Brazilian-Portuguese version and applicability questionnaire of the mobility index for ankylosing spondyliti.
To translate and adapt the Bath Ankylosing Spondylitis Methodology Index (BASMI) - a metrological measurement for the assessment of patients with ankylosing spondylitis to Brazilian-Portuguese and to analyze the applicability of the questionnaire. The Brazilian-Portuguese version of the Bath Ankylosing Spondylitis Methodology Index was achieved through a translation and back-translation process. This new Bath Ankylosing Spondylitis Methodology Index version was administered to 25 consecutive patients with ankylosing spondylitis who met the 1984 New York criteria for ankylosing spondylitis and were followed in the Soronegative Spondyloarthropathy Unit of the Rheumatology Division, São Paulo University Medical School, from May to November 2005. In order to evaluate the applicability of the questionnaire, it was administered to patients by 2 separate observers, A and B (Rheumatologists), enabling inter-observer test analysis. After 2 weeks, the second interview was administered to the same patients by observer A only to analyze the intra-observer test. All interviews were conducted in the morning. The Pearson coefficient correlation was used to evaluate applicability. There was no conflict stemming from translation and re-translation of Bath Ankylosing Spondylitis Methodology Index questionnaire, and cross-cultural adaptation proved unnecessary. All components had statistically significant coefficients for intra- and inter-observational applicability, with scores ranging from 0.85 to 1.00 and 0.80 to 0.94, respectively. The Brazilian-Portuguese version of the Bath Ankylosing Spondylitis Methodology Index proved to be an applicable instrument for analyzing the mobility index of Brazilian patients with ankylosing spondylitis.


Influence of mirtazapine on salivary cortisol in depressed patients.
Unlike other antidepressants, mirtazapine does not inhibit the reuptake of norepinephrine or serotonin but acts as an antagonist at presynaptic alpha(2)-receptors, at postsynaptic 5-HT2 and 5-HT3 receptors, and at histaminergic H1 receptors. Furthermore, mirtazapine has been shown to acutely inhibit cortisol secretion in healthy subjects. In the present study, the impact of mirtazapine treatment on salivary cortisol secretion was investigated in 12 patients (4 men, 8 women) suffering from major depression according to DSM-IV criteria. Patients were treated with mirtazapine for 3 weeks, receiving 15 mg mirtazapine on day 0, 30 mg on day 1 and 45 mg per day from day 2 up to the end of the study (day 21). Response to mirtazapine treatment was defined by a reduction of at least 50% in the Hamilton Rating Scale for Depression after 3 weeks of therapy. Salivary cortisol concentrations were measured before treatment (day -1), at the beginning of treatment (day 0), after 1 week (day 7) and after 3 weeks (day 21) of treatment with mirtazapine. Saliva samples were collected hourly from 08.00 until 20.00 h. The area under the curve values served as parameter for the salivary cortisol secretion. Following analysis of variance with a repeated measures design, tests with contrasts revealed a significant reduction of cortisol concentrations already after 1 day of mirtazapine treatment that was comparable in responders and nonresponders. In addition to new pharmacological approaches such as CRH1 receptor antagonists, mirtazapine therefore appears to be an effective strategy to decrease hypercortisolism and restore HPA system dysregulation in depression. However, the importance of the acute inhibitory effects of mirtazapine on cortisol secretion for its antidepressant efficacy has to be further clarified.


Hagedorn Temperature of AdS_{5}/CFT_{4} via Integrability.
We establish a framework for calculating the Hagedorn temperature of AdS_{5}/CFT_{4} via integrability. Concretely, we derive the thermodynamic Bethe ansatz equations that yield the Hagedorn temperature of planar N=4 super Yang-Mills theory at any value of the 't Hooft coupling. We solve these equations perturbatively at weak coupling via the associated Y system, confirming the known results at tree level and one-loop order as well as deriving the previously unknown two-loop Hagedorn temperature. Finally, we comment on solving the equations at finite coupling.


Timing and climate forcing of volcanic eruptions for the past 2,500 years.
Volcanic eruptions contribute to climate variability, but quantifying these contributions has been limited by inconsistencies in the timing of atmospheric volcanic aerosol loading determined from ice cores and subsequent cooling from climate proxies such as tree rings. Here we resolve these inconsistencies and show that large eruptions in the tropics and high latitudes were primary drivers of interannual-to-decadal temperature variability in the Northern Hemisphere during the past 2,500 years. Our results are based on new records of atmospheric aerosol loading developed from high-resolution, multi-parameter measurements from an array of Greenland and Antarctic ice cores as well as distinctive age markers to constrain chronologies. Overall, cooling was proportional to the magnitude of volcanic forcing and persisted for up to ten years after some of the largest eruptive episodes. Our revised timescale more firmly implicates volcanic eruptions as catalysts in the major sixth-century pandemics, famines, and socioeconomic disruptions in Eurasia and Mesoamerica while allowing multi-millennium quantification of climate response to volcanic forcing.


[Quantitative mass spectrometry and its utilization in oncology].
Cancers are genetically and clinically very heterogeneous diseases; therefore, various proteomic studies have been trying to find bio-markers which can facilitate prognosis, diagnosis or treatment of these oncological diseases. The mass spectrometry is an effective tool for identification, quantitation, and characterization of biomolecules in the complex bio-logical samples. The first step suitable for selection of bio-markers called discovery proteomics provides a detailed analysis of the samples contributing to the identification of proteins, comparison of their presence in the samples, and selection of the convenient candidates for the prospective bio-markers. The next step of proteomics analysis is directed towards verification of chosen bio-markers with the approach called targeted proteomics. This technique evaluates presence and quantity of the proteins (biomarkers) in clinically precisely defined samples. This article focuses on the description of various approaches suitable for the quantitative analysis of the proteins connected with mass spectrometry.


Evaluation of management of the emergency right hemicolectomy.
To evaluate the morbidity and mortality attending emergency resection of the right colon, we studied retrospectively 119 consecutive patients from two major centers, particularly examining the role of delayed anastomosis (initial ileostomy with subsequent ileocolic anastomosis). Emergency right colectomy was defined as an operative procedure in which partial resection of the terminal ileum and total or partial resection of the cecum and/or ascending colon were necessary before the colon could be evacuated and prepared as might be the case in elective resection. Ninety patients underwent primary anastomosis; hospital mortality and morbidity were 26 and 11%, respectively. Among 29 patients who underwent ileostomy, hospital mortality was 38%; hospital morbidity was 10%. In general, the patients selected for ileostomy were somewhat more ill or had more severe injury. only 17% of all deaths and 8% of all serious complications could be attributed to the anastomosis, leaving little statistical opportunity for improvement by deleting it. The data indicate that a very high mortality (29%) attends emergency right hemicolectomy. Our study indicates that ileostomy in lieu of anastomosis does not reduce this high mortality.


The ubiquitin ligase gp78 promotes sarcoma metastasis by targeting KAI1 for degradation.
Metastasis is the primary cause of mortality from cancer, but the mechanisms leading to metastasis are poorly understood. In particular, relatively little is known about metastasis in cancers of mesenchymal origins, which are known as sarcomas. Approximately ten proteins have been characterized as 'metastasis suppressors', but how these proteins function and are regulated is, in general, not well understood. Gp78 (also known as AMFR or RNF45) is a RING finger E3 ubiquitin ligase that is integral to the endoplasmic reticulum (ER) and involved in ER-associated degradation (ERAD) of diverse substrates. Here we report that expression of gp78 has a causal role in the metastasis of an aggressive human sarcoma and that this prometastatic activity requires the E3 activity of gp78. Further, gp78 associates with and targets the transmembrane metastasis suppressor, KAI1 (also known as CD82), for degradation. Suppression of gp78 increases KAI1 abundance and reduces the metastatic potential of tumor cells, an effect that is largely blocked by concomitant suppression of KAI1. An inverse relationship between these proteins was confirmed in a human sarcoma tissue microarray. Whereas most previous efforts have focused on genetic mechanisms for the loss of metastasis suppressor genes, our results provide new evidence for post-translational downregulation of a metastasis suppressor by its ubiquitin ligase, resulting in abrogation of its metastasis-suppressing effects.


Hormonal influences on mammary tumors of the rat. I. Acceleration of growth of transplanted fibroadenoma in ovariectomized and hypophysectomized rats.
A transplanted mammary fibroadenoma was found to grow in 95 per cent of intact adult female rats and the increment of tumor weights was progressive and logarithmic. The growth of the tumor was retarded by ovariectomy and still more when this was combined with adrenalectomy. In ovariectomized rats the growth of the tumor was stimulated by phenolic estrogens, this increase being enhanced when progesterone was added. In these responses to hormonal changes the mammary gland and the tumor resembled each other. Yet there are many differences between the growth of the fibroadenoma and that of the mammary gland. In contrast to the progressive growth which occurred in intact adult females there was a prolonged period of indolent growth of transplants in hypophysectomized rats; but after many weeks active growth began and the tumors eventually reached large size. During the period of quiescent growth the tumor was cytologically atrophic but after the growth spurt had started the microscopic appearance of the fibroadenoma resembled that of tumors growing in normal adult females. The mammary gland remained atrophic during both the slow and the accelerated phases of tumor growth, and so too with the other secondary sex expressions. In hypophysectomized rats estrone and progesterone, when combined, stimulated the growth of the tumor, and this growth was accelerated by the additional administration of lactogenic or growth hormones. None of these hormones, separately, stimulated the growth of the tumor. In ovariectomized rats other differences were demonstrated between the growth of the mammary gland and the fibroadenoma. Progesterone, injected alone, accelerated the growth of the tumor but not that of the mammary glands. The administration of phenolic estrogens exerted a biphasic effect on the growth of the tumor whilst that on the breast of its hosts was monophasic. With progressively increasing doses of these phenols there occurred primarily an augmentation of the rate of growth of the tumor until a peak was achieved; an increase of the dose above the optimal amount depressed the growth of the tumor. The stage of depression of growth was not observed in the mammary glands of these tumor-bearing rats. Many steroids which induced gestational changes in the mammary gland accelerated the growth of the tumor. Among these were estrone and progesterone in combination and 17alpha-ethinyl-19-nor-testosterone administered alone. But gestational changes developed in the mammary gland of rats treated with 4-androstene-3alpha,17beta-diol, without growth of the tumor. The evidence which we have presented proves that the mammary fibroadenoma tested had some of the functional properties of a normal mammary gland, and neoplastic traits as well. In its response to hormones it had characteristics which set it apart from all other endocrine targets of the rat.


