A test strip IgM dot-ELISA assay using leptospiral antigen of endemic strains for serodiagnosis of acute leptospirosis.
A test strip IgM dot-ELISA assay for the detection of leptospire-specific IgM antibodies in human sera was developed. Antigen dotted on a nitrocellulose paper strip was the pool sonicated antigen prepared from three predominant reactive Leptospira serovars currently in endemic area, i.e., Bratislava, Sejroe and Pyrogenes. The ability of the test to diagnose acute leptospiral infection was assessed by testing 343 single serum samples from 96 laboratory-confirmed leptospirosis case patients with positive result in the standard microscopic agglutination test (MAT), 55 serum samples from patients with various diseases other than leptospirosis, and 192 serum samples from healthy individuals. Using the results of the MAT as a gold standard, the sensitivity and specificity of the test strip IgM dot-ELISA assay were 98.96 and 93.93 per cent, respectively. The assay offered relatively high negative predictive values (99.57%) thus making the assay ideally suited for rapid screening. The stability of the test strip was assessed with a panel of five positive and five negative control sera after storage at 4 degrees C and -20 degrees C at different times. The results showed a good performance of the test strip at both storage temperatures for up to one year. In conclusion, the test strip IgM dot-ELISA assay was sufficiently sensitive for use as a screening test for serodiagnosis of acute leptospirosis. The assay was simple, inexpensive, and easy to perform for both a single test format and a large number of specimens. However, further studies are still needed to improve the stability of the test strip and assay reagents at ambient temperature, and to make the assay more rapidly and more user friendly.


Nutritional status, well-being and functional ability in frail elderly service flat residents.
To investigate nutritional status and its relationship to cognition, well-being, functional ability and energy intake in frail elderly service flat residents. Cross-sectional and prospective study. Two municipal service flat complexes. A total of 80 residents (median age 85.5 (79-90) y) with regular home care assistance participated. A subgroup of 35 residents took part in a re-examination 1 y later. Mini Nutritional Assessment (MNA), Short Portable Mental Status Questionnaire, Barthel Index and Health Index were used for the evaluation of nutritional, cognitive and ADL function and well-being, respectively. In all, 30% of the frail and chronically ill service flat residents were assessed as malnourished and 59% were at risk of malnutrition. The malnourished residents had worse cognitive conditions (P<0.001) and well-being (P<0.05), lower functional ability (P<0.01) and they had a greater need for daily assistance (P<0.05) than the other residents. The median night fast period was 14.0 (12.5-15.0) h. Five subjects classified as malnourished at baseline had lost a median of -9.6 kg (range -11.0 to +7.3 kg) (P<0.05) in body weight at the 1-y follow-up, which contrasted significantly from the weight stability in residents classified as at risk for malnutrition or well-nourished. Out of 10 residents, nine were assessed to have impending nutritional problems that related to impaired well-being, cognition, and functional ability. Malnourished residents had a significant weight loss over one year. Studies are needed to determine whether weight loss and nutrition-related dysfunction in service flat residents are preventable or treatable.


Genetic analysis of first-trimester miscarriages with a combination of cytogenetic karyotyping, microsatellite genotyping and arrayCGH.
Miscarriage is the spontaneous loss of an embryo or fetus before the 20th week of pregnancy. Most miscarriages occur before the end of the first trimester (<13 weeks). Although many risk factors relate to this occurrence, genetic factors play the most important role. Chromosomal abnormalities, including both numerical and structural anomalies, underlie the majority of miscarriages. In this study, we employed a comprehensive approach using cytogenetic karyotyping, polymerase chain reaction (PCR)-based genotyping, and microarray-based comparative genomic hybridization (arrayCGH) in combination to analyze chromosomal profiles of 115 first-trimester miscarriages of Chinese women. Seventy cases (61%) were found to have chromosomal anomalies, of which 90% were numerical and 10% were structural. Cytogenetic karyotyping identified 78.6% (55/70), PCR assays 2.9% (2 triploids), and arrayCGH 18.6% (13/70) of the anomalies. In this study, a microdeletion of 108 kb and four microduplications sizing from 300 to 1460 kb were observed. An advantage of using this combination approach is that microsatellite genotyping and arrayCGH can be accomplished in spite of culture failure and maternal cell contamination. In addition, arrayCGH can detect submicroscopic chromosomal anomalies and gene dosage alterations.


A Rapid Single-Cell Antimicrobial Susceptibility Testing Workflow for Bloodstream Infections.
Bloodstream infections are a significant cause of morbidity and mortality worldwide. The rapid initiation of effective antibiotic treatment is critical for patients with bloodstream infections. However, the diagnosis of bloodborne pathogens is largely complicated by the matrix effect of blood and the lengthy blood tube culture procedure. Here we report a culture-free workflow for the rapid isolation and enrichment of bacterial pathogens from whole blood for single-cell antimicrobial susceptibility testing (AST). A dextran sedimentation step reduces the concentration of blood cells by 4 orders of magnitude in 20-30 min while maintaining the effective concentration of bacteria in the sample. Red blood cell depletion facilitates the downstream centrifugation-based enrichment step at a sepsis-relevant bacteria concentration. The workflow is compatible with common antibiotic-resistant bacteria and does not influence the minimum inhibitory concentrations. By applying a microfluidic single-cell trapping device, we demonstrate the workflow for the rapid determination of bacterial infection and antimicrobial susceptibility testing at the single-cell level. The entire workflow from blood to categorical AST result can be completed in less than two hours.


Rapid Detection of Urinary Tract Infections via Bacterial Nuclease Activity.
Rapid and accurate bacterial detection methods are needed for clinical diagnostic, water, and food testing applications. The wide diversity of bacterial nucleases provides a rich source of enzymes that could be exploited as signal amplifying biomarkers to enable rapid, selective detection of bacterial species. With the exception of the use of micrococcal nuclease activity to detect Staphylococcus aureus, rapid methods that detect bacterial pathogens via their nuclease activities have not been developed. Here, we identify endonuclease I as a robust biomarker for E. coli and develop a rapid ultrasensitive assay that detects its activity. Comparison of nuclease activities of wild-type and nuclease-knockout E. coli clones revealed that endonuclease I is the predominant DNase in E. coli lysates. Endonuclease I is detectable by immunoblot and activity assays in uropathogenic E. coli strains. A rapid assay that detects endonuclease I activity in patient urine with an oligonucleotide probe exhibited substantially higher sensitivity for urinary tract infections than that reported for rapid urinalysis methods. The 3 hr turnaround time is much shorter than that of culture-based methods, thereby providing a means for expedited administration of appropriate antimicrobial therapy. We suggest this approach could address various unmet needs for rapid detection of E. coli.


Childhood thrombocytopenic purpura--review and a case report.
Childhood idiopathic thrombocytopenic purpura is a rare condition seen in children. A case of a 10 year old female child is reported. Bleeding from the gingiva and petechiae on the tongue were observed. The blood picture was normocytic normochromic with neutrophilia and thrombocytopenia. Bone marrow study was suggestive of marrow changes in idiopathic thrombocytopenic purpura. Dental extractions were postponed. Recognition of the disease by the dental profession is stressed.


A cyclophilin A CPR1 overexpression enhances stress acquisition in Saccharomyces cerevisiae.
Cyclophilins are conserved cis-trans peptidyl-prolyl isomerase that are implicated in protein folding and function as molecular chaperones. We found the expression of cyclophilin A, Cpr1, changes in response to exposure to yeast Saccharomyces cerevisiae to abiotic stress conditions. The effect of Cpr1 overexpression in stress responses was therefore examined. The CPR1 gene was cloned to the yeast expression vector pVTU260 under regulation of an endogenous alcohol dehydrogenase (ADH) promoter. The overexpression of Cpr1 drastically increased cell viability of yeast in the presence of stress inducers, such as cadmium, cobalt, copper, hydrogen peroxide, tert-butyl hydroperoxide (t-BOOH), and sodium dodecyl sulfate (SDS). The Cpr1 expression also enhanced the cell rescue program resulting in a variety of antioxidant enzymes including thioredoxin system (particularly, thioredoxin peroxidase), metabolic enzymes (glucose-6-phosphate dehydrogenase and glyceraldehyde-3-phosphate dehydrogenase), and molecular chaperones (Hsp104, Hsp90, Hsp60 and Hsp42). Thus, our study illustrates the importance of Cpr1 as a molecular chaperone that improves cellular stress responses through collaborative relationships with other proteins when yeast cells are exposed to adverse conditions, and it also premises the improvement of yeast strains.


A bayesian mixed regression based prediction of quantitative traits from molecular marker and gene expression data.
Both molecular marker and gene expression data were considered alone as well as jointly to serve as additive predictors for two pathogen-activity-phenotypes in real recombinant inbred lines of soybean. For unobserved phenotype prediction, we used a bayesian hierarchical regression modeling, where the number of possible predictors in the model was controlled by different selection strategies tested. Our initial findings were submitted for DREAM5 (the 5th Dialogue on Reverse Engineering Assessment and Methods challenge) and were judged to be the best in sub-challenge B3 wherein both functional genomic and genetic data were used to predict the phenotypes. In this work we further improve upon this previous work by considering various predictor selection strategies and cross-validation was used to measure accuracy of in-data and out-data predictions. The results from various model choices indicate that for this data use of both data types (namely functional genomic and genetic) simultaneously improves out-data prediction accuracy. Adequate goodness-of-fit can be easily achieved with more complex models for both phenotypes, since the number of potential predictors is large and the sample size is not small. We also further studied gene-set enrichment (for continuous phenotype) in the biological process in question and chromosomal enrichment of the gene set. The methodological contribution of this paper is in exploration of variable selection techniques to alleviate the problem of over-fitting. Different strategies based on the nature of covariates were explored and all methods were implemented under the bayesian hierarchical modeling framework with indicator-based covariate selection. All the models based in careful variable selection procedure were found to produce significant results based on permutation test.


A predictive model to allocate frequent service users of community-based mental health services to different packages of care.
To develop predictive models to allocate patients into frequent and low service users groups within the Italian Community-based Mental Health Services (CMHSs). To allocate frequent users to different packages of care, identifying the costs of these packages. Socio-demographic and clinical data and GAF scores at baseline were collected for 1250 users attending five CMHSs. All psychiatric contacts made by these patients during six months were recorded. A logistic regression identified frequent service users predictive variables. Multinomial logistic regression identified variables able to predict the most appropriate package of care. A cost function was utilised to estimate costs. Frequent service users were 49%, using nearly 90% of all contacts. The model classified correctly 80% of users in the frequent and low users groups. Three packages of care were identified: Basic Community Treatment (4,133 Euro per six months); Intensive Community Treatment (6,180 Euro) and Rehabilitative Community Treatment (11,984 Euro) for 83%, 6% and 11% of frequent service users respectively. The model was found to be accurate for 85% of users. It is possible to develop predictive models to identify frequent service users and to assign them to pre-defined packages of care, and to use these models to inform the funding of psychiatric care.


