Profiling cytotoxic microRNAs in pediatric and adult glioblastoma cells by high-content screening, identification, and validation of miR-1300.

MicroRNAs play an important role in the regulation of mRNA translation and have therapeutic potential in cancer and other diseases. To profile the landscape of microRNAs with significant cytotoxicity in the context of glioblastoma (GBM), we performed a high-throughput screen in adult and pediatric GBM cells using a synthetic oligonucleotide library representing all known human microRNAs. Bioinformatics analysis was used to refine this list and the top seven microRNAs were validated in a larger panel of GBM cells using state-of-the-art in vitro assays. The cytotoxic effect of our most relevant candidate was assessed in a preclinical model. Our screen identified ~100 significantly cytotoxic microRNAs with 70% concordance between cell lines. MicroRNA-1300 (miR-1300) was the most potent and robust candidate. We observed a striking binucleated phenotype in miR-1300 transfected cells due to cytokinesis failure followed by apoptosis. This was also observed in two stem-like patient-derived cultures. We identified the physiological role of miR-1300 as a regulator of endomitosis in megakaryocyte differentiation where blockade of cytokinesis is an essential step. In GBM cells, where miR-1300 is normally not expressed, the oncogene Epithelial Cell Transforming 2 (ECT2) was validated as a direct key target. ECT2 siRNA phenocopied the effects of miR-1300, and ECT2 overexpression led to rescue of miR-1300 induced binucleation. We showed that ectopic expression of miR-1300 led to decreased tumor growth in an orthotopic GBM model. Our screen provides a resource for the neuro-oncology community and identified miR-1300 as a novel regulator of endomitosis with translatable potential for therapeutic application.

Diagnosis of copy number variation by Illumina next generation sequencing is comparable in performance to oligonucleotide array comparative genomic hybridisation.

Array comparative genomic hybridisation (aCGH) profiling is currently the gold standard for genetic diagnosis of copy number. Next generation sequencing technologies provide an alternative and adaptable method of detecting copy number by comparing the number of sequence reads in non-overlapping windows between patient and control samples. Detection of copy number using the BlueGnome 8×60k oligonucleotide aCGH platform was compared with low resolution next generation sequencing using the Illumina GAIIx on 39 patients with developmental delay and/or learning difficulties who were referred to the Leeds Clinical Cytogenetics Laboratory. Sensitivity and workflow of the two platforms were compared. Customised copy number algorithms assessed sequence counts and detected changes in copy number. Imbalances detected on both platforms were compared. Of the thirty-nine patients analysed, all eleven imbalances detected by array CGH and confirmed by FISH or Q-PCR were also detected by CNV-seq. In addition, CNV-seq reported one purported pathogenic copy number variant that was not detected by array CGH. Non-pathogenic, unconfirmed copy number calls were detected by both platforms; however few were concordant between the two. CNV-seq offers an alternative to array CGH for copy number analysis with resolution and future costs comparable to conventional array CGH platforms and with less stringent sample requirements.

The use of local anaesthesia in haemorrhoidal banding: a randomized controlled trial.

AIM: Rubber band ligation is a common office procedure for the treatment of symptomatic haemorrhoids. It can be associated with pain and vasovagal symptoms. The effect of local anaesthetic use during banding was studied. METHOD: A single-blinded randomized controlled trial was carried out in the colorectal outpatient clinic. Patients presenting with symptomatic haemorrhoids suitable for banding were prospectively recruited and randomized to undergo the procedure with local anaesthetic or without (control). Submucosal bupivacaine was injected immediately after banding just proximal to the site. Vasovagal symptoms were assessed at the time of banding and pain scores (visual analogue scale) were recorded at the conclusion of the procedure, after 15 min, and on leaving the clinic. RESULTS: Seventy-two patients (40 local anaesthetic injection, group 1; 32 no injection, group 2) were recruited. The mean ages were 50 and 54 years respectively, the median duration of symptoms was 12 months in each group and the median number of haemorrhoids banded was three in each group. The mean pain score on leaving the clinic was 2.6 (95% CI 2.1, 3.1) in group 1 and 4.1 (95% CI 3.3, 5.0) (P = 0.04) in group 2. There were no complications related to local anaesthetic use. No significant difference in vasovagal symptoms was found (P = 0.832). CONCLUSION: Local anaesthetic injection at the time of banding is simple and safe. It may reduce patient discomfort following banding of haemorrhoids.