A sporulation-specific, sigF-dependent protein, SspA, affects septum positioning in Streptomyces coelicolor.
The RNA polymerase sigma factor SigF controls late development during sporulation in the filamentous bacterium Streptomyces coelicolor. The only known SigF-dependent gene identified so far, SCO5321, is found in the biosynthetic cluster encoding spore pigment synthesis. Here we identify the first direct target for SigF, the gene sspA, encoding a sporulation-specific protein. Bioinformatic analysis suggests that SspA is a secreted lipoprotein with two PepSY signature domains. The sspA deletion mutant exhibits irregular sporulation septation and altered spore shape, suggesting that SspA plays a role in septum formation and spore maturation. The fluorescent translational fusion protein SspA-mCherry localized first to septum sites, then subsequently around the surface of the spores. Both SspA protein and sspA transcription are absent from the sigF null mutant. Moreover, in vitro transcription assay confirmed that RNA polymerase holoenzyme containing SigF is sufficient for initiation of transcription from a single sspA promoter. In addition, in vivo and in vitro experiments showed that sspA is a direct target of BldD, which functions to repress sporulation genes, including whiG, ftsZ and ssgB, during vegetative growth, co-ordinating their expression during sporulation septation.


The long shadow of work - does time since labour market exit affect the association between socioeconomic position and health in a post-working population.
To test the effect of time since labour market exit (LME) on associations between socioeconomic position (SEP) and self-rated health. Retirees from the English Longitudinal Study of Ageing (ELSA) were divided into three groups on the basis of the length of time since LME. Seven different indicators of SEP were identified: socioeconomic class, income, wealth, education, tenure, area deprivation and subjective social status. Unadjusted and mutually adjusted logistic regression analyses were performed with poor self-rated health as the outcome. The sample consisted of 2617 men (mean (SD) age 71.69 (7.04) years) and 2619 women (71.29 (8.26) years). In the unadjusted analyses, patterns of association between SEP measures and health were similar for men and women. Most SEP measures were associated with poor health, although the effects were attenuated by time since LME. In the mutually adjusted analyses, wealth was found to have a strong independent effect on health among men, especially in those groups that left the labour market < or = 20 years ago,while for women subjective social status seemed to have the most important effect on health after LME. Time since LME is an important factor to consider when studying health inequalities in a post-working population. The effect of time since LME varies according to gender and the measures of SEP used. Further work in this area should take account of age, period and cohort effects using multiple measures of SEP and more refined measures of LME.


Predictors of prolonged length of intensive care unit stay after stage I palliation: a report from the National Pediatric Cardiology Quality Improvement Collaborative.
The objective of this study is to identify predictors of prolonged intensive care unit (ICU) length of stay (LOS) for single ventricle patients following Stage I palliation. We hypothesize that peri-operative factors contribute to prolonged ICU stay among children with hypoplastic left heart syndrome (HLHS) and its variants. In 2008, as a part of the Joint Council on Congenital Heart Disease initiative, the National Pediatric Cardiology-Quality Improvement Collaborative established a data registry for patients with HLHS and its variants undergoing staged palliation. Between July 2008 and August 2011, 33 sites across the United States submitted discharge data essential to this analysis. Data describing the patients, their procedures, and their hospital experience were entered. LOS estimates were generated. Prolonged LOS in the ICU was defined as stay greater than or equal to 26 days (i.e., 75th percentile). Statistical analyses were carried out to identify pre-operative, operative, and post-operative predictors of prolonged LOS in the ICU. The number of patients with complete discharge data was 303, and these subjects were included in the analysis. Univariate and multivariate analyses were performed. Multivariate analysis revealed that lower number of enrolled participants (e.g., 1-10) per site, the presence of pre-operative acidosis, increased circulatory arrest time, the occurrence of a central line infection, and the development of respiratory insufficiency requiring re-intubation were associated with prolonged LOS in the ICU. Prolonged LOS in the ICU following Stage I palliation in patients with HLHS and HLHS variant anatomy is associated with site enrollment, circulatory arrest time, pre-operative acidosis, and some post-operative complications, including central line infection and re-intubation. Further study of these associations may reveal strategies for reducing LOS in the ICU following the Norwood and Norwood-variant surgeries.


Use of Population-Level Administrative Data in Developmental Science.
Population-level administrative data-data on individuals' interactions with administrative systems (e.g., health, criminal justice, and education)-have substantially advanced our understanding of life-course development. In this review, we focus on five areas where research using these data has made significant contributions to developmental science: (a) understanding small or difficult-to-study populations, (b) evaluating intergenerational and family influences, (c) enabling estimation of causal effects through natural experiments and regional comparisons, (d) identifying individuals at risk for negative developmental outcomes, and (e) assessing neighborhood and environmental influences. Further advances will be made by linking prospective surveys to administrative data to expand the range of developmental questions that can be tested; supporting efforts to establish new linked administrative data resources, including in developing countries; and conducting cross-national comparisons to test findings' generalizability. New administrative data initiatives should involve consultation with population subgroups including vulnerable groups, efforts to obtain social license, and strong ethical oversight and governance arrangements.


Preoperative nomogram to predict the likelihood of complications after radical nephroureterectomy.
To construct a nomogram based on preoperative variables to better predict the likelihood of complications occurring within 30 days of radical nephroureterectomy (RNU). The charts of 731 patients undergoing RNU at eight academic medical centres between 2002 and 2014 were reviewed. Preoperative clinical, demographic and comorbidity indices were collected. Complications occurring within 30 days of surgery were graded using the modified Clavien-Dindo scale. Multivariate logistic regression determined the association between preoperative variables and post-RNU complications. A nomogram was created from the reduced multivariate model with internal validation using the bootstrapping technique with 200 repetitions. A total of 408 men and 323 women with a median age of 70 years and a body mass index of 27 kg/m2 were included. A total of 75% of the cohort was white, 18% had an Eastern Cooperative Oncology Group (ECOG) performance status ≥2, 20% had a Charlson comorbidity index (CCI) score >5 and 50% had baseline chronic kidney disease (CKD) ≥ stage III. Overall, 279 patients (38%) experienced a complication, including 61 events (22%) with Clavien grade ≥ III. A multivariate model identified five variables associated with complications, including patient age, race, ECOG performance status, CKD stage and CCI score. A preoperative nomogram incorporating these risk factors was constructed with an area under curve of 72.2%. Using standard preoperative variables from this multi-institutional RNU experience, we constructed and validated a nomogram for predicting peri-operative complications after RNU. Such information may permit more accurate risk stratification on an individual cases basis before major surgery.


Cancer Advisory Board.
The article "Reagan seeks expansion of Soviet ties" by R. Jeffrey Smith (News and Comment, 13 July, p. 145) erroneously stated that the U.S. Senate had passed a resolution calling for a U.S.-Soviet summit "without preconditions or assurances of success." Shortly before midnight on 19 June, the resolution was amended to call for a summit at "the earliest practical time following thorough preparation." It was approved in this form.


Conservative treatment of favorable rectal carcinomas.
Conservative treatment of rectal cancer with either full thickness en bloc local excision or endocavitary irradiation provides excellent local control for highly selected early rectal carcinomas. For either approach the selection criteria are similar. An ideal lesion should be: 1) freely mobile; 2) less than 3 cm in diameter; 3) no evidence of extension into the perirectal fat either on digital examination or on transrectal US; 4) no clinical evidence of involved nodes on physical examination of diagnostic studies; 5) the surgical alternative would be an abdominal perineal resection; 6) well or moderately differentiated. External beam radiation therapy can contribute to the results of conservative treatment by improving the chances of controlling subclinical nodal disease in the pelvis as well as by improving the control of disease in the rectum at the margins of the local procedure. For highly selected patients, the local control and disease-free interval is on the order of 90% with either endocavitary radiation or local excision.


A proteomic approach to identifying proteins differentially expressed in conidia and mycelium of the entomopathogenic fungus Metarhizium acridum.
Metarhizium spp. is an important worldwide group of entomopathogenic fungi used as an interesting alternative to chemical insecticides in programs of agricultural pest and disease vector control. Metarhizium conidia are important in fungal propagation and also are responsible for host infection. Despite their importance, several aspects of conidial biology, including their proteome, are still unknown. We have established conidial and mycelial proteome reference maps for Metarhizium acridum using two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF MS). In all, 1130±102 and 1200±97 protein spots were detected in ungerminated conidia and fast-growing mycelia, respectively. Comparison of the two protein-expression profiles reveled that only 35% of the protein spots were common to both developmental stages. Out of 94 2-DE protein spots (65 from conidia, 25 from mycelia and two common to both) analyzed using mass spectrometry, seven proteins from conidia, 15 from mycelia and one common to both stages were identified. The identified protein spots exclusive to conidia contained sequences similar to known fungal stress-protector proteins (such as heat shock proteins (HSP) and 6-phosphogluconate dehydrogenase) plus the fungal allergen Alt a 7, actin and the enzyme cobalamin-independent methionine synthase. The identified protein spots exclusive to mycelia included proteins involved in several cell housekeeping biological processes. Three proteins (HSP 90, 6-phosphogluconate dehydrogenase and allergen Alt a 7) were present in spots in conidial and mycelial gels, but they differed in their locations on the two gels.


Predatory aquatic beetles, suitable trace elements bioindicators.
Predatory aquatic beetles are common colonizers of natural and managed aquatic environments. While as important components of the aquatic food webs they are prone to accumulate trace elements, they have been largely neglected from metal uptake studies. We aim to test the suitability of three dytiscid species, i.e.Hydroglyphus pusillus, Laccophilus minutus and Rhantus suturalis, as trace elements (Al, As, Cd, Co, Cu, Fe, Mn, Mo, Ni, Pb, Se and Zn) bioindicators. The work was carried out in a case area representing rice paddies and control sites (reservoirs) from an arid region known for its land degradation (Monegros, NE Spain). Categorical principal component analysis (CATPCA) was tested as a nonlinear approach to identify significant relationships between metals, species and habitat conditions so as to examine the ability of these species to reflect differences in metal uptake. Except Se and As, the average concentrations of all other elements in the beetles were higher in the rice fields than in the control habitats. The CATPCA determined that H. pusillus had high capacity to accumulate Fe, Ni and Mn regardless of the habitat type, and hence may not be capable of distinguishing habitat conditions with regards to these metals. On the other hand, L. minutus was found less sensitive for Se in non-managed habitats (i.e. reservoirs), while R. suturalis was good in accumulating Al, Mo and Pb in rice fields. The latter seems to be a promising bioindicator of metal enrichment in rice fields. We conclude that predatory aquatic beetles are good candidates for trace elements bioindication in impacted and non-impacted environments and can be used in environmental monitoring studies. CATPCA proved to be a reliable approach to unveil trends in metal accumulation in aquatic invertebrates according to their habitat status.


