Fibrous wound dressings encapsulating essential oils as natural antimicrobial agents.
Preventing infections is one of the main focuses of wound care. The colonisation of wounds by microorganisms can in fact have negative consequences on the healing process, delaying it. Here, we propose the use of essential oils as natural antimicrobial agents for cellulose-based fibrous dressings. We demonstrate the production of composite electrospun fibres that effectively encapsulate three different types of essential oils (cinnamon, lemongrass and peppermint). The fibrous scaffolds are able to inhibit the growth of Escherichia coli, even when small amounts of essential oils were used. At the same time, they are not cytotoxic, as proved by biocompatibility assays on skin cell models. The created dressings are promising as advanced biomedical devices for topical treatments.


Daytime melatonin administration in elderly good and poor sleepers: effects on core body temperature and sleep latency.
Melatonin has been shown to have hypnotic and hypothermic effects in young adults and has been proposed as treatment for insomnia. However, the hypnotic and thermoregulatory effects of melatonin remain to be simultaneously investigated for aged good and poor sleepers. The aim of this study was to explore the short-term effects of exogenous oral daytime melatonin on core body temperature, sleep latency, and subjective vigor and affect in aged women. Twelve sleep maintenance insomniacs and 10 good sleeping postmenopausal female subjects [mean (SD) age = 65.2 (7.4) years] participated in a double-blind, crossover study in which they received a capsule containing either melatonin (5 mg) or a placebo at 1400 hours. Continuous core body temperature and hourly multiple sleep latency tests (MSLT) were collected from 1100-2030 hours. Self-reported estimates of global vigor (sleepiness) and affect were collected prior to each MSLT using visual analog scales. Comparison of good and poor sleepers failed to reveal any significant differences in core body temperature, sleep latency, or subjective vigor and affect. However, for both groups combined, melatonin administration [absolute postadministration mean (SEM) = 36.9 (0.05) degrees C] significantly lowered core body temperature compared with placebo [37.1 (0.05) degrees C]. Similarly, melatonin administration significantly reduced latency to stage 1 (SOL1) and stage 2 (SOL2) [absolute postadministration mean SOL1 = 20.1 (1.7) and SOL2 = 20.7 (1.6) minutes] compared with placebo [SOL1 = 24.3 (1.2) and SOL2 = 25.2 (1.1) minutes]. Treatment had no significant effect on either vigor or affect. Overall, our results suggest that although short-term exogenous oral daytime melatonin has significant hypothermic and hypnotic effects in aged women, the size of the effects is modest.


Neodymium-140 DOTA-LM3: Evaluation of an In Vivo Generator for PET with a Non-Internalizing Vector.
140Nd (t1/2 = 3.4 days), owing to its short-lived positron emitting daughter 140Pr (t1/2 = 3.4 min), has promise as an in vivo generator for positron emission tomography (PET). However, the electron capture decay of 140Nd is chemically disruptive to macrocycle-based radiolabeling, meaning that an in vivo redistribution of the daughter 140Pr is expected before positron emission. The purpose of this study was to determine how the delayed positron from the de-labeled 140Pr affects preclinical imaging with 140Nd. To explore the effect, 140Nd was produced at CERN-ISOLDE, reacted with the somatostatin analogue, DOTA-LM3 (1,4,7,10- tetraazacyclododecane, 1,4,7- tri acetic acid, 10- acetamide N - p-Cl-Phecyclo(d-Cys-Tyr-d-4-amino-Phe(carbamoyl)-Lys-Thr-Cys)d-Tyr-NH2) and injected into H727 xenograft bearing mice. Comparative pre- and post-mortem PET imaging at 16 h postinjection was used to quantify the in vivo redistribution of 140Pr following 140Nd decay. The somatostatin receptor-positive pancreas exhibited the highest tissue accumulation of 140Nd-DOTA-LM3 (13% ID/g at 16 h) coupled with the largest observed redistribution rate, where 56 ± 7% (n = 4, mean ± SD) of the in situ produced 140Pr washed out of the pancreas before decay. Contrastingly, the liver, spleen, and lungs acted as strong sink organs for free 140Pr3+. Based upon these results, we conclude that 140Nd imaging with a non-internalizing vector convolutes the biodistribution of the tracer with the accumulation pattern of free 140Pr. This redistribution phenomenon may show promise as a probe of the cellular interaction with the vector, such as in determining tissue dependent internalization behavior.


Cancer management from a chronic gastrointestinal function perspective.
Bowel dysfunction in cancer is a significant and challenging issue for both clinicians and patients. As cancer survival improves, the impact of gastrointestinal symptoms on quality of life is of ever-increasing relevance. This review aims to provide an overview of the common gastrointestinal complaints seen in cancer sufferers and discuss the principles of management and up to date treatment options available.


Contrasting the optical properties of the different isomers of oligophenylene.
We use a combination of Time-Dependent Density Functional Theory (TD-DFT) and approximate Coupled Cluster Theory (RI-CC2) to compare trends in the optical gap and fluorescence energies of ortho-, meta- and para-oligomers of phenylene. We find that RI-CC2 and TD-DFT calculations using three different commonly employed XC-potentials (B3LYP, BHLYP and CAM-B3LYP) generally give consistent predictions. Most importantly, the fluorescence energy of m-phenylene is predicted to be independent of oligomer length, the fluorescence energy of p-phenylene to decrease with oligomer length and that of o-phenylene to increase. The origins of these differences in behaviour between the different isomers are analysed and found to stem from a subtle combination of steric and electronic factors.


Crystal structure of the eIF4A-PDCD4 complex.
Tumor suppressor programmed cell death protein 4 (PDCD4) inhibits the translation initiation factor eIF4A, an RNA helicase that catalyzes the unwinding of secondary structure at the 5'-untranslated region of mRNAs and controls the initiation of translation. Here, we determined the crystal structure of the human eIF4A and PDCD4 complex. The structure reveals that one molecule of PDCD4 binds to the two eIF4A molecules through the two different binding modes. While the two MA3 domains of PDCD4 bind to one eIF4A molecule, the C-terminal MA3 domain alone of the same PDCD4 also interacts with another eIF4A molecule. The eIF4A-PDCD4 complex structure suggests that the MA3 domain(s) of PDCD4 binds perpendicular to the interface of the two domains of eIF4A, preventing the domain closure of eIF4A and blocking the binding of RNA to eIF4A, both of which are required events in the function of eIF4A helicase. The structure, together with biochemical analyses, reveals insights into the inhibition mechanism of eIF4A by PDCD4 and provides a framework for designing chemicals that target eIF4A.


Intracellular trafficking of silicon particles and logic-embedded vectors.
Mesoporous silicon particles show great promise for use in drug delivery and imaging applications as carriers for second-stage nanoparticles and higher order particles or therapeutics. Modulation of particle geometry, surface chemistry, and porosity allows silicon particles to be optimized for specific applications such as vascular targeting and avoidance of biological barriers commonly found between the site of drug injection and the final destination. In this study, the intracellular trafficking of unloaded carrier silicon particles and carrier particles loaded with secondary iron oxide nanoparticles was investigated. Following cellular uptake, membrane-encapsulated silicon particles migrated to the perinuclear region of the cell by a microtubule-driven mechanism. Surface charge, shape (spherical and hemispherical) and size (1.6 and 3.2 microm) of the particle did not alter the rate of migration. Maturation of the phagosome was associated with an increase in acidity and acquisition of markers of late endosomes and lysosomes. Cellular uptake of iron oxide nanoparticle-loaded silicon particles resulted in sorting of the particles and trafficking to unique destinations. The silicon carriers remained localized in phagosomes, while the second stage iron oxide nanoparticles were sorted into multi-vesicular bodies that dissociated from the phagosome into novel membrane-bound compartments. Release of iron from the cells may represent exocytosis of iron oxide nanoparticle-loaded vesicles. These results reinforce the concept of multi-functional nanocarriers, in which different particles are able to perform specific tasks, in order to deliver single- or multi-component payloads to specific sub-cellular compartments.


Comparative study of methods for WHPA delineation.
Human activities, whether agricultural, industrial, commercial, or domestic, can contribute to ground water quality deterioration. In order to protect the ground water exploited by a production well, it is essential to develop a good knowledge of the flow system and to adequately delineate the area surrounding the well within which potential contamination sources should be managed. Many methods have been developed to delineate such a wellhead protection area (WHPA). The integration of more information on the geologic and hydrogeologic characteristics of the study area increases the precision of any given WHPA delineation method. From a practical point of view, the WHPA delineation methods allowing the simplest and least expensive integration of the available information should be favored. This paper presents a comparative study in which nine different WHPA delineation methods were applied to a well and a spring in an unconfined granular aquifer and to a well in a confined highly fractured rock aquifer. These methods range from simple approaches to complex computer models. Hydrogeological mapping and numerical modeling with MODFLOW-MODPATH were used as reference methods to respectively compare the delineation of the zone of contribution and the zone of travel obtained from the various WHPA methods. Although applied to simple ground water flow systems, these methods provided a relatively wide range of results. To allow a realistic delineation of the WHPA in aquifers of variable geometry, a WHPA delineation method should ensure a water balance and include observed or calculated regional flow characteristics.


Epidemiological approaches to disability in the elderly: from measurement to the projection of need.
Measures of disability, in the elderly as a proxy for autonomy, are essential for assessment of physical and mental wellbeing, estimates of needs for care of individuals and communities and for the planning of services. Such measures need to be combined with clinical information to complete the diagnosis for the individual. For groups, assessments are subject to the interactions of reported disability with age, social class and culture, even when using standardized questionnaires. These intervening variables, as well as the representatives of the groups sampled, must be considered when extrapolating the findings to the whole population and, particularly, in the forecasting of future disability patterns. Translation of disability measurements into needs for service, now and in the future, is a complex procedure further confounded by social expectations and professional judgement.


