemokines were reduced, and numbers of interleukin (IL)-10-producing regulatory T cells (Tregs) increased in imperatorin-treated mice. Imperatorin inhibited proinflammatory cytokines and IL-12 production but enhanced IL-10 secretion by lipopolysaccharide (LPS)-stimulated BMDCs. Compared to fully mature DCs, imperatorin-treated DCs expressed high levels of the inducible costimulatory ligand (ICOSL) and Jagged1 molecules, and had the regulatory capacity to promote the generation of IL-10-producing CD4(+) T cells in vitro. Additionally, imperatorin directly suppressed activated CD4(+) T-cell proliferation and cytokine production. Imperatorin may possess therapeutic potential against Th2-mediated allergic asthma not only via stimulating DC induction of Tregs but also via direct inhibition of Th2 cell activation. These findings provide new insights into how imperatorin affects the Th2 immune response and the development of imperatorin as a Treg-type immunomodulatory agent to treat allergic asthma.


Predictors of the risk of malnutrition among children under the age of 5 years in Somalia.
To investigate the predictors of wasting, stunting and low mid-upper arm circumference among children aged 6-59 months in Somalia using data from household cross-sectional surveys from 2007 to 2010 in order to help inform better targeting of nutritional interventions. Cross-sectional nutritional assessment surveys using structured interviews were conducted among communities in Somalia each year from 2007 to 2010. A two-stage cluster sampling methodology was used to select children aged 6-59 months from households across three livelihood zones (pastoral, agro-pastoral and riverine). Predictors of three anthropometric measures, weight-for-height (wasting), height-for-age (stunting) and mid-upper arm circumference, were analysed using Bayesian binomial regression, controlling for both spatial and temporal dependence in the data. The study was conducted in randomly sampled villages, representative of three livelihood zones in Somalia. Children between the ages of 6 and 59 months in Somalia. The estimated national prevalence of wasting, stunting and low mid-upper arm circumference in children aged 6-59 months was 21 %, 31 % and 36 %, respectively. Although fever, diarrhoea, sex and age of the child, household size and access to foods were significant predictors of malnutrition, the strongest association was observed between all three indicators of malnutrition and the enhanced vegetation index. A 1-unit increase in enhanced vegetation index was associated with a 38 %, 49 % and 59 % reduction in wasting, stunting and low mid-upper arm circumference, respectively. Infection and climatic variations are likely to be key drivers of malnutrition in Somalia. Better health data and close monitoring and forecasting of droughts may provide valuable information for nutritional intervention planning in Somalia.


Effects of serotonin on rat ileocolonic transit and fluid transfer in vivo: possible mechanisms of action.
The aim was to investigate the action of serotonin (5HT) on function of the ileocolonic junction (ICJ) in vivo. In anaesthetised rats, models were developed to study the effects of intra-aortic (ia) serotonin on ileocolonic and colonic transit, and the effects on transit of a number of 5HT receptor antagonists. In the first series of experiments, a bolus of saline labelled with 99mTc DTPA was instilled 20 cm proximal to the ICJ and transit was assessed three hours later by the geometric centre of the spread of isotope. In the second series, similar techniques were used on the postcaecal colon and transit assessed two hours later. In the third series of experiments, the effects of ia 5HT on ileal net fluid flux was evaluated by standard perfusion experiments with 14C polyethylene glycol (PEG) 4000 as a non-absorbable marker in rat plasma-like electrolyte solution. Compared with ia saline, 5HT accelerated ICJ transit significantly (p < 0.05). This acceleration was comparable with the effect of ia bethanechol. The effects of 5HT on ICJ transit were inhibited by the intraperitoneal (ip) infusion of atropine, the 5HT receptor antagonists, methysergide, ketanserin, zacopride, and the 5HT4 agonist, SC53116. Methysergide, zacopride, and SC53116 given with ia 5HT slowed ICJ transit to rates below those of ia saline alone. When these same agents were given together with ia saline, the ICJ transit was not significantly altered. Serotonin, at the dose that accelerated ICJ transit, did not significantly alter colonic transit or ileal fluid transport. In conclusion, 5HT is a potent pharmacological stimulant of transit across the rat ICJ in vivo; the action of 5HT is mediated partly through muscarinic neurones and several 5HT receptor subtypes.


American cranberries and health benefits - an evolving story of 25 years.
Cranberries contain various types of bioactive components. Scientists have been studying cranberries' beneficial effects on urinary tract health since the 20th century. In the 21st century, the protection provided by cranberry phytochemicals against cancer and vascular diseases has drawn more attention from researchers. Anthocyanins, procyanidins, and flavonols in cranberries were all documented to have potential effects on cancer prevention. The cardiometabolic effects of cranberries have been investigated in several clinical trials. It was found that cranberries positively affect atherosclerotic cholesterol profiles and that they reduced several cardiometabolic risk factors. Nowadays, growing evidence suggests other important roles of cranberries in maintaining digestive health. Cranberry juice or cranberries have been shown to inhibit the colonization of H. pylori in stomach, and protect against intestinal inflammation. For future research, clinical trials with improved study design are urgently needed to demonstrate cranberries' benefits on urinary tract health and cardiometabolic diseases. Hypothesis-driven studies using animals or cell culture are needed to elucidate the mechanisms of cranberries' effects on digestive health. © 2018 Society of Chemical Industry.


Synthesis and Biological Evaluation of Novel Hybrid Molecules Containing Purine, Coumarin and Isoxazoline or Isoxazole Moieties.
The 1,3-dipolar cycloaddition reactions of nitrile oxides formed in situ (in the presence of NCS and Et3N) from the oximes of (purin-9-yl)acetaldehyde or (coumarinyloxy)acetaldehyde with allyloxycoumarins or 9-allylpurines, respectively resulted in 3,5-disubstituted isoxazolines. The similar reactions of propargyloxycoumarins or 9-propargylpurines led to 3,5-disubstituted isoxazoles by treatment with PIDA and catalytic amount of TFA. The new compounds were tested in vitro as antioxidant agents and inhibitors of soybean lipoxygenase LO, AChE and MAO-B. The majority of the compounds showed significant hydroxyl radical scavenging activity. Compounds 4k and 4n presented LO inhibitory activity. Compound 13e presents an antioxidant significant profile combining anti-LO, anti-AChE and anti-MAO-B activities.


Comparison of thicknesses of the myocardial fibers of anencephalic and normal human fetuses.
When heart transplantation is needed in newborns, brain death should be confirmed, and the heart should not be exposed to hypoxia. The anencephalic newborn has been presented as a donor in heart transplantation. It is important, therefore, to evaluate possible morphological differences in the hearts of anencephalic cases. In this study, muscle fibers were studied in 10 anencephalic and 10 normal fetuses (27-35 weeks) and the results were compared. Random samples were taken from the upper 1/3 of the right ventricle's posterior wall and processed for light microscopic examination. Thicknesses of the 100 myocardial muscle fibers for each fetus were evaluated. There was statistically no significant difference between the anencephalic and normal fetus groups and the sex groups. Morphological features of the transplant probably affects the performance of the heart after operation. The anencephalic fetuses could be unique donors for heart transplantation.


Ovarian Cancer, Version 1.2016, NCCN Clinical Practice Guidelines in Oncology.
This selection from the NCCN Guidelines for Ovarian Cancer focuses on the less common ovarian histopathologies (LCOHs), because new algorithms were added for LCOHs and current algorithms were revised for the 2016 update. The new LCOHs algorithms include clear cell carcinomas, mucinous carcinomas, and grade 1 (low-grade) serous carcinomas/endometrioid epithelial carcinomas. The LCOHs also include carcinosarcomas (malignant mixed Müllerian tumors of the ovary), borderline epithelial tumors (also known as low malignant potential tumors), malignant sex cord-stromal tumors, and malignant germ cell tumors.


Cardiovascular burden and percutaneous interventions in Russian Federation: systematic epidemiological update.
The situation with cardiovascular (CV) burden in Russian population is alerting, and becomes of interest due to high CV mortality, and shorter lifespan if compare with the Western society amid the absence of the established monitoring or screening system for major CV risk factors. The purpose of this systematic epidemiological update was to explore CV burden in Russia. The study represents pooled results with a systematic epidemiological review of the national mass screening, selected randomized clinical trials and statistical datasets of the national public health CV institutions exploring the trends of the CV burden in all 83 regions of Russia. We overviewed data from a number of the available Russian-speaking national data sources of 2001-2014, and NANOM-FIM trial (NCT01270139) as the only available real-world population study. The CV diseases in Russia accounted for 54.9% of all deaths in 2011-2014. The death rate was 13.3 per 1,000 citizens with CV mortality of 653.9 per 100,000. The life expectancy achieves 64.3 years for male and 76.1 years for female. The mean age of pts in trial was 51.6 years (77.2% males). A total of 175 Russian PCI centers implemented 205,902 angio a year, and 75,378 PCI achieving 531 PCI per 1,000,000 with placement of 101,451 stents (1.37 stents per PCI; 48,057 DES). The smoking (17.3% of screened with a 2,786 cigarettes a year; 70.6% in trial), excessive alcohol consumption (1.8% of screened with a 11.6 L per year; 50.6% in trial), unhealthy Russian diet (abundance of carbohydrates/sugar, saturated and trans fats in 24.3% of screened), psychosocial factors (20%) and physical inactivity (19.6% of screened) remain the major modifiable risk factors. They, in turn, affect such risk factors as dyslipidemia (86.7% in trial), obesity (16.7% of screened; BMI in trial was 28.4), and hypertension (40.8% suffered; 86.1% in trial). CV mortality was not directly associated with a level of poverty (r=0.26, P=0.02) or socio-economic development (P>0.05) in regions. The documented 27% 10-year decline of CV mortality was interpreted as a success of the national policy. Mortality statistics show the stark reality of a high CV burden in Russia. New national program and aggressive emerging efforts are required to tackle CV diseases in Russia.