A sensitive and rapid method to determine the viability of freeze-dried bacterial cells.
To determine the potential use of flow cytometry for viability asssessment of freeze-dried bacterial cells. Escherichia coli CIP 54.8T and Vibrio metschnikovii CIP 104262 were analysed. The viability of freeze-dried cells resuspended in a nutrient broth was evaluated by culture whereas activity was determined by flow cytometry analysis of both esterase activity and cell death. Activity assessment by flow cytometry was found to be a rapid and good indicator of cell viability and was very efficient for quality control. For V. metschnikovii the fraction of active cells varies greatly depending on the freeze-drying procedure and within a given procedure. Bacterial activity assessment by flow cytometry is very efficient for the control of freeze-dried cells.


A comparison of sleep and daytime sleepiness in depressed and non-depressed mothers during the early postpartum period.
Taiwanese mothers have identified insufficient sleep as a major manifestation of postpartum depression. Few studies have thoroughly examined the relationship between sleep and depression during the early postpartum period, however. The objectives of this study were to compare the characteristics of both the postpartum sleep and daytime sleepiness of depressed first-time mothers and of their non-depressed counterparts, and to determine the factors that significantly increased mothers' risks of being depressed. A non-probability sample of 163 first-time mothers completed a questionnaire between the 13th and 20th days of the postpartum period. The Center of Epidemiological Studies- Depression and Pittsburgh Sleep Quality Index were used to measure mothers' experiences of depression symptoms and sleep. Daytime sleepiness was estimated in four ways, derived from the Roy Adaptation Model. The results indicated that the depressed mothers had poorer sleep quality than the non-depressed mothers, slept less efficiently, reported more sleep disturbances, and exhibited more daytime dysfunctions. Mothers who frequently perceived their daytime sleepiness to be affected by infant-care performance were more likely to be depressed. The study' s findings support the view that there is a connection between depression and poor sleep among postpartum mothers in Taiwan, and indicate that depressed mothers' experiences of poor sleep are multi-faceted, and not simply a matter of insufficient sleep.


Effects of partner proteins on BCA2 RING ligase activity.
BCA2 is an E3 ligase linked with hormone responsive breast cancers. We have demonstrated previously that the RING E3 ligase BCA2 has autoubiquitination activity and is a very unstable protein. Previously, only Rab7, tetherin, ubiquitin and UBC9 were known to directly interact with BCA2. Here, additional BCA2 binding proteins were found using yeast two-hybrid and bacterial-II-hybrid screening techniques with Human breast and HeLa cDNA libraries. Co-expression of these proteins was analyzed through IHC of TMAs. Investigation of the molecular interactions and effects were examined through a series of in vivo and in vitro assays. Ten unique BCA2 interacting proteins were identified, two of which were hHR23a and 14-3-3sigma. Both hHR23a and 14-3-3sigma are co-expressed with BCA2 in breast cancer cell lines and patient breast tumors (n = 105). hHR23a and BCA2 expression was significantly correlated (P = < 0.0001 and P = 0.0113) in both nucleus and cytoplasm. BCA2 expression showed a statistically significant correlation with tumor grade. High cytoplasmic hHR23a trended towards negative nodal status. Binding to BCA2 by hHR23a and 14-3-3sigma was confirmed in vitro using tagged partner proteins and BCA2. hHR23a and 14-3-3sigma effect the autoubiquitination and auto-degradation activity of BCA2. Ubiquitination of hHR23a-bound BCA2 was found to be dramatically lower than that of free BCA2, suggesting that hHR23a promotes the stabilization of BCA2 by inactivating its autoubiquitination activity, without degradation of hHR23a. On the other hand, phosphorylated BCA2 protein is stabilized by interaction with 14-3-3sigma both with and without proteasome inhibitor MG-132 suggesting that BCA2 is regulated by multiple degradation pathways. The interaction between BCA2 and hHR23a in breast cancer cells stabilizes BCA2. High expression of BCA2 is correlated with grade in breast cancer, suggesting regulation of this E3 ligase is important to cancer progression.


Structure and functional properties of Bacillus subtilis endospore biogenesis factor StoA.
Bacillus subtilis StoA is an extracytoplasmic thiol-disulfide oxidoreductase (TDOR) important for the synthesis of the endospore peptidoglycan cortex protective layer. Here we demonstrate that StoA is membrane-associated in B. subtilis and report the crystal structure of the soluble protein lacking its membrane anchor. This showed that StoA adopts a thioredoxin-like fold with N-terminal and internal additions that are characteristic of extracytoplasmic TDORs. The CXXC active site of the crystallized protein was found to be in a mixture of oxidized and reduced states, illustrating that there is little conformational variation between redox states. The midpoint reduction potential was determined as -248 mV versus normal hydrogen electrode at pH 7 consistent with StoA fulfilling a reductive role in endospore biogenesis. pK(a) values of the active site cysteines, Cys-65 and Cys-68, were determined to be 5.5 and 7.8. Although Cys-68 is buried within the structure, both cysteines were found to be accessible to cysteine-specific alkylating reagents. In vivo studies of site-directed variants of StoA revealed that the active site cysteines are functionally important, as is Glu-71, which lies close to the active site and is conserved in many reducing extracytoplasmic TDORs. The structure and biophysical properties of StoA are very similar to those of ResA, a B. subtilis extracytoplasmic TDOR involved in cytochrome c maturation, raising important general questions about how these similar but non-redundant proteins achieve specificity. A detailed comparison of the two proteins demonstrates that relatively subtle differences, largely located around the active sites of the proteins, are sufficient to confer specificity.


Inhibition of the cardiac angiogenic response to exogenous vascular endothelial growth factor.
The angiogenic effects of vascular endothelial growth factor (VEGF) are mediated by the stimulation of endothelial nitric oxide synthase (eNOS) and nitric oxide release. Nitric oxide availability is decreased in patients with coronary disease, a possible explanation for the humble results of VEGF in clinical trials. We sought to examine the effects of exogenous VEGF in a model of endothelial dysfunction. Miniswine fed either a regular (N = 6, group NORM) or hypercholesterolemic diet (N = 6, HICHOL) underwent ameroid placement on the circumflex artery. Three weeks later, baseline myocardial perfusion was assessed by microsphere injections, and all pigs were treated with VEGF. Four weeks later, microsphere injections were repeated and the hearts harvested. Endothelial-dependent coronary microvascular reactivity, and VEGF and eNOS expression were assessed. HICHOL pigs showed significant endothelial dysfunction in the ischemic territory. Post-treatment myocardial blood flow in the circumflex territory was significantly higher in the NORM compared to the HICHOL group. VEGF and eNOS levels were increased in the ischemic territory in the NORM group but decreased in the HICHOL group. The cardiac angiogenic response to VEGF was markedly inhibited in a hypercholesterolemia-induced porcine model of endothelial dysfunction. Coronary endothelial dysfunction could be an obstacle to the efficacy of clinical angiogenesis protocols and a putative therapeutic target.


Aging of tissue-resident adult stem/progenitor cells and their pathological consequences.
The fascinating discovery of tissue-resident adult stem/progenitor cells in recent years led to an explosion of interest in the development of novel stem cell-based therapies for improving the regenerative capacity of these endogenous immature cells or transplanted cells for the repair of damaged and diseased tissues. In counterbalance, a growing body of evidence has revealed that the changes in phenotypic and functional properties of human adult stem/progenitor cells may occur during chronological aging and have severe pathological consequences. Especially, intense oxidative and metabolic stress and chronic inflammation, enhanced telomere attrition and defects in DNA repair mechanisms may lead to severe DNA damages and genomic instability in adult stem/progenitor cells with advancing age that may in turn trigger their replicative senescence and/or programmed cell death. Moreover, the changes in the intrinsic and extrinsic factors involved in the stringent control of self-renewal and multilineage differentiation capacities of these regenerative cells, including deregulated signals from the aged niche, may also contribute to their dysfunctions or loss during chronological aging. This age-associated decline in the regenerative capacity and number of functional adult stem/progenitor cells may increase the risk to develop certain diseases. At opposed end, the telomerase reactivation and accumulation of genetic alterations leading to a down-regulation of numerous tumor suppressor genes concomitant with the enhanced expression of diverse oncogenic products may result in their malignant transformation into cancer-initiating cells. Therefore, the rescue or replacement of aged and dysfunctional endogenous adult stem/progenitor cells or molecular targeting of their malignant counterpart, cancer stem/progenitor cells may constitute potential anti-aging and cancer therapies. These therapeutic strategies could be used for treating diverse devastating premature aging and age-related disorders including hematopoietic and immune disorders, heart failure and cardiovascular diseases, neurodegenerative, muscular and gastrointestinal diseases, atherosclerosis and aggressive and lethal cancers.


