Possibility of Immediate Introduction of a Single-Dose Antibody Induction Test as a Refinement of the NIH Test for Inactivated Rabies Vaccine Potency Determination.
Antibody induction test (AIT) is a promising candidate as a refinement of the troublesome National institutes of Health (NIH) test in the sense of animal welfare 3R approach for determination of potency of inactivated rabies vaccines for veterinary and human use. In this study, we initially try to develop AIT as a suitable alternative to NIH test, to achieve a reduction of test duration and diminish animal suffering by omitting intracerebral CVS infection and measuring humoral immunity upon vaccination. Designs of both multi-dose and single-dose AIT were examined. Biological reference preparation, batch 5 with assigned titer of 10 IU/vial, was taken as both standard and test vaccine. Six consecutive AITs were performed and eight pools of sera in each AIT were tested in triplicate by rapid fluorescent focus inhibition test. We estimated the upper detection limit and calculated test variability for individual dilutions. For multi-dose AIT, we estimated the dose-response function and performed calculations of final test results and statistical validity parameters for both linear and sigmoidal model using CombiStats program. Sigmoidal 4-parameter dose-response model was found optimal. Presented design of multi-dose AIT showed a satisfactory detection limit for testing of inactivated rabies vaccines for both veterinary and human use. However, due to nonconformity of obtained results with statistical validity criteria, we concluded that the presented model of multi-dose AIT was unsuitable for introduction in routine practice. However, we concluded that there was a realistic option for introduction of two versions of single-dose AIT. The first version would be with two standard vaccine controls and could be introduced immediately, while the second version would include testing of the sample only and rely on comparison of the induced rabies antibody level with absolute cut-off limits set in advance.


Gap junctional conductance in ventricular myocyte pairs isolated from postischemic rabbit myocardium.
Abnormalities of myocardial gap junction-mediated cell coupling have been implicated in cardiac arrhythmogenesis. The potential role of gap junctional dysfunction in the generation of reperfusion-induced arrhythmias is uncertain. The purpose of this study was to measure the effects of myocardial ischemia and reperfusion on gap junctional conductance (gj) between isolated ventricular myocytes. By using a new experimental model, myocyte pairs were isolated from Langendorff-perfused rabbit hearts 1) after 30 minutes of global normothermic ischemia followed by 30 minutes of reperfusion, 2) after 75 minutes of control perfusion, or 3) immediately after removal of the heart. Myocytes and myocyte pairs were studied using whole-cell recording techniques. Action potential characteristics of cells in all three groups were normal. Despite similar mean gj in all three groups (0.88 +/- 0.27, 1.15 +/- 0.18, and 1.24 +/- 0.25 microS, respectively; p greater than 0.05), the postischemic group was more widely distributed and had a significantly greater proportion of poorly communicating cell pairs than either control group (gj less than 25% of mean in eight of 15 myocyte pairs versus zero of 15 and one of 13, respectively; p less than 0.02). Thus, postischemic myocyte pairs represent a heterogeneous population of electrically coupled cells in which individual deficits in coupling are masked by a normal mean value. In the reperfused intact heart, local disturbances of cell coupling, similarly undetected by gross measures of conduction, could disrupt myocardial conduction and activation on a microscopic scale and thus enhance arrhythmogenicity.


Modulation of cyclin dependent kinase inhibitor proteins and ERK1/2 activity in allylamine-injured vascular smooth muscle cells.
Chronic oxidative injury by allylamine (AAM) induces proliferative vascular smooth muscle cell (vSMC) phenotypes in the rat aorta similar to those seen in rodent and human atherosclerotic lesions. The proliferative advantage of AAM vSMC compared to control cells is maintained with serial passage of the cells and the advantage is nullified when AAM cells are seeded on a collagen substrate. In this study, we evaluate the potential role of cyclin dependent kinase inhibitors, p27 and p21, and mitogen activated protein (MAP) kinases, ERK1/2, in mediating the proliferative advantage of AAM stressed vSMC over control cells on plastic or collagen substrates. p27 levels in randomly cycling cells were comparable in both cell types irrespective of the substrate. In contrast, basal levels of p21 were 1.9 +/- 0.3 (P < 0.05)-fold higher in randomly cycling AAM cells seeded on plastic compared to controls, a difference that was lost on a collagen substrate. Following G0 synchronization, basal levels of both p27 and p21 were higher in AAM cells seeded on plastic compared to controls (1.7 +/- 0.2 and 2.0 +/- 0.3-fold, respectively, P < 0.05), but these differences were lost upon mitogenic stimulation. Pyrrolidine dithiocarbamate (PDTC) decreased p27 and p21 levels in cycling AAM cells relative to controls in a substrate-dependent manner. AAM cells seeded on plastic exhibited enhanced ERK1/2 activation upon mitogenic stimulation; seeding on collagen nullified this advantage. The duration of ERK1/2 activation was prolonged in AAM cells independently of the seeding substrate. We conclude that substrate-dependent acquisition of proliferative phenotypes following repeated cycles of AAM injury correlates with modulation of the cyclin dependent kinase inhibitors, p27 and p21.


Pre- and Postnatal Neuroimaging of Congenital Cerebellar Abnormalities.
The human cerebellum has a protracted development that makes it vulnerable to a broad spectrum of developmental disorders including malformations and disruptions. Starting from 19 to 20 weeks of gestation, prenatal magnetic resonance imaging (MRI) can reliably study the developing cerebellum. Pre- and postnatal neuroimaging plays a key role in the diagnostic work-up of congenital cerebellar abnormalities. Diagnostic criteria for cerebellar malformations and disruptions are based mostly on neuroimaging findings. The diagnosis of a Dandy-Walker malformation is based on the presence of hypoplasia, elevation, and counterclockwise upward rotation of the cerebellar vermis and cystic dilatation of the fourth ventricle, which extends posteriorly filling out the posterior fossa. For the diagnosis of Joubert syndrome, the presence of the molar tooth sign (thickened, elongated, and horizontally orientated superior cerebellar peduncles and an abnormally deep interpeduncular fossa) is needed. The diagnostic criteria of rhombencephalosynapsis include a complete or partial absence of the cerebellar vermis and continuity of the cerebellar hemispheres across the midline. Unilateral cerebellar hypoplasia is defined by the complete aplasia or hypoplasia of one cerebellar hemisphere. Familiarity with these diagnostic criteria as well as the broad spectrum of additional neuroimaging findings is important for a correct pre- and postnatal diagnosis. A correct diagnosis is essential for management, prognosis, and counseling of the affected children and their family.


Prevalence and Frequency of mHealth and eHealth Use Among US and UK Smokers and Differences by Motivation to Quit.
Both mHealth and eHealth interventions for smoking cessation are rapidly being developed and tested. There are no data on use of mHealth and eHealth technologies by smokers in general or by smokers who are not motivated to quit smoking. The aims of our study were to (1) assess technology use (eg, texting, social media, Internet) among smokers in the United States and United Kingdom, (2) examine whether technology use differs between smokers who are motivated to quit and smokers who are not motivated to quit, (3) examine previous use of technology to assist with smoking cessation, and (4) examine future intentions to use technology to assist with smoking cessation. Participants were 1000 adult smokers (54.90%, 549/1000 female; mean age 43.9, SD 15.5 years; US: n=500, UK: n=500) who were recruited via online representative sampling strategies. Data were collected online and included demographics, smoking history, and frequency and patterns of technology use. Among smokers in general, there was a high prevalence of mobile and smartphone ownership, sending and receiving texts, downloading and using apps, using Facebook, and visiting health-related websites. Smokers who were unmotivated to quit were significantly less likely to own a smartphone or handheld device that connects to the Internet than smokers motivated to quit. There was a significantly lower prevalence of sending text messages among US smokers unmotivated to quit (78.2%, 179/229) versus smokers motivated to quit (95.0%, 229/241), but no significant differences between the UK groups (motivated: 96.4%, 239/248; unmotivated: 94.9%, 223/235). Smokers unmotivated to quit in both countries were significantly less likely to use a handheld device to read email, play games, browse the Web, or visit health-related websites versus smokers motivated to quit. US smokers had a high prevalence of app downloads regardless of motivation to quit, but UK smokers who were motivated to quit had greater prevalence of app downloads than smokers unmotivated to quit. US smokers were significantly more likely to have a Facebook account (87.0%, 435/500) than UK smokers (76.4%, 382/500), but smokers unmotivated to quit in both countries used Facebook less frequently than smokers motivated to quit. Smokers who were unmotivated to quit were less likely to have used eHealth or mHealth platforms to help them quit smoking in the past and less likely to say that they would use them for smoking cessation in the future. Although smokers unmotivated to quit make less use of technology than smokers motivated to quit, there is sufficient prevalence to make it worthwhile to develop eHealth and mHealth interventions to encourage cessation. Short and low-effort communications, such as text messaging, might be better for smokers who are less motivated to quit. Multiple channels may be required to reach unmotivated smokers.


Peristaltic transport of a power-law fluid with variable consistency.
Effects of the consistency variation on the peristaltic transport of a non-Newtonian power-law fluid through a tube have been investigated by taking into account the existence of a peripheral layer. It is shown that the flow rate flux, for zero pressure drop, increases as the amplitude of the peristaltic wave increases but it decreases due to the pseudoplastic nature of the fluid. It is also noted that, for zero pressure drop, the flux does not depend on the consistency of peripheral layer while the friction decreases as this consistency decreases. However, for nonzero pressure drop, the flux increases and the friction force decreases as the consistency of peripheral layer fluid decreases.


Observations of the seasonal buildup and washout of salts in urban bioswale soil.
The objective of this study was to quantify the seasonal risk of salt damage to bioswale plants, soil, microbes, and downstream waterbodies. To do so, we measured sodium, chloride, and electrical conductivity levels at seven bioswales located in the Bronx, New York City, over 42 storm events during a three-year monitoring period. The bioswale with the greatest salt contamination (median 206 mg/L chloride) had a unique inlet design without any possibility of inlet bypass. The most severe effects at all sites were found during the winter season, as infiltrate concentrations frequently (40% of winter samples) exceeded 1000 mg/L chloride, a level lethal to aquatic plants and invertebrates, and electrical conductivity exceeded 1500 μS cm-1 (50% of winter samples), a level that may displace bound metals from bioswale soils and into the subsurface. However, low levels of permanent salt contamination may be expected all year, as concentrations frequently (87% of all samples) exceeded the United States Environmental Protection Agency drinking water standard of 20 mg/L sodium. A regression of chloride washout over the year yielded concentrations greater than those damaging to soil structure and soil microbes (90 mg/L) until August 20th, and above those damaging to roadside vegetation (30 mg/L) for the entire year. Today, the vast majority of bioswales in cold climates are built with salt-tolerant vegetation, but prior to this study, it was unclear to what degree this was, in fact, necessary. Our findings confirm salt-tolerant vegetation to be optimal, as winter de-icing salts are not sufficiently flushed from soils by the spring growing season. Our findings also demonstrate how bioswale inlet design and site location can influence soil contamination.