Evidence for the role of selection for reproductively advantageous alleles in human aging.
The antagonistic pleiotropy hypothesis posits that natural selection for pleiotropic mutations that confer earlier or more reproduction but impair the post-reproductive life causes aging. This hypothesis of the evolutionary origin of aging is supported by case studies but lacks unambiguous genomic evidence. Here, we genomically test this hypothesis using the genotypes, reproductive phenotypes, and death registry of 276,406 U.K. Biobank participants. We observe a strong, negative genetic correlation between reproductive traits and life span. Individuals with higher polygenetic scores for reproduction (PGSR) have lower survivorships to age 76 (SV76), and PGSR increased over birth cohorts from 1940 to 1969. Similar trends are seen from individual genetic variants examined. The antagonistically pleiotropic variants are often associated with cis-regulatory effects across multiple tissues or on multiple target genes. These and other findings support the antagonistic pleiotropy hypothesis of aging in humans and point to potential molecular mechanisms of the reproduction-life-span antagonistic pleiotropy.


Climate change, extreme events, and increased risk of salmonellosis: foodborne diseases active surveillance network (FoodNet), 2004-2014.
Infections with nontyphoidal Salmonella cause an estimated 19,336 hospitalizations each year in the United States. Sources of infection can vary by state and include animal and plant-based foods, as well as environmental reservoirs. Several studies have recognized the importance of increased ambient temperature and precipitation in the spread and persistence of Salmonella in soil and food. However, the impact of extreme weather events on Salmonella infection rates among the most prevalent serovars, has not been fully evaluated across distinct U.S. regions. To address this knowledge gap, we obtained Salmonella case data for S. Enteriditis, S. Typhimurium, S. Newport, and S. Javiana (2004-2014; n = 32,951) from the Foodborne Diseases Active Surveillance Network (FoodNet), and weather data from the National Climatic Data Center (1960-2014). Extreme heat and precipitation events for the study period (2004-2014) were identified using location and calendar day specific 95th percentile thresholds derived using a 30-year baseline (1960-1989). Negative binomial generalized estimating equations were used to evaluate the association between exposure to extreme events and salmonellosis rates. We observed that extreme heat exposure was associated with increased rates of infection with S. Newport in Maryland (Incidence Rate Ratio (IRR): 1.07, 95% Confidence Interval (CI): 1.01, 1.14), and Tennessee (IRR: 1.06, 95% CI: 1.04, 1.09), both FoodNet sites with high densities of animal feeding operations (e.g., broiler chickens and cattle). Extreme precipitation events were also associated with increased rates of S. Javiana infections, by 22% in Connecticut (IRR: 1.22, 95% CI: 1.10, 1.35) and by 5% in Georgia (IRR: 1.05, 95% CI: 1.01, 1.08), respectively. In addition, there was an 11% (IRR: 1.11, 95% CI: 1.04-1.18) increased rate of S. Newport infections in Maryland associated with extreme precipitation events. Overall, our study suggests a stronger association between extreme precipitation events, compared to extreme heat, and salmonellosis across multiple U.S. regions. In addition, the rates of infection with Salmonella serovars that persist in environmental or plant-based reservoirs, such as S. Javiana and S. Newport, appear to be of particular significance regarding increased heat and rainfall events.


Interleukin-28B genetic variations and spontaneous clearance of hepatitis C antibody-positive blood donors in China.
Single-nucleotide polymorphisms (SNPs) near the interleukin (IL)28B gene have been shown to be associated with spontaneous clearance of hepatitis C virus (HCV) infection in western populations. Anti-HCV-reactive and HCV RNA-negative samples were often found in Chinese blood donors. This study was to determine whether the IL28B gene variant is also associated with the natural outcome of HCV infection in Chinese blood donors. A total of 111 anti-HCV-negative blood donors and 100 anti-HCV-reactive subjects were enrolled in this study. HCV-infected blood donors were classified into spontaneous clearance group and chronic infection group through HCV RNA detection. IL28B genetic variations (rs8099917 and rs12979860) were genotyped by polymerase chain reaction and direct sequencing. A total of 24 (24.0%) donors spontaneously cleared HCV. The rs12979860 C allele frequency was 90.3% and the rs8099917 T allele frequency was 96.7% in our study. The rs12979860 CC genotype was strongly associated with spontaneous HCV clearance. Female blood donors had a higher rate of spontaneous HCV clearance than male donors (15/42 [35.7%] vs. 9/58 [15.5%], p = 0.02). The rs12979860 CC variant upstream of IL28B gene is associated with spontaneous clearance of HCV in Chinese blood donors and female donors are more likely to clear HCV infection than male donors.


Quality of life improvement in patients treated with degarelix versus leuprorelin for advanced prostate cancer.
We used responses to questionnaires included in the CS21 degarelix trial and published mapping algorithms to address the paucity of evidence for health related quality of life in patients with advanced hormone dependent prostate cancer treated with degarelix. We measured health related quality of life in 610 patients enrolled in the CS21 trial using SF-12® and EORTC QLQ-C30. Based on responses to these questionnaires we estimated patient utility using 4 published mapping algorithms. Utility was tested for relationships with aspects of the symptom and side effect burden that may be affected by degarelix treatment, that is prostate specific antigen progression and adverse events. Average utility in patients without prostate specific antigen progression or an adverse event was 0.742, similar to previously published utilities for nonprogressed prostate cancer states. Prostate specific antigen progression was associated with a utility decrement of between 0.062 and 0.134 depending on the mapping algorithm used. Of adverse events considered in our analysis musculoskeletal events were associated with the greatest effects on patient utility with a decrement of between 0.029 and 0.086. The 4 mapping algorithms generated similar utility estimates, although values derived from SF-12 were consistently lower than those derived from EORTC QLQ-C30. Prostate specific antigen progression status and the incidence of treatment and disease related adverse events result in significant decrements to patient health related quality of life. By slowing prostate specific antigen progression degarelix may improve patient utility and the health related quality of life burden.


The Korean prediction model for adolescents' future smoking intentions.
The purpose of this study was to develop a prediction model for future smoking intention among Korean adolescents aged 13 to 15 in order to identify the high risk group exposed to future smoking. The data was collected from a total of 5940 students who participated in a self-administrated questionnaire of a cross-sectional school-based survey, the 2004 Korea Global Youth Tobacco Survey. Chi-square tests and logistic regression analyses were carried out to identify the relevant determinants associated with intentions of adolescents' future smoking. Receiver Operation Characteristic (ROC) assessment was applied to evaluate the explanation level of the developed prediction model. 8.4% of male and 7.2% of female participants show their intentions of future smoking. Among non-smoking adolescents; who have past smoking experience [odds ratio (OR) 2.73; 95% confidence interval (CI) 1.92 - 3.88]; who have intentions of smoking when close friends offer a cigarette (OR 31.47; 95% CI = 21.50 - 46.05); and who have friends that are mostly smokers (OR 5.27; 95% CI = 2.85 - 9.74) are more likely to be smokers in the future. The prediction model developed from this study consists of five determinants; past smoking experience; parents smoking status; friends smoking status; ownership of a product with a cigarette brand logo; and intentions of smoking from close friends' cigarette offer. The area under the ROC curve was 0.8744 (95% CI=0.85 - 0.90) for current non-smokers. For efficiency, school-based smoking prevention programs need to be designed to target the high risk group exposed to future smoking through the prediction model developed by the study, instead of implementing the programs for all the students.


Single-Cell Analysis Using Drop-on-Demand Inkjet Printing and Probe Electrospray Ionization Mass Spectrometry.
This study describes a novel method for single-cell analysis and lipid profiling by combining drop-on-demand inkjet cell printing and probe electrospray ionization mass spectrometry (PESI-MS). Through inkjet sampling of a cell suspension, droplets with single cells were generated, precisely dripped onto a tungsten-made electrospray ionization needle, and immediately sprayed under a high-voltage electric field. Lipid fingerprints of single cells were obtained by a mass spectrometry (MS) detector. A homemade magnetic stirring device was applied to the cell suspension reservoir, which controlled the homogeneous distribution of cells in liquid and improved the single-cell-droplet percentage by 43.8%. Eight types of single cells were screened in our platform and further differentiated by principal component analysis based on cellular surface phospholipids. Thus, this study successfully provides a facile method for the direct MS profiling of single-cell lipids by PESI-MS.


PDZK1 is a novel factor in breast cancer that is indirectly regulated by estrogen through IGF-1R and promotes estrogen-mediated growth.
Although a relationship between PDZK1 expression and estrogen receptor (ER)-α stimulation has been suggested, the nature of such a connection and the function of PDZK1 in breast cancer remain unknown. Human tissue microarrays (cancer tissue: 262 cores; normal tissue: 87 cores) and breast cancer cell lines were used to conduct the study. We show that PDZK1 protein expression is tightly correlated with human breast malignancy, is negatively correlated with age and had no significant correlation with ER-α expression levels. PDZK1 exhibited an exclusive epithelial expression with mostly cytosolic subcellular localization. Additionally, 17β-estradiol induced PDZK1 expression above its basal level more than 24 h after treatment in MCF-7 cells. PDZK1 expression was indirectly regulated by ER-α stimulation, requiring insulinlike growth factor 1 receptor (IGF-1R) expression and function. The molecular link between PDZK1 and IGF-1R was supported by a significant correlation between protein and mRNA levels (r = 0.591, p < 0.001, and r = 0.537, p < 0.001, respectively) of the two factors in two different cohorts of human breast cancer tissues. Interestingly, PDZK1 knockdown in MCF-7 cells blocked ER-dependent growth and reduced c-Myc expression, whereas ectopic expression of PDZK1 enhanced cell proliferation in the presence or absence of 17β-estradiol potentially through an increase in c-Myc expression, suggesting that PDZK1 has oncogenic activity. PDKZ1 also appeared to interact with the Src/ER-α/epidermal growth factor receptor (EGFR) complex, but not with IGF-1R and enhanced EGFR-stimulated MEK/ERK1/2 signaling. Collectively, our results clarify the relationship between ER-α and PDZK1, propose a direct relationship between PDZK1 and IGF-1R, and identify a novel oncogenic activity for PDZK1 in breast cancer.


[Species makeup of coagulase-negative staphylococci of clinical origin].
The classification of Kloos--Schleifer was used for the identification of 175 clinical strains of coagulase-negative staphylococci. The species of staphylococci in 50.3% of the cultures were determined; as a result, S. epidermidis and S. saprophyticus were not found to be in the majority among them. The authors concluded that the above-mentioned scheme, as well as its short variant proposed by the International Subcommittee on the Taxonomy of Staphylococci and Micrococci could not be recommended for practical use.