Wildland firefighter deaths in the United States: A comparison of existing surveillance systems.
Wildland fire fighting is a high-risk occupation requiring considerable physical and psychological demands. Multiple agencies publish fatality summaries for wildland firefighters; however, the reported number and types vary. At least five different surveillance systems capture deaths, each with varying case definitions and case inclusion/exclusion criteria. Four are population-level systems and one is case-based. System differences create challenges to accurately characterize fatalities. Data within each of the five surveillance systems were examined to better understand the types of wildland firefighter data collected, to assess each system's utility in characterizing wildland firefighter fatalities, and to determine each system's potential to inform prevention strategies. To describe similarities and differences in how data were recorded and characterized, wildland fire deaths for three of the population-based systems were matched and individual fatalities across systems were compared. Between 2001 and 2012, 247 unique deaths were captured among the systems; 73% of these were captured in all three systems. Most common causes of death in all systems were associated with aviation, vehicles, medical events, and entrapments/burnovers. The data show that, although the three systems often report similar annual summary statistics, events captured in each system vary each year depending on the types of events that the system is designed to track, such as inclusion/exclusion of fatalities associated with the Hometown Heroes Survivor Benefits Act of 2003. The overarching and central goal of each system is to collect accurate and timely information to improve wildland firefighter safety and health. Each system is unique and has varying inclusion and exclusion criteria for capturing and tracking different subsets of wildland firefighter tasks and duties. Use of a common case definition and better descriptions and interpretations of the data and the results would help to more accurately characterize wildland firefighter traumatic injuries and illnesses, lessen the likelihood for misinterpretation of wildland firefighter fatality data, and assist with defining the true occupational injury burden within this high-risk population.


Differential expression of the human metastasis adhesion molecule CD44V in normal and carcinomatous stomach mucosa of Chinese subjects.
CD44 is a cell surface adhesion molecule involved in cell-cell and cell-matrix interactions. Tumor cells transfected to overexpress the isoform CD44V readily gain access to lymph nodes and form distant metastases. Monoclonal antibodies directed at epitopes common to known CD44 isoforms were used to investigate CD44V expression in 30 normal gastric mucosa tissues, 64 different gastric adenocarcinomas, 20 metastatic adenocarcinoma lymph nodes and 4 established gastric carcinoma cell lines. In addition, CD44V gene expression in six gastric adenocarcinoma tissues and four gastric cancer cell lines were investigated by northern blotting. Immunohistochemistry screening of 30 subjects with normal gastric mucosa did not reveal expression of CD44 variants. Areas of intestinal metaplasia, a precancerous lesion, were stained with antibodies against either V5- or V6-containing isoforms of CD44. Tubular and signet-ring cell types of adenocarcinoma were strongly positive for epitopes encoded by CD44 variants containing exons V5 (41/49 and 10/10, respectively). Some tubular type adenocarcinomas (15/49) also expressed CD44 variants containing the V6 epitope. Tumor differentiation was closely related to CD44 V5 expression (P < 0.001). In addition, 18 of 20 gastric adenocarcinomas metastatic to lymph nodes expressed the V5 epitope of CD44 and 4 of 20 expressed the V6 epitope. Analysis of four established gastric adenocarcinoma cell lines revealed that two had moderate to strong expression of exons V5 and V6 of CD44. An antibody directed against CD44 variants containing exons V8 to V10 strongly stained all gastric adenocarcinoma cell lines. Northern blotting demonstrated that all four tumor cell lines and six gastric carcinoma mucosa tissues expressed CD44V. Generation of CD44 splice variants may be closely linked with gastric carcinoma tumorigenesis and differentiation. In addition, expression of CD44 variants containing exons V5 and V6 may be used as an indicator of evolving gastric cancer.


Slitlamp, specular, and light microscopic findings of human donor corneas after laser-assisted in situ keratomileusis.
To examine slitlamp, specular, and light microscopic features of human donor corneas known to have undergone laser-assisted in situ keratomileusis (LASIK). Twenty-six donor corneas known to have undergone LASIK prospectively underwent slitlamp examination with particular attention to the presence of a flap edge, as well as specular microscopy with particular attention to the presence of highly reflective particles in the stroma corresponding to the LASIK interface. Central endothelial cell density and pachymetery were obtained. They were compared with 26 control donor corneas without LASIK. Eleven LASIK donor corneas were processed for histology. Twenty-six donor corneas with no known prior keratorefractive surgery also underwent similar slitlamp examination and specular microscopy to serve as controls. Twelve (46%) of 26 LASIK donor corneas had an obvious flap edge, and 10 (39%) had a subtle flap edge by slitlamp examination. Four (15%) had infiltrates by slitlamp examination, of which 3 were confirmed by histopathologic examination. Highly reflective particles were seen by specular microscopy in the stroma of 23 (88%) of 26 LASIK donor corneas, but only 1 (4%) of 26 control donor corneas had a single highly reflective particle in the stroma (P<.001). The mean central endothelial cell counts were similar: 2138 cells/mm(2) in the LASIK group compared with 2250 cells/mm(2) in the controls (P =.39). Vacuolization and pyknosis of keratocytes was a consistent histopathologic finding after LASIK. Metallic particles at the interface were not detected by histology. Detection of a flap edge by slitlamp examination may detect at least half of the donor corneas that may have undergone LASIK. The detection of highly reflective stromal particles may form an effective basis for screening for LASIK donor corneas using specular microscopy and requires further study.


Automatic Annotation of Subsea Pipelines using Deep Learning.
Regulatory requirements for sub-sea oil and gas operators mandates the frequent inspection of pipeline assets to ensure that their degradation and damage are maintained at acceptable levels. The inspection process is usually sub-contracted to surveyors who utilize sub-sea Remotely Operated Vehicles (ROVs), launched from a surface vessel and piloted over the pipeline. ROVs capture data from various sensors/instruments which are subsequently reviewed and interpreted by human operators, creating a log of event annotations; a slow, labor-intensive and costly process. The paper presents an automatic image annotation framework that identifies/classifies key events of interest in the video footage viz. exposure, burial, field joints, anodes, and free spans. The reported methodology utilizes transfer learning with a Deep Convolutional Neural Network (ResNet-50), fine-tuned on real-life, representative data from challenging sub-sea environments with low lighting conditions, sand agitation, sea-life and vegetation. The network outputs are configured to perform multi-label image classifications for critical events. The annotation performance varies between 95.1% and 99.7% in terms of accuracy and 90.4% and 99.4% in terms of F1-Score depending on event type. The performance results are on a per-frame basis and corroborate the potential of the algorithm to be the foundation for an intelligent decision support framework that automates the annotation process. The solution can execute annotations in real-time and is significantly more cost-effective than human-only approaches.


The location and mechanism of hyperosmotic fluid secretion in the rectum of the saline-water mosquito larvae Aedes taeniorhynchus.
1. Ligation between the anterior and posterior segments of the rectum in vitro was used to demonstrate that the posterior rectum is the site of hyperosmotic secretion to the lumen side. Observations were consistent with a reabsorptive function for the anterior rectum. These results support predictions from ultrastructural studies of these two segments. 2. The initial potential of the rectal lumen, relative to the haemocoel side, was of opposite polarity in the anterior (-10 mV) and posterior (+ 10 mV) segments and these values decreased to -2 and +6 mV respectively in ligated recta which had secreted for 2 h. 3. A comparison of these potential difference measurements with concentration differences developed across the rectal epithelium under the same experimental conditions indicates that Na+, K+, Mg2+, and Cl- are all actively transported by the posterior segment to the lumen side. 4. The influence of different haemolymph concentrations of Na+, K+, and Cl- on the potential differences across the basal cell border and across the whole rectal epithelium are reported. Based on this and previous data, we propose a model for the organization of transport processes within the single celltype present in the posterior rectal epithelium.


Automated platform for cell selection and separation based on four-dimensional motility and matrix degradation.
Motility and invasion are key steps in the metastatic cascade, enabling cells to move through normal tissue borders into the surrounding stroma. Most available in vitro assays track cell motility or cell invasion but lack the ability to measure both simultaneously and then separate single cells with unique behaviors. In this work, we developed a cell-separation platform capable of tracking cell movement (chemokinesis) and invasion through an extracellular matrix in space and time. The platform utilized a collagen scaffold with embedded tumor cells overlaid onto a microraft array. Confocal microscopy enabled high resolution (0.4 × 0.4 × 3.5 µm voxel) monitoring of cell movement within the scaffolds. Two pancreatic cancer cell lines with known differing invasiveness were characterized on this platform, with median motilities of 14 ± 6 μm and 10 ± 4 μm over 48 h. Within the same cell line, cells demonstrated highly variable motility, with XYZ movement ranging from 144 μm to 2 μm over 24 h. The ten lowest and highest motility cells, with median movements of 33 ± 11 μm and 3 ± 1 μm, respectively, were separated and sub-cultured. After 6 weeks of culture, the cell populations were assayed on a Transwell invasion assay and 227 ± 56 cells were invasive in the high motility population while only 48 ± 10 cells were invasive in the low motility population, indicating that the resulting offspring possessed a motility phenotype reflective of the parental cells. This work demonstrates the feasibility of sorting single cells based on complex phenotypes along with the capability to further probe those cells and explore biological phenomena.


Analgesic activity of the aqueous fraction from the ethanolic extract of Chrysanthemum indicum in mice.
The aqueous fraction (AF) of an ethanolic extract from Chrysanthemum indicum was evaluated for analgesic activity in mice using chemical and thermal models of nociception. Given orally, AF at doses of 300 and 600 mg/kg produced significant inhibitions on chemical nociception induced by intraperitoneal acetic acid, subplantar formalin/capsaicin injections and on thermal nociception in the tail-flick test and in the hot plate test. In the pentobarbital sodium-induced sleeping time test and the open-field test, AF neither significantly enhanced the pentobarbital sodium-induced sleeping time nor impaired the motor performance, indicating that the observed analgesic activity was unlikely due to sedation or motor abnormality. In a measurement of core body temperature, AF did not affect temperature within 80 min. Moreover, the effective dose (600 mg/kg) also showed no toxicity within 7 days. These results suggested further that AF produced analgesic activity possibly related to the flavonoid glycosides and phenolic glycosides in this fraction.


Guidelines for the diagnosis and treatment of coronavirus disease 2019 (COVID-19) in China.
With the deepening of the understanding and research in coronavirus disease 2019 (COVID-19), the diagnosis and treatment of COVID-19 have been constantly updated and improved. In China, since the implementation of "Guidelines for the Diagnosis and Treatment of COVID-19 (1st Trial Version)" on Jan. 15, 2020, 2nd to 7th versions (including revision of 5th version) was updated from Jan. 18, Jan. 22, Jan. 27, Feb. 4, Feb. 8, Feb. 18 and Mar. 3, respectively. Versions updated subsequently provide more detailed information in many ways than the 1st and 2nd versions, so this paper will introduce the development of the main contents of the 3rd to 7th versions of COVID-19 guidelines in China, which hopes to provide help for clinical medical staff in other countries fighting with this disease.