A comparison of two methods of determining colour change in the assessment of the quality of pork.
The colour of pork is one of the most important measures of meat quality. For the consumer, it is a sign of freshness, crucial for the decision to buy a product. The storage of meat results in constant col- our changes. Differences can be found in all colour parameters, especially chromatic ones, such as redness (a*) and hue angle (h°), as these changes are significantly correlated with a number of quality traits. Colour change can be determined not only by evaluating changes in colour parameters, but also by establishing the change in absorbance of specific wavelengths of light. The aim of this study was to compare the usefulness of these two methods of determining colour change: parameters measured on the CIELAB and CIELCh scales and colour change (%CC) determined based on the absorbance of selected wavelengths of light, i.e. 505, 540, 560, 580 and 630 nm. In this paper, we analysed two methods of measuring colour change in terms of their relevance for meat quality assessment. The study involved 150 samples of the longissimus lumborum muscle collected from 150 carcasses of pigs slaughtered on an industrial line, weighing 65-107 kg, with an average carcass weight of 86.1 kg. Meat sensory analysis (colour, wateriness, firmness), physicochemical assessment (colour, WHC, pH48) and proximate analysis (moisture, total protein, fat, ash) were carried out. The methods included the determination of (I) changes in colour parameters measured on the CIELAB and CIELCh scales, and (II) colour change (%CC) based on the absorbance of selected wavelengths: 505, 540, 560, 580 and 630 nm. Chromatic absorbance values of a wavelength of 525 nm (A525p) and the relative content of Mb, MbO2 and MetMb were calculated according to the method proposed by Krzywicki (1979). Basic chemical composition - with few exceptions - was not significantly correlated with differenc- es in the values of colour parameters (ΔL*, ?a*, ?b*, ΔC*, Δh°) and colour change (%CC). With an increase in scores in the sensory evaluation of colour, wateriness, and firmness, and an increase in WHC and pH48, the differences in lightness (ΔL*) significantly increased and differences in chromatic value (?a*, ?b*, ΔC*, Δh°) and %CC decreased, for all tested combinations of wavelengths. The highest correlation coefficients were observed for ?a* and Δh° and %CC540/630 and %CC580/630, with colour change (%CC540/630 and %CC580/630) showing a stronger correlation with the tested meat quality traits than the differences in redness (?a*) and hue angle (Δh°). We found a very strong correlation between changes in the relative levels of MetMb (ΔMetMb) and colour change (%CC) based on the absorbance of wavelengths of 540 and 505 nm, and 580 and 505 nm (r = 0.950*** and r = 0.967***, respectively). The determination of colour change based on the absorbance of two combinations of wave- lengths, i.e. (I) 540 and 630, and (II) 580 nm and 630 nm, is more useful in the assessment of pork quality than changes in two most relevant parameters of the CIELAB and CIELCh scales, i.e. redness (Δa*) and hue angle (Δh°). Moreover, the determination of absorbance of a wavelength of 630 nm was more useful for determin- ing colour change (%CC) than 505 nm. Colour change (%CC) based on the absorbance of wavelengths of 540 and 505 nm and 580 and 505 nm reflect the changes in the relative amounts of MetMb very well.


Assessment and quality control of incontinence care in long-term nursing facilities.
A statistical quality-control process was used to assess how well incontinence management procedures were being implemented by indigenous nursing staff in four nursing homes. Eighty-one incontinent patients were treated with the prompted-voiding toileting procedure. Thirty-six of these patients proved responsive to the toileting procedures, and nursing home staff was instructed to maintain the toileting program for these responsive patients. The first part of the quality-control model involved setting job standards specifying how dry the patients should be if toileted on a 2-hour schedule. Second, a job-monitoring control chart was used to continuously assess how well the job standards were being met. The remaining forty-five patients, who were unresponsive to the toileting protocol, were managed with a 2-hour changing schedule. Job standards specifying how wet (volume) the patient would be if changed on a two-hour basis were set. Control-chart monitoring of these patients urine output assessed how well the changing procedures were being implemented. Incontinence care in nursing homes is difficult to supervise because of problems in measuring how consistently nursing aides change or toilet patients. This paper describes a management system for effective incontinence care.


Effects of accidental chlorine inhalation on pulmonary function.
In an industrial accident, 19 previously healthy workers were briefly exposed to high concentrations of chlorine gas. Pulmonary function tests were done at intervals for about two years but complete follow-up data were available in only 11 subjects. Immediately following the exposure, airway obstruction was detected in 10 of 19 patients; 700 days later this was found in only 3 of 11 patients. Two of these three patients had a history of smoking, however. The mean residual volume was 141% +/- 97 (mean +/- standard error of the mean) on day 1. In subsequent follow-up studies, the residual volume progressively fell in all patients, and 700 days later the mean residual volume was 90% +/- 5. In 5 of the 19 subjects, all pulmonary function test results were within normal limits on day 1. Apparently in some subjects acute exposure to chlorine gas may cause immediate changes in the lung functions, but these changes gradually resolve. Because of the small number of patients in our series, however, the long-term effects of chlorine are less apparent.


Three-dimensional transoesophageal echocardiography: how to use and when to use-a clinical consensus statement from the European Association of Cardiovascular Imaging of the European Society of Cardiology.
Three-dimensional transoesophageal echocardiography (3D TOE) has been rapidly developed in the last 15 years. Currently, 3D TOE is particularly useful as an additional imaging modality for the cardiac echocardiographers in the echo-lab, for cardiac interventionalists as a tool to guide complex catheter-based procedures cardiac, for surgeons to plan surgical strategies, and for cardiac anaesthesiologists and/or cardiologists, to assess intra-operative results. The authors of this document believe that acquiring 3D data set should become a 'standard part' of the TOE examination. This document provides (i) a basic understanding of the physic of 3D TOE technology which enables the echocardiographer to obtain new skills necessary to acquire, manipulate, and interpret 3D data sets, (ii) a description of valvular pathologies, and (iii) a description of non-valvular pathologies in which 3D TOE has shown to be a diagnostic tool particularly valuable. This document has a new format: instead of figures randomly positioned through the text, it has been organized in tables which include figures. We believe that this arrangement makes easier the lecture by clinical cardiologists and practising echocardiographers.


Analysis of Global Transcriptome Change in Mouse Embryonic Fibroblasts After dsDNA and dsRNA Viral Mimic Stimulation.
The activation of innate immunity by viral nucleic acids present in the cytoplasm plays an essential role in controlling viral infection in both immune and non-immune cells. The dsDNA and dsRNA viral mimics can stimulate the cytosolic nucleic acids sensors and activate the antiviral innate immunity. In this study, taking advantage of dsDNA and dsRNA viral mimics, we investigated the global transcriptome changes after the antiviral immunity activation in mouse embryonic fibroblasts. Results from our data identified a positive feedback up-regulation of sensors (e.g., Tlr2, Tlr3, Ddx58, cGAS), transducers (e.g., Traf2, Tbk1) and transcription factors (e.g., Irf7, Jun, Stat1, Stat2) in multiple pathways involved in detecting viral or microbial infections upon viral mimic stimulation. A group of genes involved in DNA damage response and DNA repair such as Parp9, Dtx3l, Rad52 were also up-regulated, implying the involvement of these genes in antiviral immunity. Molecular function analysis further showed that groups of helicase genes (e.g., Dhx58, Helz2), nuclease genes (e.g., Dnase1l3, Rsph10b), methyltransferase genes (e.g., histone methyltransferase Prdm9, Setdb2; RNA methyltransferase Mettl3, Mttl14), and protein ubiquitin-ligase genes (e.g., Trim genes and Rnf genes) were up-regulated upon antiviral immunity activation. In contrast, viral mimic stimulation down-regulated genes involved in a broad range of general biological processes (e.g., cell division, metabolism), cellular components (e.g., mitochondria and ribosome), and molecular functions (e.g., cell-cell adhesion, microtubule binding). In summary, our study provides valuable information about the global transcriptome changes upon antiviral immunity activation. The identification of novel groups of genes up-regulated upon antiviral immunity activation serves as useful resource for mining new antiviral sensors and effectors.


Angle closure in the Namil study in central South Korea.
To assess the prevalence and associated risk factors of angle closure in a defined population as part of the Namil Study. In this cross-sectional epidemiologic study for residents aged 40 years or older in Namil-myon, a rural area in central South Korea, the examination included slitlamp biomicroscopy, applanation tonometry, gonioscopy, autorefraction, fundus photography, corneal thickness measurement, visual field test with frequency-doubling technology, and anterior chamber depth (ACD) and axial length (AL) measurements with partial coherence interferometry. Standard automated field test and optical coherence tomography or scanning laser polarimetry were performed to confirm the glaucomatous visual field/optic disc damage. Angle closure included primary angle-closure suspect (PACS), primary angle closure (PAC), and primary angle-closure glaucoma (PACG). Definitions of PACS, PAC, and PACG were based on the recommendations from the International Society for Geographical &Epidemiological Ophthalmology. Among the 1426 individuals enrolled for the assessment, with exclusion of cataract surgery, the prevalence rates of PACS, PAC, PACG, and overall angle closure in at least 1 eye were 2.0% (95% CI, 1.3%-2.8%), 0.5% (95% CI, 0.1%-0.9%), 0.7% (95% CI, 0.3%-1.1%), and 3.2% (95% CI, 2.3%-4.2%), respectively. Multivariate analysis found that older age (odds ratio [OR], 1.8797; 95% CI, 1.4624-2.4162), shallower ACD (OR, 0.9982; 95% CI, 0.9977-0.9987), and shorter AL (OR 0.9978; 95% CI, 0.9969-0.9988) (P < .001 for each) were significantly associated with angle closure. The overall prevalence of angle closure was 3.2% in the present study. On the basis of these findings, increasing age, shallower ACD, and shorter AL appear to be associated with angle closure. TRIAL REGISTRATION clinicaltrials.gov Identifier: NCT00727168.


