Pediatric emergency department analgesic practice.
To review recent acute pain management care issues in a pediatric emergency department (ED) in order to identify opportunities for a performance improvement program. Descriptive, retrospective chart review. Urban pediatric hospital ED. Between January 1 and December 31, 1994 consecutive patients identified by ED chart review with the following three acute painful conditions were included; sickle cell vasoocclusive crisis (VOC) not complicated by fever or neurologic symptoms, isolated lower extremity long bone fractures < 12 hours old that did not require a reduction, and second degree burns < 12 hours old. Data collection concluded when between 50 and 55 episodes of each painful condition were identified. ED analgesic administration, initial analgesic dose, initial time elapsed to analgesic administration, notation of pain relief, and home analgesic instruction. Recommended analgesic starting doses were chosen from the 1992 Agency for Health Care Policy and Research Clinical Practice Guidelines. None. ED analgesic use for VOC was 100%, for fracture was 31%, and for burn was 26%. A recommended starting analgesic dose was given to 78% with VOC, 69% with fracture, and 79% with burn. Mean time to initial analgesic for VOC was 52 minutes, for fracture was 86 minutes, and for burn was 29 minutes. In those given analgesics, notation of pain relief for fracture was 19% and for burn was 29%, this improved for VOC where it was 88%. Home analgesic instruction for VOC was 100%, for fracture was 74%, and for burn was 27%. These data from 1994 document suboptimal analgesic use and home analgesic instruction for children in our ED with burns and fractures. Other opportunities in our ED for acute pain management improvement include optimizing initial analgesic doses, shortening the time elapsed to initial analgesic administration, and documenting the response to pain management.


Photochemical recovery of europium from non-aqueous solutions.
The photochemical recovery of europium from non-aqueous media, more specifically alcohols, is studied. The recovery was performed by photochemical reduction of europium(iii) to europium(ii) and subsequent removal as the insoluble EuCl2. Two charge transfer bands are present in the UV-C region, one originating from the alcohol (around 230 nm) and the other from the chloride anion (at 271 nm), which are responsible for the photochemical reduction when the solution is illuminated by a medium-pressure mercury lamp. When using different alcohol solvents, a trend is observed with regards to the removal rate and efficiency, following methanol (MeOH) < ethanol (EtOH) < isopropanol (IPA) <50/50 v/v ethanol/isopropanol (EtOH/IPA). This trend can be explained by the solubility of EuCl2 in the different solvents, and by the photon absorption at the wavelengths which provoke the reduction. In a 50/50 v/v EtOH/IPA solution, it is observed that addition of chloride ions (as LiCl) intensifies the chloride-to-europium(iii) CT band, effectively increasing the photon absorption in the 260-340 nm wavelength region. Moreover, addition of extra chloride ions decreases the solubility of EuCl2, which in turn accounts for a better recovery efficiency. However, this beneficial effect disappears when the water content rises above 1.5 wt%. For an EtOH/IPA solution with a high chloride concentration and low water content, it is feasible to recover europium from binary europium/yttrium mixtures with an efficiency of up to 94.7% and a purity of 96.7-99.8%, depending on the Eu/Y molar ratio. For higher yttrium excess, the removal rate of europium is higher, which is explained by the ability of yttrium to coordinate water molecules, decreasing the free water content in the solution. The fact that a large excess of yttrium does not compromise the removal rate of europium from the solution, proves that this technique has potential for europium recovery from red lamp phosphors (Y2O3:Eu3+), which consist entirely of europium and yttrium with a Eu/Y molar ratio of 1/20-1/30.


Hospital Transfer Network Structure as a Risk Factor for Clostridium difficile Infection.
To determine the effect of interhospital patient sharing via transfers on the rate of Clostridium difficile infections in a hospital. Retrospective cohort. Using data from the Healthcare Cost and Utilization Project California State Inpatient Database, 2005-2011, we identified 2,752,639 transfers. We then constructed a series of networks detailing the connections formed by hospitals. We computed 2 measures of connectivity, indegree and weighted indegree, measuring the number of hospitals from which transfers into a hospital arrive, and the total number of incoming transfers, respectively. Next, we estimated a multivariate model of C. difficile infection cases using the log-transformed network measures as well as covariates for hospital fixed effects, log median length of stay, log fraction of patients aged 65 or older, and quarter and year indicators as predictors. We found an increase of 1 in the log indegree was associated with a 4.8% increase in incidence of C. difficile infection (95% CI, 2.3%-7.4%) and an increase of 1 in log weighted indegree was associated with a 3.3% increase in C. difficile infection incidence (1.5%-5.2%). Moreover, including measures of connectivity in our models greatly improved their fit. Our results suggest infection control is not under the exclusive control of a given hospital but is also influenced by the connections and number of connections that hospitals have with other hospitals.


Photoluminescent nanographitic/nitrogen-doped graphitic hollow shells as a potential candidate for biological applications.
Water-dispersible graphitic hollow spheres were synthesized using a soft chemical route under hydrothermal conditions by glucose carbonization using a magnetite/silica-encapsulated core-shell sphere as a template. Carbonization on the templates happens as the magnetite core is partially or completely eliminated depending on the reaction conditions. Therefore, nano-sized graphitic hollow spheres or magnetite-core-encapsulated graphitic shells could be obtained. Also nitrogen-doped graphitic spheres were synthesized by a hydrothermal reaction. The graphitic and nitrogen-doped graphitic spheres show wavelength dependent photoluminescence in 300-600 nm range. The photoluminescence seems to depend on the fraction of the sp2 domains and N-doping, therefore, tunable PL emission can be achieved by controlling the nature of sp2 sites. In addition the cellular uptake of the graphitic hollow spheres was evaluated in human HeLa cells, demonstrating its main localization in the cytoplasm. A blue fluorescence signal was the most intensively observed in the cellular uptake process, although some green and red fluorescence was also observed. Since the cores of Fe3O4 could be completely or partly eliminated in a controllable way, it can be used as a magnetic resonance imaging agent. In addition, their easily modifiable hydrophilic surfaces for multi-functionality and hydrophobic voids covered by oxidized graphite make them promising candidates for applications in cellular photo-imaging and targeted drug delivery.


Prions: protein only or something more? Overview of potential prion cofactors.
Transmissible spongiform encephalopathies (TSEs) in humans and animals are attributed to protein-only infectious agents, called prions. Prions have been proposed to arise from the conformational conversion of the cellular protein PrP(C) into a misfolded form (e.g., PrP(Sc) for scrapie), which precipitates into aggregates and fibrils. It has been proposed that the conversion process is triggered by the interaction of the infectious form (PrP(Sc)) with the cellular form (PrP(C)) or might result from a mutation in the gene for PrP(C). However, until recently, all efforts to reproduce this process in vitro had failed, suggesting that host factors are necessary for prion replication. In this review we discuss recent findings such as the cellular factors that might be involved in the conformational conversion of prion proteins and the potential mechanisms by which they could operate.


Assessment of adherence to antiviral therapy in HIV-infected children using the Medication Event Monitoring System, pharmacy refill, provider assessment, caregiver self-report, and appointment keeping.
The authors sought to assess the utility of the electronic Medication Event Monitoring System (MEMS) in monitoring adherence to highly active antiretroviral therapy (HAART) in HIV-infected children and to compare this with other methods of adherence assessment. Twenty-six perinatally HIV-infected children being treated with three or more antiretroviral medications and their caregivers were enrolled and prospectively followed-up for 6 months. Adherence was assessed using MEMS monitoring of one antiretroviral, pharmacy refill records of all antiretrovirals, a caregiver self-report interview, a physician/nurse questionnaire, and appointment-keeping behavior. Viral loads measured at the end of the 6-month period were compared with the various adherence assessment methods. Adherence rates for the MEMS-monitored medication ranged from 12.7% to 97.9% (median = 81.4%), and 11 of the participants (42%) had less than 80% adherence using this method. A MEMS adherence rate greater than 80% was associated with viral load below the threshold of detection 6 months after enrollment (p <.001). Although not as robust, pharmacy refill rates for all antiretroviral medications were also associated with virologic response. The highest specificity was attained when both MEMS and pharmacy refill were used in combination. Physician assessment of adherence rate as well as appointment-keeping behavior was associated with virologic response, whereas caregiver self-report was not.


Current nutritional approaches in managing autism spectrum disorder: A review.
The link between nutrition and autism spectrum disorder (ASD), which is a complex developmental disorder manifesting itself in significant delays or deviation in interaction and communication, has provided a fresh point of view and signals that nutrition may have a role in the aetiology of ASD, as well as play an active role in treatment by alleviating symptoms. In this review study aimed at evaluating, with scientific and concrete proof, the current medical nutrition implementations on ASD, existing medical nutrition therapies have been addressed and their effects on ASD symptoms have been discussed in light of current research. We reviewed articles regarding the medical nutritional therapy of autism on current nutritional approaches selected from PubMed, Science Direct, EBSCO, and databases about autism and nutrition. The research put forward that in individuals with ASD, while gluten-free/casein-free and ketogenic diets, camel milk, curcumin, probiotics, and fermentable foods can play a role in alleviating ASD symptoms, consumption of sugar, additives, pesticides, genetically modified organisms, inorganic processed foods, and hard-to-digest starches may aggravate symptoms. Further prospective controlled trials with large sample sizes are needed before recommendations can be made regarding the ideal ASD diet. This review emphasizes the value of identifying current nutritional approaches specific to individuals with ASD and integrating their effects on symptoms to the conversation and presents suggestions for future research designed to identify medical nutrition therapies targeting this population to better understand the link between ASD and nutrition.


