Oxidative DNA damage in prostate cancer patients consuming tomato sauce-based entrees as a whole-food intervention.
Human prostate tissues are vulnerable to oxidative DNA damage. The risk of prostate cancer is lower in men reporting higher consumption of tomato products, which contain high levels of the antioxidant lycopene. We examined the effects of consumption of tomato sauce-based pasta dishes on lycopene uptake, oxidative DNA damage, and prostate-specific antigen (PSA) levels in patients already diagnosed with prostate cancer. Thirty-two patients with localized prostate adenocarcinoma consumed tomato sauce-based pasta dishes for the 3 weeks (30 mg of lycopene per day) preceding their scheduled radical prostatectomy. Serum and prostate lycopene concentrations, serum PSA levels, and leukocyte DNA oxidative damage (ratio of 8-hydroxy-2'-deoxyguanosine [8-OHdG] to 2'-deoxyguanosine [dG]) were assessed before and after the dietary intervention. DNA oxidative damage was assessed in resected prostate tissue from study participants and from seven randomly selected prostate cancer patients. All statistical tests were two-sided. After the dietary intervention, serum and prostate lycopene concentrations were statistically significantly increased, from 638 nM (95% confidence interval [CI] = 512 to 764 nM) to 1258 nM (95% CI = 1061 to 1455 nM) (P<.001) and from 0.28 nmol/g (95% CI = 0.18 to 0.37 nmol/g) to 0.82 nmol/g (95% CI = 0.57 to 1.11 nmol/g) (P <.001), respectively. Compared with preintervention levels, leukocyte oxidative DNA damage was statistically significantly reduced after the intervention, from 0.61 8-OHdG/10(5) dG (95% CI = 0.45 to 0.77 8-OHdG/10(5) dG) to 0.48 8-OHdG/ 10(5) dG (95% CI = 0.41 to 0.56 8-OHdG/10(5) dG) (P =.005). Furthermore, prostate tissue oxidative DNA damage was also statistically significantly lower in men who had the intervention (0.76 8-OHdG/10(5) dG [95% CI = 0.55 to 0.96 8-OHdG/10(5) dG]) than in the randomly selected patients (1.06 8-OHdG/10(5) dG [95% CI = 0.62 to 1.51 8-OHdG/10(5) dG]; P =.03). Serum PSA levels decreased after the intervention, from 10.9 ng/mL (95% CI = 8.7 to 13.2 ng/mL) to 8.7 ng/mL (95% CI = 6.8 to 10.6 ng/mL) (P<.001). These data indicate a possible role for a tomato sauce constituent, possibly lycopene, in the treatment of prostate cancer and warrant further testing with a larger sample of patients, including a control group.


Temporal whole field sawtooth flicker without a spatial component elicits a myopic shift following optical defocus irrespective of waveform direction in chicks.
Myopia (short-sightedness) is the commonest visual disorder and greatest risk factor for sight threatening secondary pathologies. Myopia and hyperopia can be induced in animal models by rearing with optical lens defocus of opposite sign. The degree of refractive compensation to lens-induced defocus in chicks has been shown to be modified by directionally drifting sawtooth spatio-temporal luminance diamond plaids, with Fast-ON sawtooth spatio-temporal luminance profiles inhibiting the myopic shift in response to negative lenses, and Fast-OFF profiles inhibiting the hyperopic shift in response to positive lenses. What is unknown is whether similar sign-of-defocus dependent results produced by spatio-temporal modulation of sawtooth patterns could be achieved by rearing chicks under whole field low temporal frequency sawtooth luminance profiles at 1 or 4 Hz without a spatial component, or whether such stimuli would indiscriminately elicit a myopic shift such as that previously shown with symmetrical (or near-symmetrical) low frequency flicker across a range of species. Hatchling chicks (n = 166) were reared from days five to nine under one of three defocus conditions (No Lens, +10D lens, or -10D lens) and five light conditions (No Flicker, 1 Hz Fast-ON/Slow-OFF sawtooth flicker, 4 Hz Fast-ON/Slow-OFF sawtooth flicker, 1 Hz Fast-OFF/Slow-ON sawtooth flicker, or 4Hz Fast-OFF/Slow-ON sawtooth flicker). The sawtooth flicker was produced by light emitting diodes (white LEDs, 1.2 -183 Lux), and had no measurable dark phase. Biometrics (refraction and ocular axial dimensions) were measured on day nine. Both 1 Hz and 4 Hz Fast-ON and Fast-OFF sawtooth flicker induced an increase in vitreous chamber depth that was greater in the presence of negative compared to positive lens defocus. Both sawtooth profiles at both temporal frequencies inhibited the hyperopic shift in response to +10D lenses, whilst full myopic compensation (or over-compensation) in response to -10D lenses was observed. Whole field low temporal frequency Fast-ON and Fast-OFF sawtooth flicker induces a generalized myopic shift, similar to that previously shown for symmetrical sine-wave and square-wave flicker. Our findings highlight that temporal modulation of retinal ON/OFF pathways per se (without a spatial component) is insufficient to produce strong sign-of-defocus dependent effect.


Critical roles of actin-interacting protein 1 in cytokinesis and chemotactic migration of mammalian cells.
Cofilin regulates actin filament dynamics by stimulating actin filament disassembly and plays a critical role in cytokinesis and chemotactic migration. Aip1 (actin-interacting protein 1), also called WDR1 (WD-repeat protein 1), is a highly conserved WD-repeat protein in eukaryotes and promotes cofilin-mediated actin filament disassembly in vitro; however, little is known about the mechanisms by which Aip1 functions in cytokinesis and cell migration in mammalian cells. In the present study, we investigated the roles of Aip1 in cytokinesis and chemotactic migration of human cells by silencing the expression of Aip1 using siRNA (small interfering RNA). Knockdown of Aip1 in HeLa cells increased the percentage of multinucleate cells; this effect was reversed by expression of an active form of cofilin. In Aip1-knockdown cells, the cleavage furrow ingressed normally from anaphase to early telophase; however, an excessive accumulation of actin filaments was observed on the contractile ring in late telophase. These results suggest that Aip1 plays a crucial role in the completion of cytokinesis by promoting cofilin-mediated actin filament disassembly in telophase. We have also shown that Aip1 knockdown significantly suppressed chemokine-induced chemotactic migration of Jurkat T-lymphoma cells, and this was blocked by expression of an active form of cofilin. Whereas control cells mostly formed a single lamellipodium in response to chemokine stimulation, Aip1 knockdown cells abnormally exhibited multiple protrusions around the cells before and after cell stimulation. This indicates that Aip1 plays an important role in directional cell migration by restricting the stimulus-induced membrane protrusion to one direction via promoting cofilin activity.


Preoperative chemoradiation therapy for esophageal cancer is a risk factor for the elevation of high mobility group box-1, leading to an increase in postoperative severe pulmonary complications.
We herein clarified the time course of changes in the serum high mobility group box chromosomal protein-1 (HMGB-1) concentrations in esophageal cancer patients after esophagectomy, and investigated whether the perioperative serum HMGB-1 levels correlate with the administration of neoadjuvant chemoradiation therapy (NACRT) and the postoperative clinical course, especially the occurrence of pulmonary complications, in such patients. Sixty patients who underwent right transthoracic esophagectomy for esophageal cancer were enrolled in this study. The relationship between the perioperative serum HMGB-1 levels and NACRT, and the postoperative severe pulmonary complications were evaluated. Patients with severe pulmonary complications (n = 44) tended to have undergone NACRT more often than those without severe pulmonary complications (n = 16). The preoperative and postoperative day 7 serum HMGB-1 concentrations were significantly higher in patients with severe pulmonary complications than those in patients without severe pulmonary complications. In the univariate and multivariate analyses, the use of NACRT and the preoperative elevations in the serum HMGB-1 levels (>4.2 ng/mL) were found to be significantly associated with pulmonary dysfunction. Furthermore, the response to NACRT was found to be significantly associated with the preoperative serum HMGB-1 levels. The use of NACRT contributes to preoperative serum HMGB-1 elevation, and these were risk factors for the occurrence of severe postoperative pulmonary complications in patients with esophageal cancer after thoracic esophagectomy.


Natriuretic peptide system in fetal heart and circulation.
Atrial natriuretic peptide, brain natriuretic peptide and C-type natriuretic peptide belong to a family of hormones that have diuretic, natriuretic and vasodepressor activity and play a part in pressure and volume homeostasis in adults. As little is known about the natriuretic peptides during cardiac maturation, this review summarizes current knowledge about the early expression of components of the natriuretic peptide system in the heart during embryonic and fetal development. The data indicate a functional importance of the fetal natriuretic peptide system, especially under pathophysiological conditions. Thus, in the fetus, the system fulfils important beneficial compensatory roles in cardiovascular disease, rather than in day-to-day pressure and volume homeostasis. In comparison with data on the relevance of natriuretic peptides in adults, those summarized here indicate a functional maturation of the natriuretic peptide system during ontogeny in mammals.


[Esophageal perforation and pneumothorax. Complications due to placement of a stomach tube].
Perforation of the oesophagus, as well as a pneumothorax are described as complications of the use of a nasogastric feeding tube. Clinical tests to determine correct placement are noted to be unreliable and the importance of radiological control, especially in the unconscious patient, ist stressed. The suspected diagnosis of oesophageal perforation should be verified without delay to immediately initiate conservative or surgical treatment.


The structural distribution of cooperative interactions in proteins: analysis of the native state ensemble.
Cooperative interactions link the behavior of different amino acid residues within a protein molecule. As a result, the effects of chemical or physical perturbations to any given residue are propagated to other residues by an intricate network of interactions. Very often, amino acids "sense" the effects of perturbations occurring at very distant locations in the protein molecule. In these studies, we have investigated by computer simulation the structural distribution of those interactions. We show here that cooperative interactions are not intrinsically bi-directional and that different residues play different roles within the intricate network of interactions existing in a protein. The effect of a perturbation to residue j on residue k is not necessarily equal to the effect of the same perturbation to residue k on residue j. In this paper, we introduce a computer algorithm aimed at mapping the network of cooperative interactions within a protein. This algorithm exhaustively performs single site thermodynamic mutations to each residue in the protein and examines the effects of those mutations on the distribution of conformational states. The algorithm has been applied to three different proteins (lambda repressor fragment 6-85, chymotrypsin inhibitor 2, and barnase). This algorithm accounts well for the observed behavior of these proteins.


