Influenza risk management: lessons learned from an A(H1N1) pdm09 outbreak investigation in an operational military setting.
At the onset of an influenza pandemic, when the severity of a novel strain is still undetermined and there is a threat of introduction into a new environment, e.g., via the deployment of military troops, sensitive screening criteria and conservative isolation practices are generally recommended. In response to elevated rates of influenza-like illness among U.S. military base camps in Kuwait, U.S. Naval Medical Research Unit No. 3 partnered with local U.S. Army medical units to conduct an A(H1N1) pdm09 outbreak investigation. Initial clinical data and nasal specimens were collected via the existent passive surveillance system and active surveillance was conducted using a modified version of the World Health Organization/U.S. Centers for Disease Control and Prevention influenza-like illness case definition [fever (T > 100.5˚F/38˚C) in addition to cough and/or sore throat in the previous 72 hours] as the screening criteria. Samples were tested via real-time reverse-transcription PCR and sequenced for comparison to global A(H1N1) pdm09 viruses from the same time period. The screening criteria used in Kuwait proved insensitive, capturing only 16% of A(H1N1) pdm09-positive individuals. While still not ideal, using cough as the sole screening criteria would have increased sensitivity to 73%. The results of and lessons learned from this outbreak investigation suggest that pandemic influenza risk management should be a dynamic process (as information becomes available regarding true attack rates and associated mortality, screening and isolation criteria should be re-evaluated and revised as appropriate), and that military operational environments present unique challenges to influenza surveillance.


Molecularly imprinted polymer coated on stainless steel fiber for solid-phase microextraction of chloroacetanilide herbicides in soybean and corn.
A molecularly imprinted polymer (MIP) with metolachlor as template was firstly coated on stainless steel fiber through chemical bonding strategy to solve the fragility problem of silica fiber substrate for solid-phase microextraction. The surface pretreatment of stainless steel fiber and the polymerization conditions were investigated systematically to enhance the preparation feasibility and MIP coating performance, and then a porous and highly cross-linked MIP coating with 14.8-microm thickness was obtained with over 200 times re-usability which was supported by non-fragile stainless steel fiber adoption. The MIP coating possessed specific selectivities to metolachlor, its metabolites and other chloroacetanilide herbicides with the factors of 1.1-4.6. Good extraction capacities of metolachlor, propisochlor and butachlor were found with MIP coating under quick adsorption and desorption kinetics, and the detection limits of 3.0, 9.6 and 38 microg L(-1) were achieved, respectively. Moreover, the MIP-coated stainless steel fiber was evaluated for trace metolachlor, propisochlor and butachlor extraction in the spiked soybean and corn samples, and the enrichment factors of 54-60, 27-31 and 15-20 were obtained, respectively.


IQ in late adolescence/early adulthood, risk factors in middle age and later all-cause mortality in men: the Vietnam Experience Study.
To examine the role of potential mediating factors in explaining the IQ-mortality relation. A total of 4316 male former Vietnam-era US army personnel with IQ test results at entry into the service in late adolescence/early adulthood in the 1960/1970s (mean age at entry 20.4 years) participated in a telephone survey and medical examination in middle age (mean age 38.3 years) in 1985-6. They were then followed up for mortality experience for 15 years. In age-adjusted analyses, higher IQ scores were associated with reduced rates of total mortality (hazard ratio (HR)(per SD increase in IQ) 0.71; 95% CI 0.63 to 0.81). This relation did not appear to be heavily confounded by early socioeconomic position or ethnicity. The impact of adjusting for some potentially mediating risk indices measured in middle age on the IQ-mortality relation (marital status, alcohol consumption, systolic and diastolic blood pressure, pulse rate, blood glucose, body mass index, psychiatric and somatic illness at medical examination) was negligible (<10% attenuation in risk). Controlling for others (cigarette smoking, lung function) had a modest impact (10-17%). Education (0.79; 0.69 to 0.92), occupational prestige (0.77; 0.68 to 0.88) and income (0.86; 0.75 to 0.98) yielded the greatest attenuation in the IQ-mortality gradient (21-52%); after their collective adjustment, the IQ-mortality link was effectively eliminated (0.92; 0.79 to 1.07). In this cohort, socioeconomic position in middle age might lie on the pathway linking earlier IQ with later mortality risk but might also partly act as a surrogate for cognitive ability.


Secondary prevention of variceal bleeding in adults with previous oesophageal variceal bleeding due to decompensated liver cirrhosis: a network meta-analysis.
Approximately 40% to 95% of people with cirrhosis have oesophageal varices. About 15% to 20% of oesophageal varices bleed in about one to three years of diagnosis. Several different treatments are available, which include endoscopic sclerotherapy, variceal band ligation, beta-blockers, transjugular intrahepatic portosystemic shunt (TIPS), and surgical portocaval shunts, among others. However, there is uncertainty surrounding their individual and relative benefits and harms. To compare the benefits and harms of different initial treatments for secondary prevention of variceal bleeding in adults with previous oesophageal variceal bleeding due to decompensated liver cirrhosis through a network meta-analysis and to generate rankings of the different treatments for secondary prevention according to their safety and efficacy. We searched CENTRAL, MEDLINE, Embase, Science Citation Index Expanded, World Health Organization International Clinical Trials Registry Platform, and trials registers until December 2019 to identify randomised clinical trials in people with cirrhosis and a previous history of bleeding from oesophageal varices. We included only randomised clinical trials (irrespective of language, blinding, or status) in adults with cirrhosis and previous history of bleeding from oesophageal varices. We excluded randomised clinical trials in which participants had no previous history of bleeding from oesophageal varices, previous history of bleeding only from gastric varices, those who failed previous treatment (refractory bleeding), those who had acute bleeding at the time of treatment, and those who had previously undergone liver transplantation. We performed a network meta-analysis with OpenBUGS using Bayesian methods and calculated the differences in treatments using hazard ratios (HR), odds ratios (OR) and rate ratios with 95% credible intervals (CrI) based on an available-case analysis, according to National Institute of Health and Care Excellence Decision Support Unit guidance. We included a total of 48 randomised clinical trials (3526 participants) in the review. Forty-six trials (3442 participants) were included in one or more comparisons. The trials that provided the information included people with cirrhosis due to varied aetiologies. The follow-up ranged from two months to 61 months. All the trials were at high risk of bias. A total of 12 interventions were compared in these trials (sclerotherapy, beta-blockers, variceal band ligation, beta-blockers plus sclerotherapy, no active intervention, TIPS (transjugular intrahepatic portosystemic shunt), beta-blockers plus nitrates, portocaval shunt, sclerotherapy plus variceal band ligation, beta-blockers plus nitrates plus variceal band ligation, beta-blockers plus variceal band ligation, sclerotherapy plus nitrates). Overall, 22.5% of the trial participants who received the reference treatment (chosen because this was the commonest treatment compared in the trials) of sclerotherapy died during the follow-up period ranging from two months to 61 months. There was considerable uncertainty in the effects of interventions on mortality. Accordingly, none of the interventions showed superiority over another. None of the trials reported health-related quality of life. Based on low-certainty evidence, variceal band ligation may result in fewer serious adverse events (number of people) than sclerotherapy (OR 0.19; 95% CrI 0.06 to 0.54; 1 trial; 100 participants). Based on low or very low-certainty evidence, the adverse events (number of participants) and adverse events (number of events) may be different across many comparisons; however, these differences are due to very small trials at high risk of bias showing large differences in some comparisons leading to many differences despite absence of direct evidence. Based on low-certainty evidence, TIPS may result in large decrease in symptomatic rebleed than variceal band ligation (HR 0.12; 95% CrI 0.03 to 0.41; 1 trial; 58 participants). Based on moderate-certainty evidence, any variceal rebleed was probably lower in sclerotherapy than in no active intervention (HR 0.62; 95% CrI 0.35 to 0.99, direct comparison HR 0.66; 95% CrI 0.11 to 3.13; 3 trials; 296 participants), beta-blockers plus sclerotherapy than sclerotherapy alone (HR 0.60; 95% CrI 0.37 to 0.95; direct comparison HR 0.50; 95% CrI 0.07 to 2.96; 4 trials; 231 participants); TIPS than sclerotherapy (HR 0.18; 95% CrI 0.08 to 0.38; direct comparison HR 0.22; 95% CrI 0.01 to 7.51; 2 trials; 109 participants), and in portocaval shunt than sclerotherapy (HR 0.21; 95% CrI 0.05 to 0.77; no direct comparison) groups. Based on low-certainty evidence, beta-blockers alone and TIPS might result in more, other compensation, events than sclerotherapy (rate ratio 2.37; 95% CrI 1.35 to 4.67; 1 trial; 65 participants and rate ratio 2.30; 95% CrI 1.20 to 4.65; 2 trials; 109 participants; low-certainty evidence). The evidence indicates considerable uncertainty about the effect of the interventions including those related to beta-blockers plus variceal band ligation in the remaining comparisons. The evidence indicates considerable uncertainty about the effect of the interventions on mortality. Variceal band ligation might result in fewer serious adverse events than sclerotherapy. TIPS might result in a large decrease in symptomatic rebleed than variceal band ligation. Sclerotherapy probably results in fewer 'any' variceal rebleeding than no active intervention. Beta-blockers plus sclerotherapy and TIPS probably result in fewer 'any' variceal rebleeding than sclerotherapy. Beta-blockers alone and TIPS might result in more other compensation events than sclerotherapy. The evidence indicates considerable uncertainty about the effect of the interventions in the remaining comparisons. Accordingly, high-quality randomised comparative clinical trials are needed.


Inverse relationship between intensity of vaginal self-stimulation-produced analgesia and level of chronic intake of a dietary source of capsaicin.
Women who chronically ingest a diet rich in capsaicin, the pungent ingredient in hot chili peppers, showed a significantly lower magnitude of analgesia in response to vaginal self-stimulation than women with relatively low or medium levels of ingestion. Vaginal self-stimulation-produced analgesia was quantified by measuring (on the hand) pain detection thresholds, pain tolerance thresholds and tactile thresholds. Whereas vaginal self-stimulation produced a 32.6-43.8% increase in pain detection and pain tolerance thresholds in the low chili diet group, it produced only a 2.3-7.3% increase in these measures in the high chili diet group. The medium chili diet group showed an intermediate effect on the pain thresholds. Tactile thresholds were not increased by the vaginal self-stimulation. Baseline (no stimulation) pain thresholds did not differ significantly among the three groups. These findings are consistent with earlier studies in laboratory rats, in which capsaicin administered neonatally abolished vaginal stimulation-produced analgesia, but did not affect baseline pain thresholds to mechanostimulation.