Cardiovascular effects of capsaicin-sensitive neurons.
Calcitonin gene-related peptide, the main transmitter released from capsaicin-sensitive sensory-motor fibers, has positive inotropic and chronotropic effects on the heart and causes vasodilatation in the coronary arteries and elsewhere in the peripheral vasculature. We review some aspects of the cardiovascular actions induced by exogenous calcitonin gene-related peptide and by release of the peptide following activation of capsaicin-sensitive nerves. The efferent function of cardiac sensory-motor neurones is modulated by a number of endogenous substances of physiopathological interest, including opioid peptides, norepinephrine and adenosine. The receptors involved in the prejunctional regulation due to these substances have been characterized. Studies on the mesenteric bed of the rat have shown that, at least in such a resistance vascular bed, the relaxing effect of calcitonin gene-related peptide is mediated by a direct mechanism independent of endothelium-derived nitric oxide release. In cultured human endothelial cells from the umbilical cord vein, calcitonin gene-related peptide, at nM concentrations, stimulates cell growth in a dose-dependent manner. The possible implications of calcitonin gene-related peptide in the physio-pathological regulation of the cardiovascular system and in the trophism of vascular tissues are discussed.


Separation of polypeptides by isoelectric point focusing in electrospray-friendly solution using a multiple-junction capillary fractionator.
We introduce an online multiple-junction capillary isoelectric focusing fractionator (OMJ-CIEF) for separation of biological molecules in solution by pI. In OMJ-CIEF, the separation capillary is divided into seven equal sections joined with each other via tubular Nafion membrane insertions. Each junction is communicated with its own external electrolytic buffer which is used both to supply electrical contact and for solvent exchange. The performance of the fractionator was explored using protein and peptide samples covering broad pI range. Separation was achieved in ionic and ampholytic buffers, including ammonium formate, ammonium hydroxide, histidine, and arginine. By maintaining electric potential across upstream segments of the capillary after the focusing stage, selective release of downstream analyte fractions could be achieved. The selective release mode circumvents the problem of peak broadening during mobilization and enables convenient comprehensive sampling for orthogonal separation methods. Using single-component ampholyte buffers with well-defined pI cutoff values, controlled separation of protein mixture into basic and acidic fractions was demonstrated. The device is cheap and easy to fabricate in-house, simple in operation, and straightforward in interfacing to hyphened analytical platforms. OMJ-CIEF has a potential of becoming a practical add-on unit in a wide range of bioanalytical setups, in particular as a first-dimension separation in mass spectrometry based proteomics or as a preparative tool for analyte purification, fractionation, and preconcentration.


Gadocoletic acid trisodium salt (b22956/1): a new blood pool magnetic resonance contrast agent with application in coronary angiography.
Inversion recovery, three-dimensional, gradient-recalled echo magnetic resonance coronary angiography (IR-3D-GRE-MRCA), performed after administration of an intravascular T1-relaxing agent with prolonged permanence in the blood, is one of the most promising approaches to noninvasive magnetic resonance imaging (MRI) of the coronaries. The aim of the present study was the evaluation of the physicochemical properties in solution, pharmacokinetics, elimination from the body, protein binding, and signal enhancement characteristics of gadocoletic acid trisodium salt (B22956/1), a candidate gadolinium-based MRI contrast agent for coronary angiography. The pharmacokinetics and elimination from the body of gadocoletate ion, the contrastographically active component of gadocoletic acid trisodium salt, was evaluated after intravenous administration in rats and monkeys, using for assays high-performance liquid chromatography, x-ray fluorescence, and inductively coupled plasma atomic emission spectrometry. The binding of the gadocoletate ion to animal and human serum albumin was studied by means of ultrafiltration. The imaging properties of blood outside coronary arteries after contrast agent administration were evaluated in cynomolgus monkeys (Macaca fascicularis) by measuring aortic signal-to-noise and contrast-to-noise ratios in lower body angiograms. The suitability of gadocoletic acid trisodium salt for achieving contrast-enhanced magnetic resonance coronary angiography (ceMRCA) was tested in Yucatan micropigs with an IR-3D-GRE sequence. All in vivo relaxation rate measurement and images were obtained using a 1.5 T Siemens Symphony scanner. The fractional binding of gadocoletate ion at a concentration of 0.5 mM to serum albumin at the physiological concentration was 95%, 92%, 88%, and 86% for human, monkey, pig, and rat, respectively. In rats and monkeys, gadocoletate ion was excreted unmetabolized through the biliary and urinary routes. It was recovered with feces depending on the injected dose in percentages from 18% to 97%, providing evidence for a saturable biliary pathway. Plasma pharmacokinetics showed the complete elimination of gadocoletate ion within 24 hours after administration. In the monkey, the gadocoletate ion showed the pharmacokinetic behavior of a compound with partial vascular confinement and long plasma half-life, which may be ascribed to elevated binding to serum albumin. These properties manifested themselves in lower body angiograms with excellent image contrast between vessels and muscle. The slowly decaying aortic blood signal-to-noise and contrast-to-noise ratios over a 15-minute period is expected to allow 3-dimensional coronary angiography. The potential of gadocoletic acid trisodium salt for ceMRCA was also demonstrated in Yucatan micropigs. Elevated blood signal intensity and almost total myocardial signal suppression was maintained for almost 1 hour after administration, ie, for much longer than expected to be necessary for coronary angiography. During the whole period high resolution images of the right coronary artery could be obtained. On the basis of the pharmacokinetic profile and imaging characteristics, gadocoletic acid trisodium salt shows promise as a MR contrast agent for coronary angiography.


A cluster of methylations in the domain IV of 25S rRNA is required for ribosome stability.
In all three domains of life ribosomal RNAs are extensively modified at functionally important sites of the ribosome. These modifications are believed to fine-tune the ribosome structure for optimal translation. However, the precise mechanistic effect of modifications on ribosome function remains largely unknown. Here we show that a cluster of methylated nucleotides in domain IV of 25S rRNA is critical for integrity of the large ribosomal subunit. We identified the elusive cytosine-5 methyltransferase for C2278 in yeast as Rcm1 and found that a combined loss of cytosine-5 methylation at C2278 and ribose methylation at G2288 caused dramatic ribosome instability, resulting in loss of 60S ribosomal subunits. Structural and biochemical analyses revealed that this instability was caused by changes in the structure of 25S rRNA and a consequent loss of multiple ribosomal proteins from the large ribosomal subunit. Our data demonstrate that individual RNA modifications can strongly affect structure of large ribonucleoprotein complexes.


Reflective practice in nursing care: embedded assumptions in qualitative studies.
Qualitative nursing researchers have long recognized that reflective practice (RP) seems to be a valuable tool in nursing care. The aim of the present meta-study was to analyse current qualitative research on RP in nursing care, in order to create and synthesize the knowledge and the understanding of registered nurses' RP. Using a meta-study synthesis approach, embedded assumptions were identified in qualitative studies that have influenced the way researchers have interpreted and made sense of RP in nursing care. Despite empirical focus in research on RP in nursing care, it was found that assumptions about RP were predominantly based on theory. The reflective movement within the practice of nursing care has mainly a constructivist epistemology, based on learning from experience. The individual nurse's RP capability is essential in providing and improving ethical and holistic nursing care.


Size, velocity, and concentration in suspension measurements of spherical droplets and cylindrical jets.
The principle of an optical technique for simultaneous velocity, size, and concentration in suspension measurements of spherical droplets and cylindrical jets is proposed. This technique is based on phase Doppler anemometry working in the dual burst technique configuration. The particle size and velocity are deduced from the reflected signal phase and frequency, whereas the amplitude ratio between the refracted and the reflected signals is used for measuring the concentration of small scatterers inside the particles. Numerical simulations, based on geometrical optics and a Monte Carlo model, and an experimental validation test on cylindrical jets made of various suspensions, are used to validate the principle of the proposed technique. It is believed that this new technique could be useful in investigating processes in which liquid suspensions are sprayed for surface coating, drying, or combustion applications.


Building sustainable operational research capacity in Pakistan: starting with tuberculosis and expanding to other public health problems.
Background: For many years, operational research capacity has been a challenge and has remained a low priority for the health sector in Pakistan. Building research capacity for developing a critical mass of researchers in Pakistan was done through Structured Operational Research and Training Initiative (SORT IT) courses in Paris and Asia between 2010 and 2016. Objective: The aim of this paper is to describe the journey of SORT-IT in Pakistan from its inception to progressive expansion and discuss the challenges and ways forward. Methods: The journey began with the training of the Pakistan NTP research team lead in 2010 in an international SORT IT course at Paris. This was followed by training of two team members in Asia SORT IT courses in 2014 and 2015. These three then worked together to conceive and implement the first national Pakistan SORT IT course supported by WHO/TDR and the Global Fund in 2016. This was facilitated by international facilitators and local trained SORT-IT participants from Paris and Asia. This was followed by two further national SORT IT courses in 2017 and 2018. Results: Between 2010 and 2017, a total of 34 participants from Pakistan had been enrolled in national and international SORT IT courses. Of the 23 participants from completed courses, 18(78%) successfully completed the course. In total 18 papers were submitted and up until June 2018, 15(83%) have been published and 21 institutions in Pakistan involved with operational research as a result of the SORT IT initiative. Conclusions: The SORT IT course has been an effective way to build operational research capacity at national level and this has resulted in a large number of published papers providing local evidence for decision making on TB and other disease control programmes. The experience from Pakistan should stimulate other countries to adopt the SORT-IT model.


KRAS Mutant Pancreatic Cancer: No Lone Path to an Effective Treatment.
Pancreatic ductal adenocarcinoma (PDAC) is among the deadliest cancers with a dismal 7% 5-year survival rate and is projected to become the second leading cause of cancer-related deaths by 2020. KRAS is mutated in 95% of PDACs and is a well-validated driver of PDAC growth and maintenance. However, despite comprehensive efforts, an effective anti-RAS drug has yet to reach the clinic. Different paths to inhibiting RAS signaling are currently under investigation in the hope of finding a successful treatment. Recently, direct RAS binding molecules have been discovered, challenging the perception that RAS is an "undruggable" protein. Other strategies currently being pursued take an indirect approach, targeting proteins that facilitate RAS membrane association or downstream effector signaling. Unbiased genetic screens have identified synthetic lethal interactors of mutant RAS. Most recently, metabolic targets in pathways related to glycolytic signaling, glutamine utilization, autophagy, and macropinocytosis are also being explored. Harnessing the patient's immune system to fight their cancer is an additional exciting route that is being considered. The "best" path to inhibiting KRAS has yet to be determined, with each having promise as well as potential pitfalls. We will summarize the state-of-the-art for each direction, focusing on efforts directed toward the development of therapeutics for pancreatic cancer patients with mutated KRAS.