Visual claustrum: topography and receptive field properties in the cat.
A region containing visually responsive cells was found in the dorsocaudal claustrum. This area contains a single orderly map of the contralateral visual field. Like cortical cells, most claustral cells are selective for stimulus orientation. They are binocular, and they respond to either direction of movement and to a broad range of velocities. Their most striking property is a marked preference for very long stimuli.


Reverse transcriptase activity and Ty RNA are associated with virus-like particles in yeast.
The Ty element of yeast represents a class of eukaryotic transposons that show remarkable structural similarity to retroviral proviruses. Recently, these comparisons have been strengthened by a series of observations on the yeast Ty element: Ty transposes via an RNA intermediate; it contains a sequence (Fig. 1) which, when translated, is homologous to a conserved region found in all reverse transcriptases; a fusion protein encoded by Ty is produced by a frameshift event that is directly analogous to the production of Pr180gag-pol in a retrovirus such as Rous sarcoma virus. Here we identify the reverse transcriptase activity that, until now, has been presumed to mediate Ty transposition and show that it is sequestered in virus-like particles that also contain Ty RNA.


Active pain coping is associated with the response in real-time fMRI neurofeedback during pain.
Real-time functional magnetic resonance imaging (rt-fMRI) neurofeedback is used as a tool to gain voluntary control of activity in various brain regions. Little emphasis has been put on the influence of cognitive and personality traits on neurofeedback efficacy and baseline activity. Here, we assessed the effect of individual pain coping on rt-fMRI neurofeedback during heat-induced pain. Twenty-eight healthy subjects completed the Coping Strategies Questionnaire (CSQ) prior to scanning. The first part of the fMRI experiment identified target regions using painful heat stimulation. Then, subjects were asked to down-regulate the pain target brain region during four neurofeedback runs with painful heat stimulation. Functional MRI analysis included correlation analysis between fMRI activation and pain ratings as well as CSQ ratings. At the behavioral level, the active pain coping (first principal component of CSQ) was correlated with pain ratings during neurofeedback. Concerning neuroimaging, pain sensitive regions were negatively correlated with pain coping. During neurofeedback, the pain coping was positively correlated with activation in the anterior cingulate cortex, prefrontal cortex, hippocampus and visual cortex. Thermode temperature was negatively correlated with anterior insula and dorsolateral prefrontal cortex activation. In conclusion, self-reported pain coping mechanisms and pain sensitivity are a source of variance during rt-fMRI neurofeedback possibly explaining variations in regulation success. In particular, active coping seems to be associated with successful pain regulation.


HyBeacon probes for rapid DNA sequence detection and allele discrimination.
HyBeacon probes are single-stranded oligonucleotides with one or more internal base(s) labeled with a fluorescent dye. When a probe forms a duplex with its target sequence, the level of fluorescence emission increases considerably. HyBeacons have been developed as new tools for rapid sequence detection and discrimination and have been employed in a wide variety of applications including infectious diagnostics and analysis of human polymorphisms. Single-labeled (FVG1) and dual-labeled (FVG11) probes were designed to analyze the factor V Leiden (R506Q) polymorphism which causes an increased risk of deep vein thrombosis and pulmonary embolism. Detection and identification of factor V alleles is performed by melting curve analysis and determination of probe melting temperature (T(m)). HyBeacon hybridization to the glutamine allele (Q) causes the formation of mismatched DNA duplexes that are detected through decreases in T(m). HyBeacon probes are included in homogeneous PCR assays to genotype samples with respect to the factor V polymorphism within 20 min, using purified DNAs and unpurified saliva/blood samples. This paper describes the preparation of homogeneous PCR assays, LightCycler target amplification, and subsequent melting curve analysis. This chapter also describes the use of homologous oligonucleotides and melting curve analysis as a method for probe evaluation.


Enforced telomere elongation increases the sensitivity of human tumour cells to ionizing radiation.
More than 85% of all human cancers possess the ability to maintain chromosome ends, or telomeres, by virtue of telomerase activity. Loss of functional telomeres is incompatible with survival, and telomerase inhibition has been established in several model systems to be a tractable target for cancer therapy. As human tumour cells typically maintain short equilibrium telomere lengths, we wondered if enforced telomere elongation would positively or negatively impact cell survival. We found that telomere elongation beyond a certain length significantly decreased cell clonogenic survival after gamma irradiation. Susceptibility to irradiation was dosage-dependent and increased at telomere lengths exceeding 17kbp despite the fact that all chromosome ends retained telomeric DNA. These data suggest that an optimal telomere length may promote human cancer cell survival in the presence of genotoxic stress.


Is there a kink in consumers' threshold value for cost-effectiveness in health care?
A reproducible observation is that consumers' willingness-to-accept (WTA) monetary compensation to forgo a program is greater than their stated willingness-to-pay (WTP) for the same benefit. Several explanations exist, including the psychological principle that the utility of losses weighs heavier than gains. We sought to quantify the WTP-WTA disparity from published literature and explore implications for cost-effectiveness analysis accept-reject thresholds in the south-west quadrant of the cost-effectiveness plane (less effect, less cost). We reviewed published studies (health and non-health) to estimate the ratio of WTA to WTP for the same program benefit for each study and to determine if WTA is consistently greater than WTP in the literature. WTA/WTP ratios were greater than unity for every study we reviewed. The ratios ranged from 3.2 to 89.4 for environmental studies (n=7), 1.9 to 6.4 for health care studies (n=2), 1.1 to 3.6 for safety studies (n=4) and 1.3 to 2.6 for experimental studies (n=7). Given that WTA is greater than WTP based on individual preferences, should not societal preferences used to determine cost-effectiveness thresholds reflect this disparity? Current convention in cost-effectiveness analysis is that any given accept-rejection criterion (e.g. $50 k/QALY gained) is symmetric - a straight line through the origin of the cost-effectiveness plane. The WTA-WTP evidence suggests a downward 'kink' through the origin for the south-west quadrant, such that the 'selling price' of a QALY is greater than the 'buying price'. The possibility of 'kinky cost-effectiveness' decision rules and the size of the kink merits further exploration.


The unique contributions of perceiver and target characteristics in person perception.
Models of person perception have long asserted that our impressions of others are guided by characteristics of both the target and perceiver. However, research has not yet quantified to what extent perceivers and targets contribute to different impressions. This quantification is theoretically critical, as it addresses how much an impression arises from "our minds" versus "others' faces." Here, we apply cross-classified random effects models to address this fundamental question in social cognition, using approximately 700,000 ratings of faces. With this approach, we demonstrate that (a) different trait impressions have unique causal processes, meaning that some impressions are largely informed by perceiver-level characteristics whereas others are driven more by physical target-level characteristics; (b) modeling of perceiver- and target-variance in impressions informs fundamental models of social perception; (c) Perceiver × Target interactions explain a substantial portion of variance in impressions; (d) greater emotional intensity in stimuli decreases the influence of the perceiver; and (e) more variable, naturalistic stimuli increases variation across perceivers. Important overarching patterns emerged. Broadly, traits and dimensions representing inferences of character (e.g., dominance) are driven more by perceiver characteristics than those representing appearance-based appraisals (e.g., youthful-attractiveness). Moreover, inferences made of more ambiguous traits (e.g., creative) or displays (e.g., faces with less extreme emotions, less-controlled stimuli) are similarly driven more by perceiver than target characteristics. Together, results highlight the large role that perceiver and target variability play in trait impressions, and develop a new topography of trait impressions that considers the source of the impression. (PsycINFO Database Record


Free radical scavenging and antioxidative properties of 'silibin' complexes on microsomal lipid peroxidation.
The antioxidant properties of silibin complexes, the water-soluble form silibin dihemisuccinate (SDH), and the lipid-soluble form, silibin phosphatidylcholine complex known as IdB 1016, were evaluated by studying their abilities to react with the superoxide radical anion (O2-.), and the hydroxyl radical (OH.). In addition, their effect on pulmonary and hepatic microsomal lipid peroxidation had been investigated. Superoxide radicals were generated by the PMS-NADH system and measured by their ability to reduce NBT. IC50 concentrations for the inhibition of the NBT reduction by SDH and IdB 1016 were found to be 25 microM and 316 microM respectively. Both silibin complexes had an inhibitory effect on xanthine oxidase activity. SDH reacted rapidly with OH radicals at approximately diffusion controlled rate and the rate constant was found to be (K = 8.2 x 10(9) M-1 s-1); it appeared to chelate Fe2+ in solution. In hepatic microsomes, when lipid peroxidation was induced by Fe2+, SDH inhibited by 39.5 per cent and IdB 1016 by 19.5 per cent, whereas when lipid peroxidation was induced by CuOOH, IdB 1016 exerted a better protective effect than SDH (29.4 per cent and 19.4 per cent inhibition, respectively). In both microsomal systems lipid peroxidation proceeded through a thiol depletion mechanism which could be restored in the presence of silibin complexes. Low levels of lipid peroxidation in pulmonary microsomes point out the differences between in-vitro lipid peroxidation occurring in microsomes of different tissues. The results support the free radical scavenger and antioxidative properties of silibin when it is complexed with a suitable molecule to increase its bioavailability.


The Pitfalls of Febrile Jaundice. A Case Report.
Jaundice in sepsis is usually caused by cholestasis, and its onset can precede other manifestations of the infection. Inflammation-induced cholestasis is a common complication in patients with an extrahepatic infection or those with inflammatory processes. We describe the case of a 47 years old female who presented with low back pain and paravertebral muscular contracture. She subsequently developed a cholestatic syndrome with clinical manifestations such as jaundice, followed by fever and sepsis with multiple organ dysfunction. Initially labeled as biliary sepsis, the diagnosis was crucially reoriented as the blood cultures were positive for Streptococcus pyogenes and the magnetic resonance imaging (MRI) findings suggested spondylodiscitis as well as a paravertebral abscess.