The TIMI risk score for unstable angina/non-ST elevation MI: A method for prognostication and therapeutic decision making.
Patients with unstable angina/non-ST-segment elevation myocardial infarction (MI) (UA/NSTEMI) present with a wide spectrum of risk for death and cardiac ischemic events. To develop a simple risk score that has broad applicability, is easily calculated at patient presentation, does not require a computer, and identifies patients with different responses to treatments for UA/NSTEMI. Two phase 3, international, randomized, double-blind trials (the Thrombolysis in Myocardial Infarction [TIMI] 11B trial [August 1996-March 1998] and the Efficacy and Safety of Subcutaneous Enoxaparin in Unstable Angina and Non-Q-Wave MI trial [ESSENCE; October 1994-May 1996]). A total of 1957 patients with UA/NSTEMI were assigned to receive unfractionated heparin (test cohort) and 1953 to receive enoxaparin in TIMI 11B; 1564 and 1607 were assigned respectively in ESSENCE. The 3 validation cohorts were the unfractionated heparin group from ESSENCE and both enoxaparin groups. The TIMI risk score was derived in the test cohort by selection of independent prognostic variables using multivariate logistic regression, assignment of value of 1 when a factor was present and 0 when it was absent, and summing the number of factors present to categorize patients into risk strata. Relative differences in response to therapeutic interventions were determined by comparing the slopes of the rates of events with increasing score in treatment groups and by testing for an interaction between risk score and treatment. Outcomes were TIMI risk score for developing at least 1 component of the primary end point (all-cause mortality, new or recurrent MI, or severe recurrent ischemia requiring urgent revascularization) through 14 days after randomization. The 7 TIMI risk score predictor variables were age 65 years or older, at least 3 risk factors for coronary artery disease, prior coronary stenosis of 50% or more, ST-segment deviation on electrocardiogram at presentation, at least 2 anginal events in prior 24 hours, use of aspirin in prior 7 days, and elevated serum cardiac markers. Event rates increased significantly as the TIMI risk score increased in the test cohort in TIMI 11B: 4.7% for a score of 0/1; 8.3% for 2; 13. 2% for 3; 19.9% for 4; 26.2% for 5; and 40.9% for 6/7 (P<.001 by chi(2) for trend). The pattern of increasing event rates with increasing TIMI risk score was confirmed in all 3 validation groups (P<.001). The slope of the increase in event rates with increasing numbers of risk factors was significantly lower in the enoxaparin groups in both TIMI 11B (P =.01) and ESSENCE (P =.03) and there was a significant interaction between TIMI risk score and treatment (P =. 02). In patients with UA/NSTEMI, the TIMI risk score is a simple prognostication scheme that categorizes a patient's risk of death and ischemic events and provides a basis for therapeutic decision making. JAMA. 2000;284:835-842


Are ultrasonography measurements of optic nerve sheath diameter an alternative to funduscopy in children with syndromic craniosynostosis?
Children with syndromic or complex craniosynostosis are evaluated for increased intracranial pressure (ICP) using funduscopy to detect papilledema. However, papilledema is a late sign of increased ICP. Because papilledema might be preceded by an increase in optic nerve sheath (ONS) diameter, the authors conducted a prospective study to establish the validity and applicability of measuring the ONS using ultrasonography. From January 2007 to December 2009, 175 bilateral ultrasonography ONS measurements were performed in 128 patients with syndromic or complex craniosynostosis during the daytime. The measurements were correlated with ONS diameter assessed on CT and simultaneous funduscopy, when available. Furthermore, results were compared by using thresholds for ONS diameters on ultrasonography that are available in the literature. The mean ONS diameter on ultrasonography was 3.1 ± 0.5 mm. The CT measurement was significantly correlated with the ultrasonography measurement (r = 0.41, p < 0.001). The mean ONS diameter in 38 eyes with papilledema was 3.3 ± 0.5 mm, compared with 3.1 ± 0.5 mm in the eyes of patients without papilledema (p = 0.039). Relative to the age-related thresholds, the ONS diameter was too large in 11 eyes (3%), particularly in patients with Crouzon syndrome. Compared with funduscopy, ultrasonography sensitivity was 11%, specificity was 97%, and positive and negative predictive values were 40% and 86%, respectively. Ultrasonography is a valid and easy way of quantifying the ONS. Although the ONS diameter is larger in children with papilledema, it cannot be used as a daytime screening tool instead of funduscopy. The ONS diameter is possibly a more real-time indicator of ICP.


Construction and biological analysis of deletion mutants of Fujinami sarcoma virus: 5'-fps sequence has a role in the transforming activity.
Fujinami sarcoma virus (FSV) genome codes for the gag-fps fusion protein FSV-P130. The amino acid sequence of the 3' one-third portion in v-fps is partially homologous to the 3' half of pp60src, or the kinase domain, but the sequence of the 5' portion is unique to v-fps. To identify a possible domain structure in the v-fps sequence responsible for cell transformation, we constructed various deletion mutants of FSV with molecularly cloned viral DNA. Their transforming activities were assayed by measuring focus formation on chicken embryo fibroblasts and rat 3Y1 cells and tumor formation in chickens. The mutants carrying a deletion at the 3' portion in v-fps, the kinase domain, lost transforming activity. The mutants carrying an approximately 1-kilobase deletion within the 5' portion of the v-fps sequence retained focus-forming activity and tumorigenicity in the chicken system, but the efficiency of focus formation was about 10 times lower than that of the wild type. The morphology of these transformed cells was distinct from that observed in cells infected with wild-type FSV. Furthermore, these mutants could not transform rat 3Y1 cells, although wild-type FSV DNA transformed rat 3Y1 cells at a high frequency. The mutants carrying a larger deletion in the 5' portion of fps completely lacked the transforming activity. These results suggest that the 3' portion of the v-fps sequence is necessary but not sufficient for cell transformation and that the 5' portion of v-fps has a role in the transforming activity.


Electric field directed self-assembly of cuprous oxide nanostructures for photon sensing.
We demonstrate a novel chemical-free water-based technique to synthesize various forms of cuprous oxide nanostructures at room temperature. The self-assemblies of these nanostructures are formed by the anodic oxidation of Cu in deionized (DI) water. Direct growth of these nanostructures on SiO(2)/Si (100) substrate has been successfully achieved by tuning the bias voltage and the growth duration. A variety of nanostructures from one-dimensional nanowires to different complex two- and three-dimensional structures are successfully grown by this method. We show that the morphological evolution in the self-assembly of the structures strongly depends on the spatial electric field distribution on the substrate. Furthermore, the electrical devices made from these nanowire networks exhibit promising photon sensing characteristics under white light illumination and can be exploited for future applications in photodetection and photovoltaic studies at the nanoscale level.


Development of common data elements: the experience of and recommendations from the early detection research network.
There have been an increasing number of large research consortia in recent years funded by the National Cancer Institute (NCI) to facilitate multi-disciplinary, multi-institutional cancer research. Some of these consortia have central data collection plans similar to a multi-center clinical trial whereas others plan to store data locally and pool or share the data at a later date. Regardless of the goal of the consortium, there is a need to standardize the way certain data are collected and stored, transferred, or reported across the institutions involved. This communication is a report of the process and current status of the development of common data elements (CDEs) by the Early Detection Research Network (EDRN). The development of the CDEs involved several stages with each stage requiring input from multi-disciplinary experts in oncology, epidemiology, biostatistics, pathology, informatics, and study coordination. An effort was made to be consistent with other consortia developing similar CDEs and to follow data standards when available. Initial focus was on identifying the minimum data that would be necessary to collect on all EDRN study participants and EDRN specimens. There are currently CDEs in the development or pilot phase for eight different organ sites and 13 different types of specimen procurements and plans to develop CDEs for 12 or more additional types of specimens.


Visual performance of myopia control soft contact lenses in non-presbyopic myopes.
To compare the visual performance of soft contact lenses reported to reduce myopia progression. In a double-blind, randomized, crossover trial, 30 non-presbyopic myopes wore MiSight™, center-distance Proclear® Multifocal (+2.00 D add), and two prototype lenses for 1 week each. High- and low-contrast visual acuities at 6 m, and 70 and 40 cm; stereopsis at 40 cm; accommodative facility at 33 cm; and horizontal phoria at 3 m and 33 cm were measured after 1 week. Subjective performance was assessed on a numeric rating scale for vision clarity, lack of ghosting, vision stability, haloes, overall vision satisfaction, and ocular comfort. Frequency of eye-strain symptoms and willingness to purchase lenses were also reported with categorical responses. Participants reported wearing times (total and visually acceptable). Linear mixed models and chi-square tests were employed in analysis with level of significance set at 5%. Theoretical optical performance of all lenses was assessed with schematic myopic model eyes (-1.00, -3.00, and -6.00 D) by comparing the slope of the edge spread function (ESF), an indicator for optical performance/resolution and the blur patch size of the line spread function, an indicator for contrast, between the lenses. Proclear Multifocal and MiSight provided the best distance acuities. However, the prototype lenses were rated significantly higher for many subjective variables, and there were no subjective variables where commercial lenses were rated significantly higher than the prototypes. Theoretical optical performance showed steeper slopes of the ESF and greater blur patch sizes of the LSP with commercial lenses, supporting the clinical findings of better visual acuities but reduced subjective performance. Participants wore prototypes longer and reported their vision acceptable for longer each day compared to MiSight. Both prototypes had the highest willingness-to-purchase rate. The prototypes were better tolerated by myopes compared to the commercial soft contact lenses currently used for slowing myopia progression.


Midsagittal magnetic resonance imaging of autism.
Since recent reports suggest structural brain abnormalities in autistic patients, we analysed magnetic resonance imaging (MRI) scans of autistic children. Planimetric measurements were done on midsagittal MRI scans, produced with a 0.5 T superconducting magnet. Scans of 13 'high-level' autistic subjects were compared with 35 control MRI scans, read as anatomically normal by a neuroradiologist. Corpus callosal, fourth ventricular, cerebellar, cerebral, and cranial areas were measured. The fourth ventricle was found to be significantly larger in the autistic group. No other areas in the midsagittal scans differed statistically between groups. Results suggest that structures defining the fourth ventricle are anatomically altered in autistic patients.


Improving quality of life by increasing outings after stroke: study protocol for the Out-and-About trial.
Almost one-third of Australians need help to travel outdoors after a stroke. Ambulation training and escorted outings are recommended as best practice in Australian clinical guidelines for stroke. Yet fewer than 20% of people with stroke receive enough of these sessions in their local community to change outcomes. The Out-and-About trial aims to determine the efficacy and cost effectiveness of an implementation program to change team behavior and increase outings by people with stroke. A two-group cluster-randomized trial will be conducted using concealed allocation, blinded assessors, and intention-to-treat analysis. Twenty community teams and their stroke clients (n = 300) will be recruited. Teams will be randomized to receive either the Out-and-About program or written guidelines only. The primary outcome is the proportion of people with stroke receiving multiple escorted outings during therapy sessions, measured at baseline and 13 months postintervention. Secondary outcomes include number of outings and distance traveled, measured using a self-report diary at baseline and six months postbaseline, and a global positioning system after six months. Cost effectiveness will measure quality-adjusted life years and health service use, measured at baseline and six months postbaseline. A potential outcome of this study will be evidence for a costed, transferable implementation program. If successful, the program will have international relevance and transferability. Another potential outcome will be validation of a novel and objective method of measuring outdoor travel (global positioning system) to supplement self-report methods.