Antarctic Glacial Meltwater Impacts the Diversity of Fungal Parasites Associated With Benthic Diatoms in Shallow Coastal Zones.
Aquatic ecosystems are frequently overlooked as fungal habitats, although there is increasing evidence that their diversity and ecological importance are greater than previously considered. Aquatic fungi are critical and abundant components of nutrient cycling and food web dynamics, e.g., exerting top-down control on phytoplankton communities and forming symbioses with many marine microorganisms. However, their relevance for microphytobenthic communities is almost unexplored. In the light of global warming, polar regions face extreme changes in abiotic factors with a severe impact on biodiversity and ecosystem functioning. Therefore, this study aimed to describe, for the first time, fungal diversity in Antarctic benthic habitats along the salinity gradient and to determine the co-occurrence of fungal parasites with their algal hosts, which were dominated by benthic diatoms. Our results reveal that Ascomycota and Chytridiomycota are the most abundant fungal taxa in these habitats. We show that also in Antarctic waters, salinity has a major impact on shaping not just fungal but rather the whole eukaryotic community composition, with a diversity of aquatic fungi increasing as salinity decreases. Moreover, we determined correlations between putative fungal parasites and potential benthic diatom hosts, highlighting the need for further systematic analysis of fungal diversity along with studies on taxonomy and ecological roles of Chytridiomycota.


Early detection of memory deficits and memory improvement with vaccinia virus complement control protein in an Alzheimer's disease model.
Vaccinia virus complement control protein (VCP) inhibits both the classical and alternate complement pathways. In diseases such as traumatic brain injury (TBI) and Alzheimer's disease (AD), pathological inflammation is caused by amongst several factors, prolonged or inappropriate activation of the complement system and is a significant cause of neurodegeneration. This study investigates for the first time the use of a cheeseboard maze to evaluate cognitive deficits and the effect of VCP on memory processes in 2- and 3-month-old mice that express mutant amyloid precursor protein (APPswe) and mutant presenilin 1 (Ps1dE9) that correspond to a form of early onset AD. A four-phase training schedule was carried out on the cheeseboard maze before intracranial injections of 5 microl of VCP (1.7 microg/microl) or 5 microl saline. Two weeks later the effect of VCP on memory was evaluated. A statistically significant decrease in goal latency in VCP-treated mice than saline-treated transgenic mice in both the first probe and reverse tasks was observed. Similarly, after a second intracranial VCP or saline injection performed 2 months later, the 6.5- and 7.5-month aged VCP-injected mice performed significantly better in goal latency in both second probe and reverse tasks than saline-treated mice. These data also demonstrated that the use of a dry maze is a sensitive technique for distinguishing cognitive measures between non-transgenic and APPswe/PS1De9 transgenic mice at a much earlier stage.


Does the frailty phenotype at the age of 66 to 71 predict death? A 14-year survival analysis of the Lc65+ study.
Frailty is a health characteristic resulting from the loss of physiological reserve of multiple organs, leading to exposure to adverse outcomes, and is possibly reversible in its earliest stages. It is identified by a specific phenotype that contributes to the practice of geriatric medicine, where it is considered a potential target for preventive action. This phenotype has recently attracted interest in other medical specialties for risk assessment before stressful interventions in older adults. Whereas frailty is unusual in sexagenarians, pre-frailty is common. This longitudinal study aimed to evaluate the significance of fulfilling at least one criterion of the frailty phenotype in the late sixties as a predictor of short- and long-term mortality in males and females. Data came from the first sample of the Lc65+ cohort, representative of the community-dwelling Lausanne population born between 1934 and 1939 (n = 1315). After baseline assessment of the five criteria of Fried's frailty phenotype (shrinking, exhaustion, muscular weakness, motor slowness and low physical activity) in 2005 (age 66-71 years), deaths were recorded over 14 years. We separated individuals into non-frail (fulfilling 0 criterion) and (pre-)frail (1+ criteria). The relationship between the phenotype and mortality was investigated graphically using Kaplan-Meier survival curves and quantified in Cox models. Multivariable analyses incrementally controlled age, socioeconomic and health characteristics. The prediction of fully adjusted models was evaluated using the Harrell's C index. Overall, 401 persons (30.5%) were (pre-)frail at baseline. A quarter of the 1315 participants died over 14 years (n = 336, 25.6%). The mortality rate was significantly higher in males in the (pre-)frail subgroup only. Survival curves showed a significant effect of (pre-)frailty on the risk of dying for both sexes. The effect of (pre-)frailty on mortality was stronger during the first 4 years of the follow-up. In males, it was significant both in short (0-4 years) and longer (>4-14 years) terms. In females, it was significant in the short term only. In all models, the estimated effect was stronger in males. The fully adjusted model was fairly predictive of death in the short term both in males (Harrell's C 0.79) and females (0.75). The significantly higher mortality of individuals presenting 1+ frailty criteria supports the appropriateness of a systematic assessment of the frailty phenotype at the age of 66-71 years. In both females and males, early identification of pre-frailty has the potential to limit or reverse the development of frailty and extend lifespan through adequate individual management.


Factors associated with sense of community among allied health students.
Over the last decade, there has been a substantial increase in online education in the health professions, as well as growing recognition that teamwork and collaboration are essential to success. While the impact of students' sense of community on factors such as course satisfaction and retention has been studied among college enrollees in general, there is little research exploring this concept among allied health students. To address this shortcoming, a convenience sample of students enrolled in a large northeastern school of health-related professions was surveyed to gather information on their demographics, curriculum and selected course attributes, perceived instructor teaching perspectives, and sense of community. Univariate analysis indicated that entry-level students experienced a greater sense of community than post-professional students. Multivariate analysis revealed that instructor-determined factors of encouraging discussion, encouraging expression of opinions, and specifying response times best predicted sense of community. With all other variables controlled, perceptions of community were significantly lower in online courses, among students for whom English was their second language, and in courses where instructors were perceived as focused primarily on content delivery. This study supports promoting selected course and instructor-related attributes associated with sense of community in allied health education, with a particular focus on both non-native English speakers and post-professional students. Enhancement of online courses with strategies that increase instructor presence, better engage students, and facilitate interaction also are warranted.


Ferulic acid altered IL-17A/IL-17RA interaction and protected against imiquimod-induced psoriasis-like skin injury in mice.
Ferulic acid (FA), a phenolic phytochemical, is commonly found in grains, vegetables, and fruits. Interleukin-17A (IL-17A) and IL-17 receptor A (IL-17RA) interaction is one of important therapeutic targets for psoriasis. Here we analyzed the FA effects on IL-17A/IL-17RA interaction and psoriasis-like skin injury induced by imiquimod (IMQ). IL-17A-blocking assay and docking analysis showed that FA interacted with Trp-67, Gln-94, and Glu-95 residues of IL-17A via hydrogen bonds and consequently abolished the binding of IL-17RA to IL-17A. Mice were topically given with IMQ and orally given with various amounts of FA for 14 consecutive days. FA attenuated IMQ-induced psoriasis-like skin lesions in a dose-dependent manner, and the epidermal thickness of mice treated with 100 mg/kg FA was reduced by 53.48 ± 4.44% in comparison with sham. Global analysis of differentially expressed genes showed that IMQ and FA significantly affected immune response, metabolism, and mitogen-activated protein kinase signaling pathways. Immunohistochemical staining showed that FA inhibited the infiltration and the cytokine secretion of Th17 cell, dendritic cell, and granulocyte subsets in psoriatic skin tissues. In conclusion, we newly identified that oral administration of FA protected against IMQ-induced psoriatic skin injury in mice. Moreover, its protection was associated with the interference of IL-17A/IL-17RA interaction.


Accuracy of the StatStrip versus SureStep Flexx glucose meter in neonates at risk of hypoglycemia.
The study was performed to evaluate the accuracy of the StatStrip (SS) and SureStep Flexx (SF) glucose meters compared to plasma glucose in infants at risk for neonatal hypoglycemia and to determine the effect of bilirubin and hematocrit on the results. A prospective cross-sectional study was conducted on 172 venous blood glucose samples from infants who had initial low point-of-care (POC) glucose tests measured simultaneously by SS and SF. Plasma glucose levels were compared to both POC instruments, and the effect of bilirubin and hematocrit levels on mean glucose differences were analysed. Mean (SD) plasma glucose was 2.12 (0.45) mmol/L; (range, 1.11-3.06 mmol/L). Mean (1.96SD) glucose differences of the SS versus SF were 0.21 (0.70) mmol/L and -0.04 (0.78) mmol/L, respectively. SS sensitivity was 94.7 % with an 86.1 % negative predictive value (NPV) at 2.8 mmol/L, while the SF had a 100 % sensitivity and NPV at the same cut-off level. No correlations were identified between mean glucose differences and either hematocrit or bilirubin levels in both glucose meters. Both the SS and SF glucose meters have limited use when compared to plasma glucose. Hence, they can only be employed as screening tools in at-risk neonates with an appropriate, predetermined cut-off level. Hematocrit and bilirubin levels did not affect the accuracy of both devices.


Structure and function of FusB: an elongation factor G-binding fusidic acid resistance protein active in ribosomal translocation and recycling.
Fusidic acid (FA) is a bacteriostatic antibiotic that locks elongation factor G (EF-G) to the ribosome after GTP hydrolysis during elongation and ribosome recycling. The plasmid pUB101-encoded protein FusB causes FA resistance in clinical isolates of Staphylococcus aureus through an interaction with EF-G. Here, we report 1.6 and 2.3 Å crystal structures of FusB. We show that FusB is a two-domain protein lacking homology to known structures, where the N-terminal domain is a four-helix bundle and the C-terminal domain has an alpha/beta fold containing a C4 treble clef zinc finger motif and two loop regions with conserved basic residues. Using hybrid constructs between S. aureus EF-G that binds to FusB and Escherichia coli EF-G that does not, we show that the sequence determinants for FusB recognition reside in domain IV and involve the C-terminal helix of S. aureus EF-G. Further, using kinetic assays in a reconstituted translation system, we demonstrate that FusB can rescue FA inhibition of tRNA translocation as well as ribosome recycling. We propose that FusB rescues S. aureus from FA inhibition by preventing formation or facilitating dissociation of the FA-locked EF-G-ribosome complex.