Controlled-reflectance surfaces with film-coupled colloidal nanoantennas.
Efficient and tunable absorption is essential for a variety of applications, such as designing controlled-emissivity surfaces for thermophotovoltaic devices, tailoring an infrared spectrum for controlled thermal dissipation and producing detector elements for imaging. Metamaterials based on metallic elements are particularly efficient as absorbing media, because both the electrical and the magnetic properties of a metamaterial can be tuned by structured design. So far, metamaterial absorbers in the infrared or visible range have been fabricated using lithographically patterned metallic structures, making them inherently difficult to produce over large areas and hence reducing their applicability. Here we demonstrate a simple method to create a metamaterial absorber by randomly adsorbing chemically synthesized silver nanocubes onto a nanoscale-thick polymer spacer layer on a gold film, making no effort to control the spatial arrangement of the cubes on the film. We show that the film-coupled nanocubes provide a reflectance spectrum that can be tailored by varying the geometry (the size of the cubes and/or the thickness of the spacer). Each nanocube is the optical analogue of a grounded patch antenna, with a nearly identical local field structure that is modified by the plasmonic response of the metal's dielectric function, and with an anomalously large absorption efficiency that can be partly attributed to an interferometric effect. The absorptivity of large surface areas can be controlled using this method, at scales out of reach of lithographic approaches (such as electron-beam lithography) that are otherwise required to manipulate matter on the nanoscale.


"Topical nephropathy" and "tropical extramembranous glomerulonephritis" of unknown aetiology in Senegal.
A study of renal biopsy specimens obtained in Senegal from 24 children and six adults with nephrotic syndrome showed two unusual varieties of nephropathy--namely, an extramembranous glomerulonephritis associated with hypocomplementaemia (four cases), a combination previously described only in systemic lupus erythematosus, and a "tropical nephropathy" (16 cases). The latter, though lacking the diffuse glomerular deposits of immunoglobulin described in quartan malarial nephropathy (Q.M.N.), showed a curious progressive and segmental glomerulosclerosis, characterized by a "flaking" or fibrillary splitting of the glomerular capillary wall, seen in Q.M.N. Serological evidence of malaria was lacking in a third of the childhood cases.


Physical and mental determinants of dropout and retention among nursing students: protocol of the SPRiNG cohort study.
The shortage of nursing professionals is of growing concern. The causes of this include the demanding physical and mental workload, leading to a dropout of nurses that may start during their education. However, it is unclear to what extent nursing students already perceive a physical and mental workload leading to health problems during their nursing education and placement, and to what extent these health problems cause students to dropout from nursing education. Very few prospective cohort studies have investigated protective and risk factors in relation to dropout and retention among nursing students. Three cohorts of third-year nursing students will be followed for 2.5 years. Students will be enrolled from the Bachelor of Nursing program of the Rotterdam University of Applied Sciences. At baseline, students will receive a self-administered questionnaire. Primary outcome is dropout from nursing education and dropout from the nursing profession. Data on dropout from nursing education will be retrieved from the student administration on a yearly basis. Dropout from the nursing profession will be measured one year after graduation, using the self-reported questionnaire. Secondary outcomes are presenteeism and sick leave (during internship/work). In addition to student characteristics, the questionnaire asks about physical and mental internship/work characteristics, personal and behavioral factors, and experienced physical and mental burden.Main aims of this study are to determine: 1) the prevalence and incidence rates of dropout, 2) the protective and risk factors, and early indicators of dropout, and 3) the interaction between these factors and the indicators. Data analysis of a large, prospective cohort study with regard to determinants of dropout and retention of nursing students and newly graduated nurses is in progress. Findings emerging from this study can be used to develop a predictive model to identify the first indicators of dropout from nursing education and nursing profession, for which targeted interventions can be deployed.


Risk of Spontaneous Abortion After Inadvertent Human Papillomavirus Vaccination in Pregnancy.
To evaluate the risk of spontaneous abortion after quadrivalent human papillomavirus (4vHPV) vaccination before and during pregnancy across seven integrated health systems within the Vaccine Safety Datalink. Within a retrospective observational cohort, we compared risks for spontaneous abortion after 4vHPV in three exposure windows: distal (16-22 weeks before the last menstrual period [LMP]), peripregnancy (within 6 weeks before the LMP), and during pregnancy (LMP through 19 weeks of gestation). Women 12-27 years of age with a pregnancy between 2008 and 2014, with continuous insurance enrollment 8 months before and through pregnancy end, and with a live birth, stillbirth, or spontaneous abortion were included. Pregnancies were identified through validated algorithms. Spontaneous abortions and stillbirths were verified by chart review with spontaneous abortions adjudicated by clinical experts. We excluded multiple gestations, spontaneous abortions before 6 weeks of gestation, and women using medications increasing risk of spontaneous abortion. Spontaneous abortion risk after 4vHPV during pregnancy was compared with distal vaccination using time-dependent covariate Cox models. Spontaneous abortion risk for peripregnancy compared with distal vaccination was evaluated with standard Cox models. We identified 2,800 pregnancies with 4vHPV exposure in specified risk windows: 919 (33%) distal, 986 (35%) peripregnancy, and 895 (32%) during pregnancy. Mean age was 22.4 years in distal and peripregnancy groups compared with 21.4 years among women vaccinated during pregnancy. Among women with distal 4vHPV exposure, 96 (10.4%) experienced a spontaneous abortion. For peripregnancy and during pregnancy exposures, spontaneous abortions occurred in 110 (11.2%) and 77 (8.6%), respectively. The risk of spontaneous abortion was not increased among women who received 4vHPV during pregnancy (adjusted hazard ratio 1.10, 95% CI 0.81-1.51) or peripregnancy 1.07 (0.81-1.41). Inadvertent 4vHPV exposure during or peripregnancy was not significantly associated with an increased risk of spontaneous abortion.


The Transplant Nephrology Workforce in the United States: Current State and Future Directions.
The population of patients with kidney transplants in the United States is growing. The delivery of transplant care is complex, involves a multidisciplinary transplant team, and care coordination between transplant and community providers. The transplant nephrologist is central to the delivery of this care and assumes a multitude of clinical and nonclinical roles and responsibilities. With a growing population of patients requiring transplant care that spans a continuum from pretransplant referral to long-term posttransplant management, an understanding of the current state of the transplant nephrology workforce in the United States and the future that it faces is important in ensuring that current and future needs of both patients and physicians are met. In this article, we (1) review the scope of practice of the transplant nephrologist, (2) discuss the state of training in the field of transplant nephrology, (3) review the role of the referring primary nephrologist in the care of patients undergoing kidney transplant, and (4) discuss challenges and opportunities facing the transplant nephrology workforce.


A national audit of the laboratory diagnosis of tuberculosis and other mycobacterial diseases within the United Kingdom.
In order to audit United Kingdom laboratory diagnostic and reference services including novel molecular methods for tuberculosis, a questionnaire was sent to laboratories submitting specimens to the PHLS Mycobacterium Reference Unit (MRU) and regional centres and to the Scottish Mycobacteria Reference Laboratory (SMRL) in 1996-7. Nationally, 67.2% of laboratories responded. Most UK laboratories were fully or conditionally CPA accredited and take part in the NEQAS proficiency scheme. On average only 3.3% of primary samples submitted for mycobacterial diagnosis in 1995 produced a mycobacterial culture from approximately half as many patients (that is, a mean of 1488 specimens producing 49 isolates from 23 patients). Potentially over 380,000 specimens are processed for mycobacteria in the UK each year. The majority of laboratories use 4% NaOH +/- NALC for specimen decontamination. Culture on solid media was used by most laboratories and 62.9% also use liquid media. Most laboratories incubated cultures for eight weeks. Few laboratories use molecular diagnostic methods. Laboratories were most likely to use molecular methods for diagnosing tuberculous meningitis and for specimens from immunocompromised patients, although usage was strongly influenced by cost. Within England and Wales 43.9% (47/107) and 56% (61/109) of laboratories wanted a rapid service for rifampicin resistance detection in M tuberculosis from immunocompetent and immunocompromised patients, respectively. In regard to a tuberculous meningitis service, 80.5% (43/112) and 84.3% (102/121) of laboratories wanted this service for immunocompetent and immunocompromised patients, respectively. The quality of reference services was rated as "very good"/"good" by 85.6% of respondents nationally. Rapid molecular amplification diagnostic services were established at the PHLS MRU for rifampicin drug resistance detection nationally and for tuberculous meningitis at the MRU.


Deletion of an amino-terminal sequence beta-catenin in vivo and promotes hyperphosporylation of the adenomatous polyposis coli tumor suppressor protein.
Regulation of cell adhesion and cell signaling by beta-catenin occurs through a mechanism likely involving the targeted degradation of the protein. Deletional analysis was used to generate a beta-catenin refractory to rapid turnover and to examine its effects on complexes containing either cadherin or the adenomatous polyposis coli (APC) protein. The results show that amino-terminal deletion of beta-catenin results in a protein with increased stability that acts in a dominant fashion with respect to wild-type beta-catenin. Constitutive expression in AtT20 cells of a beta-catenin lacking 89 N-terminal amino acids (deltaN89beta-catenin) resulted in severely reduced levels of the more labile wild-type beta-catenin. The mutant beta-catenin was expressed at endogenous levels but displaced the vast majority of wild-type beta-catenin associated with N-cadherin. The deltaN89beta-catenin accumulated on the APC protein to a level 10-fold over that of wild-type beta-catenin and recruited a kinase into the APC complex. The kinase was highly active toward APC in vitro and promoted a sodium dodecyl sulfate gel band shift that was also evident for endogenous APC from cells expressing the mutant beta-catenin. Unlike wild-type beta-catenin, which partitions solely as part of a high-molecular-weight complex, the deltaN89 mutant protein also fractionated as a stable monomer, indicating that it had escaped the requirement to associate with other proteins. That similar N-terminal mutants of beta-catenin have been implicated in cellular transformation suggests that their abnormal association with APC may, in part, be responsible for this phenotype.