Use of a structural analogue versus a stable isotope labeled internal standard for the quantification of angiotensin IV in rat brain dialysates using nano-liquid chromatography/tandem mass spectrometry.
Quantifying low concentrations of neuropeptides in microdialysates requires a selective and sensitive analysis technique, such as nano-liquid chromatography/electrospray ionization tandem mass spectrometry (nanoLC/ESI-MS/MS). However, we observed reduced accuracy of the method due to matrix effects. Indeed, ESI-MS detection is known to be sensitive to matrix effects. Moreover, dialysates are complex mixtures of small molecules, peptides and other matrix compounds that can influence the ionization efficiency of the neuropeptide of interest and the stability of the peptide in the samples. In the study reported in this paper, we investigated whether the use of an internal standard (IS) can correct for these matrix effects. As a model compound for neuropeptides we used angiotensin IV (Ang IV). We compared the use of a structural analogue (norleucine1-Ang IV) with a stable isotope labeled (SIL) analogue. Linearity of the method was improved when either of the proposed ISs were applied. Only when using the SIL-IS could the repeatability of injection and the method's precision and accuracy be improved. Finally, the IS was able to correct for degradation of Ang IV in dialysates, prolonging the possible storage period of the samples. We conclude that the structural analogue is not suited as an IS and that the application of a SIL analogue is indispensable when quantifying Ang IV in dialysates using nanoLC/ESI-MS/MS detection.


Trauma Center Youth Violence Screening and Brief Interventions: A Multisite Pilot Feasibility Study.
Every day, 16 American youths between the ages of 10 and 24 years are murdered; 84% of these fatalities involve a firearm. Nearly half of traumatic youth deaths result from violence-related injuries. In 2013, 580,250 youth suffered nonfatal, assault-related injuries, necessitating emergency department treatment. The aim of this multisite pilot study was to examine the process, feasibility, and challenges of violence brief interventions (VBIs). The participants were youth between 15 and 25 years of age, at 2 major Level 1 trauma centers (TCs; TC1, TC2) in the Southeastern United States. Eligible participants (N = 38; TC1: n = 20, TC2: n = 18) received at least 1 VBI during their hospital stay, which provided information about individual screening results and elicited patients' perspectives on violent and risky behaviors. More participants at TC2 than at TC1 completed 2 VBI sessions. Barriers to and support of implementation were identified at both sites, and factors for improving implementation were identified, including the need for staff support through clinical guidelines and coordinated prevention and outreach programs. Further research is needed to identify factors for successful implementation of VBIs in TCs.


Graft failure following allogeneic blood and marrow transplant: evidence-based nursing case study review.
Despite the advances made since the earliest days of transplant therapy, graft failure following allogeneic blood and marrow transplant is still a life-threatening complication. This article reviews the science of graft failure and uses a case study presentation to address how an oncology nursing staff was motivated by a patient's experience of graft failure. An evidence-based literature review was undertaken to answer three relevant clinical questions: (a) What factors contribute to graft failure in patients receiving allogeneic hematopoietic stem cell transplants? (b) What interventions are appropriate for these patients? and (c) How can this information assist nursing staff in providing improved care for these patients? An example of the table of evidence is provided.


Treatment with 670 nm light up regulates cytochrome C oxidase expression and reduces inflammation in an age-related macular degeneration model.
Inflammation is an umbrella feature of ageing. It is present in the aged retina and many retinal diseases including age-related macular degeneration (AMD). In ageing and in AMD mitochondrial function declines. In normal ageing this can be manipulated by brief exposure to 670 nm light on the retina, which increases mitochondrial membrane potential and reduces inflammation. Here we ask if 670 nm exposure has the same ability in an aged mouse model of AMD, the complement factor H knockout (CFH(-/-)) where inflammation is a key feature. Further, we ask whether this occurs when 670 nm is delivered briefly in environmental lighting rather than directly focussed on the retina. Mice were exposed to 670 nm for 6 minutes twice a day for 14 days in the form of supplemented environmental light. Exposed animals had significant increase in cytochrome c oxidase (COX), which is a mitochondrial enzyme regulating oxidative phosphorylation.There was a significant reduction in complement component C3, an inflammatory marker in the outer retina. Vimetin and glial fibrillary acidic protein (GFAP) expression, which reflect retinal stress in Muller glia, were also significantly down regulated. There were also significant changes in outer retinal macrophage morphology. However, amyloid beta (Aβ) load, which also increases with age in the outer retina and is pro-inflammatory, did not change. Hence, 670 nm is effective in reducing inflammation probably via COX activation in mice with a genotype similar to that in 50% of AMD patients even when brief exposures are delivered via environmental lighting. Further, inflammation can be reduced independent of Aβ. The efficacy revealed here supports current early stage clinical trials of 670 nm in AMD patients.


Strain and sex matters: Differences in nicotine self-administration between outbred and recombinase-driver transgenic rat lines.
Preclinical studies of nicotine self-administration provide important value for the field as they are highly rigorous, controlled, can be conducted quickly, and are generalizable to humans. Given the translational value of the nicotine self-administration model, and the relatively new guidelines of the National Institutes of Health to include sex as a biological variable, strain and sex differences in nicotine acquisition were examined here in two outbred rat strains. Sprague-Dawley (SD) and Long-Evans (LE; wildtype and cholinergic acetyltransferase cre-recombinase transgenic) rats of each sex were implanted with indwelling intravenous jugular catheters. Rats were trained to self-administer nicotine (0.02 mg/kg per infusion, paired with contingent light + tone stimuli). Acquisition criteria were set at a minimum active:inactive response ratio of 2:1 and a minimum of 10 infusions per session, both of which had to be met for a minimum of 10 sessions. Across 10 sessions, male SD rats self-administered significantly more nicotine than female SD rats (p < .05), indicating a sex difference in this strain. LE females self-administered more nicotine than SD females indicative of a strain difference between females (p < .05). SD males increased nicotine infusions across sessions compared to LE males and SD females (p < .05). No strain or sex differences were observed in the number of sessions to reach criteria. No differences between wildtype and transgenic LE rats were observed. These results demonstrate sex and strain differences in nicotine self-administration between SD and LE rats and may lend insight into development of other nicotine self-administration models, where sex and strain may impact acquisition. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


Automated glycopeptide analysis--review of current state and future directions.
Glycosylation of proteins is involved in immune defense, cell-cell adhesion, cellular recognition and pathogen binding and is one of the most common and complex post-translational modifications. Science is still struggling to assign detailed mechanisms and functions to this form of conjugation. Even the structural analysis of glycoproteins-glycoproteomics-remains in its infancy due to the scarcity of high-throughput analytical platforms capable of determining glycopeptide composition and structure, especially platforms for complex biological mixtures. Glycopeptide composition and structure can be determined with high mass-accuracy mass spectrometry, particularly when combined with chromatographic separation, but the sheer volume of generated data necessitates computational software for interpretation. This review discusses the current state of glycopeptide assignment software-advances made to date and issues that remain to be addressed. The various software and algorithms developed so far provide important insights into glycoproteomics. However, there is currently no freely available software that can analyze spectral data in batch and unambiguously determine glycopeptide compositions for N- and O-linked glycopeptides from relevant biological sources such as human milk and serum. Few programs are capable of aiding in structural determination of the glycan component. To significantly advance the field of glycoproteomics, analytical software and algorithms are required that: (i) solve for both N- and O-linked glycopeptide compositions, structures and glycosites in biological mixtures; (ii) are high-throughput and process data in batches; (iii) can interpret mass spectral data from a variety of sources and (iv) are open source and freely available.


Interaction of hyaluronate with the surface of simian virus 40-transformed 3T3 cells: aggregation and binding studies.
Previous studies have shown that the divalent-cation independent aggregation of simian virus 40-transformed 3T3 cells (SV-3T3) is mediated by the interaction of endogenous hyaluronate with binding sites on the cell surface. In the present study, the nature of the interaction of hyaluronate with the binding sites was characterized further by examining both the aggregation of SV-3T3 cells and the binding of exogenously added [3H]hyaluronate. The aggregation of SV-3T3 cells was found to be: (1) optimal at pH 7; (2) only slightly affected by variations in temperature between 0 and 40 degrees C; (3) more potently inhibited by the addition of high molecular weight preparations of hyaluronate than by lower molecular weight preparations; and (4) enhanced by increasing the ionic strength of the medium. With the exception of the effects of temperature, these characteristics correspond closely to those previously described for binding of [3H]hyaluronate. The effect of ionic strength on the binding of [3H]hyaluronate by SV-3T3 cells was found to correspond closely to its effect on lowering the viscosity of hyaluronate. Both effects (i.e. binding and reduction in viscosity) had similar dose-response curves for NaCl and MgCl2. This close correspondence between binding and the reduction of viscosity suggested that the effect of ionic strength on binding was due to a change in the structure of hyaluronate. Lowering the temperature (40 to 0 degrees C) also enhanced the binding of [3H]hyaluronate to SV-3T3 cells. However, this effect was blocked if the cells were first fixed with glutaraldehyde, suggesting that temperature mediates its effect through the binding sites. Additional experiments showed that the binding of [3H]hyaluronate by SV-3T3 cells could be prevented by preincubating the cells with several types of lectins.


Cataract surgery in patients with age-related macular degeneration: one-year outcomes.
To determine whether patients with age-related macular degeneration (ARMD) benefit from cataract surgery and to assess the risk of progression of preexisting maculopathy 4 and 12 months postoperatively. Princess Alexandra Eye Pavilion, Royal Infirmary of Edinburgh, Edinburgh, Scotland. Two groups of patients were evaluated prospectively. The study group comprised patients with ARMD scheduled to have cataract surgery (n = 40). The control group comprised patients with ARMD not having cataract surgery (n = 43). Patients were assessed at baseline (preoperatively) and 4 and 12 months postoperatively. Assessment included visual function tests and quality of life (QoL) measures. The mean values for each item tested were obtained for each group at each visit, and comparisons between visits were done using the Wilcoxon signed rank test. There were significant benefits of cataract surgery in terms of visual function and QoL measures at 4 and 12 months. There was no increased risk of progression of maculopathy in the study group. There were no significant differences in the items tested in the control group. One year postoperatively, QoL benefits were maintained in the study group and there was no increased risk of progression of maculopathy in patients with mild and moderate degrees of ARMD. Larger numbers of patients must be assessed prospectively for longer periods to determine the relative risk of progression of different stages of ARMD after cataract surgery.