The effects of a customized over-the-counter mouth guard on neuromuscular force and power production in trained men and women.
Although mouth guards were originally designed for injury prevention, even elite athletes are now using performance mouth guards to improve athletic success. Both expensive custom models and over-the-counter models are available, but the efficacy is not well known. Some athletes remain wary of the perceived potential for detriments using a mouth guard to their performance. Thus, the purpose of this study was to examine various physical performance tests when using a mouth guard including a customized over-the-counter mouth guard. Twenty-six trained men (25 ± 4 years; 1.78 ± 0.07 m; 83.3 ± 11.4 kg) and 24 trained women (23 ± 3 years; 1.65 ± 0.08 m; 62.6 ± 7.8 kg) volunteered for the investigation. The subjects completed a familiarization period and then balanced and randomized treatment conditions that included: (a) a customized Power Balance performance mouth guard (PB MG); (b) a regular over the counter boil-and-bite mouth guard (Reg MG); and (c) a no mouth guard (No MG) treatment condition. At each visit, the subjects completed a testing protocol that was sequenced in the following order: sit-and-reach flexibility, medial-lateral balance, visual reaction time, vertical jump, 10-m sprint, bench throw, and plyo press power quotient (3PQ). Heart rate and rating of perceived exertion (RPE) were recorded around the 3PQ. Significance was set at p ≤ 0.05. Expected significant sex differences existed for all power, strength, and speed variables. Bench throw power (watts) and force (newtons) were significantly higher under PB MG than either Reg MG or No MG or in both men and women. The 3PQ power and force production were higher than that for the other 2 treatments for the PB MG for men only. There were no significant differences for treatment conditions in the heart rate or RPE after the 3PQ test. Men were better able to maintain significantly higher 3PQ power production under PB MG treatment condition compared with the other 2 treatment conditions. Rate of power development was significantly higher in men for the vertical jump when using the PB MG compared with that for other treatment conditions in men only. No differences were observed in flexibility, balance, visual reaction time, or sprint time. The PB MG performance mouth guard improves performance of upper-body loaded power exercises in both men and women and lower body power exercise in men without compromising performance on any other performance parameters.


Exercise-induced leg pain in athletes: diagnostic, assessment, and management strategies.
The purpose of this review is to describe and critically evaluate current knowledge regarding diagnosis, assessment, and management of chronic overload leg injuries which are often non-specific and misleadingly referred to as 'shin splints'. We aimed to review clinical entities that come under the umbrella term 'Exercise-induced leg pain' (EILP) based on current literature and systematically searched the literature. Specifically, systematic reviews were included. Our analyses demonstrated that current knowledge on EILP is based on a low level of evidence. EILP has to be subdivided into those with pain from bone stress injuries, pain of osteo-fascial origin, pain of muscular origin, pain due to nerve compression and pain due to a temporary vascular compromise. The history is most important. Questions include the onset of symptoms, whether worse with activity, at rest or at night? What exacerbates it and what relieves it? Is the sleep disturbed? Investigations merely confirm the clinical diagnosis and/or differential diagnosis; they should not be solely relied upon. The mainstay of diagnosing bone stress injury is MRI scan. Treatment is based on unloading strategies. A standard for confirming chronic exertional compartment syndrome (CECS) is the dynamic intra-compartmental pressure study performed with specific exercises that provoke the symptoms. Surgery provides the best outcome. Medial tibial stress syndrome (MTSS) presents a challenge in both diagnosis and treatment especially where there is a substantial overlap of symptoms with deep posterior CECS. Conservative therapy should initially aim to correct functional, gait, and biomechanical overload factors. Surgery should be considered in recalcitrant cases. MRI and MR angiography are the primary investigative tools for functional popliteal artery entrapment syndrome and when confirmed, surgery provides the most satisfactory outcome. Nerve compression is induced by various factors, e.g., localized fascial entrapment, unstable proximal tibiofibular joint (intrinsic) or secondary by external compromise of the nerve, e.g., tight hosiery (extrinsic). Conservative is the treatment of choice. The localized fasciotomy is reserved for recalcitrant cases.


Attention and working memory in resident anaesthetists after night duty: group and individual effects.
To investigate the effects of a single period of night duty on measures of attention and working memory in a group of residents (registrars) in anaesthesiology. Emphasis was placed on individual deficits using a reference point of the equivalent effect of a blood alcohol concentration (BAC) >0.05% determined by other researchers. There were 33 subjects aged 26-42 years. Night duty was performed on a weekly basis. Baseline assessments were conducted at either 08 15 or 08 55 preceding night duty and repeated 24-25 hours later, just after the completion of duty. Questionnaires included items regarding duration of sleep and the Stanford Sleepiness Scale. A battery of four reaction time (RT) tasks of increasing difficulty, lasting approximately 35 minutes, was administered on a personal computer. These ranged from simple RT to progressively more complex RT tasks incorporating working memory. A significant change was regarded as >15% deterioration in respect of speed or accuracy. The mean duration of sleep preceding night duty was 7.04 hours and 1.66 hours during the period of night duty. Intergroup comparisons revealed significant prolongation in mean response speed in the first three tests. Mean accuracy was significantly reduced only in respect of the two more complex tests. A >15% deterioration in response speed occurred in up to 30% of subjects on a single task, rising to 52% (17/33) overall. Deterioration occurred in a patchy distribution in most subjects, involving no more than one or two of the four tasks. As regards accuracy, the prevalence of deterioration increased with task complexity. Results are in general agreement with previous group analyses. A new dimension was added by the analysis of a broad spectrum of individual response to sleep deprivation. The effects of sleep loss in residents cannot be overlooked, even in a relatively benign work schedule.


A moonlighting role for enzymes of glycolysis in the co-localization of mitochondria and chloroplasts.
Glycolysis is one of the primordial pathways of metabolism, playing a pivotal role in energy metabolism and biosynthesis. Glycolytic enzymes are known to form transient multi-enzyme assemblies. Here we examine the wider protein-protein interactions of plant glycolytic enzymes and reveal a moonlighting role for specific glycolytic enzymes in mediating the co-localization of mitochondria and chloroplasts. Knockout mutation of phosphoglycerate mutase or enolase resulted in a significantly reduced association of the two organelles. We provide evidence that phosphoglycerate mutase and enolase form a substrate-channelling metabolon which is part of a larger complex of proteins including pyruvate kinase. These results alongside a range of genetic complementation experiments are discussed in the context of our current understanding of chloroplast-mitochondrial interactions within photosynthetic eukaryotes.


Nationwide survey of pediatric residency training in newborn medicine: preparation for primary care practice.
Pediatric residency training programs need regular revision, but there is little evidence to support changes. The recent reduction in time spent in the newborn nursery may negatively affect the care of infants in pediatric practice. This survey was undertaken to assess the preparation of recent graduates of training programs with respect to newborn care in their current practice. A nationwide survey was sent to 1997 pediatric residency graduates in May 1999. The American Medical Association Masterfile was used to identify pediatricians who met the criteria. The survey included questions about preparation and confidence in topics obtained from the Residency Review Committee guidelines for the newborn nursery. There were questions about duration of nursery experience, learning barriers, teaching preferences, current nursery responsibility, and practice location along with other information about the respondents. After 2 mailings, 801 surveys were received. Practice distribution was comparable to those who took the pediatric boards for the first time in 1997, and response was nationwide. Overall, 45% attend deliveries and 71% care for sick newborns. Respondents with only 1 month of normal newborn nursery felt this was insufficient preparation. Bedside teaching by faculty members was rated most highly. The most commonly rated areas of poor preparation were prenatal visits, perinatal infections, and management of breastfeeding issues. The current structure of nursery training in many residency programs may be insufficient preparation for primary care practice. Results of this and future surveys of recent graduates provide evidence to direct changes in residency training.


Preparing Fock states in the micromaser.
In this paper we give a survey of our experiments performed with the micromaser on the generation of Fock states. Three methods can be used for this purpose: the trapping states leading to Fock states in a continuous wave operation; state reduction of a pulsed pumping beam and finally using a pulsed pumping beam to produce Fock states on demand where trapping states stabilize the photon number. The latter method is discussed in detail by means of Monte Carlo simulations of the maser system. The results of the simulations are presented in a video.


Chronic vagal stimulation for the treatment of low ejection fraction heart failure: results of the NEural Cardiac TherApy foR Heart Failure (NECTAR-HF) randomized controlled trial.
The neural cardiac therapy for heart failure (NECTAR-HF) was a randomized sham-controlled trial designed to evaluate whether a single dose of vagal nerve stimulation (VNS) would attenuate cardiac remodelling, improve cardiac function and increase exercise capacity in symptomatic heart failure patients with severe left ventricular (LV) systolic dysfunction despite guideline recommended medical therapy. Patients were randomized in a 2 : 1 ratio to receive therapy (VNS ON) or control (VNS OFF) for a 6-month period. The primary endpoint was the change in LV end systolic diameter (LVESD) at 6 months for control vs. therapy, with secondary endpoints of other echocardiography measurements, exercise capacity, quality-of-life assessments, 24-h Holter, and circulating biomarkers. Of the 96 implanted patients, 87 had paired datasets for the primary endpoint. Change in LVESD from baseline to 6 months was -0.04 ± 0.25 cm in the therapy group compared with -0.08 ± 0.32 cm in the control group (P = 0.60). Additional echocardiographic parameters of LV end diastolic dimension, LV end systolic volume, left ventricular end diastolic volume, LV ejection fraction, peak V02, and N-terminal pro-hormone brain natriuretic peptide failed to show superiority compared to the control group. However, there were statistically significant improvements in quality of life for the Minnesota Living with Heart Failure Questionnaire (P = 0.049), New York Heart Association class (P = 0.032), and the SF-36 Physical Component (P = 0.016) in the therapy group. Vagal nerve stimulation as delivered in the NECTAR-HF trial failed to demonstrate a significant effect on primary and secondary endpoint measures of cardiac remodelling and functional capacity in symptomatic heart failure patients, but quality-of-life measures showed significant improvement.


[Visual behavior of the premature infant].
Ocular reflexes were studied at birth in neonates of varying gestational ages and then followed every two weeks. It is possible to distinguish reflexes that may be present at birth that are dependent on gestational age and those acquired later. Each reflex corresponds to the establishment of neuronal corrections and thus it is possible to follow the development of neural structures in utero and after birth. Ocular examination of the newborn completes the examination of the central nervous system and a scale of visual age is proposed. The repeated examination of premature infants showed the development of visual function was accelerated.


