T1DBase: update 2011, organization and presentation of large-scale data sets for type 1 diabetes research.
T1DBase (http://www.t1dbase.org) is web platform, which supports the type 1 diabetes (T1D) community. It integrates genetic, genomic and expression data relevant to T1D research across mouse, rat and human and presents this to the user as a set of web pages and tools. This update describes the incorporation of new data sets, tools and curation efforts as well as a new website design to simplify site use. New data sets include curated summary data from four genome-wide association studies relevant to T1D, HaemAtlas-a data set and tool to query gene expression levels in haematopoietic cells and a manually curated table of human T1D susceptibility loci, incorporating genetic overlap with other related diseases. These developments will continue to support T1D research and allow easy access to large and complex T1D relevant data sets.


Animal and culture models of glaucoma for studying neuroprotection.
Neuroprotection aims to treat nervous system disease by maintaining the health and function of neurons. The final proof of the neuroprotective strategy relies on randomized, controlled clinical trials, but the choice of which agents to study for these trials depends on studies in the laboratory using culture and animal models. Most culture models for studying ocular neuroprotection use retinal cells, and a range of mechanisms can be studied in culture, e.g. axotomy and serum or growth factor deprivation. A variety of animal models are available for studying neuroprotection as possible therapy for glaucomatous optic neuropathy. Those most closely related to glaucoma are probably associated with moderate elevation of the intraocular pressure to levels similar to those seen in patients with untreated glaucomatous optic neuropathy. Care should be taken when applying the results of these models to humans, and there is no single criterion for deciding which culture or animal model is most relevant to the clinical situation. The most important feature is whether the model's results correlate with clinical results, and this information will only become available over time, as randomized clinical trials are completed.


Get off my back! Rapid receptor internalization through circular dorsal ruffles.
Internalization and subsequent trafficking of receptor tyrosine kinases (RTKs) play an important role in the modulation of growth factor-stimulated signaling events that affect different cellular processes, from cell growth and mitosis to motility and invasion. The intracellular transport of these receptors has traditionally been viewed as being initiated via clathrin-coated pits. However, nonclathrin pathways have been implicated as well, although these remain poorly understood. Most recently, the formation of dynamic, transient endocytic membrane structures termed circular dorsal ruffles or "dorsal waves" have been reported to selectively sequester and internalize a large percentage of a specific RTK from the surface of growth factor-stimulated cells. This process is dependent on dynamin and cortactin, two endocytic proteins that are also associated with the actin cytoskeleton, whereas it is independent of traditional coat proteins, such as clathrin and caveolin. Additionally, dorsal wave formation requires the participation and remodeling of a dynamic actin cytoskeleton. Most importantly, the formation of these structures may be less frequent in tumor cells and thereby have significant effects on receptor signaling and cell growth.


The effect of urologist experience on choosing active surveillance for prostate cancer.
To evaluate the impact of the urologist's experience in selecting active surveillance (AS) versus immediate treatment (IT) for low-risk prostate cancer. Men with low-risk prostate cancer were enrolled from March 2011 to August 2013 at 13 medical centers in Kaiser Permanente Southern California. The AS cohort was defined as men who had cT1-T2a stage prostate cancer, prostate-specific antigen <10 ng/ml, a biopsy revealing Gleason grade ≤6, fewer than three biopsy cores positive, ≤50 % cancer in any core, and not undergone immediate therapy (surgery, radiation, other) within 6 months following diagnosis. The urologist's experience (age, number of years in practice, number of robotic surgeries performed, and fellowship experience in oncology and/or robotics) was then compared between AS and IT cohorts. A total of 4754 men were diagnosed with prostate cancer, and 713 men satisfied with inclusion criteria; 433 (60.7 %) and 280 (39.3 %) chose AS and IT, respectively. A total of 87 urologists were included. Univariate and multivariate adjusted analyses revealed no differences in urologist's age or years in practice. Patients who saw urologists who had performed ≥50 robotic surgeries were less likely to choose AS (OR 0.40, 95 % CI 0.25-0.66). Patients who saw urologists with a fellowship in oncology and/or robotics were more than twice as likely to choose AS (OR 2.27, 95 % CI 1.38-3.75). These data suggest that the decision to pursue AS may be influenced by the urologist's experience.


New approaches in antimalarial drug discovery and development: a review.
Malaria remains a major world health problem following the emergence and spread of Plasmodium falciparum that is resistant to the majority of antimalarial drugs. This problem has since been aggravated by a decreased sensitivity of Plasmodium vivax to chloroquine. This review discusses strategies for evaluating the antimalarial activity of new compounds in vitro and in animal models ranging from conventional tests to the latest high-throughput screening technologies. Antimalarial discovery approaches include the following: the discovery of antimalarials from natural sources, chemical modifications of existing antimalarials, the development of hybrid compounds, testing of commercially available drugs that have been approved for human use for other diseases and molecular modelling using virtual screening technology and docking. Using these approaches, thousands of new drugs with known molecular specificity and active against P. falciparum have been selected. The inhibition of haemozoin formation in vitro, an indirect test that does not require P. falciparum cultures, has been described and this test is believed to improve antimalarial drug discovery. Clinical trials conducted with new funds from international agencies and the participation of several industries committed to the eradication of malaria should accelerate the discovery of drugs that are as effective as artemisinin derivatives, thus providing new hope for the control of malaria.


Optical coherence tomography in parkinsonian syndromes.
Parkinson's disease (PD) and the atypical parkinsonian syndromes multiple system atrophy (MSA), progressive supranuclear palsy (PSP) and corticobasal syndrome (CBS) are movement disorders associated with degeneration of the central nervous system. Degeneration of the retina has not been systematically compared in these diseases. This cross-sectional study used spectral-domain optical coherence tomography with manual segmentation to measure the peripapillar nerve fiber layer, the macular thickness, and the thickness of all retinal layers in foveal scans of 40 patients with PD, 19 with MSA, 10 with CBS, 15 with PSP, and 35 age- and sex-matched controls. The mean paramacular thickness and volume were reduced in PSP while the mean RNFL did not differ significantly between groups. In PSP patients, the complex of retinal ganglion cell- and inner plexiform layer and the outer nuclear layer was reduced. In PD, the inner nuclear layer was thicker than in controls, MSA and PSP. Using the ratio between the outer nuclear layer and the outer plexiform layer with a cut-off at 3.1 and the additional constraint that the inner nuclear layer be under 46 µm, we were able to differentiate PSP from PD in our patient sample with a sensitivity of 96% and a specificity of 70%. Different parkinsonian syndromes are associated with distinct changes in retinal morphology. These findings may serve to facilitate the differential diagnosis of parkinsonian syndromes and give insight into the degenerative processes of patients with atypical parkinsonian syndromes.


Legionnaires' Disease in Hotels and Passenger Ships: A Systematic Review of Evidence, Sources, and Contributing Factors.
Travel-associated Legionnaires' disease (LD) is a serious problem, and hundreds of cases are reported every year among travelers who stayed at hotels, despite the efforts of international and governmental authorities and hotel operators to prevent additional cases. A systematic review of travel-associated LD events (cases, clusters, outbreaks) and of environmental studies of Legionella contamination in accommodation sites was conducted. Two databases were searched (PubMed and EMBASE). Data were extracted from 50 peer-reviewed articles that provided microbiological and epidemiological evidence for linking the accommodation sites with LD. The strength of evidence was classified as strong, possible, and probable. Three of the 21 hotel-associated events identified and four of nine ship-associated events occurred repeatedly on the same site. Of 197 hotel-associated cases, 158 (80.2%) were linked to hotel cooling towers and/or potable water systems. Ship-associated cases were most commonly linked to hot tubs (59/83, 71.1%). Common contributing factors included inadequate disinfection, maintenance, and monitoring; water stagnation; poor temperature control; and poor ventilation. Across all 30 events, Legionella concentrations in suspected water sources were >10,000 cfu/L, <10,000 cfu/L, and unknown in 11, 3, and 13 events, respectively. In five events, Legionella was not detected only after repeated disinfections. In environmental studies, Legionella was detected in 81.1% of ferries (23/28) and 48.9% of hotels (587/1,200), while all 12 cruise ships examined were negative. This review highlights the need for LD awareness strategies targeting operators of accommodation sites. Increased standardization of LD investigation and reporting, and more rigorous follow-up of LD events, would help generate stronger, more comparable evidence on LD sources, contributing factors, and control measure effectiveness.


Prevalence of Psychiatric Illness among Residents of Old Age Homes in Northern India.
There are many factors which compelled older adults to live in old age homes (OAHs) and vulnerable to psychological problems. Studies reported high prevalence of mental health problems (20%-60%) among elderlies of OAHs. Therefore, the study was conducted to explore prevalence of psychiatric illness (PI) among residents of OAHs of Northern India. The present study was conducted in OAHs of Districts Bareilly, Lucknow, Varanasi, Dehradun, and Haridwar, using cross-sectional descriptive study method. Sample Size were 306 (male - 98 [32.5%] and female n = 208 [68%]) residing in OAHs selected by means of purposive sampling. Inclusion criteria: (a) older adults aged 60 years and above residing in OAHs and able to communicate. (b) Staying in OAHs for 6 months or more. (c) Able to understand comprehends and reply to questions and (d) Giving written informed consent. Exclusion criteria: (a) Residents who declined/not interested to participate in the study. (b) Residents having any sensory impairment/physical health problem which can impede the interview. Research tools were (i) a semi-structured pro forma, (ii) Hindi Mental Status Examination, (iii) Survey psychiatric assessment schedule, and (iv) Schedules for clinical assessment in neuropsychiatry-based clinical interview for diagnosis of PIs according to International Classification of Disease 10. The quantitative data obtained was analyzed by means of frequency tables. The results show overall prevalence of PI is 43% among residents of OAHs. The prevalence of PI was found to be higher among females compared to males. Depression was the most common among the residents of OAHs. There is an urgent need of trained professionals to provide professional help for highly prevalent psychiatric disorders among residents of OAHs.