VEX1 controls the allelic exclusion required for antigenic variation in trypanosomes.
Allelic exclusion underpins antigenic variation and immune evasion in African trypanosomes. These bloodstream parasites use RNA polymerase-I (pol-I) to transcribe just one telomeric variant surface glycoprotein (VSG) gene at a time, producing superabundant and switchable VSG coats. We identified trypanosome VSG exclusion-1 (VEX1) using a genetic screen for defects in telomere-exclusive expression. VEX1 was sequestered by the active VSG and silencing of other VSGs failed when VEX1 was either ectopically expressed or depleted, indicating positive and negative regulation, respectively. Positive regulation affected VSGs and nontelomeric pol-I-transcribed genes, whereas negative regulation primarily affected VSGs. Negative regulation by VEX1 also affected telomeric pol-I-transcribed reporter constructs, but only when they contained blocks of sequence sharing homology with a pol-I-transcribed locus. We conclude that restricted positive regulation due to VEX1 sequestration, combined with VEX1-dependent, possibly homology-dependent silencing, drives a "winner-takes-all" mechanism of allelic exclusion.


Phosphopeptide enrichment by covalent chromatography after derivatization of protein digests immobilized on reversed-phase supports.
A rugged sample-preparation method for comprehensive affinity enrichment of phosphopeptides from protein digests has been developed. The method uses a series of chemical reactions to incorporate efficiently and specifically a thiol-functionalized affinity tag into the analyte by barium hydroxide catalyzed β-elimination with Michael addition using 2-aminoethanethiol as nucleophile and subsequent thiolation of the resulting amino group with sulfosuccinimidyl-2-(biotinamido) ethyl-1,3-dithiopropionate. Gentle oxidation of cysteine residues, followed by acetylation of α- and ε-amino groups before these reactions, ensured selectivity of reversible capture of the modified phosphopeptides by covalent chromatography on activated thiol sepharose. The use of C18 reversed-phase supports as a miniaturized reaction bed facilitated optimization of the individual modification steps for throughput and completeness of derivatization. Reagents were exchanged directly on the supports, eliminating sample transfer between the reaction steps and thus, allowing the immobilized analyte to be carried through the multistep reaction scheme with minimal sample loss. The use of this sample-preparation method for phosphopeptide enrichment was demonstrated with low-level amounts of in-gel-digested protein. As applied to tryptic digests of α-S1- and β-casein, the method enabled the enrichment and detection of the phosphorylated peptides contained in the mixture, including the tetraphosphorylated species of β-casein, which has escaped chemical procedures reported previously. The isolates proved highly suitable for mapping the sites of phosphorylation by collisionally induced dissociation. β-Elimination, with consecutive Michael addition, expanded the use of the solid-phase-based enrichment strategy to phosphothreonyl peptides and to phosphoseryl/phosphothreonyl peptides derived from proline-directed kinase substrates and to their O-sulfono- and O-linked β-N-acetylglucosamine (O-GlcNAc)-modified counterparts. Solid-phase enzymatic dephosphorylation proved to be a viable tool to condition O-GlcNAcylated peptide in mixtures with phosphopeptides for selective affinity purification. Acetylation, as an integral step of the sample-preparation method, precluded reduction in recovery of the thiolation substrate caused by intrapeptide lysine-dehydroalanine cross-link formation. The solid-phase analytical platform provides robustness and simplicity of operation using equipment readily available in most biological laboratories and is expected to accommodate additional chemistries to expand the scope of solid-phase serial derivatization for protein structural characterization.


Tileable BTF.
This paper presents a modular framework to efficiently apply the bidirectional texture functions (BTF) onto object surfaces. The basic building blocks are the BTF tiles. By constructing one set of BTF tiles, a wide variety of objects can be textured seamlessly without re-synthesizing the BTF. The proposed framework nicely decouples the surface appearance from the geometry. With this appearance-geometry decoupling, one can build a library of BTF tile sets to instantaneously dress and render various objects under variable lighting and viewing conditions. The core of our framework is a novel method for synthesizing seamless high-dimensional BTF tiles, that are difficult for existing synthesis techniques. Its key is to shorten the cutting paths and broaden the choices of samples so as to increase the chance of synthesizing seamless BTF tiles. To tackle the enormous data, the tile synthesis process is performed in compressed domain. This not just allows the handling of large BTF data during the synthesis, but also facilitates compact storage of the BTF in GPU memory during the rendering.


Oligodendrocyte progenitor cell numbers and migration are regulated by the zebrafish orthologs of the NF1 tumor suppressor gene.
Neurofibromatosis type 1 is the most commonly inherited human cancer predisposition syndrome. Neurofibromin (NF1) gene mutations lead to increased risk of neurofibromas, schwannomas, low grade, pilocytic optic pathway gliomas, as well as malignant peripheral nerve sheath tumors and glioblastomas. Despite the evidence for NF1 tumor suppressor function in glial cell tumors, the mechanisms underlying transformation remain poorly understood. In this report, we used morpholinos to knockdown the two nf1 orthologs in zebrafish and show that oligodendrocyte progenitor cell (OPC) numbers are increased in the developing spinal cord, whereas neurons are unaffected. The increased OPC numbers in nf1 morphants resulted from increased proliferation, as detected by increased BrdU labeling, whereas TUNEL staining for apoptotic cells was unaffected. This phenotype could be rescued by the forced expression of the GTPase-activating protein (GAP)-related domain of human NF1. In addition, the in vivo analysis of OPC migration following nf1 loss using time-lapse microscopy demonstrated that olig2-EGFP(+) OPCs exhibit enhanced cell migration within the developing spinal cord. OPCs pause intermittently as they migrate, and in nf1 knockdown animals, they covered greater distances due to a decrease in average pause duration, rather than an increase in velocity while in motion. Interestingly, nf1 knockdown also leads to an increase in ERK signaling, principally in the neurons of the spinal cord. Together, these results show that negative regulation of the Ras pathway through the GAP activity of NF1 limits OPC proliferation and motility during development, providing insight into the oncogenic mechanisms through which NF1 loss contributes to human glial tumors.


Osteopontin-enhanced hepatic metastasis of colorectal cancer cells.
Liver metastasis is a major cause of mortality from colorectal cancer (CRC). However, mechanisms underlying this process are largely unknown. Osteopontin (OPN) is a secreted phosphorylated glycoprotein that is involved in tumor migration and metastasis. The role of OPN in cancer is currently unclear. In this study, OPN mRNA was examined in tissues from CRC, adjacent normal mucosa, and liver metastatic lesions using quantitative real-time PCR analysis. The protein expression of OPN and its receptors (integrin αv and CD44 v6) was detected by using an immunohistochemical (IHC) method. The role of OPN in liver metastasis was studied in established colon cancer Colo-205 and SW-480 cell lines transfected with sense- or antisense-OPN eukaryotic expression plasmids by flow cytometry and cell adhesion assay. Fluorescence redistribution after photobleaching (FRAP) was used to study gap functional intercellular communication (GJIC) among OPN-transfected cells. It was found that OPN was highly expressed in metastatic hepatic lesions from CRC compared to primary CRC tissue and adjacent normal mucosa. The expression of OPN mRNA in tumor tissues was significantly related with the CRC stages. OPN expression was also detected in normal hepatocytes surrounding CRC metastatic lesions. Two known receptors of OPN, integrin αv and CD44v6 proteins, were strongly expressed in hepatocytes from normal liver. CRC cells with forced OPN expression exhibited increased heterotypic adhesion with endothelial cells and weakened intercellular communication. OPN plays a significant role in CRC metastasis to liver through interaction with its receptors in hepatocytes, decreased homotypic adhesion, and enhanced heterotypic adhesion.


The serotonin 1A receptor gene confer susceptibility to mood disorders: results from an extended meta-analysis of patients with major depression and bipolar disorder.
The serotonin 1A receptor gene (HTR1A) has been associated with mood disorders (MDs), including major depressive disorder (MDD) and bipolar disorder (BP). Therefore, we conducted a systematic review and meta-analysis between rs6295 (C-1019G) as well as rs878567 in HTR1A and MDs. Searching PubMed through May 2012, 15 studies, including our own, previously unpublished association study (135 MDD patients and 107 healthy controls), met inclusion criteria for the meta-analysis of rs6295 (4,297 MDs patients and 5,435 controls). Five association studies met criteria for the meta-analysis of rs878567 (2041MDs patients and 2,734 controls). rs6295 was associated with combined MDs (P allele model = 0.007 and P recessive model = 0.01). When divided by diagnostic subgroup (MDD = 3,119 patients and 4,380 controls or BP = 1,170 patients and 2,252 controls), rs6295 was associated with each MDs separately (MDD: P allele model = 0.006, P recessive model = 0.01; BP: P dominant model = 0.003). Likewise, rs878567 was associated with combined MDs (2,041 patients and 2,734 controls (P allele model = 0.0002, P dominant model = 0.0008, and P recessive model = 0.01). When divided by diagnostic subgroup (MDD = 1,013 patients and 1,728 controls or BP = 1,051 patients and 2,099 controls), rs878567 was associated with MDD (P allele model = 0.0007 and P dominant model = 0.01), while only one BP study had such data, precluding a meta-analysis. All of these significances survived correction for multiple comparisons. Results from this expanded meta-analysis, which included our own new study, suggest that rs6295 (C-1019G) and rs878567 in HTR1A are related to the pathophysiology of MDs, with overlap between MDD and BP. Findings provide additional clues to the underlying biology and treatment targets in MDs.