Surface plasmon resonance in CdSe semiconductor coated with gold nanoparticles.
We have grown CdSe semiconductor films on glass substrates and the films were coated with Au nanoparticles of 10 nm in size by the pulsed-laser deposition technique. The films demonstrate a large enhancement of Raman intensity and photoluminescence of CdSe semiconductor via excitation of surface plasmon resonances in proximate gold metal nanoparticles deposited on the surface of CdSe film. These observations suggest a variety of approaches for improving the performance of devices such as photodetectors, photovoltaics, and related devices, including biosensors.


[Ontogenic differences of glutamate decarboxylase, GABA-aminotransferase, monoamine oxidase activities of cerebral cortex mitochondria and hypothalamic cerebral area].
It is shown that the glutamate decarboxylase activity in the initial mitochondrial fraction of "light" and "heavy" synaptosomes of cerebral hemispheres changes on the 14th, 21st day, 1st, 3d, 12th and 24th months of rat development: in the hypothalamus area it lowers in the initial mitochondria in the 21-day animals, and in the fraction of "heavy" synaptosomes--in the 24 day animals as compared to the adult ones. The GABA-aminotransferase activity in the cortex synaptosome fraction rises in the period between the 14th and 21st day after birthday and in the initial mitochondrial fraction of the hypothalamus area--between the 14th day and the first month of life. In adult rats the glutamate decarboxylase and GABA-aminotransferase activities in the "heavy" synaptosome fraction are higher than in the "light" synaptosome one. The amine oxidase activity of the subcellular fractions which is determined with serotonin as a substrate (form A) is stable during the whole developmental period of studies. The intensity of benzylamine (amine oxidase B) desamination in the cortex fraction and hypothalamus area rises sharply in the second half of ontogenesis.


Latent Regression Analysis.
Finite mixture models have come to play a very prominent role in modelling data. The finite mixture model is predicated on the assumption that distinct latent groups exist in the population. The finite mixture model therefore is based on a categorical latent variable that distinguishes the different groups. Often in practice distinct sub-populations do not actually exist. For example, disease severity (e.g. depression) may vary continuously and therefore, a distinction of diseased and not-diseased may not be based on the existence of distinct sub-populations. Thus, what is needed is a generalization of the finite mixture's discrete latent predictor to a continuous latent predictor. We cast the finite mixture model as a regression model with a latent Bernoulli predictor. A latent regression model is proposed by replacing the discrete Bernoulli predictor by a continuous latent predictor with a beta distribution. Motivation for the latent regression model arises from applications where distinct latent classes do not exist, but instead individuals vary according to a continuous latent variable. The shapes of the beta density are very flexible and can approximate the discrete Bernoulli distribution. Examples and a simulation are provided to illustrate the latent regression model. In particular, the latent regression model is used to model placebo effect among drug treated subjects in a depression study.


Automated sub-cortical brain structure segmentation combining spatial and deep convolutional features.
Sub-cortical brain structure segmentation in Magnetic Resonance Images (MRI) has attracted the interest of the research community for a long time as morphological changes in these structures are related to different neurodegenerative disorders. However, manual segmentation of these structures can be tedious and prone to variability, highlighting the need for robust automated segmentation methods. In this paper, we present a novel convolutional neural network based approach for accurate segmentation of the sub-cortical brain structures that combines both convolutional and prior spatial features for improving the segmentation accuracy. In order to increase the accuracy of the automated segmentation, we propose to train the network using a restricted sample selection to force the network to learn the most difficult parts of the structures. We evaluate the accuracy of the proposed method on the public MICCAI 2012 challenge and IBSR 18 datasets, comparing it with different traditional and deep learning state-of-the-art methods. On the MICCAI 2012 dataset, our method shows an excellent performance comparable to the best participant strategy on the challenge, while performing significantly better than state-of-the-art techniques such as FreeSurfer and FIRST. On the IBSR 18 dataset, our method also exhibits a significant increase in the performance with respect to not only FreeSurfer and FIRST, but also comparable or better results than other recent deep learning approaches. Moreover, our experiments show that both the addition of the spatial priors and the restricted sampling strategy have a significant effect on the accuracy of the proposed method. In order to encourage the reproducibility and the use of the proposed method, a public version of our approach is available to download for the neuroimaging community.


Thermal Simulation and Experimental Analysis of Optically Pumped InP-on-Si Micro- and Nanocavity Lasers.
There is a general trend of downscaling laser cavities, but with high integration and energy densities of nanocavity lasers, significant thermal issues affect their operation. The complexity of geometrical parameters and the various materials involved hinder the extraction of clear design guidelines and operation strategies. Here, we present a systematic thermal analysis of InP-on-Si micro- and nanocavity lasers based on steady-state and transient thermal simulations and experimental analysis. In particular, we investigate the use of metal cavities for improving the thermal properties of InP-on-Si micro- and nanocavity lasers. Heating of lasers is studied by using Raman thermometry and the results agree well with simulation results, both revealing a temperature reduction of hundreds of kelvins for the metal-clad cavity. Transient simulations are carried out to improve our understanding of the dynamic temperature variation under pulsed and continuous wave pumping conditions. The results show that the presence of a metal cladding not only increases the overall efficiency in heat dissipation but also causes a much faster temperature response. Together with optical experimental results under pulsed pumping, we conclude that a pulse width of 10 ns and a repetition rate of 100 kHz is the optimal pumping condition for a 2 μm wide square cavity.


Implementation of a multidisciplinary robotic centre in a high-volume university hospital.
We describe the initiation of a multidisciplinary centre for robotic surgery including the implementation of robotic-assisted procedures as standard procedure for the majority of cancer operations in urology, gynaecology and gastrointestinal surgery. All robotic procedures performed from 2008 to 2013 were included. The information gathered included body mass index, the American Society of Anesthesiologists' physical status classification value (ASA), age, sex, time and type of surgery, duration of procedure, conversion to open surgery, length and type of anaesthesia, re-operations, length of hospital stay and 30-day mortality. The implementation strategy was to start with one specialty at a time, passing on experience from one specialty to the next. The surgical strategy was to begin with standard procedures for which international experience was available and subsequently perform more complex procedures, ending up with robotic-assisted procedures as the standard for most cancer surgery procedures. A total of 2,473 procedures were performed. The operative time was reduced over the period for the main procedures of all three specialties. For prostatectomies, hysterectomies and colectomies, conversion to open surgery occurred in 1.2, 3.8 and 7.7%; the risk of re-operation was 0.2, 2.3 and 7.3%; and, finally, the 30-day mortality was 0.1, 0 and 1%, respectively. The implementation was possible as a stepwise introduction across three specialties with low conversion and re-operation rates and a low mortality. A high-volume centre for robotic surgery was developed and patients with malignant diagnoses were offered robotic-assisted surgery within the framework of multidisciplinary cooperation. not relevant. The study was approved by the Danish Data Protection Agency R. No.: 2007-58-0015.


[Role of cytoplasmic receptors in the lympholytic action of glucocorticoids and ionizing radiation].
Exposure of thymocytes to 4 Gy X-radiation causes a marked change in the kinetics of accumulation of glucocorticoids in cells. The hormone-binding activity of the cytoplasmic receptors does not differ from the control level. Progesterone, a glucocorticoid antagonist, and aurin tricarboxylic acid, an inhibitor of protein receptors, protect thymocytes from the effect of glucocorticoids and ionizing radiation. It is assumed that the cytoplasmic receptor system is involved in the realization of the radiation effect.


Serum Trypsinogen Levels in Type 1 Diabetes.
The pancreas in type 1 diabetes exhibits decreased size (weight/volume) and abnormal exocrine morphology. Serum trypsinogen levels are an established marker of pancreatic exocrine function. As such, we hypothesized that trypsinogen levels may be reduced in patients with pre-type 1 diabetes and type 1 diabetes compared with healthy control subjects. Serum trypsinogen levels were determined in 100 persons with type 1 diabetes (72 new-onset, 28 established), 99 autoantibody-positive (AAb+) subjects at varying levels of risk for developing this disease, 87 AAb-negative (AAb-) control subjects, 91 AAb- relatives with type 1 diabetes, and 18 patients with type 2 diabetes. Trypsinogen levels increased significantly with age in control subjects (r = 0.71; P < 0.0001) and were significantly lower in patients with new-onset (mean ± SD 14.5 ± 6.1 ng/mL; P < 0.0001) and established type 1 diabetes (16.7 ± 6.9 ng/mL; P < 0.05) versus AAb- control subjects (25.3 ± 11.2 ng/mL), AAb- relatives (29.3 ± 15.0 ng/mL), AAb+ subjects (26.5 ± 12.1 ng/mL), and patients with type 2 diabetes (31.5 ± 17.3 ng/mL). Multivariate analysis revealed reduced trypsinogen in multiple-AAb+ subjects (P < 0.05) and patients with type 1 diabetes (P < 0.0001) compared with AAb- subjects (control subjects and relatives combined) and single-AAb+ (P < 0.01) subjects when considering age and BMI. These findings further support the interplay between pancreatic endocrine and exocrine dysfunction. Longitudinal studies are warranted to validate trypsinogen as a predictive biomarker of type 1 diabetes progression.


Calculation of change in brain temperatures due to exposure to a mobile phone.
In this study we evaluated for a realistic head model the 3D temperature rise induced by a mobile phone. This was done numerically with the consecutive use of an FDTD model to predict the absorbed electromagnetic power distribution, and a thermal model describing bioheat transfer both by conduction and by blood flow. We calculated a maximum rise in brain temperature of 0.11 degrees C for an antenna with an average emitted power of 0.25 W, the maximum value in common mobile phones, and indefinite exposure. Maximum temperature rise is at the skin. The power distributions were characterized by a maximum averaged SAR over an arbitrarily shaped 10 g volume of approximately 1.6 W kg(-1). Although these power distributions are not in compliance with all proposed safety standards, temperature rises are far too small to have lasting effects. We verified our simulations by measuring the skin temperature rise experimentally. Our simulation method can be instrumental in further development of safety standards.