Changes in hydration structure are necessary for collective motions of a multi-domain protein.
Conformational motions of proteins are necessary for their functions. To date, experimental studies measuring conformational fluctuations of a whole protein structure have revealed that water molecules hydrating proteins are necessary to induce protein functional motions. However, the underlying microscopic mechanism behind such regulation remains unsolved. To clarify the mechanism, multi-domain proteins are good targets because it is obvious that water molecules between domains play an important role in domain motions. Here, we show how changes in hydration structure microscopically correlate with large-amplitude motions of a multi-domain protein, through molecular dynamics simulation supported by structural analyses and biochemical experiments. We first identified collective domain motions of the protein, which open/close an active-site cleft between domains. The analyses on changes in hydration structure revealed that changes in local hydration in the depth of the cleft are necessary for the domain motion and vice versa. In particular, 'wetting'/'drying' at a hydrophobic pocket and 'adsorption'/'dissociation' of a few water molecules at a hydrophilic crevice in the cleft were induced by dynamic rearrangements of hydrogen-bond networks, and worked as a switch for the domain motions. Our results microscopically demonstrated the importance of hydrogen-bond networks of water molecules in understanding energy landscapes of protein motions.


Coexistence holes characterize the assembly and disassembly of multispecies systems.
A central goal of ecological research has been to understand the limits on the maximum number of species that can coexist under given constraints. However, we know little about the assembly and disassembly processes under which a community can reach such a maximum number, or whether this number is in fact attainable in practice. This limitation is partly due to the challenge of performing experimental work and partly due to the lack of a formalism under which one can systematically study such processes. Here, we introduce a formalism based on algebraic topology and homology theory to study the space of species coexistence formed by a given pool of species. We show that this space is characterized by ubiquitous discontinuities that we call coexistence holes (that is, empty spaces surrounded by filled space). Using theoretical and experimental systems, we provide direct evidence showing that these coexistence holes do not occur arbitrarily-their diversity is constrained by the internal structure of species interactions and their frequency can be explained by the external factors acting on these systems. Our work suggests that the assembly and disassembly of ecological systems is a discontinuous process that tends to obey regularities.


Empirical modelling of genetic algorithms.
This paper addresses the problem of reliably setting genetic algorithm parameters for consistent labelling problems. Genetic algorithm parameters are notoriously difficult to determine. This paper proposes a robust empirical framework, based on the analysis of factorial experiments. The use of a graeco-latin square permits an initial study of a wide range of parameter settings. This is followed by fully crossed factorial experiments with narrower ranges, which allow detailed analysis by logistic regression. The empirical models derived can be used to determine optimal algorithm parameters and to shed light on interactions between the parameters and their relative importance. Refined models are produced, which are shown to be robust under extrapolation to up to triple the problem size.


Relationship of distance run per week to coronary heart disease risk factors in 8283 male runners. The National Runners' Health Study.
Official guidelines from the Centers for Disease Control and Prevention and the American College of Sports Medicine state that every adult should accumulate 30 minutes or more of moderate-intensity physical activity on most, preferably all, days of the week. To examine the dose-response relationship between coronary heart disease (CHD) risk factors and vigorous exercise above the recommended minimum levels to assess whether further benefits accrue. Physician-supplied medical data were compared with reported distance run in a national cross-sectional survey of 8283 male recreational runners. Compared with runners who ran less than 16 km (10 miles) per week, long-distance runners (> or = 80 km/wk) showed an 85% reduced prevalence of high-density lipoprotein cholesterol levels that were clinically low (< 0.9 mmol/L [< 35 mg/dL]), a 2.5-fold increased prevalence of clinically defined high levels of high-density lipoprotein cholesterol (ie, > or = 1.55 mmol/L [> or = 60 mg/dL], the level thought to be protective against CHD), a nearly 50% reduction in hypertension, and more than a 50% reduction in the use of medications to lower blood pressure and plasma cholesterol levels. Estimated age-adjusted 10-year CHD risk was 30% lower in runners who averaged more than 64 km/wk than in those who averaged less than 16 km/wk (42 vs 61 events per 1000 men). Each 16-km incremental increase in weekly distance run up to 64 to 79 km/wk was associated with significant increases in high-density lipoprotein cholesterol levels and significant decreases in adiposity, triglyceride levels, the ratio of total cholesterol to high-density lipoprotein cholesterol level, and estimated CHD risk. Our data (1) suggest that substantial health benefits occur at exercise levels that exceed current minimum guidelines and (2) do not exhibit a point of diminishing return to the health benefits of running at any distance less than 80 km/wk.


Introduction of a norA promoter region mutation into the chromosome of a fluoroquinolone-susceptible strain of Staphylococcus aureus using plasmid integration.
It has been postulated that a mutation 11 bp 3' to the -10 motif of the norA promoter is involved in the increased expression of the gene observed in some strains of Staphylococcus aureus exhibiting efflux-related fluoroquinolone resistance. Introduction of this mutation into the chromosome of a fluoroquinolone-susceptible strain by plasmid integration resulted in the minimum inhibitory concentrations of NorA substrates being increased, fluoroquinolone uptake being reduced, and norA expression being enhanced. Diffuse hybridization of norA and integrating vector probes at a similar molecular weight range, higher than that of the norA transcript, was observed in the integrant, suggesting the possibility of a plasmid-based promoter contributing to norA expression. The ratio of the quantity of this transcript, which was also observed in the parent strain of the integrant, to the quantity of primary norA transcript was 0.14, demonstrating that it was unlikely that this mRNA species contributed significantly to the results observed. It is more likely that the introduced promoter region mutation does affect the expression of norA.


Using 1H and 13C NMR chemical shifts to determine cyclic peptide conformations: a combined molecular dynamics and quantum mechanics approach.
Solving conformations of cyclic peptides can provide insight into structure-activity and structure-property relationships, which can help in the design of compounds with improved bioactivity and/or ADME characteristics. The most common approaches for determining the structures of cyclic peptides are based on NMR-derived distance restraints obtained from NOESY or ROESY cross-peak intensities, and 3J-based dihedral restraints using the Karplus relationship. Unfortunately, these observables are often too weak, sparse, or degenerate to provide unequivocal, high-confidence solution structures, prompting us to investigate an alternative approach that relies only on 1H and 13C chemical shifts as experimental observables. This method, which we call conformational analysis from NMR and density-functional prediction of low-energy ensembles (CANDLE), uses molecular dynamics (MD) simulations to generate conformer families and density functional theory (DFT) calculations to predict their 1H and 13C chemical shifts. Iterative conformer searches and DFT energy calculations on a cyclic peptide-peptoid hybrid yielded Boltzmann ensembles whose predicted chemical shifts matched the experimental values better than any single conformer. For these compounds, CANDLE outperformed the classic NOE- and 3J-coupling-based approach by disambiguating similar β-turn types and also enabled the structural elucidation of the minor conformer. Through the use of chemical shifts, in conjunction with DFT and MD calculations, CANDLE can help illuminate conformational ensembles of cyclic peptides in solution.


Pacific healthcare workers and their treatment interventions for Pacific clients with alcohol and drug issues in New Zealand.
To provide an overview of the treatment interventions and practices of Pacific alcohol and other drugs (AOD) services in New Zealand. Face-to-face interviews were conducted with 31 Pacific staff members from 13 services registered with the Alcohol Advisory Council of New Zealand National Directory. Issues around assessment, treatment interventions, outcome measures, service structure, and resources were explored. Overall, the practices in District Health Board (DHB) and Non-Government Organisation (NGO) Pacific services were found to be similar. The clinical concepts of assessment, treatment, and outcome measures were not clearly understood by Pacific workers. This was due to the lack of attention towards Pacific concepts and practices and values. A holistic approach was viewed to be the best approach when working with Pacific clients based on health belief models, such as the Fonofale model. The findings of this study provide baseline data of treatment interventions and service practices of Pacific AOD services. There appears to be a Pacific way of working with Pacific clients. The most effective worker for Pacific people is someone who has sound knowledge of AOD, Pacific cultures and processes, and has the ability to integrate both Palangi (European) and Pacific knowledge to help the client.


The sequential appearance of low- and high-molecular-weight forms of MAP2 in the developing cerebellum.
Mammalian microtubule-associated protein 2 (MAP2) exists in high-molecular-weight (Mr approximately 280,000) and low-molecular-weight (Mr approximately 70,000) forms, with the latter protein being more abundant in embryonic brain homogenates than in preparations from mature brain (Riederer and Matus, 1985). In the current study, we have shown that avian MAP2 also exists as both high- (Mr approximately 260,000) and low-molecular-weight (Mr approximately 65,000) forms whose relative abundance changes during brain maturation, indicating a conserved function for these proteins during vertebrate neuronal morphogenesis. Using indirect immunohistochemistry, we have determined the cellular distribution of the high- and low-molecular-weight forms of MAP2 in the developing avian cerebellum. In the embryonic cerebellum, low-molecular-weight MAP2 is found in the external granular layer and in epithelial cells. High-molecular-weight MAP2 is found only in neurons that have commenced dendrogenesis, i.e., Purkinje cells and neurons within the internal granular layer. Thus, low-molecular-weight MAP2 is not only more abundant in embryonic nervous tissue than in the adult, but it also appears in glia and in differentiating neurons before the high-molecular-weight form. We have also shown that in the mature cerebellum high-molecular-weight MAP2 cannot be detected with monoclonal antibodies or polyclonal antisera in Purkinje cell dendrites. Polyclonal antisera against the regulatory subunit of the cAMP-dependent protein kinase, which is associated with MAP2 in the Purkinje cell dendrites of the rat, also fail to stain Purkinje cell dendrites in the mature quail cerebellum. This suggests that high-molecular-weight MAP2 may be necessary for the establishment of dendrites but is not necessary for the maintenance of dendritic form.