A nonstructural viral protein expressed by a recombinant vaccinia virus protects against lethal cytomegalovirus infection.
The nonstructural immediate-early protein pp89 of murine cytomegalovirus (MCMV) is the first viral protein synthesized after infection and has a regulatory function in viral gene expression. Despite its localization in the nucleus of infected cells, pp89 is also the dominant antigen recognized by MCMV-specific cytolytic T lymphocytes. The recombinant vaccinia virus MCMV-ieI-VAC, which expresses pp89, was used to study the capacity of this protein to induce protective immunity in BALB/c mice. Vaccination with MCMV-ieI-VAC induced a long-lasting immunity that protected mice against challenge with a lethal dose of MCMV but did not prevent infection and morbidity. In vivo depletion of CD8+ T lymphocytes before challenge completely abrogated the protective immunity. CD8+ T lymphocytes derived from MCMV-ieI-VAC-primed donors and adoptively transferred into sublethally irradiated and MCMV-infected recipients were found to limit viral replication in host tissues, whereas CD4+ T lymphocytes and pp89-specific antiserum had no protective effect. The data demonstrate for the first time that a single nonstructural viral protein can confer protection against a lethal cytolytic infection and that this immunity is entirely mediated by the CD8+ subpopulation of T lymphocytes.


Affect dynamics, affective forecasting, and aging.
Affective forecasting, experienced affect, and recalled affect were compared in younger and older adults during a task in which participants worked to win and avoid losing small monetary sums. Dynamic changes in affect were measured along valence and arousal dimensions, with probes during both anticipatory and consummatory task phases. Older and younger adults displayed distinct patterns of affect dynamics. Younger adults reported increased negative arousal during loss anticipation and positive arousal during gain anticipation. In contrast, older adults reported increased positive arousal during gain anticipation but showed no increase in negative arousal on trials involving loss anticipation. Additionally, younger adults reported large increases in valence after avoiding an anticipated loss, but older adults did not. Younger, but not older, adults exhibited forecasting errors on the arousal dimension, underestimating increases in arousal during anticipation of gains and losses and overestimating increases in arousal in response to gain outcomes. Overall, the findings are consistent with a growing literature suggesting that older people experience less negative emotion than their younger counterparts and further suggest that they may better predict dynamic changes in affect.


Respiratory illness in children: what makes parents decide to consult?
Parents of a stratified random sample of 234 children from 21 training practices in north east England were interviewed at home. All these children had been reported in a postal questionnaire as having had a cough between six and 10 weeks before the interview. Interviews covered social characteristics of the family, the severity of the child's cough and the child's previous respiratory history. These data were analysed using the statistical technique of logistic regression. This produced a good model of consulting behaviour. The model showed that a doctor was likely to be consulted if the child had severe symptoms, or if the cough affected the child's behaviour. This suggests that most parents deciding whether to consult the doctor make careful decisions based on what they see as objective criteria. No social characteristic had a significant influence on the decision to consult the doctor over and above the influence of the characteristics and effect of the cough itself.


Links between methanotroph community composition and CH oxidation in a pine forest soil.
The main gap in our knowledge about what determines the rate of CH(4) oxidation in forest soils is the biology of the microorganisms involved, the identity of which remains unclear. In this study, we used stable-isotope probing (SIP) following (13)CH(4) incorporation into phospholipid fatty acids (PLFAs) and DNA/RNA, and sequencing of methane mono-oxygenase (pmoA) genes, to identify the influence of variation in community composition on CH(4) oxidation rates. The rates of (13)C incorporation into PLFAs differed between horizons, with low (13)C incorporation in the organic soil and relatively high (13)C incorporation into the two mineral horizons. The microbial community composition of the methanotrophs incorporating the (13)C label also differed between horizons, and statistical analyses suggested that the methanotroph community composition was a major cause of variation in CH(4) oxidation rates. Both PLFA and pmoA-based data indicated that CH(4) oxidizers in this soil belong to the uncultivated 'upland soil cluster alpha'. CH(4) oxidation potential exhibited the opposite pattern to (13)C incorporation, suggesting that CH(4) oxidation potential assays may correlate poorly with in situ oxidation rates. The DNA/RNA-SIP assay was not successful, most likely due to insufficient (13)C-incorporation into DNA/RNA. The limitations of the technique are briefly discussed.


Clinical effectiveness of a rehabilitation program integrating exercise, self-management, and active coping strategies for chronic knee pain: a cluster randomized trial.
Chronic knee pain is a major cause of disability and health care expenditure, but there are concerns about efficacy, cost, and side effects associated with usual primary care. Conservative rehabilitation may offer a safe, effective, affordable alternative. We compared the effectiveness of a rehabilitation program integrating exercise, self-management, and active coping strategies (Enabling Self-management and Coping with Arthritic Knee Pain through Exercise [ESCAPE-knee pain]) with usual primary care in improving functioning in persons with chronic knee pain. We conducted a single-blind, pragmatic, cluster randomized controlled trial. Participants age >/=50 years, reporting knee pain for >6 months, were recruited from 54 inner-city primary care practices. Primary care practices were randomized to continued usual primary care (i.e., whatever intervention a participant's primary care physician deemed appropriate), usual primary care plus the rehabilitation program delivered to individual participants, or usual primary care plus the rehabilitation program delivered to groups of 8 participants. The primary outcome was self-reported functioning (Western Ontario and McMaster Universities Osteoarthritis Index physical functioning [WOMAC-func]) 6 months after completing rehabilitation. A total of 418 participants were recruited; 76 (18%) withdrew, only 5 (1%) due to adverse events. Rehabilitated participants had better functioning than participants continuing usual primary care (-3.33 difference in WOMAC-func score; 95% confidence interval [95% CI] -5.88, -0.78; P = 0.01). Improvements were similar whether participants received individual rehabilitation (-3.53; 95% CI -6.52, -0.55) or group rehabilitation (-3.16; 95% CI -6.55, -0.12). ESCAPE-knee pain provides a safe, relatively brief intervention for chronic knee pain that is equally effective whether delivered to individuals or groups of participants.


Endothelin subtypes: effect on isolated rhesus monkey ciliary muscle.
The effects of endothelin (ET)-1, ET-2, ET-3 and sarafotoxin S6C on the contractile response in the longitudinal and coronal vectors of the isolated rhesus monkey ciliary muscle were studied. Fresh ciliary muscle strips from young and middleaged rhesus monkeys were mounted in an apparatus capable of monitoring contractile force simultaneously in the two vectors. The responses to the ET compounds were measured and compared to those produced with 1 microM carbachol. ET-1 produced contractions of up to 15% of the near-maximum response to carbachol in both vectors of 66% of ciliary muscle strips studied. In responsive strips, the maximal ET-1 induced contraction was approximately equal in both vectors, but the longitudinal vector was approximately 5-fold more sensitive than the circular. All ciliary muscle strips responded reproducibly to carbachol. None of the other ET compounds tested had any effect, suggesting that the ETA receptor may predominate in rhesus monkey ciliary muscle. Because ET-1 induces only weak and interindividually variable contraction in isolated rhesus monkey ciliary muscle strips, ET-1's enhancement of outflow facility with only minimal induction of accommodation in the living monkey may be due to effects directly on the trabecular meshwork. However, the 5-fold greater potency of ET-1 in the longitudinal compared to the circular contractile vector may indicate that selective contraction of the longitudinal portion of the ciliary muscle with consequent deformation of the trabecular meshwork but not the lens is also involved.


Handwriting movement analyses for monitoring drug-induced motor side effects in schizophrenia patients treated with risperidone.
Epidemiologic studies indicate that nearly 60% of schizophrenia (SZ) patients treated with conventional antipsychotic drugs develop extrapyramidal side effects (EPS) such as parkinsonism and tardive dyskinesia. Although the prevalence of EPS has decreased due to the newer antipsychotics, EPS continue to limit the effectiveness of these medicines. Ongoing monitoring of EPS is likely to improve treatment outcome or compliance and reduce the frequency of re-hospitalization. A quantitative analysis of handwriting kinematics was used to evaluate effects of antipsychotic medication type and dose in schizophrenia patients. Twenty-seven schizophrenia patients treated with risperidone, six schizophrenia patients who received no antipsychotic medication and 47 healthy comparison participants were enrolled. Participants performed a 20-min handwriting task consisting of loops of various sizes and a sentence. Data were captured and analyzed using MovAlyzeR software. Results indicated that risperidone-treated participants exhibited significantly more dysfluent handwriting movements than either healthy or untreated SZ participants. Risperidone-treated participants exhibited lower movement velocities during production of simple loops compared to unmedicated patients. Handwriting dysfluency during sentence writing increased with dose. A 3-factor model consisting of kinematic variables derived from sentence writing accounted for 83% (r=.91) of the variability in medication dose. In contrast, we found no association between observer-based EPS severity ratings and medication dose. These findings support the importance of handwriting-based measures to monitor EPS in medicated schizophrenia patients.