Formulating policies for the welfare of animals during long distance transportation.
Long distance transportation of animals creates much public concern in the European Union (EU) and elsewhere, partly because of its visibility to the general public. The protection of animals during transport is thus a key element of EU policy for farm animal welfare. At the same time, animal transport has been vital to the structure of the food chain in Europe since the beginning of the Common Agricultural Policy. The authors describe the formulation of EU policy on long distance transportation. Initiatives are based on scientific risk assessment and considerations of international guidelines. The two main objectives of EU policy are to reduce long distance transportation as far as possible and to upgrade standards for transported animals. The extent of detail in regulations depends on the ability of the sector concerned to address issues and on continual upgrading of the awareness and knowledge of transport operators on animal welfare which is universally important for progress. The economic impact of legislative measures must be evaluated as part of the policy process, noting that proper animal welfare standards can generate direct and indirect economic advantages. Awareness of these welfare advantages in all sectors is essential for raising the quality of enforcement. Finally, policy goals should be monitored to verify the extent of their fulfilment. Efforts from competent authorities and transport companies in Europe are improving the situation. However, a strong legislative framework is likely to remain the best option for the coming years to ensure that the welfare of transported animals is more than just a minimum.


Covalent trapping of methyllycaconitine at the α4-α4 interface of the α4β2 nicotinic acetylcholine receptor: antagonist binding site and mode of receptor inhibition revealed.
The α4β2 nicotinic acetylcholine receptors (nAChRs) are widely expressed in the brain and are implicated in a variety of physiological processes. There are two stoichiometries of the α4β2 nAChR, (α4)2(β2)3 and (α4)3(β2)2, with different sensitivities to acetylcholine (ACh), but their pharmacological profiles are not fully understood. Methyllycaconitine (MLA) is known to be an antagonist of nAChRs. Using the two-electrode voltage clamp technique and α4β2 nAChRs in the Xenopus oocyte expression system, we demonstrate that inhibition by MLA occurs via two different mechanisms; that is, a direct competitive antagonism and an apparently insurmountable mechanism that only occurs after preincubation with MLA. We hypothesized an additional MLA binding site in the α4-α4 interface that is unique to this stoichiometry. To prove this, we covalently trapped a cysteine-reactive MLA analog at an α4β2 receptor containing an α4(D204C) mutation predicted by homology modeling to be within reach of the reactive probe. We demonstrate that covalent trapping results in irreversible reduction of ACh-elicited currents in the (α4)3(β2)2 stoichiometry, indicating that MLA binds to the α4-α4 interface of the (α4)3(β2)2 and providing direct evidence of ligand binding to the α4-α4 interface. Consistent with other studies, we propose that the α4-α4 interface is a structural target for potential therapeutics that modulate (α4)3(β2)2 nAChRs.


Current Utilization and Perceptions of Formal Education of Point-Of-Care-Ultrasound in Urology.
To assess the extent of formal POCUS training, current utilization of POCUS, and contemporary perceptions of POCUS amongst urologists. A survey including questions regarding demographics, prior ultrasound education, current ultrasound utilization in practice/training, perceived optimal POCUS utilization, and the perception of formal ultrasound training was developed. The survey was disseminated to residency program directors (PDs) via the SAU and members of AUA sub-section organizations. Data was collected via Redcap. A total of 40 PDs and 159 other respondents completed the survey with approximately half (51%) in an academic practice and two-thirds (68%) with more than 10 years in practice. PD response rate was 28%, and general response rate was 2%. Among all respondents, 95% (186/196) and 82% (160/194) agreed/strongly agreed formal POCUS training would be worthwhile during and after residency. Among urology residency program directors, 93% (37/40) agreed/strongly agreed that formal POCUS training is worthwhile in residency. The majority of respondents used some form of ultrasound in practice (77%, 154/199). However, only 37% (72/199) of all respondents had prior formal POCUS training, and 19% (5/26) of program directors reported formal training in their programs. Of respondents without formal training, 63% (80/127) reported interest in pursuing formal training. POCUS is widely utilized in many practices. Yet, most urologists have not participated in formal POCUS training and most programs do not have curricula. Urologists have favorable opinions of the utility, safety, and efficacy of POCUS and desire training.


Bone mineral density, osteoporosis, and osteoporotic fractures: a genome-wide association study.
Osteoporosis is diagnosed by the measurement of bone mineral density, which is a highly heritable and multifactorial trait. We aimed to identify genetic loci that are associated with bone mineral density. In this genome-wide association study, we identified the most promising of 314 075 single nucleotide polymorphisms (SNPs) in 2094 women in a UK study. We then tested these SNPs for replication in 6463 people from three other cohorts in western Europe. We also investigated allelic expression in lymphoblast cell lines. We tested the association between the replicated SNPs and osteoporotic fractures with data from two studies. We identified genome-wide evidence for an association between bone mineral density and two SNPs (p<5x10(-8)). The SNPs were rs4355801, on chromosome 8, near to the TNFRSF11B (osteoprotegerin) gene, and rs3736228, on chromosome 11 in the LRP5 (lipoprotein-receptor-related protein) gene. A non-synonymous SNP in the LRP5 gene was associated with decreased bone mineral density (rs3736228, p=6.3x10(-12) for lumbar spine and p=1.9x10(-4) for femoral neck) and an increased risk of both osteoporotic fractures (odds ratio [OR] 1.3, 95% CI 1.09-1.52, p=0.002) and osteoporosis (OR 1.3, 1.08-1.63, p=0.008). Three SNPs near the TNFRSF11B gene were associated with decreased bone mineral density (top SNP, rs4355801: p=7.6x10(-10) for lumbar spine and p=3.3x10(-8) for femoral neck) and increased risk of osteoporosis (OR 1.2, 95% CI 1.01-1.42, p=0.038). For carriers of the risk allele at rs4355801, expression of TNFRSF11B in lymphoblast cell lines was halved (p=3.0x10(-6)). 1883 (22%) of 8557 people were at least heterozygous for these risk alleles, and these alleles had a cumulative association with bone mineral density (trend p=2.3x10(-17)). The presence of both risk alleles increased the risk of osteoporotic fractures (OR 1.3, 1.08-1.63, p=0.006) and this effect was independent of bone mineral density. Two gene variants of key biological proteins increase the risk of osteoporosis and osteoporotic fracture. The combined effect of these risk alleles on fractures is similar to that of most well-replicated environmental risk factors, and they are present in more than one in five white people, suggesting a potential role in screening.


Salmonella enterica serovar Typhimurium typing by prophage-specific PCR.
Recent data from microarray analysis have shown that integrated prophages are the most frequent sources of genomic variation between different strains of Salmonella enterica serovar Typhimurium (S. Typhimurium). This led us to hypothesize that PCR detection of the integrated prophages might be an efficient typing tool that could be used as an alternative to PFGE. In this study, we optimized four triplex PCRs specific for 12 target sequences of mostly prophage origin, and tested them in 102 field strains. The same set of strains was also characterized by PFGE. Among the strains, 22 different multiplex PCR, and 25 different PFGE profiles, were identified. Despite the fact that the PFGE was slightly more discriminatory, multiplex PCR typing, owing to its simplicity and potential of simple data sharing between laboratories, represents an interesting user-friendly alternative to PFGE typing of S. Typhimurium.


Stimulation of calcitonin gene-related peptide synthesis and release: mechanisms for a novel antihypertensive drug, rutaecarpine.
Previous investigations have demonstrated that capsaicin-sensitive primary sensory nerves play an important role in modulation of the peripheral resistance of the circulation system. The vanilloid receptor subtype 1 (VR1) is expressed almost exclusively in the primary sensory nerves and cell bodies of these sensory neurons. Rutaecarpine (Rut) can relax vascular smooth muscle via stimulation of calcitonin gene-related peptide (CGRP) release by activation of VR1. In the present study, we examined the depressor effect of Rut and the possible mechanisms in the phenol-induced hypertensive rats, in which hypertension was induced by injecting 50 microl of 10% phenol in the lower pole of the left kidney. Acute administration of Rut (30, 100 or 300 microg/kg, i.v.) caused a depressor effect concomitantly with an increase in the plasma concentration of CGRP in a dose-dependent manner, which was blocked by capsaicin (used to deplete the CGRP from sensory nerves) or capsazepine (a competitive VR1 antagonist), causing an approximately 85% and approximately 80% change in mean arterial pressure, respectively, and by either of them, causing an approximately 90% elevation of plasma CGRP. In the chronic study, Rut at a dose of 3 or 6 mg/kg per day significantly lowered tail-cuff systolic blood pressure to 159 +/- 8 and 136 +/- 10 mmHg, respectively, compared with hypertensive rats (179 +/- 8 mmHg), and caused a sustained hypotensive effect from day 6 on. Pretreatment with capsaicin blocked the depressor effect of Rut by approximately 65%. Treatment with Rut significantly increased the synthesis and release of CGRP, as shown by the increase in the levels of CGRP mRNA and peptide in the dorsal root ganglia, the density of CGRP immunoreactive nerve fibers in the mesenteric artery, the CGRP content in the spinal cord and the plasma concentration of CGRP, which was markedly attenuated by pretreatment with capsaicin. These results suggest, for the first time, that the hypotensive effect of Rut is mediated by stimulation of CGRP synthesis and release via activation of VR1 in the phenol-induced hypertensive rat.


Update: severe respiratory illness associated with a novel coronavirus--worldwide, 2012-2013.
CDC continues to work closely with the World Health Organization (WHO) and other partners to better understand the public health risk posed by a novel coronavirus that was first reported to cause human infection in September 2012. Genetic sequence analyses have shown that this new virus is different from any other known human coronaviruses, including the one that caused severe acute respiratory syndrome (SARS). As of March 7, 2013, a total of 14 confirmed cases of novel coronavirus infection have been reported to WHO, with eight deaths. Illness onsets have occurred from April 2012 through February 2013. To date, no cases have been reported in the United States.