Priority rating for community care.
This paper, which is an additional nosokinetics paper to accompany those presented in Aust Health Rev 31(1), reports on priority rating through a standardised community care assessment system, based on screening for functional abilities and incorporating additional indicators of need and risk. Routinely collected measures used to generate a priority rating have proven useful in clinical decision making and active demand management at the service entry point. Priority rating is a step towards a more equitable and efficient assessment system. Three examples of priority rating systems are described. The first is a generalist application now implemented in routine practice across multiple service types in the Queensland community care and community health system. The second, narrower in scope, was designed for the NSW Home Care Service, and is also being routinely collected. The third was pilot tested in a state-wide program to supply aids and appliances to disabled people and introduced the additional concept of "capacity to benefit". The case studies show how a technical and data-driven approach can be useful in guiding policy in a complex health care sector.


Perturbed lignification impacts tree growth in hybrid poplar--a function of sink strength, vascular integrity, and photosynthetic assimilation.
The effects of reductions in cell wall lignin content, manifested by RNA interference suppression of coumaroyl 3'-hydroxylase, on plant growth, water transport, gas exchange, and photosynthesis were evaluated in hybrid poplar trees (Populus alba x grandidentata). The growth characteristics of the reduced lignin trees were significantly impaired, resulting in smaller stems and reduced root biomass when compared to wild-type trees, as well as altered leaf morphology and architecture. The severe inhibition of cell wall lignification produced trees with a collapsed xylem phenotype, resulting in compromised vascular integrity, and displayed reduced hydraulic conductivity and a greater susceptibility to wall failure and cavitation. In the reduced lignin trees, photosynthetic carbon assimilation and stomatal conductance were also greatly reduced, however, shoot xylem pressure potential and carbon isotope discrimination were higher and water-use efficiency was lower, inconsistent with water stress. Reductions in assimilation rate could not be ascribed to increased stomatal limitation. Starch and soluble sugars analysis of leaves revealed that photosynthate was accumulating to high levels, suggesting that the trees with substantially reduced cell wall lignin were not carbon limited and that reductions in sink strength were, instead, limiting photosynthesis.


Genetic architecture of Alzheimer's disease.
Advances in genetic and genomic technologies over the last thirty years have greatly enhanced our knowledge concerning the genetic architecture of Alzheimer's disease (AD). Several genes including APP, PSEN1, PSEN2, and APOE have been shown to exhibit large effects on disease susceptibility, with the remaining risk loci having much smaller effects on AD risk. Notably, common genetic variants impacting AD are not randomly distributed across the genome. Instead, these variants are enriched within regulatory elements active in human myeloid cells, and to a lesser extent liver cells, implicating these cell and tissue types as critical to disease etiology. Integrative approaches are emerging as highly effective for identifying the specific target genes through which AD risk variants act and will likely yield important insights related to potential therapeutic targets in the coming years. In the future, additional consideration of sex- and ethnicity-specific contributions to risk as well as the contribution of complex gene-gene and gene-environment interactions will likely be necessary to further improve our understanding of AD genetic architecture.


Assessment of Studies Evaluating Incremental Costs, Effectiveness, or Cost-Effectiveness of Systemic Therapies in Breast Cancer Based on Claims Data: A Systematic Review.
Large secondary databases, such as those containing insurance claims data, are increasingly being used to compare the effects and costs of treatments in routine clinical practice. Despite their appeal, however, caution must be exercised when using these data. In this study, we aimed to identify and assess the methodological quality of studies that used claims data to compare the effectiveness, costs, or cost-effectiveness of systemic therapies for breast cancer. We searched Embase, the Cochrane Library, Medline, Web of Science, and Google Scholar for English-language publications and assessed the methodological quality using the Good Research for Comparative Effectiveness principles. This study was registered with the International Prospective Register of Systematic Reviews (PROSPERO) under number CRD42018103992. We identified 1251 articles, of which 106 met the inclusion criteria. Most studies were conducted in the United States (74%) and Taiwan (9%) and were based on claims data sets (35%) or claims data linked to cancer registries (58%). Furthermore, most included large samples (mean 17 130 patients) and elderly patients, and they covered various outcomes (eg, survival, adverse events, resource use, and costs). Key methodological shortcomings were the lack of information on relevant confounders, the risk of immortal time bias, and the lack of information on the validity of outcomes. Only a few studies performed sensitivity analyses. Many comparative studies of cost, effectiveness, and cost-effectiveness have been published in recent decades based on claims data, and the number of publications has increased over time. Despite the availability of guidelines to improve quality, methodological issues persist and are often inappropriately addressed or reported.


Localized production of defence chemicals by intracellular symbionts of Haliclona sponges.
Marine sponges often house small-molecule-producing symbionts extracellularly in their mesohyl, providing the host with a means of chemical defence against predation and microbial infection. Here, we report an intriguing case of chemically mediated symbiosis between the renieramycin-containing sponge Haliclona sp. and its herein discovered renieramycin-producing symbiont Candidatus Endohaliclona renieramycinifaciens. Remarkably, Ca. E. renieramycinifaciens has undergone extreme genome reduction where it has lost almost all necessary elements for free living while maintaining a complex, multi-copy plasmid-encoded biosynthetic gene cluster for renieramycin biosynthesis. In return, the sponge houses Ca. E. renieramycinifaciens in previously uncharacterized cellular reservoirs (chemobacteriocytes), where it can acquire nutrients from the host and avoid bacterial competition. This relationship is highly specific to a single clade of Haliclona sponges. Our study reveals intracellular symbionts as an understudied source for defence chemicals in the oldest-living metazoans and paves the way towards discovering similar systems in other marine sponges.


Reaction of staphylococcal alpha-toxin with peptide-induced antibodies.
Two peptides representing separate 13-amino-acid sequences of staphylococcal alpha-toxin have been synthesized and acrylamide gel-purified alpha-toxin monomer and hexamer forms have been prepared and used to produce antisera in rabbits. We report here that each synthetic peptide, P-I and P-II, induces the formation of a specific precipitating antiserum. Moreover, these sera also react with the toxin monomer and sometimes with the hexamer, indicating that each peptide has more than one epitope. The purified toxin monomer can induce antibodies to fragments of toxin but is significantly less potent than the hexamer in inducing antibodies to the toxin monomer and almost not effective in inducing a response to the toxin hexamer. The purified toxin hexamer induces responses that are almost the reciprocals of the monomers, with the antihexamer and -monomer responses dominating and almost no responses to fragments of toxin being induced. These responses are interpreted in terms of the stability of the toxin hexamer to proteolytic degradation, compared with the relative sensitivity of the monomer to proteases. In assays of toxin-neutralization activity, only those sera containing antihexamer antibodies can block toxin hemolytic activity. This is true for both peptide- and toxin-induced antisera. The basis for this apparent association between toxin-neutralizing potency and antihexamer reactivity is being studied. Peptide P-I contains the uniquely reactive tyrosine residue and may be involved in monomer-to-monomer associations required to form hexamers. Peptide P-II is near the carboxyl terminus of alpha-toxin and may be involved in the binding of toxin to membranes. In a study of the ability of each peptide to inhibit the rate of hexamer formation induced by membrane lipoprotein, peptide P-I (as expected) proves to be more efficient than peptide P-II. Finally, one rabbit immunized with the toxin hexamer produces antibodies to peptides P-I and P-II. This finding suggests that the two synthetic peptides selected for study are relevant to the in vivo immunoprocessing of staphylococcal alpha-toxin.


Influence of local structure on photoluminescence properties of Eu3+ doped CeO2 red phosphors through induced oxygen vacancies by contrasting rare earth substitutions.
A new family of red phosphors, Ce0.9-xRExO2-δ:0.1Eu3+ (RE = Y and La; x = 0, 0.20, 0.40, 0.60, 0.80, and 0.90), was synthesized by a conventional solid-state route. The influence of contrasting rare earth substitutions (Y and La) in the system was investigated on the local structure and associated photoluminescence properties by various characterization techniques. Both trivalent ion substitutions lead to the same kind of variation during phase transformation from fluorite to the respective parent oxide structure (x ≥ 0.6). On the other hand, the substitutions have a distinct effect on local structure, absorption, luminescence and lifetimes. The smaller Y3+ ion substitution enables the ordering of oxygen vacancies in a lattice, inhibiting the defect formation of Ce3+ oxidation states. In contrast, the larger La3+ ion substitution liberates oxygen vacancies, allowing defect formation. Consequently, the concentration of Ce3+ is dependent on the ionic radius of the metal ions and it has a bearing on the band gap and luminescence properties of the system. Ce0.1Y0.8O2-δ:0.1Eu3+ phosphor exhibited maximum red emission intensity at 612 nm, which is 8 times higher than that of Ce0.9O2-δ:0.1Eu3+ and better than that of commercial Philips red phosphor, whereas La substitution yielded poor emission intensities with higher concentrations. The co-substitution of contrasting rare earth metals with Eu3+ allow the understanding of local structure and a smaller ion like Y3+ greatly functionalizes CeO2:Eu3+ phosphor.


cAMP-dependent protein kinase A and the dynamics of epithelial cell surface domains: moving membranes to keep in shape.
Cyclic adenosine monophosphate (cAMP) and cAMP-dependent protein kinase A (PKA) are evolutionary conserved molecules with a well-established position in the complex network of signal transduction pathways. cAMP/PKA-mediated signaling pathways are implicated in many biological processes that cooperate in organ development including the motility, survival, proliferation and differentiation of epithelial cells. Cell surface polarity, here defined as the anisotropic organisation of cellular membranes, is a critical parameter for most of these processes. Changes in the activity of cAMP/PKA elicit a variety of effects on intracellular membrane dynamics, including membrane sorting and trafficking. One of the most intriguing aspects of cAMP/PKA signaling is its evolutionary conserved abundance on the one hand and its precise spatial-temporal actions on the other. Here, we review recent developments with regard to the role of cAMP/PKA in the regulation of intracellular membrane trafficking in relation to the dynamics of epithelial surface domains.