Hyperventilation-induced cerebral ischemia in patients with acute brain lesions: demonstration by xenon-enhanced CT.
To examine the possibility that hyperventilation, commonly used to prevent or treat increased intracranial pressure in patients with acute brain lesions, may induce significant cerebral ischemia. Local cerebral blood flow and vascular reactivity were measured before and after hyperventilation using xenon-enhanced CT in 12 patients with acute brain lesions. Five patients showed "paradoxical" reactivity (increased cerebral blood flow during hyperventilation) within the lesions. In five patients, hyperventilation induced ischemia in apparently normal regions of brain. In three patients, areas of luxury perfusion became ischemic during hyperventilation, while in three patients, lesions with moderate ischemia became more ischemic. Most patients showed more than one type of reactivity. These findings document hyperventilation-induced ischemia in acute brain lesions, and demonstrate that this phenomenon affects both injured and apparently intact areas of the brain. Further studies are required to determine the clinical significance of these pathophysiologic changes.


Silicosis Among Immigrant Engineered Stone (Quartz) Countertop Fabrication Workers in California.
Silicosis associated with inhalation of respirable crystalline silica among engineered stone countertop fabrication workers is an emerging health concern. To describe clinical, socioeconomic, and occupational characteristics of patients diagnosed with silicosis associated with engineered stone in California. This case series included reported cases of silicosis associated with fabrication of engineered stone countertops, as identified by statewide surveillance by the California Department of Public Health (2019-2022). Data analysis was performed from October 2022 to March 2023. Patient interviews and medical record abstractions were used to assess occupational exposure to respirable crystalline silica, including duration of work tenure and preventive measures undertaken. Demographics, clinical characteristics, health care utilization, and clinical outcomes were obtained, including vital status, hypoxia, and lung transplant. This case series identified 52 male patients meeting inclusion criteria; median (IQR) age was 45 (40-49) years, and 51 were Latino immigrants. Ten (19%) were uninsured, and 20 (39%) had restricted-scope Medi-Cal; 25 (48%) presented initially to an emergency department. A delay in diagnosis occurred in 30 (58%) patients, most commonly due to alternative initial diagnoses of bacterial pneumonia (9 [30%]) or tuberculosis (8 [27%]). At diagnosis, 20 (38%) patients had advanced disease (progressive massive fibrosis) with severely or very severely reduced forced expiratory volume in 1 second in 8 (18%) and 5 (11%), respectively. Of the cases, 10 (19%) were fatal; median (IQR) age at death was 46 (38-51) years, and 6 patients (12%) were alive with chronic resting hypoxia. Eleven were referred for lung transplant: 3 underwent transplant with 1 fatality; 7 were declined transplant, with 6 fatalities; and 1 died prior to listing. Median (IQR) work tenure was 15 (10-20) years; 23 (45%) reported use of water suppression for dust mitigation, and 25 (48%) continued to fabricate stone after being diagnosed with silicosis. In this case series performed in California, silicosis associated with occupational exposure to dust from engineered stone primarily occurred among young Latino immigrant men. Many patients presented with severe disease, and some cases were fatal.


Liposomal glucocorticoids as tumor-targeted anti-angiogenic nanomedicine in B16 melanoma-bearing mice.
This study evaluates whether the inhibitory effects of prednisolone phosphate (PLP) encapsulated in long-circulating liposomes (LCL-PLP) on tumor growth and tumor angiogenesis described previously can be generalized to other types of glucocorticoids (GC) encapsulated in LCL (LCL-GC). Four types of synthetic GC, i.e. budesonide disodium phosphate (BUP), dexamethasone disodium phosphate (DXP), methylprednisolone disodium phosphate (MPLP), and PLP, were selected based on the difference in their potency to activate the human glucocorticoid receptor. The effects of all LCL-GC on the production of angiogenic/inflammatory factors in vivo in the B16.F10 murine melanoma model as well as on the viability and proliferation of tumor cells and endothelial cells in vitro were investigated. Our results show that all four selected LCL-GC formulations inhibit tumor growth, albeit to different degrees. The differences in antitumor activity of LCL-GC correlate with their efficacy to suppress tumor angiogenesis and inflammation. The strongest antitumor effect is achieved by LCL-encapsulated BUP (LCL-BUP), due to the highest potency of BUP versus the other three GC types. The in vitro results presented herein suggest that LCL-BUP has strong cytotoxic effects on B16.F10 melanoma cells and the anti-proliferative effects of all LCL-GC towards angiogenic endothelial cells play a role in their antitumor activity.


Surface plasmon-assisted microscope.
Total internal reflection microscopy (TIRF) has been a powerful tool in biological research. The most valuable feature of the method has been the ability to image 100- to 200-nm-thick layer of cell features adjacent to a coverslip, such as membrane lipids, membrane receptors, and structures proximal-to-basal membranes. Here, we demonstrate an alternative method of imaging thin-layer proximal-to-basal membranes by placing a sample on a high refractive index coverslip covered by a thin layer of gold. The sample is illuminated using the Kretschmann method (i.e., from the top to an aqueous medium). Fluorophores that are close to the metal surface induce surface plasmons in the metal film. Fluorescence from fluorophores near the metal surface couple with surface plasmons allowing them to penetrate the metal surface and emerge at a surface plasmon coupled emission angle. The thickness of the detection layer is further reduced in comparison with TIRF by metal quenching of fluorophores at a close proximity (below 10 nm) to a surface. Fluorescence is collected by a high NA objective and imaged by EMCCD or converted to a signal by avalanche photodiode fed by a single-mode optical fiber inserted in the conjugate image plane of the objective. The system avoids complications of through-the-objective TIRF associated with shared excitation and emission light path, has thin collection thickness, produces excellent background rejection, and is an effective method to study molecular motion.


Quantitation of acute experimental ocular inflammation with 111indium-leukocytes.
The cellular component of an acute ocular inflammation in rabbits was measured with autologous leukocytes exogenously labeled with 111Indium tropolonate. Inflammation was induced by intravitreal bacterial lipopolysaccharide (LPS). After 16 hr blood was removed, leukocytes separated, labeled with 111Indium tropolonate and reinjected. Three cell fractions were examined: a leukocyte rich fraction which had been prepared with Dextran; and polymorphonuclear and mononuclear leukocyte fractions which had been prepared using a discontinuous Percoll gradient. Two hours after labeled leukocytes were injected, measurements of 111Indium were made in blood, plasma, the whole eye and in ocular compartments. From these data the numbers of each leukocyte population present were estimated and compared directly to histopathologic changes. Both polymorphonuclear and mononuclear leukocytes entered ocular tissues during the 2 hr period beginning 20 hr after LPS injection. Altered ocular vascular permeability was successfully measured with 125Iodine-albumin in some of these same rabbits. Both the number and type of inflammatory cell entering ocular tissues during a set period of time of the inflammatory response could thus be measured. This technique provides an opportunity to define the relationship of leukocyte infiltration and altered ocular vascular permeability in ocular tissues during the inflammatory response.


The SeqAn C++ template library for efficient sequence analysis: A resource for programmers.
The use of novel algorithmic techniques is pivotal to many important problems in life science. For example the sequencing of the human genome (Venter et al., 2001) would not have been possible without advanced assembly algorithms and the development of practical BWT based read mappers have been instrumental for NGS analysis. However, owing to the high speed of technological progress and the urgent need for bioinformatics tools, there was a widening gap between state-of-the-art algorithmic techniques and the actual algorithmic components of tools that are in widespread use. We previously addressed this by introducing the SeqAn library of efficient data types and algorithms in 2008 (Döring et al., 2008). The SeqAn library has matured considerably since its first publication 9 years ago. In this article we review its status as an established resource for programmers in the field of sequence analysis and its contributions to many analysis tools. We anticipate that SeqAn will continue to be a valuable resource, especially since it started to actively support various hardware acceleration techniques in a systematic manner.


Age-related neural changes during memory conjunction errors.
Human behavioral studies demonstrate that healthy aging is often accompanied by increases in memory distortions or errors. Here we used event-related fMRI to examine the neural basis of age-related memory distortions. We used the memory conjunction error paradigm, a laboratory procedure known to elicit high levels of memory errors. For older adults, right parahippocampal gyrus showed significantly greater activity during false than during accurate retrieval. We observed no regions in which activity was greater during false than during accurate retrieval for young adults. Young adults, however, showed significantly greater activity than old adults during accurate retrieval in right hippocampus. By contrast, older adults demonstrated greater activity than young adults during accurate retrieval in right inferior and middle prefrontal cortex. These data are consistent with the notion that age-related memory conjunction errors arise from dysfunction of hippocampal system mechanisms, rather than impairments in frontally mediated monitoring processes.


Pseudomonas chloritidismutans sp. nov., a non-denitrifying, chlorate-reducing bacterium.
A Gram-negative, facultatively anaerobic, rod-shaped, dissimilatory chlorate-reducing bacterium, strain AW-1(T), was isolated from biomass of an anaerobic chlorate-reducing bioreactor. Phylogenetic analysis of the 16S rDNA sequence showed 100% sequence similarity to Pseudomonas stutzeri DSM 50227 and 98.6% sequence similarity to the type strain of P. stutzeri (DSM 5190(T)). The species P. stutzeri possesses a high degree of genotypic and phenotypic heterogeneity. Therefore, eight genomic groups, termed genomovars, have been proposed based upon deltaTm values, which were used to evaluate the quality of the pairing within heteroduplexes formed by DNA-DNA hybridization. In this study, DNA-DNA hybridization between strain AW-1(T) and P. stutzeri strains DSM 50227 and DSM 5190(T) revealed respectively 80.5 and 56.5% similarity. DNA-DNA hybridization between P. stutzeri strains DSM 50227 and DSM 5190(T) revealed 48.4% similarity. DNA-DNA hybridization indicated that strain AW-1(T) is not related at the species level to the type strain of P. stutzeri. However, strain AW-1(T) and P. stutzeri DSM 50227 are related at the species level. The physiological and biochemical properties of strain AW-1(T) and the two P. stutzeri strains were compared. A common characteristic of P. stutzeri strains is the ability to denitrify. However, in growth experiments, strain AW-1(T) could use only chlorate or oxygen as an electron acceptor and not nitrate, perchlorate or bromate. Strain AW-1(T) is the first chlorate-reducing bacterium described that does not possess another oxyanion-reduction pathway. Cell extracts of strain AW-1(T) showed chlorate and bromate reductase activities but not nitrate reductase activity. P. stutzeri strains DSM 50227 and DSM 5190(T) could use nitrate or oxygen as an electron acceptor, but not chlorate. Chlorate reductase activity, in addition to nitrate reductase activity, was detected in cell extracts of both P. stutzeri strains. Chlorite dismutase activity was absent in extracts of both P. stutzeri strains but was present in extracts of strain AW-1(T). Based on the hybridization experiments and the physiological and biochemical data, it is proposed that strain AW-1(T) be classified as a novel species of Pseudomonas, Pseudomonas chloritidismutans sp. nov. The type strain is strain AW-1(T) (= DSM 13592(T) = ATCC BAA-443(T)).