Medical Costs Among Youth Younger Than 20 Years of Age With and Without Diabetic Ketoacidosis at the Time of Diabetes Diagnosis.
While diabetic ketoacidosis (DKA) is common in youth at the onset of the diabetes, the excess costs associated with DKA are unknown. We aimed to quantify the health care services use and medical care costs related to the presence of DKA at diagnosis of diabetes. We analyzed data from the U.S. MarketScan claims database for 4,988 enrollees aged 3-19 years insured in private fee-for-service plans and newly diagnosed with diabetes during 2010-2016. Youth with and without DKA at diabetes diagnosis were compared for mean health care service use (outpatient, office, emergency room, and inpatient visits) and medical costs (outpatient, inpatient, prescription drugs, and total) for 60 days prior to and 60 days after diabetes diagnosis. A two-part model using generalized linear regression and logistic regression was used to estimate medical costs, controlling for age, sex, rurality, health plan, year, presence of hypoglycemia, and chronic pulmonary condition. All costs were adjusted to 2016 dollars. At diabetes diagnosis, 42% of youth had DKA. In the 60 days prior to diabetes diagnosis, youth with DKA at diagnosis had less health services usage (e.g., number of outpatient visits: -1.17; P < 0.001) and lower total medical costs (-$635; P < 0.001) compared with youth without DKA at diagnosis. In the 60 days after diagnosis, youth with DKA had significantly greater health care services use and health care costs ($6,522) compared with those without DKA. Among youth with newly diagnosed diabetes, DKA at diagnosis is associated with significantly higher use of health care services and medical costs.


Protein release from galactoglucomannan hydrogels: influence of substitutions and enzymatic hydrolysis by beta-mannanase.
O-Acetyl-galactoglucomannan (AcGGM) is the major soft-wood hemicellulose. Structurally modified AcGGM and hydrogels of AcGGM were prepared. The degree of substitution (DS) of AcGGM was modified enzymatically with alpha-galactosidase, and chemically with an acrylate derivative, 2-hydroxyethylmethacrylate (HEMA). The hydrolysis of AcGGM with beta-mannanase was shown to increase with decreasing DS. AcGGM hydrogels were prepared from chemically modified AcGGM with varying DS of HEMA. Bovine serum albumin (BSA) was encapsulated in hydrogels. A spontaneous burst release of BSA was decreased with increased DS of HEMA. The addition of beta-mannanase significantly enhanced the BSA release from hydrogels with a DS of 0.36, reaching a maximum of 95% released BSA after eight hours compared to 60% without enzyme. Thus, both the pendant group composition and the enzyme action are valuable tools in the tailoring of hydrogel release profiles of potential interest for intestine drug delivery.


[Leptin and adiponectin in pathogenesis of metabolic syndrome].
Metabolic syndrome, hypernutrition syndrome is a phylogenetically determined sequence of symptoms having common pathogenesis. Etiologically, it is due to excessive intake of adequate food. Enterocytes and omental adipose cells form an early phylogenetically unified and paracrinically regulated community that alternately realize exo- and endotrophic biological reactions. Visceral obesity high level of non-esterified fatty acids (FA), formation of plasma FA pool and their integration into the endothelial membrane, and enlargement of enterocytes are the main cause of increased hydrodynamic pressure. Toll-like receptors perceive albumin associates containing a supraphysiological number of FA as aliens and initiate the inflammatory reaction. Lipid-overloaded cells undergo "endoplasmic stress", abnormal protein synthesis (folding), and apoptosis-like death. Visceral fat serves as phylogenetically early FA depot for the realization of biological functions of homeostasis, trophology endoecology, and adaptation; it is anatomically confined and regulated at the level of paracrine communities. The subcutaneous depot realizes the phylogenetically late locomotor function; its size is not anatomically restricted. Visceral adipose cells have no receptors for phylogenetically late insulin (INS). Cells of the subcutaneous FA depot are specialized adipocytes having INS and GLUT4 receptors. They are regulated by the phylogenetically late humoral mechanisms at the total body level. Leptin initiates in vivo humoral, hypothalamic regulation of the size of INS-insensitive visceral adipose cells quantitatively programmed in ontogenesis; also, it prevents "endoplasmic stress" and apoptosis, regulates food consumption. Leptin initiates switching FA storage from the visceral adipocyte pool to the subcutaneous one. Adiponectin is a phylogenetically late humoral inducer of regulation of the optimal in vivo number of cells from the hypothalamus level. It is biologically predetermined to regulate the number (proliferation) of INS-dependent adipocytes in subcutaneous fat.


Remote mutations and active site dynamics correlate with catalytic properties of purine nucleoside phosphorylase.
It has been found that with mutation of two surface residues (Lys(22) --> Glu and His(104) --> Arg) in human purine nucleoside phosphorylase (hPNP), there is an enhancement of catalytic activity in the chemical step. This is true although the mutations are quite remote from the active site, and there are no significant changes in crystallographic structure between the wild-type and mutant active sites. We propose that dynamic coupling from the remote residues to the catalytic site may play a role in catalysis, and it is this alteration in dynamics that causes an increase in the chemical step rate. Computational results indicate that the mutant exhibits stronger coupling between promotion of vibrations and the reaction coordinate than that found in native hPNP. Power spectra comparing native and mutant proteins show a correlation between the vibrations of Immucillin-G (ImmG):O5'...ImmG:N4' and H257:Ndelta...ImmG:O5' consistent with a coupling of these motions. These modes are linked to the protein promoting vibrations. Stronger coupling of motions to the reaction coordinate increases the probability of reaching the transition state and thus lowers the activation free energy. This motion has been shown to contribute to catalysis. Coincident with the approach to the transition state, the sum of the distances of ImmG:O4'...ImmG:O5'...H257:Ndelta became smaller, stabilizing the oxacarbenium ion formed at the transition state. Combined results from crystallography, mutational analysis, chemical kinetics, and computational analysis are consistent with dynamic compression playing a significant role in forming the transition state. Stronger coupling of these pairs is observed in the catalytically enhanced mutant enzyme. That motion and catalysis are enhanced by mutations remote from the catalytic site implicates dynamic coupling through the protein architecture as a component of catalysis in hPNP.


Role of serotonin via 5-HT2B receptors in the reinforcing effects of MDMA in mice.
The amphetamine derivative 3,4-methylenedioxymethamphetamine (MDMA, ecstasy) reverses dopamine and serotonin transporters to produce efflux of dopamine and serotonin, respectively, in regions of the brain that have been implicated in reward. However, the role of serotonin/dopamine interactions in the behavioral effects of MDMA remains unclear. We previously showed that MDMA-induced locomotion, serotonin and dopamine release are 5-HT(2B) receptor-dependent. The aim of the present study was to determine the contribution of serotonin and 5-HT(2B) receptors to the reinforcing properties of MDMA.We show here that 5-HT(2B) (-/-) mice do not exhibit behavioral sensitization or conditioned place preference following MDMA (10 mg/kg) injections. In addition, MDMA-induced reinstatement of conditioned place preference after extinction and locomotor sensitization development are each abolished by a 5-HT(2B) receptor antagonist (RS127445) in wild type mice. Accordingly, MDMA-induced dopamine D1 receptor-dependent phosphorylation of extracellular regulated kinase in nucleus accumbens is abolished in mice lacking functional 5-HT(2B) receptors. Nevertheless, high doses (30 mg/kg) of MDMA induce dopamine-dependent but serotonin and 5-HT(2B) receptor-independent behavioral effects.These results underpin the importance of 5-HT(2B) receptors in the reinforcing properties of MDMA and illustrate the importance of dose-dependent effects of MDMA on serotonin/dopamine interactions.


Antimalarial activity of novel 4-cyano-3-methylisoquinoline inhibitors against Plasmodium falciparum: design, synthesis and biological evaluation.
Central to malaria pathogenesis is the invasion of human red blood cells by Plasmodium falciparum parasites. Following each cycle of intracellular development and replication, parasites activate a cellular program to egress from their current host cell and invade a new one. The orchestration of this process critically relies upon numerous organised phospho-signaling cascades, which are mediated by a number of central kinases. Parasite kinases are emerging as novel antimalarial targets as they have diverged sufficiently from their mammalian counterparts to allow selectable therapeutic action. Parasite protein kinase A (PfPKA) is highly expressed late in the cell cycle of the parasite blood stage and has been shown to phosphorylate a critical invasion protein, Apical Membrane Antigen 1. This enzyme could therefore be a valuable drug target so we have repurposed a substituted 4-cyano-3-methylisoquinoline that has been shown to inhibit rat PKA with the goal of targeting PfPKA. We synthesised a novel series of compounds and, although many potently inhibit the growth of chloroquine sensitive and resistant strains of P. falciparum, they were found to have minimal activity against PfPKA, indicating that they likely have another target important to parasite cytokinesis and invasion.


Thymic expression of myelin basic protein (MBP). Activation of MBP-specific T cells by thymic cells in the absence of exogenous MBP.
Previous studies have shown golli-myelin basic protein (MBP) mRNA to be expressed in the thymus of normal SJL mice, but translation of the mRNA was not assessed. To test for the presence of immunoreactive protein, single cell suspensions were prepared from adult SJL thymus and cultured with syngeneic MBP-specific T cells. After 48 h [3H]thymidine was added to the microcultures to assess T cell proliferation. MBP-specific T cell lines proliferated strongly (stimulation index range, 13-31). T cell lines specific for MBP exon 2, MBP peptide 89-101, proteolipid protein peptide 139-151, and OVA gave stimulation indices of 10-13, 5-6, 2-3, and 2-3, respectively. Stimulatory activity could be abrogated by irradiation of either the thymic cells or the MBP-specific T cells. Stimulatory activity was a property of a minor population of plastic-adherent thymic cells. Monoclonal anti-I-As Ab added to the microcultures inhibited the reaction by 77%. MBP-specific T cells cultured with syngeneic nonirradiated thymus cells in the absence of added MBP transferred experimental autoimmune encephalomyelitis adoptively to syngeneic recipients. These findings indicate that golli-MBP mRNA is translated in normal SJL thymus, and that peptides reactive with MBP-specific T cells in the context of class II MHC molecules are expressed.


[Psychoanalysis and University: the potential interface through psychoanalytic clinical research. Alice Glass-breaker].
The relationships between Psychoanalysis, Science and Universities have been discussed since Freud and gained relevance over the 21st century as an important subject for Psychoanalysis. Psychoanalysts who work in Universities are supposed to conduct research. This article comments on possible psychoanalytical research models and supports individual case clinical research through the psychoanalytical method itself, added on qualitative research elements. The authors study the psychoanalytical psychotherapy process and the mental functioning of a borderline patient treated for four years in a public university institution. Approximately 120 of 340 sessions were registered. They were read and re-read by the investigator-psychoanalyst as if they were narratives and revealed some meaningful developments from the psychoanalyst-patient.