Surgical versus medical methods for second trimester induced abortion.

BACKGROUND: Determining the optimal method of performing second-trimester abortions is important, since they account for a disproportionate amount of abortion-related morbidity and mortality. OBJECTIVES: To compare surgical and medical methods of inducing abortion in the second trimester of pregnancy with regard to efficacy, side effects, adverse events, and acceptability. SEARCH STRATEGY: We identified trials using Pub Med, EMBASE, POPLINE, and the Cochrane Central Register of Controlled Trials (CENTRAL). We also searched the reference lists of identified studies, relevant review articles, book chapters, and conference proceedings for additional, previously unidentified studies. We contacted experts in the field for information on other published or unpublished research. SELECTION CRITERIA: Randomised trials comparing any surgical to any medical method of inducing abortion at >/= 13 weeks' gestation were included. DATA COLLECTION AND ANALYSIS: We assessed the validity of each study using the methods suggested in the Cochrane Handbook. Investigators were contacted as needed to provide additional information regarding trial conduct or outcomes. Two reviewers abstracted the data. Odds ratios and 95% confidence intervals were calculated for dichotomous variables using RevMan 4.2. The trials did not have uniform interventions, therefore, we were unable to combine them into a meta-analysis. MAIN RESULTS: Two studies met criteria for this review. One compared dilation and evacuation (D&E) to intra-amniotic instillation of prostaglandin F(2) (alpha). The second study compared D&E to induction with mifepristone and misoprostol. Compared with prostaglandin instillation, the combined incidence of minor complications was lower with D&E (OR 0.17, 95% CI 0.04-0.65) as was the total number of minor and major complications (OR 0.12, 95% CI 0.03-0.46). The number of women experiencing adverse events was also lower with D&E than with mifepristone and misoprostol (OR 0.06, 95% CI 0.01-0.76). Although women treated with mifepristone and misoprostol reported significantly more pain than those undergoing D&E, efficacy and acceptability were the same in both groups. In both trials, fewer subjects randomised to D&E required overnight hospitalisation. AUTHORS' CONCLUSIONS: Dilation and evacuation is superior to instillation of prostaglandin F(2) (alpha). The current evidence also appears to favour D&E over mifepristone and misoprostol, however larger randomised trials are needed.

Candesartan cilexetil: comparison of once-daily versus twice-daily administration for systemic hypertension. Candesartan Cilexetil Study Investigators.

This randomized, double-masked, placebo-controlled, forced-titration, parallel-arm study was designed to compare the blood pressure (BP)-lowering effect of candesartan cilexetil, a potent antagonist of the angiotensin II receptor subtype AT1, administered once daily with that of the same agent administered twice daily at the same total daily dose of 16 mg. After a 4- to 5-week placebo run-in period, 277 patients with a sitting diastolic BP of 95 to 109 mm Hg were randomly allocated to receive placebo (n = 92) or candesartan cilexetil 8 mg once daily for 4 weeks, followed by forced titration to either 16 mg once daily (n = 91) or 8 mg twice daily (n = 94) for 4 weeks. At 8 weeks, mean reductions in trough sitting diastolic BP were similar for the once- and twice-daily treatment groups (9.4 and 10.3 mm Hg, respectively). After 8 weeks of treatment, no statistically significant differences were observed in diastolic or systolic BP, peak or trough BP, or sitting or standing BP between the 2 active-treatment groups. The rates of positive responses (defined as a trough sitting diastolic BP of <90 mm Hg or a decrease in BP of > or =10 mm Hg) were also similar (approximately 60%) in the once- and twice-daily candesartan cilexetil groups. Furthermore, placebo-corrected trough-to-peak ratios for sitting diastolic BP exceeded 75% for both candesartan cilexetil regimens, indicating a persistent 24-hour duration of drug effect. Ambulatory BP monitoring performed in a subset of patients (n = 44) confirmed the consistent 24-hour BP-lowering effect and preservation of diurnal variation with once-daily dosing. No significant between-group differences were observed in the incidence or severity of clinical or laboratory adverse events. The results of this study suggest that identical daily doses of candesartan cilexetil administered once or twice daily have comparable efficacy and tolerability and that no additional clinical benefit is derived from twice-daily administration.