Is external carotid endarterectomy a durable procedure?
Thirteen patients with internal carotid occlusion underwent 14 primary external carotid revascularization procedures over a 31 month period. Ten patients had obliteration of their internal carotid stump combined with patch angioplasty of the external carotid artery, and 3 had vein bypasses from the common carotid artery to the external carotid artery. Eleven patients were symptomatic with either amaurosis fugax or hemispheric transient ischemic attacks. Two patients were asymptomatic. All patients had serial carotid noninvasive tests (B-mode ultrasonography, spectral analysis, and oculoplethysmography). The mean follow-up was 22 months. Recurrent amaurosis fugax secondary to recurrent stenoses developed in two patients. These were correctly predicted by B-mode imaging and altered flow characteristics on spectral analysis. Both patients were successfully treated with reoperative procedures to prevent failure of the primary reconstruction. External carotid revascularization is a safe and durable procedure, but careful periodic follow-up is necessary to detect stenoses developing at or remote from the initial operative site. Carotid noninvasive tests appear to be helpful in detecting recurrent disease. Carotid revascularization is superior to other forms of therapy in patients who have development of neurologic symptoms ipsilateral to a chronically occluded internal carotid artery.


Mechanism of inhibition of HIV-1 reverse transcriptase by non-nucleoside inhibitors.
The structure of unliganded HIV-1 reverse transcriptase has been determined at 2.35 A resolution and refined to an R-factor of 0.219 (for all data) with good stereochemistry. The unliganded structure was produced by soaking out a weak binding non-nucleoside inhibitor, HEPT, from pregrown crystals. Comparison with the structures of four different RT and non-nucleoside inhibitor complexes reveals that only minor domain rearrangements occur, but there is a significant repositioning of a three-stranded beta-sheet in the p66 subunit (containing the catalytic aspartic acid residues 110, 185 and 186) with respect to the rest of the polymerase site. This suggests that NNIs inhibit RT by locking the polymerase active site in an inactive conformation, reminiscent of the conformation observed in the inactive p51 subunit.


Assessing the efficacy, effectiveness, and cost-effectiveness of assistive technology interventions for enhancing mobility.
The aim of this paper is to highlight the contributions that complementary efficacy, effectiveness, and cost-effectiveness studies can make to assessing the outcomes of assistive technology interventions for enhancing mobility. The terms, 'assistive technology outcomes research' and 'assistive technology interventions', are defined. Several bases are examined for the shortage of outcomes research pertaining to mobility-related assistive technology interventions. Three presuppositions are described for the research strategy of interlocking studies being recommended. They are assigning priority to evaluating both recently developed assistive technologies and ones that have long been available, acknowledging the complexity of assistive technology as an intervention, and appreciating the trade-offs necessary for strengthening studies' internal and external validity. Some key study preparations are considered, including treatment theory, treatment specification, and the selection of outcome domains and measures. The essential features of efficacy, effectiveness, and cost-effectiveness studies are outlined, and their interdependence is stressed. To assess the outcomes of assistive technology interventions for mobility in ways that are both methodologically sound and relevant to stakeholder needs, a research strategy is required involving mutually reinforcing efficacy, effectiveness, and cost-effectiveness studies. Collaborative arrangements and funding methods are discussed for fostering the needed research.


Short turn radius colonoscope in an anatomical model: retroflexed withdrawal and detection of hidden polyps.
To evaluate the new RetroView™ colonoscope and compare its ability to detect simulated polyps "hidden" behind colonic folds with that of a conventional colonoscope, utilizing anatomic colon models. Three anatomic colon models were prepared, with twelve simulated polyps "hidden" behind haustral folds and five placed in easily viewed locations in each model. Five blinded endoscopists examined two colon models in random order with the conventional or RetroView™ colonoscope, utilizing standard withdrawal technique. The third colon model was then examined with the RetroView™ colonoscope withdrawn initially in retroflexion and then in standard withdrawal. Polyp detection rates during standard and retroflexed withdrawal of the conventional and RetroView™ colonoscopes were determined. Polyp detection rates for combined standard and retroflexed withdrawal (combination withdrawal) with the RetroView™ colonoscope were also determined. For hidden polyps, retroflexed withdrawal using the RetroView™ colonoscope detected more polyps than the conventional colonoscope in standard withdrawal (85% vs 12%, P = 0.0001). For hidden polyps, combination withdrawal with the RetroView™ colonoscope detected more polyps than the conventional colonoscope in standard withdrawal (93% vs 12%, P ≤ 0.0001). The RetroView™ colonoscope in "combination withdrawal" was superior to other methods in detecting all (hidden + easily visible) polyps, with successful detection of 80 of 85 polyps (94%) compared to 28 (32%) polyps detected by the conventional colonoscope in standard withdrawal (P < 0.0001) and 67 (79%) polyps detected by the RetroView™ colonoscope in retroflexed withdrawal alone (P < 0.01). Continuous withdrawal of the colonoscope through the colon model while retroflexed was achieved by all endoscopists. In a post-test survey, four out of five colonoscopists reported that manipulation of the colonoscope was easy or very easy. In simulated testing, the RetroView™ colonoscope increased detection of hidden polyps. Combining standard withdrawal with retroflexed withdrawal may become the new paradigm for "complete screening colonoscopy".


RGS16 attenuates pulmonary Th2/Th17 inflammatory responses.
The regulators of G protein signaling (RGS) protein superfamily negatively controls G protein-coupled receptor signal transduction pathways. RGS16 is enriched in activated/effector T lymphocytes. In this paper, we show that RGS16 constrains pulmonary inflammation by regulating chemokine-induced T cell trafficking in response to challenge with Schistosoma mansoni. Naive Rgs16(-/-) mice were "primed" for inflammation by accumulation of CCR10(+) T cells in the lung. Upon pathogen exposure, these mice developed more robust granulomatous lung fibrosis than wild-type counterparts. Distinct Th2 or putative Th17 subsets expressing CCR4 or CCR10 accumulated more rapidly in Rgs16(-/-) lungs following challenge and produced proinflammatory cytokines IL-13 and IL-17B. CCR4(+)Rgs16(-/-) Th2 cells migrated excessively to CCL17 and localized aberrantly in challenged lungs. T lymphocytes were partially excluded from lung granulomas in Rgs16(-/-) mice, instead forming peribronchial/perivascular aggregates. Thus, RGS16-mediated confinement of T cells to Schistosome granulomas mitigates widespread cytokine-mediated pulmonary inflammation.


Functional responses and prey-stage preferences of a predatory gall midge and two predacious mites with twospotted spider mites, Tetranychus urticae, as host.
The twospotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), is an important pest of vegetables and other economically important crops. This study evaluated the functional responses and prey-stage preferences of three species of predators, a predatory gall midge, Feltiella acarisuga (Vallot) (Diptera: Cecidomyiidae), and two predatory mite species, Neoseiulus californicus (McGregor) (Acari: Phytoseiidae) and Amblyseius swirskii (AnthiasHenriot), with T. urticae as the host, under laboratory conditions. The results showed that F. acarisuga was highly effective and the two species of predacious mites were moderately effective in feeding on T. urticae eggs. Logistic regression analysis suggested Type II (convex) functional responses for all three species. However, based on the estimates of the handling time and the attacking rates, the three predators had different predation capacities. Among the three species, F. acarisuga had the highest predation on T. urticae. The maximum daily predation by a larval F. acarisuga was 50 eggs/day, followed by a female N. californicus (25.6 eggs/day) and a female A. swirskii (15.1 eggs/day). A female N. californicus produced more eggs than a female A. swirskii did when they both fed on T. urticae eggs. In addition, all three predator species had no preystage preference for either prey eggs or nymphs. The findings from this study could help select better biological control agents for effective control of T. urticae and other pests in vegetable productions.


Development of a new promoter to avoid the silencing of genes in the production of recombinant antibodies in chinese hamster ovary cells.
The production of recombinant proteins in mammalian cell lines is one of the most important areas in biopharmaceutical industry. Viral transcriptional promoters are widely used to express recombinant proteins in mammalian cell lines. However, these promoters are susceptible to silencing, thus limiting protein productivity. Some CpG islands can avoid the silencing of housekeeping genes; for that reason, they have been used to increase the production of recombinant genes in cells of animal origin. In this study, we evaluated the CpG island of the promoter region of the β-actin gene of Cricetulus griseous (Chinese hamster), associated to the Cytomegalovirus (CMV) promoter, to increase recombinant antibodies production in Chinese Hamster Ovary (CHO) cells. We focused on the non-coding region of CpG island, which we called RegCG. RegCG behaved as a promoter, whose transcriptional activity was mainly commanded by the CAAT and CArG boxes of the proximal promoter. However, the transcription started mainly at the intronic region before the proximal transcription start site. While the CMV promoter was initially more powerful than RegCG, the latter promoter was more resistant to silencing than the CMV promoter in stable cell lines, and its activity was improved when combined with the CMV promoter. Thereby, the chimeric promoter was able to maintain the expression of recombinant antibodies in stable clones for 40 days at an average level 4 times higher than the CMV promoter. Finally, the chimeric promoter showed compatibility with a genetic amplification system by induction with methotrexate in cells deficient in the dihydrofolate reductase gene. We have generated an efficient synthetic hybrid transcription promoter through the combination of RegCG with CMV, which, in stable cell lines, shows greater activity than when both promoters are used separately. Our chimeric promoter is compatible with a genetic amplification system in CHO DG44 cells and makes possible the generation of stable cell lines with high production of recombinant antibodies. We propose that this promoter can be a good alternative for the generation of clones expressing high amount of recombinant proteins, essential for industrial applications.