Model for branch initiation in Aspergillus nidulans based on measurements of growth parameters.
The rate of hyphal elongation and the number of branches per hypha were measured on short sporelings of Aspergillus nidulans growing at different rates. The rate of elongation was proportional to total length in unbranched and branched hyphae. At each growth rate, the number of branches per hypha increased with increasing length and gave approximately straight-line graphs when plotted against length. The average number of branches per unit of hyphal length was quite different for the various growth rates and increased in direct proportion to the growth rate. The results are interpreted to mean that (i) growing tips have a maximum rate at which they can elongate and which is reached at hyphal lengths characteristic of the particular growth rate and (ii) a new branch is formed when the capacity of the hypha to elongate exceeds that of the existing tips.


Modulating Protein Phosphatase 2A Rescues Disease Phenotype in Neurodegenerative Tauopathies.
Alzheimer's disease (AD) is the leading cause of dementia worldwide accounting for around 70% of all cases. There is currently no treatment for AD beyond symptom management and attempts at developing disease-modifying therapies have yielded very little. These strategies have traditionally targeted the peptide Aβ, which is thought to drive pathology. However, the lack of clinical translation of these Aβ-centric strategies underscores the need for diverse treatment strategies targeting other aspects of the disease. Metal dyshomeostasis is a common feature of several neurodegenerative diseases such as AD, Parkinson's disease, and frontotemporal dementia, and manipulation of metal homeostasis has been explored as a potential therapeutic avenue for these diseases. The copper ionophore glyoxalbis-[N4-methylthiosemicarbazonato]Cu(II) (CuII(gtsm)) has previously been shown to improve the cognitive deficits seen in an AD animal model; however, the molecular mechanism remained unclear. Here we report that the treatment of two animal tauopathy models (APP/PS1 and rTg4510) with CuII(gtsm) recovers the cognitive deficits seen in both neurodegenerative models. In both models, markers of tau pathology were significantly reduced with CuII(gtsm) treatment, and in the APP/PS1 model, the levels of Aβ remained unchanged. Analysis of tau kinases (GSK3β and CDK5) revealed no drug induced changes; however, both models exhibited a significant increase in the levels of the structural subunit of the tau phosphatase, PP2A. These findings suggest that targeting the tau phosphatase PP2A has therapeutic potential for preventing memory impairments and reducing the tau pathology seen in AD and other tauopathies.


[Responsiveness and sustainability of psychiatric care in France during COVID-19 epidemic].
The sudden changes of healthcare system due to COVID-19 particularly affect the organization of psychiatry. The objective of this review is to examine the adaptations of psychiatric care in France during this pandemic. This narrative review is based on the observation of changes made in French psychiatric hospitals and on an analysis of the literature. Regarding psychiatric hospitalization, the COVID-19 epidemic required rapid measures that profoundly modified the conditions of patients' reception, forcing the medical staffs to adapt their methods of care. The authors noted the creation of at least 89 wards specifically dedicated to patients with COVID-19 needing psychiatric hospitalization, allowing dual care of general medicine and psychiatry. Regarding ambulatory care, maintaining patients with long-term follow-up was a priority. Patients recalling and teleconsultation have been precious resources but cannot entirely replace face-to-face consultations. COVID-19 epidemic created unprecedented situation of large-scale upheavals in the healthcare system and in society. Despite the absence of previous recommendations on the subject, French psychiatry has shown great adaptability. Some changes could inspire post-COVID-19 care.


Interprofessional education on a training ward for older people: students' conceptions of nurses, occupational therapists and social workers.
Collaboration between professionals in health and social care is essential to meet the needs of the patient. The collaboration is dependent on knowledge and understanding of each other's roles. One means of improving communication and collaboration among professionals is interprofessional education. The aim of this study was to describe the variation in how students in nursing, occupational therapy and social work perceived their own and the other professions. Over a three-week period two interviews were conducted with each of 16 students who were on an interprofessional training ward for older people in a municipal setting in Sweden. A phenomenographical approach was used in the analysis of the interviews. The findings showed great variation in how the students perceived the professions, from simplistic in terms of tasks to a more complex conception in terms of knowledge, responsibility and values. Differences in the ways professions were described concerning their professional stance towards the patients were especially accentuated. The findings indicate that the students need opportunities for reflection on and scrutiny of each other's beliefs and knowledge. The influence of interprofessional education involving reflection on the different health-care professions needs to be explored in future research.


Cleavage of poly(ADP-ribose) polymerase: a sensitive parameter to study cell death.
Proteases play a crucial role in apoptosis or programmed cell death. The aim of this review is to highlight the purpose for which these proteases are activated, i.e., to specifically cleave a select subset of cellular proteins at an appropriate time during cell death. Poly(ADP-ribose) polymerase (PARP), a nuclear protein implicated in DNA repair, is one of the earliest proteins targeted for a specific cleavage to the signature 89-kDa fragment during apoptosis. Characterization of the apoptotic cleavage of PARP and other target proteins helped in understanding the role of cysteine aspartic acid specific proteases (caspases) in the apoptotic process. We have recently identified that in some models of cell death, the cleavage pattern for PARP is different from production of the signature 89-kDa fragment. Necrotic death of HL-60 cells and apoptotic death of Jurkat cells mediated by granzyme B and perforin were accompanied by distinct additional fragments, suggesting cleavage of PARP at other sites by caspases or other death proteases. This review summarizes how detection and characterization of PARP cleavage could serve as a sensitive parameter for identification of different types of cell death and as a marker for activation of different death proteases. The putative biological functions for early cleavage of PARP in apoptosis are also discussed.


Effect of affinity sorbent on proteomic profiling of isatin-binding proteins of mouse brain.
Use of small molecules for isolation of particular sub-proteomes is often complicated by the need for chemical modification of a parent compound for affinity sorbent preparation. Isatin (indoledione-2,3) is an endogenous indole that exhibits a wide spectrum of biological activities. Using 5-aminocaproylisatin for proteomic profiling of fractionated rodent brain homogenates, we previously identified more than sixty individual proteins. However, proteins tested in an optical biosensor study for validation of their isatin-binding capacity demonstrated different affinity for immobilized 5-aminocaproylisatin and 5-aminoisatin. In this study, we comparatively evaluated proteomic profiles of isatin-binding proteins separated using both isatin analogs as the affinity ligands. The total number of identified proteins was higher with the shorter isatin analog (88 versus 66), and only 22 proteins were identical in the two proteomic profiles. Thus, proteomic profiling of brain isatin-binding proteins is significantly influenced by the length of the spacer between the amino group used for affinity ligand coupling to Sepharose and the isatin moiety. This suggests that the actual number of brain proteins interacting with endogenous (unmodified) isatin still remains underestimated due to different affinity of proteins for the isatin analogs used for the affinity-based proteomic profiling.


Neighborhoods, networks, and delivery methods.
We examine the contribution of information transmission among pregnant women to geographic variation in C-sections in Lombardy, Italy. Defining networks as pregnant women living in the same municipality, we observe that if the incidence of C-sections within the womans network is one standard deviation higher over the 12 months preceding delivery, then her probability of delivering by C-section is 0.007 percentage points (3%) higher. This result is mainly a network effect on Italian women, while it arises from both network and neighborhood effects on foreign women. Both groups respond to additional information, such as the incidence of C-section complications. The selection of pregnant women across hospitals does not uniquely explain our results, which are robust to alternative sample selections and specifications.


Auditory hallucinations in childhood: associations with adversity and delusional ideation.
Previous work suggests that exposure to childhood adversity is associated with the combination of delusions and hallucinations. In the present study, associations between (severity of) auditory vocal hallucinations (AVH) and (i) social adversity [traumatic experiences (TE) and stressful events (SE)] and (ii) delusional ideation were examined. A baseline case-control sample of children with and without AVH were re-assessed on AVH after 5 years and interviewed about the experience of social adversity and delusions. A total of 337 children (mean age 13.1 years, S.D.=0.5) were assessed: 40 children continued to hear voices that were present at baseline (24%, persistent group), 15 heard voices only at follow-up (9%, incident group), 130 children no longer reported AVH that were present at baseline (remitted group) and 152 never heard voices (referent group). Both TE and SE were associated with both incident and persistent AVH, as well as with greater AVH severity and delusional ideation at follow-up. In addition, the combination of AVH and delusions displayed a stronger association with TE and SE compared with either AVH or delusions alone. Early childhood AVH are mostly benign and transitory. However, experience of social adversity is associated with persistence, severity and onset of new AVH closer to puberty, and with delusional ideation.


War traumas in the Mediterranean area.
The purpose is to explore the consequences of war and its impact on mental health with attention to the Mediterranean area. Narrative review of consequences of war on mental health and on the mental health of the communities in the current crises in the Mediterranean region. A series of outbreaks of war are still raging in the Mediterranean region and producing horrible effects with a considerable number of refugees with unsatisfied needs. Studies relating to conflicts of the past suggest that the mental health consequences of these wars may affect future generations for many years. While violations of human rights are not new, what is new are attacks on medical institutions perceived to be traditionally Western. The scientific community has to fight violence through mediation of conflicts. The idea that science can improve lives is a concept that is found in the history of all Mediterranean cultures. The Greek and Roman medical tradition was saved thanks to doctors of the Arab courts when Christian fundamentalism fought science in the Middle Ages. Health institutions are the product of the great Islamic medical tradition as well as Western culture.


Measuring movement skill perceptions in preschool children: A face validity and reliability study.
The aim of this study was to determine the face validity and the reliability of a pictorial scale to assess preschool children's perceived movement skills. The pictorial scale examines the perceptions of the 12 fundamental movement skills presented in the Test of Gross Motor Development-2 (six locomotor and six object control skills) and six active play skills (e.g. cycling). A total of 91 Australian children aged four to five years from 14 preschools completed the scale on two occasions to determine test-retest reliability. Internal consistency was determined for all 18 items and for each subscale. After the second test, participants were asked open-ended questions to gain insight into their understanding of what was happening in the pictures to determine the face validity. Overall, children demonstrated a better understanding of object control and play skills compared to locomotor skills. Test-retest reliability values were good for object control (intra-class correlation, ICC = 0.75), locomotor (ICC = 0.78), active play skills (ICC = 0.80) and all 18 skills combined (ICC = 0.77). Internal consistency was acceptable for locomotor (Test 1 0.64, Test 2 0.75), object control (0.71, 0.76), active play skills (0.76, 0.74) and for all 18 skills combined (0.88, 0.90). Overall, this instrument demonstrated an acceptable reliability in preschool children and has provided preliminary information for examining the perceptions of movement skills. This age group presents a key target area for occupational therapists, who can assist preschool children to further develop fundamental movement skills, integrate these skills in their roles and occupations and increase engagement in meaningful physical activity in the long term.