The Association between Bisphenol A Exposure and Obesity in Children-A Systematic Review with Meta-Analysis.
Bisphenol A (BPA) is an environmental chemical that has adverse effects on health, probably causing childhood obesity. However, this association remains controversial, and it is difficult to find evidence for direct causality between environmental exposure and disease using epidemiological studies. In this study, we sought to elucidate the possible causality between BPA exposure and childhood obesity by conducting two meta-analyses showing bidirectional associations, including exposure effect by obesity and obesity risk by exposure. Articles published up to September 2017 were searched in PubMed, Embase, and Cochrane Library. We evaluated observational studies that included measurements of urinary BPA concentration and BMI or body weight. Of 436 articles, a total of 13 studies were included in the meta-analysis. Two meta-analyses were performed to investigate the association between BPA exposure and childhood obesity. The results showed that the relatively high-exposed group had a significantly higher risk of childhood obesity than the relatively low-exposed group (odds ratio = 1.566, 95% confidence interval [CI]: 1.097 to 2.234, p = 0.014). However, the obese group showed no significant difference in the BPA concentration when compared to the normal group (standardized mean difference = 0.166, 95% CI: -0.121 to 0.453, p = 0.257). This study suggested possible causality between BPA exposure and childhood obesity using data from epidemiological studies and showed that BPA exposure itself increased the risk of obesity in children.


A Randomized trial of an Asthma Internet Self-management Intervention (RAISIN): study protocol for a randomized controlled trial.
The financial costs associated with asthma care continue to increase while care remains suboptimal. Promoting optimal self-management, including the use of asthma action plans, along with regular health professional review has been shown to be an effective strategy and is recommended in asthma guidelines internationally. Despite evidence of benefit, guided self-management remains underused, however the potential for online resources to promote self-management behaviors is gaining increasing recognition. The aim of this paper is to describe the protocol for a pilot evaluation of a website 'Living well with asthma' which has been developed with the aim of promoting self-management behaviors shown to improve outcomes. The study is a parallel randomized controlled trial, where adults with asthma are randomly assigned to either access to the website for 12 weeks, or usual asthma care for 12 weeks (followed by access to the website if desired). Individuals are included if they are over 16-years-old, have a diagnosis of asthma with an Asthma Control Questionnaire (ACQ) score of greater than, or equal to 1, and have access to the internet. Primary outcomes for this evaluation include recruitment and retention rates, changes at 12 weeks from baseline for both ACQ and Asthma Quality of Life Questionnaire (AQLQ) scores, and quantitative data describing website usage (number of times logged on, length of time logged on, number of times individual pages looked at, and for how long). Secondary outcomes include clinical outcomes (medication use, health services use, lung function) and patient reported outcomes (including adherence, patient activation measures, and health status). Piloting of complex interventions is considered best practice and will maximise the potential of any future large-scale randomized controlled trial to successfully recruit and be able to report on necessary outcomes. Here we will provide results across a range of outcomes which will provide estimates of efficacy to inform the design of a future full-scale randomized controlled trial of the 'Living well with asthma' website. This trial is registered with Current Controlled Trials ISRCTN78556552 on 18/06/13.


Signal integration and the specificity of insulin action.
Insulin is a potent metabolic hormone essential for the maintenance of normal circulating blood glucose level in mammals. The physiologic control of glucose homeostasis results from a balance between hepatic glucose release (glycogenolysis and gluconeogenesis) and dietary glucose absorption versus skeletal muscle and adipose tissue glucose uptake and disposal. Disruption of this delicate balance either through defects in insulin secretion, liver glucose output, or peripheral tissue glucose uptake results in pathophysiological states of insulin resistance and diabetes. In particular, glucose transport into skeletal muscle and adipose tissue is the rate-limiting step in glucose metabolism and reduction in the efficiency of this process (insulin resistance) is one of the earliest predictors for the development of Type II diabetes. Importantly, recent studies have directly implicated an impairment in insulin receptor signal transduction as the prime mechanism for peripheral tissue insulin resistance. In this review, we have focused on recent developments in our understanding of the molecular mechanisms and signal transduction pathways that insulin utilizes to specifically regulate glucose uptake. The detailed understanding of these events will provide a conceptual framework for the development of new therapeutic targets to treat this chronic and debilitating disease process.


Effect of aprotinin on clinical outcomes in coronary artery bypass graft surgery: a systematic review and meta-analysis of randomized clinical trials.
Despite proven blood transfusion benefits, aprotinin may be underused in coronary artery bypass grafting. Reluctance to use aprotinin may stem from safety concerns. The current objective was to evaluate clinical outcomes (mortality, myocardial infarction, renal failure, stroke, atrial fibrillation) in patients undergoing coronary artery bypass grafting who receive aprotinin by performing a quantitative overview of published, randomized, controlled trials. MEDLINE, EMBASE, and PHARMLINE (1988-2001) and reference lists of relevant articles were searched for coronary artery bypass grafting studies. Criteria for data inclusion were as follows: (1) random allocation of study treatments, (2) placebo control, (3) enrollment only of patients undergoing coronary artery bypass grafting, (4) no combination with another experimental medication or device, and (5) prophylactic and continuous intraoperative use. Data from 35 coronary artery bypass grafting trials (n = 3879) confirm that aprotinin reduces transfusion requirements (relative risk 0.61, 95% confidence interval 0.58-0.66) relative to placebo, with a 39% risk reduction. Aprotinin therapy was not associated with increased or decreased mortality (relative risk 0.96, 95% confidence interval 0.65-1.40), myocardial infarction (relative risk 0.85, 95% confidence interval 0.63-1.14), or renal failure (relative risk 1.01, 95% confidence interval 0.55-1.83) risk, but it was associated with a reduced risk of stroke (relative risk 0.53, 95% confidence interval 0.31-0.90) and a trend toward reduced atrial fibrillation (relative risk 0.90, 95% confidence interval 0.78-1.03). Aprotinin reduces transfusion requirements. Concerns that aprotinin therapy is associated with increased mortality, myocardial infarction, or renal failure risk is not supported by data from published, randomized, placebo-controlled clinical trials. Evidence for a reduced risk of stroke and a tendency toward reduction of atrial fibrillation occurrence was observed in patients who received aprotinin.


Association between herbivore stress and glutathione S-transferase expression in Pinus brutia Ten.
Plants have developed mechanisms to defend themselves against many factors including biotic stress such as herbivores and pathogens. Glutathione S-transferase (GST) is a glutathione-dependent detoxifying enzyme and plays critical roles in stress tolerance and detoxification metabolism in plants. Pinus brutia Ten. is a prominent native forest tree species in Turkey, due to both its economic and ecological assets. One of the problems faced by P. brutia afforestation sites is the attacks by pine processionary moth (Thaumetopoea wilkinsoni Tams.). In this study, we investigated the changes in activity and mRNA expression of GST in pine samples taken from both resistant and susceptible clones against T. wilkinsoni over a nine month period in a clonal seed orchard. It was found that the average cytosolic GST activities of trees in March and July were significantly higher than the values obtained in November. November was considered to be the control since trees were not under stress yet. In addition, RT-PCR results clearly showed that levels of GST transcripts in March and July samples were significantly higher as compared to the level seen in November. These findings strongly suggest that GST activity from P. brutia would be a valuable marker for exposure to herbivory stress.


The presence and response to Zn of ZnT family mRNAs in human dental pulp.
Zinc (Zn) is distributed throughout the body and within cells by saturable processes mediated by the transport proteins of the ZnT (SLC30) and ZIP (SLC39) families. The two families function in opposite directions. ZnT transporters mediate cellular zinc efflux or intracellular sequestration. Zn is found in human tooth enamel and dentine at levels that have been related to environmental exposures, such as pollution, disease, and dietary intake. The mechanism by which Zn in the odontoblast is deposited in the hard tissue of the tooth, however, is unknown but is important in determining the physical properties, and hence resilience, of enamel and in the context of the use of tooth zinc level as a biomarker of exposure. We hypothesised that zinc efflux mediated by members of the ZnT family of 10 transporters is a key step in this process and is regulated by zinc availability through effects on mRNA levels. Thus, we determined the profile of ZnT transporter mRNA in a human active-secretory odontoblast-like cell model under conditions of high- and low-extracellular Zn concentration and determined if the same transporter mRNAs were present in human dental pulp. ZnT1, ZnT5 and ZnT9 mRNAs were detected by RT-PCR in both the secretory odontoblast cells and human dental pulp. ZnT2, ZnT3 and ZnT10 mRNAs were not detected, and ZnT4 mRNA was detected in secretory odontoblasts only, which may be indicative of a specialised zinc efflux function during the active secretory phase of tooth development. ZnT1 mRNA was significantly increased in response to extracellular Zn exposure (60 μM) after 24 h. The presence of Zn transporter mRNAs in secretory odontoblasts and dental pulp indicates that the corresponding transport proteins function to deposit zinc in the dental hard tissues. The responsiveness of ZnT1 in odontoblasts to zinc availability is concordant with this being a process that is regulated to maintain cellular Zn homeostasis and that is a mediator of the relationship between environmental Zn exposure and dental Zn deposition. These findings have likely relevance to human dental health through effects of Zn transporter expression level on the hard tissue properties.


Reproducibility of Structural and Diffusion Tensor Imaging in the TACERN Multi-Center Study.
Multi-site MRI studies are often necessary for recruiting sufficiently sized samples when studying rare conditions. However, they require pooling data from multiple scanners into a single data set, and therefore it is critical to evaluate the variability of quantitative MRI measures within and across scanners used in multi-site studies. The aim of this study was to evaluate the reproducibility of structural and diffusion weighted (DW) MRI measurements acquired on seven scanners at five medical centers as part of the Tuberous Sclerosis Complex Autism Center of Excellence Research Network (TACERN) multisite study. The American College of Radiology (ACR) phantom was imaged monthly to measure reproducibility of signal intensity and uniformity within and across seven 3T scanners from General Electric, Philips, and Siemens vendors. One healthy adult male volunteer was imaged repeatedly on all seven scanners under the TACERN structural and DW protocol (5 b = 0 s/mm2 and 30 b = 1000 s/mm2) over a period of 5 years (age 22-27 years). Reproducibility of inter- and intra-scanner brain segmentation volumes and diffusion tensor imaging metrics fractional anisotropy (FA) and mean diffusivity (MD) within white matter regions was quantified with coefficient of variation. The American College of Radiology Phantom signal intensity and uniformity were similar across scanners and changed little over time, with a mean intra-scanner coefficient of variation of 3.6 and 1.8%, respectively. The mean inter- and intra-scanner coefficients of variation of brain structure volumes derived from T1-weighted (T1w) images of the human phantom were 3.3 and 1.1%, respectively. The mean inter- and intra-scanner coefficients of variation of FA in white matter regions were 4.5 and 2.5%, while the mean inter- and intra-scanner coefficients of variation of MD in white matter regions were 5.4 and 1.5%. Our results suggest that volumetric and diffusion tensor imaging (DTI) measurements are highly reproducible between and within scanners and provide typical variation amplitudes that can be used as references to interpret future findings in the TACERN network.