Prospective evaluation of mannan and anti-mannan antibodies for diagnosis of invasive Candida infections in patients with neutropenic fever.
The diagnostic performance and usefulness of the Platelia antigen and antibody test (Bio-Rad) was investigated in a prospective study of haematological patients at risk for invasive Candida infections. Among 100 patients, 86 were eligible, of whom invasive candidiasis (IC) occurred in 12 (14%), according to the criteria of the European Organization for Research and Treatment of Cancer/Mycoses Study Group. These included candidaemia due to Candida albicans (one patient) or Candida tropicalis (four patients), and hepatosplenic candidiasis (seven patients). The comparator group of 74 patients included 50 with febrile neutropenia alone and 24 with mould infections. A strategy was developed to determine diagnostic cut-offs from receiver operating characteristic curves with maximal sensitivity and, given this sensitivity, maximal specificity, both being greater than 0. In this patient population, these values were 0.25 ng ml(-1) for mannan (M) and 2.6 arbitrary units ml(-1) for anti-mannan (AM), which are lower than those recommended by the manufacturer. All patients developed at least one positive diagnostic M or AM result during the 10 days of persistent febrile neutropenia (PFN). The optimal overall performance was found when two consecutive positive tests for both M and AM were used [sensitivity, specificity, positive predictive value and negative predictive value (NPV) (95 % confidence intervals) of 0.73 (0.39-0.94), 0.80 (0.69-0.89), 0.36 (0.17-0.59) and 0.95 (0.86-0.99), respectively]. There was a positive correlation of M with beta-D-glucan (r=0.28, P=0.01). The first positive M test was found up to a mean+/-sd of 8.8+/-8.5 (range 2-23) days prior to a clinical/mycological diagnosis of IC. Day-to-day variation in quantitative M levels was high. High-level AM responses were delayed until leucopenia resolved. The low specificities of the test performance may have been due to some of the comparator patients having subclinical Candida infections as evidenced by the high incidence of colonization among them (60% had a colonization index of >or=0.5). The high NPVs suggest that the tests may be particularly useful in excluding IC. It is feasible to explore the use of serial measurements of M and AM as part of a broader diagnostic strategy for selecting PFN patients to receive antifungal drug therapy.


Intra-operative erythropoietin during laparoscopic partial nephrectomy is not renoprotective.
In pre-clinical studies, acute erythropoietin (EPO) administration has been shown to mitigate the deleterious effects of ischemia/reperfusion injury. We reviewed our clinical experience with intraoperative EPO administration as a potential renoprotective agent during laparoscopic partial nephrectomy (LPN). Patients who underwent LPN at our institution between August 2008 and March 2010 received 500 IU/kg EPO 30 min prior to hilar occlusion. Those who underwent LPN between August 2006 and July 2008 without receiving EPO were selected as controls. Demographic, clinical, perioperative, and estimated glomerular filtration rate (eGFR) data were compared for the cohorts preoperatively, and during short-term (<6 months) and long-term (≥6 months) follow-up. Short-term eGFR was evaluable for 39 EPO and 29 controls, while long-term eGFR was evaluable for 26 EPO and 27 controls. Baseline demographic and clinical features of the cohorts were similar. For EPO versus controls, median short and long-term follow-up was 19 days versus 22 days and 10.2 months versus 11.9 months, respectively. Mean preoperative, postoperative, and % change in eGFR were statistically similar for the cohorts during short- and long-term follow-up, without and with adjustment for baseline renal function (unadjusted P-values = 0.28, 0.095, and 0.38, respectively, short term, and 0.61, 0.50, and 0.69, respectively, long term). In this retrospective study, a single dose of EPO prior to hilar occlusion during LPN had no added protective impact on postoperative eGFR in the short or long term. Prospective evaluation in patients with solitary kidneys may better elucidate its potential renoprotective role in this setting.


Prevalence and correlates of depression among veterans with multiple sclerosis.
To establish the prevalence of major depressive episode (MDE) in a large sample of veterans with multiple sclerosis (MS); to identify demographic characteristics, aspects of disease presentation, and perceptions of disability associated with greater concurrent risk for MDE; and to examine the relationship between MDE, service utilization, and activity participation. Veterans with MS (n = 1,032) were identified via computer database and surveyed by mail; 451 (43.7%) responded. Twenty-two percent of the sample met criteria for current MDE. Low income, unemployment, presence of falls, younger age, absence of a marital partner, and high levels of perceived disability due to bowel functioning were independently associated with MDE. Disease subtype, disease duration, use of disease modifying therapies, and perceived disability due to mobility or bladder problems were unrelated to MDE. Current MDE was in turn associated with increased primary care visits and increased impact of disease upon activity participation. Similar correlates were associated with minor depressive episode. Unlike the general population, rates of depression in this predominantly male sample were similar to those found in predominantly female samples of persons with multiple sclerosis. Specific aspects of disability were differentially associated with depression, and depression was independently associated with increased service utilization and increased participation limitations.


Using register data to estimate causal effects of interventions: An ex post synthetic control-group approach.
It is common in the context of evaluations that participants have not been selected on the basis of transparent participation criteria, and researchers and evaluators many times have to make do with observational data to estimate effects of job training programs and similar interventions. The techniques developed by researchers in such endeavours are useful not only to researchers narrowly focused on evaluations, but also to social and population science more generally, as observational data overwhelmingly are the norm, and the endogeneity challenges encountered in the estimation of causal effects with such data are not trivial. The aim of this article is to illustrate how register data can be used strategically to evaluate programs and interventions and to estimate causal effects of participation in these. We use propensity score matching on pretreatment-period variables to derive a synthetic control group, and we use this group as a comparison to estimate the employment-treatment effect of participation in a large job-training program. We find the effect of treatment to be small and positive but transient. Our method reveals a strong regression to the mean effect, extremely easy to interpret as a treatment effect had a less advanced design been used (e.g. a within-subjects panel data analysis), and illustrates one of the unique advantages of using population register data for research purposes.


Representing properties locally.
Theories of knowledge such as feature lists, semantic networks, and localist neural nets typically use a single global symbol to represent a property that occurs in multiple concepts. Thus, a global symbol represents mane across HORSE, PONY, and LION. Alternatively, perceptual theories of knowledge, as well as distributed representational systems, assume that properties take different local forms in different concepts. Thus, different local forms of mane exist for HORSE, PONY, and LION, each capturing the specific form that mane takes in its respective concept. Three experiments used the property verification task to assess whether properties are represented globally or locally (e.g., Does a PONY have mane?). If a single global form represents a property, then verifying it in any concept should increase its accessibility and speed its verification later in any other concept. Verifying mane for PONY should benefit as much from having verified mane for LION earlier as from verifying mane for HORSE. If properties are represented locally, however, verifying a property should only benefit from verifying a similar form earlier. Verifying mane for PONY should only benefit from verifying mane for HORSE, not from verifying mane for LION. Findings from three experiments strongly supported local property representation and ruled out the interpretation that object similarity was responsible (e.g., the greater overall similarity between HORSE and PONY than between LION and PONY). The findings further suggest that property representation and verification are complicated phenomena, grounded in sensory-motor simulations.


In silico Docking Studies of Fingolimod and S1P1 Agonists.
The sphingosine-1-phosphate receptor 1 (S1P1), originally the endothelial differentiation gene 1 receptor (EDG-1), is one of five G protein-coupled receptors (GPCRs) S1P1 - 5 that bind to and are activated by sphingosine-1-phosphate (S1P). The lipid S1P is an intermediate in sphingolipid homeostasis, and S1P1 is a major medical target for immune system modulation; agonism of the receptor produces a myriad of biological responses, including endothelial cell barrier integrity, chemotaxis, lymphocyte trafficking/targeting, angiogenesis, as well as regulation of the cardiovascular system. Use of in silico docking simulations on the crystal structure of S1P1 allows for pinpointing the residues within the receptor's active site that actively contribute to the binding of S1P, and point to how these specific interactions can be exploited to design more effective synthetic analogs to specifically target S1P1 in the presence of the closely related receptors S1P2, S1P3, S1P4, and S1P5. We examined the binding properties of the endogenous substrate as well as a selection of synthetic sphingosine-derived S1P1 modulators of S1P1 with in silico docking simulations using the software package Molecular Operating Environment® (MOE®). The modeling studies reveal the relevance of phosphorylation, i.e., the presence of a phosphate or phosphonate moiety within the substrate for successful binding to occur, and indicate which residues are responsible for S1P1 binding of the most prominent sphingosine-1-phosphate receptor (S1PR) modulators, including fingolimod and its structural relatives. Furthermore, trends in steric preferences as for the binding of enantiomers to S1P1 could be observed, facilitating future design of receptor-specific substrates to precisely target the active site of S1P1.


Losartan reverses impaired osseointegration in spontaneously hypertensive rats.
Hypertension is not only associated with cardiovascular diseases but also with alterations in bone quality. Hypertension therefore might be a risk factor for osseointegration. Preclinical studies suggest that losartan, an angiotensin II receptor blocker widely used to treat hypertension, has a beneficial effect in graft consolidation. Here, we determine the effect of hypertension and losartan on osseointegration. We used spontaneously hypertensive rats (SHR) and normotensive Wistar albinus rats receiving losartan (30 mg/kg, p.o.) or left untreated. After 1 week, titanium miniscrews were inserted into the tibia. Sixty days after implantation, implant stability was evaluated by removal torque measurement considered the primary endpoint. Microcomputed tomography and histomorphometric analysis were secondary endpoints. Losartan increased the removal torque in the hypertensive SHR group to levels of the Wistar controls. While the cortical parameters of osseointegration remained unchanged, losartan increased medullary bone formation. Microcomputed tomography revealed a higher bone volume per tissue volume and trabecular thickness in the SHR rats treated with losartan. Histomorphometric analysis further showed that losartan significantly increased the thickness of newly formed bone in medullary area in hypertensive SHR rats. Losartan did not significantly alter the parameters of osseointegration in normotensive rats. The data presented suggest that the angiotensin II receptor antagonist losartan increases the medullary parameters of osseointegration in a tibia model of spontaneously hypertensive rats. Considering the study limitations, understanding the impact of hypertension and the respective drugs on osseointegration requires further research.


Color constancy of color reproductions in art paintings.
Popular color reproductions of art paintings such as postcards are intended to remind viewers of the original works. It is, however, unclear how well the quality of the reproductions is preserved under various illuminations. Color constancy of the reproductions in relation to colors in the original paintings was estimated computationally with hyperspectral images of 15th-century Flemish paintings, 20th-century modern abstract paintings, and their corresponding postcards with a series of illuminants: the CIE daylight D65 with correlated color temperature (CCT) 6500 K, daylight D40, fluorescent lamps F2 and F11, and a LED lamp designed for museums with CCT approximately 3500-4000 K. Despite large colorimetric differences between the types of art paintings and between the illuminants simulated, local areas showed good color constancy: skin areas in the Flemish paintings ranged from 0.76 to 0.81, whereas nonskin areas ranged from 0.19 to 0.68. This result suggests that viewers may be able to achieve color constancy with the reproduction postcards by disregarding inconsistent colors representations from the original paintings caused by changes in illumination conditions.