Impact of long-term ticagrelor monotherapy following 1-month dual antiplatelet therapy in patients who underwent complex percutaneous coronary intervention: insights from the Global Leaders trial.
To evaluate the impact of an experimental strategy [23-month ticagrelor monotherapy following 1-month dual antiplatelet therapy (DAPT)] vs. a reference regimen (12-month aspirin monotherapy following 12-month DAPT) after complex percutaneous coronary intervention (PCI). In the present post hoc analysis of the Global Leaders trial, the primary endpoint [composite of all-cause death or new Q-wave myocardial infarction (MI)] at 2 years was assessed in patients with complex PCI, which includes at least one of the following characteristics: multivessel PCI, ≥3 stents implanted, ≥3 lesions treated, bifurcation PCI with ≥2 stents, or total stent length >60 mm. In addition, patient-oriented composite endpoint (POCE) (composite of all-cause death, any stroke, any MI, or any revascularization) and net adverse clinical events (NACE) [composite of POCE or Bleeding Academic Research Consortium (BARC) Type 3 or 5 bleeding] were explored. Among 15 450 patients included in this analysis, 4570 who underwent complex PCI had a higher risk of ischaemic and bleeding events. In patients with complex PCI, the experimental strategy significantly reduced risks of the primary endpoint [hazard ratio (HR): 0.64, 95% confidence interval (CI): 0.48-0.85] and POCE (HR: 0.80, 95% CI: 0.69-0.93), but not in those with non-complex PCI (Pinteraction = 0.015 and 0.017, respectively). The risk of BARC Type 3 or 5 bleeding was comparable (HR: 0.97, 95% CI: 0.67-1.40), resulting in a significant risk reduction in NACE (HR: 0.80, 95% CI: 0.69-0.92; Pinteraction = 0.011). Ticagrelor monotherapy following 1-month DAPT could provide a net clinical benefit for patients with complex PCI. However, in view of the overall neutral results of the trial, these findings of a post hoc analysis should be considered as hypothesis generating.


Karyotype analysis and sex determination in Australian Brush-turkeys (Alectura lathami).
Sexual differentiation across taxa may be due to genetic sex determination (GSD) and/or temperature sex determination (TSD). In many mammals, males are heterogametic (XY); whereas females are homogametic (XX). In most birds, the opposite is the case with females being heterogametic (ZW) and males the homogametic sex (ZZ). Many reptile species lack sex chromosomes, and instead, sexual differentiation is influenced by temperature with specific temperatures promoting males or females varying across species possessing this form of sexual differentiation, although TSD has recently been shown to override GSD in Australian central beaded dragons (Pogona vitticeps). There has been speculation that Australian Brush-turkeys (Alectura lathami) exhibit TSD alone and/or in combination with GSD. Thus, we sought to determine if this species possesses sex chromosomes. Blood was collected from one sexually mature female and two sexually mature males residing at Sylvan Heights Bird Park (SHBP) and shipped for karyotype analysis. Karyotype analysis revealed that contrary to speculation, Australian Brush-turkeys possess the classic avian ZW/ZZ sex chromosomes. It remains a possibility that a biased primary sex ratio of Australian Brush-turkeys might be influenced by maternal condition prior to ovulation that result in her laying predominantly Z- or W-bearing eggs and/or sex-biased mortality due to higher sensitivity of one sex in environmental conditions. A better understanding of how maternal and extrinsic factors might differentially modulate ovulation of Z- or W-bearing eggs and hatching of developing chicks possessing ZW or ZZ sex chromosomes could be essential in conservation strategies used to save endangered members of Megapodiidae.


Knowledge and perceptions of medical abortion among potential users.
Nearly two-thirds of 73 women aged 18-34 who participated in focus groups on medical abortion conducted in three cities had heard about this new abortion method, but only a few could describe it accurately. Once the method was described to them, they cited its potential advantages over vacuum aspiration as being fewer major complications, the absence of surgery, a greater "naturalness," and its use earlier in pregnancy. Women listed as disadvantages the multiple visits needed for medical abortion, the unknown aspects of the new technology, especially regarding the expulsion of the conceptus, and concern that mifepristone would make an abortion too easy and lead some women to take the decision lightly. More than one-third of discussants said they would choose mifepristone if the method were available.


A birdcage model for the Chinese meridian system: part II. The meridian system as a birdcage resonator.
We propose that the meridian system as a whole is one grand transmission line, excited periodically at a frequency of f50, the frequency of the 50-round circulation (0.578 x 10(-3) Hz). This grand transmission line is analyzed as a 28-leg, uniform, low pass (LP) electromagnetic (EM) birdcage coil, consisting of capacitors and inductors. The second lowest resonant frequency (mode 1) of the birdcage is f50. Each leg represents one channel and is analyzed as a lossless transmission line itself. The amplitude of the Qi standing wave on each channel is periodically amplified at f50. The average number of acupoints on the 28 channels involved in the 50-round circulation and the speed of Qi were used to calculate the resonant frequencies. A mechanical birdcage consisting of mass and spring may co-exist and exchange energy with the EM birdcage. Magnetic resonance imaging (MRI) may be used to map the diffusion coefficient, elastic modulus and electric conductivity of tissues in vivo, as an indirect evidence of the existence of the proposed EM and mechanical standing waves.


Molecular characterization and expression of the Erwinia carotovora hrpNEcc gene, which encodes an elicitor of the hypersensitive reaction.
The nucleotide sequence of hrpNEcc DNA, cloned from Erwinia carotovora subsp. carotovora strain Ecc71, reveals a coding region of 1,068 bp which matches the size of hrpNEcc transcripts. hrpNEcc is predicted to encode a glycine-rich protein of approximately 36 kDa. Like the elicitors of the hypersensitive reaction (HR) produced by E. chrysanthemi (HarpinEch) and E. amylovora (HarpinEa), the deduced 36-kDa protein does not possess a typical signal sequence, but it contains a putative membrane-spanning domain. In Escherichia coli strains overexpressing hrpNEcc, the 36-kDa protein has been identified as the hrpNEcc product by Western blot analysis using anti-HarpinEch antibodies. The 36-kDa protein fractionated from E. coli elicits the HR in tobacco leaves. Moreover, a HrpN- and RsmA- double mutant (RsmA = regulator of secondary metabolites) does not produce this 36-kDa protein or elicit the HR, although this strain, like the RsmA- and HrpN+ bacteria, overproduces extracellular enzymes and macerates celery petioles. These observations demonstrate that hrpNEcc encodes the elicitor of the HR, designated HarpinEcc. The levels of hrpNEcc transcripts are affected in both RsmA+ and RsmA- strains by media composition and carbon sources, although the mRNA levels are substantially higher in the RsmA- strains. The expression of hrpNEcc in Ecc71 is cell density dependent and is activated by the quorum-sensing signal, N-(3-oxohexanoyl)-L-homoserine lactone (OHL). By contrast, hrpNEcc expression in an RsmA- strain is independent of cell density, and substantial expression occurs in the absence of OHL. The effects of cultural conditions and the occurrence of putative cis-acting sequences, such as consensus sigma 54 promoters and an hrp promoter upstream of the transcriptional start site, indicate that the production of HarpinEcc in wild-type RsmA+ E. carotovora subsp. carotovora is tightly regulated. These observations, taken along with the finding that the HR is caused by RsmA- mutants but not by RsmA+ strains (Cui et al., 1996, Mol. Plant-Microbe Interact. 9:565-573), strongly support the idea that the inability of the wild-type pectolytic E. carotovora subsp. carotovora to elicit the HR is due to the lack of a significant level of HarpinEcc production.


Cerebral activations related to ballistic, stepwise interrupted and gradually modulated movements in Parkinson patients.
Patients with Parkinson's disease (PD) experience impaired initiation and inhibition of movements such as difficulty to start/stop walking. At single-joint level this is accompanied by reduced inhibition of antagonist muscle activity. While normal basal ganglia (BG) contributions to motor control include selecting appropriate muscles by inhibiting others, it is unclear how PD-related changes in BG function cause impaired movement initiation and inhibition at single-joint level. To further elucidate these changes we studied 4 right-hand movement tasks with fMRI, by dissociating activations related to abrupt movement initiation, inhibition and gradual movement modulation. Initiation and inhibition were inferred from ballistic and stepwise interrupted movement, respectively, while smooth wrist circumduction enabled the assessment of gradually modulated movement. Task-related activations were compared between PD patients (N = 12) and healthy subjects (N = 18). In healthy subjects, movement initiation was characterized by antero-ventral striatum, substantia nigra (SN) and premotor activations while inhibition was dominated by subthalamic nucleus (STN) and pallidal activations, in line with the known role of these areas in simple movement. Gradual movement mainly involved antero-dorsal putamen and pallidum. Compared to healthy subjects, patients showed reduced striatal/SN and increased pallidal activation for initiation, whereas for inhibition STN activation was reduced and striatal-thalamo-cortical activation increased. For gradual movement patients showed reduced pallidal and increased thalamo-cortical activation. We conclude that PD-related changes during movement initiation fit the (rather static) model of alterations in direct and indirect BG pathways. Reduced STN activation and regional cortical increased activation in PD during inhibition and gradual movement modulation are better explained by a dynamic model that also takes into account enhanced responsiveness to external stimuli in this disease and the effects of hyper-fluctuating cortical inputs to the striatum and STN in particular.