[Immediate and early results of combined treatment of colonic cancer using neoadjuvant selective intra-arterial polychemotherapy].
The combined treatment of 53 patients with colonic cancer, using neoadjuvant intraarterial polychemotherapy (IAPCT), was conducted. The trustworthy lowering of the catheterized artery thrombosis occurrence rate, the tumor recurrence-by 7%, distant metastases-by 13.9%, two-year survival increase-by 15.5% was promoted by verapamil application as a IAPCT modifier.


Randomized trial of a physician-based intervention to increase the use of folic acid supplements among women.
Fewer than one third of American women take folic acid daily, although many women report that they would take folic acid if their physicians advised them to do so. This study determined the impact of a physician intervention during routine gynecologic visits on folic acid supplementation. Patients were assigned randomly to receive brief folic acid counseling, a reminder phone call, and 30 folic acid tablets (n = 162 women; intervention group) or to receive counseling about other preventive health behaviors and a folic acid informational pamphlet (n = 160 women; control group). Self-reported folic acid use was compared at baseline and at 2 months. Of the 279 patients who completed the study, weekly folic acid intake increased in the intervention group by 68%, compared with 20% in the control group ( P =.008). No significant differences were found in daily intake. The women who were most influenced by the intervention were black and lower income and not planning pregnancies. With little effort expended to encourage folic acid use, gynecologists could potentially reduce the risk of folate-preventable birth defects among their patients by as much as 11%.


Ambivalent Adherence and Nonadherence to Continuous Positive Airway Pressure Devices: A Qualitative Study.
Continuous positive airway pressure (CPAP) therapy is considered the front-line treatment for moderate-severe obstructive sleep apnea (OSA). However, nonuse rates are very high, such that adherence to CPAP has become a major concern. Although the literature on CPAP use is vast, further research is required to understand patients' experiences of CPAP use and nonuse. This is the goal of this study. This study draws on in-depth interviews with 61 Jewish-Israeli patients with OSA who received a recommendation to use a CPAP device. The sample includes both patients who started using CPAP devices as well as patients who rejected this course of treatment. It follows principles of constructivist-grounded theory in both sampling and analysis. The study shows that regardless of patients' status of adherence, their attitudes toward CPAP devices are characterized by ambivalence. Users of CPAP expressed ambivalent adherence, pondering whether they should stop using the device; and patients who rejected the CPAP expressed ambivalent nonadherence, wondering whether they should give the CPAP another chance. This study identifies the experiences involved in using, as well as not using, CPAP devices that produce patients' ambivalence. Both adherence and nonadherence to CPAP are dynamic processes that are characterized by patients' ambivalence and contingent upon diverse factors. These findings have practical implications as they suggest that all patients, regardless of their initial adherence status, would benefit from a close follow-up.


Nursing and the Public Health Legacies of the Russian Revolution.
The centenary of the October Revolution in 1917 provides a timely opportunity to assess the legacies of that event. I examine the role of the revolution in public health with a focus on nursing, assessing the Imperial Russian health care system, the development of Soviet nursing, and current plans for nursing and public health care in Putin's Russia. Analyzing nursing shows that there was a great deal of continuity in terms of medical personnel and ideas on how public health care service in Russia should operate. Nursing illuminates some of the complexities of Soviet health care and ideology, particularly the state's desire to create a socialist form of nursing in theory, despite the strong links with the prerevolutionary past in the form of personnel. This situation changed after the collapse of the Soviet Union, when the new Russian state attempted to sever connections with the past, this time with the Soviet past. But as I show, making a clean break with the past is a difficult and often fraught process.


Pharmacological profiles and clinical effects of azelnidipine, a long-acting calcium channel blocker.
Azelnidipine (Calblock) is a newly developed dihydropyridine-type calcium antagonist for the treatment of hypertension. In hypertensive animals, a single oral administration of azelnidipine caused a slowly developed and long-lasting hypotensive effect with a little reflex tachycardia. The extent of tachycardia was less with azelnidipine than with other agents of the same class. Long-term administrations of azelnidipine produced a stable antihypertensive effect with a slight decrease in heart rate. The hypotensive effect was preceded by an increase in plasma drug concentration and it persisted even after plasma drug concentration declined to very low levels. In the isolated arteries, the calcium blocking action developed gradually after treatment with azelnidipine and survived for a long period of time after the drug was removed from the bathing solution. These data suggest that the high affinity to vascular tissue contributes to the long-lasting hypotensive effects of this agent. The results from clinical studies in hypertensive patients indicated that once daily administration of azelnidipine achieved stable, 24-h control of blood pressure with no change or a slight decrease in heart rate. Clinical studies also showed a low incidence of adverse events such as headache, facial flush, dizziness, and palpitations. These characteristics make azelnidipine a new generation calcium antagonist that can be used for the treatment of hypertension.


Combined optical fluorescence microscopy and X-ray tomography reveals substructures in cell nuclei in 3D.
The function of a biological cell is fundamentally defined by the structural architecture of packaged DNA in the nucleus. Elucidating information about the packaged DNA is facilitated by high-resolution imaging. Here, we combine and correlate hard X-ray propagation-based phase contrast tomography and visible light confocal microscopy in three dimensions to probe DNA in whole cell nuclei of NIH-3T3 fibroblasts. In this way, unlabeled and fluorescently labeled substructures within the cell are visualized in a complementary manner. Our approach enables the quantification of the electron density, volume and optical fluorescence intensity of nuclear material. By joining all of this information, we are able to spatially localize and physically characterize both active and inactive heterochromatin, euchromatin, pericentric heterochromatin foci and nucleoli.


[Primary squamous cell carcinoma of the ascending colon: report of a case and Korean literature review].
Primary squamous cell carcinoma of the colon is an extremely rare malignancy. A 48-year-old male visited our hospital for screening colonoscopy. Colonoscopic examination showed a 1 cm sized sessile polyp in the ascending colon. The patient underwent endoscopic mucosal resection (EMR) without any complication. The pathologic findings were compatible with squamous differentiation of tumor cells in inflammatory colonic mucosa. The tumor was confined to the mucosa and the margins of the excised tissue were found to be free of the tumor. There were no other primary sites and no distant metastases in the extensive evaluation using a whole body CT scan and PET-CT. Additional surgical resection was not done. Follow-up colonoscopy performed eight month later showed a whitish scar without evidence of local recurrence and follow-up PET-CT demonstrated no evidence of recurrence. Herein, we report a case of primary squamous cell carcinoma of the ascending colon presenting as a sessile polyp which was removed by EMR.


Women Coping With a Partner's Dementia-Related Violence: A Qualitative Study.
The aim of the present study was to differentiate between the lived experience of two groups of women caregiving for a partner with dementia. One group was coping with lifelong intimate partner violence (IPV) and dementia-related violence (Group 1); the other group was coping with dementia-related violence only (Group 2). An interpretive phenomenological analysis perspective was used. Data collection was performed through in-depth, semistructured interviews with eight female spouses of men with dementia from each of the two above-mentioned groups, followed by a content analysis. Comparing the narratives of the aging women revealed their different experiences over several dimensions: (a) the identification of violence as a symptom of dementia; (b) the use of past couplehood memories; (c) feelings over time; (d) willingness to care for the partner with dementia; and (e) a prospective view of life. The complexities of being old and having to cope with caregiving responsibilities for a spouse with dementia, accompanied by violent behaviors, emphasize the significance of the couple's past relationship. This notion should be taken into account in practical interventions. As part of the aging process, there is an increase in people who are engaged in dementia-related violence. Nurses' education and practice should focus on the dynamics of dyads coping with violence and identify the particular needs of the caregiver spouse in this context.


An aminoglycoside antibiotic gentamycin induces oxidative stress, reduces antioxidant reserve and impairs spermatogenesis in rats.
Gentamycin (GS) is an aminoglycoside antibiotic used to treat infections of various Gram-negative organisms. The present study was designed to investigate the effects of GS on oxidative stress, antioxidant levels, testicular structure and sperm parameters in the rat. Adult Wistar rats (12 week old; N=7/group) were treated (i. p.) with 0 mg/kg, 3 mg/kg and 5 mg/kg for 10 days at an interval of 24 hr between subsequent treatments. Animals were sacrificed on days 1 and 35 after the last treatment, and the reproductive organs were removed and weights of testis and seminal vesicle were recorded. The right testis was processed for light microscopic analysis. The left testis was homogenized and step 19 spermatids were counted to determine the daily sperm production (DSP) and daily abnormal sperm production (DASP). The sperm count, sperm motility and incidence of abnormal sperms were estimated in the epididymis. In testicular sections, along with the evaluation of qualitative changes, the seminiferous tubule diameter (STD) and the epithelial height (SE) were measured. The incidence of stage XIV tubules in testicular sections, meiotic figures and step 14 spermatids/stage XIV tubule, and step 19 spermatids/stage VII tubule were estimated. Intra-testicular levels of superoxide anion, lipid peroxidation and antioxidants-superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx) and ascorbic acid were measured. GS did not affect the body weight, but the testis weight and DSP were decreased at 5 mg/kg dose-level on both days (p<0.05), and the weight of seminal vesicle decreased on day 35 at both dose-levels. The DASP was increased in a dose-dependent manner (p<0.05) on days 1 and 35 at both dose-levels. The sperm count was decreased at both dose-levels on day 35, whereas the sperm motility was decreased and sperm abnormality was increased on day 1 at 5 mg/kg and on day 35 at both dose-levels. GS induced structural changes such as sloughing of seminiferous epithelium, vacuoles and gaps in the epithelium, nuclear pyknosis and atrophic changes in a few tubules. The tubular shrinkage was observed as indicated by decreased STD and SE on both days at 5 mg/kg dose-level. Incidence of stage XIV tubules and step 19 spermatids/stage VII tubule decreased on all time points at all dose-levels, whereas the step 14 spermatids and meiotic figures decreased on day 35 at both dose-levels (p<0.05). The free radical- superoxide anion concentration was significantly increased on day 1 in a dose-dependent pattern (p<0.05). However, activities of all 3 enzymatic antioxidants and ascorbic acid level decreased in a dose-dependent pattern on day 1 (p<0.05), except the GPx, which was also decreased on day 35 at 5 mg/kg dose-level. There was a significant rise in the thiobarbituric acid reactive substances on day 1 indicating increased lipid peroxidation in the testis. In conclusion, GS induces an oxidative stress-status in the testis by increasing free radical formation and lipid peroxidation, and by decreasing the antioxidant reserves. These biochemical changes manifest as structural and cytotoxic changes in the testis. Further, GS also affects the spermatozoa by affecting their number, motility and morphology.