Curcumin mediated suppression of nuclear factor-κB promotes chondrogenic differentiation of mesenchymal stem cells in a high-density co-culture microenvironment.
Osteoarthritis (OA) and rheumatoid arthritis (RA) are characterised by joint inflammation and cartilage degradation. Although mesenchymal stem cell (MSC)-like progenitors are resident in the superficial zone of articular cartilage, damaged tissue does not possess the capacity for regeneration. The high levels of pro-inflammatory cytokines present in OA/RA joints may impede the chondrogenic differentiation of these progenitors. Interleukin (IL)-1β activates the transcription factor nuclear factor-κB (NF-κB), which in turn activates proteins involved in matrix degradation, inflammation and apoptosis. Curcumin is a phytochemical capable of inhibiting IL-1β-induced activation of NF-κB and expression of apoptotic and pro-inflammatory genes in chondrocytes. Therefore, the aim of the present study was to evaluate the influence of curcumin on IL-1β-induced NF-κB signalling pathway in MSCs during chondrogenic differentiation. MSCs were either cultured in a ratio of 1:1 with primary chondrocytes in high-density culture or cultured alone in monolayer with/without curcumin and/or IL-1β. We demonstrate that although curcumin alone does not have chondrogenic effects on MSCs, it inhibits IL-1β-induced activation of NF-κB, activation of caspase-3 and cyclooxygenase-2 in MSCs time and concentration dependently, as it does in chondrocytes. In IL-1β stimulated co-cultures, four-hour pre-treatment with curcumin significantly enhanced the production of collagen type II, cartilage specific proteoglycans (CSPGs), β1-integrin, as well as activating MAPKinase signaling and suppressing caspase-3 and cyclooxygenase-2. Curcumin treatment may help establish a microenvironment in which the effects of pro-inflammatory cytokines are antagonized, thus facilitating chondrogenesis of MSC-like progenitor cells in vivo. This strategy may support the regeneration of articular cartilage.


Plastic debris on Pacific Islands: Ecological and health implications.
Plastic debris is a worldwide problem. This is particularly acute in the Pacific region, where its scale is a reason for serious concerns. There is an obvious need for studies to assess the extent to which plastic debris affects the Pacific. Therefore, this research aims to address this need by undertaking a systematic assessment of the ecological and health impacts of plastic debris on Pacific islands. Using pertinent historical qualitative and quantitative data of the distribution of plastic debris in the region, this study identified pollution and contamination trends and risks to ecosystems, and suggests some measures which may be deployed to address the identified problems. The study illustrates the fact that Pacific Island States are being disproportionately affected by plastic, and reiterates that further studies and integrated strategies are needed, involving public education and empowerment, governmental action, as well as ecologically sustainable industry leadership. It is also clear that more research is needed in respect of developing alternatives to conventional plastic, by the production of bio-plastic, i.e. plastic which is produced from natural (e.g. non-fossil fuel-based sources) materials, and which can be fully biodegradable.


Cognitive function in an elderly population: interaction between vitamin B12 status, depression, and apolipoprotein E ε4: the Hordaland Homocysteine Study.
To investigate the cross-sectional relation between metabolic markers of vitamin B(12) status and cognitive performance, and possible effect modification by the presence of depression and apolipoprotein E (ApoE) ε4. This is a population-based study of 1935 participants, aged 71 to 74 years, from Norway. Participants were administered a cognitive test battery, and vitamin B(12) status was assessed by measurements of plasma vitamin B(12), holotranscobalamin (holoTC), methylmalonic acid (MMA), and total homocysteine. The geometric mean (95% confidence interval) for vitamin B(12) was 348 pM (341-354), whereas 5.9% of participants had vitamin B(12) levels lower than 200 pM. In linear regression analyses, holoTC (p = .039) and the holoTC/vitamin B(12) ratio (p = .013) were positively related, whereas MMA (p = .010) was inversely related, to global cognition, after adjustment for sex, education, ApoE status, plasma creatinine, and history of diabetes, cardiovascular disease, hypertension, and depression. Among those positive for ApoE ε4, but not among those without the ε4 allele, plasma vitamin B(12) was positively associated with global cognition (p = .015), whereas MMA was inversely related to global cognition (p = .036) and executive function (p = .014). In participants with depression, MMA was inversely associated with global cognition (p < .001) and episodic memory (p = .001). Among the well-nourished elderly, low vitamin B(12) status is associated with cognitive deficit, particularly in those with the ApoE ε4 allele or with depression.


Development and validation of Emotional Labour in Nursing Scale in Italy.
Emotional demands may be a significant risk factor for wellbeing in healthcare professionals. Nurses have been considered the most exposed to emotional burden known as emotional labour. Scholars define it as the process by which nurses sometimes hide/fake the real emotion or struggle to display the appropriate emotion to meet the emotional work requirements. Emotional labour can result in physical, psychological and behavioural disorders. A specific tool to measure it among nurses is still missing. This study aims to develop and validate an emotional labour scale for nurses. The design process consisted of three different phases. In the first one, 24 items have been selected from different tools. The second consisted of the assessment of the face and content validity. In the third phase, the Emotional labour scale has been psychometrically evaluated through a cross validation approach, including both exploratory factor analysis performed on a sample of 205 Italian nurses, with the final selection of an 11- items tool, and confirmatory factor analysis. Emotional labour is acted through surface acting, compliance and restraint. The Emotional Labour Nursing scale showed good psychometric properties and was found to be valid and reliable for assessing emotional labour among nurses. The association between emotional labour and burnout was investigated. The Emotional Labour scale may foster a fuller understanding of the consequences of emotional labour. Since emotional demands and emotional labour are not included in the most common psychosocial risk assessment tools currently available, our results point out to their role in work stress prevention.


Elevated CXC chemokines in urine noninvasively discriminate OAB from UTI.
Overlapping symptoms of overactive bladder (OAB) and urinary tract infection (UTI) often complicate the diagnosis and contribute to overprescription of antibiotics. Inflammatory response is a shared characteristic of both UTI and OAB and here we hypothesized that molecular differences in inflammatory response seen in urine can help discriminate OAB from UTI. Subjects in the age range of (20-88 yr) of either sex were recruited for this urine analysis study. Urine specimens were available from 62 UTI patients with positive dipstick test before antibiotic treatment. Six of these patients also provided urine after completion of antibiotic treatment. Subjects in cohorts of OAB (n = 59) and asymptomatic controls (n = 26) were negative for dipstick test. Urinary chemokines were measured by MILLIPLEX MAP Human Cytokine/Chemokine Immunoassay and their association with UTI and OAB was determined by univariate and multivariate statistics. Significant elevation of CXCL-1, CXCL-8 (IL-8), and CXCL-10 together with reduced levels for a receptor antagonist of IL-1A (sIL-1RA) were seen in UTI relative to OAB and asymptomatic controls. Elevated CXCL-1 urine levels predicted UTI with odds ratio of 1.018 and showed a specificity of 80.77% and sensitivity of 59.68%. Postantibiotic treatment, reduction was seen in all CXC chemokines with a significant reduction for CXCL-10. Strong association of CXCL-1 and CXCL-10 for UTI over OAB indicates mechanistic differences in signaling pathways driving inflammation secondary of infection in UTI compared with a lack of infection in OAB. Urinary chemokines highlight molecular differences in the paracrine signaling driving the overlapping symptoms of UTI and OAB.


Selective staining and eradication of cancer cells by protein-carrying DARPin-functionalized liposomes.
Since their discovery, liposomes have been widely employed in biomedical research. These nano-size spherical vesicles consisting one or few phospholipid bilayers surrounding an aqueous core are capable of carrying a wide variety of bioactive compounds, including drugs, peptides, nucleic acids, proteins and others. Despite considerable success achieved in synthesis of liposome constructs containing bioactive compounds, preparation of ligand-targeted liposomes comprising large quantities of encapsulated proteins that are capable of affecting pathological cells still remains a big challenge. Here we described a novel method for preparation of small (80-90 nm in diameter) unilamellar liposomes containing very large quantities (thousands of protein molecules per liposome) of heme-containing cytochrome c, highly fluorescent mCherry and highly toxic PE40 (Pseudomonas aeruginosa Exotoxin A domain). Efficient encapsulation of the proteins was achieved through electrostatic interaction between positively charged proteins (at pH lower than pI) and negatively charged liposome membrane. The proteoliposomes containing large quantities of mCherry or PE40 and functionalized with designed ankyrin repeat protein (DARPin)_9-29, which targets human epidermal growth factor receptor 2 (HER2) were shown to specifically stain and kill in sub-nanomolar concentrations HER2-positive cells, overexpressing HER2, respectively. Specific staining and eradication of the receptor-positive cells demonstrated here makes the DARPin-functionalized liposomes carrying large quantities of fluorescent and/or toxic proteins a promising candidate for tumor detection and therapy.


Mutation Update for Kabuki Syndrome Genes KMT2D and KDM6A and Further Delineation of X-Linked Kabuki Syndrome Subtype 2.
Kabuki syndrome (KS) is a rare but recognizable condition that consists of a characteristic face, short stature, various organ malformations, and a variable degree of intellectual disability. Mutations in KMT2D have been identified as the main cause for KS, whereas mutations in KDM6A are a much less frequent cause. Here, we report a mutation screening in a case series of 347 unpublished patients, in which we identified 12 novel KDM6A mutations (KS type 2) and 208 mutations in KMT2D (KS type 1), 132 of them novel. Two of the KDM6A mutations were maternally inherited and nine were shown to be de novo. We give an up-to-date overview of all published mutations for the two KS genes and point out possible mutation hot spots and strategies for molecular genetic testing. We also report the clinical details for 11 patients with KS type 2, summarize the published clinical information, specifically with a focus on the less well-defined X-linked KS type 2, and comment on phenotype-genotype correlations as well as sex-specific phenotypic differences. Finally, we also discuss a possible role of KDM6A in Kabuki-like Turner syndrome and report a mutation screening of KDM6C (UTY) in male KS patients.