Nerve growth factors (NGF, BDNF) enhance axonal regeneration but are not required for survival of adult sensory neurons.
Largely on the basis of studies with nerve growth factor (NGF), it is now widely accepted that development of the peripheral nervous system of vertebrates is dependent in part on the interaction of immature sensory and autonomic neurons with specific survival factors that are derived from peripheral target fields. I have found, in marked contrast to an absolute requirement for NGF during development, that adult rat dorsal root ganglion sensory neurons are not dependent on NGF or other survival factors for long-term (3-4 weeks) maintenance in vitro. When dissociated and enriched, at least 70-80% of adult DRG neurons survived and extended long processes either in the absence of exogenously added NGF or upon the removal of any possible source of endogenous NGF or other neurotrophic activity (i.e., nonneuronal cells, in chemically defined culture medium, in the presence of an excess of anti-NGF antibodies, or when cultured as single neurons in microwells). Although not required for survival or expression of a range of complex morphologies, both NGF and brain-derived neurotrophic factor (BDNF) were found to stimulate the regeneration of axons from adult DRG neurons.


The kinetics of induction by rifampin of alpha 1-acid glycoprotein and antipyrine clearance in the dog.
Rifampin is known to be an important stimulus to drug-metabolizing enzymes and can also induce the production of alpha 1-acid glycoprotein (AGP). We have studied the time course for induction of drug metabolizing capability as assessed by the clearance of antipyrine and the plasma concentration of AGP following a chronic course of rifampin in dogs. The kinetics of the induction process were observed during a 22-day treatment period, and the wash-out period kinetics were followed for another 3 weeks. Rifampin kinetics were measured at the end of the 22-day dosing period. Both antipyrine clearance and AGP concentration were significantly increased by the rifampin treatment; antipyrine clearance doubled and AGP concentrations nearly tripled. When analyzed by a newly developed kinetic model of induction, it was determined that the time course for AGP or antipyrine clearance was not governed by a single rate constant. The second rate constant did not represent the accumulation or persistence of rifampin (t1/2 = 4.4 hr). It is hypothesized that one or more synthesis precursors to the enzymes which produce AGP or clear antipyrine are also rate limiting. This is the first example in which an induction/deinduction experiment has been interpreted from beginning to end with a three-step kinetic model. It demonstrates the applicability of this model and recommends its use in other induction experiments.


Virtual-versus-Real Implantation of Flow Diverters: Clinical Potential and Influence of Vascular Geometry.
Intracranial stents have become extremely important in the endovascular management of complex intracranial aneurysms. Sizing and landing zone predictions are still very challenging steps in the procedure. Virtual stent deployment may help therapeutic planning, device choice, and hemodynamic simulations. We aimed to assess the predictability of our recently developed virtual deployment model by comparing in vivo and virtual stents implanted in a consecutive series of patients presenting with intracranial aneurysms. Virtual stents were implanted in patient-specific geometries of intracranial aneurysms treated with the Pipeline Embolization Device. The length and cross-section of virtual and real stents measured with conebeam CT were compared. The influence of vessel geometry modifications occurring during the intervention was analyzed. The virtual deployment based on pre- and poststent implantation 3D rotational angiography overestimated (underestimated) the device length by 13% ± 11% (-9% ± 5%). These differences were highly correlated (R2 = 0.67) with the virtual-versus-real stent radius differences of -6% ± 7% (5% ± 4%) for predictions based on pre- and poststent implantation 3D rotational angiography. These mismatches were due principally to implantation concerns and vessel-shape modifications. The recently proposed geometric model was shown to predict accurately the deployment of Pipeline Embolization Devices when the stent radius was well-assessed. However, unpredictable delivery manipulations and variations of vessel geometry occurring during the intervention might impact the stent implantation.


Cost of Illness of Multiple Sclerosis - A Systematic Review.
Cost-of-illness (COI) studies of Multiple Sclerosis (MS) are vital components for describing the economic burden of MS, and are frequently used in model studies of interventions of MS. We conducted a systematic review of studies estimating the COI of MS, to compare costs between studies and examine cost drivers, emphasizing generalizability and methodological choices. A literature search on studies published in English on COI of MS was performed in PubMed for the period January 1969 to January 2014, resulting in 1,326 publications. A mapping of studies using a bottom-up approach or top-down approach, respectively, was conducted for the 48 studies assessed as relevant. In a second analysis, the cost estimates were compared between the 29 studies that used a societal perspective on costs, human capital approach for indirect costs, presenting number of patients included, time-period studied, and year of price level used. The mapping showed that bottom-up studies and prevalence approaches were most common. The cost ratios between different severity levels within studies were relatively stable, to the ratio of 1 to 2 to 3 for disability level categories. Drugs were the main cost drivers for MS-patients with low disease severity, representing 29% to 82% of all costs in this patient group, while the main cost components for groups with more advanced MS symptoms were production losses due to MS and informal care, together representing 17% to 67% of costs in those groups. The bottom-up method and prevalence approach dominated in studies of COI of MS. Our findings show that there are difficulties in comparing absolute costs across studies, nevertheless, the relative costs expressed as cost ratios, comparing different severity levels, showed higher resemblance. Costs of drugs were main cost drivers for less severe MS and informal care and production losses for the most severe MS.


"Plantas con madre": plants that teach and guide in the shamanic initiation process in the East-Central Peruvian Amazon.
We present and discuss a particular group of plants used by a diversity of healers in the initiation process and apprenticeship of traditional medicine, as practiced by Amazonian societies in East-Central Peru. Often, these plants are locally called plantas con madre (plants with a mother), and are thought to guide initiates in the process of seeking sacred knowledge, learning about plant usage, and understanding traditional medicine practices. We illustrate the diversity of plants used in the apprenticeship and practice of traditional medicine, and nurture the discussion to better understand the terminology used by Indigenous healers to describe plant uses and their practices. The study was conducted between 2003 and 2008 with the participation of 29 curanderos (healers; 23 men, 6 women), 3 apprentices and 4 herbalists. The participants belonged to four ethnic groups: 17 Mestizos, 15 Shipibo-Konibo, 1 Ashaninka, and 1 Matsiguenga; a Spanish apprentice and an Italian herbalist were also included in the study. The field data were collected using semi-structured interviews, participant observation, and the witnessing of numerous healing sessions. Oral informed consent was obtained from each participant. We identified 55 plant species belonging to 26 botanical families, which are used in initiation processes and apprenticeships of traditional medicine. This group of plants is administered under strict conditions during training and healing sessions called dietas (shamanic diets), with the supervision of one or more maestros curanderos (master healers). We observed that during the shamanic diets, maestros curanderos administered plants depending on the teachings or tools he/she was passing on, and were based on a particular sequence during the initiation process: (I) purification and cleansing species; (II) sensitivity and intuition; (III) strengthening; and (IV) protection and defence. Traditional healers continue to be a primary source of health care for the majority of the population in the Amazon region. Our research suggests that the system of dietas and the plantas con madre are fundamental components of the everyday practice of traditional medicine, maintenance of cultural continuity and Indigenous cosmovisions in the Amazonian societies in East-Central Peru. This paper contributes to filling the gap in the understanding of the process of initiation among healers in this area of the world. The study offers evidence of the need to collaborate with Indigenous healers to improve the recognition of their medical practices, role in their societies, and the value of their tools and medicines. A respectful attitude and open exchange of ideas and information may contribute to a better understanding of the language used by traditional medical practitioners, their practice, and their worldviews.


CULTIVATION OF TREPONEMA CALLIGYRUM (NEW SPECIES) FROM CONDYLOMATA OF MAN.
1. On the surface of genital or anal lesions, either syphilitic or non-syphilitic, there may be found occasionally a spirochaeta resembling Treponema pallidum, but somewhat thicker than the latter. In general characteristics, this organism occupies an intermediary position between Treponema pallidum and Spirochaeta refringens. 2. The treponema was obtained in pure culture from two cases. For the organism the name of Treponema calligyrum is proposed. 3. Treponema calligyrum is non-pathogenic for monkeys and rabbits. 4. Treponema calligyrum can be distinguished from Treponema pallidum, Treponema microdentium, Treponema macrodentium, Treponema mucosum, and Spirochaeta refringens either by the morphology, cultural and biological properties, or all these conditions in combination. 5. Under certain cultural conditions Treponema calligyrum becomes thinner and may cause some difficulty of differentiation from a strain of the thick type of Treponema pallidum, especially when the latter is grown under conditions which tend to render it thicker. There is no such difficulty when the average and thin types of Treponema pallidum are concerned. On the other hand, by providing the calligyrum with conditions which lead to the appearance of thicker specimens and by supplying Spirochaeta refringens with conditions of growth that favor the development of individuals that are thinner and more regularly curved, the refringens may be made to resemble the calligyrum in general appearance; but under identical cultural conditions they can be readily distinguished.


Lived experience of involuntary transport under mental health legislation.
Police have historically been responsible for transporting people during a mental health crisis in Australia. A major change to the New South Wales (NSW) Mental Health Act (MHA) in 2007 expanded the range of coercive transportation agencies to include NSW Ambulance (paramedics) and NSW Health (mental health nurses). Anecdotal reports, however, describe a lack of clarity around how these changes should be implemented in practice. This research aims to explore this lack of clarity through qualitative analysis of interviews with people with the lived experience of involuntary transport under the MHA. Sixteen interviews were conducted; most (n = 14) interviews in northern NSW regions: six with people who had been transported (consumers), four with carers, and six with service providers (two police, one paramedic, and three mental health nurses). For consumers and carers, the police response was often perceived as too intense, particularly if the person was not violent. Carers were often conflicted by having to call for emergency intervention. Service providers were frustrated by a lack of a coordinated interagency response, resourcing issues, delays at emergency departments, and lack of adequate training. A central theme across all groups was the importance of communication styles. As one participant (consumer) said: 'Everybody needs a lesson in kindness'. All groups agreed that high-risk situations necessitate police involvement. However, invocation of the MHA during a high-risk situation is fraught with stress and difficulties, leaving little room for empathetic communications. Effective and diverse, evidence-based, early intervention strategies - both consensual and non-consensual - are necessary to reduce the requirement for police involvement in mental health transports.