Differential magnitude coding of gains and omitted rewards in the ventral striatum.
Physiologic studies revealed that neurons in the dopaminergic midbrain of non-human primates encode reward prediction errors. It was furthermore shown that reward prediction errors are adaptively scaled with respect to the range of possible outcomes, enabling sensitive encoding for a large range of reward values. Congruently, neuroimaging studies in humans demonstrated that BOLD-responses in the ventral striatum encode reward prediction errors in similar fashion as dopaminergic midbrain neurons, suggesting that these BOLD-responses may be driven by dopaminergic midbrain activity. However, neuroimaging results are ambiguous with respect to the adaptive scaling of reward prediction errors, leading to the conjecture that under certain circumstances other than dopaminergic midbrain input may drive ventral striatal BOLD-responses. The goal of this study was to substantiate whether BOLD-responses in the ventral striatum rather respond to adaptively scaled reward prediction errors or absolute reward magnitude. In addition, we aimed to identify neuronal structures modulating activity in the ventral striatum. Sixteen healthy participants played a wheel of fortune game, where they could win three differently valued rewards while being scanned. BOLD-responses increased after gaining rewards; this gain was however independent of the absolute reward magnitude. In contrast BOLD-responses upon reward omission decreased with reward magnitude. A psychophysiological interaction analysis identified a cluster in the brainstem in proximity of the dorsal raphe nucleus, a cluster in the lateral orbitofrontal cortex, and a cluster in the rostral cingulate zone. These clusters changed their connectivity with the ventral striatum in relation to the absolute reward magnitude in reward omission trials.


WHO guidelines for the programmatic management of drug-resistant tuberculosis: 2011 update.
The production of guidelines for the management of drug-resistant tuberculosis (TB) fits the mandate of the World Health Organization (WHO) to support countries in the reinforcement of patient care. WHO commissioned external reviews to summarise evidence on priority questions regarding case-finding, treatment regimens for multidrug-resistant TB (MDR-TB), monitoring the response to MDR-TB treatment, and models of care. A multidisciplinary expert panel used the Grading of Recommendations Assessment, Development and Evaluation (GRADE) approach to develop recommendations. The recommendations support the wider use of rapid drug susceptibility testing for isoniazid and rifampicin or rifampicin alone using molecular techniques. Monitoring by sputum culture is important for early detection of failure during treatment. Regimens lasting ≥ 20 months and containing pyrazinamide, a fluoroquinolone, a second-line injectable drug, ethionamide (or prothionamide), and either cycloserine or p-aminosalicylic acid are recommended. The guidelines promote the early use of antiretroviral agents for TB patients with HIV on second-line drug regimens. Systems that primarily employ ambulatory models of care are recommended over others based mainly on hospitalisation. Scientific and medical associations should promote the recommendations among practitioners and public health decision makers involved in MDR-TB care. Controlled trials are needed to improve the quality of existing evidence, particularly on the optimal composition and duration of MDR-TB treatment regimens.


Cancer stem cells as mediators of treatment resistance in brain tumors: status and controversies.
Malignant primary brain tumors are characterized by a short median survival and an almost 100% tumor-related mortality. Despite the addition of new chemotherapy regimes, the overall survival has improved marginally, and radiotherapy is only transiently effective, illustrating the profound impact of treatment resistance on prognosis. Recent studies suggest that a small subpopulation of cancer stem cells (CSCs) has the capacity to repopulate tumors and drive malignant progression and mediate radio- and chemoresistance. This implies that future therapies should turn from the elimination of the rapidly dividing, but differentiated tumor cells, to specifically targeting the minority of tumor cells that repopulate the tumor. Although there exists some support for the CSC hypothesis, there remain many uncertainties regarding theoretical, technical, and interpretational aspects of the data supporting it. If correct, the CSC hypothesis could have profound implications for the way tumors are classified and treated. In this review of the literature, we provide original data and hypotheses supporting alternative explanations and outline some of the therapeutic implications that can be derived.


LRIG1 is a novel negative regulator of the Met receptor and opposes Met and Her2 synergy.
The Met receptor tyrosine kinase regulates a complex array of cellular behaviors collectively known as "invasive growth." While essential for normal development and wound repair, this program is frequently co-opted by tumors to promote their own growth, motility, and invasion. Met is overexpressed in a variety of human tumors, and this aberrant expression correlates with poor patient prognosis. Previous studies indicate that Met receptor levels are governed in part by cbl-mediated ubiquitination and degradation, and uncoupling of Met from cbl-mediated ubiquitination promotes its transforming activity. Here we describe a novel mechanism for Met degradation. We find that the Met receptor interacts with the transmembrane protein LRIG1 independent of hepatocyte growth factor (HGF) stimulation and that LRIG1 destabilizes the Met receptor in a cbl-independent manner. Overexpression of LRIG1 destabilizes endogenous Met receptor in breast cancer cells and impairs their ability to respond to HGF. LRIG1 knockdown increases Met receptor half-life, indicating that it plays an essential role in Met degradation. Finally, LRIG1 opposes Met synergy with the ErbB2/Her2 receptor tyrosine kinase in driving cellular invasion. We conclude that LRIG1 is a novel suppressor of Met function, serving to regulate cellular receptor levels by promoting Met degradation in a ligand- and cbl-independent manner.


Intervisit Repeatability of Retinal Blood Oximetry and Total Retinal Blood Flow Under Varying Systemic Blood Gas Oxygen Saturations.
The aim of the study is to validate and calibrate the Doppler spectral-domain optical coherence tomography (SD-OCT) derived total retinal blood flow (TRBF) and metabolic hyperspectral retinal camera (MHRC) derived oxygen saturation (SO2) of major retinal vessels in human volunteers using a novel and exact provocation technique (RespirAct) that allows the precise control of the end-tidal partial pressure of oxygen (PETO2). Between visit repeatability of the TRBF and retinal blood SO2 also were studied. One eye of 11 young healthy subjects was chosen randomly for the study. Total retinal blood flow and retinal SO2 measurements were obtained under conditions of normoxia, hyperoxia, and hypoxia. The order of hyperoxia and hypoxia was randomized between subjects. The measurements were repeated after a week interval. When the arterial PETO2 was increased from baseline (PETO2 = 100 mm Hg) to 200 and 300 mm Hg, the TRBF significantly reduced (P = 0.02) from 44.60 (±8.9) to 40.28 (±8.9) and 36.23 (±4.6) μL/min. Lowering the arterial PETO2, from baseline to 80, 60, and 50 mm Hg, TRBF significantly increased (P = 0.04) from 43.17 (±12.7) to 45.19 (±5.5), 49.71 (±13.4), and 52.89 (±10.9) μL/min with simultaneous reduction in the arterial blood SO2 content from 99.3% (±5.8) to 95.6% (±5.1), 89.6% (±2.8), and 83.3% (±3.9), respectively (P = 0.00). The coefficient of repeatability (COR) of TRBF, retinal arterial, and venous blood SO2 values are 21.8 μL/min, 18.4%, and 15.2%, respectively. Total retinal blood flow and retinal blood SO2 measurements performed under safe levels of hypoxia and hyperoxia were repeatable in healthy adults over the range of SO2 investigated.


Approach to thyroid nodules in children and adolescents.
Thyroid nodule prevalence is about 1.8% in healthy children; however, malignancy frequency is higher than in adults. Approximately 26.4% of thyroid nodules generate thyroid cancer in childhood. Coexisting thyroid disease, history of irradiation of the neck, post-pubertal age, female sex, and thyroid malignancy in the family are risk factors for developing nodules. After evaluation of the medical history and detailed physical examination, the second step is assessment of thyroid function and measurement of calcitonin level. Thyroid stimulating hormone (TSH) value in the upper range seems to be correlated with cancer. Calcitonin levels must be evaluated, especially if medullary cancer is suspected. Ultrasonography (USG) is the first-line imaging tool in the diagnosis of thyroid nodules. It gives information about the nodule size, echogenicity and location. Hypoechogenicity, microcalcifications, undefined margins, high internodular vascular flow, and subcapsular localization are clues of malignant lesions. Scintigraphy is only recommended in a solid nodule with the presence of suppressed TSH. Fine-needle aspiration biopsy (FNAB) has 90% accuracy and is very useful in the selection of patients for surgery. It must be applied to all nodules ≥1 cm and nodules ≤1 cm suspicious for malignancy. The other diagnostic tools are elastography, immunocytochemical markers and genetic evaluation. In the management of thyroid nodules, surgery is advised, especially if there is difficulty in distinguishing benign lesions from carcinoma.


Stress axis regulation during social ascension in a group-living cichlid fish.
Animals living in groups often form social hierarchies, with characteristic behaviours and physiologies associated with rank. However, when social opportunities arise and a subordinate ascends into a dominant position, quick adjustments are necessary to secure this position. Such periods of social transition are typically associated with elevated glucocorticoid production, but the precise regulation of the stress axis during these occasions is not well understood. Using the group-living cichlid, Neolamprologus pulcher, the effects of social ascension on the stress axis were assessed. Ascenders rapidly filled experimentally created vacancies, adopting a dominant behavioural phenotype within 72 h-elevating aggression, activity, and workload, while receiving high rates of affiliative behaviours from their group members. Despite assuming behavioural dominance within their groups, ascenders displayed higher cortisol levels than dominants three days post-ascension. Additionally, compared to subordinates, ascenders had increased transcript abundance of steroidogenic acute regulatory protein (star) and cytochrome p450 side-chain cleavage enzyme (p450scc) in the head kidney, indicating activation of the stress axis. Cortisol levels were lowest in ascenders that displayed low rates of aggression, potentially reflecting the reestablishment of social stability in these groups. Increased transcript abundance of both glucocorticoid receptors (gr1 and gr2) in the brain's preoptic area (POA) of ascenders compared to dominants suggested an enhanced capacity for cortisol regulation via negative feedback. Our results reveal a regulatory cascade of behavioural and physiological interactions and highlight the importance of investigating the underlying mechanisms regulating the stress axis.