MALDI imaging reveals lipid changes in the skin of leprosy patients before and after multidrug therapy (MDT).
Leprosy still represents a health problem in several countries. Affecting skin and peripheral nerves, it may lead to permanent disabilities. Disturbances on skin lipid metabolism in leprosy were already observed; however, the localization and distribution of lipids could not be accessed. The role of lipids on infectious disease has been fully addressed only recently, as they directly influence immune response. Matrix-assisted laser desorption/ionization imaging mass spectrometry provides a powerful tool to localize and identify lipids in tissues. The aim of this work was to study and compare the changes in lipid distribution of skin biopsies taken from leprosy patients before and after multidrug therapy (MDT). Different species of phosphatidic acid, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin and phosphatidylcholine were detected. Differences in skin lipid signal intensities, as well as in their localization, were observed before and after MDT in every patient. In general, lipid distribution in the skin after MDT had a pattern similar to control skin samples, where most of the lipids were located in the upper part of the dermis and epidermis. This study opens paths to a better understanding of lipid functions in leprosy pathogenesis and immune response.


Identification and localization of three classes of myosins in pollen tubes of Lilium longiflorum and Nicotiana alata.
The presence and localization of actin and myosin have been examined in pollen tubes of Lilium longiflorum and Nicotiana alata. Immunoblot analysis of pollen tube extracts with antibodies to actin, myosins IA and IB, myosin II, and myosin V reveals the presence of these contractile proteins. Immunofluorescence microscopy using various methods to preserve the pollen tubes; chemical fixation, rapid freeze fixation and freeze substitution (RF-FS) followed by rehydration or by embeddment in a methacrylate mixture, was performed to optimize preservation. Immunocytochemistry reaffirmed that actin is localized longitudinally in the active streaming lanes and near the cortical surface of the pollen tube. Myosin I was localized to the plasma membrane, larger organelles, the surface of the generative cell and the vegetative nucleus, whereas, myosin V was found in the vegetative cytoplasm in a punctate fashion representing smaller organelles. Myosin II subfragment 1 and light meromyosin were localized in a punctate fashion on the larger organelles throughout the vegetative cytoplasm. In addition, isolated generative cells and vegetative nuclei labeled only with the myosin I antibody. Competition studies indicated the specificity of the heterologous antibodies utilized in this study suggesting the presence of three classes of myosins in pollen. These results lead to the following hypothesis: Myosin I may move the generative cell and vegetative nucleus unidirectionally through the pollen tube to the tip, while myosin V moves the smaller organelles and myosins I and II move the larger organelles (bidirectionally) that are involved in growth.


Toward the identification, characterization and experimental culture of Lacazia loboi from Atlantic bottlenose dolphin (Tursiops truncatus).
Lobomycosis (lacaziosis) is a chronic, granulomatous, fungal infection of the skin and subcutaneous tissues of humans and dolphins. To date, the causative agent, the yeast-like organism Lacazia loboi, has not been grown in the laboratory, and there have been no recent reports describing attempts to culture the organism. As a result, studies on the efficacy of therapeutics and potential environmental reservoirs have not been conducted. Therefore, the objective of the current study was to utilize both classical and novel microbiological methods in order to stimulate growth of Lacazia cells collected from dolphin lesions. This included the experimental inoculation of novel media, cell culture, and the use of artificial skin matrices. Although unsuccessful, the methods and results of this study provide important insight into new approaches that could be utilized in future investigations of this elusive organism.


Genomic loci of the Porphyromonas gingivalis insertion element IS1126.
The Porphyromonas gingivalis genome contains multiple copies of insertion element IS1126. When chromosomal DNA digests of different strains were probed with IS1126, between 25 and 35 hybridizing fragments per genome were detected, depending on the strain. Unrelated strains had very different restriction fragment length polymorphism (RFLP) patterns. When different laboratory copies of a specific strain were examined, the IS1126 RFLP patterns were very similar but small differences were observed, indicating that element-associated changes had occurred during laboratory passage. Within the next year, genome sequencing, assembly, and annotation for P. gingivalis W83 will be completed. Because repetitive elements complicate the assembly of randomly sequenced DNA fragments, we isolated and sequenced the flanking regions of IS1126 copies in strain W83. We also isolated and sequenced the flanking regions of IS1126 copies in strain ATCC 33277 in order to compare insertion sites in phylogenetically divergent strains. We identified 37 new sequences flanking IS1126 from strain ATCC 33277 and 30 from strain W83. The insertion element was found between genes except where it transposed into another insertion element. Examination of identifiable flanking genes or open reading frames indicated that the insertion sites were different in the two strains, except that both strains possess an insertion adjacent to the Lys-gingipain gene (J. P. Lewis and F. L. Macrina, Infect. Immun. 66:3035-3042, 1998). Most of the genes or sequences flanking IS1126 in ATCC 33277 were present in W83 but were contiguous and not insertion element associated. Thus, where genes were identified in both strains, their order was maintained, indicating that the two genomes are organized similarly, but the loci of IS1126 are different. In both strains, insertion element-associated duplicated target sites were lost from several copies of IS1126, providing evidence of homologous recombination between elements. Larger organizational differences between the genomes, such as deletions and inversions, may result from insertion element-mediated recombination events.


Peripheral lymphoid hyperplasia and central lymphoid depletion in mice treated with a bacterial B-cell mitogen (F3'EP-Si/p90).
In order to further understand the mechanism mediating the mitogenic and immunosuppressor effects of p90, a protein produced by Streptococcus intermedius, flow cytometric studies were performed on peripheral and central lymphoid organs of mice treated with this protein. p90 induced a strong blastogenic B-cell response in the spleen and lymph nodes, followed by a slight but significant polyclonal T-cell activation. B-cell repertoire analysis indicated that polyclonal B-cell responses affected similarly both CD5+ and conventional (CD5-) B cells in the spleen. Repertoire analysis of T cells failed to reveal any preferential stimulation of the V beta T-cell receptor (V beta-TcR) families studied. Peripheral lymphoid hyperplasia was observed concomitantly with central lymphoid depletion. In the bone marrow, pre-B and B cells were profoundly depleted, with a more pronounced effect on small pre-B cells. In the thymus, double-positive (CD4+CD8+) thymocytes were preferentially eliminated, with a relative enrichment of single positive (either CD4+ or CD8+) and double-negative (CD4-CD8-) thymocytes.


Traversal of the Blood-Brain Barrier by Cleavable l-Lysine Conjugates of Apigenin.
Apigenin, a flavone abundant in parsley and celery, is known to act on several CNS receptors, but its very poor water solubility (<0.001 mg/mL) impedes its absorption in vivo and prevents clinical use. Herein, apigenin was directly conjugated with glycine, l-phenylalanine, and l-lysine to give the corresponding carbamate derivatives, all of which were much more soluble than apigenin itself (0.017, 0.018, and 0.13 mg/mL, respectively). The Lys-apigenin carbamate 10 had a temporary sedative effect on the mice within 5 min of intraperitoneal administration (single dose of 0.4 mg/g) and could be detected in the mice brain tissues at a concentration of 0.82 μg/g of intact Lys-apigenin carbamate 10 and 0.42 ug/g of apigenin at 1.5 h. This study accomplished the delivery of apigenin across the BBB in a manner that might be applicable to other congeners, which should inform the future development of BBB-crossing flavonoids.


Particle alignment and clustering in sheared granular materials composed of platy particles.
By means of molecular dynamics simulations, we investigate the texture and local ordering in sheared packings composed of cohesionless platy particles. The morphology of large packings of platy particles in quasistatic equilibrium is complex due to the combined effects of local nematic ordering of the particles and anisotropic orientations of contacts between particles. We find that particle alignment is strongly enhanced by the degree of platyness and leads to the formation of face-connected clusters of exponentially decaying size. Interestingly, due to dynamics in continuous shearing, this ordering phenomenon emerges even in systems composed of particles of very low platyness differing only slightly from spherical shape. The number of clusters is an increasing function of platyness. However, at high platyness the proportion of face-face interactions is too low to allow for their percolation throughout the system.


Impaired nitric oxide production in coronary endothelial cells of the spontaneously diabetic BB rat is due to tetrahydrobiopterin deficiency.
Endothelial cells (EC) from diabetic BioBreeding (BB) rats have an impaired ability to produce NO. This deficiency is not due to a defect in the constitutive isoform of NO synthase in EC (ecNOS) or alterations in intracellular calcium, calmodulin, NADPH or arginine levels. Instead, ecNOS cannot produce sufficient NO because of a deficiency in tetrahydrobiopterin (BH(4)), a cofactor necessary for enzyme activity. EC from diabetic rats exhibited only 12% of the BH(4) levels found in EC from normal animals or diabetes-prone animals which did not develop disease. As a result, NO synthesis by EC of diabetic rats was only 18% of that for normal animals. Increasing BH(4) levels with sepiapterin increased NO production, suggesting that BH(4) deficiency is a metabolic basis for impaired endothelial NO synthesis in diabetic BB rats. This deficiency is due to decreased activity of GTP-cyclohydrolase I, the first and rate-limiting enzyme in the de novo biosynthesis of BH(4). GTP-cyclohydrolase activity was low because of a decreased expression of the protein in the diabetic cells.


Encountering extreme weather during migration: individual strategies and their consequences.
The way that animal populations respond to changing weather conditions is a major focus of current research, fuelled by the need to predict the future consequences of climatic changes. Severe weather events can provide valuable opportunities to uncover the mechanisms through which the weather influences population demography but opportunities to track individual responses to such events are rare. Senner et al. (2015) report on an exceptional opportunity to address this issue, when their detailed studies of a migratory shorebird population were interrupted by an extreme weather event that coincided with spring migration, a key period in the annual cycle of migratory species. Through tracking of individuals across the migratory range, Senner et al. (2015) show that, while individual schedules were severely disrupted by the harsh weather, with many individuals undertaking reverse migrations and experiencing delayed breeding, breeding success was unaffected. This study highlights the complexities involved in predicting the ecological consequences of extreme weather events and the key role of behavioural flexibility in mitigating the costs to individuals.