Parkin mediates the degradation-independent ubiquitination of Hsp70.
Mutations in the parkin gene cause autosomal recessive, juvenile-onset parkinsonism. Parkin is an E3 ubiquitin ligase that mediates the ubiquitination of protein substrates. Disease-associated mutations cause a loss-of-function of parkin which may compromise the poly-ubiquitination and proteasomal degradation of specific protein substrates, potentially leading to their deleterious accumulation. Here, we identify the molecular chaperones, Hsp70 and Hsc70, as substrates for parkin. Parkin mediates the ubiquitination of Hsp70 both in vitro and in cultured cells. Parkin interacts with Hsp70 via its second RING finger domain and mutations in/near this domain compromise Hsp70 ubiquitination. Ubiquitination of Hsp70 fails to alter its steady-state levels or turnover, nor does it promote its proteasomal degradation. Consistent with this observation, Hsp70 levels remain unaltered in brains from parkin-deficient autosomal recessive, juvenile-onset parkinsonism subjects, whereas alternatively, Hsp70 levels are elevated in the detergent-insoluble fraction of sporadic Parkinson's disease/dementia with Lewy bodies brains. Parkin mediates the multiple mono-ubiquitination of Hsp70/Hsc70 consistent with a degradation-independent role for this ubiquitin modification. Our observations support a novel functional relationship between parkin and Hsc/Hsp70 and support the notion that parkin is a multi-purpose E3 ubiquitin ligase capable of modifying proteins either via attachment of alternatively linked poly-ubiquitin chains or through multiple mono-ubiquitination to achieve alternate biological outcomes.


Candida albicans Zn Cluster Transcription Factors Tac1 and Znc1 Are Activated by Farnesol To Upregulate a Transcriptional Program Including the Multidrug Efflux Pump CDR1.
Farnesol, a quorum-sensing molecule, inhibits Candida albicans hyphal formation, affects its biofilm formation and dispersal, and impacts its stress response. Several aspects of farnesol's mechanism of action remain incompletely uncharacterized. Among these are a thorough accounting of the cellular receptors and transporters for farnesol. This work suggests these processes are linked through the Zn cluster transcription factors Tac1 and Znc1 and their induction of the multidrug efflux pump Cdr1. Specifically, we have demonstrated that Tac1 and Znc1 are functionally activated by farnesol through a mechanism that mimics other means of hyperactivation of Zn cluster transcription factors. This is consistent with our observation that many genes acutely induced by farnesol are dependent on TAC1, ZNC1, or both. A related molecule, 1-dodecanol, invokes a similar TAC1-ZNC1 response, while several other proposed C. albicans quorum-sensing molecules do not. Tac1 and Znc1 both bind to and upregulate the CDR1 promoter in response to farnesol. Differences in inducer and DNA binding specificity lead to Tac1 and Znc1 having overlapping, but nonidentical, regulons. Induction of genes by farnesol via Tac1 and Znc1 was inversely related to the level of CDR1 present in the cell, suggesting a model in which induction of CDR1 by Tac1 and Znc1 leads to an increase in farnesol efflux. Consistent with this premise, our results show that CDR1 expression, and its regulation by TAC1 and ZNC1, facilitates growth in the presence of high farnesol concentrations in C. albicans and in certain strains of its close relative, C. dubliniensis.


Distribution and environmental risk evaluation of heavy metal in core sediments from Lake Çıldır (NE Turkey).
This study examined the vertical distribution of heavy metals in core sediments taken from Lake Çıldır, Turkey, and their potential ecological risks together with organic carbon content and chlorophyll degradation products. Samples were collected from six sampling stations determined along two main transections aligned in north-south and east-west directions. The enrichment (EF) and contamination factor (CF), potential ecological risk (PER) index, and pollution load index (PLI) were calculated from the obtained results. For the elements Pb, As, and Cd, a moderate level of contamination was detected, whereas a moderate-to-high concentration level was obtained for Mn. The highest contamination level was found for Hg. A pollutant accumulation exists particularly in the surface sediments. Cd and Hg are the only two metals considered to be a potential risk factor in the lake.


Assessing risk for pelvic inflammatory disease and its sequelae.
To assess the risk for pelvic inflammatory disease (PID), a practitioner must evaluate the likelihood that a woman has PID or will be exposed to a sexually transmitted disease causing PID. Successful risk assessment depends on accurate information about variables influencing risk of PID. To determine the current state of knowledge about PID risk variables, we examined data in published reports. Data on each risk variable were scrutinized to discern which link(s) in the PID risk chain it affects (acquisition of a sexually transmitted disease, development of PID, or development of PID sequelae) and whether it is a risk marker or a risk factor. Most PID risk variables, particularly sexual behaviors, are associated with acquisition of a sexually transmitted disease, rather than development of PID itself. With the exception of age, demographic and social indicators of risk appear to be risk markers, while contraceptive practices appear more often to be risk factors than risk markers. Additional data are needed for most PID risk variables confidently to categorize them as risk factors. Enough information is available, however, to begin assessing risk for PID, so that appropriate counseling can ensue and timely diagnosis can be made.


Ischaemic stroke and bleeding rates in 'real-world' atrial fibrillation patients.
Stroke prevention guidelines recommend oral anticoagulants (OAC) for atrial fibrillation (AF) patients at moderate/high risk of stroke, and antiplatelet or no therapy for those at low/moderate risk. Outcomes for AF patients receiving antiplatelet/no therapy in 'real-life' clinical practice were explored. This study compared clinical event rates (stroke/bleeding) for AF patients treated with OAC therapy, antiplatelets or no therapy in usual clinical practice to event rates in OAC-treated AF patients from optimally-monitored 'real-life' settings (anticoagulation clinics). We searched biomedical literature (1994-2010) using PubMed to identify 'real-world' studies of clinical event rates for AF patients receiving OAC therapy, antiplatelets, or no therapy; event rates were extracted for each treatment and setting. We identified 136 studies of thromboembolic events and 86 of bleeding events. Ischaemic stroke rates (30 studies) were higher for AF patients receiving no therapy (median: 4.45/100 person-years; range: 0.25-5.9) or antiplatelet-therapy (median: 4.45/100 person-years; range: 2.0-10) compared to OAC-treated patients monitored in anticoagulation clinics (median: 1.72/100 person-years; range: 0.97-2.00), or from a non-specialized setting (median 1.66/100 person-years; range: 0-4.9). Major bleeding rates (32 studies) for patients receiving antiplatelet/no therapy were similar to OAC-treated patients from both clinical settings. As in randomised clinical trials, AF patients in 'real-world' clinical practice receiving antiplatelet/no therapy have higher rates of ischaemic stroke than OAC-treated patients. Antiplatelet/no therapy was associated with similar bleeding rates to OAC therapy. Increasing utilisation of anticoagulants in clinical practice could improve patient outcomes.


Sink or swim--clinicians don't often counsel on drowning prevention.
Drowning is one of the leading causes of injury death for young children in the United States. This study examined primary care providers' knowledge of and counseling on drowning prevention. A random sample of 465 Los Angeles County pediatricians, family physicians, and pediatric nurse practitioners who serve families with young children received mailed questionnaires; 325 (70%) responded. About two thirds of clinicians did not know that injury deaths attributable to drowning were more common than those attributable to toxic ingestions and firearm injuries in young children. Only one third of clinicians stated they counseled on drowning prevention. Counseling drowning prevention was positively associated with female gender (odds ratio: 1.97; 95% confidence interval: 1.64, 2.30) and negatively associated with an attitude that drowning prevention counseling was less important than other injury prevention topics (odds ratio:.73; 95% confidence interval:.61,.85). Clinician specialty, age, years out from training, proportion of well-child examinations in a typical week, having children, practice setting, and knowledge of drowning injury deaths were not significant in multivariate analysis. The belief of clinicians that it is less important to counsel on drowning prevention than other injury prevention topics poses a substantial challenge to their providing such education to families with young children.drowning, counseling, physician's role.


Short communication: Identification and differentiation of bifidobacteria obtained from Ukraine.
Ten freeze-dried bifidobacterial strains used as probiotics in Ukrainian dairy foods, identified by the supplier as Bifidobacterium adolescentis (2), Bifidobacterium bifidum (2), Bifidobacterium longum (4), Bifidobacterium animalis (1), and Bifidobacterium infantis (1), were characterized. Following rehydration and anaerobic growth on de Man, Rogosa, and Sharpe-cysteine medium at 37°C for 72 h, single-colony isolates were picked and evaluated using PCR primers specific for the Bifidobacterium genus, for the supplier-identified species, and for B. animalis ssp. lactis. All isolates were identified as members of the genus Bifidobacterium; however, species-specific PCR revealed all 10 isolates were actually strains of B. animalis ssp. lactis. Further evaluation using pulsed-field gel electrophoresis was only able to separate a single strain (RT 09) from the other 9 strains evaluated. Application of genome-wide allelic profiling to the Ukrainian bifidobacterial strains revealed 4 distinct groups. Interestingly, 6 (60%) of the isolates fell into the same cluster as that containing the common commercial probiotic strain BB-12. Two of the strains (RT 02 and RT 09) were found to be in the same group as ATCC 27536 and one strain (RT 08) was in the same group as the RB 7239 (a previously evaluated commercial strain). One strain, RT 04, was placed on a unique branch. These results highlight the importance of employing routine typing of bifidobacterial isolates, demonstrate the utility of single nucleotide polymorphism/insertion-deletion polymorphism-based allelic typing in B. animalis ssp. lactis strain differentiation and further point to the limited genetic variability of B. animalis ssp. lactis strains and the worldwide distribution of a small number of commercial strains.


Optical imaging of retina in response to grating stimuli in cats.
We examined the intrinsic signals in response to grating stimuli in order to determine whether the light-evoked intrinsic signals of the retina are due to changes in the photoreceptor activities induced by the image projected on to the retina or are due to neural activities of the inner retina. The retinas of the left eye of 12 cats under general anesthesia were examined by a functional imaging fundus camera. Near infrared light was used to monitor the reflectance changes (RCs) of the retina. Vertical grating were used to stimulate the retina at 4 Hz. The spatial frequencies of the gratings were 0.05, 0.11, 0.22, 0.43, 0.86, 1.73, and 3.46 cycles/degree (cpd). Ten images were averaged and used to analyze the RCs to obtain the peak value (PV) of a two dimensional fast Fourier transfer of the RCs. The wavefront aberrations (WA) were measured with a compact wavefront aberrometer and the spatial modulation transfer function (MTF) of the eye was calculated. The retinal reflectance image had a grating pattern. The PV of the spatial sensitivity curve was highest at low spatial frequencies (0.05 and 0.11 cpd), and the sensitivity decreased steeply with an increase in the spatial frequency. RCs were not detectable at 3.46 cpd. The MTF decreased gradually with increases in the spatial frequencies and was 0.68 at 3.46 cpd. The reflectance pattern of the retinal intrinsic signal elicited by grating stimuli of different spatial frequencies was different from that of the MTF. This suggests that the intrinsic signal represents not only the response of the photoreceptors but also other neuronal or vascular changes in the retina.