Interaction Between Frailty and Access Site in Older Adults Undergoing Transcatheter Aortic Valve Replacement.
The authors sought to determine whether frail older adults undergoing nonfemoral transcatheter aortic valve replacement (TAVR) procedures had a higher risk of 30-day and 12-month mortality. Frailty can help predict outcomes and guide therapy in older adults being considered for TAVR. Nonfemoral TAVR procedures are more invasive and impart a greater risk of adverse events, which may be less well tolerated in frail patients, compared with transfemoral TAVR procedures. This study was a post hoc analysis of the FRAILTY-AVR (Frailty Assessment Before Cardiac Surgery & Transcatheter Interventions) prospective multicenter cohort that consisted of older adults undergoing TAVR from 2012 to 2017. Frailty was assessed using the Essential Frailty Toolset (EFT). Endpoints of interest were 30-day and 12-month all-cause mortality. Interaction tables and multivariable logistic regression models were used to investigate statistical interaction on the additive and multiplicative scales. The cohort consisted of 723 patients with a mean age of 84 ± 6 years, of which 556 (77%) had femoral access and 167 (23%) had nonfemoral access. In frail patients with EFT scores ≥3 (35%), nonfemoral access was associated with increased 30-day mortality (odds ratio [OR]: 3.91; 95% confidence interval [CI]: 1.48 to 10.31); whereas in nonfrail patients with EFT scores <3 (65%), nonfemoral access had no effect (OR: 1.29; 95% CI: 0.34 to 4.94). There was statistical evidence of interaction between frailty and access site on 30-day mortality on the additive scale (relative excess risk due to interaction = 5.95). Nonfemoral access was associated with increased 1-year mortality in frail patients (OR: 1.98; 95% CI: 1.00 to 3.93) but not in nonfrail patients (OR: 1.83; 95% CI: 0.90 to 3.74), although there was no statistical evidence of interaction. Frail patients undergoing TAVR via a more invasive nonfemoral access face a substantially higher risk of 30-day mortality, whereas nonfrail older adults tolerate the procedure with a low short-term risk irrespective of access route.


Lateral cavity photonic crystal surface emitting laser based on commercial epitaxial wafer.
A lateral cavity photonic crystal surface emitting laser (LC-PCSEL) with airholes of cone-like shape etched near to the active layer is fabricated. It employs only a simple commercial epitaxial wafer without DBR and needs no wafer bonding technique. Surface emitting lasing action at 1575 nm with power of 1.8 mW is observed at room temperature, providing potential values for mass production of electrically driven PCSELs with low cost. Additionally, Fano resonance is utilized to analyze aperture equivalence of PC, and energy distribution in simplified laser structure is simulated to show oscillation and transmission characteristics of laser.


Controversies on the role of bone marrow transplantation for patients with hematopoietic malignancies.
Over the past decade, bone marrow transplantation has evolved as a treatment modality for patients with hematopoietic malignancies and for patients with responsive solid tumors. In this paper, I review (a) the indications for marrow transplantation, with emphasis on patients with hematopoietic malignancies, including acute and chronic leukemias and lymphomas; (b) the role of the donor, including the rapid expansion of compatible volunteer donors; (c) the role and design of preparative regimens; (d) the procurement of bone marrow, including marrow purging and T-cell depletion, and (e) the care of the transplant patient, including a discussion of the more commonly occurring complications.


Acute phase response to surgery of varying intensity in horses: a preliminary study.
To evaluate the postoperative inflammatory response of horses to elective surgery of varying intensity. Prospective longitudinal study. Horses referred to 2 hospitals for either arthroscopic removal of a unilateral osteochondritic lesion in the tibiotarsal joint (minimal surgical trauma, n=11), correction of recurrent laryngeal neuropathy by laryngoplasty and ventriculectomy (intermediate surgical trauma, n=10) or removal of an ovarian tumor by laparotomy (major surgical trauma, n=5). Horses had a thorough clinical examination every day. White blood cell (WBC) counts and concentrations of serum amyloid A (SAA), fibrinogen, and iron were assessed in blood samples obtained before, and 1-3, 5, 7, 9, and 11 days after surgery. Differences in levels of the inflammatory markers between the 3 surgical groups were analyzed using repeated measures ANOVA. Postoperative concentrations of SAA and fibrinogen were significantly higher in horses that had laparotomy and ovariectomy than in horses that had laryngoplasty and ventriculectomy, or arthroscopy. Iron concentrations decreased to lower levels after intermediate and major surgical trauma than after small surgical trauma. WBC count did not differ between the 3 groups. Levels of SAA, fibrinogen, and iron reflected the intensity of the surgical trauma, whereas WBC count did not. Postoperative measurements of SAA, fibrinogen, and iron may be useful for comparing surgical trauma associated with new and established surgical techniques. Moreover, knowledge of the normal postoperative acute phase response is essential, if acute phase reactants are to be used for monitoring occurrence of postoperative infections.


A Case Study Application of the Aggregate Exposure Pathway (AEP) and Adverse Outcome Pathway (AOP) Frameworks to Facilitate the Integration of Human Health and Ecological End Points for Cumulative Risk Assessment (CRA).
Cumulative risk assessment (CRA) methods promote the use of a conceptual site model (CSM) to apportion exposures and integrate risk from multiple stressors. While CSMs may encompass multiple species, evaluating end points across taxa can be challenging due to data availability and physiological differences among organisms. Adverse outcome pathways (AOPs) describe biological mechanisms leading to adverse outcomes (AOs) by assembling causal pathways with measurable intermediate steps termed key events (KEs), thereby providing a framework for integrating data across species. In this work, we used a case study focused on the perchlorate anion (ClO4-) to highlight the value of the AOP framework for cross-species data integration. Computational models and dose-response data were used to evaluate the effects of ClO4- in 12 species and revealed a dose-response concordance across KEs and taxa. The aggregate exposure pathway (AEP) tracks stressors from sources to the exposures and serves as a complement to the AOP. We discuss how the combined AEP-AOP construct helps to maximize the use of existing data and advances CRA by (1) organizing toxicity and exposure data, (2) providing a mechanistic framework of KEs for integrating data across human health and ecological end points, (3) facilitating cross-species dose-response evaluation, and (4) highlighting data gaps and technical limitations.


Genetic Variant in Human PAR (Protease-Activated Receptor) 4 Enhances Thrombus Formation Resulting in Resistance to Antiplatelet Therapeutics.
Platelet activation after stimulation of PAR (protease-activated receptor) 4 is heightened in platelets from blacks compared with those from whites. The difference in PAR4 signaling by race is partially explained by a single-nucleotide variant in PAR4 encoding for either an alanine or threonine at amino acid 120 in the second transmembrane domain. The current study sought to determine whether the difference in PAR4 signaling by this PAR4 variant is because of biased Gq signaling and whether the difference in PAR4 activity results in resistance to traditional antiplatelet intervention. Membranes expressing human PAR4-120 variants were reconstituted with either Gq or G13 to determine the kinetics of G protein activation. The kinetics of Gq and G13 activation were both increased in membranes expressing PAR4-Thr120 compared with those expressing PAR4-Ala120. Further, inhibiting PAR4-mediated platelet activation by targeting COX (cyclooxygenase) and P2Y12 receptor was less effective in platelets from subjects expressing PAR4-Thr120 compared with PAR4-Ala120. Additionally, ex vivo thrombus formation in whole blood was evaluated at high shear to determine the relationship between PAR4 variant expression and response to antiplatelet drugs. Ex vivo thrombus formation was enhanced in blood from subjects expressing PAR4-Thr120 in the presence or absence of antiplatelet therapy. Together, these data support that the signaling difference by the PAR4-120 variant results in the enhancement of both Gq and G13 activation and an increase in thrombus formation resulting in a potential resistance to traditional antiplatelet therapies targeting COX-1 and the P2Y12 receptor.


[A staff development model in psychiatric nursing].
The nursing service manager is accountable for the quality of nursing care delivered in the nursing service. It is therefore important that the nursing service manager facilitates staff development in the nursing service. It is not only the nursing service manager's responsibility to make provision for staff development--the nurse also has a responsibility in this regard. He/she should purposefully make an effort to keep up to date with the latest developments. This article focuses on the co-responsibility of the psychiatric nurse and nursing service manager regarding staff development. A model for staff development is described, in accordance with the guidelines of Dickoff, James & Wiedenbach for theory development. An inductive approach was primarily followed to describe the provisional model, after which a literature study was employed to refine and purify the model. This model was exposed to expert evaluation, after which the final model for staff development of psychiatric nurses was described. Recommendations include the testing of certain hypotheses and utilisation of this model in psychiatric nursing practice.


Cu/Zn- and Mn-superoxide dismutase distribution and concentration in adrenal tumors.
The tissue distribution of Cu/Zn- and Mn-superoxide dismutases (SOD) in adrenal tumors was studied by an immunohistochemical technique, and the concentrations of both SODs were measured by a sensitive sandwich enzyme immunoassay technique. In the normal adrenal gland, both Cu/Zn- and Mn-SODs were localized predominantly in the reticular zone of the cortex. Cu/Zn-SOD was stained clearly in the inner fascicular zone of the cortex, but not in the medulla, whereas Mn-SOD was stained weakly in the medulla. In different adrenal tumors, the localization of both stained SODs reflected the origin of the tumor cell. Thus, in one section of a pheochromocytoma only Mn-SOD was stained clearly. The concentrations of both SODs in the tissues of medullary tumors were lower than those in the normal adrenal gland and adrenocortical adenomas. The concentration of Cu/Zn-SOD in the tumor tissue of Cushing's syndrome adenoma was higher, and that of Mn-SOD was lower than the concentrations in the normal adrenal gland. The ratio of the tissue concentrations of Mn-SOD to Cu/Zn-SOD was lower in adrenal medullary tumors and Cushing's syndrome adenomas than in the normal adrenal gland and primary aldosteronism adenomas, indicating the predominance of Cu/Zn-SOD in the former, and Mn-SOD in the latter. These data suggest that the localization of Cu/Zn- and Mn-SODs in adrenal tissues reflects the specificity of the adrenal cells that produce the tissue-specific hormones. An investigation of changes in these enzymes in adrenal tumors may also provide useful information on adrenal tumor cell differentiation.