Free Sialic Acid Acts as a Signal That Promotes Streptococcus pneumoniae Invasion of Nasal Tissue and Nonhematogenous Invasion of the Central Nervous System.
Streptococcus pneumoniae (pneumococcus) is a leading cause of bacterial meningitis and neurological sequelae in children worldwide. Acute bacterial meningitis is widely considered to result from bacteremia that leads to blood-brain barrier breakdown and bacterial dissemination throughout the central nervous system (CNS). Previously, we showed that pneumococci can gain access to the CNS through a nonhematogenous route without peripheral blood infection. This access is thought to occur when the pneumococci in the upper sinus follow the olfactory nerves and enter the CNS through the olfactory bulbs. In this study, we determined whether the addition of exogenous sialic acid postcolonization promotes nonhematogenous invasion of the CNS. Previously, others showed that treatment with exogenous sialic acid post-pneumococcal infection increased the numbers of CFU recovered from an intranasal mouse model of infection. Using a pneumococcal colonization model, an in vivo imaging system, and a multiplex assay for cytokine expression, we demonstrated that sialic acid can increase the number of pneumococci recovered from the olfactory bulbs and brains of infected animals. We also show that pneumococci primarily localize to the olfactory bulb, leading to increased expression levels of proinflammatory cytokines and chemokines. These findings provide evidence that sialic acid can enhance the ability of pneumococci to disseminate into the CNS and provide details about the environment needed to establish nonhematogenous pneumococcal meningitis.


The effect of phosphoric acid pre-etching and thermocycling on self-etching adhesive enamel bonding.
When inserting a resin-bonded prosthesis, it is not known whether pumicing, rinsing, and using a self-etching primer (SE) on prepared surfaces is sufficient or whether the use of an additional conventional acid etchant is warranted due to the increased time and risk of contamination. The purpose of this study was to evaluate the effects of pre-etching with phosphoric acid (PA) and thermocycling (TC) on the enamel shear bond strength (SBS) of an autopolymerizing adhesive system on ground enamel. Human teeth were embedded in phenolic rings with methyl methacrylate resin, and their enamel surfaces were ground flat to a 600 grit surface. The teeth were then divided into 4 subsets (n=22): 1) PSN, PA pre-etch with SE, no TC; 2) PST, PA pre-etch with self-etching primer and TC; 3) SEN, self-etching primer alone, no TC; and 4) SET, self-etching primer and TC. A multifactorial study design was used to evaluate 2 factors (pre-etching with PA and TC) at 2 levels (presence or absence) by grouping different subsets. Pre-etch consisted of teeth being etched for 30 seconds with PA, rinsed, re-etched, and rinsed. Self-etching consisted of 60 seconds with ED Primer. Rods of Rexillium III, airborne-particle abraded with 50 μm aluminum oxide, were bonded to enamel surfaces with Panavia 21 OP under a 19.6 N load. Thermocycling consisted of alternating between water baths of 5 ± 2°C and 55 ± 2°C for 5000 cycles. Shear bond strength (SBS) was determined by loading the specimens to failure at a crosshead speed of 1 mm/min. Mean values were analyzed with a 2-way ANOVA (factors were surface treatment and TC) at α=.05. Pre-etching with phosphoric acid showed greater SBS to enamel (P=.028) than the self-etching primer alone. Thermocycling did not have a significant effect (P=.424). There was a significant difference in SBS between pre-etching enamel surfaces with phosphoric acid in addition to the self-etching primer and using the self-etching primer exclusively. Thermocycling did not significantly affect SBS.


Valence shell threshold photoelectron spectroscopy of C3Hx (x = 0-3).
We present the photoelectron spectra of C3Hx (x = 0-3) formed in a microwave discharge flow-tube reactor by consecutive H abstractions from C3H4 (C3Hx + F → C3Hx-1 + HF (x = 1-4)), but also from F + CH4 schemes by secondary reactions. The spectra were obtained combining tunable VUV synchrotron radiation with double imaging electron/ion coincidence techniques, yielding mass-selected threshold photoelectron spectra. The obtained results complement not only existing ones, but for the first time the photoelectron spectra of C3, cyclic and linear C3H (c,l-C3H) as well as of the excited states of C3H3 are reported. In the case of c-C3H, l,t-C3H2 and C3H3, Franck-Condon simulations have been performed in order to assign the vibrational structure. The adiabatic ionization energies of these radicals are reported and compared to ab initio calculated values as well as to theoretical values using known enthalpies of formation.


Membrane insertion of a voltage sensor helix.
Most membrane proteins contain a transmembrane (TM) domain made up of a bundle of lipid-bilayer-spanning α-helices. TM α-helices are generally composed of a core of largely hydrophobic amino acids, with basic and aromatic amino acids at each end of the helix forming interactions with the lipid headgroups and water. In contrast, the S4 helix of ion channel voltage sensor (VS) domains contains four or five basic (largely arginine) side chains along its length and yet adopts a TM orientation as part of an independently stable VS domain. Multiscale molecular dynamics simulations are used to explore how a charged TM S4 α-helix may be stabilized in a lipid bilayer, which is of relevance in the context of mechanisms of translocon-mediated insertion of S4. Free-energy profiles for insertion of the S4 helix into a phospholipid bilayer suggest that it is thermodynamically favorable for S4 to insert from water to the center of the membrane, where the helix adopts a TM orientation. This is consistent with crystal structures of Kv channels, biophysical studies of isolated VS domains in lipid bilayers, and studies of translocon-mediated S4 helix insertion. Decomposition of the free-energy profiles reveals the underlying physical basis for TM stability, whereby the preference of the hydrophobic residues of S4 to enter the bilayer dominates over the free-energy penalty for inserting charged residues, accompanied by local distortion of the bilayer and penetration of waters. We show that the unique combination of charged and hydrophobic residues in S4 allows it to insert stably into the membrane.


Multi-ancestry genome-wide gene-sleep interactions identify novel loci for blood pressure.
Long and short sleep duration are associated with elevated blood pressure (BP), possibly through effects on molecular pathways that influence neuroendocrine and vascular systems. To gain new insights into the genetic basis of sleep-related BP variation, we performed genome-wide gene by short or long sleep duration interaction analyses on four BP traits (systolic BP, diastolic BP, mean arterial pressure, and pulse pressure) across five ancestry groups in two stages using 2 degree of freedom (df) joint test followed by 1df test of interaction effects. Primary multi-ancestry analysis in 62,969 individuals in stage 1 identified three novel gene by sleep interactions that were replicated in an additional 59,296 individuals in stage 2 (stage 1 + 2 Pjoint < 5 × 10-8), including rs7955964 (FIGNL2/ANKRD33) that increases BP among long sleepers, and rs73493041 (SNORA26/C9orf170) and rs10406644 (KCTD15/LSM14A) that increase BP among short sleepers (Pint < 5 × 10-8). Secondary ancestry-specific analysis identified another novel gene by long sleep interaction at rs111887471 (TRPC3/KIAA1109) in individuals of African ancestry (Pint = 2 × 10-6). Combined stage 1 and 2 analyses additionally identified significant gene by long sleep interactions at 10 loci including MKLN1 and RGL3/ELAVL3 previously associated with BP, and significant gene by short sleep interactions at 10 loci including C2orf43 previously associated with BP (Pint < 10-3). 2df test also identified novel loci for BP after modeling sleep that has known functions in sleep-wake regulation, nervous and cardiometabolic systems. This study indicates that sleep and primary mechanisms regulating BP may interact to elevate BP level, suggesting novel insights into sleep-related BP regulation.


Cell signalling - the proteomics of it all.
A challenge for biomedical scientists today is to arrive at an understanding of cellular behavior on a global scale. The advent of DNA microarrays has greatly facilitated discovery of gene expression profiles associated with different cellular states. The problem of understanding cellular signaling at the level of the interacting proteins is in some ways more challenging. Ashman et al. discuss the current methods available for studying protein interactions on a global scale, as well as directions for the future. Technical hurdles exist at many stages, from the isolation of protein complexes, to the determination of their composition, to the software and databases needed to analyze the results of large-scale, high-throughput datasets. Ashman et al. suggest that, with advances in technology and cooperation among academia and industry, a global protein interaction map that underlies cellular behavior will emerge as an essential resource for basic and applied research.


Corosolic acid ameliorates acute inflammation through inhibition of IRAK-1 phosphorylation in macrophages.
Corosolic acid (CA), a triterpenoid compound isolated from Lagerstroemia speciosa L. (Banaba) leaves, exerts anti-inflammatory effects by regulating phosphorylation of interleukin receptor-associated kinase (IRAK)-2 via the NF-κB cascade. However, the protective effect of CA against endotoxic shock has not been reported. LPS (200 ng/mL, 30 min) induced phosphorylation of IRAK-1 and treatment with CA (10 μM) significantly attenuated this effect. In addition, CA also reduced protein levels of NLRP3 and ASC which are the main components of the inflammasome in BMDMs. LPS-induced inflammasome assembly through activation of IRAK-1 was down-regulated by CA challenge. Treatment with Bay11-7082, an inhibitor of IκB-α, had no effect on CA-mediated inhibition of IRAK-1 activation, indicating that CA-mediated attenuation of IRAK-1 phosphorylation was independent of NF-κB signaling. These results demonstrate that CA ameliorates acute inflammation in mouse BMDMs and CA may be useful as a pharmacological agent to prevent acute inflammation. [BMB Reports 2016; 49(5): 276-281].


Microscope spectrometer for light scattering investigations.
We describe a setup including a microscope to study volumes of a few mum(3) by static and dynamic light scattering (DLS) in a backscattering configuration. Light scattered by individual objects of micrometric size can be analyzed in the 400-800?nm spectral range. This setup can also be employed to study both diluted and concentrated colloidal solutions by DLS measurements. For diluted solutions we found evidence of the fluctuations of the number of particles in a confocal volume. We discuss their contribution to the autocorrelation function of the scattered intensity measured as a function of time.


Effectiveness and cost-effectiveness of adding a cognitive behavioral treatment to the rehabilitation of chronic low back pain.
To investigate return to work and cost-effectiveness of the addition of cognitive-behavioral treatment to standard therapy compared to standard 3-week inpatient rehabilitation for patients with chronic low back pain. A prospective economic evaluation alongside a randomized controlled trial was performed. Outcomes included days off work due to spinal complaints, health-related quality of life, and direct and indirect disease-related costs. A total of 409 patients with chronic low back pain, who were admitted to a 3-week inpatient rehabilitation, were randomly assigned to usual care or usual care plus cognitive behavioral treatment. Average incremental costs for psychological treatment during rehabilitation were Euros 127 (95% CI 125.6, 130.9; p < 0.001). Six months after rehabilitation, patients in the intervention group were absent from work an average of 5.4 (95% CI -1.4, 12.1; p = 0.12) days less than patients receiving usual treatment. Between groups, there were no significant differences in quality-adjusted life-years gained or in direct medical or nonmedical costs. The cognitive behavioral treatment showed lower indirect costs: Euros 751 (95% CI -145, 1641; p = 0.097). Adding a cognitive behavioral component to standard therapy may reduce work days lost and thus decrease indirect costs. From a societal perspective, the cost of the psychological treatment was compensated by lower indirect costs.