Chemical, Biochemical, Cellular, and Physiological Characterization of Leucettinib-21, a Down Syndrome and Alzheimer's Disease Drug Candidate.
Leucettinibs are substituted 2-aminoimidazolin-4-ones (inspired by the marine sponge natural product Leucettamine B) developed as pharmacological inhibitors of DYRK1A (dual-specificity, tyrosine phosphorylation-regulated kinase 1A), a therapeutic target for indications such as Down syndrome and Alzheimer's disease. Leucettinib-21 was selected as a drug candidate following extensive structure/activity studies and multiparametric evaluations. We here report its physicochemical properties (X-ray powder diffraction, differential scanning calorimetry, stability, solubility, crystal structure) and drug-like profile. Leucettinib-21's selectivity (analyzed by radiometric, fluorescence, interaction, thermal shift, residence time assays) reveals DYRK1A as the first target but also some "off-targets" which may contribute to the drug's biological effects. Leucettinib-21 was cocrystallized with CLK1 and modeled in the DYRK1A structure. Leucettinib-21 inhibits DYRK1A in cells (demonstrated by direct catalytic activity and phosphorylation levels of Thr286-cyclin D1 or Thr212-Tau). Leucettinib-21 corrects memory disorders in the Down syndrome mouse model Ts65Dn and is now entering safety/tolerance phase 1 clinical trials.


Tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) and receptors in type 1, type 2 and type 17 inflammation in cross-sectional asthma study.
Tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) reportedly promotes, or conversely, resolves inflammation in asthma. In this study of TRAIL and cell receptors in sputum, bronchoalveolar lavage and biopsy from subjects in the Severe Asthma Research Program at Wake Forest, the high TRAIL group had significant increases in all leucocytes, and was associated with increased type 1, type 2 and type 17 cytokines, but not type 9 interleukin 9. Two variants at loci in the TRAIL gene were associated with higher sputum levels of TRAIL. Increased TRAIL decoy receptor R3/DcR1 was observed on sputum leucocytes compared with death receptor R1/DR4, suggesting reduced apoptosis and prolonged cellular inflammation.


Temperature robustness in Arabidopsis circadian clock models is facilitated by repressive interactions, autoregulation, and three-node feedbacks.
The biological interactions underpinning the Arabidopsis circadian clock have been systematically uncovered and explored by biological experiments and mathematical models. This is captured by a series of published ordinary differential equation (ODE) models, which describe plant clock dynamics in response to light/dark conditions. However, understanding the role of temperature in resetting the clock (entrainment) and the mechanisms by which circadian rhythms maintain a near-24 h period over a range of temperatures (temperature compensation) is still unclear. Understanding entrainment and temperature compensation may elucidate the principles governing the structure of the circadian clock network. Here we explore the design principles of the Arabidopsis clock and its responses to changes in temperature. We analyse published clock models of Arabidopsis, spanning a range of complexity, and incorporate temperature-dependent dynamics into the parameters of translation rates in these models, to discern which regulatory patterns may best explain clock function and temperature compensation. We additionally construct three minimal clock models and explore what key features govern their rhythmicity and temperature robustness via a series of random parameterisations. Results show that the highly repressive interactions between the components of the plant clock, together with autoregulation patterns and three-node feedback loops, are associated with circadian function of the clock in general, and enhance its robustness to temperature variation in particular. However, because the networks governing clock function vary with time due to light and temperature conditions, we emphasise the importance of studying plant clock functionality in its entirety rather than as a set of discrete regulation patterns.


Prescribing patterns and predictors of high-level polypharmacy in the elderly population: A prospective surveillance study from two teaching hospitals in India.
Polypharmacy has been reported to increase the risks for inappropriate prescribing, adverse drug reactions, geriatric syndromes, and morbidity and mortality in elderly populations in the United States and Europe. Data on prescribing patterns and polypharmacy in the elderly population in India are limited. The aims of this study were to assess prescribing patterns and to determine the predictors of high-level polypharmacy in the elderly population in 2 tertiary care hospitals in India. This prospective surveillance study used medical records from patients aged 60 to 95 years admitted to the medicine wards of the 2 tertiary care hospitals between January 2008 and June 2009. Data on medication prescribing from admission through discharge were reviewed. Diseases were coded using the World Health Organization (WHO) International Classification of Diseases, 10th Revision, and medications were coded using the WHO Anatomical, Therapeutic, and Chemical classification. Concordance of prescribing with the indications in the product labeling as listed in the American Hospital Formulary Services Drug Information 2007 was determined. The prevalences of polypharmacy (5-9 medications) and high-level polypharmacy (>or=10 medications) were determined. Bivariate analysis and multivariate logistic regression analysis were used to determine the influential predictors of high-level polypharmacy during hospital stays. Data from 814 patients were included (493 [60.6%] men, 321 [39.4%] women; median age, 66 years [range, 60-95 years]). Systemic antibacterials were the most commonly prescribed therapeutic class of medications (574 [70.5%]), and pantoprazole was the most commonly prescribed medication (498 [61.2%]). The majority (7/10 [70.0%]) of the most commonly prescribed medications were prescribed as indicated. Medications prescribed "off-label" included pantoprazole (432/498 [86.7%]), ceftriaxone (212/259 [81.9%]), and atorvastatin (109/237 [46.0%]). Polypharmacy and high-level polypharmacy were prescribed in 366 (45.0%) and 370 (45.5%) patients, respectively. On multivariate logistic regression analysis, multiple (>or=3) diagnoses (odds ratio [OR] = 1.55; 95% CI, 1.16-2.08; P = 0.003), angina pectoris (OR = 2.58; 95% CI, 1.50-4.37; P < 0.001), and a length of stay >or=10 days (10-<15 days, OR = 3.14; 95% CI, 2.09-4.71; P < 0.001; and >or=15 days, OR = 5.74; 95% CI, 2.43-13.51; P < 0.001) were found to be predictors of high-level polypharmacy during hospital stays. The campaign for rational drug use in hospitalized elderly patients in India should promote pantoprazole, ceftriaxone, and atorvastatin prescribing in concordance with their indications. Interventions to reduce the high-level polypharmacy in the elderly during their stays in tertiary care hospitals in India should focus on patients with >/=3 diagnoses, angina pectoris, and/or >or=10 days of hospital stay.


Leaf Dimorphism Of Microgramma Squamulosa (Polypodiaceae): a qualitative and quantitative analysis focusing on adaptations to epiphytism.
The epiphytic fern Microgramma squamulosa occurs in the Neotropics and shows dimorphic sterile and fertile leaves. The present study aimed to describe and compare qualitatively and quantitatively macroscopic and microscopic structural characteristics of the dimorphic leaves of M. squamulosa, to point more precisely those characteristics which may contribute to epiphytic adaptations. In June 2009, six isolated host trees covered by M squamulosa were selected close to the edge of a semi-deciduous seasonal forest fragment in the municipality of Novo Hamburgo, State of Rio Grande do Sul, Brazil. Macroscopic and microscopic analyzes were performed from 192 samples for each leaf type, and permanent and semi-permanent slides were prepared. Sections were observed under light microscopy using image capture software to produce illustrations and scales, as well as to perform quantitative analyses. Fertile and sterile leaves had no qualitative structural differences, being hypostomatous and presenting uniseriate epidermis, homogeneous chlorenchyma, amphicribal vascular bundle, and hypodermis. The presence of hypodermal tissue and the occurrence of stomata at the abaxial face are typical characteristics ofxeromorphic leaves. Sterile leaves showed significantly larger areas (14.80cm2), higher sclerophylly index (0.13g/cm2) and higher stomatal density (27.75stomata/mm2) than fertile leaves. The higher sclerophylly index and the higher stomatal density observed in sterile leaves are features that make these leaves more xeromorphic, enhancing their efficiency to deal with limited water availability in the epiphytic environment, compared to fertile leaves.


Oncogenic partnerships: EWS-FLI1 protein interactions initiate key pathways of Ewing's sarcoma.
Targeted therapy for cancer, which is specifically directed toward the cancer without any potential for effects outside of controlling the tumor, is a gold standard for treatment. Ewing's sarcoma contains the potential target EWS-FLI1, as a result of a pathognomonic chromosomal translocation. The EWS-FLI1 fusion protein includes the EWS domain, a potent transcriptional activator alongside the highly conserved FLI1 ets DNA-binding domain. Because of the combination of these domains, the EWS-FLI1 fusion protein acts as an aberrant transcription factor whose expression results in cellular transformation. EWS-FLI1 functions by binding to normal cellular protein partners in transcription and splicing, similar to how a virus would corrupt normal cellular machinery for virion production. Therefore, understanding the protein-protein interactions of EWS-FLI1 and the pathways that are regulated by these partnerships will inform both oncogenesis and therapeutics. This review describes the known protein partners and transcriptional targets of EWS-FLI1, while proposing strategies for exploiting these partnerships with targeted therapy.


Vascular endothelial growth factor regulates reendothelialization and neointima formation in a mouse model of arterial injury.
The rate of reendothelialization is critical in neointima formation after arterial injury. Vascular endothelial growth factor (VEGF), a potent endothelial mitogen, has been advocated for accelerating endothelial repair and preventing intimal hyperplasia after percutaneous coronary interventions. However, the precise mechanism of action of VEGF treatment and the physiologic role of endogenous VEGF after arterial injury are not well described. To better understand the role of VEGF in arterial repair, we overexpressed both VEGF and a soluble, chimeric VEGF receptor (VEGF-trap), which binds free VEGF with high affinity, in a mouse model of arterial injury. Four groups of C57BL/6 mice underwent denuding endothelial injury 1 day after systemic injection of recombinant adenovirus expressing (1) VEGF, (2) VEGF-trap, (3) VEGF plus VEGF-trap, or (4) control adenovirus. Circulating levels of adenovirus-encoded proteins were significantly elevated after gene transfer. VEGF overexpression accelerated reendothelialization and increased luminal endothelial cell proliferation 2 weeks after arterial injury (P<0.05), resulting in decreased neointima formation at 4 weeks compared with control (P<0.01). Cotreatment with VEGF-trap completely sequestered free VEGF and abrogated the beneficial effect of VEGF overexpression. Interestingly, sequestration of endogenous VEGF by VEGF-trap overexpression alone also led to delayed reendothelialization at 2 weeks (P<0.01) and increased neointima formation at 4 weeks (P<0.01). VEGF overexpression accelerated endothelial repair and inhibited neointima formation after arterial injury. Conversely, sequestration of exogenous and/or endogenous VEGF by VEGF-trap delayed reendothelialization and significantly increased neointima size. This demonstrates the therapeutic potential of VEGF but also emphasizes the important physiologic role of endogenous VEGF in vascular repair.