Free nipple graft reduction mammoplasty.
Free nipple graft reduction mammoplasty is the procedure of choice in patients with massive breast hypertrophy, or those high-risk patients less able to undergo a more extensive procedure. A major criticism of the technique is that it creates a flat, boxy breast that lacks projection. A technical modification of free nipple graft reduction mammoplasty is presented. In this modification, a central, bulky, superiorly based dermal-parenchymal flap extending from the "key-hole" site to the superior aspect of the areola is designed. After tailoring, this central flap is folded superiorly and secured beneath the medial and lateral flaps to create the bulk of the breast mound. This central flap can be accurately tailored to achieve the desired breast size and projection. The medial and lateral breast flaps do not create the breast mound, and are only contoured over the top of this central flap to complement the final breast form.


mTOR-dependent abnormalities in autophagy characterize human malformations of cortical development: evidence from focal cortical dysplasia and tuberous sclerosis.
Focal cortical dysplasia (FCD) is a localized malformation of cortical development and is the commonest cause of severe childhood epilepsy in surgical practice. Children with FCD are severely disabled by their epilepsy, presenting with frequent seizures early in life. The commonest form of FCD in children is characterized by the presence of an abnormal population of cells, known as balloon cells. Similar pathological changes are seen in the cortical malformations that characterize patients with tuberous sclerosis complex (TSC). However, the cellular and molecular mechanisms that underlie the malformations of FCD and TSC are not well understood. We provide evidence for a defect in autophagy in FCD and TSC. We have found that balloon cells contain vacuoles that include components of the autophagy pathway. Specifically, we show that balloon cells contain prominent lysosomes by electron microscopy, immunohistochemistry for LAMP1 and LAMP2, LysoTracker labelling and enzyme histochemistry for acid phosphatase. Furthermore, we found that balloon cells contain components of the ATG pathway and that there is cytoplasmic accumulation of the regulator of autophagy, DOR. Most importantly we found that there is abnormal accumulation of the autophagy cargo protein, p62. We show that this defect in autophagy can be, in part, reversed in vitro by inhibition of the mammalian target of rapamycin (mTOR) suggesting that abnormal activation of mTOR may contribute directly to a defect in autophagy in FCD and TSC.


Expression of gp91phox and p22phox, catalytic subunits of NADPH oxidase, on microglia in Nasu-Hakola disease brains.
The superoxide-producing nicotinamide adenine dinucleotide phosphate (NADPH) oxidase complex of phagocytes (phox) plays a key role in production of reactive oxygen species (ROS) by microglia. The catalytic subunits of the NADPH oxidase are composed of p22phox and gp91phox. Nasu-Hakola disease (NHD) is a rare autosomal recessive disorder caused by a loss-of-function mutation of either TYROBP (DAP12) or TREM2. Pathologically, the brains of NHD patients exhibit extensive demyelination designated leukoencephalopathy, astrogliosis, accumulation of axonal spheroids, and remarkable activation of microglia predominantly in the white matter of frontal and temporal lobes. However, a pathological role of the gp91phox-p22phox complex in generation of leukoencephalopathy in NHD remains unknown. We clarified the expression of gp91phox and p22phox in the white matter of the frontal cortex derived from five NHD and eight control subjects. We identified the expression of p22phox and gp91phox immunoreactivity almost exclusively on microglia. Microglia overexpressed gp91phox in NHD brains and p22phox in myotonic dystrophy (MD) brains, when compared with non-neurological control (NC) brains. These results suggest that the enhanced expression of gp91phox by microglia might contribute to overproduction of ROS highly toxic to myelinating oligodendrocytes, resulting in oligodendrocyte cell death that induces leukoencephalopathy in NHD brains.


Salicylate Sodium Suppresses Monocyte Chemoattractant Protein-1 Production by Directly Inhibiting Phosphodiesterase 3B in TNF-α-Stimulated Adipocytes.
As a worldwide health issue, obesity is associated with the infiltration of monocytes/macrophages into the adipose tissue causing unresolved inflammation. Monocyte chemoattractant protein-1 (MCP-1) exerts a crucial effect on obesity-related monocytes/macrophages infiltration. Clinically, aspirin and salsalate are beneficial for the treatment of metabolic diseases in which adipose tissue inflammation plays an essential role. Herein, we investigated the effect and precise mechanism of their active metabolite salicylate on TNF-α-elevated MCP-1 in adipocytes. The results indicated that salicylate sodium (SAS) could lower the level of MCP-1 in TNF-α-stimulated adipocytes, which resulted from a previously unrecognized target phosphodiesterase (PDE), 3B (PDE3B), rather than its known targets IKKβ and AMPK. The SAS directly bound to the PDE3B to inactivate it, thus elevating the intracellular cAMP level and activating PKA. Subsequently, the expression of MKP-1 was increased, which led to the decrease in p-EKR and p-p38. Both PDE3B silencing and the pharmacological inhibition of cAMP/PKA compromised the suppressive effect of SAS on MCP-1. In addition to PDE3B, the PDE3A and PDE4B activity was also inhibited by SAS. Our findings identify a previously unrecognized pathway through which SAS is capable of attenuating the inflammation of adipocytes.


Who Really Benefits from 3D-Based Planning of Brachytherapy for Cervical Cancer?
Although intracavitary radiotherapy (ICR) is essential for the radiation therapy of cervical cancer, few institutions in Korea perform 3-dimensional (3D)-based ICR. To identify patients who would benefit from 3D-based ICR, dosimetric parameters for tumor targets and organs at risk (OARs) were compared between 2-dimensional (2D)- and 3D-based ICR. Twenty patients with locally advanced cervical cancer who underwent external beam radiation therapy (EBRT) following 3D-based ICR were retrospectively evaluated. New 2D-based plans based on the Manchester system were developed. Tumor size was measured by magnetic resonance imaging. The mean high risk clinical target volume (HR-CTV) D90 value was about 10% lower for 2D- than for 3D-based plans (88.4% vs. 97.7%; P = 0.068). Tumor coverage did not differ between 2D- and 3D-based plans in patients with tumors ≤ 4 cm at the time of brachytherapy, but the mean HR-CTV D90 values in patients with tumors > 4 cm were significantly higher for 3D-based plans than for 2D-based plans (96.0% vs. 78.1%; P = 0.017). Similar results were found for patients with tumors > 5 cm initially. Other dosimetric parameters for OARs were similar between 2D- and 3D-based plans, except that mean sigmoid D2cc was higher for 2D- than for 3D-based plans (67.5% vs. 58.8%; P = 0.043). These findings indicate that 3D-based ICR plans improve tumor coverage while satisfying the dose constraints for OARs. 3D-based ICR should be considered in patients with tumors > 4 cm size at the time of brachytherapy or > 5 cm initially.


Evidence-based management: from theory to practice in health care.
The rise of evidence-based clinical practice in health care has caused some people to start questioning how health care managers and policymakers make decisions, and what role evidence plays in the process. Though managers and policymakers have been quick to encourage clinicians to adopt an evidence-based approach, they have been slower to apply the same ideas to their own practice. Yet, there is evidence that the same problems (of the underuse of effective interventions and the overuse of ineffective ones) are as widespread in health care management as they are in clinical practice. Because there are important differences between the culture, research base, and decision-making processes of clinicians and managers, the ideas of evidence-based practice, while relevant, need to be translated for management rather than simply transferred. The experience of the Center for Health Management Research (CHMR) is used to explore how to bring managers and researchers together and promote the use of evidence in managerial decision-making. However, health care funders, health care organizations, research funders, and academic centers need wider and more concerted action to promote the development of evidence-based managerial practice.


Beyond the H&E: Advanced Technologies for in situ Tissue Biomarker Imaging.
For decades, histopathology with routine hematoxylin and eosin staining has been and remains the gold standard for reaching a morphologic diagnosis in tissue samples from humans and veterinary species. However, within the past decade, there has been exponential growth in advanced techniques for in situ tissue biomarker imaging that bridge the divide between anatomic and molecular pathology. It is now possible to simultaneously observe localization and expression magnitude of multiple protein, nucleic acid, and molecular targets in tissue sections and apply machine learning to synthesize vast, image-derived datasets. As these technologies become more sophisticated and widely available, a team-science approach involving subspecialists with medical, engineering, and physics backgrounds is critical to upholding quality and validity in studies generating these data. The purpose of this manuscript is to detail the scientific premise, tools and training, quality control, and data collection and analysis considerations needed for the most prominent advanced imaging technologies currently applied in tissue sections: immunofluorescence, in situ hybridization, laser capture microdissection, matrix-assisted laser desorption ionization imaging mass spectrometry, and spectroscopic/optical methods. We conclude with a brief overview of future directions for ex vivo and in vivo imaging techniques.


Hypoxia-induced enhancement of nonspecific bronchial reactivity: role of leukotrienes.
Because alveolar hypoxia has been shown to cause an increase of leukotrienes in lung lavage fluid, we tested the hypothesis that enhancement of nonspecific bronchial reactivity during alveolar hypoxia may be mediated by leukotrienes. In nine conscious sheep we determined specific lung resistance (sRL) before and after exposure to either air or a hypoxic gas mixture (13% O2) for 30 min. The sheep then inhaled 50 breaths of aerosolized 5% histamine solution (n = 6) or 10 breaths of 2.5% carbachol solution (n = 6) on different days, and the measurements of sRL were repeated. On subsequent days the above protocols were repeated after pretreatment with aerosolized FPL 57231 (3 ml, 1% solution), a leukotriene receptor antagonist. Inhalation of histamine and carbachol after exposure to air caused an increase in mean sRL to 337 and 342% of base line, respectively (P less than 0.05). Exposure to the hypoxic gas mixture had no effect on sRL but enhanced the histamine- and carbachol-induced increases in mean sRL to 621 and 646% of base line, respectively (P less than 0.05); these increases were significantly higher than those observed after air exposure (P less than 0.05). FPL 57231 prevented the hypoxia-induced enhancement of bronchial reactivity to histamine and carbachol without affecting the airway responsiveness to these agents after air. In another group of eight sheep, aerosolized leukotriene C4, at a dose (50 micrograms) that per se had no affect on sRL, enhanced the bronchial reactivity to carbachol. These data suggest that in sheep during alveolar hypoxia airway hyperresponsiveness may be due to the priming of airway smooth muscle by leukotrienes.