Cortical dynein controls microtubule dynamics to generate pulling forces that position microtubule asters.
Dynein at the cortex contributes to microtubule-based positioning processes such as spindle positioning during embryonic cell division and centrosome positioning during fibroblast migration. To investigate how cortical dynein interacts with microtubule ends to generate force and how this functional association impacts positioning, we have reconstituted the 'cortical' interaction between dynein and dynamic microtubule ends in an in vitro system using microfabricated barriers. We show that barrier-attached dynein captures microtubule ends, inhibits growth, and triggers microtubule catastrophes, thereby controlling microtubule length. The subsequent interaction with shrinking microtubule ends generates pulling forces up to several pN. By combining experiments in microchambers with a theoretical description of aster mechanics, we show that dynein-mediated pulling forces lead to the reliable centering of microtubule asters in simple confining geometries. Our results demonstrate the intrinsic ability of cortical microtubule-dynein interactions to regulate microtubule dynamics and drive positioning processes in living cells.


Role of the accident and emergency department in the non-heart-beating donor programme in Leicester.
To describe the development of a non-heart-beating donor (NHBD) programme in an accident and emergency (A&E) department over a three year period. The A&E department at the Leicester Royal Infirmary at present deals with approximately 200 prehospital cardiopulmonary arrests per year. A programme of kidney retrieval from non-heart-beating donors was started in April 1992. Strict criteria for admission to the programme, appropriate consent procedures, facilities, lines of communication, and feedback were developed to enhance its success. Of 66 patients referred to the NHBD programme over a three year period from 1 April 1992, 51 sets of relatives were available to be asked for possible organ donation, and 34 sets (66%) gave their consent. Twenty five patients had successful in situ perfusion of the kidneys. Forty seven organs were retrieved and 34 went on to be transplanted. To date, 27 kidneys are still working. As a result, 23.8% of kidneys transplanted in Leicester over this time period have been from the NHBD programme. The NHBD programme in Leicester has proved very successful, requiring organisation of resources and personnel both from the transplant service and the A&E department. The programme has provided such a significant boost to the renal transplant rate in Leicester that other hospitals with large A&E departments should consider setting up similar programmes.


Quality of life and supported employment: a case study of three women with developmental disabilities.
This paper presents the findings of a qualitative case study of the quality of life of three women with developmental disabilities. In-depth interview data were collected from the women who had previously worked in a sheltered workshop environment and were, at the time of study, working in supported employment in the community. Interviews were also conducted with each woman's job coach and a family member or close friend. On-site observations of the women in their supported employment environments were also undertaken. Results show that the women's own perceptions about their quality of life were not always as family members and job coaches expected, and that the purported benefits of supported employment for the three women with developmental disabilities were not always realized. Occupational therapists have an opportunity to facilitate the occupational performance of adults with developmental disabilities but they must be willing to engage in wider health and social policy change if they are to make significant gains.


The transcriptome profile of the mosquito Culex quinquefasciatus following permethrin selection.
To gain valuable insights into the gene interaction and the complex regulation system involved in the development of insecticide resistance in mosquitoes Culex quinquefasciatus, we conducted a whole transcriptome analysis of Culex mosquitoes following permethrin selection. Gene expression profiles for the lower resistant parental mosquito strain HAmCq(G0) and their permethrin-selected high resistant offspring HAmCq(G8) were compared and a total of 367 and 3982 genes were found to be up- and down-regulated, respectively, in HAmCq(G8), indicating that multiple genes are involved in response to permethrin selection. However, a similar overall cumulative gene expression abundance was identified between up- and down-regulated genes in HAmCq(G8) mosquitoes following permethrin selection, suggesting a homeostatic response to insecticides through a balancing of the up- and down-regulation of the genes. While structural and/or cuticular structural functions were the only two enriched GO terms for down-regulated genes, the enriched GO terms obtained for the up-regulated genes occurred primarily among the catalytic and metabolic functions where they represented three functional categories: electron carrier activity, binding, and catalytic activity. Interestingly, the functional GO terms in these three functional categories were overwhelmingly overrepresented in P450s and proteases/serine proteases. The important role played by P450s in the development of insecticide resistance has been extensively studied but the function of proteases/serine proteases in resistance is less well understood. Hence, the characterization of the functions of these proteins, including their digestive, catalytic and proteinase activities; regulation of signaling transduction and protein trafficking, immunity and storage; and their precise function in the development of insecticide resistance in mosquitoes will provide new insights into how genes are interconnected and regulated in resistance.


Evaluation of the suitability of weekly peak expiratory flow rate measurements in monitoring annual decline in lung function among patients with asthma and chronic bronchitis.
Early detection and treatment of patients with asthma or chronic bronchitis who have a rapid annual decline in lung function is essential in order to improve their long-term prognosis. This annual rate of decline can be assessed accurately by monitoring the forced expiratory volume in one second (FEV1) which is a routine procedure in hospital respiratory laboratories but not in general practice. General practitioners usually measure patients' peak expiratory flow rate (peak flow) to evaluate lung function. If annual decline in lung function can be assessed by monitoring peak flow, this method could be used in general practice for detecting patients at an early stage who have a rapid decline. A study aimed to investigate the long-term correlation between FEV1 and peak flow among a group of patients in Nijmegen, the Netherlands. FEV1 and peak flow were monitored in 53 patients with moderate asthma and 78 patients with moderate chronic bronchitis over four years. FEV1 was measured in a laboratory once every six months and peak flow was measured by patients once a week. The correlation between the two sets of measurements was studied for each patient. Four-year data for 83 of the 131 patients were analysed; the other 48 patients received inhaled steroids during the second half of the study period so their data were not considered for all the analyses. Of the 83 patients, 35 (42%) showed a decrease in both FEV1 and peak flow. Thirty six patients (43%) showed a decrease in FEV1 and an increase in peak flow. Four patients (5%) showed an increase in FEV1 and a decrease in peak flow and eight patients (10%) showed an increase in both rates. Approximately similar results were seen in a separate analysis of all 131 patients during the first two years of the study. No long-term correlation was found between FEV1 and peak flow. Peak flow is not capable of detecting annual decline in lung function. Therefore it cannot be used to detect patients with asthma or chronic bronchitis who have a rapid annual decline in lung function. Spirometers, which measure peak flow and FEV1, could be used in general practice. These would allow general practitioners to continue measuring peak flow in order to assess short-term changes in lung function while providing an important means for monitoring FEV1 to assess long-term changes in lung function.


In vivo 13C NMR studies of compartmentalized cerebral carbohydrate metabolism.
Localized 13C nuclear magnetic resonance (NMR) spectroscopy provides a unique window for studying cerebral carbohydrate metabolism through, e.g. the completely non-invasive measurement of cerebral glucose and glycogen metabolism. In addition, label incorporation into amino acid neurotransmitters such as glutamate (Glu), GABA and aspartate can be measured providing information on Krebs cycle flux and oxidative metabolism. Given the compartmentation of key enzymes such as pyruvate carboxylase and glutamine synthetase, the detection of label incorporation into glutamine indicated that neuronal and glial metabolism can be measured in vivo. The purpose of this paper is to provide a critical overview of these recent advances into measuring compartmentation of brain energy metabolism using localized in vivo 13C NMR spectroscopy. The studies reviewed herein showed that anaplerosis is significant in brain, as is oxidative ATP generation in glia and the rate of glial glutamine synthesis attributed to the replenishment of the neuronal Glu pool and that brain glycogen metabolism is slow under resting conditions. This new modality promises to provide a new investigative tool to study aspects of normal and diseased brain hitherto unaccessible, such as the interplay between glutamatergic action, glucose and glycogen metabolism during brain activation, and the derangements thereof in patients with hepatic encephalopathy, neurodegenerative diseases and diabetes.


Comparison of plasma lipids and vitamin E in young and middle-aged subjects on potato crisps fried in palmolein and highly oleic sunflower oil.
We previously found no difference in healthy young adults' plasma cholesterols between palmolein and olive oil as the major dietary lipid, although the former is high in palmitic acid (16:0) but the latter in oleic acid (18:1 cis). In the experiment reported here we compared the effects of palmolein against another monounsaturated oil, highly oleic sunflower oil (HOSO), on plasma cholesterol in both young and middle-aged healthy adults. The test oils were provided as frying oil of potato crisps (150 g/day in men; 100 g/day in women) against low-fat background diets in free-living motivated volunteers. The design was a randomised double-blind 4-week/3-week crossover trial. Compliance was monitored with continuous dietary diaries and by measuring (fasting) plasma lipid fatty-acid pattern. Plasma lipids and vitamin-E compounds were measured at the start and twice at the end of each test period. In combined young plus older subjects (n = 42) mean plasma total and low-density-lipoprotein cholesterol (LDL-c) values were both 7% (significantly) lower on HOSO than on palmolein, but because high-density-lipoprotein cholesterol (HDL-c) was also 5% lower, the LDL-c/HDL-c ratio was only 3% lower on HOSO than on palmolein. The difference between the present results with HOSO and previous results with olive oil both compared against palmolein suggest that olive oil is associated with higher plasma cholesterols than other monounsaturated oils. In both the young and older subgroup, LDL-c was lower on HOSO but because HDL-c moved down too in the young subgroup, the LDL-c/HDL-c ratio was lower on HOSO only in the older subjects. Palmolein has an unusual pattern of E vitamins, with a high content of tocotrienols, notably the gamma-isomer. Unlike alpha-tocopherol however, there was no sign of these tocotrienols in subjects' plasmas.


Optical lateral inhibition networks that use self-linearized self-electro-optic-effect devices: theory and experiment.
The self-linearized self-electro-optic-effect-device (SL-SEED) phenomenon observed with a quantum-well modulator and photodiode serial combination is one of the few practical routes to optical subtraction. A family of optical lateral inhibition architectures based on the SL-SEED that incorporate optical feedback is introduced and their operation confirmed in simulation. A successful experimental demonstration based on these ideas, performing edge-contrast enhancement by lateral inhibition, is described. System interconnections are both optical and electrical, with nonlocal interconnections being made optically by the use of diffractive elements.


