Isolation of Acanthamoeba isolates belonging to T2, T3, T4 and T7 genotypes from environmental samples in Ankara, Turkey

Acanthamoeba keratitis is a blinding infection that is becoming increasingly important in human health. Early diagnosis is a prerequisite for successful treatment and requires identification of Acanthamoeba at the genotypic level. The genus Acanthamoeba consists of both pathogenic and non-pathogenic species and has been recently classified into 13 different genotypes, T1-T12 and T14. More importantly, 95% of Acanthamoeba isolates that produce keratitis belong to T4 genotypes. In this study, we attempted to determine whether predominance of T4 isolates in Acanthamoeba keratitis is due to greater virulence or greater prevalence. We isolated 18 Acanthamoeba isolates from environmental samples in Ankara, Turkey and determined their pathogenic potential by means osmotolerance, temperature tolerance and in vitro cytotoxicity assays using corneal epithelial cells. Ribosomal DNA sequencing revealed that 10 isolates belong to T2, 5 belong to T3, 2 belong to T4 and one belongs to T7 genotype. As expected, T3 and T4 isolates exhibited the most pathogenic traits and were osmotolerant, temperature tolerant and exhibited severe corneal epithelial cell cytotoxicity indicating their pathogenic potential. Overall these data indicate that high frequency of T4 isolates in keratitis cases may well be due to their greater virulence. This is the first report presenting environmental distribution of Acanthamoeba in Ankara, Turkey