A randomized controlled trial of light versus deep propofol sedation for elective outpatient colonoscopy: recall, procedural conditions, and recovery.
This study aimed to determine if the incidence of recall was equivalent between light and deep sedation for colonoscopy. Secondary analysis included complications, patient clinical recovery, and post-procedure cognitive impairment. Two hundred patients undergoing elective outpatient colonoscopy were randomized to light (bispectral index [BIS] 70-80) or deep (BIS < 60) sedation with propofol and fentanyl. Recall was assessed by the modified Brice questionnaire, and cognition at baseline and discharge was assessed using a Cogstate test battery. The median (interquartile range [IQR]) BIS values were different in the two groups (69 [65-74] light sedation vs 53 [46-59] deep sedation; P < 0.0001). The incidence of recall was 12% in the light sedation group and 1% in the deep sedation group. The risk difference for recall was 0.11 (90% confidence interval, 0.05 to 0.17) in the intention-to-treat analysis, thus refuting equivalence in recall between light and deep sedation (0.05 significance level; 10% equivalence margin). Overall sedation-related complications were more frequent with deep sedation than with light sedation (66% vs 47%, respectively; P = 0.008). Recovery was more rapid with light sedation than with deep sedation as determined by the mean (SD) time to reach a score of 5 on the Modified Observer's Assessment of Alertness/Sedation Scale [3 (4) min vs 7 (4) min, respectively; P < 0.001] and by the median [IQR] time to readiness for hospital discharge (65 [57-80] min vs 74 [63-86] min, respectively; P = 0.001). The incidence of post-procedural cognitive impairment was similar in those randomized to light (19%) vs deep (16%) sedation (P = 0.554). Light sedation was not equivalent to deep sedation for procedural recall, the spectrum of complications, or recovery times. This study provides evidence to inform discussions with patients about sedation for colonoscopy. This trial was registered at the Australian and New Zealand Clinical Trials Registry, number 12611000320954.


Chronic disabling respiratory disease; ends and means of study.
In the program of research into the natural history of coalworkers' pneumoconiosis now being carried out in Great Britain, emphasis is being put upon the importance of sharpening and validating means for early diagnosis, and upon the need for follow-up studies upon properly selected population samples. Existing information from morbidity and mortality figures from chronic bronchitis and emphysema in Great Britain suggests that atmosphere pollutants are important etiological factors. A parallel is drawn between the course of events in pneumoconiosis, in which dust retention in the lungs does not greatly disable until complicated by tuberculous infection, and a hypothesis that bronchitis is a hypersecretion of bronchial mucus caused by atmospheric irritants and does not disable but encourages secondary infection which may cause emphysema. This hypothesis requires testing by follow-up studies of population samples exposed to various environmental influences. It is suggested that in order to bring this common and disabling disease under control, clinicians must widen their interests beyond the confines of the hospital walls.


Association of platelet counts on presentation and clinical outcomes in ST-elevation myocardial infarction (from the TIMI Trials).
Platelet activation and aggregation play pivotal roles in the thrombotic process of acute coronary syndromes. However, data regarding platelet count and its association with clinical outcomes in the setting of ST-elevation myocardial infarction (STEMI) are limited. We hypothesized that higher platelet counts on presentation would be associated with poorer clinical outcomes. Data from 10,793 patients with STEMI in the Thrombolysis In Myocardial Infarction (TIMI) trials database were analyzed. Mean platelet count on presentation was 254.8 x 10(3)/microl. Higher platelet counts were associated with higher rates of adverse clinical outcomes at 30 days. In a multivariable analysis that adjusted for confounders of platelet counts (age, gender, weight, diabetes, and smoking), higher platelet counts remained associated with an increased risk of the combined end point of death, reinfarction, and congestive heart failure. With a reference group of platelet counts <200 x 10(3)/microl, the multivariable odds ratios were 1.22 (95% confidence interval 1.05 to 1.42, p = 0.009) for platelet counts of 201 to 300 x 10(3)/microl, 1.37 (95% confidence interval 1.11 to 1.68, p = 0.002) for counts of 301 to 400 x 10(3)/microl, and 1.71 (95% confidence interval 1.16 to 2.51, p = 0.005) for counts >400 x 10(3)/microl. Further, a greater decrease in follow-up platelet counts (compared with baseline values) was independently associated with an increased risk of reinfarction at 30 days (odds ratio 1.44 for every decrease of 100 x 10(3)/microl unit of platelets, 95% confidence interval 1.13 to 1.82, p = 0.03). In conclusion, in STEMI, a higher platelet count on presentation was independently associated with adverse clinical outcomes, whereas a greater subsequent platelet count decrease was associated with an increased risk of reinfarction.


EEG correlates of categorical and graded face perception.
Face perception is a critical social ability and identifying its neural correlates is important from both basic and applied perspectives. In EEG recordings, faces elicit a distinct electrophysiological signature, the N170, which has a larger amplitude and shorter latency in response to faces compared to other objects. However, determining the face specificity of any neural marker for face perception hinges on finding an appropriate control stimulus. We used a novel stimulus set consisting of 300 images that spanned a continuum between random patches of natural scenes and genuine faces, in order to explore the selectivity of face-sensitive ERP responses with a model-based parametric stimulus set. Critically, our database contained "false alarm" images that were misclassified as face by computational face-detection system and varied in their image-level similarity to real faces. High-density (128-channel) event-related potentials (ERPs) were recorded while 23 adult subjects viewed all 300 images in random order, and determined whether each image was a face or non-face. The goal of our analyses was to determine the extent to which a gradient of sensitivity to face-like structure was evident in the ERP signal. Traditional waveform analyses revealed that the N170 component over occipitotemporal electrodes was larger in amplitude for faces compared to all non-faces, even those that were high in image similarity to faces, suggesting strict selectivity for veridical face stimuli. By contrast, single-trial classification of the entire waveform measured at the same sensors revealed that misclassifications of non-face patterns as faces increased with image-level similarity to faces. These results suggest that individual components may exhibit steep selectivity, but integration of multiple waveform features may afford graded information regarding stimulus appearance.


Molecular mapping of a histocompatibility-restricted immunodominant T cell epitope with synthetic and natural peptides: implications for T cell antigenic structure.
We have prepared synthetic and natural peptides that have allowed delineation of a major antigenic site of myoglobin recognized by histocompatibility (I-Ed)-restricted T cell clones. The smallest peptide capable of stimulating T cell proliferation consisted of residues 136-146. Residues Glu 136, Lys 140, and Lys 145 were essential for antigenicity, whereas Lys 133 and Tyr 146 added potency but were not required for antigenicity. The periodicity of these residues suggests that folding the peptide into its native alpha-helical structure may be essential for antigenicity either by forming a hydrophilic binding site for the T cell receptor or by participating in antigen presentation. The same folding could also produce a hydrophobic site on the opposite side of the alpha-helix that could participate in hydrophobic interactions. The extrapolation of these findings to other known peptide antigens suggests that this tendency to form an amphipathic alpha-helix may be a general property of antigenic sites recognized by T cells, perhaps due to a different functional role of each type of site in eliciting T cell responses.


A complex array of DNA-binding proteins required for pairing-sensitive silencing by a polycomb group response element from the Drosophila engrailed gene.
Regulatory DNA from the Drosophila gene engrailed causes silencing of a linked reporter gene (mini-white) in transgenic Drosophila. This silencing is strengthened in flies homozygous for the transgene and has been called "pairing-sensitive silencing." The pairing-sensitive silencing activities of a large fragment (2.6 kb) and a small subfragment (181 bp) were explored. Since pairing-sensitive silencing is often associated with Polycomb group response elements (PREs), we tested the activities of each of these engrailed fragments in a construct designed to detect PRE activity in embryos. Both fragments were found to behave as PREs in a bxd-Ubx-lacZ reporter construct, while the larger fragment showed additional silencing capabilities. Using the mini-white reporter gene, a 139-bp minimal pairing-sensitive element (PSE) was defined. DNA mobility-shift assays using Drosophila nuclear extracts suggested that there are eight protein-binding sites within this 139-bp element. Mutational analysis showed that at least five of these sites are important for pairing-sensitive silencing. One of the required sites is for the Polycomb group protein Pleiohomeotic and another is GAGAG, a sequence bound by the proteins GAGA factor and Pipsqueak. The identity of the other proteins is unknown. These data suggest a surprising degree of complexity in the DNA-binding proteins required for PSE function.


Possible tetramerisation of the proteasome maturation factor POMP/proteassemblin/hUmp1 and its subcellular localisation.
The proteasome is a multisubunit complex with a central role in non-lysosomal proteolysis and the processing of proteins for presentation by the MHC class I pathway. The 16kDa proteasome maturation protein POMP (also named proteassemblin or hUmp1) acts as a chaperone and is essential for the maturation of the 20S proteasome proteolytic core complex. However, the exact mechanism, timing and localisation of mammalian proteasome assembly remains elusive. We sought to investigate the localisation of POMP within the cell and therefore purified the protein and produced a polyclonal antibody. For immunisation, POMP was overexpressed and purified from a bacterial GST-system. Interestingly, after removal of the GST-tag, POMP was hardly detectable by Coomassie blue- and Ponceau red-staining. However, with a reverse zinc-staining, the protein could easily be visualised. POMP was gel-filtrated and eluted from a calibrated chromatography column with an apparent molecular weight of approximately 64kDa, suggesting that it forms tetramers. Moreover, localisation studies by immunofluorescence stainings and confocal microscopy revealed that POMP is present in the cytoplasm as well as in the nucleus.


Simplified representation of concepts and relations on screen.
The fully automated generation of diagnostic codes requires a knowledge-based system which is capable of interpreting noun phrases. The sense content of the words must be analysed and represented for this purpose. The codes are then generated based on this representation.In comparison with other knowledge-based systems, a system of this kind places the emphasis on the data structures and not on the calculus; coding itself is a simple matter compared to the much more difficult task of incorporating the complex information contained in the words used in natural language in a systematic data model. Initial attempts were based on the assumption that each word was linked to one conceptual meaning, whereas such a naive viewpoint certainly no longer applies today. The notation of concepts and their relations is the task at hand.Existing notation methods include predicate logic, conceptual graphs (CGs) as proposed by J. F. Sowa [2], GRAIL as used by the GALEN Project [1] and methods developed as part of the WWW consortium, e.g. RDF's (Resource Description Frameworks). For the purpose of coding, we developed a notation system using "concept particles" back in 1989 [3]. In 1996, the resulting experience led us to represent "concept molecules" (CM), with which both complex data structures and multi-branched rules can be denoted in a simple manner [4]. In this paper we shall explain the principles behind this notation and compare it with another modern concept representation system, conceptual graphs.