Elective surgery system strengthening: development, measurement, and validation of the surgical preparedness index across 1632 hospitals in 119 countries.
The 2015 Lancet Commission on global surgery identified surgery and anaesthesia as indispensable parts of holistic health-care systems. However, COVID-19 exposed the fragility of planned surgical services around the world, which have also been neglected in pandemic recovery planning. This study aimed to develop and validate a novel index to support local elective surgical system strengthening and address growing backlogs. First, we performed an international consultation through a four-stage consensus process to develop a multidomain index for hospital-level assessment (surgical preparedness index; SPI). Second, we measured surgical preparedness across a global network of hospitals in high-income countries (HICs), middle-income countries (MICs), and low-income countries (LICs) to explore the distribution of the SPI at national, subnational, and hospital levels. Finally, using COVID-19 as an example of an external system shock, we compared hospitals' SPI to their planned surgical volume ratio (SVR; ie, operations for which the decision for surgery was made before hospital admission), calculated as the ratio of the observed surgical volume over a 1-month assessment period between June 6 and Aug 5, 2021, against the expected surgical volume based on hospital administrative data from the same period in 2019 (ie, a pre-pandemic baseline). A linear mixed-effects regression model was used to determine the effect of increasing SPI score. In the first phase, from a longlist of 103 candidate indicators, 23 were prioritised as core indicators of elective surgical system preparedness by 69 clinicians (23 [33%] women; 46 [67%] men; 41 from HICs, 22 from MICs, and six from LICs) from 32 countries. The multidomain SPI included 11 indicators on facilities and consumables, two on staffing, two on prioritisation, and eight on systems. Hospitals were scored from 23 (least prepared) to 115 points (most prepared). In the second phase, surgical preparedness was measured in 1632 hospitals by 4714 clinicians from 119 countries. 745 (45·6%) of 1632 hospitals were in MICs or LICs. The mean SPI score was 84·5 (95% CI 84·1-84·9), which varied between HIC (88·5 [89·0-88·0]), MIC (81·8 [82·5-81·1]), and LIC (66·8 [64·9-68·7]) settings. In the third phase, 1217 (74·6%) hospitals did not maintain their expected SVR during the COVID-19 pandemic, of which 625 (51·4%) were from HIC, 538 (44·2%) from MIC, and 54 (4·4%) from LIC settings. In the mixed-effects model, a 10-point increase in SPI corresponded to a 3·6% (95% CI 3·0-4·1; p<0·0001) increase in SVR. This was consistent in HIC (4·8% [4·1-5·5]; p<0·0001), MIC (2·8 [2·0-3·7]; p<0·0001), and LIC (3·8 [1·3-6·7%]; p<0·0001) settings. The SPI contains 23 indicators that are globally applicable, relevant across different system stressors, vary at a subnational level, and are collectable by front-line teams. In the case study of COVID-19, a higher SPI was associated with an increased planned surgical volume ratio independent of country income status, COVID-19 burden, and hospital type. Hospitals should perform annual self-assessment of their surgical preparedness to identify areas that can be improved, create resilience in local surgical systems, and upscale capacity to address elective surgery backlogs. National Institute for Health Research (NIHR) Global Health Research Unit on Global Surgery, NIHR Academy, Association of Coloproctology of Great Britain and Ireland, Bowel Research UK, British Association of Surgical Oncology, British Gynaecological Cancer Society, and Medtronic.


Hospital use of agency-employed supplemental nurses and patient mortality and failure to rescue.
To determine the association between the use of agency-employed supplemental registered nurses (SRNs) to staff hospitals and patient mortality and failure to rescue (FTR). Primary survey data from 40,356 registered nurses in 665 hospitals in four states in 2006 were linked with American Hospital Association and inpatient mortality data from state agencies for approximately 1.3 million patients. Logistic regression models were used to examine the association between SRN use and 30-day in-hospital mortality and FTR, controlling for patient and hospital characteristics, nurse staffing, the proportion of nurses with bachelor's degrees, and quality of the work environment. Before controlling for multiple nurse characteristics of hospitals, higher proportions of agency-employed SRNs in hospitals appeared to be associated with higher mortality (OR = 1.06) and FTR (OR = 1.05). Hospitals with higher proportions of SRNs have poorer work environments, however, and the significant relationships between SRNs and mortality outcomes were rendered insignificant when work environments were taken into account. Higher use of SRNs does not appear to have deleterious consequences for patient mortality and may alleviate nurse staffing problems that could produce higher mortality.


Alcohol-associated intestinal dysbiosis impairs pulmonary host defense against Klebsiella pneumoniae.
Chronic alcohol consumption perturbs the normal intestinal microbial communities (dysbiosis). To investigate the relationship between alcohol-mediated dysbiosis and pulmonary host defense we developed a fecal adoptive transfer model, which allows us to investigate the impact of alcohol-induced gut dysbiosis on host immune response to an infectious challenge at a distal organ, independent of prevailing alcohol use. Male C57BL/6 mice were treated with a cocktail of antibiotics (ampicillin, gentamicin, neomycin, vancomycin, and metronidazole) via daily gavage for two weeks. A separate group of animals was fed a chronic alcohol (or isocaloric dextrose pair-fed controls) liquid diet for 10 days. Microbiota-depleted mice were recolonized with intestinal microbiota from alcohol-fed or pair-fed (control) animals. Following recolonization groups of mice were sacrificed prior to and 48 hrs. post respiratory infection with Klebsiella pneumoniae. Klebsiella lung burden, lung immunology and inflammation, as well as intestinal immunology, inflammation, and barrier damage were examined. Results showed that alcohol-associated susceptibility to K. pneumoniae is, in part, mediated by gut dysbiosis, as alcohol-naïve animals recolonized with a microbiota isolated from alcohol-fed mice had an increased respiratory burden of K. pneumoniae compared to mice recolonized with a control microbiota. The increased susceptibility in alcohol-dysbiosis recolonized animals was associated with an increase in pulmonary inflammatory cytokines, and a decrease in the number of CD4+ and CD8+ T-cells in the lung following Klebsiella infection but an increase in T-cell counts in the intestinal tract following Klebsiella infection, suggesting intestinal T-cell sequestration as a factor in impaired lung host defense. Mice recolonized with an alcohol-dysbiotic microbiota also had increased intestinal damage as measured by increased levels of serum intestinal fatty acid binding protein. Collectively, these results suggest that alterations in the intestinal immune response as a consequence of alcohol-induced dysbiosis contribute to increased host susceptibility to Klebsiella pneumonia.


Excited-State Dynamics of 5,14- vs 6,13-Bis(trialkylsilylethynyl)-Substituted Pentacenes: Implications for Singlet Fission.
Singlet fission is a process in conjugated organic materials that has the potential to considerably improve the performance of devices in many applications, including solar energy conversion. In any application involving singlet fission, efficient triplet harvesting is essential. At present, not much is known about molecular packing arrangements detrimental to singlet fission. In this work, we report a molecular packing arrangement in crystalline films of 5,14-bis(triisopropylsilylethynyl)-substituted pentacene, specifically a local (pairwise) packing arrangement, responsible for complete quenching of triplet pairs generated via singlet fission. We first demonstrate that the energetic condition necessary for singlet fission is satisfied in amorphous films of the 5,14-substituted pentacene derivative. However, while triplet pairs form highly efficiently in the amorphous films, only a modest yield of independent triplets is observed. In crystalline films, triplet pairs also form highly efficiently, although independent triplets are not observed because triplet pairs decay rapidly and are quenched completely. We assign the quenching to a rapid nonadiabatic transition directly to the ground state. Detrimental quenching is observed in crystalline films of two additional 5,14-bis(trialkylsilylethynyl)-substituted pentacenes with either ethyl or isobutyl substituents. Developing a better understanding of the losses identified in this work, and associated molecular packing, may benefit overcoming losses in solids of other singlet fission materials.


Expanded circulating hematopoietic stem/progenitor cells as novel cell source for the treatment of TCIRG1 osteopetrosis.
Allogeneic hematopoietic stem cell transplantation is the treatment of choice for autosomal recessive osteopetrosis caused by defects in the TCIRG1 gene. Despite recent progress in conditioning, a relevant number of patients are not eligible for allogeneic stem cell transplantation because of the severity of the disease and significant transplant-related morbidity. We exploited peripheral CD34+ cells, known to circulate at high frequency in the peripheral blood of TCIRG1-deficient patients, as a novel cell source for autologous transplantation of gene corrected cells. Detailed phenotypical analysis showed that circulating CD34+ cells have a cellular composition that resembles bone marrow, supporting their use in gene therapy protocols. Transcriptomic profile revealed enrichment in genes expressed by hematopoietic stem and progenitor cells (HSPCs). To overcome the limit of bone marrow harvest/ HSPC mobilization and serial blood drawings in TCIRG1 patients, we applied UM171-based ex-vivo expansion of HSPCs coupled with lentiviral gene transfer. Circulating CD34+ cells from TCIRG1-defective patients were transduced with a clinically-optimized lentiviral vector (LV) expressing TCIRG1 under the control of phosphoglycerate promoter and expanded ex vivo. Expanded cells maintained long-term engraftment capacity and multi-lineage repopulating potential when transplanted in vivo both in primary and secondary NSG recipients. Moreover, when CD34+ cells were differentiated in vitro, genetically corrected osteoclasts resorbed the bone efficiently. Overall, we provide evidence that expansion of circulating HSPCs coupled to gene therapy can overcome the limit of stem cell harvest in osteopetrotic patients, thus opening the way to future gene-based treatment of skeletal diseases caused by bone marrow fibrosis.