A matched case-control study of factors associated with retention of fetal membranes in dairy ewes in Southern Greece.
We defined retention of fetal membranes (RFM) in dairy ewes as failure to expel the placenta within 6h after lambing the last lamb and designed a matched case-control study to identify factors that affect the risk of retention. For each ewe with RFM, the next ewe in the flock that lambed and expelled the placenta in <6h after lambing the last lamb was selected as control. Data analyzed included 92 pairs of ewes from 25 flocks comprising a total of 7275 ewes (median flocksize 270 ewes). Factors investigated for associations with RFM were induction of lambing, obstetrical assistance because of dystocia, parity, the number of liveborn lambs, the occurrence of stillbirth(s), the mean weight of the litter on the third day post-lambing and the occurrence of neonatal death in the litter. Conditional logistic regression indicated (1) that the risk of RFM increased linearly with increasing number of liveborn lambs and (2) that the risk of retention was 4-fold higher in ewes that received assistance at lambing than those that lambed normally.


Alanine scanning analyses of the three major loops in domain II of Bacillus thuringiensis mosquitocidal toxin Cry4Aa.
Cry4Aa produced by Bacillus thuringiensis is a dipteran-specific toxin and is of great interest for developing a bioinsecticide to control mosquitoes. Therefore, it is very important to characterize the functional motif of Cry4Aa that is responsible for its mosquitocidal activity. In this study, to characterize a potential receptor binding site, namely, loops 1, 2, and 3 in domain II, we constructed a series of Cry4Aa mutants in which a residue in these three loops was replaced with alanine. A bioassay using Culex pipiens larvae revealed that replacement of some residues affected the mosquitocidal activity of Cry4Aa, but the effect was limited. This finding was partially inconsistent with previous results which suggested that replacement of the Cry4Aa loop 2 results in a significant loss of mosquitocidal activity. Therefore, we constructed additional mutants in which multiple (five or six) residues in loop 2 were replaced with alanine. Although the replacement of multiple residues also resulted in some decrease in mosquitocidal activity, the mutants still showed relatively high activity. Since the insecticidal spectrum of Cry4Aa is specific, Cry4Aa must have a specific receptor on the surface of the target tissue, and loss of binding to the receptor should result in a complete loss of mosquitocidal activity. Our results suggested that, unlike the receptor binding site of the well-characterized molecule Cry1, the receptor binding site of Cry4Aa is different from loops 1, 2, and 3 or that there are multiple binding sites that work cooperatively for receptor binding.


Sensitization of lung cancer cells to cisplatin by β-elemene is mediated through blockade of cell cycle progression: antitumor efficacies of β-elemene and its synthetic analogs.
The development of effective agents for overcoming platinum chemoresistance in lung carcinoma continues to have high priority. We have demonstrated recently that β-elemene, a novel antitumor compound, enhances cisplatin activity by triggering lung cancer cell death via apoptosis. Here, we investigated whether β-elemene acts synergistically with cisplatin to inhibit non-small cell lung cancer (NSCLC) cell proliferation by blocking cell cycle progression. β-Elemene substantially increased the suppressive effect of cisplatin on cell growth and proliferation in the NSCLC cell lines H460 and A549. Furthermore, β-elemene augmented cisplatin in the cell cycle arrest of NSCLC cells at G(2)/M. This was associated with upregulated checkpoint kinase (CHK2) expression and reduced CDC2 activity (i.e., increased phosphorylation of CDC2 on Tyr-15 and decreased phosphorylation of CDC2 on Thr-161). Moreover, β-elemene and cisplatin in combination clearly decreased the protein levels of cyclin B1 and CDC25C and increased the levels of p21(Cip1/Waf1), p27(Kip1), and GADD45 in these cells, compared with the effects of either agent alone at the same concentration. These results suggest that the β-elemene-enhanced inhibitory effect of cisplatin on lung carcinoma cell proliferation is regulated by a CHK2-mediated CDC25C/CDC2/cyclin B1 signaling pathway and leads to the blockade of cell cycle progression at G(2)/M. A comparison of the cytotoxic efficacies of β-elemene and three synthetic analogs (β-elemenol, β-elemenal, and β-elemene fluoride) in the two lung cancer cell lines revealed that β-elemenol and β-elemene fluoride had the same antitumor efficacy as β-elemene, whereas β-elemenal was appreciably more potent than β-elemene. Thus, although all three synthetic analogs of β-elemene considerably suppressed NSCLC cell growth and proliferation, β-elemenal may have greater potential as an anticancer alternative to β-elemene in treating lung cancer and other tumors.


Human amnion epithelial cells induced to express functional cystic fibrosis transmembrane conductance regulator.
Cystic fibrosis, an autosomal recessive disorder caused by a mutation in a gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR), remains a leading cause of childhood respiratory morbidity and mortality. The respiratory consequences of cystic fibrosis include the generation of thick, tenacious mucus that impairs lung clearance, predisposing the individual to repeated and persistent infections, progressive lung damage and shortened lifespan. Currently there is no cure for cystic fibrosis. With this in mind, we investigated the ability of human amnion epithelial cells (hAECs) to express functional CFTR. We found that hAECs formed 3-dimensional structures and expressed the CFTR gene and protein after culture in Small Airway Growth Medium (SAGM). We also observed a polarized CFTR distribution on the membrane of hAECs cultured in SAGM, similar to that observed in polarized airway cells in vivo. Further, hAECs induced to express CFTR possessed functional iodide/chloride (I(-/)Cl(-)) ion channels that were inhibited by the CFTR-inhibitor CFTR-172, indicating the presence of functional CFTR ion channels. These data suggest that hAECs may be a promising source for the development of a cellular therapy for cystic fibrosis.


Recovery rate and associated factors of children age 6 to 59 months admitted with severe acute malnutrition at inpatient unit of Bahir Dar Felege Hiwot Referral hospital therapeutic feeding unite, northwest Ethiopia.
Despite numerous advances made in improving child health and the clinical management protocols for treating severe acute malnutrition at treatment centers, evidences concerning the treatment outcomes are scarce. Therefore, this study was conducted to assess the recovery rate and associated factors of severely acute malnourished children of age 6 to 59 months admitted to inpatient therapeutic feeding unit at Felege Hiwot Referral Hospital. We conducted a hospital-based cross-sectional study including 401 severely malnourished children who were admitted from September 2012 to January 2016. Bivariable and a Multivariable logistic regression model were fitted to identify factors associated with recovery rate. Adjusted Odds ratio with its 95% CI was reported and P-value less than 0.05 was considered as significant. Fifty eight percent (58.4%) (95%CI: 53.1-64.1) of admitted children were recovered with a mean recovery time of 18 (±6.3) days. Being female, children who were fully and partially vaccinated, who had better MUAC measurement, who stayed longer in the hospital, and children who took routine vitamin-A supplementation had better recovery rate. However, children who had co-morbidity at admission, had human immune virus (HIV) and Tuberculosis (TB) infection, and who had edema were less likely to recover. Recovery rate was low as compared to international SPHERE cutoff points (> 75% recovery rate). Interventions that could address the outlined factors would be helpful to improve treatment recovery rate of admitted children.


An enhanced regimen as post-exposure chemoprophylaxis for leprosy: PEP+.
The ongoing transmission of Mycobacterium (M.) leprae reflected in a very slow decline in leprosy incidence, forces us to be innovative and conduct cutting-edge research. Single dose rifampicin (SDR) as post-exposure prophylaxis (PEP) for contacts of leprosy patients, reduces their risk to develop leprosy by 60%. This is a promising new preventive measure that can be integrated into routine leprosy control programmes, as is being demonstrated in the Leprosy Post-Exposure Programme that is currently ongoing in eight countries.The limited (60%) effectiveness of SDR is likely due to the fact that some contacts have a preclinical infection beyond the early stages for which SDR is not sufficient to prevent the development of clinical signs and symptoms of leprosy. An enhanced regimen, more potent against a higher load of leprosy bacteria, would increase the effectiveness of this preventive measure significantly.The Netherlands Leprosy Relief (NLR) is developing a multi-country study aiming to show that breaking the chain of transmission of M. leprae is possible, evidenced by a dramatic reduction in incidence. In this study the assessment of the effectiveness of an enhanced prophylactic regimen for leprosy is an important component. To define the so called PEP++ regimen for this intervention study, NLR convened an Expert Meeting that was attended by clinical leprologists, public health experts, pharmacologists, dermatologists and microbiologists.The Expert Meeting advised on combinations of available drugs, with known efficacy against leprosy, as well as on the duration of the intake, aiming at a risk reduction of 80-90%. To come to a conclusion the Expert Meeting considered the bactericidal, sterilising and bacteriostatic activity of the potential drugs. The criteria used to determine an optimal enhanced regimen were: effectiveness, safety, acceptability, availability, affordability, feasibility and not inducing drug resistance.The Expert Meeting concluded that the enhanced regimen for the PEP++ study should comprise three standard doses of rifampicin 600 mg (weight adjusted when given to children) plus moxifloxacin 400 mg given at four-weekly intervals. For children and for adults with contraindications for moxifloxacin, moxifloxacin should be replaced by clarithromycin 300 mg (weight adjusted).