On paper synthesis of metal-organic framework as a chemiluminescence enhancer for estimating the total phenolic content of food samples using a smartphone readout.
Herein, a novel paper-based chemiluminescence (CL) assay is reported using a smartphone readout for on-site and reliable analytical applications. The CL system was based on the high-performance improving effect of cobalt-imidazole metal-organic framework (CoMOF) on luminol-hydrogen peroxide (H2O2) CL emission. The CoMOF was grown on paper and used as a support for the CL reaction, which led to an intense CL emission and good reproducibility. More importantly, the stability of luminol, as the CL reagent, was greatly improved in the presence of CoMOF. This high stability, along with the high-yield CL emission, makes the device highly suitable for commercialization. Furthermore, using a smartphone as the detector for the developed device made the process easier and more accessible for public usage. In this work, the new paper-based CL smartphone device was used for the detection of the total phenolic content of food samples. Phenolic compounds (PC) are hydroxyl radical scavengers that can effectively quench the CL emission of the luminol-H2O2-CoMOF system. After optimizing the reaction conditions, the system could detect PC at the μg mL-1 level. Detection limits of 0.12, 0.28, 0.46, 0.85, and 1.23 μg mL-1 were obtained for gallic acid, quercetin, catechin, kaempferol, and caffeic acid, respectively. This work is the first report on the practical application of smartphone CL assays for the estimation of PC. The proposed assay is an easy-to-use, low-cost, portable, and suitable assay for on-site screening purposes.


[p53 suppresses type II endometrial carcinomas in mice and governs endometrial tumor aggressiveness in humans].
Type II endometrial carcinomas are a highly aggressive group of tumor subtypes that are frequently associated with inactivation of the TP53 tumor suppressor gene. We show that mice with endometrium-specific deletion of the Trp53 gene initially exhibited histological changes that are identical to known precursor lesions of type II endometrial carcinomas in humans and later developed carcinomas representing all type II subtypes. The mTORC1 signalling pathway was frequently activated in these precursor lesions and tumors, suggesting a genetic cooperation between this pathway and Trp53 deficiency in tumor initiation. Consistent with this idea, analyses of 521 human endometrial carcinomas identified frequent mTORC1 pathway activation in type I as well as type II endometrial carcinoma subtypes. The mTORC1 pathway activation and p53 expression or mutation status each independently predicted poor patient survival. We suggest that molecular alterations in p53 and the mTORC1 pathway play different roles in the initiation of the different endometrial cancer subtypes but combined p53 inactivation and mTORC1 pathway activation are unifying pathogenic features among histologically diverse subtypes of late stage aggressive endometrial tumors.


Biotransformation of AFFF Component 6:2 Fluorotelomer Thioether Amido Sulfonate Generates 6:2 Fluorotelomer Thioether Carboxylate under Sulfate-Reducing Conditions.
The fate of per and polyfluoroalkyl substances (PFASs) in aqueous filmforming foams (AFFFs) under anaerobic conditions has not been well characterized, leaving major gaps in our understanding of PFAS fate and transformation at contaminated sites. In this study, the biotransformation of 6:2 fluorotelomer thioether amido sulfonate (6:2 FtTAoS), a component of several AFFF formulations, was investigated under sulfate-reducing conditions in microcosms inoculated with either pristine or AFFF-impacted solids. To identify the transformation products, we used high-resolution mass spectrometry and employed suspect-screening and nontargeted compound identification methods. These analyses demonstrated that 6:2 FtTAoS was transformed primarily to a stable polyfluoroalkyl compound, 6:2 fluorotelomer thioether propionate (6:2 FtTP). It did not undergo further reactions to produce the perfluoroalkyl carboxylates and fluorotelomer sulfonates and carboxylates that were observed during aerobic transformations. Here, the 6:2 FtTP was recalcitrant to biotransformation, indicating the stability of the thioether group under sulfate reducing conditions. The total oxidizable precursor (TOP) assay was used to assess the presence of other PFASs. Although nearly all of the PFAS mass initially present was recovered from the pristine microcosms, only 67% of the initial PFAS mass was recovered from the contaminated microcosms, suggesting the formation of volatile biotransformation products or those that could not be detected by the TOP assay.


Biosynthesis of the D2 cell adhesion molecule: pulse-chase studies in cultured fetal rat neuronal cells.
D2 is a membrane glycoprotein that is believed to function as a cell adhesion molecule (CAM) in neural cells. We have examined its biosynthesis in cultured fetal rat brain neurones. We found D2-CAM to be synthesized initially as two polypeptides: Mr 186,000 (A) and Mr 136,000 (B). With increasing chase times the Mr of both molecules increased to 187,000-201,000 (A) and 137,000-158,000 (B). These were similar to the sizes of D2-CAM labeled with [14C]glucosamine, [3H]fucose and [14C]mannosamine, indicating that the higher Mr species are glycoproteins. In the presence of tunicamycin, which specifically blocks the synthesis of high mannose cores, Mr were reduced to 175,000 (A) and 124,000 (B). Newly synthesized A and B are susceptible to degradation by endo-beta-N-acetyl-glucosaminidase H, which specifically degrades high mannose cores, but they are resistant to such degradation after 150 min of posttranslational processing. Hence, we deduce that A and B are initially synthesized with four to five high mannose cores which are later converted into N-linked complex oligosaccharides attached to asparagine residues. However, no shift of [35S]methionine radioactivity between A and B was detected with different pulse or chase times, showing that these molecules are not interconverted. Thus, our data indicate that the neuronal D2-CAM glycoproteins are derived from two mRNAs.


Presentation of obstructive left heart malformations in infancy.
This study documents the presentation of acutely life threatening congenital heart disease in infancy in order to assess the performance of screening examinations soon after birth and at 6 weeks of age. All cases of obstructive left heart malformations presenting in infancy in one health region from 1987-91 were analysed retrospectively. Altogether, 120 infants presented with either hypoplastic left heart syndrome, interruption of the aortic arch, coarctation of the aorta, or aortic valve stenosis. Twelve became symptomatic or died within 24 hours. Thirty four had an abnormal neonatal examination, eight of whom were referred. Six more became symptomatic before discharge. Ninety four babies went home, 51 developed heart failure before 6 weeks, and another seven died without diagnosis. Twenty five of 36 (69%) reaching 6 weeks without diagnosis had a second examination which was abnormal in 17. Two babies died undiagnosed after 6 weeks and the other 18 presented at up to 11 months of age. The neonatal and 6 week examinations perform poorly as screening tests for congenital heart disease. A normal neonatal examination does not exclude life threatening congenital heart disease.


Lipoxygenase metabolites of arachidonic and linoleic acids modulate the adhesion of tumor cells to endothelium via regulation of protein kinase C.
12(S)-hydroxyeicosatetraenoic acid (12[S]-HETE) and 13(S)-hydroxyoctadecadienoic acid (13[S]-HODE), lipoxygenase metabolites of arachidonic acid and linoleic acid, respectively, previously have been suggested to regulate tumor cell adhesion to endothelium during metastasis. Adhesion of rat Walker carcinosarcoma (W256) cells to a rat endothelial cell monolayer was enhanced after treatment with 12(S)-HETE and this 12(S)-HETE enhanced adhesion was blocked by 13(S)-HODE. Protein kinase inhibitors, staurosporine, calphostin C, and 1-(5-isoquinoline-sulfonyl)-2-methylpiperazine, inhibited the 12(S)-HETE enhanced W256 cell adhesion. Depleting W256 cells of protein kinase C (PKC) with phorbol 12-myristate-13-acetate abolished their ability to respond to 12(S)-HETE. Treatment of W256 cells with 12(S)-HETE induced a 100% increase in membrane-associated PKC activity whereas 13(S)-HODE inhibited the effect of 12(S)-HETE on PKC translocation. High-performance liquid chromatographic analysis revealed that in W256 cells 12-HETE and 13-HODE were two of the major lipoxygenase metabilites of arachidonic acid and linoleic acid, respectively. Therefore, these two metabolites may provide an alternative signaling pathway for the regulation of PKC. Further, these findings suggest that the regulation of tumor cell adhesion to endothelium by 12(S)-HETE and 13(S)-HODE may be a PKC-dependent process.


Developing public policy to advance the use of big data in health care.
The vast amount of health data generated and stored around the world each day offers significant opportunities for advances such as the real-time tracking of diseases, predicting disease outbreaks, and developing health care that is truly personalized. However, capturing, analyzing, and sharing health data is difficult, expensive, and controversial. This article explores four central questions that policy makers should consider when developing public policy for the use of "big data" in health care. We discuss what aspects of big data are most relevant for health care and present a taxonomy of data types and levels of access. We suggest that successful policies require clear objectives and provide examples, discuss barriers to achieving policy objectives based on a recent policy experiment in the United Kingdom, and propose levers that policy makers should consider using to advance data sharing. We argue that the case for data sharing can be won only by providing real-life examples of the ways in which it can improve health care.


Seletracetam (UCB 44212).
Better pharmacotherapies for epilepsy are needed for patients who are refractory to or have tolerability difficulties with current treatments. Seletracetam, a new drug in epilepsy development, is a pyrrolidone derivative structurally related to levetiracetam (trade name Keppra). It was discovered because of its high binding affinity to the synaptic vesicle 2A (SV2A) protein, which is now known to be the binding site for this family of compounds. Seletracetam shows very potent seizure suppression in models of acquired or genetic epilepsy, as well as high CNS tolerability in various animal models. Pharmacokinetic studies in animals suggest that seletracetam is rapidly and highly absorbed, with linear and time-independent pharmacokinetics. Seletracetam appears neither to inhibit nor to induce the major human drug metabolizing enzymes, and it demonstrates low plasma protein binding (<10%), which suggests a low potential for drug-drug interactions. Initial studies in humans demonstrated first-order monocompartmental kinetics with a half-life of 8 h and an oral bioavailability of >90%. Studies in healthy volunteers showed that the treatment emergent adverse events were of mild to moderate severity, were mostly of CNS origin and were resolved within 24 h. Altogether, these results suggest that seletracetam represents a promising new antiepileptic drug candidate, one that demonstrates a potent, broad spectrum of seizure protection and a high CNS tolerability in animal models, with initial clinical findings suggestive of straightforward pharmacokinetics and good tolerability.