Ferrimonas senticii sp. nov., a novel gammaproteobacterium isolated from the mucus of a puffer fish caught in Kaneohe Bay, Hawai'i.
A novel species, strain P2S11T, was isolated from the mucus of a puffer fish caught off the coast of Kaneohe Bay, O'ahu, Hawai'i. A phylogenetic analysis based on 16S rRNA gene sequences showed that the novel strain was most closely related to Ferrimonas marina DSM 16917T and Ferrimonas balearica DSM 9799T with 93.5% and 82.9% sequence similarities, respectively, which established the novel strain as belonging to the genus Ferrimonas. The strain formed off-white coloured colonies on marine agar and cells were Gram-negative, non-motile rods. H2S was produced when strain P2S11T was grown on TSI medium with added salt. Strain P2S11T had a DNA G+C content of 54.9 mol% and the dominant fatty acids were C16:1omega9c, C16:0 and C17:1omega8c. On the basis of this polyphasic study, strain P2S11T (=ATCC BAA-1480T=DSM 18821T) represents a novel species of the genus Ferrimonas, for which the name Ferrimonas senticii sp. nov. is proposed.


A poly(amidoamine)-based polymeric nanoparticle platform for efficient in vivo delivery of mRNA.
The successful use of mRNA vaccines enabled and accelerated the development of several new vaccine candidates and therapeutics based on the delivery of mRNA. In this study, we developed bioreducible poly(amidoamine)-based polymeric nanoparticles (PAA PNPs) for the delivery of mRNA with improved transfection efficiency. The polymers were functionalized with chloroquinoline (Q) moieties for improved endosomal escape and further stabilization of the mRNA-polymer construct. Moreover, these PAAQ polymers were covalently assembled around a core of multi-armed ethylenediamine (Mw 800, 2 % w/w) to form a pre-organized polymeric scaffolded PAAQ (ps-PAAQ) as a precursor for the formation of the mRNA-loaded nanoparticles. Transfection of mammalian cell lines with EGFP mRNA loaded into these PNPs showed a favorable effect of the Q incorporation on GFP protein expression. Additionally, these ps-PAAQ NPs were co-formulated with PEG-polymer coatings to shield the positive surface charge for increased stability and better in vivo applicability. The ps-PAAQ NPs coated with PEG-polymer displayed smaller particle size, electroneutral surface charge, and higher thermal stability. Importantly, these nanoparticles with both Q and PEG-polymer coating induced significantly higher luciferase activity in mice muscle than uncoated ps-PAAQ NPs, following intramuscular injection of PNPs loaded with luciferase mRNA. The developed technology is broadly applicable and holds promise for the development of new nucleotide-based vaccines and therapeutics in a range of infectious and chronic diseases.


Implementing economic evaluation in simulation-based medical education: challenges and opportunities.
Simulation-based medical education (SBME) is now ubiquitous at all levels of medical training. Given the substantial resources needed for SBME, economic evaluation of simulation-based programmes or curricula is required to demonstrate whether improvement in trainee performance (knowledge, skills and attitudes) and health outcomes justifies the cost of investment. Current literature evaluating SBME fails to provide consistent and interpretable information on the relative costs and benefits of alternatives. Economic evaluation is widely applied in health care, but is relatively scarce in medical education. Therefore, in this paper, using a focus on SBME, we define economic evaluation, describe the key components, and discuss the challenges associated with conducting an economic evaluation of medical education interventions. As a way forward to the rigorous and state of the art application of economic evaluation in medical education, we outline the steps to gather the necessary information to conduct an economic evaluation of simulation-based education programmes and curricula, and describe the main approaches to conducting an economic evaluation. A properly conducted economic evaluation can help stakeholders (i.e., programme directors, policy makers and curriculum designers) to determine the optimal use of resources in selecting the modality or method of assessment in simulation. It also helps inform broader decision making about allocation of scarce resources within an educational programme, as well as between education and clinical care. Economic evaluation in medical education research is still in its infancy, and there is significant potential for state-of-the-art application of these methods in this area.


Towards the Oral Treatment of Ileo-Colonic Inflammatory Bowel Disease with Infliximab Tablets: Development and Validation of the Production Process.
Infliximab (IFX) is an intravenously administered monoclonal antibody antagonizing the effects of tumor necrosis factor-alpha (TNF) systemically and is efficacious in the treatment of inflammatory bowel disease (IBD). However, studies suggest that the anti-inflammatory effects result from local immunomodulation in the inflamed regions. Furthermore, topical inhibition of TNF in IBD ameliorates inflammation. We therefore hypothesized that orally administered IFX targeted to the ileo-colonic region in IBD may be an efficacious new treatment option. This study describes the development and validation of the production process of ileo-colonic-targeted 5 mg IFX tablets (ColoPulse-IFX) intended for the oral treatment of IBD by means of producing three consecutive validation batches (VAL1, VAL2, and VAL3, respectively). UV-VIS spectroscopy, HPLC-SEC analysis (content, fragments, aggregates), fluorescence spectroscopy (tertiary protein structure), and ELISA (potency) showed no noticeable deviations of IFX compounded to ColoPulse-IFX compared to fresh IFX stock. The average ± SD (n = 10) IFX content of VAL1, VAL2, and VAL3 was 96 ± 2%, 97 ± 3%, and 96 ± 2%, respectively, and complied with the European Pharmacopeia (Ph. Eur.) requirements for Content Uniformity. The average ± SD (n = 3) ColoPulse-IFX potency was 105 ± 4%, 96 ± 4%, and 97 ± 5%, respectively, compared to fresh IFX stock. The IFX release profile from the tablet core was complete (≥85%) after 10 min in simulated ileum medium. The in vitro coating performance of ColoPulse-IFX showed that the formulation was targeted to the simulated ileo-colonic region. Stability data showed that ColoPulse-IFX was stable for up to 6 months stored at 25 °C/60% RH. Based on these results, the production process can be considered validated and its application is discussed in light of the rationale and available evidence for the topical treatment of IBD with IFX.


Increased oxidative stress in preschool children exposed to passive smoking.
To study the effect of passive cigarette smoking on plasma oxidative and antioxidative status in passive smoking preschool children and to compare them with controls. Thirty-four passive smoking (five to 50 cigarettes per day) preschool children (study group) and 32 controls who had never been exposed to cigarette smoke were randomly chosen from children aged from 4 to 6 years. Urinary cotinine and plasma indicators of oxidative and antioxidative status, i.e., total oxidant status (TOS), total antioxidant capacity (TAC), and oxidative stress index (OSI), were determined. Mean environmental cigarette consumption was 22±13 cigarettes per day in passive smoking children. Mean urinary cotinine levels were 77.6±41.4 ng/mL and 11.9±2.3 ng/mL in the study and control groups, respectively (p < 0.001). Mean plasma TAC levels were 0.95±0.13 mmol Trolox equivalent/L and 1.01±0.09 mmol Trolox equivalent/L, respectively (p = 0.039). Mean plasma TOS levels were 28.6±7.9 µmol H2O2 equivalent/L and 18.5±6.3 µmol H2O2 equivalent/L, respectively (p < 0.001). Mean OSI levels were 3.08±0.98 arbitrary units and 1.84±0.64 arbitrary units, respectively (p < 0.001). A small amount of cigarette smoke (five to 10 cigarettes per day) causes considerable oxidative stress. There were significant correlations between number of cigarettes consumed and oxidant status and OSI levels. Passive smoke is a potent oxidant in preschool children. Its deleterious effects are not limited just to heavy passive smoking, but also occur with exposure to small amounts of smoke.


Lost alpha Faraday cup foil noise characterization during Joint European Torus plasma post-processing analysis.
Capacitive plasma pickup is a well-known and difficult problem for plasma-facing edge diagnostics. This problem must be addressed to ensure an accurate and robust interpretation of the real signal measurements vs noise. The Faraday cup fast ion loss detector array of the Joint European Torus (JET) is particularly prone to this issue and can be used as a testbed to prototype solutions. The issue of separation and distinction between warranted fast ion signal and electromagnetic plasma noise has traditionally been solved with hardware modifications, but a more versatile post-processing approach is of great interest. This work presents post-processing techniques to characterize the signal noise. While hardware changes and advancements may be limited, the combination with post-processing procedures allows for more rapid and robust analysis of measurements. The characterization of plasma pickup noise is examined for alpha losses in a discharge from JET's tritium campaign. In addition to highlighting the post-processing methodology, the spatial sensitivity of the detector array is also examined, which presents significant advantages for the physical interpretation of fast ion losses.


Role of calcitonin gene-related peptide-containing nerves in the vascular adrenergic neurotransmission.
The role of calcitonin gene-related peptide (CGRP)-containing nerves in adrenergic vasoconstrictor response to periarterial nerve stimulation (PNS) was investigated in mesenteric vascular beds isolated from rats, which were perfused with Krebs' solution (5 ml/min). In perfused mesenteric vascular beds, PNS (1-12 Hz, for 30 sec) caused a frequency-dependent increase in the perfusion pressure, which was prevented by 50 nM prazosin and abolished by 300 nM tetrodotoxin and 5 microM guanethidine. Bolus infusion of norepinephrine (NE, 0.5 and 1 nmol) also produced a pressor response which was blocked by 50 nM prazosin. In the preparation treated with 500 nM capsaicin for 20 min, pressor responses to PNS of 1 to 8 Hz were potentiated significantly. The pressor responses to NE infusion also were enhanced in both the preparations treated with and without capsaicin. In the capsaicin-untreated preparation contracted by 7 mM methoxamine in the presence of 5 microM guanethidine, PNS (1-12 Hz) caused a frequency-dependent vasodilation, which was abolished by 300 nM tetrodotoxin. However, no vasodilator response to PNS was observed in the preparation treated with capsaicin. In the contracted preparation, bolus infusion of rat CGRP (10 and 100 pmol) produced a marked long-lasting vasodilation which mimicked the PNS-evoked vasodilation, whereas neither bolus infusion of substance P (1 and 10 nmol), neurokinin A (1 and 10 nmol) nor neurokinin B (1 and 10 nmol) produced relaxation. In the preparation labeled with [3H]NE, the PNS (4 Hz)-evoked 3H release was not altered after capsaicin treatment, whereas the pressor response to PNS was potentiated significantly.(ABSTRACT TRUNCATED AT 250 WORDS)


Do chronic urinary tract infections induce cancer in the rat fed nitrate and aminopyrine?
Seven out of one hundred male rats in the experimental group developed malignant tumours after 18 months, during which period they were given nitrate and aminopyrine in the drinking water and had chronic urinary tract infections induced by known strains of E. coli. In the control animals, no lesions were observed in the two un-infected groups given nitrosamine precursors in the drinking water or water alone. However, two out of fifty control animals in the infected group, not administered nitrosamine precursors, developed malignant lymphoma. All rats were maintained on Purina rat chow. Our data appear to suggest that chronic urinary tract infection did induce tumour production in the rat.