Drugs for discoid lupus erythematosus

BACKGROUND: Discoid lupus erythematosus (DLE) is a chronic form of cutaneous lupus, which can cause scarring. Many drugs have been used to treat this disease and some (such as thalidomide, cyclophosphamide and azathioprine) are potentially toxic. This is an update of a Cochrane Review first published in 2000, and previously updated in 2009. We wanted to update the review to assess whether any new information was available to treat DLE, as we were still unsure of the effectiveness of available drugs and how to select the most appropriate treatment for an individual with DLE. OBJECTIVES: To assess the effects of drugs for discoid lupus erythematosus. SEARCH METHODS: We updated our searches of the following databases to 22 September 2016: the Cochrane Skin Specialised Register, CENTRAL, MEDLINE, Embase and LILACS. We also searched five trials databases, and checked the reference lists of included studies for further references to relevant trials. Index Medicus (1956 to 1966) was handsearched and we approached authors for information about unpublished trials. SELECTION CRITERIA: We included all randomised controlled trials (RCTs) of drugs to treat people with DLE in any population group and of either gender. Comparisons included any drug used for DLE against either another drug or against placebo cream. We excluded laser treatment, surgery, phototherapy, other forms of physical therapy, and photoprotection as we did not consider them drug treatments. DATA COLLECTION AND ANALYSIS: At least two reviewers independently extracted data onto a data extraction sheet, resolving disagreements by discussion. We used standard methods to assess risk of bias, as expected by Cochrane. MAIN RESULTS: Five trials involving 197 participants were included. Three new trials were included in this update. None of the five trials were of high quality.'Risk of bias' assessments identified potential sources of bias in each study. One study used an inappropriate randomisation method, and incomplete outcome data were a concern in another as 15 people did not complete the trial. We found most of the trials to be at low risk in terms of blinding, but three of the five did not describe allocation concealment.The included trials inadequately addressed the primary outcome measures of this review (percentage with complete resolution of skin lesions, percentage with clearing of erythema in at least 50% of lesions, and improvement in patient satisfaction/quality of life measures).One study of fluocinonide cream 0.05% (potent steroid) compared with hydrocortisone cream 1% (low-potency steroid) in 78 people reported complete resolution of skin lesions in 27% (10/37) of participants in the fluocinonide cream group and in 10% (4/41) in the hydrocortisone group, giving a 17% absolute benefit in favour of fluocinonide (risk ratio (RR) 2.77, 95% CI 0.95 to 8.08, 1 study, n = 78, low-quality evidence). The other primary outcome measures were not reported. Adverse events did not require discontinuation of the drug. Skin irritation occurred in three people using hydrocortisone, and one person developed acne. Burning occurred in two people using fluocinonide (moderate-quality evidence).A comparative trial of two oral agents, acitretin (50 mg daily) and hydroxychloroquine (400 mg daily), reported two of the outcomes of interest: complete resolution was seen in 13 of 28 participants (46%) on acitretin and 15 of 30 participants (50%) on hydoxychloroquine (RR 0.93, 95% CI 0.54 to 1.59, 1 study, n = 58, low-quality evidence). Clearing of erythema in at least 50% of lesions was reported in 10 of 24 participants (42%) on acitretin and 17 of 25 (68%) on hydroxychloroquine (RR 0.61, 95% CI 0.36 to 1.06, 1 study, n = 49, low-quality evidence). This comparison did not assess improvement in patient satisfaction/quality of life measures. Participants taking acitretin showed a small increase in serum triglyceride, not sufficient to require withdrawal of the drug. The main adverse effects were dry lips (93% of the acitretin group and 20% of the hydroxychloroquine group) and gastrointestinal disturbance (11% of the acitretin group and 17% of the hydroxychloroquine group). Four participants on acitretin withdrew due to gastrointestinal events or dry lips (moderate-quality evidence).One trial randomised 10 people with DLE to apply a calcineurin inhibitor, pimecrolimus 1% cream, or a potent steroid, betamethasone 17-valerate 0.1% cream, for eight weeks. The study reported none of the primary outcome measures, nor did it present data on adverse events.A trial of calcineurin inhibitors compared tacrolimus cream 0.1% with placebo (vehicle) over 12 weeks in 14 people, but reported none of our primary outcome measures. In the tacrolimus group, five participants complained of slight burning and itching, and for one participant, a herpes simplex infection was reactivated (moderate-quality evidence).Topical R-salbutamol 0.5% cream was compared with placebo (vehicle) over eight weeks in one trial of 37 people with DLE. There was a significant improvement in pain and itch in the salbutamol group at two, four, six, and eight weeks compared to placebo, but the trial did not record a formal measure of quality of life. None of the primary outcome measures were reported. Changes in erythema did not show benefit of salbutamol over placebo, but we could not obtain from the trial report the number of participants with clearing of erythema in at least 50% of lesions. There were 15 events in the placebo group (experienced by 12 participants) and 24 in the salbutamol group (experienced by nine participants). None of the adverse events were considered serious (moderate-quality evidence). AUTHORS' CONCLUSIONS: Fluocinonide cream may be more effective than hydrocortisone in clearing DLE skin lesions. Hydroxychloroquine and acitretin appear to be of equal efficacy in terms of complete resolution, although adverse effects might be more frequent with acitretin, and clearing of erythema in at least 50% of lesions occurred less often in participants applying acitretin. Moderate-quality evidence found adverse events were minor on the whole. There is not enough reliable evidence about other drugs used to treat DLE. Overall, the quality of the trials and levels of uncertainty were such that there is a need for further trials of sufficient duration comparing, in particular, topical steroids with other agents

Evidence-based sizing of non-inferiority trials using decision models

Abstract Background There are significant challenges to the successful conduct of non-inferiority trials because they require large numbers to demonstrate that an alternative intervention is “not too much worse” than the standard. In this paper, we present a novel strategy for designing non-inferiority trials using an approach for determining the appropriate non-inferiority margin (δ), which explicitly balances the benefits of interventions in the two arms of the study (e.g. lower recurrence rate or better survival) with the burden of interventions (e.g. toxicity, pain), and early and late-term morbidity. Methods We use a decision analytic approach to simulate a trial using a fixed value for the trial outcome of interest (e.g. cancer incidence or recurrence) under the standard intervention (pS) and systematically varying the incidence of the outcome in the alternative intervention (pA). The non-inferiority margin, pA – pS = δ, is reached when the lower event rate of the standard therapy counterbalances the higher event rate but improved morbidity burden of the alternative. We consider the appropriate non-inferiority margin as the tipping point at which the quality-adjusted life-years saved in the two arms are equal. Results Using the European Polyp Surveillance non-inferiority trial as an example, our decision analytic approach suggests an appropriate non-inferiority margin, defined here as the difference between the two study arms in the 10-year risk of being diagnosed with colorectal cancer, of 0.42% rather than the 0.50% used to design the trial. The size of the non-inferiority margin was smaller for higher assumed burden of colonoscopies. Conclusions The example demonstrates that applying our proposed method appears feasible in real-world settings and offers the benefits of more explicit and rigorous quantification of the various considerations relevant for determining a non-inferiority margin and associated trial sample size.https://deepblue.lib.umich.edu/bitstream/2027.42/146777/1/12874_2018_Article_643.pd