RNA sequence and two-dimensional structure features required for efficient substrate modification by the Saccharomyces cerevisiae RNA:{Psi}-synthase Pus7p.
The RNA:pseudouridine (Psi) synthase Pus7p of Saccharomyces cerevisiae is a multisite-specific enzyme that is able to modify U(13) in several yeast tRNAs, U(35) in the pre-tRNA(Tyr) (GPsiA), U(35) in U2 small nuclear RNA, and U(50) in 5 S rRNA. Pus7p belongs to the universally conserved TruD-like family of RNA:Psi-synthases found in bacteria, archaea, and eukarya. Although several RNA substrates for yeast Pus7p have been identified, specificity of their recognition and modification has not been studied. However, conservation of a 7-nt-long sequence, including the modified U residue, in all natural Pus7p substrates suggested the importance of these nucleotides for Pus7p recognition and/or catalysis. Using site-directed mutagenesis, we designed a set of RNA variants derived from the yeast tRNA(Asp)(GUC), pre-tRNA(Tyr)(GPsiA), and U2 small nuclear RNA and tested their ability to be modified by Pus7p in vitro. We demonstrated that the highly conserved U(-2) and A(+1) residues (nucleotide numbers refer to target U(0)) are crucial identity elements for efficient modification by Pus7p. Nucleotide substitutions at other surrounding positions (-4, -3, +2, +3) have only a moderate effect. Surprisingly, the identity of the nucleotide immediately 5' to the target U(0) residue (position -1) is not important for efficient modification. Alteration of tRNA three-dimensional structure had no detectable effect on Pus7p activity at position 13. However, our results suggest that the presence of at least one stem-loop structure including or close to the target U nucleotide is required for Pus7p-catalyzed modification.


Glucose-lactate metabolic cooperation in cancer: insights from a spatial mathematical model and implications for targeted therapy.
A recent study has hypothesised a glucose-lactate metabolic symbiosis between adjacent hypoxic and oxygenated regions of a developing tumour, and proposed a treatment strategy to target this symbiosis. However, in vivo experimental support remains inconclusive. Here we develop a minimal spatial mathematical model of glucose-lactate metabolism to examine, in principle, whether metabolic symbiosis is plausible in human tumours, and to assess the potential impact of inhibiting it. We find that symbiosis is a robust feature of our model system-although on the length scale at which oxygen supply is diffusion-limited, its occurrence requires very high cellular metabolic activity-and that necrosis in the tumour core is reduced in the presence of symbiosis. Upon simulating therapeutic inhibition of lactate uptake, we predict that targeted treatment increases the extent of tissue oxygenation without increasing core necrosis. The oxygenation effect is correlated strongly with the extent of wild-type hypoxia and only weakly with wild-type symbiotic behaviour, and therefore may be promising for radiosensitisation of hypoxic, lactate-consuming tumours even if they do not exhibit a spatially well-defined symbiosis. Finally, we conduct in vitro experiments on the U87 glioblastoma cell line to facilitate preliminary speculation as to where highly malignant tumours might fall in our parameter space, and find that these experiments suggest a weakly symbiotic regime for U87 cells, thus raising the new question of what relationship might exist between symbiosis and tumour malignancy.


Schwannoma development is mediated by Hippo pathway dysregulation and modified by RAS/MAPK signaling.
Schwannomas are tumors of the Schwann cells that cause chronic pain, numbness, and potentially life-threatening impairment of vital organs. Despite the identification of causative genes, including NF2 (Merlin), INI1/SMARCB1, and LZTR1, the exact molecular mechanism of schwannoma development is still poorly understood. Several studies have identified Merlin as a key regulator of the Hippo, MAPK, and PI3K signaling pathways; however, definitive evidence demonstrating the importance of these pathways in schwannoma pathogenesis is absent. Here, we provide direct genetic evidence that dysregulation of the Hippo pathway in the Schwann cell lineage causes development of multiple schwannomas in mice. We found that canonical Hippo signaling through the effectors YAP/TAZ is required for schwannomagenesis and that MAPK signaling modifies schwannoma formation. Furthermore, cotargeting YAP/TAZ transcriptional activity and MAPK signaling demonstrated a synergistic therapeutic effect on schwannomas. Our new model provides a tractable platform to dissect the molecular mechanisms underpinning schwannoma formation and the role of combinatorial targeted therapy in schwannoma treatment.


Cell damage in near-infrared multimode optical traps as a result of multiphoton absorption.
We report on cell damage of single cells confined in continuous-wave (cw), near-infrared (NIR) multimode optical traps as a result of multiphoton absorption phenomena. Trapping beams at NIR wavelengths less than 800 nm are capable of damaging cells through a two-photon absorption process. Cell damage is more pronounced in multimode cw traps compared with single-frequency true cw NIR traps because of transient power enhancement by longitudinal mode beating. Partial mode locking in tunable cw Ti:sapphire lasers used as trapping beam sources can produce unstable subnanosecond pulses at certain wavelengths that amplify multiphoton absorption effects significantly. We recommend the use of single-frequency long-wavelength NIR trapping beams for optical micromanipulation of vital cells.


A phase II trial of preoperative concurrent chemotherapy and dose escalated intensity modulated radiotherapy (IMRT) for locally advanced rectal cancer.
Objectives: To determine the pathological response rates and toxicity and in patients with locally advanced rectal cancer treated with concurrent capecitabine and dose escalated intensity modulated radiotherapy (IMRT) Methods: Patients with stage II or III adenocarcinoma of the rectum were treated with preoperative concurrent capecitabine and IMRT. Dose of capecitabine was 825mg/m2, 5 days a week for 5 weeks. IMRT was used to deliver a dose of 45Gy in 25 fractions (1.8Gy per fraction daily, 5 days a week over 5 weeks) to the regional lymphatics and areas at risk of harbouring microscopic disease. A concomitant synchronous integrated boost (SIB) to the gross tumour with a margin to a total dose of 55Gy in 25 fractions was also delivered in the same period. TME surgery was performed 8 weeks after preoperative therapy. The primary endpoint is pathological complete response rate (pCR) and the secondary endpoint was downstaging rates, Sphincter preservation rates (SPR), disease free survival (DFS) at 2 years and toxicity graded using the CTCAE v3.0. Results: Twenty three patients were enrolled. Three were not evaluable; one did not complete treatment due to logistic issues and two declined surgery. The remaining 20 patients completed preoperative chemoIMRT followed by TME surgery. At a median follow-up of 38.2 months (17.5-53.2 months), 90% (18 of 20) patients were alive. The 2 year overall survival and DFS were 90% and 90% respectively. 35%(7/20) of patients had a pCR. 65% (13 of 20) patients had successful downstaging of their rectal tumours. There was no local recurrence. Sphincter preservation rate was 85%. Treatment was well tolerated with only one patient (5%) having Grade 3 radiation proctitis. Conclusions: Preoperative concurrent capecitabine and dose escalated IMRT is well tolerated and results in high rates of pCR. A randomized trial comparing this regimen with standard 3D conformal chemoradiotherapy is warranted.


Cervical Radiculopathy: Effectiveness of Adding Traction to Physical Therapy-A Systematic Review and Meta-Analysis of Randomized Controlled Trials.
Cervical radiculopathy (CR) is a common cervical spine disorder. Cervical traction (CT) is a frequently recommended treatment for patients with CR. The purpose of this study was to conduct a review and meta-analysis of randomized controlled trials (RCTs) on the effect of CT combined with other physical therapy procedures versus physical therapy procedures alone on pain and disability. Data were obtained from COCHRANE Controlled Trials Register, PubMed, CINAHL, Scopus, ISI Web of Science, and PEDro, from their inception to July 2016. All RCTs on symptomatic adults with CR, without any restriction regarding publication time or language, were considered. Two reviewers selected the studies, conducted the quality assessment, and extracted the results. Meta-analysis employed a random-effects model. The evidence was assessed using GRADE criteria. Five studies met the inclusion criteria. Mechanical traction had a significant effect on pain at short- and intermediate-terms (g = -0.85 [95% CI = -1.63 to -0.06] and g = -1.17 [95% CI = -2.25 to -0.10], respectively) and significant effects on disability at intermediate term (g = -1.05; 95% CI = -1.81 to -0.28). Manual traction had significant effects on pain at short- term (g = -0.85; 95% CI = -1.39 to -0.30). The most important limitation of the present work is the lack of homogeneity in CR diagnostic criteria among the included studies. In light of these results, the current literature lends some support to the use of the mechanical and manual traction for CR in addition to other physical therapy procedures for pain reduction, but yielding lesser effects on function/disability.


Wide-spectral/dynamic-range skin-compatible phototransistors enabled by floated heterojunction structures with surface functionalized SWCNTs and amorphous oxide semiconductors.
Purified semiconducting single-walled carbon nanotubes (sc-SWCNTs) have been researched for optoelectronic applications due to their high absorption coefficient from the visible to even the near-infrared (NIR) region. Nevertheless, the insufficient electrical characteristics and incompatibility with conventional CMOS processing have limited their wide utilization in this emerging field. Here, we demonstrate highly detective and wide spectral/dynamic range phototransistors incorporating floated heterojunction active layers which are composed of low-temperature sol-gel processed n-type amorphous indium gallium zinc oxide (a-IGZO) stacked with a purified p-type sc-SWCNT layer. To achieve a high and broad spectral/dynamic range photo-response of the heterogeneous transistors, photochemically functionalized sc-SWCNT layers were carefully implemented onto the a-IGZO channel area with a floating p-n heterojunction active layer, resulting in the suppression of parasitic charge leakage and good bias driven opto-electrical properties. The highest photosensitivity (R) of 9.6 × 102 A W-1 and a photodetectivity (D*) of 4 × 1014 Jones along with a dynamic range of 100-180 dB were achieved for our phototransistor in the spectral range of 400-780 nm including continuous and minimal frequency independent behaviors. More importantly, to demonstrate the diverse application of the ultra-flexible hybrid photosensor platform as skin compatible electronics, the sc-SWCNT/a-IGZO phototransistors were fabricated on an ultra-thin (∼1 μm) polyimide film along with a severe static and dynamic electro-mechanical test. The skin-like phototransistors showed excellent mechanical stability such as sustainable good electrical performance and high photosensitivity in a wide dynamic range without any visible cracks or damage and little noise interference after being rolled-up on the 150 μm-thick optical fiber as well as more than 1000 times cycling.