Neighborhood Deprivation Moderates Shared and Unique Environmental Influences on Hazardous Drinking: Findings from a Cross-Sectional Co-Twin Study.
Background: There has been increased interest in the interplay of genetic and environmental factors in the development of problematic alcohol use, including socioeconomic conditions of the neighborhood. Using a co-twin design, we examined the extent to which contributions of genetic, shared environmental, and unique environmental influences on hazardous drinking differed according to levels of neighborhood socioeconomic deprivation. Method: Data came from 1,521 monozygotic (MZ) and 609 dizygotic (DZ) twin pairs surveyed in Washington State. A measure of neighborhood deprivation was created based on census-tract-level variables and the Alcohol Use Disorders Identification Test 3-item instrument was used to assess level of hazardous drinking. We tested a series of nested structural equation models to examine associations among hazardous drinking, neighborhood deprivation, and the variance components (genetic [A], shared [C] and unique environmental [E] influences) of these two constructs, testing for both main effects and moderation by neighborhood deprivation. Results: Neighborhood deprivation was significantly associated with increased hazardous drinking, after accounting for A and C variance common to both phenotypes. Adjusting for within-pair differences in income and education, neighborhood deprivation moderated the magnitude of variance components of hazardous drinking, with the variance attributable to shared environment and non-shared environment increasing in more deprived neighborhoods. Conclusions: Findings point to amplification of early childhood as well as unique adulthood environmental risk on hazardous drinking in areas of greater deprivation.


Reflection of nanosecond Nd:YAG laser pulses in ablation of metals.
Hemispherical total reflectivity of copper, nickel, and tungsten in ablation by nanosecond Nd:YAG laser pulses in air of atmospheric pressure is experimentally studied as a function of laser fluence in the range of 0.1-100 J/cm(2). Our experiment shows that at laser fluences below the plasma formation threshold the reflectivity of mechanically polished metals remains virtually equal to the table room-temperature reflectivity values. The hemispherical total reflectivity of the studied metals begins to drop at a laser fluence of the plasma formation threshold. With increasing laser fluence above the plasma formation threshold the reflectivity sharply decreases to a low value and then remains unchanged with further increasing laser fluence. Computation of the surface temperature at the plasma formation threshold fluence reveals that its value is substantially below the melting point that indicates an important role of the surface nanostructural defects in the plasma formation on a real sample due to their enhanced heating caused by both plasmonic absorption and plasmonic nanofocusing.


Mechanism and dynamics of translesion DNA synthesis catalyzed by the Escherichia coli Klenow fragment.
Translesion DNA synthesis represents the ability of a DNA polymerase to incorporate and extend beyond damaged DNA. In this report, the mechanism and dynamics by which the Escherichia coli Klenow fragment performs translesion DNA synthesis during the misreplication of an abasic site were investigated using a series of natural and non-natural nucleotides. Like most other high-fidelity DNA polymerases, the Klenow fragment follows the "A-rule" of translesion DNA synthesis by preferentially incorporating dATP opposite the noninstructional lesion. However, several 5-substituted indolyl nucleotides lacking classical hydrogen-bonding groups are incorporated approximately 100-fold more efficiently than the natural nucleotide. In general, analogues that contain large substituent groups in conjunction with significant pi-electron density display the highest catalytic efficiencies ( k cat/ K m) for incorporation. While the measured K m values depend upon the size and pi-electron density of the incoming nucleotide, k cat values are surprisingly independent of both biophysical features. As expected, the efficiency by which these non-natural nucleotides are incorporated opposite templating nucleobases is significantly reduced. This reduction reflects minimal increases in K m values coupled with large decreases in k cat values. The kinetic data obtained with the Klenow fragment are compared to that of the high-fidelity bacteriophage T4 DNA polymerase and reveal distinct differences in the dynamics by which these non-natural nucleotides are incorporated opposite an abasic site. These biophysical differences argue against a unified mechanism of translesion DNA synthesis and suggest that polymerases employ different catalytic strategies during the misreplication of damaged DNA.


Needle knife-assisted endoscopic polypectomy for a large inflammatory fibroid colon polyp by making its stalk into an omega shape using an endoloop.
Colonic inflammatory fibroid polyp (IFP) is an uncommon benign polypoid lesion, which is composed of fibroblasts, numerous small vessels and edematous connective tissue with marked eosinophilic inflammatory cell infiltration. This condition is frequently detected in the stomach and small intestine, but uncommon in the colon. Although IFP is a benign lesion, surgical resections are performed in most colonic cases because the polyps are usually too large to resect endoscopically. Only three patients underwent endoscopic polypectomy in our literature reviews. Here, we present a case of IFP in the descending colon successful endoscopically resected using a novel technique of trapping its stalk with an endoloop, forming the stalk into an omega shape, and then dissecting the stalk with a needle knife.


L-citrulline attenuates arrested alveolar growth and pulmonary hypertension in oxygen-induced lung injury in newborn rats.
Bronchopulmonary dysplasia (BPD) is characterized by arrested alveolar development and complicated by pulmonary hypertension (PH). NO promotes alveolar growth. Inhaled NO (iNO) ameliorates the BPD phenotype in experimental models and in some premature infants. Arginosuccinate synthetase (ASS) and arginosuccinate lyase (ASL) convert L-citrulline to L-arginine; L-citrulline is regenerated during NO synthesis from L-arginine. Plasma levels of these NO precursors are low in PH. We hypothesized that L-citrulline prevents experimental O2-induced BPD in newborn rats. Rat pups were assigned from birth through postnatal day (P) 14 to room air (RA), RA + L-citrulline, 95% hyperoxia (BPD model), and 95%O2 + L-citrulline. Rat pups exposed to hyperoxia had fewer and enlarged air spaces and decreased capillary density, mimicking human BPD. This was associated with decreased plasma L-arginine and L-citrulline concentrations on P7. L-citrulline treatment significantly increased plasma L-arginine and L-citrulline concentrations and increased ASL protein expression in hyperoxia. L-citrulline preserved alveolar and vascular growth in O2-exposed pups and decreased pulmonary arterial medial wall thickness (MWT) and right ventricular hypertrophy (RVH). Increased lung arginase (ARG) activity in O2-exposed pups was reversed by L-citrulline treatment. L-citrulline supplementation prevents hyperoxia-induced lung injury and PH in newborn rats. L-citrulline may represent a novel therapeutic alternative to iNO for prevention of BPD.


Silicon photonic slot waveguide Bragg gratings and resonators.
We present the design, fabrication, and characterization of integrated Bragg gratings in silicon-on-insulator slot waveguides. The Bragg gratings are formed with sidewall corrugations, either on the inside or on the outside of the waveguide. We demonstrate resonators implemented using phase-shifted Bragg gratings in slot waveguides, showing quality factors up to 3 × 10(4). Due to the strong optical confinement in the slot, these devices are promising for optical sensing applications. The devices were fabricated using a CMOS-compatible process, facilitating high-volume and low-cost production.


The evolution of paired appendages in vertebrates: T-box genes in the zebrafish.
The presence of two sets of paired appendages is one of the defining features of jawed vertebrates. We are interested in identifying genetic systems that could have been responsible for the origin of the first set of such appendages, for their subsequent duplication at a different axial level, and/or for the generation of their distinct identities. It has been hypothesized that four genes of the T-box gene family (Tbx2-Tbx5) played important roles in the course of vertebrate limb evolution. To test this idea, we characterized the orthologs of tetrapod limb-expressed T-box genes from a teleost, Danio rerio. Here we report isolation of three of these genes, tbx2, tbx4, and tbx5. We found that their expression patterns are remarkably similar to those of their tetrapod counterparts. In particular, expression of tbx5 and tbx4 is restricted to pectoral and pelvic fin buds, respectively, while tbx2 can be detected at the anterior and posterior margins of the outgrowing fin buds. This, in combination with conserved expression patterns in other tissues, suggests that the last common ancestor of teleosts and tetrapods possessed all four of these limb-expressed T-box genes (Tbx2-Tbx5), and that these genes had already acquired, and have subsequently maintained, their gene-specific functions. Furthermore, this evidence provides molecular support for the notion that teleost pectoral and pelvic fins and tetrapod fore- and hindlimbs, respectively, are homologous structures, as suggested by comparative morphological analyses.


Shared and unique microbes between Small hive beetles (Aethina tumida) and their honey bee hosts.
The small hive beetle (SHB) is an opportunistic parasite that feeds on bee larvae, honey, and pollen. While SHBs can also feed on fruit and other plant products, like its plant-feeding relatives, SHBs prefer to feed on hive resources and only reproduce inside bee colonies. As parasites, SHBs are inevitably exposed to bee-associated microbes, either directly from the bees or from the hive environment. These microbes have unknown impacts on beetles, nor is it known how extensively beetles transfer microbes among their bee hosts. To identify sets of beetle microbes and the transmission of microbes from bees to beetles, a metagenomic analysis was performed. We identified sets of herbivore-associated bacteria, as well as typical bee symbiotic bacteria for pollen digestion, in SHB larvae and adults. Deformed wing virus was highly abundant in beetles, which colonize SHBs as suggested by a controlled feeding trial. Our data suggest SHBs are vectors for pathogen transmission among bees and between colonies. The dispersal of host pathogens by social parasites via floral resources and the hive environment increases the threats of these parasites to honey bees.