Assessing somatization in urologic chronic pelvic pain syndrome.
This study examined the prevalence of somatization disorder in Urological Chronic Pelvic Pain Syndrome (UCPPS) and the utility of two self-report symptom screening tools for assessment of somatization in patients with UCPPS. The study sample included 65 patients with UCPPS who enrolled in the Multidisciplinary Approach to the Study of Chronic Pelvic Pain (MAPP) Study at Washington University. Patients completed the PolySymptomatic PolySyndromic Questionnaire (PSPS-Q) (n = 64) and the Patient Health Questionnaire-15 Somatic Symptom Severity Scale (PHQ-15) (n = 50). Review of patient medical records found that only 47% (n = 30) contained sufficient documentation to assess Perley-Guze criteria for somatization disorder. Few (only 6.5%) of the UCPPS sample met Perley-Guze criteria for definite somatization disorder. Perley-Guze somatization disorder was predicted by definite PSPS-Q somatization with at least 75% sensitivity and specificity. Perley-Guze somatization disorder was predicted by severe (> 15) PHQ-15 threshold that had > 90% sensitivity and specificity but was met by only 16% of patients. The moderate (> 10) PHQ-15 threshold had higher sensitivity (100%) but lower specificity (52%) and was met by 52% of the sample. The PHQ-15 is brief, but it measures symptoms constituting only one dimension of somatization. The PSPS-Q uniquely captures two conceptual dimensions inherent in the definition of somatization disorder, both number of symptoms and symptom distribution across multiple organ systems, with relevance for UCPPS as a syndrome that is not just a collection of urological symptoms but a broader syndrome with symptoms extending beyond the urological system.


In vitro and in vivo imaging of initial B-T-cell interactions in the setting of B-cell based cancer immunotherapy.
There has been a growing interest in the use of B cells for cancer vaccines, since they have yielded promising results in preclinical animal models. Contrary to dendritic cells (DCs), we know little about the migration behavior of B cells in vivo. Therefore, we investigated the interactions between CD40-activated B (CD40B) cells and cytotoxic T cells in vitro and the migration behavior of CD40B cells in vivo. Dynamic interactions of human antigen-presenting cells (APCs) and T cells were observed by time-lapse video microscopy. The migratory and chemoattractant potential of CD40B cells was analyzed in vitro and in vivo using flow cytometry, standard transwell migration assays, and imaging of fluorescently labeled murine CD40B cells. Murine CD40B cells show migratory features similar to human CD40B cells. They express important lymph node homing receptors which were functional and induced chemotaxis of T cells in vitro. Striking differences were observed with regard to interactions of human APCs with T cells. CD40B cells differ from DCs by displaying a rapid migratory pattern undergoing highly dynamic, short-lived and sequential interactions with T cells. In vivo, CD40B cells are home to the secondary lymphoid organs where they accumulate in the B cell zone before traveling to the B/T cell boundary. Moreover, intravenous (i.v.) administration of murine CD40B cells induced an antigen-specific cytotoxic T cell response. Taken together, this data show that CD40B cells home secondary lymphoid organs where they physically interact with T cells to induce antigen-specific T cell responses, thus underscoring their potential as cellular adjuvant for cancer immunotherapy.


Prophylactic hypogastric artery ballooning in a patient with complete placenta previa and increta.
Abnormal attachment of the placenta (Placenta accreta, increta, and percreta) is an uncommon but potentially lethal cause of maternal mortality from massive postpartum hemorrhage. A 33-yr-old woman, who had been diagnosed with a placenta previa, was referred at 30 weeks gestation. On ultrasound, a complete type of placenta previa and multiple intraplacental lacunae, suggestive of placenta accreta, were noted. For further evaluation of the placenta, pelvis MRI was performed and revealed findings suspicious of a placenta increta. An elective cesarean delivery and subsequent hysterectomy were planned for the patient at 38 weeks gestation. On the day of delivery, endovascular catheters for balloon occlusion were placed within the hypogastric arteries, prior to the cesarean section. In the operating room, immediately after the delivery of the baby, bilateral hypogastric arteries were occluded by inflation of the balloons in the catheters previously placed within. With the placenta retained within the uterus, a total hysterectomy was performed in the usual fashion. The occluding balloons were deflated after closure of the vaginal cuff with hemostasis. The patient had stable vital signs and normal laboratory findings during the recovery period; she was discharged six days after delivery without complications. The final pathology confirmed a placenta increta.


Potential benefits of phytochemicals against Alzheimer's disease.
Our current therapeutic drugs for Alzheimer's disease are predominantly derived from the alkaloid class of plant phytochemicals. These drugs, such as galantamine and rivastigmine, attenuate the decline in the cholinergic system but, as the alkaloids occupy the most dangerous end of the phytochemical spectrum (indeed they function as feeding deterrents and poisons to other organisms within the plant itself), they are often associated with unpleasant side effects. In addition, these cholinesterase inhibiting alkaloids target only one system in a disorder, which is typified by multifactorial deficits. The present paper will look at the more benign terpene (such as Ginkgo biloba, Ginseng, Melissa officinalis (lemon balm) and Salvia lavandulaefolia (sage)) and phenolic (such as resveratrol) phytochemicals; arguing that they offer a safer alternative and that, as well as demonstrating efficacy in cholinesterase inhibition, these phytochemicals are able to target other salient systems such as cerebral blood flow, free radical scavenging, anti-inflammation, inhibition of amyloid-β neurotoxicity, glucoregulation and interaction with other neurotransmitters (such as γ-aminobutyric acid) and signalling pathways (e.g. via kinase enzymes).


The Present and Future of Clinical Management in Metastatic Breast Cancer.
Regardless of the advances in our ability to detect early and treat breast cancer, it is still one of the common types of malignancy worldwide, with the majority of patients decease upon metastatic disease. Nevertheless, due to these advances, we have extensively characterized the drivers and molecular profiling of breast cancer and further dividing it into subtypes. These subgroups are based on immunohistological markers (Estrogen Receptor-ER; Progesterone Receptor-PR and Human Epidermal Growth Factor Receptor 2-HER-2) and transcriptomic signatures with distinct therapeutic approaches and regiments. These therapeutic approaches include targeted therapy (HER-2+), endocrine therapy (HR+) or chemotherapy (TNBC) with optional combination radiotherapy, depending on clinical stage. Technological and scientific advances in the identification of molecular pathways that contribute to therapy-resistance and establishment of metastatic disease, have provided the rationale for revolutionary targeted approaches against Cyclin-Dependent Kinases 4/6 (CDK4/6), PI3 Kinase (PI3K), Poly ADP Ribose Polymerase (PARP) and Programmed Death-Ligand 1 (PD-L1), among others. In this review, we focus on the comprehensive overview of epidemiology and current standard of care treatment of metastatic breast cancer, along with ongoing clinical trials. Towards this goal, we utilized available literature from PubMed and ongoing clinical trial information from clinicaltrials.gov to reflect the up to date and future treatment options for metastatic breast cancer.


Blood-Based Oligomeric and Other Protein Variant Biomarkers to Facilitate Pre-Symptomatic Diagnosis and Staging of Alzheimer's Disease.
Oligomeric forms of amyloid-β (Aβ), tau, and TDP-43 play important roles in Alzheimer's disease (AD), and therefore are promising biomarkers. We previously generated single chain antibody fragments (scFvs) that selectively bind disease-related variants of these proteins including A4, C6T, and E1, which bind different oligomeric Aβ variants; D11C, which binds oligomeric tau; and AD-TDP1 and AD-TDP2, which bind disease related TDP-43 variants. To determine the utility of these disease-related variants as early biomarkers, we first analyzed 11 human sera samples obtained ∼2 years prior to an initial mild cognitive impairment (MCI) diagnosis. While the subsequent diagnoses for the cases covered several different conditions, all samples had elevated protein variant levels relative to the plasma controls although with different individual biomarker profiles. We then analyzed a set of longitudinal human plasma samples from four AD (encompassing time points prior to MCI diagnosis and continuing until after conversion to AD) and two control cases. Pre-MCI samples were characterized by high TDP-43 variant levels, MCI samples by high Aβ variant levels, and AD samples by high Aβ and tau variant levels. Sample time points ranged from ∼7 years pre-MCI to ∼9 years after AD conversion. Bivariate correlations showed a negative correlation with TDP-43 levels and positive correlations with cumulative Aβ and oligomeric tau levels indicating an increase in neurodegenerative processes with time in AD. Detection of disease related protein variants not only readily selects AD cases from controls, but also stages progression of AD and holds promise for a pre-symptomatic blood-based biomarker profile for AD.


Disrupting Crystal Growth through Molecular Recognition: Designer Therapies for Kidney Stone Prevention.
Aberrant crystallization within the human body can lead to several disease states or adverse outcomes, yet much remains to be understood about the critical stages leading to these events, which can include crystal nucleation and growth, crystal aggregation, and the adhesion of crystals to cells. Kidney stones, which are aggregates of single crystals with physiological origins, are particularly illustrative of pathological crystallization, with 10% of the U.S. population experiencing at least one stone occurrence in their lifetimes. The human record of kidney stones is more than 2000 years old, as noted by Hippocrates in his renowned oath and much later by Robert Hooke in his treatise Micrographia. William Hyde Wollaston, who was a physician, chemist, physicist, and crystallographer, was fascinated with stones, leading him to discover an unusual stone that he described in 1810 as cystic oxide, later corrected to cystine. Despite this long history, however, a fundamental understanding of the stages of stone formation and the rational design of therapies for stone prevention have remained elusive.This Account reviews discoveries and advances from our laboratories that have unraveled the complex crystal growth mechanisms of l-cystine, which forms l-cystine kidney stones in at least 20 000 individuals in the U.S. alone. Although l-cystine stones affect fewer individuals than common calcium oxalate stones, they are usually larger, recur more frequently, and are more likely to cause chronic kidney disease. Real-time in situ atomic force microscopy (AFM) reveals that the crystal growth of hexagonal l-cystine is characterized by a complex mechanism in which six interlaced anisotropic spirals grow synchronously, emanating from a single screw dislocation to generate a micromorphology with the appearance of stacked hexagonal islands. In contrast, proximal heterochiral dislocations produce features that appear to be spirals but actually are closed loops, akin to a Frank-Read source. These unusual and aesthetic growth patterns can be explained by the coincidence of the dislocation Burgers vector and the crystallographic 61 screw axis. Inhibiting l-cystine crystal growth is key to preventing stone formation. Decades of studies of "tailor-made additives", which are imposter molecules that closely resemble the solute and bind to crystal faces through molecular recognition, have demonstrated their effects on crystal properties such as morphology and polymorphism. The ability to visualize crystal growth in real time by AFM enables quantitative measurements of step velocities and, by extension, the effect of prospective inhibitors on growth rates, which can then be used to deduce inhibition mechanisms. Investigations with a wide range of prospective inhibitors revealed the importance of precise molecular recognition for binding l-cystine imposters to crystal sites, which results in step pinning and the inhibition of step advancement as well as the growth of bulk crystals. Moreover, select inhibitors of crystal growth, measured in vitro, reduce or eliminate stone formation in knockout mouse models of cystinuria, promising a new pathway to l-cystine stone prevention. These observations have wide-ranging implications for the design of therapies based on tailor-made additives for diseases associated with aberrant crystallization, from disease-related stones to "xenostones" that form in vivo because of the crystallization of low-solubility therapeutic agents such as antiretroviral agents.