Cells, Materials, and Fabrication Processes for Cardiac Tissue Engineering.
Cardiovascular disease is the number one killer worldwide, with myocardial infarction (MI) responsible for approximately 1 in 6 deaths. The lack of endogenous regenerative capacity, added to the deleterious remodelling programme set into motion by myocardial necrosis, turns MI into a progressively debilitating disease, which current pharmacological therapy cannot halt. The advent of Regenerative Therapies over 2 decades ago kick-started a whole new scientific field whose aim was to prevent or even reverse the pathological processes of MI. As a highly dynamic organ, the heart displays a tight association between 3D structure and function, with the non-cellular components, mainly the cardiac extracellular matrix (ECM), playing both fundamental active and passive roles. Tissue engineering aims to reproduce this tissue architecture and function in order to fabricate replicas able to mimic or even substitute damaged organs. Recent advances in cell reprogramming and refinement of methods for additive manufacturing have played a critical role in the development of clinically relevant engineered cardiovascular tissues. This review focuses on the generation of human cardiac tissues for therapy, paying special attention to human pluripotent stem cells and their derivatives. We provide a perspective on progress in regenerative medicine from the early stages of cell therapy to the present day, as well as an overview of cellular processes, materials and fabrication strategies currently under investigation. Finally, we summarise current clinical applications and reflect on the most urgent needs and gaps to be filled for efficient translation to the clinical arena.


Socioeconomic and physical distance to the maternity hospital as predictors for place of delivery: an observation study from Nepal.
BACKGROUND: Although the debate on the safety and women's right of choice to a home delivery vs. hospital delivery continues in the developed countries, an undesirable outcome of home delivery, such as high maternal and perinatal mortality, is documented in developing countries. The objective was to study whether socio-economic factors, distance to maternity hospital, ethnicity, type and size of family, obstetric history and antenatal care received in present pregnancy affected the choice between home and hospital delivery in a developing country. METHODS: This cross-sectional study was done during June, 2001 to January 2002 in an administratively and geographically well-defined territory with a population of 88,547, stretching from urban to adjacent rural part of Kathmandu and Dhading Districts of Nepal with maximum of 5 hrs of distance from Maternity hospital. There were no intermediate level of private or government hospital or maternity homes in the study area. Interviews were carried out on 308 women who delivered within 45 days of the date of the interview with a pre-tested structured questionnaire. RESULTS: A distance of more than one hour to the maternity hospital (OR = 7.9), low amenity score status (OR = 4.4), low education (OR = 2.9), multi-parity (OR = 2.4), and not seeking antenatal care in the present pregnancy (OR = 4.6) were statistically significantly associated with an increased risk of home delivery. Ethnicity, obstetric history, age of mother, ritual observance of menarche, type and size of family and who is head of household were not statistically significantly associated with the place of delivery. CONCLUSIONS: The socio-economic standing of the household was a stronger predictor of place of delivery compared to ethnicity, the internal family structure such as type and size of family, head of household, or observation of ritual days by the mother of an important event like menarche. The results suggested that mothers, who were in the low-socio-economic scale, delivered at home more frequently in a developing country like Nepal.


Uncompensated Care Decreased At Hospitals In Medicaid Expansion States But Not At Hospitals In Nonexpansion States.
One pillar of the Affordable Care Act (ACA) was its expected impact on the growing burden of uncompensated care costs for the uninsured at hospitals. However, little is known about how this burden changed as a result of the ACA's enactment. We examine how the Affordable Care Act (ACA)'s coverage expansions affected uncompensated care costs at a large, diverse sample of hospitals. We estimate that in states that expanded Medicaid under the ACA, uncompensated care costs decreased from 4.1 percentage points to 3.1 percentage points of operating costs. The reductions in Medicaid expansion states were larger at hospitals that had higher pre-ACA uncompensated care burdens and in markets where we predicted larger gains in coverage through expanded eligibility for Medicaid. Our estimates suggest that uncompensated care costs would have decreased from 5.7 percentage points to 4.0 percentage points of operating costs in nonexpansion states if they had expanded Medicaid. Thus, while the ACA decreased the variation in uncompensated care costs across hospitals within Medicaid expansion states, the difference between expansion and nonexpansion states increased substantially. Policy makers and researchers should consider how the shifting uncompensated care burden affects other hospital decisions as well as the distribution of supplemental public funding to hospitals.


Arginine vasopressin versus norepinephrine: will the stronger one win the race?
In the current issue of Critical Care, Friesenecker and colleagues present a well-designed comparative study on the microvascular effects of arginine vasopressin (AVP) and norepinephrine (NE) in a physiological, unanesthetized hamster model. The authors clearly demonstrate that AVP, but not NE, has marked vasoconstrictive effects on large arterioles, whereas the impact on small arterioles is comparable for both vasopressors. However, it remains unclear if these results, per se, reflect a stronger vasopressive potential of AVP versus NE, as macrohemodynamic variables were not different between study groups. Since the authors did not investigate the effects of AVP and NE in vasodilatory shock states, the microcirculatory response in sepsis or systemic inflammatory response syndrome remains inconclusive. The same authors previously reported that AVP infusion in patients suffering from vasodilatory shock carries the risk for ischemic skin lesions. This in turn raises the question whether the quality of vasopressors should be judged by their potency.


The best and worst aspects of the ICU research coordinator role.
The Research Coordinator (RC) role is a relative new addition to staffing profiles in Australasian Intensive Care Units (ICUs). The RC plays a pivotal role in conducting ethically and scientifically sound research. There have been anecdotal reports of the RC role in various speciality areas. However, limited research has been undertaken into the role, and only one study has been published from the intensive care setting. To evaluate which factors the RC found to be the best and worst attributes of the role. A cross-sectional web-based cohort study was conducted in 2004 collecting free text information from RCs in ICUs in Australia and New Zealand. Forty-nine participants (71%) completed the study with 273 entries into the two categories of 'best' (60%) and 'worst' (40%) aspects of the role. We identified four thematic clusters in both categories: (1) 'How the job was structured' (2) 'The worth of the job' (3) 'What the work involves' and (4) 'Who I work with'. Both categories received proportionally the same number of responses in each theme with (1) the most frequent and (4) the least number of entries. There is much variation between RCs in almost every descriptor of the role. The Australasian Intensive Care RC values autonomy, respect and intellectual stimulation with the scope for extending the role to suit the individual and finds isolation, under-recognition and workload, often not sufficiently compensated, to be the worst aspects of the position.


Semi-Supervised Pipeline for Autonomous Annotation of SARS-CoV-2 Genomes.
SARS-CoV-2 genomic sequencing efforts have scaled dramatically to address the current global pandemic and aid public health. However, autonomous genome annotation of SARS-CoV-2 genes, proteins, and domains is not readily accomplished by existing methods and results in missing or incorrect sequences. To overcome this limitation, we developed a novel semi-supervised pipeline for automated gene, protein, and functional domain annotation of SARS-CoV-2 genomes that differentiates itself by not relying on the use of a single reference genome and by overcoming atypical genomic traits that challenge traditional bioinformatic methods. We analyzed an initial corpus of 66,000 SARS-CoV-2 genome sequences collected from labs across the world using our method and identified the comprehensive set of known proteins with 98.5% set membership accuracy and 99.1% accuracy in length prediction, compared to proteome references, including Replicase polyprotein 1ab (with its transcriptional slippage site). Compared to other published tools, such as Prokka (base) and VAPiD, we yielded a 6.4- and 1.8-fold increase in protein annotations. Our method generated 13,000,000 gene, protein, and domain sequences-some conserved across time and geography and others representing emerging variants. We observed 3362 non-redundant sequences per protein on average within this corpus and described key D614G and N501Y variants spatiotemporally in the initial genome corpus. For spike glycoprotein domains, we achieved greater than 97.9% sequence identity to references and characterized receptor binding domain variants. We further demonstrated the robustness and extensibility of our method on an additional 4000 variant diverse genomes containing all named variants of concern and interest as of August 2021. In this cohort, we successfully identified all keystone spike glycoprotein mutations in our predicted protein sequences with greater than 99% accuracy as well as demonstrating high accuracy of the protein and domain annotations. This work comprehensively presents the molecular targets to refine biomedical interventions for SARS-CoV-2 with a scalable, high-accuracy method to analyze newly sequenced infections as they arise.


Stress as a potential modifier of the impact of lead levels on blood pressure: the normative aging study.
Lead exposure and psychological stress have been independently associated with hypertension in various populations, and animal studies suggest that when they co-occur, their effects may be exacerbated. We examined whether psychological stress modifies the impact of cumulative lead exposure (measured as bone lead levels) on hypertension and blood pressure in Boston-area community-exposed men participating in the Normative Aging Study. We evaluated the modifying effect of stress on lead exposure on baseline hypertension status (513 participants) and on blood pressure in those without hypertension (237 participants), cross-sectionally. In baseline nonhypertensives, we examined the same risk factors in relation to prospective risk of developing hypertension. Cross-sectional analysis revealed a positive interaction between stress and tibia lead on systolic blood pressure, after adjusting for age, body mass index, family history of high blood pressure, education, smoking, alcohol consumption, physical activity, and nutritional factors. In prospective multivariate analyses, high stress also modified the effect of tibia lead and patella lead on the risk of developing hypertension. Those reporting high stress had 2.66 [95% confidence interval (CI), 1.43-4.95] times the risk of developing hypertension per standard deviation increase in tibia lead and had 2.64 (95% CI, 1.42-4.92) times the risk per standard deviation increase in patella lead. To our knowledge, these are the first analyses to look at interactive effects of stress and lead on hypertension in humans. These results suggest that the effect of lead on hypertension is most pronounced among highly stressed individuals, independent of demographic and behavioral risk factors.


Validity of clinical measures of shoulder subluxation in adults with poststroke hemiplegia.
Shoulder subluxation after cerebrovascular accident has been associated with chronic pain, orthopedic complications, peripheral nerve damage, and autonomic dysfunction. Clinical diagnosis and gradation of subluxation is problematic due to the lack of precision in frequently used measures. The objective of this study was to assess the validity of clinical techniques used to assess inferior subluxation of the hemiplegic shoulder by comparing these techniques with radiographic measurement. In 20 male subjects with hemiplegia, the presence, type, and degree of subluxation was assessed with three clinical measures: palpation, arm length discrepancy, and thermoplastic jig measurement. Anterior-posterior X rays of the hemiplegic shoulder were taken after clinical examination. Spearman rank correlation coefficients between the X rays and the three clinical measures were relatively low. Palpation had the highest correlation (rs = .76), followed by arm length discrepancy (rs = .46), and thermoplastic jig measurement (rs = .42). These findings provide cautious optimism about using these clinical measures to identify subluxation. Although detection was best with palpation, the likely inability to determine clinical overcorrecting of subluxation makes use of palpation alone suspect. Improved techniques of arm length measurement may provide a solution to this problem. These findings further necessitate that improved procedures for clinical assessment of subluxation be developed.