Differential regulation of two sets of mesonephric tubules by WT-1.
Mammalian renal development undergoes two transient stages, the pronephros and the mesonephros. While the regulation of metanephric differentiation has received considerable attention, very little is known about the mode of differentiation of the mesonephros and its regulation. We have followed mesonephric differentiation to unravel the developmental mechanisms and fates of mesonephric tubules by whole-mount immunohistology using antibodies to laminin, brush border epitopes, cytokeratin-8/18, p75 neurotrophin receptor and some other renal antigens as markers. In rat and mouse embryos, two distinct sets of tubules were observed throughout mesonephric development. Four to six pairs of cranial mesonephric tubules developed as outgrowths from the Wolffian duct. The majority of tubules were caudal tubules which never fused with the Wolffian and differentiated similarly to metanephric nephrons. The murine mesonephric tubules degenerate by apoptosis, except in males where the cranial tubules become the epididymal ducts. These developmental differences between the cranial and caudal sets of tubules suggested different regulatory systems for each. Targeted disruption of the Wilms' tumour gene product, WT-1, results in renal aplasia, and a reduction in the number of mesonephric tubules (Kreidberg, J. A., Sariola, H., Loring, J., Maeda, M., Pelletier, J., Housman, D. and Jaenisch, R. (1993). Cell 74, 679-691). We therefore analysed more closely mesonephric differentiation in WT-1-deficient mice, and showed that they only develop the cranial mesonephric tubules but not the caudal ones. Thus, WT-1 appears to regulate only the development of the caudal mesonephric tubules that conceivably are formed from mesenchymal cells like the metanephric tubules. WT-1 therefore seems to be necessary for the mesenchyme to epithelium transitions at different stages of nephrogenesis.


Trainer-to-student ratios for teaching psychomotor skills in health care fields, as applied to osteopathic manipulative medicine.
The hallmark of osteopathic medical education is the inclusion of hands-on instruction in osteopathic manipulative medicine (OMM), which includes palpatory diagnosis and osteopathic manipulative treatment (OMT). This OMM training typically involves a primary instructor presenting theory and techniques with step-by-step demonstrations to a large group of first- and second-year osteopathic medical students. Additional instructors, referred to as table trainers, assist the primary instructor by supervising the students as they practice the presented techniques. To the authors' knowledge, there is no currently accepted standard for a table trainer-to-student ratio in OMM skills laboratories within osteopathic medical schools in the United States. However, through a Google Web search and PubMed literature review, the authors identified published trainer-to-student ratios used in other health care skills training curricula. Psychomotor skills training courses in health care fields typically have a table trainer-to-student ratio of 1 trainer to 8 or fewer students. On the basis of these findings and psychomotor skills learning theory, the authors conclude that this ratio is likely sufficient for OMM skills training.


O vacancy formation in (Pr/Gd)BaCo2O5.5 and the role of antisite defects.
In search for materials for intermediate temperature solid oxide fuel cells, (Pr/Gd)BaCo2O5.5 is investigated by first principles calculations. Antisite defects are considered as they may modify the electronic and O diffusion properties but are rarely studied in double perovskite oxides. Octahedrally coordinated Co atoms are shown to realize intermediate and high spin states for PrBaCo2O5.5 and GdBaCo2O5.5, respectively, while pyramidally coordinated Co atoms always have high spin. It turns out that O vacancy formation is significantly easier in PrBaCo2O5.5 than in GdBaCo2O5.5, the difference in formation energy being hardly modified by antisite defects. While pyramidally coordinated Co atoms are not affected, we show that the presence of antisite defects causes parts of the octahedrally coordinated Co atoms to switch from intermediate to high spin.


Dynamics of the regulation of Hsp90 by the co-chaperone Sti1.
In eukaryotic cells, Hsp90 chaperones assist late folding steps of many regulatory protein clients by a complex ATPase cycle. Binding of clients to Hsp90 requires prior interaction with Hsp70 and a transfer reaction that is mediated by the co-chaperone Sti1/Hop. Sti1 furthers client transfer by inhibiting Hsp90's ATPase activity. To better understand how Sti1 prepares Hsp90 for client acceptance, we characterized the interacting domains and analysed how Hsp90 and Sti1 mutually influence their conformational dynamics using hydrogen exchange mass spectrometry. Sti1 stabilizes several regions in all three domains of Hsp90 and slows down dissociation of the Hsp90 dimer. Our data suggest that Sti1 inhibits Hsp90's ATPase activity by preventing N-terminal dimerization and docking of the N-terminal domain with the middle domain. Using crosslinking and mass spectrometry we identified Sti1 segments, which are in close vicinity of the N-terminal domain of Hsp90. We found that the length of the linker between C-terminal dimerization domain and the C-terminal MEEVD motif is important for Sti1 association rates and propose a kinetic model for Sti1 binding to Hsp90.


Kinematics of the trapeziometacarpal joint after sectioning of ligaments.
This study was designed to examine the roles of ligaments in the maintenance of the articular kinematics of the trapeziometacarpal joint. Circumduction of the trapeziometacarpal joint was studied in 12 hands from fresh human cadavera. With use of a magnetic tracking system, changes in the motion of the base of the first metacarpal after ligament sectioning were analyzed and compared with those of the normal joint. Two sets of ligaments were sectioned: (a) the anterior oblique and ulnar collateral ligaments and (b) the first intermetacarpal ligament and the ulnar joint capsule. Sectioning of the anterior oblique and ulnar collateral ligaments resulted in a significant dorsal-ulnar shift in the path of the base of the first metacarpal. However, sectioning of the first intermetacarpal ligament did not affect the movement pattern of the center of the base. The anterior oblique and ulnar collateral ligaments provided constraint of the trapeziometacarpal joint during circumduction of the thumb.


A Meshfree Representation for Cardiac Medical Image Computing.
The prominent advantage of meshfree method, is the way to build the representation of computational domain, based on the nodal points without any explicit meshing connectivity. Therefore, meshfree method can conveniently process the numerical computation inside interested domains with large deformation or inhomogeneity. In this paper, we adopt the idea of meshfree representation into cardiac medical image analysis in order to overcome the difficulties caused by large deformation and inhomogeneous materials of the heart. In our implementation, as element-free Galerkin method can efficiently build a meshfree representation using its shape function with moving least square fitting, we apply this meshfree method to handle large deformation or inhomogeneity for solving cardiac segmentation and motion tracking problems. We evaluate the performance of meshfree representation on a synthetic heart data and an in-vivo cardiac MRI image sequence. Results showed that the error of our framework against the ground truth was 0.1189 ± 0.0672 while the error of the traditional FEM was 0.1793 ± 0.1166. The proposed framework has minimal consistency constraints, handling large deformation and material discontinuities are simple and efficient, and it provides a way to avoid the complicated meshing procedures while preserving the accuracy with a relatively small number of nodes.


Enhancer-dependent transcriptional oscillations in mouse erythroleukemia cells.
By using recombinase-mediated cassette exchange, a method that allows integration of single copies of different constructs at the same predetermined chromosomal location, several expression cassettes have been integrated at a randomly chosen locus in the genome of mouse erythroleukemia cells. The cassettes studied contain the human beta-globin promoter fused to lacZ coding sequences either alone or linked to DNase I-hypersensitive site HS2, HS3, or HS234 (a large locus control region fragment containing HS2, HS3, and HS4) of the human beta-globin locus control region. Analysis of expression of these cassettes revealed mosaic expression patterns reminiscent of, but clearly different from, position effect variegation. Further investigations demonstrated that these mosaic expression patterns are caused by dynamic activation and inactivation of the transcription unit, resulting in oscillations of expression. These oscillations occur once in every few cell cycles at a rate specific for the enhancer present at the locus. DNase I sensitivity studies revealed that the chromatin is accessible and that DNase-hypersensitive sites were present whether or not the transcription unit is active, suggesting that the oscillations occur between transcriptionally competent and transcriptionally active chromatin conformations, rather than between open and closed chromatin conformations. Treatment of oscillating cells with trichostatin A eliminates the oscillations only after the cells have passed through late G1 or early S, suggesting that these oscillations might be caused by changes in histone acetylation patterns.


Effect of Melatonin on the Outcome of Assisted Reproductive Technique Cycles in Women with Diminished Ovarian Reserve: A Double-Blinded Randomized Clinical Trial.
Diminished ovarian reserve (DOR) significantly decreases the success rate of the assisted reproductive technique (ART). In this study, we assessed the effect of melatonin on the ART outcomes in women with DOR. A double-blinded, randomized, clinical trial was performed on 80 women with DOR as a pilot study in Shiraz, between 2014 and 2015. DOR was defined as the presence of 2 of the following 3 criteria: 1) anti-Müllerian hormone ≤1, 2) follicle-stimulating hormone ≥10, and 3) bilateral antral follicle count ≤6. The women received 3 mg/d melatonin or a placebo since the fifth day of one cycle prior to gonadotropin stimulation and continued the treatment up to the time of ovum pickup. The ART outcomes were compared between the groups using SPSS software. Finally, there were 32 women in the case and 34 in the placebo groups. The mean age and basal ovarian reserve test were the same between the groups. The serum estradiol level on the triggering day was significantly higher in the case group (P=0.005). The mean number of MII oocytes was higher in the case group, but the difference did not reach statistical significance. Number of the patients who had mature MII oocytes (P=0.014), top-quality embryos with grade 1 (P=0.049), and embryos with grades 1 and 2 (P=0.014) was higher among the women who received melatonin. However, the other ART outcomes were not different between the groups. The serum estradiol level was higher and more women with DOR had good-quality oocytes and embryos after receiving melatonin; however, no other outcome was different between the case and control groups. Trial Registration Number: IRCT2014041417264N1.


COVID-19 and Italy: what next?
The spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has already taken on pandemic proportions, affecting over 100 countries in a matter of weeks. A global response to prepare health systems worldwide is imperative. Although containment measures in China have reduced new cases by more than 90%, this reduction is not the case elsewhere, and Italy has been particularly affected. There is now grave concern regarding the Italian national health system's capacity to effectively respond to the needs of patients who are infected and require intensive care for SARS-CoV-2 pneumonia. The percentage of patients in intensive care reported daily in Italy between March 1 and March 11, 2020, has consistently been between 9% and 11% of patients who are actively infected. The number of patients infected since Feb 21 in Italy closely follows an exponential trend. If this trend continues for 1 more week, there will be 30 000 infected patients. Intensive care units will then be at maximum capacity; up to 4000 hospital beds will be needed by mid-April, 2020. Our analysis might help political leaders and health authorities to allocate enough resources, including personnel, beds, and intensive care facilities, to manage the situation in the next few days and weeks. If the Italian outbreak follows a similar trend as in Hubei province, China, the number of newly infected patients could start to decrease within 3-4 days, departing from the exponential trend. However, this cannot currently be predicted because of differences between social distancing measures and the capacity to quickly build dedicated facilities in China.