Phase correlation between four-wave mixing and optical fields in double Λ-type atomic system.
We study the spectral features and phase of four-wave mixing (FWM) light according to the relative phase-noise of the optical fields coupled to a double Λ-type atomic system of the 5S1/2-5P1/2 transition of 87Rb atoms. We observe that the spectral shape of the FWM spectrum is identical to that of the two-photon absorption (TPA) spectrum due to two-photon coherence and that it is independent of the relative phase-noise of the pump light. From these results, we clarify that the two-photon coherence plays a very important role in the FWM process. Furthermore, we measure the relative linewidth of the FWM signal to the probe and pump lasers by means of a beat interferometer. We confirmed that the phase of the FWM signal is strongly correlated with that of the pump laser under the condition of phase-locked probe and coupling lasers for two-photon coherence.


The ACGME competencies in the operating room.
In 1999, the ACGME introduced the 6 competencies that have become the basis for resident education. Since the operating room (OR) has traditionally been the major focus for resident teaching in surgery, we performed an observational study to determine whether it is an appropriate setting for the teaching and/or assessment of the competencies. A 3-person team observed 11 operations and recorded all teaching events. Observers then determined whether each event involved the teaching of a competency by faculty or demonstration of a competency by residents. Frequency counts, mean times, and ranges were calculated for each competency taught and demonstrated. The Patient Care competency was both the most commonly taught and demonstrated. Faculty spent an average of 33% of operative time instructing in patient care, and residents demonstrated it 65% of the time. The Interpersonal/Communication Skills (4%) and Practice-Based Learning/Improvement (4%) competencies were also occasionally demonstrated by residents. The remaining competencies were addressed less frequently. OR teaching was primarily devoted to the Patient Care competency. The OR was also an appropriate setting for evaluating resident performance in this area. New approaches to OR teaching or educational efforts in other settings such as the clinic are necessary for teaching and assessing the remaining competencies.


Interleukin role in the regulation of endothelial cell pathological activation.
Interleukins (ILs) are the group of cytokines firstly identified as expressed by leukocytes and playing different immunomodulatory functions. With increasing evidence of a constant crosstalk between leukocytes and endothelial cells in the regulation of immune cell differentiation and activation, a role of ILs also in endothelial cell stimulation and vascular inflammation has been shown. ILs act on endothelial cells both in an autocrine and a paracrine manner. In fact, a cross regulation is present among ILs expressed by different cell types, leading to amplification or blocking of the initial inflammatory signal with the secretion of additional ILs or involvement of other adjacent cells and tissues. Based on selective structural features, ILs can be divided into four major groups, a fifth group comprises ILs that do not fit into any of the other four. Most of the ILs playing a role in endothelial cell activation belong to the IL1-like cytokine group, but the number of ILs involved in vascular inflammation is constantly growing, and a special contribution of IL6, IL8, and IL17 has been underlined. This review aims at presenting current knowledge and at underling missing information about the role of IL in activating endothelial cells in selected pathological settings such as tumours, psoriasis, systemic sclerosis, and viral infection.


A new synthesis in epigenetics: towards a unified function of DNA methylation from invertebrates to vertebrates.
DNA methylation is generally limited to CpG doublets located at the gene promoter with an involvement in gene silencing. Surprisingly, two recent papers showed an extensive methylation affecting coding portions of transcriptionally active genes in human and plants prompting a rethink of DNA methylation in eukaryotes. Actually, gene body methylation is not surprising since it has been repeatedly reported in invertebrates, where it interferes with transcriptional elongation preventing aberrant transcription initiations. As a whole, the published data suggest that the most ancestral function of DNA methylation is the control of genes that are susceptible to transcriptional interference and not to gene silencing. The recruitment of DNA methylation for silencing represents a successive tinkered use. In view of this additional function, the invertebrate-vertebrate transition has been accompanied by new constraints on DNA methylation that resulted in the strong conservation of the DNA methylation machinery in vertebrates and in the non-viability of mutants lacking DNA methylation.


Evaluation of blood culture procedures in a pediatric hospital.
To determine optimal clinical laboratory techniques for detecting pediatric bacteremia, we studied 7,768 consecutive blood cultures in a 1-year period. Blood was inoculated into one vented 50-ml bottle of brucella broth with 0.05% sodium polyanetholsulfonate and one unvented 50-ml bottle of Columbia broth with 0.05% sodium polyanetholsulfonate and 0.05% cysteine. Bottles were visually examined for growth on days 1 through 7 and blindly subcultured aerobically and anaerobically on days 1, 2, and 7. There were 724 (9.3%) positive cultures, and 484 (6.2%) were clinically significant. The most frequent isolates from bacteremic patients were Haemophilus influenzae (24%) and Streptococcus pneumoniae (17%). Growth was noted in only one bottle in 25% of clinically significant isolates. Bottles inoculated with greater than or equal to 1 ml of blood became positive earlier than bottles inoculated with less than 1 ml. After 1 day of incubation, 48% of the clinically significant cultures showed growth on visual examination, whereas 85% showed growth on subculture. Only 19% of Haemophilus isolates were detected visually on day 1, whereas 88% were recovered on subculture. By day 7, 3.5% of all isolates (including 18% of pneumococcal isolates and 1% of Haemophilus isolates) could no longer be recovered on subculture. We conclude that a two-bottle blood culture system and blind subculture within 24 h will optimize detection of pediatric bacteremia.


Virus susceptibility of Chinese hamster ovary (CHO) cells and detection of viral contaminations by adventitious agent testing.
Biopharmaceuticals are of increasing importance in the treatment of a variety of diseases. A remaining concern associated with their production is the potential introduction of adventitious agents into their manufacturing process, which may compromise the pathogen safety of a product and potentially cause stock-out situations for important medical supplies. To ensure the safety of biological therapeutics, regulatory guidance requires adventitious agent testing (AAT) of the bulk harvest. AAT is a deliberately promiscuous assay procedure which has been developed to indicate, ideally, the presence of any viral contaminant. One of the most important cell lines used in the production of biopharmaceuticals is Chinese hamster ovary (CHO) cells and while viral infections of CHO cells have occurred, a systematic screen of their virus susceptibility has never been published. We investigated the susceptibility of CHO cells to infection by 14 different viruses, including members of 12 families and representatives or the very species that were implicated in previously reported production cell infections. Based on our results, four different infection outcomes were distinguished, based on the possible combinations of the two factors (i) the induction, or not, of a cytopathic effect and (ii) the ability, or not, to replicate in CHO cells. Our results demonstrate that the current AAT is effective for the detection of viruses which are able to replicate in CHO cells. Due to the restricted virus susceptibility of CHO cells and the routine AAT of bulk harvests, our results provide re-assurance for the very high safety margins of CHO cell-derived biopharmaceuticals.


Design and Analysis of the 4-Anilinoquin(az)oline Kinase Inhibition Profiles of GAK/SLK/STK10 Using Quantitative Structure-Activity Relationships.
The 4-anilinoquinoline and 4-anilinoquinazoline ring systems have been the focus of significant efforts in prior kinase drug discovery programs, which have led to approved medicines. Broad kinome profiles of these compounds have now been assessed with the advent of advanced screening technologies. These ring systems, while originally designed for specific targets including epidermal growth factor receptor (EGFR), but actually display a number of potent collateral kinase targets, some of which have been associated with negative clinical outcomes. We have designed and synthesized a series of 4-anilinoquin(az)olines in order to better understand the structure-activity relationships of three main collateral kinase targets of quin(az)oline-based kinase inhibitors: cyclin G associated kinase (GAK), STE20-like serine/threonine-protein kinase (SLK) and serine/threonine-protein kinase 10 (STK10). This was achieved through a series of quantitative structure-activity relationship (QSAR) analysis, water mapping of the kinase ATP binding sites and extensive small-molecule X-ray structural analysis.


Communication of BRCA1 and BRCA2 genetic test results to health care providers following genetic testing at a tertiary care center.
Individuals at high risk for hereditary cancers often receive genetic counseling and testing at tertiary care centers; however, they may receive care for long-term management of their cancer risk in community settings. Communication of genetic test results to health care providers outside of tertiary care settings can facilitate the long-term management of high risk individuals. This study assessed women's communication of BRCA1/BRCA2 genetic test results to health care providers outside of tertiary care settings (termed "outside" health care providers, or OHCPs) and women's perceptions regarding communication of results. Women (n = 312) who underwent BRCA1/BRCA2 genetic counseling and testing completed a questionnaire assessing whether or not they shared test results with OHCPs and perceptions regarding the communication of test results to OHCPs. Most (72%) shared genetic test results with OHCPs. Women with no personal history of cancer were more likely to have shared results compared to women with a personal history of cancer. Mutation status did not significantly predict sharing of genetic information. Most reported positive perceptions regarding the disclosure of genetic test results to OHCPs. The majority did not report any concerns about potential insurance discrimination (88%) and indicated that OHCPs were able to appropriately address their questions (81%). Although most women shared their genetic test results with OHCPs, those with a personal history of cancer may need further encouragement to share this information. Tertiary care centers should facilitate outreach and education with OHCPs in order to assure appropriate long-term cancer risk management for high risk populations.


BayCANN: Streamlining Bayesian Calibration With Artificial Neural Network Metamodeling.
Purpose: Bayesian calibration is generally superior to standard direct-search algorithms in that it estimates the full joint posterior distribution of the calibrated parameters. However, there are many barriers to using Bayesian calibration in health decision sciences stemming from the need to program complex models in probabilistic programming languages and the associated computational burden of applying Bayesian calibration. In this paper, we propose to use artificial neural networks (ANN) as one practical solution to these challenges. Methods: Bayesian Calibration using Artificial Neural Networks (BayCANN) involves (1) training an ANN metamodel on a sample of model inputs and outputs, and (2) then calibrating the trained ANN metamodel instead of the full model in a probabilistic programming language to obtain the posterior joint distribution of the calibrated parameters. We illustrate BayCANN using a colorectal cancer natural history model. We conduct a confirmatory simulation analysis by first obtaining parameter estimates from the literature and then using them to generate adenoma prevalence and cancer incidence targets. We compare the performance of BayCANN in recovering these "true" parameter values against performing a Bayesian calibration directly on the simulation model using an incremental mixture importance sampling (IMIS) algorithm. Results: We were able to apply BayCANN using only a dataset of the model inputs and outputs and minor modification of BayCANN's code. In this example, BayCANN was slightly more accurate in recovering the true posterior parameter estimates compared to IMIS. Obtaining the dataset of samples, and running BayCANN took 15 min compared to the IMIS which took 80 min. In applications involving computationally more expensive simulations (e.g., microsimulations), BayCANN may offer higher relative speed gains. Conclusions: BayCANN only uses a dataset of model inputs and outputs to obtain the calibrated joint parameter distributions. Thus, it can be adapted to models of various levels of complexity with minor or no change to its structure. In addition, BayCANN's efficiency can be especially useful in computationally expensive models. To facilitate BayCANN's wider adoption, we provide BayCANN's open-source implementation in R and Stan.