Some coumarins and benzoxazinones as potent paraoxonase 1 inhibitors.
In this study, we aimed to investigate the effect of some coumarin and benzoxazinone derivatives on the activity of human PON1. Human serum paraoxonase 1 was purified from fresh human serum blood by two-step procedures that are ammonium sulfate precipitation (60-80%) and then hydrophobic interaction chromatography (Sepharose 4B, L-tyrosine and 1-napthylamine). The enzyme was purified 232-fold with a final specific activity of 27.1 U/mg. In vitro effects of some previously synthesized ionic coumarin or benzoxazinone derivatives (1-21) on purified PON1 activity were investigated. Compound 14 (1-(2,3,4,5,6)-pentamethylbenzyl-3-(6,8-dimethyl-2H-chromen-2-one-4-yl))benzimidazolium chloride was found out as the strongest inhibitor (IC50 = 7.84 μM) for PON1 among the compounds. Kinetic investigation and molecular docking study were evaluated for one of the most active compounds (compound 12) and obtained data showed that this compound is competitive inhibitor of PON1 and interact with Leu262 and Ser263 in the active site of PON1. Moreover, coumarin derivatives were found out as the more potent inhibitors for PON1 than benzoxazinone derivatives.


Improving the permeability of lyophilized collagen-hydroxyapatite scaffolds for cell-based bone regeneration with a gelatin porogen.
Bone tissue engineering using biomaterial scaffolds and culture-expanded osteoprogenitor cells has been demonstrated in several studies; however, it is not yet a clinical reality. One challenge is the optimal design of scaffolds for cell delivery and the identification of scaffold parameters that can delineate success and failure in vivo. Motivated by a previous experiment in which a batch of lyophilized collagen-hydroxyapatite (HA) scaffolds displayed modest bone formation in vivo, despite having large pores and high porosity, we began to investigate the effect of scaffold permeability on bone formation. Herein, we fabricated scaffolds with a permeability of 2.17 ± 1.63 × 10-9 m4 /(N s) and fourfold higher using a sacrificial gelatin porogen. Scaffolds were seeded with mouse bone marrow stromal cells carrying a fluorescent reporter for osteoblast differentiation and implanted into critical-size calvarial defects in immunodeficient mice. The porogen scaffold group containing a 1:1 ratio of solids to beads was significantly more radiopaque than the scaffold group without the bead porogen 3 weeks after implantation. Quantitative histomorphometry uncovered the same trend between the 1:1 group and scaffolds without porogen found in the radiographic data; however, this was not statistically significant here. Taken together, the X-ray and histology suggest that the 1:1 ratio of porogen to scaffold solids, resulting in a fourfold increase in permeability, may enhance bone formation when compared to scaffolds without porogen. Scaffold permeability can be a useful quality control measure before implantation and this practice should improve the consistency and efficacy of cell-based bone tissue engineering. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 1580-1590, 2016.


Measuring and managing the work environment of the mid-level provider--the neglected human resource.
Much has been written in the past decade about the health workforce crisis that is crippling health service delivery in many middle-income and low-income countries. Countries having lost most of their highly qualified health care professionals to migration increasingly rely on mid-level providers as the mainstay for health services delivery. Mid-level providers are health workers who perform tasks conventionally associated with more highly trained and internationally mobile workers. Their training usually has lower entry requirements and is for shorter periods (usually two to four years). Our study aimed to explore a neglected but crucial aspect of human resources for health in Africa: the provision of a work environment that will promote motivation and performance of mid-level providers. This paper explores the work environment of mid-level providers in Malawi, and contributes to the validation of an instrument to measure the work environment of mid-level providers in low-income countries. Three districts were purposely sampled from each of the three geographical regions in Malawi. A total of 34 health facilities from the three districts were included in the study. All staff in each of the facilities were included in the sampling frame. A total of 153 staff members consented to be interviewed. Participants completed measures of perceptions of work environment, burnout and job satisfaction. The Healthcare Provider Work Index, derived through Principal Components Analysis and Rasch Analysis of our modification of an existing questionnaire, constituted four subscales, measuring: (1) levels of staffing and resources; (2) management support; (3) workplace relationships; and (4) control over practice. Multivariate analysis indicated that scores on the Work Index significantly predicted key variables concerning motivation and attrition such as emotional exhaustion, job satisfaction, satisfaction with the profession and plans to leave the current post within 12 months. Additionally, the findings show that mid-level medical staff (i.e. clinical officers and medical assistants) are significantly less satisfied than mid-level nurses (i.e. enrolled nurses) with their work environments, particularly their workplace relationships. They also experience significantly greater levels of dissatisfaction with their jobs and with their profession. The Healthcare Provider Work Index identifies factors salient to improving job satisfaction and work performance among mid-level cadres in resource-poor settings. The extent to which these results can be generalized beyond the current sample must be established. The poor motivational environment in which clinical officers and medical assistants work in comparison to that of nurses is of concern, as these staff members are increasingly being asked to take on leadership roles and greater levels of clinical responsibility. More research on mid-level providers is needed, as they are the mainstay of health service delivery in many low-income countries. This paper contributes to a methodology for exploring the work environment of mid-level providers in low-income countries and identifies several areas needing further research.


Heart rate variability during social interactions in children with and without psychopathology: a meta-analysis.
The inability to regulate autonomic activity during social interactions is believed to contribute to social and emotional dysregulation in children. Research has employed heart rate variability (HRV) during both socially engaging and socially disengaging dyadic tasks between children and adults to assess this. We conducted a meta-analysis including evidence from 18 studies comprising 1,544 children who were categorized as either healthy or at risk/diagnosed with psychopathology. Within these groups, we assessed the impact of social engagement and disengagement tasks on HRV. Results showed that social engagement tasks left HRV unaltered to a baseline. Social disengagement, however, was associated with decreases in HRV. In a task that included disengagement and then engagement, HRV was reduced during disengagement but was then restored during the reunion phase (engagement). Children at risk or with a diagnosis for psychopathology, however, failed to show any change in HRV during dyadic social interaction tasks. This was despite a social stressor, the Trier Social Stress Test, causing significantly reduced HRV in both groups. This meta-analysis provides support to suggest HRV may provide a worthwhile context specific marker for the effective regulation of dyadic social interactions in children.


[Determinants of preventive health behavior of the Swiss population. Results of the SOMIPOPS study].
In the context of the first Swiss National Health Survey preventive health behaviour was tested in 3419 Swiss citizens aged over 20 years. A health behaviour index including favourable health attitudes and good utilization of preventive health services was correlated with different social indicators. Age, sex and a good self-rated health were the most important factors associated with a high preventive health behaviour score. Education showed a less strong but also significant influence on positive health behaviour. Overall utilization of health services decreased with high preventive health behavior index.


Treatable abdominal pathologic conditions and unsuspected malignant neoplasms at autopsy in veterans who received mechanical ventilation.
To determine, in medical patients who received mechanical ventilation, the frequency and types of major unexpected diagnoses at autopsy that, if known before death, would probably have led to improved survival (class I errors) or substantively changed management but not survival (class IIB errors). Retrospective cohort study. Six medical intensive care units in a Department of Veterans Affairs Cooperative Study. One hundred seventy-two autopsied patients of the 401 veterans who received mechanical ventilation and died in the hospital. The class I error rate was 12%. Abdominal pathologic conditions--abscesses, bowel perforations, or infarction--were as frequent as pulmonary emboli as a cause of class I errors. While patients with abdominal pathologic conditions generally complained of abdominal pain, results of examination of the abdomen were considered unremarkable in most patients, and the symptom was not pursued. Six percent of patients had extensive malignant neoplasms (class IIB errors). Atypical presentation of potentially treatable abdominal pathologic conditions is a common cause of class I errors in veterans who receive mechanical ventilation. Conversely, several patients with unrecognized terminal conditions underwent intensive intervention. If the information gained at autopsy had been known before death, management would have probably changed substantively in 18% of patients.


Observation of public health risk behaviours, risk communication and hand hygiene at Kansas and Missouri petting zoos--2010-2011.
Outbreaks of human illness have been linked to visiting settings with animal contact throughout developed countries. This study details an observational study of hand hygiene tool availability and recommendations; frequency of risky behaviour; and handwashing attempts by visitors in Kansas (9) and Missouri (4), USA, petting zoos. Handwashing signs and hand hygiene stations were available at the exit of animal-contact areas in 10/13 and 8/13 petting zoos, respectively. Risky behaviours were observed being performed at all petting zoos by at least one visitor. Frequently observed behaviours were as follows: children (10/13 petting zoos) and adults (9/13 petting zoos) touching hands to face within animal-contact areas; animals licking children's and adults' hands (7/13 and 4/13 petting zoos, respectively); and children and adults drinking within animal-contact areas (5/13 petting zoos each). Of 574 visitors observed for hand hygiene when exiting animal-contact areas, 37% (n = 214) of individuals attempted some type of hand hygiene, with male adults, female adults and children attempting at similar rates (32%, 40% and 37%, respectively). Visitors were 4.8× more likely to wash their hands when a staff member was present within or at the exit to the animal-contact area (136/231, 59%) than when no staff member was present (78/343, 23%; P < 0.001, OR = 4.863, 95% CI = 3.380-6.998). Visitors at zoos with a fence as a partial barrier to human-animal contact were 2.3× more likely to wash their hands (188/460, 40.9%) than visitors allowed to enter the animals' yard for contact (26/114, 22.8%; P < 0.001, OR = 2.339, 95% CI = 1.454-3.763). Inconsistencies existed in tool availability, signage and supervision of animal contact. Risk communication was poor, with few petting zoos outlining risks associated with animal contact, or providing recommendations for precautions to be taken to reduce these risks.