Minority health and small numbers epidemiology: a case study of living conditions and the health of children in 5 foreign Romá camps in Italy.
We sought to test methods for generating epidemiological evidence on health conditions of small, dispersed minority communities. We used community-based mixed methods including a cross-sectional survey in 5 purposely selected settlements of Khorakané Romá (Gypsies of Muslim culture) in Italy to study the living conditions and health status of children aged from birth to 5 years. In the 15 days prior to the survey, 32% of the children had suffered diarrhea and 55% had had a cough. Some 17% had experienced respiratory difficulties during the past year. Risk factors associated with these outcomes included years spent living at the camp, overcrowding, housing conditions, use of wood-burning stoves, presence of rats, and issues related to quality of sanitation and drains. Qualitative information helped define the approach and the design, and in the interpretation and consolidation of quantitative results. Guided by the priorities expressed by dispersed minority communities, small studies with little resources can provide a solid base to advocate for evidence-based participatory planning. Exact intervals appeared to be robust and conservative enough compared with other intervals, conferring solidity to the results.


A green and bio-inspired process to afford durable anti-biofilm properties to stainless steel.
A bio-inspired durable anti-biofilm coating was developed for industrial stainless steel (SS) surfaces. Two polymers inspired from the adhesive and cross-linking properties of mussels were designed and assembled from aqueous solutions onto SS surfaces to afford durable coatings. Trypsin, a commercially available broad spectrum serine protease, was grafted as the final active layer of the coating. Its proteolytic activity after long immersion periods was demonstrated against several substrata, viz. a synthetic molecule, N-α-benzoyl-DL-arginine-p-nitroanilide hydrochloride (BAPNA), a protein, FTC-casein, and Gram-positive biofilm forming bacterium Staphylococcus epidermidis.


Aldosterone stimulates reactive oxygen species production through activation of NADPH oxidase in rat mesangial cells.
It has recently been shown that glomerular mesangial injury is associated with increases in renal cortical reactive oxygen species (ROS) levels in rats treated chronically with aldosterone and salt. This study was conducted to determine the mechanisms responsible for aldosterone-induced ROS production in cultured rat mesangial cells (RMC). Oxidative fluorescent dihydroethidium was used to evaluate intracellular production of superoxide anion (O(2)(-)) in intact cells. The lucigenin-derived chemiluminescence assay was used to determine NADPH oxidase activity. The staining of dihydroethidium was increased in a dose-dependent manner by aldosterone (1 to 100 nmol/L) with a peak at 3 h in RMC. Aldosterone (100 nmol/L for 3 h) also significantly increased NADPH oxidase activity from 232 +/- 18 to 346 +/- 30 cpm/5 x 10(4) cells. Immunoblotting data showed that aldosterone (100 nmol/L for 3 h) increased p47phox and p67phox protein levels in the membrane fraction by approximately 2.1- and 2.3-fold, respectively. On the other hand, mRNA expression of NADPH oxidase membrane components, p22phox, Nox-1, and Nox-4, were not altered by aldosterone (for 3 to 12 h) in RMC. Pre-incubation with the selective mineralocorticoid receptor (MR) antagonist, eplerenone (10 micromol/L), significantly attenuated aldosterone-induced O(2)(-) production, NADPH oxidase activation and membranous translocation of p47phox and p67phox. These results suggest that aldosterone-induced ROS generation is associated with NAPDH oxidase activation through MR-mediated membranous translocation of p47phox and p67phox in RMC. These cellular actions of aldosterone may play a role in the pathogenesis of glomerular mesangial injury.


Two Forms of Tyrosyl-tRNA Synthetase from Pseudomonas aeruginosa: Characterization and Discovery of Inhibitory Compounds.
Pseudomonas aeruginosa is a multidrug-resistant (MDR) pathogen and a causative agent of both nosocomial and community-acquired infections. The genes (tyrS and tyrZ) encoding both forms of P. aeruginosa tyrosyl-tRNA synthetase (TyrRS-S and TyrRS-Z) were cloned and the resulting proteins purified. TyrRS-S and TyrRS-Z were kinetically evaluated and the Km values for interaction with Tyr, ATP, and tRNATyr were 172, 204, and 1.5 μM and 29, 496, and 1.9 μM, respectively. The kcatobs values for interaction with Tyr, ATP, and tRNATyr were calculated to be 3.8, 1.0, and 0.2 s-1 and 3.1, 3.8, and 1.9 s-1, respectively. Using scintillation proximity assay (SPA) technology, a druglike 2000-compound library was screened to identify inhibitors of the enzymes. Four compounds (BCD37H06, BCD38C11, BCD49D09, and BCD54B04) were identified with inhibitory activity against TyrRS-S. BCD38C11 also inhibited TyrRS-Z. The IC50 values for BCD37H06, BCD38C11, BCD49D09, and BCD54B04 against TyrRS-S were 24, 71, 65, and 50 μM, respectively, while the IC50 value for BCD38C11 against TyrRS-Z was 241 μM. Minimum inhibitory concentrations (MICs) were determined against a panel of clinically important pathogens. All four compounds were observed to inhibit the growth of cultures of both Gram-positive and Gram-negative bacteria organisms with a bacteriostatic mode of action. When tested against human cell cultures, none of the compounds were toxic at concentrations up to 400 μg/mL. In mechanism of inhibition studies, BCD38C11 and BCD49D09 selectively inhibited TyrRS activity by competing with ATP for binding. BCD37H06 and BCD54B04 inhibited TyrRS activity by a mechanism other than substrate competition.


[Cholinesterase inhibitors for Alzheimer disease: preliminary recommendations for treatment. Dutch Society for Psychiatry, Section of Geriatric Psychiatry].
Several cholinesterase inhibitors have recently become available for Alzheimer's disease. To reach consensus about their relevance and use in daily practice, a meeting with clinical experts was organised by the section of Geriatric Psychiatry of the Dutch Society for Psychiatry. So far, available drugs have only very modest effects on cognitive functioning and clinical impression, compared with placebo. The question whether these effects are clinically relevant cannot be answered yet. Awaiting the results of further studies, it was decided not to use cholinesterase inhibitors on a routine basis but to limit prescription of these drugs to research setting or under well-controlled conditions with regard to diagnosis and evaluation. Furthermore, prescription must be limited to mild or moderately severe dementia caused by 'probable Alzheimer's disease'. Proper evaluation in the individual patient is still an unresolved problem. Therefore, n = I protocols are to be designed. These should include the use of appropriate and standardised instruments measuring cognitive functions, behavioural functions and activities of daily life. The skills and experience required will be available in specialised and multidisciplinary units for dementia. Pharmaceutical treatment for Alzheimer's disease must be integrated with all other available forms of patient care.


[The characteristics of the engorgement of the eye vessels in the dynamics of the burn process].
Peculiarities of eye blood filling in dynamics of a burn process of different severity have been studied in 29 patients with mild burns, in 19--with moderate, in 56--with severe, and in 32--with extremely severe. It is found that changes in ocular vessels blood filling correlate with severity of the burn, the degree of the development of the burn process. The usage of rheopolyglucine and cavinton in a complex treatment of eye burns positively influences the course of the burn disease. The degree of the increase of eye blood filling is in inverse dependence on the degree of destructive changes in eye tissues. Restoration of blood filling as a result of the usage of rheopolyglucin and cavinton in the shortest terms after burn stimulates the stoppage of inflammatory-destructive changes and acceleration of regenerative processes.


The effect of body mass index on survival following heart transplantation: do outcomes support consensus guidelines?
To determine the relationship between body mass index (BMI) at the time of transplant and posttransplant survival and morbidity. The recent International Society for Heart and Lung Transplantation listing criteria for heart transplantation stated that candidates should achieve a BMI <30 kg/m-or percent ideal body weight <140%-before listing for cardiac transplantation. However, data to support these recommendations are limited and often conflicting. United Network of Organ Sharing provided de-identified patient-level data. Analysis included 19,593 orthotopic heart transplant recipients aged >or=18 years and transplanted January 1 1995-December 31 2005. Follow-up data were provided through February 8, 2008. Recipients were stratified by BMI at the time of transplantation: BMI <18.5 (underweight), 18.5 to 24.99 (normal weight), 25 to 29.99 (overweight), 30 to 34.99 (obesity class I), and >or=35 (obesity class II/III). The primary outcome measure was post-transplant survival. Risk-adjusted median survival in the underweight, normal weight, overweight, obesity I, and obesity II/III groups was 8.31, 10.20, 10.03, 9.51, and 9.05 years, respectively. In multivariate Cox proportional hazards regression, BMI in the overweight (HR = 1.08, 0.99-1.17; P = 0.055) and obesity I (HR = 1.05, 0.99-1.12; P = 0.091) ranges were not associated with significantly diminished survival. However, BMI in the underweight (HR = 1.26, 1.11-1.43; P < 0.001) and obesity II/III (HR = 1.18, 1.01-1.38; P = 0.030) ranges were associated with diminished posttransplant survival. Findings from this analysis do not suggest that obesity I (BMI of 30-34.99) is associated with significantly higher morbidity and mortality. However, underweight and obesity II/III recipients have significantly higher morbidity and mortality compared with other groups.