Recruitment of Rpd3 to the telomere depends on the protein arginine methyltransferase Hmt1.
In the yeast Saccharomyces cerevisiae, the establishment and maintenance of silent chromatin at the telomere requires a delicate balance between opposing activities of histone modifying enzymes. Previously, we demonstrated that the protein arginine methyltransferase Hmt1 plays a role in the formation of yeast silent chromatin. To better understand the nature of the Hmt1 interactions that contribute to this phenomenon, we carried out a systematic reverse genetic screen using a null allele of HMT1 and the synthetic genetic array (SGA) methodology. This screen revealed interactions between HMT1 and genes encoding components of the histone deacetylase complex Rpd3L (large). A double mutant carrying both RPD3 and HMT1 deletions display increased telomeric silencing and Sir2 occupancy at the telomeric boundary regions, when comparing to a single mutant carrying Hmt1-deletion only. However, the dual rpd3/hmt1-null mutant behaves like the rpd3-null single mutant with respect to silencing behavior, indicating that RPD3 is epistatic to HMT1. Mutants lacking either Hmt1 or its catalytic activity display an increase in the recruitment of histone deacetylase Rpd3 to the telomeric boundary regions. Moreover, in such loss-of-function mutants the levels of acetylated H4K5, which is a substrate of Rpd3, are altered at the telomeric boundary regions. In contrast, the level of acetylated H4K16, a target of the histone deacetylase Sir2, was increased in these regions. Interestingly, mutants lacking either Rpd3 or Sir2 display various levels of reduction in dimethylated H4R3 at these telomeric boundary regions. Together, these data provide insight into the mechanism whereby Hmt1 promotes the proper establishment and maintenance of silent chromatin at the telomeres.


Allowing for non-adherence to treatment in a randomized controlled trial of two antidepressants (citalopram versus reboxetine): an example from the GENPOD trial.
Meta-analyses suggest that reboxetine may be less effective than other antidepressants. Such comparisons may be biased by lower adherence to reboxetine and subsequent handling of missing outcome data. This study illustrates how to adjust for differential non-adherence and hence derive an unbiased estimate of the efficacy of reboxetine compared with citalopram in primary care patients with depression. A structural mean modelling (SMM) approach was used to generate adherence-adjusted estimates of the efficacy of reboxetine compared with citalopram using GENetic and clinical Predictors Of treatment response in Depression (GENPOD) trial data. Intention-to-treat (ITT) analyses were performed to compare estimates of effectiveness with results from previous meta-analyses. At 6 weeks, 92% of those randomized to citalopram were still taking their medication, compared with 72% of those randomized to reboxetine. In ITT analysis, there was only weak evidence that those on reboxetine had a slightly worse outcome than those on citalopram [adjusted difference in mean Beck Depression Inventory (BDI) scores: 1.19, 95% confidence interval (CI) -0.52 to 2.90, p = 0.17]. There was no evidence of a difference in efficacy when differential non-adherence was accounted for using the SMM approach for mean BDI (-0.29, 95% CI -3.04 to 2.46, p = 0.84) or the other mental health outcomes. There was no evidence of a difference in the efficacy of reboxetine and citalopram when these drugs are taken and tolerated by depressed patients. The SMM approach can be implemented in standard statistical software to adjust for differential non-adherence and generate unbiased estimates of treatment efficacy for comparisons of two (or more) active interventions.


The evolution of cranial design and performance in squamates: Consequences of skull-bone reduction on feeding behavior.
The evolution of cranial design in lepidosaurians is characterized by a general trend toward the loss of cranial elements. The evolution of relatively lighter skulls in squamates appears tightly coupled to a reduction in relative mass of the jaw adductor, implying functional consequences for bite force and feeding behavior. Interestingly, among squamates the postorbital bar was reduced or lost at least twice independently and taxa characterized by the loss of these cranial elements (e.g., geckos and varanids) are generally reported as having a mobile skull. In Gekkotans, the loss of the postorbital bar was accompanied by a reduction of the supratemporal bar, resulting in a pronounced cranial kinesis. Our data show that having a kinetic skull has functional consequences and results in a reduction in bite force. The lower bite force may in turn be responsible for the decreased feeding efficiency as reflected in the longer duration of intra-oral transport cycles. Gekkotans, however, appear to exploit their intracranial mobility in ways that increases the velocity of jaw movement during opening and closing, which may allow them to capture more elusive prey. The morphological changes observed in the evolution of the cranial system in squamates appear tightly linked to functional and constructional demands on the skull, making squamate skull evolution a model system to investigate the consequences of morphological changes in a complex integrated system of performance, behavior, and ecology.


A descriptive study of reportable gastrointestinal illnesses in Ontario, Canada, from 2007 to 2009.
Gastrointestinal illnesses (GI) continue to pose a substantial burden in terms of morbidity and economic impact in Canada. We describe the epidemiology of reportable GI in Ontario by characterizing the incidence of each reportable GI, as well as associated demographics, clinical outcomes, seasonality, risk settings, and likely sources of infection. Reports on laboratory confirmed cases of amebiasis, botulism, campylobacteriosis, cryptosporidiosis, cyclosporiasis, giardiasis, hepatitis A, listeriosis, paratyphoid fever, salmonellosis, shigellosis, typhoid fever, illness due to verotoxin-producing Escherichia coli (VTEC-illness), and yersiniosis, from January 1, 2007 to December 31, 2009 were obtained from Ontario's passive reportable disease surveillance system. Cases were classified by history of relevant travel, association with outbreaks, and likely source of infection, obtained through follow-up of reported cases by local health authorities. There were 29,897 GI reported by health authorities in Ontario from 2007 to 2009. The most frequently reported diseases were campylobacteriosis (10,916 cases or 36.5% of all GI illnesses) and salmonellosis (7,514 cases, 25.1%). Overall, 26.9% of GI cases reported travel outside of Ontario during the relevant incubation period. Children four years of age and younger had the highest incidence rate for most GI, and significantly more (54.8%, p<0.001) cases occurred among males than females. The most commonly reported sources of infections were food (54.2%), animals (19.8%), and contact with ill persons (16.9%). Private homes (45.5%) and food premises (29.7%) were the most commonly reported exposure settings. Domestic cases of campylobacteriosis, cryptosporidiosis, giardiasis, salmonellosis, and VTEC-illness showed seasonal patterns with incidence peaking in the summer months. Reportable GI continues to be a burden in Ontario. Since more than one in four GI cases experienced in Ontario were acquired outside of the province, international travel is an important risk factor for most GI. Because private homes are the most commonly reported risk settings and the main suspect sources of infection are food, animal contact and ill persons, these findings support the continued need for public health food safety programs, public education on safe handling of food and animals, and proper hand hygiene practices.


Impact of dengue virus infection on feeding behavior of Aedes aegypti.
In addition to heavily infecting the salivary glands of Aedes aegypti (L.) mosquitoes, dengue viruses produce a significant infection of the nervous system, involving the brain, Johnston's organ, compound eye, and thoracic and abdominal ganglion. To determine if dengue infection affects feeding behavior of Ae. aegypti we measured feeding times, counted the number of feeding delays or interruptions, and by in situ immunocytochemistry techniques determined the spatial and temporal distribution of dengue infections in females parenterally infected with dengue 3 virus. The mean of the total time required for feeding by infected mosquitoes was significantly longer than the time required by uninfected mosquitoes. Similarly, the mean of the time spent probing was significantly longer in infected mosquitoes than in uninfected mosquitoes when day after inoculation was considered. Significant increases in the length of feeding activity in infected mosquitoes corresponded to virus infection in organs that are known to control or influence activities associated with blood feeding. Sequential infections of the salivary glands (five days postinoculation [PI]), brain and compound eye (eight days PI), and Johnston's organ and midgut and abdominal ganglion (11 days PI) of most mosquitoes were observed. The increased time required by infected Ae. aegypti mosquitoes to acquire a blood meal may contribute to the efficiency of Ae. aegypti as a vector of dengue virus. Longer feeding periods are more likely to be interrupted by the host, which increases the chance that an infected mosquito will probe or feed on additional hosts.


Patient satisfaction and costs associated with insulin administered by pen device or syringe during hospitalization.
Patient satisfaction, safety and efficacy outcomes, and cost savings with insulin pens versus conventional insulin delivery via vials and syringes in hospitalized patients with diabetes were compared. Patients were recruited from two general medical-surgical units from July 2005 to May 2006. Patients completed a survey regarding satisfaction with the method in which insulin was administered before discharge. Patients completed a telephone survey approximately four weeks after discharge to determine home insulin use. Cost savings were determined using the average wholesale price of insulin vials and syringes, pens, and pen needles. A total of 94 patients were randomized to receive insulin administered via pen devices (n = 49) or using conventional vials and syringes (n = 45). Significantly more subjects in the pen group prepared or self-injected at least one dose of insulin during hospitalization, wanted to continue taking insulin at home using the method used during hospitalization, and would recommend their method of insulin administration used during hospitalization to other patients with diabetes compared with the vial and syringe group (p < 0.05). A cost saving of $36 per patient was projected if only insulin pens were dispensed during the entire hospital stay compared to insulin vials and syringes (p < 0.05). Increased patient satisfaction and continuation of the method of insulin administration used in the hospital at home were reported by patients who received insulin pens compared with patients who received conventional vials and syringes during hospitalization. A substantial cost saving was projected for patients in the insulin pen group if insulin pens had been dispensed during their entire hospital stay.


Modelling of diffraction grating based optical filters for fluorescence detection of biomolecules.
The detection of biomolecules based on fluorescence measurements is a powerful diagnostic tool for the acquisition of genetic, proteomic and cellular information. One key performance limiting factor remains the integrated optical filter, which is designed to reject strong excitation light while transmitting weak emission (fluorescent) light to the photodetector. Conventional filters have several disadvantages. For instance absorbing filters, like those made from amorphous silicon carbide, exhibit low rejection ratios, especially in the case of small Stokes' shift fluorophores (e.g. green fluorescent protein GFP with λ exc = 480 nm and λ em = 510 nm), whereas interference filters comprising many layers require complex fabrication. This paper describes an alternative solution based on dielectric diffraction gratings. These filters are not only highly efficient but require a smaller number of manufacturing steps. Using FEM-based optical modelling as a design optimization tool, three filtering concepts are explored: (i) a diffraction grating fabricated on the surface of an absorbing filter, (ii) a diffraction grating embedded in a host material with a low refractive index, and (iii) a combination of an embedded grating and an absorbing filter. Both concepts involving an embedded grating show high rejection ratios (over 100,000) for the case of GFP, but also high sensitivity to manufacturing errors and variations in the incident angle of the excitation light. Despite this, simulations show that a 60 times improvement in the rejection ratio relative to a conventional flat absorbing filter can be obtained using an optimized embedded diffraction grating fabricated on top of an absorbing filter.