Normal FDG distribution patterns in the head and neck: PET/CT evaluation.
To retrospectively evaluate the distribution of fluorine 18 fluorodeoxyglucose (FDG) in the head and neck region with combined positron emission tomography-computed tomography (PET/CT) in patients with no known abnormality in this region. The institutional review board allowed a retrospective review of PET/CT images obtained in 78 patients with non-head and neck cancer and waived the requirement for informed consent. The accumulation of FDG in 11 normal head and neck structures was visually and quantitatively assessed retrospectively. Positive rate percentage (PRP) was defined as the sum of the percentages of patients with grade 2 and grade 3 tracer uptake intensity. Standardized uptake values (SUVs) were calculated for quantitative analysis. Mean SUVs were compared between the male and female patients by using the unpaired t test, and the correlation between FDG uptake and patient age was assessed by using the Pearson correlation coefficient test. Intense tracer uptake was usually seen in the palatine tonsils (PRP, 98%; mean SUV, 3.48), soft palate (PRP, 96%; mean SUV, 3.13), and lingual tonsils (PRP, 96%; mean SUV, 3.11). In the inferior concha (PRP, 4%; mean SUV, 1.56), thyroid gland (PRP, 3%; mean SUV, 1.31), and tongue (PRP, 1%; mean SUV, 1.39), uptake was typically minimal. FDG accumulation was variable in the sublingual glands (PRP, 72%; mean SUV, 2.93), spinal cord (PRP, 64%; mean SUV, 2.12), submandibular glands (PRP, 53%; mean SUV, 2.11), parotid glands (PRP, 51%; mean SUV, 1.90), and vocal cords (PRP, 19%; mean SUV, 1.77). The mean normal-tissue SUV in the soft palate was higher in male than in female patients (P <.01). A negative correlation between age and physiologic FDG uptake was seen in the palatine tonsils (r=-0.51, P <.001) and sublingual glands (r=-0.70, P <.001). Intense FDG uptake was usually observed in the palatine tonsils, lingual tonsils, and soft palate, whereas uptake in the major salivary glands was variable.


Miniaturisation and simplification of solid-state proton activity sensors for non-aqueous media and ionic liquids.
This work is the further development of the previous pH (effective) sensor work where a biologically derived proton-active redox centre - riboflavin (RFN) - was entrapped into a vapour phase polymerised poly(3,4-ethylenedioxythiophene) film and ferrocene (Fc) dissolved in the sample solution was used as an internal reference redox couple. Here, we report a disposable solid state pH (effective) sensor where we successfully incorporated both RFN and Fc into a single solid state electrode. The electrodes were then used for pH (effective) sensing where water is not required. The system was further miniaturised and simplified from a 3 electrode to a 2 electrode setup with the working electrode area being as small as 0.09 mm(2). The sensors show the ability to measure pH (effective) in both aqueous and non-aqueous media including ionic liquids (ILs) regardless of their hydrophobicity. This is an important step towards the ability to customise ILs or non-aqueous media with suitable proton activity (PA) for various applications e.g. customised ILs for biotechnological applications such as protein preservation and customised media for PA dependent reactions such as catalytic reactions.


Complex Gaussian representation of statistical pulses.
We develop a general representation for ensembles of non-stationary random pulses in terms of statistically uncorrelated, time-delayed, frequency-shifted Gaussian pulses which are classical counterparts of coherent states of a quantum harmonic oscillator. We show that the two-time correlation function describing second-order statistics of the pulses can be expanded in terms of the complex Gaussian pulses. We also demonstrate how the novel formalism can be applied to describe recently introduced Gaussian Schell-model pulses and pulse trains generated by typical mode-locked lasers.


Randomised evaluation of government health programmes does present a challenge to standard research ethics frameworks.
In a recent issue of Journal of Medical Ethics (JME), we discussed the ethical review of evaluations of interventions that would occur whether or not the evaluation was taking place. We concluded that standard research ethics frameworks including the Ottawa Statement, which requires justification for all aspects of an intervention and its roll-out, were a poor guide in this area. We proposed that a consideration of researcher responsibility, based on the consequences of the research taking place, would be a more appropriate way delineate the scope of research ethics review. Weijer and Taljaard present a counterargument to our proposal, which we address in this reply. They claim that a focus on researcher responsibility will weaken the protection of research participants and link it to 'unethical research' and a 'government experimenting on its own people'. However, the moral responsibility of researchers is defined in terms of the consequences of the research on human welfare and harm, not in opposition to it. Weijer and Taljaard argue that researchers must justify what they are studying whether or not they have any control over it and that governments must justify their programmes, including by demonstrating equipoise, to a research ethics committee if they implement them in a randomised way. We strongly disagree that this is a defensible way to define the scope of research ethics review and argue that this provides no further protections to research participants beyond what we propose, but places a potential barrier to learning from government programmes.


A Web-Based Platform for Designing Vaccines against Existing and Emerging Strains of Mycobacterium tuberculosis.
Development of an effective vaccine against drug-resistant Mycobacterium tuberculosis (Mtb) is crucial for saving millions of premature deaths every year due to tuberculosis. This paper describes a web portal developed for assisting researchers in designing vaccines against emerging Mtb strains using traditional and modern approaches. Firstly, we annotated 59 genomes of Mycobacterium species to understand similarity/dissimilarity between tuberculoid, non-tuberculoid and vaccine strains at genome level. Secondly, antigen-based vaccine candidates have been predicted in each Mtb strain. Thirdly, epitopes-based vaccine candidates were predicted/discovered in above antigen-based vaccine candidates that can stimulate all arms of immune system. Finally, a database of predicted vaccine candidates at epitopes as well at antigen level has been developed for above strains. In order to design vaccine against a newly sequenced genome of Mtb strain, server integrates three modules for identification of strain-, antigen-, epitope-specific vaccine candidates. We observed that 103,522 unique peptides (9mers) had the potential to induce an antibody response and/or promiscuous binder to MHC alleles and/or have the capability to stimulate T lymphocytes. In summary, this web-portal will be useful for researchers working on designing vaccines against Mtb including drug-resistant strains. The database is available freely at http://crdd.osdd.net/raghava/mtbveb/.


Radiocontrast medium-induced declines in renal function: a role for oxygen free radicals.
Intrarenal injection of radiocontrast medium (RCM) results in transient vasoconstriction and a persistent decline in glomerular filtration rate (GFR). Adenosine modulates this vasoconstrictor response and is postulated to increase oxygen free radical (OFR) generation. We hypothesized that the persistent decline in (GFR that follows RCM administration results in an increased generation of OFR. We evaluated the effects of RCM injection on renal blood flow, inulin clearance, hypoxanthine, xanthine, and malondialdehyde concentrations in four groups of non-volume-expanded, pentobarbital sodium anesthetized dogs in the presence and absence of intravenous allopurinol, 25 mg/min (group 1), intrarenal superoxide dismutase (SOD), 400 U/min (group 2), heat-inactivated intrarenal SOD, 400 U/min (group 3), and simultaneous infusions of intrarenal SOD, 400 U/min, to one kidney and saline to the other (group 4). Both allopurinol and SOD significantly attenuated the fall in GFR after RCM administration over control. Malondialdehyde concentrations were attenuated over control in all treated groups, indicating a decrease in OFR generation. We conclude that intrarenal injection of RCM results in increased production of OFR. Inhibition of OFR production by allopurinol and increased OFR removal by SOD attenuates the effects of RCM on declines in GFR.


Collagen-Coated Bovine Bone in Peri-implantitis Defects: A Pilot Study on a Novel Approach.
As dental implants have become routine therapy, clinicians are more frequently being faced with treating peri-implantitis. To date, no single treatment protocol has been shown to be the preferred means to treat peri-implantitis. The aim of this retrospective case series is to present a novel approach utilizing porcine collagen-coated bovine bone (CBB) to treat peri-implantitis. Eleven patients, with no history of periodontitis, presenting with peri-implantitis around a single restored dental implant, were included in the study. At initial and follow-up examinations, bleeding on probing (BOP), probing depth (PD), and gingival margin location (GM) were recorded. Following surgical debridement of the peri-implant defect and treatment of the implant surface with a 0.12% chlorhexidine gluconate solution, bony defects were grafted with CBB. All patients had 12 months of follow-up. Upon presentation, average PD at the deepest site (DS) was 7.6 ± 1.9 mm. At the time of surgery, excess cement was found around nine implants (81%). All patients healed uneventfully without postoperative complications. At 6 and 12 months, all implants showed favorable results with average DS PD reduction of 3.9 ± 1.5 mm and 4.1 ± 1.6 mm, respectively. All implants showed radiographic signs of bone fill, while GM showed no changes from preoperative measurements at either 6 (0.1 ± 0.5 mm) or 12 (0.0 ± 0.6 mm) months. The use of a porcine collagen-coated bovine bone graft to treat peri-implantitis represents a potentially predictable therapeutic modality. Randomized controlled trials are necessary to substantiate the treatment outcomes.


Carboxylating enzymes and pathway of photosynthetic carbon assmilation in different marine algae-Evidence for the C4-pathway?
Experiments on short-term photosynthesis in H(14)CO3 (-) (2-5 s) using various species of different algal classes resulted in predominant (14)C-labelling (>90% of total (14)C-incorporation) of phosphorylated compounds. The percentage of malate and aspartate usually accounts for distinctly less than 10% of the total (14)C-labelling. These findings are consistent with data from enzymatic analyses, since 97-100% of the carboxylation capacity is due to ribulose-1.5-biphosphate carboxylase (EC 22.214.171.124) in Rhodophyceae and Chlorophyceae. Phaeophyceae are generally characterized by considerable activity of phosphoenolpyruvate carboxykinase (EC 188.8.131.52): at least 10% of carboxylation is confined to this enzyme. Similar ratios are obtained when rates of photosynthesis and of light-independent CO2-fixation are compared. Activity of phosphoenolpyruvate carboxylase (EC 22.214.171.124) could not be detected in the species investigated. The results are discussed with emphasis on the pathway of photosynthetic carbon assimilation in marine algae.


The tobacco industry's thwarting of marketing restrictions and health warnings in Lebanon.
This article outlines how the tobacco industry has undermined tobacco control efforts in Lebanon since the early 1970s. An analysis of online and on-site tobacco industry documents, reviews of newspapers, policy and other documents, and interviews with key policy makers were conducted. Findings reveal how the weakness of tobacco control legislation in Lebanon has been the product of an effective tobacco industry strategy to weaken the content and scope of regulation, and delay adoption and implementation. The tobacco industry has built and maintained strong alliances that were and are regularly mobilised to effectively oppose regulation. Despite ratification of the World Health Organization Framework Convention on Tobacco Control in 2005, Lebanon's tobacco control track remains weak. Public health professionals and the government should work hard to oppose such tobacco industry tactics.