A simple versatile solution for collecting multidimensional clinical data based on the CakePHP web application framework.
Clinical trials aiming for regulatory approval of a therapeutic agent must be conducted according to Good Clinical Practice (GCP). Clinical Data Management Systems (CDMS) are specialized software solutions geared toward GCP-trials. They are however less suited for data management in small non-GCP research projects. For use in researcher-initiated non-GCP studies, we developed a client-server database application based on the public domain CakePHP framework. The underlying MySQL database uses a simple data model based on only five data tables. The graphical user interface can be run in any web browser inside the hospital network. Data are validated upon entry. Data contained in external database systems can be imported interactively. Data are automatically anonymized on import, and the key lists identifying the subjects being logged to a restricted part of the database. Data analysis is performed by separate statistics and analysis software connecting to the database via a generic Open Database Connectivity (ODBC) interface. Since its first pilot implementation in 2011, the solution has been applied to seven different clinical research projects covering different clinical problems in different organ systems such as cancer of the thyroid and the prostate glands. This paper shows how the adoption of a generic web application framework is a feasible, flexible, low-cost, and user-friendly way of managing multidimensional research data in researcher-initiated non-GCP clinical projects.


Health Informatics for Neonatal Intensive Care Units: An Analytical Modeling Perspective.
The effective use of data within intensive care units (ICUs) has great potential to create new cloud-based health analytics solutions for disease prevention or earlier condition onset detection. The Artemis project aims to achieve the above goals in the area of neonatal ICUs (NICU). In this paper, we proposed an analytical model for the Artemis cloud project which will be deployed at McMaster Children's Hospital in Hamilton. We collect not only physiological data but also the infusion pumps data that are attached to NICU beds. Using the proposed analytical model, we predict the amount of storage, memory, and computation power required for the system. Capacity planning and tradeoff analysis would be more accurate and systematic by applying the proposed analytical model in this paper. Numerical results are obtained using real inputs acquired from McMaster Children's Hospital and a pilot deployment of the system at The Hospital for Sick Children (SickKids) in Toronto.


Polarity mediates asymmetric trafficking of the Gbeta heterotrimeric G-protein subunit GPB-1 in C. elegans embryos.
Asymmetric cell division is an evolutionarily conserved process that gives rise to daughter cells with different fates. In one-cell stage C. elegans embryos, this process is accompanied by asymmetric spindle positioning, which is regulated by anterior-posterior (A-P) polarity cues and driven by force generators located at the cell membrane. These force generators comprise two Gα proteins, the coiled-coil protein LIN-5 and the GoLoco protein GPR-1/2. The distribution of GPR-1/2 at the cell membrane is asymmetric during mitosis, with more protein present on the posterior side, an asymmetry that is thought to be crucial for asymmetric spindle positioning. The mechanisms by which the distribution of components such as GPR-1/2 is regulated in time and space are incompletely understood. Here, we report that the distribution of the Gβ subunit GPB-1, a negative regulator of force generators, varies across the cell cycle, with levels at the cell membrane being lowest during mitosis. Furthermore, we uncover that GPB-1 trafficks through the endosomal network in a dynamin- and RAB-5-dependent manner, which is most apparent during mitosis. We find that GPB-1 trafficking is more pronounced on the anterior side and that this asymmetry is regulated by A-P polarity cues. In addition, we demonstrate that GPB-1 depletion results in the loss of GPR-1/2 asymmetry during mitosis. Overall, our results lead us to propose that modulation of Gβ trafficking plays a crucial role during the asymmetric division of one-cell stage C. elegans embryos.


The CMAP scan as a tool to monitor disease progression in ALS and PMA.
Amyotrophic lateral sclerosis (ALS) and progressive muscular atrophy (PMA) are characterized by a loss of motor units (MUs), reinnervation and, eventually, muscle fibre loss. These three aspects are all reflected in the compound muscle action potential scan (CMAP scan, a high-detail stimulus response curve), which visualizes large MU potentials as 'steps'. We explored changes in the CMAP scan over time, combined the information on steps and CMAP amplitude into a CMAP scan-based progression score (CSPS), and correlated this score with motor unit number estimates (MUNE). Ten patients (three PMA, seven ALS; age 37-77 years) were included. CMAP scan and MUNE measurements were performed five times during a three-month period. Nine patients had additional measurements. The follow-up period was 3-24 months. Results demonstrated that abnormalities in steps preceded a decline in maximum CMAP amplitude during follow-up. Usually, both steps and maximum CMAP amplitude changed between recordings. The correlation between the CSPS and MUNE was -0.80 (p < 0.01). In conclusion, the CMAP scan can be used to visualize and quantify disease progression in a muscle affected by MND. The CSPS is a measure of MU loss that is quick and easy to obtain and that, in contrast to MUNE, has no sample bias.


Fluoride Rinses, Gels and Foams: An Update of Controlled Clinical Trials.
The aim of this conference paper was to systematically review the quality of evidence and summarize the findings of clinical trials published after 2002 using fluoride mouth rinses, fluoride gels or foams for the prevention of dental caries. Relevant papers were selected after an electronic search for literature published in English between 2003 and 2014. The included papers were assessed for their risk of bias and the results were narratively synthesized due to study heterogeneity. The quality of evidence was expressed according to GRADE. A total of 19 papers were included (6 on fluoride mouth rinse, 10 on fluoride gel and 3 on fluoride foam); 6 had a low risk of bias while 2 had a moderate risk. All fluoride measures appeared to be beneficial in preventing crown caries and reversing root caries, but the quality of evidence was graded as low for fluoride mouth rinse, moderate for fluoride gel and very low for acidulated fluoride foam. No conclusions could be drawn on the cost-effectiveness. This review, covering the recent decade, has further substantiated the evidence for a caries-preventive effect of fluoride mouth rinse, fluoride gel and foam, previously established in systematic reviews. The lack of clinical trials free from bias is, however, still a concern, especially for fluoride mouth rinses and fluoride foam. There is also a scientific knowledge gap on the benefit and optimal use of these fluoride supplements in combination with daily tooth brushing with fluoride toothpaste.


Incentives and barriers to neurological clinical research participation.
BACKGROUND: Understanding barriers and incentives to neurological research participation may improve clinical trial enrollment. RESULTS: Telephone-based focus groups were conducted with four neurological disorder patient groups (n = 22) and one caregiver group (n = 6). A total of 14 neurologists and neurosurgeons participated in structured interviews. Topics discussed included identifying ways in which health information is gathered and attitudes toward participation in medical research. Interestingly, 86% of physicians interviewed have referred patients to clinical trials and 82% of patients expressed interest in clinical trials participation. Patients cited their primary physician as the best source of health-related information, including information about clinical trials. Barriers to patients and physicians regarding clinical trials participation included compensation. Patients expressed concern about increased visit frequency and required treatment changes. CONCLUSION: Patients are willing to participate in clinical trials and physicians are willing to refer patients for participation with appropriate compensation for their time and sensitivity to change in care.


Hydrogen peroxide inhibition of bicupin oxalate oxidase.
Oxalate oxidase is a manganese containing enzyme that catalyzes the oxidation of oxalate to carbon dioxide in a reaction that is coupled with the reduction of oxygen to hydrogen peroxide. Oxalate oxidase from Ceriporiopsis subvermispora (CsOxOx) is the first fungal and bicupin enzyme identified that catalyzes this reaction. Potential applications of oxalate oxidase for use in pancreatic cancer treatment, to prevent scaling in paper pulping, and in biofuel cells have highlighted the need to understand the extent of the hydrogen peroxide inhibition of the CsOxOx catalyzed oxidation of oxalate. We apply a membrane inlet mass spectrometry (MIMS) assay to directly measure initial rates of carbon dioxide formation and oxygen consumption in the presence and absence of hydrogen peroxide. This work demonstrates that hydrogen peroxide is both a reversible noncompetitive inhibitor of the CsOxOx catalyzed oxidation of oxalate and an irreversible inactivator. The build-up of the turnover-generated hydrogen peroxide product leads to the inactivation of the enzyme. The introduction of catalase to reaction mixtures protects the enzyme from inactivation allowing reactions to proceed to completion. Circular dichroism spectra indicate that no changes in global protein structure take place in the presence of hydrogen peroxide. Additionally, we show that the CsOxOx catalyzed reaction with the three carbon substrate mesoxalate consumes oxygen which is in contrast to previous proposals that it catalyzed a non-oxidative decarboxylation with this substrate.


Community based weighing of newborns and use of mobile phones by village elders in rural settings in Kenya: a decentralised approach to health care provision.
Identifying every pregnancy, regardless of home or health facility delivery, is crucial to accurately estimating maternal and neonatal mortality. Furthermore, obtaining birth weights and other anthropometric measurements in rural settings in resource limited countries is a difficult challenge. Unfortunately for the majority of infants born outside of a health care facility, pregnancies are often not recorded and birth weights are not accurately known. Data from the initial 6 months of the Maternal and Neonatal Health (MNH) Registry Study of the Global Network for Women and Children's Health study area in Kenya revealed that up to 70% of newborns did not have exact weights measured and recorded by the end of the first week of life; nearly all of these infants were born outside health facilities. To more completely obtain accurate birth weights for all infants, regardless of delivery site, village elders were engaged to assist in case finding for pregnancies and births. All elders were provided with weighing scales and mobile phones as tools to assist in subject enrollment and data recording. Subjects were instructed to bring the newborn infant to the home of the elder as soon as possible after birth for weight measurement.The proportion of pregnancies identified before delivery and the proportion of births with weights measured were compared before and after provision of weighing scales and mobile phones to village elders. Primary outcomes were the percent of infants with a measured birth weight (recorded within 7 days of birth) and the percent of women enrolled before delivery. The recorded birth weight increased from 43 ± 5.7% to 97 ± 1.1. The birth weight distributions between infants born and weighed in a health facility and those born at home and weighed by village elders were similar. In addition, a significant increase in the percent of subjects enrolled before delivery was found. Pregnancy case finding and acquisition of birth weight information can be successfully shifted to the community level.