Structural basis of chronic beryllium disease: linking allergic hypersensitivity and autoimmunity.
T-cell-mediated hypersensitivity to metal cations is common in humans. How the T cell antigen receptor (TCR) recognizes these cations bound to a major histocompatibility complex (MHC) protein and self-peptide is unknown. Individuals carrying the MHCII allele, HLA-DP2, are at risk for chronic beryllium disease (CBD), a debilitating inflammatory lung condition caused by the reaction of CD4 T cells to inhaled beryllium. Here, we show that the T cell ligand is created when a Be(2+) cation becomes buried in an HLA-DP2/peptide complex, where it is coordinated by both MHC and peptide acidic amino acids. Surprisingly, the TCR does not interact with the Be(2+) itself, but rather with surface changes induced by the firmly bound Be(2+) and an accompanying Na(+) cation. Thus, CBD, by creating a new antigen by indirectly modifying the structure of preexisting self MHC-peptide complex, lies on the border between allergic hypersensitivity and autoimmunity.


Chromosome-encoded beta-lactamases TUS-1 and MUS-1 from Myroides odoratus and Myroides odoratimimus (formerly Flavobacterium odoratum), new members of the lineage of molecular subclass B1 metalloenzymes.
Myroides odoratus and Myroides odoratimimus (formerly designated in a single species as Flavobacterium odoratum) are gram-negative aerobes and sources of nosocomial infections in humans. They have variable susceptibility to beta-lactams and a decreased susceptibility to carbapenems. Using genomic DNAs of M. odoratus CIP 103105 and M. odoratimimus CIP 103073 reference strains, shotgun cloning of beta-lactamase genes was performed, followed by protein expression in Escherichia coli. The deduced amino acid sequences of these beta-lactamase genes revealed that TUS-1 and MUS-1 from M. odoratus CIP 103105 and M. odoratimimus CIP 103073, respectively, shared 73% amino acid identity. Mature proteins TUS-1 and MUS-1, with pI values of 7.8 and 5.2, respectively, had relative molecular masses of ca. 26 kDa. These beta-lactamases are members of the subclass B1 of metallo-beta-lactamases and are distantly related to other metalloenzymes, being most closely related to IND-1 from Chryseobacterium indologenes (42% amino acid identity). However, phylogenic analysis showed that TUS-1 and MUS-1 belong to the same phylogenic lineage of subclass B1 enzymes that groups the subclass B1 beta-lactamases of Flavobacterium species. Kinetic parameters of purified beta-lactamases TUS-1 and MUS-1 detailed their hydrolysis spectra, which encompass most beta-lactams except aztreonam. beta-Lactamases TUS-1 and MUS-1 were classified in functional subgroup 3a of metalloenzymes. This work further characterizes chromosome-encoded metalloenzymes from Flavobacteriaceae species that explain at least part of their intrinsic resistance to beta-lactams.


Parallel Processing Method for Airborne Laser Scanning Data Using a PC Cluster and a Virtual Grid.
In this study, a parallel processing method using a PC cluster and a virtual grid is proposed for the fast processing of enormous amounts of airborne laser scanning (ALS) data. The method creates a raster digital surface model (DSM) by interpolating point data with inverse distance weighting (IDW), and produces a digital terrain model (DTM) by local minimum filtering of the DSM. To make a consistent comparison of performance between sequential and parallel processing approaches, the means of dealing with boundary data and of selecting interpolation centers were controlled for each processing node in parallel approach. To test the speedup, efficiency and linearity of the proposed algorithm, actual ALS data up to 134 million points were processed with a PC cluster consisting of one master node and eight slave nodes. The results showed that parallel processing provides better performance when the computational overhead, the number of processors, and the data size become large. It was verified that the proposed algorithm is a linear time operation and that the products obtained by parallel processing are identical to those produced by sequential processing.


The precipitation, growth and stability of mercury sulfide nanoparticles formed in the presence of marine dissolved organic matter.
The methylation of mercury is known to depend on the chemical forms of mercury (Hg) present in the environment and the methylating bacterial activity. In sulfidic sediments, under conditions of supersaturation with respect to metacinnabar, recent research has shown that mercury precipitates as β-HgS(s) nanoparticles (β-HgS(s)nano). Few studies have examined the precipitation of β-HgS(s)nano in the presence of marine dissolved organic matter (DOM). In this work, we used dynamic light scattering (DLS) coupled with UV-Vis spectroscopy and transmission electron microscopy (TEM) to investigate the formation and fate of β-HgS(s)nano formed in association with marine DOM extracted from the east and west of Long Island Sound, and at the shelf break of the North Atlantic Ocean, as well as with low molecular weight thiols. We found that while the β-HgS(s)nano formed in the presence of oceanic DOM doubled in size after 5 weeks, those forming in solutions with coastal DOM did not grow over time. In addition, when the HgII : DOM ratio was varied, β-HgS(s)nano only rapidly aggregated at high ratios (>41 μmol HgII per mg C) where the concentration of thiol groups was determined to be substantially low relative to HgII. This suggests that functional groups other than thiols could be involved in the stabilization of β-HgS(s)nano. Furthermore, we showed that β-HgS(s)nano forming under anoxic conditions remained stable and could therefore persist in the environment sufficiently to impact the methylation potential. Exposure of β-HgS(s)nano to sunlit and oxic environments, however, caused rapid aggregation and sedimentation of the nanoparticles, suggesting that photo-induced changes or oxidation of organic matter adsorbed on the surface of β-HgS(s)nano affected their stability in surface waters.


Insight in glioma susceptibility through an analysis of 6p22.3, 12p13.33-12.1, 17q22-23.2 and 18q23 SNP genotypes in familial and non-familial glioma.
The risk of glioma has consistently been shown to be increased twofold in relatives of patients with primary brain tumors (PBT). A recent genome-wide linkage study of glioma families provided evidence for a disease locus on 17q12-21.32, with the possibility of four additional risk loci at 6p22.3, 12p13.33-12.1, 17q22-23.2, and 18q23. To identify the underlying genetic variants responsible for the linkage signals, we compared the genotype frequencies of 5,122 SNPs mapping to these five regions in 88 glioma cases with and 1,100 cases without a family history of PBT (discovery study). An additional series of 84 familial and 903 non-familial cases were used to replicate associations. In the discovery study, 12 SNPs showed significant associations with family history of PBT (P < 0.001). In the replication study, two of the 12 SNPs were confirmed: 12p13.33-12.1 PRMT8 rs17780102 (P = 0.031) and 17q12-21.32 SPOP rs650461 (P = 0.025). In the combined analysis of discovery and replication studies, the strongest associations were attained at four SNPs: 12p13.33-12.1 PRMT8 rs17780102 (P = 0.0001), SOX5 rs7305773 (P = 0.0001) and STKY1 rs2418087 (P = 0.0003), and 17q12-21.32 SPOP rs6504618 (P = 0.0006). Further, a significant gene-dosage effect was found for increased risk of family history of PBT with these four SNPs in the combined data set (P(trend) <1.0 × 10(-8)). The results support the linkage finding that some loci in the 12p13.33-12.1 and 17q12-q21.32 may contribute to gliomagenesis and suggest potential target genes underscoring linkage signals.


Evaluation of drug effects on Toxoplasma gondii nuclear and plastid DNA replication using real-time PCR.
Toxoplasma gondii Nicolle and Manceaux, 1908 is a unicellular protozoan that can infect a broad spectrum of organisms including humans. In addition to a nuclear genome, it also carries a circular DNA within a plastid-like organelle (apicoplast) and a linear genome within its mitochondria. The plastid organelle has been shown to be the target of various anti-parasitic drugs or antibiotics. To evaluate the effects of agents on the DNA replication of T. gondii, we tested six drugs (ciprofloxacin, acetylspiramycin, clindamycin, azithromycin, artemether, and sulfadiazine) on the parasite cultured in Hela cells. After drug treatment for 48 h, the parasite growth and DNA replication were evaluated and quantitated using TaqMan real-time quantitative PCR with oligonucleotide primers synthesized based on a gene from the apicoplast genome (ycf24, Genbank accession no. U87145) and a gene from the nuclear genome (uprt, Genbank accession no. U10246). Our results showed that ciprofloxacin was the most effective in inhibiting the replication of the plastid DNA after 48 h drug treatment, with a reduction of 22% in the copy number of the plastid DNA. Artemether was the most effective drug in suppressing the proliferation of tachyzoites. This study also demonstrates that real-time quantitative PCR is a simple and useful technique for monitoring parasite growth and DNA replication.


Characterization of herpes simplex virus type-1 infection and herpetic stromal keratitis development in IFN-gamma knockout mice.
Herpetic stromal keratitis (HSK) has an immune-mediated pathogenesis that involves T cells that have a type 1 cytokine profile. IFN-gamma is suspected to be the type 1 cytokine involved in ocular pathology, and to test this notion more directly the pathogenesis of HSK was compared in mice deficient in the IFN-gamma gene (gamma knockout or gko) and control mice (wild-type littermates or BALB/c mice). The clinical course of HSK in gko mice closely paralleled that in control mice, yet virus persisted in the corneas of gko mice for an extended period of time, severe periocular skin lesions developed, and gko mice were far more susceptible to encephalitis. Delayed-type hypersensitivity to viral Ag was present, though diminished, in knockout mice, and serum herpes simplex virus-specific IgG isotypes indicated a Th2 shift. No differences existed in proliferative responses to in vitro Ag stimulation in gko vs control mice nor in T cell or proinflammatory cytokine mRNA levels in the corneas of infected mice. However, up-regulation of Th2 cytokine mRNA did occur in in vitro Ag-stimulated gko immune splenocytes. Histopathologic lesions were not statistically different between any of the groups of mice analyzed. These observations indicate that although IFN-gamma plays an important role in the clearance of virus from the eye, the pathogenesis of HSK lesions most likely involves additional cytokines, inflammatory mechanisms, or immune responses to nonviral Ags.