Acquired dyslexia in Spanish: a review and some observations on a new case of deep dyslexia.
Readers and writers of Spanish use an orthography that is highly transparent. It has been proposed that readers of Spanish can rely on grapheme-phoneme correspondences, alone, to access meaning or phonology from print. In recent years, a number of case studies have yielded evidence inconsistent with this idea. We review these studies with particular focus on those that report evidence for reading based on direct lexical mappings between print, orthographic representations, and meaning or phonology. We report a new case of acquired literacy impairment in Spanish, MJ, who presents a pattern of preserved abilities and deficits symptomatic of deep dyslexia. The patient is unable to read nonwords, but can read a substantial number of words. Her reading is characterized by the production of semantic, visual, and derivational errors. We argue that MJ has a deficit in her lexical selection ability, common to both her reading and her naming problems. We propose that MJ, and the other cases we review, demonstrate that lexical reading is adopted by skilled readers even in a transparent language.


The pro region is not required for the expression or intracellular routeing of carboxypeptidase E.
Carboxypeptidase E (CPE) is initially synthesized as a larger precursor containing an additional 14-residue propeptide that is highly conserved between human and rat. Previous studies have established that the proenzyme is enzymically active and that deletion of the pro region does not affect the expression of the active enzyme. In the present study the function of the pro region was examined both by deleting this region from CPE and by attaching this region to the N-terminus of albumin. CPE lacking the pro region is sorted into the regulated secretory pathway in AtT-20 cells, based on confocal microscopy and examination of the stimulated secretion of the protein. Stimulation of AtT-20 cells with either forskolin or phorbol 12-myristate 13-acetate induces the secretion of wild-type CPE and of CPE lacking the pro region to similar extents, indicating a similar efficiency of sorting of the mutant. When the pro region of proalbumin is replaced with the pro region of CPE followed by expression in AtT-20 cells, the protein is not sorted into the regulated pathway, based on the lack of stimulated secretion. Confocal microscopy suggests that the proCPE/albumin protein is retained in the endoplasmic reticulum to a greater extent than is proalbumin. Pulse-chase analysis indicates that the pro region of CPE is not efficiently removed from the N-terminus of albumin, and the small amount of propeptide cleavage that does occur takes place soon before secretion of the protein. In contrast, confocal microscopy indicates that the majority of the propeptide is removed from CPE, and that this cleavage occurs in the trans-Golgi network or soon after sorting into the secretory vesicles. Taken together, these results suggest that the pro region of CPE is not required for the expression or intracellular routeing of this protein.


Comparative study of bacterial infection prevalence between cirrhotic patients with and without upper gastrointestinal bleeding.
Bacterial infection is a frequent complication in patients with chronic liver disease, mainly during the advanced stages. There is evidence that the main factors that contribute to a predisposition to infection in cirrhotic patients are related to hepatic failure with consequent immunodeficiency. Invasive procedures (diagnostic or therapeutic) can predispose to bacterial infections, and upper gastrointestinal bleeding (UGB) is considered a potentially important risk factor. A group of cirrhotic patients (child B and C Pugh groups) were evaluated retrospectively by chart reviews regarding the prevalence of bacterial infection during hospitalization to determine whether UGB was a risk factor. An infection was considered present if a specific organ system was identified or if fever (> 38(o)C) persisted for more than 24 hours with associated leukocytosis. Spontaneous bacterial peritonitis was based on classical criteria. Eighty-nine patients were evaluated. Fourty-six patients presented with UGB, and 43 patients had no UGB (control). There were infections recorded in 25/46 (54%) patients with UGB, and 15/43 (35%) in those without UGB (p=0.065). The ratio of the number of infections/admitted patients, was significantly larger in the group with UGB (0.78 +/- 0.89 vs. 0.39 +/- 0.62; p=0.028) since patients had more than one infection. In the UGB group compared to non UGB group, ascites was more frequent (67% vs. 42%; p=0.027); they were more likely to have undergone endoscopic procedures (p<0.001) and the mean +/- SD for platelets count was smaller (96,114 +/- 57,563 vs 145,674 +/- 104,083; p=0.007). The results show that UGB is an important contribution to bacterial infection among Child B and C cirrhotic patients.


Inactivation of endotoxin by human plasma gelsolin.
Septic shock from bacterial endotoxin, triggered by the release of lipopolysaccharide (LPS) molecules from the outer wall of Gram-negative bacteria, is a major cause of human death for which there is no effective treatment once the complex inflammatory pathways stimulated by these small amphipathic molecules are activated. Here we report that plasma gelsolin, a highly conserved human protein, binds LPS from various bacteria with high affinity. Solid-phase binding assays, fluorescence measurements, and functional assays of actin depolymerizing effects show that gelsolin binds more tightly to LPS than it does to its other known lipid ligands, phosphatidylinositol 4,5-bisphosphate and lysophosphatidic acid. Gelsolin also competes with LPS-binding protein (LBP), a high-affinity carrier for LPS. One result of gelsolin-LPS binding is inhibition of the actin binding activity of gelsolin as well as the actin depolymerizing activity of blood serum. Simultaneously, effects of LPS on cellular functions, including cytoskeletal actin remodeling, and collagen-induced platelet activation by pathways independent of toll-like receptors (TLRs) are neutralized by gelsolin and by a peptide based on gelsolin residues 160-169 (GSN160-169) which comprise part of gelsolin's phosphoinositide binding site. Additionally, TLR-dependent NF-kappaB translocation in astrocytes appears to be blocked by gelsolin. These results show a strong effect of LPS on plasma gelsolin function and suggest that some effects of endotoxin in vivo may be mediated or inhibited by plasma gelsolin.


Functional neuroanatomy of human sleep states after zolpidem and placebo: a H215O-PET study.
Changes in the functional organization of the brain during the course of sleep and waking are reflected by different patterns of regional cerebral blood flow (rCBF). To investigate the effect of the hypnotic zolpidem, a benzodiazepine receptor agonist, drug or placebo were administered to eight young, healthy men prior to bedtime. The subjects were sleep-deprived to promote sleep during the 4-h recording period in the positron emission tomography scanner. Intravenous injections of labelled water were administered during pre-drug wakefulness, and during Stage 2, Stage 4 and rapid eye movement (REM) sleep, each injection being followed by an emission scan. Statistical parametric mapping was used to investigate the effects of treatment and sleep states. During sleep (combined Stages 2 and 4, and REM sleep) relative rCBF was lower after zolpidem than after placebo in the basal ganglia and insula, and higher in the parietal cortex. A 'multiple study' analysis of REM sleep revealed that rCBF in the anterior cingulum was lower after zolpidem than after placebo, whereas rCBF in the occipital and parietal cortex, parahippocampal gyrus and cerebellum was higher. When the pooled data (drug and placebo) of Stages 2 and 4 were compared with wakefulness, rCBF was lower in prefrontal cortex and insula, and higher in the occipital and parietal cortex. The results indicate that some differences in rCBF from wakefulness to non-REM sleep are further augmented by zolpidem.


Practical PET Respiratory Motion Correction in Clinical PET/MR.
Respiratory motion during PET acquisition may lead to blurring in resulting images and underestimation of uptake parameters. The advent of integrated PET/MR scanners allows us to exploit the integration of modalities, using high spatial resolution and high-contrast MR images to monitor and correct PET images degraded by motion. We proposed a practical, anatomy-independent MR-based correction strategy for PET data affected by respiratory motion and showed that it can improve image quality both for PET acquired simultaneously to the motion-capturing MR and for PET acquired up to 1 h earlier during a clinical scan. To estimate the respiratory motion, our method needs only an extra 1-min dynamic MR scan, acquired at the end of the clinical PET/MR protocol. A respiratory signal was extracted directly from the PET list-mode data. This signal was used to gate the PET data and to construct a motion model built from the dynamic MR data. The estimated motion was then incorporated into the PET image reconstruction to obtain a single motion-corrected PET image. We evaluated our method in 2 steps. The PET-derived respiratory signal was compared with an MR measure of diaphragmatic displacement via a pencil-beam navigator. The motion-corrected images were compared with uncorrected images with visual inspection, line profiles, and standardized uptake value (SUV) in focally avid lesions. We showed a strong correlation between the PET-derived and MR-derived respiratory signals for 9 patients, with a mean correlation of 0.89. We then showed 4 clinical case study examples ((18)F-FDG and (68)Ga-DOTATATE) using the motion-correction technique, demonstrating improvements in image sharpness and reduction of respiratory artifacts in scans containing pancreatic, liver, and lung lesions as well as cardiac scans. The mean increase in peak SUV (SUV(peak)) and maximum SUV (SUV(max)) in a patient with 4 pancreatic lesions was 23.1% and 34.5% in PET acquired simultaneously with motion-capturing MR, and 17.6% and 24.7% in PET acquired 50 min before as part of the clinical scan. We showed that a respiratory signal can be obtained from raw PET data and that the clinical PET image quality can be improved using only a short additional PET/MR acquisition. Our method does not need external respiratory hardware or modification of the normal clinical MR sequences.