Determination of the scale of pattern and distribution in Helicteres isora L (Sterculiaceae)

Helicteres isora L. is a traditional multipurpose plant used by indigenous community and villagers inall the three major climatic zones in Sri Lanka. It naturally occurs in the edges of forests and indisturbed secondary vegetation. It is fastly disappearing in the wet zone due to land clearing and highextraction rates. The present study was conducted to understand the pattern and the scale of distributionof H. isora in order to provide information for biodiversity conservation and further to enable thesustainable use.Twelve natural populations were identified in wet, intermediate and dry zones and the distribution ofindividuals was studied using gradient directed transect method. The t test was performed for eachpopulation to detect the pattern of distribution and pattern analysis was carried out to determine thescale of pattern.Out of twelve populations surveyed, only five populations showed contagious distribution (p < 0.05)while seven populations showed random pattern of distribution. This indicates that the populations ofH isora do not fall into a particular pattern of distribution in nature. This may be due to the highdisturbance present in and around the populations.The results of the pattern analysis reveal more peaks in smaller block sizes (2m2) and larger blocksizes (32m2) indicating aggregated pattern in respective block sizes. Peaks in smaller block sizes aredue to the morphology of the plant as it produces new plants from roots. Peaks at larger block sizesare due to the extrinsic factors and these results could be utilized in the in- situ conservation of Hisora .

Frequency of natural out-crossing in partially cleistogamous pigeonpea lines in diverse environments

Natural out-crossing is the major cause of loss of varietal purity in pegeonpea [Cajanus cajan (L.) Millsp.]. The frequency of natural out-crossing of partially cleistogamous mutant lines, characterized by a modified keel and filamentous anthers, was studied at two locations in Sri Lanka and three locations in India. Indeterminate growth habit and normal floral morphology were used as dominant markers and the frequency of anural out-crossing was estimated as percentage of the observed hybrid plants. Natural out-crossing in the mutant lines in Sri Lanka ranged from 0.14 to 1.33% in comparison to 6.34 to 19.64% in the controls. In the Indian environments, natural outcrossing ranged from 0.16 to 2.67%. The mutant was higly stable over diverse environments, and may be of considerable economic importance in pigeonpea improvement and seed-production programs

A multiorganism pipeline for antiseizure drug discovery:Identification of chlorothymol as a novel γ-aminobutyric acidergic anticonvulsant

OBJECTIVE:Current medicines are ineffective in approximately one-third of people with epilepsy. Therefore, new antiseizure drugs are urgently needed to address this problem of pharmacoresistance. However, traditional rodent seizure and epilepsy models are poorly suited to high-throughput compound screening. Furthermore, testing in a single species increases the chance that therapeutic compounds act on molecular targets that may not be conserved in humans. To address these issues, we developed a pipeline approach using four different organisms. METHODS:We sequentially employed compound library screening in the zebrafish, Danio rerio, chemical genetics in the worm, Caenorhabditis elegans, electrophysiological analysis in mouse and human brain slices, and preclinical validation in mouse seizure models to identify novel antiseizure drugs and their molecular mechanism of action. RESULTS:Initially, a library of 1690 compounds was screened in an acute pentylenetetrazol seizure model using D rerio. From this screen, the compound chlorothymol was identified as an effective anticonvulsant not only in fish, but also in worms. A subsequent genetic screen in C elegans revealed the molecular target of chlorothymol to be LGC-37, a worm γ-aminobutyric acid type A (GABAA ) receptor subunit. This GABAergic effect was confirmed using in vitro brain slice preparations from both mice and humans, as chlorothymol was shown to enhance tonic and phasic inhibition and this action was reversed by the GABAA receptor antagonist, bicuculline. Finally, chlorothymol exhibited in vivo anticonvulsant efficacy in several mouse seizure assays, including the 6-Hz 44-mA model of pharmacoresistant seizures. SIGNIFICANCE:These findings establish a multiorganism approach that can identify compounds with evolutionarily conserved molecular targets and translational potential, and so may be useful in drug discovery for epilepsy and possibly other conditions