Modeling the time dependence of the association between human papillomavirus infection and cervical cancer precursor lesions.
The authors studied the time-dependent association between human papillomavirus (HPV) infection and squamous intraepithelial lesions (SIL) among women enrolled in a cohort study in Brazil (1993-2002), using repeated Papanicolaou cytologic examination and HPV testing by polymerase chain reaction. Through simulation with conceivable alternative cohort designs, they investigated different regression modeling approaches using time-varying covariates, time-varying hazard ratio functions, and repeated events to assess the effect of delay in lesion detection. Associations between HPV and early SIL were of high magnitude. The age-adjusted hazard ratios for the association between HPV at enrollment and low-grade SIL decreased gradually with time until 72 months for both oncogenic types of HPV (hazard ratio = 3.96, 95% confidence interval (CI): 2.5, 6.4) and nononcogenic types (hazard ratio = 2.37, 95% CI: 1.3, 4.3). The hazard ratio for incident high-grade SIL remained constant, ranging from 7.15 (95% CI: 2.0, 25.1) at 12 months to 6.26 (95% CI: 2.7, 14.5) at 72 months for oncogenic types of HPV. With oncogenic HPV as the time-dependent predictor variable, the hazard ratios for incident SIL and high-grade SIL events were 14.2 (95% CI: 8.7, 23.1) and 32.7 (95% CI: 8.4, 127.3), respectively. Investigators may underestimate the prognostic value of HPV detection using designs that rely on HPV ascertainment at a single time point. The waning in hazard ratios should be considered in the implementation of HPV testing-based screening programs.


Cerebrospinal fluid 5-hydroxytryptamine and 5-hydroxyindoleacetic acid in HIV-1 infection.
Reduced level of serotonin (5-hydroxytryptamine, 5-HT) in humans has been associated with a number of mental health and behavioral problems including depression, aggression, violence, sexual dysfunctions, sleep and eating disorders. Even though among HIV-1-infected individuals, prevalence of mental health and behavioral problems are common, their relationship with central nervous system serotonin functions is not clearly understood. This investigation was carried out to study the status of CSF 5-HT in HIV-1+ subjects (n = 21), in the early stage of infection, and HIV-1- control subjects (n = 24). Samples of CSF were obtained by lumbar puncture and were analyzed for 5-HT and its metabolite 5-hydroxyindoleacetic acid (5-HIAA), using high-performance liquid chromatography equipped with electrochemical detector. Levels of CSF 5-HT were significantly lower in the HIV-1+ group compared to the HIV-1- group. There was no significant difference in the CSF 5-HIAA levels between the two groups. In both groups, however, there was a significant correlation between CSF 5-HT and 5-HIAA. In the HIV-1 + group, although CSF 5-HT level was significantly negatively correlated with serostatus, there was no correlation between either CSF 5-HT or 5-HIAA levels and CD4 cell number or any behavioral measures evaluated in this study, including Beck's Depression Inventory and state/trait anxiety scores. These data suggest that HIV-1 infection affects the CNS 5-HT status with no significant association with measures of depression and anxiety, at least in the early stage of infection.


The dimeric structure of factor XI and zymogen activation.
Factor XI (fXI) is a homodimeric zymogen that is converted to a protease with 1 (1/2-fXIa) or 2 (fXIa) active subunits by factor XIIa (fXIIa) or thrombin. It has been proposed that the dimeric structure is required for normal fXI activation. Consistent with this premise, fXI monomers do not reconstitute fXI-deficient mice in a fXIIa-dependent thrombosis model. FXI activation by fXIIa or thrombin is a slow reaction that can be accelerated by polyanions. Phosphate polymers released from platelets (poly-P) can enhance fXI activation by thrombin and promote fXI autoactivation. Poly-P increased initial rates of fXI activation 30- and 3000-fold for fXIIa and thrombin, respectively. FXI monomers were activated more slowly than dimers by fXIIa in the presence of poly-P. However, this defect was not observed when thrombin was the activating protease, nor during fXI autoactivation. The data suggest that fXIIa and thrombin activate fXI by different mechanisms. FXIIa may activate fXI through a trans-activation mechanism in which the protease binds to 1 subunit of the dimer, while activating the other subunit. For activation by thrombin, or during autoactivation, the data support a cis-activation mechanism in which the activating protease binds to and activates the same fXI subunit.


Effects of endogenous steroids on CYP3A4-mediated drug metabolism by human liver microsomes.
In the present study, we investigated the effects of 14 endogenous steroids on the CYP3A4-mediated drug metabolism by human liver microsomes in vitro. Nevirapine (NVP) 2-, 12-hydroxylations, carbamazepine (CBZ) 10,11-epoxidation, triazolam (TZM) 1'-, 4-hydroxylations, erythromycin (EM) N-demethylation, and 2-sulphamoylacetylphenol (SMAP) formation from zonisamide (ZNS) were investigated. The activities of the NVP 2-, 12-hydroxylations, the CBZ 10,11-epoxidation, and the TZM 4-hydroxylation were activated by endogenous androgens, such as androstenedione (AND), testosterone, and dehydroepiandrosterone. However, these androgens inhibited EM N-demethylation, TZM 1'-hydroxylation, and SMAP formation. To understand the mechanisms of these effects of androgens on CYP3A4 activities, we performed a kinetic analysis of the metabolism of CBZ and ZNS in the presence or absence of AND using the modified two-site equation model. The addition of AND to the reaction mixture caused a drastic increase in the activity of CBZ 10,11-epoxidase, especially at a low substrate concentration, and resulted in a change in the kinetics from the sigmoid to Michaelis-Menten type. On the other hand, the metabolism of ZNS was strongly inhibited by AND, although no allosteric change was observed in this case. These data demonstrate that endogenous steroids, especially androgens, strongly affect CYP3A4-mediated drug metabolism in vitro. The postulated mechanisms of the interactions between AND and CBZ or ZNS are discussed.


Preconditioning by isoflurane retains its protection against ischemia-reperfusion injury in postinfarct remodeled rat hearts.
Postinfarct remodeling in the heart may affect protective signaling. We tested whether isoflurane preconditioning retains its protection in postinfarct remodeled hearts. Myocardial remodeling was induced by ligation of the left anterior descending coronary artery in male Wistar rats. Six weeks later, diseased hearts were mounted on a Langendorff apparatus and exposed to 40 min of ischemia followed by 90 min of reperfusion. Isoflurane preconditioning was induced with 1.5 MAC (2.1 vol%) isoflurane for 15 min. Infarct size was determined using 1% triphenyltetrazolium chloride staining and corroborated with measurements of lactate dehydrogenase (LDH) release into the perfusate. In some experiments, the protein kinase B and mitochondrial ATP-dependent potassium channel inhibitors LY294002 (10 microM) or 5-hydroxydecanoate (100 microM), respectively, were concomitantly added with isoflurane. Cardiac function was recorded. Six weeks after permanent coronary artery ligation, infarct rats exhibited a markedly increased heart weight/body weight index (5.41 +/- 0.64 vs 3.60 +/- 0.59 g/kg, P < 0.0001) confirming remodeling with compensatory hypertrophy. Isoflurane preconditioning decreased LDH release and reduced infarct size from 32% +/- 6% to 2% +/- 2% (P < 0.0001). Concomitant administration of LY294002 or 5-hydroxydecanoate with isoflurane completely abolished this protection. Functional assessment also showed significant protection from postischemic stunning by isoflurane preconditioning in remodeled hearts, which was lost in the presence of both blockers. Myocardial preconditioning with isoflurane retains its protection against ischemia/reperfusion injury in postinfarct remodeled rat hearts via similar signaling pathways, as previously reported in healthy hearts.


Protease inhibitors in the treatment of hereditary angioedema.
Deficiency of C1 Inhibitor leads to unopposed activation of complement, with localized, unpredictable, and sometimes life-threatening attacks of angioedema. Treatment with plasma-derived C1 Inhibitor rapidly aborts attacks, and may be lifesaving, but is expensive, requires use of a pooled blood product, may need to be repeated and may not be effective in autoantibody mediated angioedema. The antifibrinolytic agents aprotinin, tranexamic acid, and epsilon-aminocaproic acid are useful for prophylaxis and treatment of angioedema, likely by inhibiting plasmin. Specific drugs to replace the deficient C1 Inh have not been reported. The kallikrein inhibitor DX-88 (Dyax) has received orphan drug status in Europe and is undergoing clinical trial in Europe and the USA.