Pre-warming before general anesthesia with isoflurane delays the onset of hypothermia in rats.
General anesthesia causes hypothermia by impairing normal thermoregulatory mechanisms. When using inhalational anesthetic agents, Redistribution of warm blood from the core to the periphery is the primary mechanism in the development of hypothermia and begins following induction of anesthesia. Raising skin temperature before anesthesia reduces the temperature gradient between core and periphery, decreasing the transfer of heat. This prospective, crossover study (n = 17 adult male and female SD rats) compared three treatment groups: PW1% (pre-warming to increase core temperature 1% over baseline), PW40 (pre-warming to increase core temperature to 40°C) and NW (no warming). The PW1% group was completed first to ensure tolerance of pre-warming. Treatment order was then randomized and alternated after a washout period. Once target temperature was achieved, anesthesia was induced and maintained with isoflurane in oxygen without further external temperature support. Pre-warming was effective at delaying the onset of hypothermia, with a significant difference between PW1% (12.4 minutes) and PW40 (19.3 minutes, p = 0.0044 (95%CI -12 to -2.2), PW40 and NW (7.1 minutes, p < 0.0001 (95%CI 8.1 to 16.0) and PW1% and NW (p = 0.003, 95%CI 1.8 to 8.7). The rate of heat loss in the pre-warmed groups exceed that of the NW group: PW1% versus NW (p = 0.005, 95%CI 0.004 to 0.027), PW40 versus NW (p < 0.0001, 95%CI 0.014 to 0.036) and PW1% versus PW40 (p = 0.07, 95%CI -0.021 to 0.00066). Pre-warming alone confers a protective effect against hypothermia during volatile anesthesia; however, longer duration procedures would require additional heating support.


Biomedical Informatics Workforce in Croatia: Qualitative Analysis of Teachers' Opinions on Needs and Employment Opportunities.
We report a qualitative analysis of higher education biomedical informatics (BMI) teachers' opinions on the needs for BMI workforce and employment opportunities in the Croatian health care system. Needs were perceived as considerable; however this was not coupled with adequate employment opportunities. Barriers were identified to correcting this imbalance, including the failure of the health care system planners to recognise BMI experts as an important part of the workforce and lack of adequate educational opportunities.


Towards enhanced and interpretable clustering/classification in integrative genomics.
High-throughput technologies have led to large collections of different types of biological data that provide unprecedented opportunities to unravel molecular heterogeneity of biological processes. Nevertheless, how to jointly explore data from multiple sources into a holistic, biologically meaningful interpretation remains challenging. In this work, we propose a scalable and tuning-free preprocessing framework, Heterogeneity Rescaling Pursuit (Hetero-RP), which weighs important features more highly than less important ones in accord with implicitly existing auxiliary knowledge. Finally, we demonstrate effectiveness of Hetero-RP in diverse clustering and classification applications. More importantly, Hetero-RP offers an interpretation of feature importance, shedding light on the driving forces of the underlying biology. In metagenomic contig binning, Hetero-RP automatically weighs abundance and composition profiles according to the varying number of samples, resulting in markedly improved performance of contig binning. In RNA-binding protein (RBP) binding site prediction, Hetero-RP not only improves the prediction performance measured by the area under the receiver operating characteristic curves (AUC), but also uncovers the evidence supported by independent studies, including the distribution of the binding sites of IGF2BP and PUM2, the binding competition between hnRNPC and U2AF2, and the intron-exon boundary of U2AF2 [availability: https://github.com/younglululu/Hetero-RP].


Problems of tic douloureux (trigeminal neuralgia) management.
A review of experience with the management of tic douloureux (trigeminal neuralgia) at Groote Schuur Hospital from 1948 to 1972 shows that thermocoagulation, fractional section of the Gasserian ganglion and phenol injection were the methods most frequently used. After 1972, carbamazepine was found to be an effective treatment but follow-up was difficult. The alternative surgical treatments of posterior fossa exploration of the trigeminal nerve for release of vascular compression and radiofrequency coagulation of the Gasserian ganglion are discussed. Since 1979 radiofrequency coagulation of the Gasserian ganglion has been employed when carbamazepine has failed. The technique and results are described.


Neuropilin-1 in transplantation tolerance.
In the immune system, Neuropilin-1 (Nrp1) is a molecule that plays an important role in establishing the immunological synapse between dendritic cells (DCs) and T cells. Recently, Nrp1 has been identified as a marker that seems to distinguish natural T regulatory (nTreg) cells, generated in the thymus, from inducible T regulatory (iTreg) cells raised in the periphery. Given the crucial role of both nTreg and iTreg cells in the generation and maintenance of immune tolerance, the ability to phenotypically identify each of these cell populations in vivo is needed to elucidate their biological properties. In turn, these properties have the potential to be developed for therapeutic use to promote immune tolerance. Here we describe the nature and functions of Nrp1, including its potential use as a therapeutic target in transplantation tolerance.


An experimental and computational study of the effect of aqueous solution on the multiphoton ionisation photoelectron spectrum of phenol.
We revisit the photoelectron spectroscopy of aqueous phenol in an effort to improve our understanding of the impact of inhomogeneous broadening and inelastic scattering on solution-phase photoelectron spectra. Following resonance-enhanced multiphoton ionisation via the 11ππ* and 11πσ* states of phenol, we observe 11ππ*-D0/D1 ionisation and competing direct S0-D0/D1 ionisation. Following resonance-enhanced multiphoton ionisation via the 21ππ* state, we observe the signature of solvated electrons. By comparing the photoelectron spectra of aqueous phenol with those of gas-phase phenol, we find that inelastic scattering results in peak shifts with similar values to those that have been observed in photoelectron spectra of solvated electrons, highlighting the need for a robust way of deconvoluting the effect of inelastic scattering from liquid-phase photoelectron spectra. We also present a computational strategy for calculating vertical ionisation energies using a quantum-mechanics/effective fragmentation potential (QM/EFP) approach, in which we find that optimising the configurations obtained from molecular dynamics simulations and using the [phenol·(H2O)5]QM[(H2O)n≥250]EFP (B3LYP/aug-cc-pvdz) method gives good agreement with experiment.


Simultaneous ultra-high-pressure liquid chromatography-tandem mass spectrometry determination of amphetamine and amphetamine-like stimulants, cocaine and its metabolites, and a cannabis metabolite in surface water and urban wastewater.
An ultra-high-pressure liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method has been developed for the simultaneous quantification and confirmation of 11 basic/acidic illicit drugs and relevant metabolites in surface and urban wastewater at ng/L levels. The sample pre-treatment consisted of a solid-phase extraction using Oasis MCX cartridges. Analyte deuterated compounds were used as surrogate internal standards (except for norbenzoylecgonine and norcocaine) to compensate for possible errors resulting from matrix effects and those associated to the sample preparation procedure. After SPE enrichment, the selected drugs were separated within 6min under UHPLC optimized conditions. To efficiently combine UHPLC with MS/MS, a fast-acquisition triple quadrupole mass analyzer (TQD from Waters) in positive-ion mode (ESI+) was used. The excellent selectivity and sensitivity of the TQD analyzer in selected reaction monitoring mode allowed quantification and reliable identification at the LOQ levels. Satisfactory recoveries (70-120%) and precision (RSD<20%) were obtained for most compounds in different types of water samples, spiked at two concentration levels [limit of quantification (LOQ) and 10LOQ]. Thus, surface water was spiked at 30 ng/L and 300 ng/L (amphetamine and amphetamine-like stimulants), 10 ng/L and 100 ng/L (cocaine and its metabolites), 300 ng/L and 3000 ng/L (tetrahydrocannabinol-COOH). Recovery experiments in effluent and influent wastewater were performed at spiking levels of three and fifteen times higher than the levels spiked in surface water, respectively. The validated method was applied to urban wastewater samples (influent and effluent). The acquisition of three selected reaction monitoring transitions per analyte allowed positive findings to be confirmed by accomplishment of ion ratios between the quantification transition and two additional specific confirmation transitions. In general, drug consumption increased in the weekends and during an important musical event. The highest concentration levels were 27.5 microg/L and 10.5 microg/L, which corresponded to 3,4-methylenedioxymethamphetamine (MDMA, or ecstasy) and to benzoylecgonine (a cocaine metabolite), respectively. The wastewater treatment plants showed good removal efficiency (>99%) for low levels of illicit drugs in water, but some difficulties were observed when high drug levels were present in wastewaters.


Bis(2,2'-bipyridyl-κ(2) N,N')(sulfato-κ(2) O,O')nickel(II) 2.5-hydrate.
The title compound, [Ni(SO4)(C10H8N2)2]·2.5H2O, is a nickel(II) complex with a distorted octa-hedral coordination geometry. The Ni(II) atom is bonded by two O atoms of the bidentate chelating sulfate ligand and the four N atoms of two chelating 2,2'-bi-pyridine ligands. The Ni-N bond lengths range from 2.059 (3) to 2.075 (3) Å and the Ni-O bond lengths are 2.098 (3) and 2.123 (3) Å. The bipyridyl ligands are both close to planar (r.m.s. deviations of 0.254 and 0.0572 Å) and are almost orthogonal, making a dihedral angle of 82.77 (1)°. In the crystal, the complex and water mol-ecules are connected by O-H⋯O hydrogen bonds. Inter-estingly, six water mol-ecules form a chain linking two complex mol-ecules via sulfate O atoms. There are also stacking inter-actions between the aromatic rings of neighbouring 2,2'-bi-pyridine ligands with shortest non-covalent contacts of 3.268 (6), 3.393 (6) and 3.435 (5) Å. One of the three unique water molecules shows half-occupation.