Pneumococcal vaccination programs and the burden of invasive pneumococcal disease in Ontario, Canada, 1995-2011.
In 1995, a publicly funded pneumococcal vaccination program for 23-valent polysaccharide vaccine (PPV23) was introduced in Ontario. Conjugate vaccines were authorized in 2001 (PCV7), 2009 (PCV10) and 2010 (PCV13). From 1995-2011, active, population-based surveillance for invasive pneumococcal disease (IPD) was conducted in Metropolitan Toronto and Peel Region, Canada. 6404 IPD cases were included. After PPV23 program implementation in 1995, IPD due to PPV23 strains decreased 49% in older adults prior to PCV7 introduction. Estimated PPV23 efficacy in vaccine eligible adults was 42.2% (95% CI; 28.6-53.2%). IPD incidence due to PCV7 serotypes in children <5 years decreased significantly after PCV7 authorization and before introduction of a publicly funded PCV7 program. Seven years after PCV7 program implementation, the incidence of IPD due to PCV7 serotypes decreased to zero in children and by 88% in adults, however, overall IPD incidence remained unchanged in adults. In 2011, the incidence of IPD was 4.5 per 100,000 in adults aged 15-64 and 19.9 per 100,000 in adults aged over 65 years, with 45 serotypes causing disease. Between 1995 and 2011, the case fatality rate of IPD in adults decreased 2% per year (95% CI, -0.9% to -3.2%). In multivariable analysis, predictors of mortality included older age, chronic conditions, nursing home residence, current smoking, bacteraemia, and illness due to serotypes 3,11A, 19A, and 19F. While vaccination programs resulted in substantial public health benefits, herd immunity benefits of PCV7 were seen at low pediatric vaccination rates, and the case fatality rate of IPD has decreased, IPD will continue to be a cause of considerable morbidity and mortality in adults.


Assessment of effects of moon phases on hospital outpatient visits: An observational national study.
A vast amount of literature has been conducted for investigating the association of different lunar phases with human health; and it has mixed reviews for association and non-association of diseases with lunar phases. This study investigates the existence of any impact of moon phases on humans by exploring the difference in the rate of outpatient visits and type of diseases that prevail in either non-moon or moon phases. We retrieved dates of non-moon and moon phases for eight years (1st January 2001-31st December 2008) from the timeanddate.com website for Taiwan. The study cohort consisted of 1 million people from Taiwan's National Health Insurance Research Database (NHIRD) followed over eight years (1st January 2001-31st December 2008). We used the two-tailed, paired-t-test to compare the significance of difference among outpatient visits for 1229 moon phase days and 1074 non-moon phase days by using International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM) codes from NHIRD records. We found 58 diseases that showed statistical differences in number of outpatient visits in the non-moon and moon phases. The results of our study identified diseases that have significant variations during different lunar phases (non-moon and moon phases) for outpatient visits in the hospital. In order to fully understand the reality of the pervasive myth of lunar effects on human health, behaviors and diseases, more in-depth research investigations are required for providing comprehensive evidence covering all the factors, such as biological, psychological and environmental aspects.


Does Societal Cost Information Affect Patient Decision-Making in Carpal Tunnel Syndrome? A Randomized Controlled Trial.
Background: Despite studies demonstrating the effects of out-of-pocket costs on decision-making, the effect of societal cost information on patient decision-making is unknown. Given the considerable societal impact of cost of care for carpal tunnel syndrome (CTS), providing societal cost data to patients with CTS could affect decision-making and provide a strategy for reducing national health care costs. Therefore, we assessed the following hypotheses: (1) there is no difference in treatment choice (surgery vs no surgery) in a hypothetical case of mild CTS between patients randomized to receive societal cost information compared with those who did not receive this information; (2) there are no factors (eg, sex, experience with a previous diagnosis of CTS, or receiving societal cost information) independently associated with the choice for surgery; and (3) there is no difference in attitudes toward health care costs between patients choosing surgery and those who did not. Methods: In this randomized controlled trial using a hypothetical scenario, we prospectively enrolled 184 new and return patients with a nontraumatic upper extremity diagnosis. We recorded patient demographics, treatment choice in the hypothetical case of mild CTS, and their attitudes toward health care costs. Results: Treatment choice was not affected by receiving societal cost information. None of the demographic or illness factors assessed were independently associated with the choice for surgery. Patients declining surgery felt more strongly that doctors should consider their out-of-pocket costs when making recommendations. Conclusions: Providing societal cost information does not seem to affect decision-making and may not reduce the overall health care costs. For patients with CTS, health policy could nudge toward better resource utilization and finding the best care pathways for nonoperative and invasive treatments.


Orthogonal synthesis of a heterodimeric ligand for the development of the Gd(III)-Ga(III) ditopic complex as a potential pH-sensitive MRI/PET probe.
A heterodimeric polyaminocarboxylate ligand based on a DO3A-sulfonamide linked to AAZTA (6-amino-6-methylperhydro-1,4-diazepinetetracarboxylic acid) was synthesised via an orthogonal pathway in order to differentiate the two chelating cages and allow the formation of a Gd(III)-Ga(III) heteroditopic complex. The goal is to create a smart MRI/PET probe with pH dependent relaxivity and with the bimodal imaging approach that enables direct quantification of the stimulus, in this case pH. A (1)H NMR relaxometric study of the Gd-Ga heteroditopic complex addressed the pH modulation of the relaxivity and thus its possible use as an MRI pH sensitive probe.


Druggability assessment of precursor membrane protein as a target for inhibiting the Zika virus.
The Zika virus (ZIKV), a significant zoonotic flavivirus, was neglected as a human pathogen until the recent epidemic. The rapid geographic spread of the virus and association with neurological disorders has created a global public health concern pressing the need for anti-ZIKV drugs. Previous ZIKV drug discovery research has focused on three primary targets, RNA-dependent RNA polymerase, envelope protein, and viral proteases, and none has yet resulted in a commercially viable inhibitor. In the quest for finding effective inhibitors, it is important to expand the number of targets available for drug discovery research. To this end, the ZIKV precursor membrane protein (prM) comes to the forefront as a potential target due to its critical role in virus infectivity and pathogenicity. prM acts as a chaperone for envelope protein folding and prevents premature fusion of virions to the host membrane and has not been attempted as a drug target before. One critical requirement for a protein to be an effective target is the ability of the protein to be druggable, i.e. having active sites that can bind to specific ligands. In this work, the druggability of prM was assessed via molecular docking combined molecular dynamics simulations followed binding affinity kinetics studies. Compounds that had a high affinity to the prM protein were screened in silico and ligand-binding free energies were computed using molecular mechanics with generalized Born and surface area continuum solvation (MM-GBSA) method. In vitro binding kinetics via biolayer interferometry (BLI) and interaction analysis confirmed that prM could be targeted for drug discovery to combat ZIKV infection.Communicated by Ramaswamy H. Sarma.


Staphylococcus aureus isolates from patients with Kawasaki disease express high levels of protein A.
Kawasaki disease (KD) is an acute vasculitis of young children that can be complicated by coronary artery abnormalities. Recent findings suggest that a superantigen(s) may play an important role in stimulating the immune activation associated with the disease, although the origin of this superantigen(s) is unclear. Staphylococcus aureus, isolated from the rectum or pharynx of patients with KD, secretes toxic shock syndrome toxin 1 (TSST-1). The KD isolates express low levels of other exoproteins compared to isolates from patients with toxic shock syndrome (TSS). Thus, it was previously suggested that the KD isolates may be defective in the global regulatory locus agr (for accessory gene regulator), which positively regulates these factors (D. Y. M. Leung et al., Lancet 342:1385-1388, 1993). Here we describe another characteristic of KD isolates. When considered collectively, the KD isolates were found to express higher levels of staphylococcal protein A than the TSS isolates, another characteristic of an agr-defective phenotype. This correlated with a higher level of spa mRNA in these isolates. In contrast, the KD and TSS isolates expressed comparable levels of TSST-1, consistent with previous findings (D. Y. M. Leung et al., Lancet 342:1385-1388, 1993). Analysis of RNAIII transcript levels and nucleotide sequence analysis of the RNAIII-coding region suggested that the KD isolates are not defective in RNAIII, the effector molecule of the agr regulatory system. However, induction of RNAIII transcription in the KD isolates did not result in a dramatic decrease in the amount of spa mRNA, as has been reported for other strains (F. Vandenesch, J. Kornblum, and R. P. Novick, J. Bacteriol. 173:6313-6320, 1991).


Differential analysis of the frequency-dependent effects of class 1 antiarrhythmic drugs according to periodical ligand binding: implications for antiarrhythmic and proarrhythmic efficacy.
The frequency-dependent block of cardiac sodium channels by class 1 antiarrhythmic drugs can be described by a periodical ligand binding process between drug molecules and channel binding sites. This predicts a linear relation between onset-rate constant of frequency-dependent block and diastolic interval as well as saturation of block with high stimulation rates. From both relationships, the binding kinetics (time constant, tau on) and saturation level of block (bdinf) can be estimated. This is exemplified for the frequency-dependent block (reduction of the maximal upstroke velocity of action potentials) induced by prajmaline (10(-6) M). In the same way, the frequency-dependent effects of 11 other class 1 drugs reported in the literature were analyzed and compared with each other. When the drugs are ranked with respect to their binding kinetics (tau on), there is a close relationship to the subclasses (1a, 1b, 1c), with 1b drugs exhibiting the fastest and 1c drugs the slowest kinetics. However, differences also exist in the saturation behavior of frequency-dependent block even within the same subclasses (1a and 1c). Thus, the class 1 drugs can also be subdivided in three other groups exhibiting clearly separated bands of block-frequency relations, with half-maximal saturation occurring at different stimulation rates. Our findings may have differential implications for the antiarrhythmic and proarrhythmic efficacy of class 1 drugs.


Notch regulates numb: integration of conditional and autonomous cell fate specification.
The Notch cell-cell signaling pathway is used extensively in cell fate specification during metazoan development. In many cell lineages, the conditional role of Notch signaling is integrated with the autonomous action of the Numb protein, a Notch pathway antagonist. During Drosophila sensory bristle development, precursor cells segregate Numb asymmetrically to one of their progeny cells, rendering it unresponsive to reciprocal Notch signaling between the two daughters. This ensures that one daughter adopts a Notch-independent, and the other a Notch-dependent, cell fate. In a genome-wide survey for potential Notch pathway targets, the second intron of the numb gene was found to contain a statistically significant cluster of binding sites for Suppressor of Hairless, the transducing transcription factor for the pathway. We show that this region contains a Notch-responsive cis-regulatory module that directs numb transcription in the pIIa and pIIIb cells of the bristle lineage. These are the two precursor cells that do not inherit Numb, yet must make Numb to segregate to one daughter during their own division. Our findings reveal a new mechanism by which conditional and autonomous modes of fate specification are integrated within cell lineages.