Mechanisms underlying capsaicin effects in canine coronary artery: implications for coronary spasm.
The TRPV1, transient receptor potential vanilloid type 1, agonist capsaicin is considered to be beneficial for cardiovascular health because it dilates coronary arteries through an endothelial-dependent mechanism and may slow atheroma progression. However, recent reports indicate that high doses of capsaicin may constrict coronary arterioles and even provoke myocardial infarction. Thus far, the mechanisms by which TRPV1 activation modulates coronary vascular tone remain poorly understood. This investigation examined whether there is a synergistic interplay between locally acting vasoconstrictive pro-inflammatory hormones (autacoids) and capsaicin effects in the coronary circulation. Experiments were performed in canine conduit coronary artery rings and isolated smooth muscle cells (CASMCs). Isometric tension measurements revealed that 1-10 μM capsaicin alone did not affect resting tension of coronary artery rings. In contrast, in endothelium-intact rings pre-contracted with a Gq/11-coupled FP/TP (prostaglandin F/thromboxane) receptor agonist, prostaglandin F2α (PGF2α; 10 μM), capsaicin first induced transient dilation that was followed by sustained contraction. In endothelium-denuded rings pre-contracted with PGF2α or thromboxane analogue U46619 (1 μM, a TP receptor agonist), capsaicin induced only sustained contraction. Blockers of the TP receptor or TRPV1 significantly inhibited capsaicin effects, but these were still observed in the presence of 50 μM nifedipine and 70 mM KCl. Capsaicin also potentiated 20 mM KCl-induced contractions. Fluorescence imaging experiments in CASMCs revealed that the Gq/11-phospholipase C (PLC)-protein kinase C (PKC) and Ca(2+)-PLC-PKC pathways are likely involved in sensitizing CASMC TRPV1 channels. Capsaicin alone does not cause contractions in conduit canine coronary artery; however, pre-treatment with pro-inflammatory prostaglandin-thromboxane agonists may unmask capsaicin's vasoconstrictive potential.


Soccer injuries of youth.
During a soccer tournament with participation of 6,600 boys and girls (9-19 years) all injuries were evaluated, 5.2% of the players were injured; out of these 51% had "slight injuries", 42% "moderate injuries" and 7% "severe injuries". For the individual player the incidence of injury was 19.1/1,000 playing hours including all degrees of injuries; if "slight injuries" were excluded the incidence was 9.4/1,000 playing hours. The incidence rose with increasing age, girls were injured more often than boys. There were 81% of all injuries localised to the lower extremity, especially the ankle and foot. Contusion was the most frequent diagnosis, amounting to a third of all injuries. Blisters and exoriations amounted to nearly a fifth, 4% of the injuries were fractures, especially in the upper extremity; overuse injuries were seen only in 5.2% of the cases. Based on examination of the injury pattern in these children, injuries of youth seem to be relatively rare and mostly of a non-severe character.


Awareness of necessity to call 9-1-1 for stroke symptoms, upstate New York.
Stroke is the third leading cause of death and a leading cause of disability in New York State. A New York study determined that only 19.9% of patients arrived at a designated stroke center within 3 hours of symptom onset. Yet, receiving treatment within 90 minutes of stroke symptom onset is optimal for improved outcomes. Delay in recognition of stroke symptoms and their severity contributes to treatment delay. A random-digit-dialed, list-assisted telephone survey about stroke knowledge was administered to 1789 adults aged 30 years or older in upstate New York in 2006. Bivariate and regression analysis were used to examine factors associated with intent to call 9-1-1 for symptoms of stroke. The largest proportion of respondents (72.4%; 95% confidence interval [CI], 69.9%-74.8%) reported they would call 9-1-1 if they noticed they or someone else had difficulty speaking, and the fewest (33.3%; 95% CI, 30.7%-36.0%) respondents reported they would call 9-1-1 for trouble seeing or double vision. Multivariate analysis found that those who had a history of delay in getting medical care in the past 6 months had decreased odds of intending to call 9-1-1 for stroke symptoms (difficulty speaking: adjusted odds ratio [AOR], 0.76; 95% CI, 0.58-1.00; trouble seeing: AOR, 0.69; 95% CI, 0.53-0.91; facial droop: AOR, 0.85; 95% CI, 0.65-1.11; arm weakness: AOR, 0.80; 95% CI, 0.63-1.03). Age, education, and history of a stroke or heart event were not consistently associated with intent to call 9-1-1. Survey respondents do not interpret some stroke symptoms as urgent enough to activate the emergency medical system. History of delaying care is a behavioral pattern that influenced intent to call 9-1-1.


Development and validation of the WASP classification system for optical diagnosis of adenomas, hyperplastic polyps and sessile serrated adenomas/polyps.
Accurate endoscopic differentiation would enable to resect and discard small and diminutive colonic lesions, thereby increasing cost-efficiency. Current classification systems based on narrow band imaging (NBI), however, do not include neoplastic sessile serrated adenomas/polyps (SSA/Ps). We aimed to develop and validate a new classification system for endoscopic differentiation of adenomas, hyperplastic polyps and SSA/Ps <10 mm. We developed the Workgroup serrAted polypS and Polyposis (WASP) classification, combining the NBI International Colorectal Endoscopic classification and criteria for differentiation of SSA/Ps in a stepwise approach. Ten consultant gastroenterologists predicted polyp histology, including levels of confidence, based on the endoscopic aspect of 45 polyps, before and after participation in training in the WASP classification. After 6 months, the same endoscopists predicted polyp histology of a new set of 50 polyps, with a ratio of lesions comparable to daily practice. The accuracy of optical diagnosis was 0.63 (95% CI 0.54 to 0.71) at baseline, which improved to 0.79 (95% CI 0.72 to 0.86, p<0.001) after training. For polyps diagnosed with high confidence the accuracy was 0.73 (95% CI 0.64 to 0.82), which improved to 0.87 (95% CI 0.80 to 0.95, p<0.01). The accuracy of optical diagnosis after 6 months was 0.76 (95% CI 0.72 to 0.80), increasing to 0.84 (95% CI 0.81 to 0.88) considering high confidence diagnosis. The combined negative predictive value with high confidence of diminutive neoplastic lesions (adenomas and SSA/Ps together) was 0.91 (95% CI 0.83 to 0.96). We developed and validated the first integrative classification method for endoscopic differentiation of small and diminutive adenomas, hyperplastic polyps and SSA/Ps. In a still image evaluation setting, introduction of the WASP classification significantly improved the accuracy of optical diagnosis overall as well as SSA/P in particular, which proved to be sustainable after 6 months.


Tbx5 is essential for heart development.
Mutations in the Tbx5 transcription factor cause heart septal defects found in human Holt-Oram Syndrome. The complete extent to which Tbx5 functions in heart development, however, has not been established. Here we show that, in Xenopus embryos, Tbx5 is expressed in the early heart field, posterior to the cardiac homeobox transcription factor, Nkx2.5. During morphogenesis, Tbx5 is expressed throughout the heart tube except the anterior portion, the bulbus cordis. When Tbx5 activity is antagonized with a hormone-inducible, dominant negative version of the protein, the heart fails to develop. These results suggest that, in addition to its function in heart septation, Tbx5 has a more global role in cardiac specification and heart development in vertebrate embryos.


Role of tryptophan residues in gramicidin channel organization and function.
The linear peptide gramicidin forms prototypical ion channels specific for monovalent cations and has been used extensively to study the organization, dynamics, and function of membrane-spanning channels. The tryptophan residues in gramicidin channels are crucial for maintaining the structure and function of the channel. We explored the structural basis for the reduction in channel conductance in the case of single-tryptophan analogs of gramicidin with three Trp --> hydrophobic substitutions using a combination of fluorescence approaches, which include red edge excitation shift and membrane penetration depth analysis, size-exclusion chromatography, and circular dichroism spectroscopy. We show here that the gramicidin analogs containing single-tryptophan residues adopt a mixture of nonchannel and channel conformations, as evident from analysis of membrane penetration depth, size-exclusion chromatography, and backbone circular dichroism data. These results are potentially useful in analyzing the effect of tryptophan substitution on the functioning of other ion channels and membrane proteins.


Involvement of the intrinsic/default system in movement-related self recognition.
The question of how people recognize themselves and separate themselves from the environment and others has long intrigued philosophers and scientists. Recent findings have linked regions of the 'default brain' or 'intrinsic system' to self-related processing. We used a paradigm in which subjects had to rely on subtle sensory-motor synchronization differences to determine whether a viewed movement belonged to them or to another person, while stimuli and task demands associated with the "responded self" and "responded other" conditions were precisely matched. Self recognition was associated with enhanced brain activity in several ROIs of the intrinsic system, whereas no differences emerged within the extrinsic system. This self-related effect was found even in cases where the sensory-motor aspects were precisely matched. Control conditions ruled out task difficulty as the source of the differential self-related effects. The findings shed light on the neural systems underlying bodily self recognition.


A case report of conservative management of extensive proliferative verrucous leukoplakia using a carbon dioxide laser.
The purpose of this report was to describe a clinical case of conservative management of extensive proliferative verrucous leukoplakia (PVL) using a carbon dioxide laser, as an alternative to complete glossectomy, which led to a sustained successful outcome over a 12-year follow-up period. Patients with oral PVL, a high risk premalignant condition, are often treated by wide surgical excision. This approach can lead to significant morbidity and compromised quality of life. A 50-year-old male with PVL and superficially invasive carcinoma, who had previously refused a complete glossectomy, presented for treatment. A baseline MRI and careful clinical evaluation by symptom review, inspection, palpation, toluidine blue staining, and biopsy were performed at baseline and at each follow-up to rule out invasive carcinoma. The PVL was treated on an outpatient basis by serial carbon dioxide laser excisions and/or vaporizations. This approach resulted in good clinical response and decreased morbidity over a 12-year follow-up period, as compared with the option of wide surgical excision. This case illustrates the benefit of a more conservative treatment approach than wide surgical excision for PVL with superficially invasive carcinoma. This approach requires greater skills, training, and experience in the evaluation of oral premalignant or malignant changes. However, decreased long-term morbidity and improved quality of life make it a valuable treatment option for PVL.