Effect of variations in conditions of incubation upon inhibition of Staphylococcus aureus by Pediococcus cerevisiae and Streptococcus lactis.
The effects of pH, temperature, proportion of Staphylococcus aureus in the inoculum, various strains of effector organism, and various strains of S. aureus were examined for their influence on interactions between staphylococci and effector organisms in associative culture. In general, small changes in pH had little effect upon either growth of S. aureus or production of enterotoxin in associative culture. Inhibition of growth of S. aureus caused by effector organisms was much greater at 25 than at 30 C. Proportion of S. aureus in the inoculum greatly affected both growth of the staphylococci and production of enterotoxin. Only slight differences were found between strains of either effector organism or S. aureus which affected the interactions in associative culture.


Unbiased computation of transition times by pathway recombination.
In many systems, the time scales of the microscopic dynamics and macroscopic dynamics of interest are separated by many orders of magnitude. Examples abound, for instance, nucleation, protein folding, and chemical reactions. For these systems, direct simulation of phase space trajectories does not efficiently determine most physical quantities of interest. The past decade has seen the advent of methods circumventing brute force simulation. For most dynamical quantities, these methods all share the drawback of systematical errors. We present a novel method for generating ensembles of phase space trajectories. By sampling small pieces of these trajectories in different phase space domains and piecing them together in a smart way using equilibrium properties, we obtain physical quantities such as transition times. This method does not have any systematical error and is very efficient; the computational effort to calculate the first passage time across a free energy barrier does not increase with the height of the barrier. The strength of the method is shown in the Ising model. Accurate measurements of nucleation times span almost ten orders of magnitude and reveal corrections to classical nucleation theory.


[Level of evidence for therapeutic drug monitoring of MPA in hematopoietic stem cell transplantation].
Mycophenolic acid (MPA) is more and more used to prevent GVHD (Graft Versus Host Disease) during hematopoietic stem cell transplantation with reduce-intensity conditioning. If several facts argue in favor of therapeutic drug monitoring, the used pharmacokinetic parameter is to be defined. Especially, the choice between total or ultrafilterable MPA is still under debate even if therapeutic drug monitoring seems to be more practicable with total MPA. The role of other factors implied in GVHD occurrence are also to be assessed in studies which aim at assessing therapeutic drug monitoring of MPA in such situation. For theses reasons, the level evidence of MPA as GVHD prophylaxis during hematopoietic stem cell transplantation with reduce-intensity conditioning is potentially useful.


Synthesis, spectroscopic and electrochemical characterization, and DFT study of seventeen C70(CF3)n derivatives (n=2, 4, 6, 8, 10, 12).
Eight new C70(CF3)n derivatives (n=2, 6, 10, 12) have been synthesized and characterized by UV/Vis and 19F NMR spectroscopy, cyclic voltammetry, and quantum chemical calculations at the DFT level of theory. Nine previously known derivatives of C70(CF3)n with n=2-12 were also studied by cyclic voltammetry (and seven of them by UV/Vis spectroscopy for the first time). Most of the 17 compounds exhibited two or three reversible reductions at scan rates from 20 mV s(-1) up to 5.0 V s(-1). In general, reduction potentials for the 0/- couple are shifted anodically relative to the C70 0/-) couple. However, the 0/- E1/2 values for a given composition are strongly dependent on the addition pattern of the CF3 groups. The data show that the addition pattern is as important, if not more important in some cases, than the number of substituents, n, in determining E1/2 values. An analysis of the DFT-predicted LUMOs indicates that addition patterns that have non-terminal double bonds in pentagons result in derivatives that are strong electron acceptors.


The structure and activation of substrate water molecules in Sr(2+)-substituted photosystem II.
The mechanism of solar water oxidation by photosystem II (PSII) is of fundamental interest and it is the object of extensive studies both in the past and present. The solar water oxidation reaction of PSII occurs in the oxygen-evolving complex (OEC). The OEC consists of a tetranuclear manganese calcium-oxo (Mn4Ca-oxo) cluster that is surrounded by amino acid residues and inorganic cofactors. The role of the Ca(2+) ion in the water oxidation reaction is one of the most interesting questions that is yet to be answered. In this study, we probe the structural and functional differences induced by metal ion substitution in the Mn4Ca-oxo cluster by substituting the Ca(2+) ion in the OEC by a Sr(2+) ion. We apply two-dimensional (2D) hyperfine sublevel correlation (HYSCORE) spectroscopy to detect weak magnetic interactions between the paramagnetic Mn4Sr-oxo cluster and the surrounding protons in the S2 state of the OEC of Sr(2+)-substituted PSII. We identify three groups of protons that are magnetically interacting with the Mn4Sr-oxo cluster. Using the recently reported 1.9 Å resolution X-ray structure of the OEC in the S1 state [Umena et al.] and the high-resolution 2D HYSCORE spectroscopy studies of the S2 state of the OEC of Ca(2+)-containing PSII [Milikisiyants et al., Energy Environ. Sci., 2012, 5, 7747], we discuss the assignments of the three groups of protons that are magnetically coupled to the Mn4Sr-oxo cluster. Since hyperfine interactions are highly sensitive to small perturbations in the electronic and geometric structure of paramagnetic centers, a comparison of the 2D HYSCORE spectra of Sr(2+)-substituted and Ca(2+)-containing PSII allows us to draw important conclusions with respect to the structure of the substrate water molecules in the OEC and the role of the Ca(2+) ion in the water oxidation reaction. In addition, for the first time, we determine the experimental value of the spin projection factor for the Mn(III) ion of the Mn4Ca-oxo cluster as ρ1 = ±1.7 from the assignment of the hyperfine interaction of the paramagnetic cluster with the protons of the D1-His332 residue of PSII.


Surface relocation of alpha 6 beta 4 integrins and assembly of hemidesmosomes in an in vitro model of wound healing.
A transmembrane extracellular matrix receptor of the integrin family, alpha 6 beta 4, is a component of the hemidesmosome, an adhesion complex of importance in epithelial cell-connective tissue attachment (Stepp, M. A., S. Spurr-Michaud, A. Tisdale, J. Elwell, and I. K. Gipson. 1990. Proc. Natl. Acad. Sci. USA. 87:8970-8974; Jones, J. C. R., M. A. Kurpakus, H. M. Cooper, and V. Quaranta. 1991. Cell Regulation. 2:427-438). Cytosolic components of hemidesmosomes include bullous pemphigoid (BP) antigens while extracellular components include a 125-kD component of anchoring filaments (CAF) and collagen type VII-containing anchoring fibrils. We have monitored the incorporation of the alpha 6 beta 4 integrins into forming hemidesmosomes in an in vitro wound-healing explant model. In epithelial cells recently migrated from the edges of unwounded sites over bare connective tissue, alpha 6 beta 4 first appears along the entire cell surface. At this stage, these cells contain little or no cytosolic hemidesmosomal components, at least as detectable by immunofluorescence using BP autoantibodies, whereas they are already positive for laminin and CAF. At a later stage, as cells become positive for cytosolic hemidesmosome components such as BP antigens as well as collagen type VII, alpha 6 beta 4 becomes concentrated along the basal pole of the epithelial cell where it abuts the connective tissue of the explant. Polyclonal antibodies to beta 4 do not interfere with the migration of epithelial cells in the explant. However, they prevent assembly of hemidesmosomal complexes and inhibit expression of collagen type VII in cells that have migrated over wound areas. In addition, they induce disruption of established hemidesmosomes in nonmigrating cells of the unwounded area of the explant. Monoclonal antibodies to alpha 6 have a more dramatic effect, since they completely detach epithelial cells in the unwounded area of the explant. Antibodies to CAF also detach epithelial cells in unwounded areas, apparently by inducing separation between epithelium and connective tissue at the lamina lucida of the basement membrane zone. These results suggest a model whereby polarization of alpha 6 beta 4 to the basal surface of the cells, perhaps induced by a putative anchoring filament-associated ligand, triggers assembly of hemidesmosome plaques.


Sulfasalazine for rheumatoid arthritis.
To estimate the short-term efficacy and toxicity of sulfasalazine for the treatment of rheumatoid arthritis (RA). We searched the Cochrane Musculoskeletal Group trials register, and Medline, up to July 1997, using the search strategy developed by the Cochrane Collaboration (Dickersin 1994). The search was complemented with bibliography searching of the reference list of the trials retrieved from the electronic search. Key experts in the area were contacted for further published and unpublished articles. All randomized controlled trials (RCTs) and controlled clinical trials (CCTs) comparing sulfasalazine against placebo in patients with RA. Two reviewers determined the studies to be included based on inclusion and exclusion criteria (GW, MSA). Data were independently abstracted by two reviewers (EB, MSA), and checked by a third reviewer (BS) using a pre-developed form for the rheumatoid arthritis sub-group of the Cochrane Musculoskeletal Group. The same two reviewers, using a validated scale (Jadad 1996) assessed the methodological quality of the RCTs and CCTs independently. Rheumatoid arthritis outcome measures were extracted from the publications. The pooled analysis was performed using standardized mean differences (SMDs) for joint counts, pain, and global and functional assessments. Weighted mean differences (WMDs) were used for erythrocyte sedimentation rate (ESR). Toxicity was evaluated with pooled odds ratios (OR) for withdrawals. A chi-square test was used to assess heterogeneity among trials. Fixed effects models were used throughout and random effects for outcomes showing heterogeneity. Six trials, including 468 patients were included. A statistically significant benefit was observed for sulfasalazine when compared to placebo for tender and swollen joint scores, pain and ESR. The standardized weighted mean difference between treatment and placebo was -0.49 for tender and swollen joint scores, and -0.42 for pain. The difference for ESR was -17.6mm. Withdrawals from adverse reactions were significantly higher in the sulfasalazine group (OR=3.0). Patients receiving placebo were four times more likely to discontinue treatment because of lack of efficacy than patients receiving sulfasalazine. Sulfasalazine appears to have a clinically and statistically significant benefit on the disease activity of patients with RA. Its effects on overall health status and radiological progression are not clear at this time, but would appear to be modest.


[Fat-embolic vascular occlusions in avian lungs. Research of histologically determinable neutral fats in parrotlike birds (Psittaciformes)].
In 50 flying birds (49 parrot-like, one peacock), the content of fat-embolic vascular occlusions was determined. In 22 cases, fat-embolic occlusions of low intensity were found in pulmonary capillaries. There were no occlusions in renal vessels. The fat embolies had no influence on a lethal outcome. There was no statistically significant correlation between the fatty infiltration of the liver, the fat content of the bone marrow, and the frequency of fat-embolic occlusion of blood vessels. In 18 cases, mycoses were the cause of death. These infections did not influence the number of fat-embolic vascular occlusions.