The impact of training and working conditions on junior doctors' intention to leave clinical practice.
The shortage of physicians is an evolving problem throughout the world. In this study we aimed to identify to what extent junior doctors' training and working conditions determine their intention to leave clinical practice after residency training. A prospective cohort study was conducted in 557 junior doctors undergoing residency training in German hospitals. Self-reported specialty training conditions, working conditions and intention to leave clinical practice were measured over three time points. Scales covering training conditions were assessed by structured residency training, professional support, and dealing with lack of knowledge; working conditions were evaluated by work overload, job autonomy and social support, based on the Demand-Control-Support model. Multivariate ordinal logistic regression analyses with random intercept for longitudinal data were applied to determine the odds ratio of having a higher level of intention to leave clinical practice. In the models that considered training and working conditions separately to predict intention to leave clinical practice we found significant baseline effects and change effects. After modelling training and working conditions simultaneously, we found evidence that the change effect of job autonomy (OR 0.77, p = .005) was associated with intention to leave clinical practice, whereas for the training conditions, only the baseline effects of structured residency training (OR 0.74, p = .017) and dealing with lack of knowledge (OR 0.74, p = .026) predicted intention to leave clinical practice. Junior doctors undergoing specialty training experience high workload in hospital practice and intense requirements in terms of specialty training. Our study indicates that simultaneously improving working conditions over time and establishing a high standard of specialty training conditions may prevent junior doctors from considering leaving clinical practice after residency training.


Nonlinear statistical modeling and model discovery for cardiorespiratory data.
We present a Bayesian dynamical inference method for characterizing cardiorespiratory (CR) dynamics in humans by inverse modeling from blood pressure time-series data. The technique is applicable to a broad range of stochastic dynamical models and can be implemented without severe computational demands. A simple nonlinear dynamical model is found that describes a measured blood pressure time series in the primary frequency band of the CR dynamics. The accuracy of the method is investigated using model-generated data with parameters close to the parameters inferred in the experiment. The connection of the inferred model to a well-known beat-to-beat model of the baroreflex is discussed.


Alterations of bone and mineral metabolism in diabetes mellitus. Part I. An overview.
A critical review of the literature leads to the conclusion that alterations of bone and mineral metabolism occur both in diabetic patients and in animals with experimentally induced insulin deficiency syndromes. The coexistence of juvenile insulin-dependent diabetes mellitus (type 1) and radiological evidence of decreased bone mass (osteopenia) appears to be firmly established. Available data support the view that these patients have an increased propensity to skeletal fracture. Adult-onset, non-insulin-dependent diabetic populations, more heterogeneous as regards the type of diabetes, the therapy and the presence of complications or coexistent disease, are characterised by subpopulations with either a decreased, a normal or an increased bone mass. The pathogenesis of diabetic osteopenia is multifactorial. Data obtained from studies employing appropriate animal models of chronic insulin deficiency indicate that various metabolic and hormonal abnormalities may be involved.


Applications of mixture laws for predicting the compositions of tissue phantoms.
This paper generalizes the mixture law methodology for ultrasonic tissue characterization by combining mixture laws used by Apfel and Seghal et al. in producing nine different combinations. After applying these combinations of mixture laws to 10 tissue phantoms, it is shown that three of the combinations consistently give better predictions of the composition of the phantoms than others. The results verify directly that the phase shift parameter law is probably not a good choice in predicting tissue composition. The potential uses of the mixture methodology are discussed.


Lysosomal instability and cathepsin B release during acetaminophen hepatotoxicity.
Acetaminophen (APAP) overdose is currently the most frequent cause of drug-induced liver failure in the United States. Recently, it was shown that lysosomal iron translocates to mitochondria where it contributes to the collapse of the mitochondrial membrane potential. Therefore, the purpose of this study was to investigate whether cathepsin B, a lysosomal protease, is involved in APAP-induced hepatotoxicity. Cathepsin B activity was measured in subcellular liver fractions of C57Bl/6 mice 3 hr after 300 mg/kg APAP treatment. There was a significant increase in cytoplasmic cathepsin activity, concurrent with a decrease in microsomal activity, indicative of lysosomal cathepsin B release. To investigate the effect of cathepsin B on hepatotoxicity, the cathepsin inhibitor AC-LVK-CHO was given 1 hr prior to 300 mg/kg APAP treatment along with vehicle control. There was no difference between groups in serum alanine aminotransferase (ALT) values, or by histological evaluation of necrosis, although cathepsin B activity was inhibited by 70-80% compared with controls. These findings were confirmed with a different inhibitor (z-FA-fmk) in vivo and in vitro. Hepatocytes were exposed to 5 mM acetaminophen. Lysotracker staining confirmed lysosomal instability and cathepsin B release, but there was no reduction in cell death after treatment with cathepsin B inhibitors. Finally, cathepsin B release was measured in clinical samples from patients with APAP-induced liver injury. Low levels of cathepsin B were released into plasma from overdose patients. APAP overdose causes lysosomal instability and release of cathepsin B into the cytosol but does not contribute to liver injury under these conditions.


Observation of transparency of Erbium-doped silicon nitride in photonic crystal nanobeam cavities.
One dimensional nanobeam photonic crystal cavities are fabricated in an Er-doped amorphous silicon nitride layer. Photoluminescence from the cavities around 1.54 microm is studied at cryogenic and room temperatures at different optical pump powers. The resonators demonstrate Purcell enhanced absorption and emission rates, also confirmed by time resolved measurements. Resonances exhibit linewidth narrowing with pump power, signifying absorption bleaching and the onset of stimulated emission in the material at both 5.5 K and room temperature. We estimate from the cavity linewidths that Er has been pumped to transparency at the cavity resonance wavelength.


Transmission of light through periodic arrays of sub-wavelength slits in metallic hosts.
Using Bloch modes to study the extraordinary transmission of light through a periodic array of slits in a metallic host, we discuss the differing roles of surface plasmon polaritons and Wood's anomalies in the observed behavior of such structures. Under certain circumstances, the first few excited modes appear to play a decisive role in determining the transmission efficiency of the array. Surface plasmon excitations tend to reduce the transmissivity of a semi-infinitely thick slit array, yet, paradoxically, the same reduction can account for enhanced transmission in an array of finite thickness tau, provided that tau is tuned to a Fabry-Perot-like resonance between the entrance and exit facets of the slit array. At the Wood anomaly, power redistribution produces sharp peaks in the diffraction efficiencies of various reflected and transmitted orders of the semi-infinite structure. With skew incidence, the degenerate states split, resulting in two peaks and two valleys, as observed by Wood in his 1902 experiments.


The Nuclear Receptor AhR Controls Bone Homeostasis by Regulating Osteoclast Differentiation via the RANK/c-Fos Signaling Axis.
The aryl hydrocarbon receptor (AhR) pathway plays a key role in receptor activator of NF-κB ligand (RANKL)-mediated osteoclastogenesis. However, the mechanism underlying the regulation of AhR expression in osteoclasts and the signaling pathway through which AhR controls osteoclastogenesis remain unclear. We found that the expression of AhR in bone marrow-derived osteoclasts was upregulated by RANKL at an earlier stage than was the expression of signature osteoclast genes such as those encoding cathepsin K and NFAT, cytoplasmic, calcineurin-dependent 1. In response to RANKL, bone marrow macrophages isolated from AhR-/- mice exhibited impaired phosphorylation of Akt and MAPK as well as NF-κB, whereas their response to M-CSF remained unchanged. Osteoclast differentiation mediated by the AhR signaling pathway was also regulated in an RANKL/c-Fos-dependent manner. Furthermore, ligand activation of AhR by the smoke toxin benzo[a]pyrene accelerated osteoclast differentiation in a receptor-dependent manner, and AhR-dependent regulation of mitochondrial biogenesis in osteoclasts was observed. Moreover, AhR-/- mice exhibited impaired bone healing with delayed endochondral ossification. Taken together, the present results suggest that the RANKL/AhR/c-Fos signaling axis plays a critical role in osteoclastogenesis, thereby identifying the potential of AhR in treating pathological, inflammatory, or metabolic disorders of the bone.


Protein Knotting by Active Threading of Nascent Polypeptide Chain Exiting from the Ribosome Exit Channel.
The mechanism of folding of deeply knotted proteins into their native structure is still not understood. Current thinking about protein folding is dominated by the Anfinsen dogma, stating that the structure of the folded proteins is uniquely dictated by the amino acid sequence of a given protein and that the folding is driven uniquely by the energy gained from interactions between amino acids that contact each other in the native structure of the protein. The role of ribosomes in protein folding was only seen as permitting the folding to progress from the N-terminal part of nascent protein chains. We propose here that ribosomes can participate actively in the folding of knotted proteins by actively threading nascent chains emerging from the ribosome exit channels through loops formed by a synthesized earlier portion of the same protein. Our simulations of folding of deeply knotted protein Tp0624 positively verify the proposed ribosome-driven active threading mechanism leading to the formation of deeply knotted proteins.


Using aquatic animals as partners to increase yield and maintain soil nitrogen in the paddy ecosystems.
Whether species coculture can overcome the shortcomings of crop monoculture requires additional study. Here, we show how aquatic animals (i.e. carp, crabs, and softshell turtles) benefit paddy ecosystems when cocultured with rice. Three separate field experiments and three separate mesocosm experiments were conducted. Each experiment included a rice monoculture (RM) treatment and a rice-aquatic animal (RA) coculture treatment; RA included feed addition for aquatic animals. In the field experiments, rice yield was higher with RA than with RM, and RA also produced aquatic animal yields that averaged 0.52-2.57 t ha-1. Compared to their corresponding RMs, the three RAs had significantly higher apparent nitrogen (N)-use efficiency and lower weed infestation, while soil N contents were stable over time. Dietary reconstruction analysis based on 13C and 15N showed that 16.0-50.2% of aquatic animal foods were from naturally occurring organisms in the rice fields. Stable-isotope-labeling (13C) in the field experiments indicated that the organic matter decomposition rate was greater with RA than with RM. Isotope 15N labeling in the mesocosm experiments indicated that rice used 13.0-35.1% of the aquatic animal feed-N. All these results suggest that rice-aquatic animal coculture increases food production, increases N-use efficiency, and maintains soil N content by reducing weeds and promoting decomposition and complementary N use. Our study supports the view that adding species to monocultures may enhance agroecosystem functions.