Fibroblast growth factor 2 uses distinct signaling pathways for cell proliferation and cell shape changes in corneal endothelial cells.
Fibroblast growth factor 2 (FGF-2) is not only a potent mitogen, it is a modulator for corneal endothelial cells. To define how the modulation activities of FGF-2 are mediated, we used pharmacologic inhibitors to examine the association of phospholipase C-gamma 1 (PLC-gamma) with FGF receptor or with cytoskeleton. Cell proliferation was determined either by the incorporation of 3H-thymidine into DNA or by counting cell numbers in the absence or presence of the inhibitors. The protein expression was analyzed by immunoprecipitation and immunoblot analysis. Cell shape change was determined by phase-contrast microscopy. FGF-2 stimulated DNA synthesis, whereas genistein inhibited the FGF-2-mediated cell proliferation in a dose-dependent manner, regardless of the concentration of FGF-2. The PLC-gamma 1 specific antisense oligonucleotide primer was able to inhibit cell proliferation by 25% in the absence of FGF-2; however, the antisense primer was not able to override the action of FGF-2. Fibroblast growth factor receptor was phosphorylated on treatment of the cells with FGF-2; however, 24-hour treatment with the growth factor significantly reduced phosphorylation of the receptor. Phospholipase C gamma 1 appears to be abundant in cytoplasm in the absence and presence of FGF-2, and a minor portion of the molecule is translocated to membrane after treatment with FGF-2; genistein inhibited the translocation. When the cytoskeleton fraction of the normal and the modulated corneal endothelial cells was immunoprecipitated with PLC-gamma 1 antibodies, PLC-gamma 1, actin, and vinculin were coprecipitated in both cell cultures. Phospholipase C gamma 1 associated with cytoskeleton was phosphorylated on treatment of the cells with FGF-2. In the presence of FGF-2 of the modulated cells, cytochalasin B, which did not revert the modulated cell morphology, abolished the association of PLC-gamma 1 with actin and vinculin; colchicine, which did revert the modulated cell shape to the polygonal shape, did not block the association of these three molecules. Interestingly, colchicine slightly enhanced the stimulatory effect of FGF-2 on corneal endothelial proliferation in contrast to the effect of cytochalasin B, which slightly decreased the FGF-2 action on cell proliferation. The association of PLC-gamma 1 with cytoskeleton plays a role in cell proliferation, whereas the association of PLC-gamma 1 with actin and vinculin has no effect on cell shape changes. These findings indicate that FGF-2 appears to use distinct signaling pathways for cell proliferation and cell shape changes in corneal endothelial cells.


Osteoporosis management in post-menopausal women.
Osteoporosis is most prevalent in women over the age of 50 as the hormonal influence of estrogen on bone health dissipates with the onset of menopause. The progressive changes in bone structure, quality and density lead to pathological fractures and an increase in morbidity and mortality among menopausal women. This review will examine the 2010 North American Menopause Society (NAMS) position statement and other recent publications to summarize the data and combinations of therapies used to treat women 50 years or older with osteoporosis. To review the latest research and guidelines for osteoporosis management we performed a PubMed search using the parameters Linked to free full text, Humans, Female, Review, English, Middle Age (45-64 years and 45+ years), Age 65+ years, and published in the last five years. Articles were sorted by relevance and hand searching of these articles was done to further increase the yield. While a perfect treatment has yet to be discovered to completely cure this progressive disease, many breakthroughs have been made in order to prevent fractures and improve quality of life. Calcium and vitamin D supplementation are recommended for patients undergoing pharmacological treatment, however, trials looking at their effectiveness have mixed findings. Bisphosphonates are considered the first line therapy in the treatment of osteoporosis and reduce vertebral fractures by 40% to 70% and non-vertebral fractures by 20% to 35%. Calcitonin showed promise during early trials in 2000 with a 33% reduction in fractures but these results have not been replicated and this therapy is now relegated to a second line treatment. Teriparatide is recommended for patients with severe osteoporosis and has been shown to reduce vertebral fractures 65% and non-vertebral fractures 53%. Selective estrogen receptor modulators (SERMs) are another useful therapy resulting in a 55% reduction in vertebral fractures without any documented advantage when looking at non-vertebral fractures. The currently available SERMs for this indication include raloxifene, available in the USA, and bazedoxifene, in Europe. Estrogen is effective, with a 27% reduction in fractures, but often is reserved for concomitant use for other menopausal symptoms or in patients intolerant of other available osteoporosis therapies. The newly approved monoclonal antibody for osteoporosis treatment in postmenopausal women, denosumab, leads to a 68% and 19% reduction of vertebral and non-vertebral fractures, respectively. In conclusion, the 2010 NAMS position statement provides an excellent framework to discuss treatment options with patients. Lifestyle optimization should be the bedrock of any good treatment approach. When pharmacological intervention is warranted, many good therapies are available which have been shown to reduce the risk of fractures in osteoporotic patients. Any treatment plan, however, will be ineffective if the patient is not compliant. Therefore, a detailed discussion regarding each therapeutic intervention should ensue, including its usefulness and side effects.


Investigation of de novo cholesterol synthetic capacity in the gonads of goldfish (Carassius auratus) exposed to the phytosterol beta-sitosterol.
Total and intra-mitochondrial gonadal cholesterol concentrations are decreased in fish exposed to the phytoestrogen beta-sitosterol (beta-sit). The present study examined the potential for beta-sit to disrupt de novo cholesterol synthesis in the gonads of goldfish exposed to 200 microgram/g beta-sit and 10 microgram/g 17beta-estradiol (E2; estrogenic control) by intra-peritoneal Silastic implants for 21 days. The de novo cholesterol synthetic capacity was estimated by incubating gonadal tissue with 14C-acetate for a period of 18 hours, followed by chloroform/methanol lipid extraction and thin layer chromatography (TLC) lipid separation. Lipid classes were confirmed using infrared spectroscopy. Plasma testosterone (T) and total cholesterol concentration were measured and gonadosomatic index (GSI) was calculated. Plasma T was significantly reduced in male beta-sit-treated fish compared to control and E2-treated fish (p < 0.001). 14C-Acetate incorporation into cholesterol and cholesterol esters was not significantly different among treatment groups for male and female fish, however, 14C-enrichment was higher than expected in both triglycerides (TG) and free fatty acids (FFA). FFA incorporation was significantly higher in male control fish than either beta-sit or E2 treatments (p = 0.005). Plasma cholesterol concentration was significantly increased in the male beta-sit treatment group compared to controls (p = 0.027). These results indicate gonadal de novo cholesterol biosynthetic capacity is not disrupted by beta-sit or E2 treatment in early recrudescing male or female goldfish, while plasma cholesterol and steroid concentrations are sensitive to beta-sit exposure.


Long-term outcome of major depressive disorder in psychiatric patients is variable.
The prevailing view of outcome of major depressive disorder (MDD), based on mostly inpatient cohorts sampled from tertiary centers, emphasizes chronicity and frequent recurrences. We investigated the long-term outcome of a regionally representative psychiatric MDD cohort comprising mainly outpatients. The Vantaa Depression Study included 163 patients with DSM-IV MDD (71.5% of those eligible) diagnosed using structured and semistructured interviews and followed up at 6 months, 18 months, and 5 years with a life chart between February 1, 1997, and April 30, 2004. The effects of comorbid disorders and other predictors on outcome were comprehensively investigated. Over the 5-year follow-up, 98.8% of patients achieved a symptom state below major depressive episode (MDE) criteria, and 88.4% reached full remission, with the median time to full remission being 11.0 months. Nearly one third (29.3%) had no recurrences, whereas 30.0% experienced 1, 12.9% experienced 2, and 27.9% experienced 3 or more recurrences. Preceding dysthymic disorder (p = .028), cluster C personality disorder (p = .041), and longer MDE duration prior to entry (p = .011) were the most significant predictors of longer time in achieving full remission. Severity of MDD and comorbidity, especially social phobia, predicted probability of, shorter time to, and number of recurrences. Previous literature on mostly inpatient MDD may have, by generalizing from patients with the most severe psychopathology, overemphasized chronicity of MDD. The long-term outcome of MDD in psychiatric care is variable, with about one tenth of patients having poor, one third having intermediate, and one half having favorable outcomes. In addition to known predictors, cluster C personality disorders and social phobia warrant further attention as predictors of MDD outcome among outpatients.


Spatial regulation of homeo box gene expression in Drosophila.
1. Each of the 17 known homeo box genes shows a unique pattern of expression during early embryonic development. Thus, virtually every embryonic cell contains a unique combination of active and inactive homeo box genes. 2. These different permutations of homeo box gene expression are thought to specify unique positional values, and appear to play key roles in selecting diverse pathways of morphogenesis. 3. It has been proposed that selective patterns of homeo box gene expression involve cross-regulatory interactions. For example, it appears that homeotic genes expressed in posterior regions of the embryo (such as abd-A and Abd-B) repress the expression of those homeotic genes expressed in more anterior regions (such as Antp and Ubx). Such interactions might involve sequence-specific DNA binding and occur at the level of transcription. 4. The establishment of selective patterns of pair-rule gene expression along the anterior-posterior embryonic body axis might involve the differential regulation of these genes by a common set of transcription factors. In particular, it appears that eve and ftz show opposite responses to regulatory products that are either directly or indirectly specified by several gap genes.


Zc3h10 is a novel mitochondrial regulator.
Mitochondria are the energy-generating hubs of the cell. In spite of considerable advances, our understanding of the factors that regulate the molecular circuits that govern mitochondrial function remains incomplete. Using a genome-wide functional screen, we identify the poorly characterized protein Zinc finger CCCH-type containing 10 (Zc3h10) as regulator of mitochondrial physiology. We show that Zc3h10 is upregulated during physiological mitochondriogenesis as it occurs during the differentiation of myoblasts into myotubes. Zc3h10 overexpression boosts mitochondrial function and promotes myoblast differentiation, while the depletion of Zc3h10 results in impaired myoblast differentiation, mitochondrial dysfunction, reduced expression of electron transport chain (ETC) subunits, and blunted TCA cycle flux. Notably, we have identified a loss-of-function mutation of Zc3h10 in humans (Tyr105 to Cys105) that is associated with increased body mass index, fat mass, fasting glucose, and triglycerides. Isolated peripheral blood mononuclear cells from individuals homozygotic for Cys105 display reduced oxygen consumption rate, diminished expression of some ETC subunits, and decreased levels of some TCA cycle metabolites, which all together derive in mitochondrial dysfunction. Taken together, our study identifies Zc3h10 as a novel mitochondrial regulator.