A Rare Case of Hypersensitivity Pneumonitis due to Florida Red Tide.
Florida red tides occur annually due to proliferation of the marine dinoflagellate Karenia brevis, which produces neurotoxins known as brevotoxins. Inhalational exposure to brevotoxins usually results in upper airway symptoms only. Rarely does exposure lead to lower respiratory tract symptoms as in our case. We report a case of a 50-year-old man who presented with a 4-week history of dyspnea after exposure to the red tide. Computed tomography (CT) of the chest showed diffuse bilateral ground glass opacities and interstitial thickening. Bronchoalveolar lavage cultures and cytology were negative. The patient was started on steroids. Over the next few weeks, the patient's symptoms resolved. Repeat CT chest showed complete resolution of the ground glass opacities. Steroids were then tapered. Most patients who are exposed to algal blooms have self-limiting symptoms. Patients with asthma are particularly susceptible to worsening respiratory symptoms after exposure to brevotoxin aerosols. This case highlights that, in rare cases, exposure to red tide can results in severe lower respiratory tract symptoms.


Binding proteins for adenosine 3':5'-cyclic monophosphate in bovine adrenal cortex.
Five peaks of cyclic AMP-binding activity could be resolved by DEAE-cellulose chromatography of bovine adrenal-cortex cytosol. Two of the binding peaks co-chromatographed with the catalytic activities of cyclic AMP-dependent protein kinases (ATP-protein phosphotransferase, EC 126.96.36.199) of type I or type II respectively. A third binding protein was eluted between the two kinases, and appeared to be the free regulatory moiety of protein kinase I. Two of the binding proteins for cyclic AMP, sedimenting at 9S in sucrose gradients, could also bind adenosine. They bound cyclic AMP with an apparent equilibrium dissociation constant (K(d)) of about 0.1mum, and showed an increased binding capacity for cyclic AMP after preincubation in the presence of K(+), Mg(2+) and ATP. The two binding proteins differed in their apparent affinities for adenosine. The isolated regulatory moiety of protein kinase I had a very high affinity for cyclic AMP (K(d)<0.1nm). At low ionic strength or in the presence of MgATP, the high-affinity binding of cyclic AMP to the regulatory subunit of protein kinase I was decreased by the catalytic subunit. At high ionic strength and in the absence of MgATP the high-affinity binding to the regulatory subunit was not affected by the presence of catalytic subunit. Under all experimental conditions tested, dissociation of protein kinase I was accompanied by an increased affinity for cyclic AMP. To gain some insight into the mechanism by which cyclic AMP activates protein kinase, the interaction between basic proteins, salt and the cyclic nucleotide in activating the kinase was studied.


Modification of a small β-barrel protein, to give pseudo-amyloid structures, inhibits amyloid β-peptide aggregation.
Aggregation of amyloid β-peptide (Aβ) is closely related to the pathogenesis of Alzheimer's disease (AD). Although much effort has been devoted to the construction of molecules that inhibit the aggregation of Aβ1-42, high doses are needed for the inhibition of Aβ aggregation in many cases. Previously, we reported that designed green fluorescent protein (GFP) analogues that gives pseudo-Aβ β-sheet structures can work as an aggregation inhibitor against Aβ. To further test this design strategy, we constructed protein analogues that mimic Aβ β-sheet structures of amyloids by using insulin-like growth factor 2 receptor domain 11 (IGF2R-d11) as a scaffold. A designed protein, named IG11KK, which has a parallel configuration of Aβ-like β sheets, can bind more preferentially to oligomeric Aβ1-42 than the monomer. Moreover, IG11KK suppressed the aggregation of Aβ1-42 efficiently, even though lower concentrations of IG11KK than Aβ were used. The aggregation kinetics of Aβ in the presence of the designed proteins revealed that IG11KK can work as an inhibitor not only for the early to middle stages, but also in the latter stage of Aβ aggregation owing to its favorable binding to oligomeric structures of Aβ. The design strategy using β-barrel proteins such as IGF2R-d11 and GFP is useful in generating excellent inhibitors of protein misfolding and amyloid formation.


Communicating Risk: Developing an "Efficiency Index" for Dementia Screening Tests.
Diagnostic and screening tests may have risks such as misdiagnosis, as well as the potential benefits of correct diagnosis. Effective communication of this risk to both clinicians and patients can be problematic. The purpose of this study was to develop a metric called the "efficiency index" (EI), defined as the ratio of test accuracy and inaccuracy, to evaluate screening tests for dementia. This measure was compared with a previously described "likelihood to be diagnosed or misdiagnosed" (LDM), also based on "numbers needed" metrics. Datasets from prospective pragmatic test accuracy studies examining four brief cognitive screening instruments (Mini-Mental State Examination; Montreal Cognitive Assessment; Mini-Addenbrooke's Cognitive Examination (MACE); and Free-Cog) were analysed to calculate values for EI and LDM, and to examine their variation with test cut-off for MACE and dementia prevalence. EI values were also calculated using a modification of McGee's heuristic for the simplification of likelihood ratios to estimate percentage change in diagnostic probability. The findings indicate that EI is easier to calculate than LDM and, unlike LDM, may be classified either qualitatively or quantitatively in a manner similar to likelihood ratios. EI shows the utility or inutility of diagnostic and screening tests, illustrating the inevitable trade-off between diagnosis and misdiagnosis. It may be a useful metric to communicate risk in a way that is easily intelligible for both clinicians and patients.


First Published Report of Rust on White Alder Caused by Melampsoridium hiratsukanum in the United States.
White alder (Alnus rhombifolia) is a fast-growing tree native to the western United States and is planted frequently in landscapes. In September 2010, mature leaves of white alder with small, orange-yellow pustules were collected in a commercial nursery in Santa Cruz County, CA. Approximately 25 white alder trees were affected. Collected leaves were sent to the California Department of Food and Agriculture Plant Pest Diagnostics Laboratory. Young uredinial pustules were bullate, with urediniospores emerging from a single pore in the pustule. Spiny cells lined the ostiole. With age, pustules broke open to release more spores. Urediniospores were obovate to oval and measured from 14 to 20 × 27 to 41 μm (17.1 × 32.2 μm average, n = 62). Spores were uniformly echinulate and contained a nearly hyaline cell wall measuring from 1 to 2 μm (1.5 μm average) in thickness. A portion of the 28S ribosomal subunit (GenBank Accession No. KC313888) and the internal transcribed spacer regions (KC313889) were amplified and sequenced from DNA extracted from urediniospores using primers LR6 and rust2inv (1) and ITS1-F and ITS4-B (2), respectively. Our ITS sequence had 99% identity to GenBank accession EF564164, Melampsoridium hiratsukanum. In September 2011, white alder leaves with similar symptoms were collected from a commercial nursery in Santa Barbara County, CA. The spore morphology matched the white alder sample previously collected in Santa Cruz County, CA, in 2010. At that time, pathogenicity assays were conducted on three 1-year-old, 61-cm white alder trees planted in 3.8-liter pots. Six detached leaves with visible rust pustules were rubbed gently onto both the apical and distal side of moistened leaves of the healthy alders. Each infected leaf was used to inoculate a total of 6 to 10 healthy leaves by rubbing two leaves per tree before moving to the next tree. Leaves on three additional white alder trees were rubbed with healthy leaves as controls. Trees were incubated in a dew chamber for 3 days in darkness at 24°C, then placed in a growth chamber at 22°C with a 12-h photoperiod. Twelve days after inoculation, small lesions were visible on a few of the leaf undersides of each inoculated tree. Not all inoculated leaves developed pustules. No lesions developed on the control trees. M. hiratsukanum has been reported in Canada, Europe, and eastern Asia (3). There are no published reports of this rust in the United States, but there is an unpublished specimen from white alder in the USDA Systematic Mycology Herbarium (BPI 028048) collected from California in 1931, which was identified as M. hiratsukanum by G. B. Cummins using morphological criteria. We are unaware if older specimens of this rust exist because we were unable to search other herbaria in the United States. To the best of our knowledge, this rust has been present in California since 1931, but has only recently been found causing disease in nursery plants. There have been no reports of the serious foliar disease symptoms on trees in California wild lands as have been reported in Europe, presumably due to dry summer and fall seasons in white alder's natural habitat. References: (1) M. C. Aime. Mycoscience 47:112, 2006. (2) M. Gardes and T. D. Bruns. Mol. Ecol. 2:113, 1993. (3) J. Hatula et al. Mycologia 101:622, 2009.


Serial ricinine levels in serum and urine after ricin intoxication.
Ricinine is an alkaloid present in the castor bean plant (Ricinus communis) that can be used as a biomarker for ricin poisoning. Serial ricinine levels are reported in the serum and urine of a patient suffering from intentional ricin intoxication. The patient was brought to the hospital 4 h after injection and oral intake of a castor bean extract, but died 38 h later, despite intensive medical care. Ricinine was isolated from the samples by solid-phase extraction and quantitatively determined by isotopic dilution liquid chromatography-mass spectrometry. The ricinine level in serum declined from 33 to 23 ng/mL between 10 and 29 h post-exposure. Three urine samples collected from 12 to 41 h after ricin intoxication showed ricinine concentrations in the range of 20-58 ng/mL. The creatinine corrected values (21-30 µg/g) indicated a concentration-time profile with a maximum ricinine level in urine between 12 and 29 h after exposure.


An NAD+ Phosphorylase Toxin Triggers Mycobacterium tuberculosis Cell Death.
Toxin-antitoxin (TA) systems regulate fundamental cellular processes in bacteria and represent potential therapeutic targets. We report a new RES-Xre TA system in multiple human pathogens, including Mycobacterium tuberculosis. The toxin, MbcT, is bactericidal unless neutralized by its antitoxin MbcA. To investigate the mechanism, we solved the 1.8 Å-resolution crystal structure of the MbcTA complex. We found that MbcT resembles secreted NAD+-dependent bacterial exotoxins, such as diphtheria toxin. Indeed, MbcT catalyzes NAD+ degradation in vitro and in vivo. Unexpectedly, the reaction is stimulated by inorganic phosphate, and our data reveal that MbcT is a NAD+ phosphorylase. In the absence of MbcA, MbcT triggers rapid M. tuberculosis cell death, which reduces mycobacterial survival in macrophages and prolongs the survival of infected mice. Our study expands the molecular activities employed by bacterial TA modules and uncovers a new class of enzymes that could be exploited to treat tuberculosis and other infectious diseases.