A Collimator Setting Optimization Algorithm for Dual-Arc Volumetric Modulated Arc Therapy in Pancreas Stereotactic Body Radiation Therapy.
To optimize collimator setting to improve dosimetric quality of pancreas volumetric modulated arc therapy plan for stereotactic body radiation therapy. Fifty-five volumetric modulated arc therapy cases in stereotactic body radiation therapy of pancreas were retrospectively included in this study with internal review board approval. Different from the routine practice of initializing collimator settings with a template, the proposed algorithm simultaneously optimizes the collimator angles and jaw positions that are customized to the patient geometry. Specifically, this algorithm includes 2 key steps: (1) an iterative optimization algorithm via simulated annealing that generates a set of potential collimator settings from 39 cases with pancreas stereotactic body radiation therapy, and (2) a multi-leaf collimator modulation scoring system that makes the final decision of the optimal collimator settings (collimator angles and jaw positions) based on organs at risk sparing criteria. For validation, the other 16 cases with pancreas stereotactic body radiation therapy were analyzed. Two plans were generated for each validation case, with one plan optimized using the proposed algorithm (Planopt) and the other plan with the template setting (Planconv). Each plan was optimized with 2 full arcs and the same set of constraints for the same case. Dosimetric results were analyzed and compared, including target dose coverage, conformity, organs at risk maximum dose, and modulation complexity score. All results were tested by Wilcoxon signed rank tests, and the statistical significance level was set to .05. Both plan groups had comparable target dose coverage and mean doses of all organs at risk. However, organs at risk (stomach, duodenum, large/small bowel) maximum dose sparing (D0.1 cc and D0.03 cc) was improved in Planopt compared to Planconv. Planopt also showed lower modulation complexity score, which suggests better capability of handling complex shape and sparing organs at risk . The proposed collimator settings optimization algorithm successfully improved dosimetric performance for dual-arc pancreas volumetric modulated arc therapy plans in stereotactic body radiation therapy of pancreas. This algorithm has the capability of immediate clinical application.


The Genomic Health of Ancient Hominins.
The genomes of ancient humans, Neandertals, and Denisovans contain many alleles that influence disease risks. Using genotypes at 3,180 disease-associated loci, we estimated the disease burden of 147 ancient genomes. After correcting for missing data, genetic risk scores (GRS) were generated for nine disease categories and the set of all combined diseases. We used these genetic risk scores to examine the effects of different types of subsistence, geography, and sample age on the number of risk alleles in each ancient genome. On a broad scale, hereditary disease risks are similar for ancient hominins and modern-day humans, and the GRS percentiles of ancient individuals span the full range of what is observed in present-day individuals. In addition, there is evidence that ancient pastoralists may have had healthier genomes than hunter-gatherers and agriculturalists. We also observed a temporal trend whereby genomes from the recent past are more likely to be healthier than genomes from the deep past. This calls into question the idea that modern lifestyles have caused genetic load to increase over time. Focusing on individual genomes, we found that the overall genomic health of the Altai Neandertal is worse than 97% of present-day humans and that Ötzi, the Tyrolean Iceman, had a genetic predisposition for gastrointestinal and cardiovascular diseases. As demonstrated by this work, ancient genomes afford us new opportunities to diagnose past human health, which has previously been limited by the quality and completeness of remains.


Hereditary angioedema: long-term treatment with one or more injections of C1 inhibitor concentrate per week.
Hereditary angioedema due to C1 inhibitor deficiency (HAE-C1-INH) is characterized by recurrent edema attacks in various organs. The objective of the present study was to assess the efficacy and safety of weekly long-term replacement treatment with one or more injections of plasma-derived C1-INH concentrate per week (WLTC) in patients with HAE-C1-INH. Nineteen patients with HAE-C1-INH underwent WLTC for 9 years on average. The benefits and risks were determined based on regular recording by the patients of the severity and number of attacks at the beginning and the end of the study. All patients reported that all or most of their attacks were much less severe: the percentage of severe attacks was 93.3% without and 3.8% with treatment. In 8 of the 14 patients undergoing WLTC, the monthly number of attacks was lower at the end of the study than before the study, higher in 5 patients, and unchanged in 1 patient. The mean number of attacks per week in 6 patients (1 patient initially received weekly on-demand treatment for 2 years and then shifted to WLTC) with weekly on-demand treatment was 4.3 (SD 1.9) at the beginning and 8.0 (SD 3.1) at the end of the study. HAE-C1-INH can be significantly improved by one or more injections of C1-INH concentrate per week. However, patients have to accept a large number of intravenous injections and, in some cases, an increase in disease activity.


Safety incidents in the primary care office setting.
In the United Kingdom, 26% of child deaths have identifiable failures in care. Although children account for 40% of family physicians' workload, little is known about the safety of care in the community setting. Using data from a national patient safety incident reporting system, this study aimed to characterize the pediatric safety incidents occurring in family practice. We undertook a retrospective, cross-sectional, mixed methods study of pediatric reports submitted to the UK National Reporting and Learning System from family practice. Analysis involved detailed data coding using multiaxial frameworks, descriptive statistical analysis, and thematic analysis of a special-case sample of reports. Using frequency distributions and cross-tabulations, the relationships between incident types and contributory factors were explored. Of 1788 reports identified, 763 (42.7%) described harm to children. Three crosscutting priority areas were identified: medication management, assessment and referral, and treatment. The 4 incident types associated with the most harmful outcomes are errors associated with diagnosis and assessment, delivery of treatment and procedures, referrals, and medication provision. Poor referral and treatment decisions in severely unwell or vulnerable children, along with delayed diagnosis and insufficient assessment of such children, featured prominently in incidents resulting in severe harm or death. This is the first analysis of nationally collected, family practice-related pediatric safety incident reports. Recommendations to mitigate harm in these priority areas include mandatory pediatric training for all family physicians; use of electronic tools to support diagnosis, management, and referral decision-making; and use of technological adjuncts such as barcode scanning to reduce medication errors.


Identification of conserved aromatic residues essential for agonist binding and second messenger production at 5-hydroxytryptamine2A receptors.
Several models of agonist binding to G protein-coupled 5-hydroxytryptamine [5-HT] (serotonin) receptors have highlighted the potential importance of highly conserved aromatic residues for ligand binding and agonist efficacy. In this study, we tested these models by constructing and characterizing a number of point mutations of conserved and nonconserved aromatic residues using the 5-HT2A receptor as a model system. Mutations of three highly conserved tryptophans (W200A, W336A, and W367A) proposed to reside near the binding pocket markedly reduced agonist affinity and efficacy at 5-HT2A receptors. Mutations of two other highly conserved aromatic residues postulated to be near the agonist binding site (F340L and Y370A) also had dramatic effects on agonist binding and efficacy. Point mutations of neighboring conserved phenylalanines (F339L and F365L) had minimal effects on agonist binding, although the F365L mutation diminished agonist efficacy. Finally, mutations of two nonconserved aromatic residues (F125L and F383A) not predicted to be near the binding pocket had no effects on agonist binding, potency, or efficacy. Our results are best explained by models that suggest that helices III, V, VI, and VII can form a unit of interacting helices in which highly conserved aromatic residues are oriented toward the center of the helical aggregate to form an aromatic pocket. In addition, our novel results identify a series of aromatic residues essential for agonist-induced second messenger production. These results demonstrate that highly conserved aromatic residues residing in neighboring helices provide the optimum environment for both agonist binding and activation of 5-HT2A receptors.


CHD8 suppresses p53-mediated apoptosis through histone H1 recruitment during early embryogenesis.
The chromodomain helicase DNA-binding (CHD) family of enzymes is thought to regulate gene expression, but their role in the regulation of specific genes has been unclear. Here we show that CHD8 is expressed at a high level during early embryogenesis and prevents apoptosis mediated by the tumour suppressor protein p53. CHD8 was found to bind to p53 and to suppress its transactivation activity. CHD8 promoted the association of p53 and histone H1, forming a trimeric complex on chromatin that was required for inhibition of p53-dependent transactivation and apoptosis. Depletion of CHD8 or histone H1 resulted in p53 activation and apoptosis. Furthermore, Chd8(-/-) mice died early during embryogenesis, manifesting widespread apoptosis, whereas deletion of p53 ameliorated this developmental arrest. These observations reveal a mode of p53 regulation mediated by CHD8, which may set a threshold for induction of apoptosis during early embryogenesis by counteracting p53 function through recruitment of histone H1.


Modulation by acrolein and chloroacetaldehyde of multidrug resistance mediated by the multidrug resistance-associated protein (MRP).
Acrolein (AC) and chloroacetaldehyde (CHA) are metabolites of the non-multidrug resistance cytotoxic drugs cyclophosphamide and ifosfamide. It has previously been reported that both metabolites can induce extensive depletion of glutathione (GSH) in vitro and in vivo and that this depletion occurs at drug concentrations in the micromolar range. A link between the function of the multidrug resistance-associated protein (MRP) and the intracellular concentration of GSH has also been demonstrated. To determine whether AC and CHA can modulate the function of MRP by inducing GSH depletion, we used two human lung cancer cell lines overexpressing MRP: the large cell carcinoma cell line COR-L23/R and the adenocarcinoma cell line MOR/R0.4, along with their respective sensitive parental lines, COR-L23/P and MOR/P. We showed that micromolar concentrations of AC and millimolar concentrations of CHA are able to deplete GSH concentrations in the cell lines studied. In addition, concentrations of 50 micrometer AC and 5 mm CHA could completely reverse the daunorubicin (DNR) and vinblastine accumulation deficit present in COR-L23/R and partially reverse the DNR accumulation deficit in MOR/R0.4. In contrast, AC and CHA did not reverse the drug accumulation deficit in the P-glycoprotein-overexpressing lung cancer cell line H69/LX4. The effect of CHA and AC on drug accumulation was related to the GSH depletion, as we found a concentration-dependent relationship between the GSH levels and the reversal of the accumulation deficit for both AC and CHA. To substantiate further this correlation, we increased cellular GSH content in AC- and CHA-treated cells with the GSH ethyl ester. An increase in cellular GSH levels in CHA- and AC-treated COR-L23/R cells was accompanied by a restoration of the DNR accumulation deficit. No significant effect of the GSH ethyl ester was detected on DNR accumulation in COR-L23/P parental cells. In conclusion, treatment with AC or CHA can reverse the drug accumulation deficit of MRP-overexpressing cells, and this effect appears to be mediated by GSH depletion.