Primary coronary artery bypass surgery in the presence of decreasing preoperative renal function: effect on short-term outcomes

Background: This study evaluated the impact of decreasing renal function on short-term outcomes in patients undergoing primary coronary artery bypass grafting (CABG). Methods: The study period was from February 1999 to February 2009. Data on 4050 patients undergoing primary CABG were prospectively collected and analyzed retrospectively. The study population was divided into 3 groups: the CABG:N group, patients with preoperative serum creatinine levels 2 mg/dL (n = 87); and the CABG:D group, patients on dialysis (n = 16). Results: The significant differences between the groups (CABG:D > CABG:RF > CABG:N) in short-term outcomes were with respect to blood product use (P < .001), postoperative acute myocardial infarction (P < .001), pulmonary complications (P .001), infection (P < .001), and death (P < .001). The risk of short-term death (30 days) in the CABG:D group (4/16, 25%) was 25 times greater than that in the CABG:N group (38/3947, 0.96%). Conclusion: CABG in the presence of renal failure is associated with significant morbidity and mortality

Evaluation of pigeonpea accessions and selected lines for reaction to Maruca

Maruca vitrata (Geyer) is a serious insect pest of tropical legumes. In Sri Lanka, yield losses due to Maruca damage in pigeonpea [Cajanus cajan (L.) Millsp.] range up to 100%. The development of resistant cultivars and germplasm is one of the best means of control. The objectives of this study were to screen 271 accessions for resistance to M. vitrata and evaluate reaction of lines selected from the promising accessions. The high level of natural incidence of Maruca in Sri Lanka provided an opportunity for evaluation of germplasm at Field Crops Research and Development Institute, Maha Illuppallama. Screening of the germplasm accessions revealed large variation in Maruca damage to flowers and pods. On average, the Maruca damage in determinate accessions (66–75%) was higher than that of nondeterminate accessions (41–50%). Resistant plants from four determinate and 12 nondeterminate accessions were selected. Further selection for resistance to Maruca damage among and within lines derived from the resistant plants was exercised for six generations under nonsprayed field conditions. Under insecticide-free conditions, the selections from two accessions showed significant yield advantages over controls. Data on pod damage and larval counts indicated that the resistance was conditioned through yield compensation mechanisms. In pigeonpea, this is the first report of the selection of Maruca resistant lines. Further studies showed that by using the resistant genotypes it is possible to reduce the number of insecticide sprays for economic yields

The pre-concept design of the DEMO tritium, matter injection and vacuum systems

In the Pre-Concept Design Phase of EU-DEMO, the work package TFV (Tritium – Matter Injection – Vacuum) has developed a tritium self-sufficient three-loop fuel cycle architecture. Driven by the need to reduce the tritium inventory in the systems to an absolute minimum, this requires the continual recirculation of gases in loops without storage, avoiding hold-ups of tritium in each process stage by giving preference to continuous over batch technologies, and immediate use of tritium extracted from tritium breeding blankets. In order to achieve this goal, a number of novel concepts and technologies had to be found and their principal feasibility to be shown. This paper starts from a functional analysis of the fuel cycle and introduces the results of a technology survey and ranking exercise which provided the prime technology candidates for all system blocks. The main boundary conditions for the TFV systems are described based on which the fuel cycle architecture was developed and the required operational windows of all subsystems were defined. To validate this, various R&D lines were established, selected results of which are reported, together with the key technology developments. Finally, an outlook towards the Concept Design Phase is given