Genetic relatedness of North American populations of Tomicus piniperda (Coleoptera:Scolytidae).

We used DNA fingerprinting by random amplified polymorphic (RAPD) DNA and electrophoretic characterization of esterase isozymes to investigate the genetic relatedness of North American populations of the exotic bark beetle Tomicus piniperda (L.). Cluster analyses of genetic distances among populations identified the Illinois population as an outlier population with mean genetic distances to other populations averaging 0.895 (where complete dissimilarity = 2), compared with genetic distance averages of 0.595 among populations excluding Illinois. When genetic distance means and geographical distance between populations were compared, the results suggested that T. piniperda populations in the United States were established separately in Illinois near Lake Michigan and in Ohio along Lake Erie. Molecular markers indicated that insects derived from the 2 founder groups were interbreeding in contiguous regions in western Indiana.

Encapsulated cell technology.

The potential therapeutic applications of encapsulated cells are enormous. In the US alone, it has been estimated that nearly half-a-trillion dollars are spent each year to care for patients who suffer tissue loss or dysfunction. Over 6 million patients suffer from neurodegenerative disorders such as Alzheimer's disease and Parkinson's disease, over 14 million patients suffer from diabetes, and millions more from liver failure, hemophilia, and other diseases caused by the loss of specific vital cellular functions. It appears likely that by the end of the decade clinical trials of encapsulated cells to treat many of these diseases will become a reality. The Food and Drug Administration has already authorized studies to evaluate the safety and biological activity of several types of systems. A number of issues will have to be addressed, including the sourcing of raw materials, the design and building of manufacturing facilities, the scale-up and optimization process, storage and distribution of the product, and quality control.

Potentially hazardous co-prescribing of beta-adrenoceptor antagonists and agonists in the community.

AIM: The aim of this study was to investigate the co-prescribing of beta-antagonists and beta-agonists in the community, and to assess the potential hazards of such co-prescribing. METHODS: The study was set in the population of Tayside, Scotland (population approximately 400,000), between January 1993 and March 1993. An automated person-specific prescribing database was used, which could also be linked to hospital admissions. Patients who were co-prescribed beta-antagonists and beta-agonists on the same day or within 30 days were selected. A model was used to identify those who showed an asthmatic profile, on the basis of age, and previous prescribing and hospitalization history, and for whom the co-prescribing was judged to be particularly hazardous. RESULTS: Altogether, 0.9% of 15824 patients who received a beta-antagonist during the study period received a beta-agonist on the same day. This figure increased to 274 (1.7%) for 30-day co-prescription. A few instances of particularly hazardous co-prescribing were identified, which involved young people who had previously received prescriptions for corticosteroids and been hospitalized for asthma. CONCLUSION: Potentially hazardous co-prescribing of beta-agonists and beta-antagonists occurs despite labelled warnings, even in patients who appear to be at high risk. These events are quite rare but probably should not occur at all.

Repellent properties of the host compound 4-allylanisole to the southern pine beetle.

The phenylpropanoid 4-allylanisole is a compound produced by loblolly pines (Pinus taeda L.), an abundant species in southern pine forests and a preferred host of southern pine beetle (Dendroctonus frontalis Zimmermann). Repellency of individual beetles was demonstrated in laboratory behavioral assays ofD. frontalis and other scolytids. Inhibition was demonstrated in natural populations ofD. frontalis using baited traps. In both tests, response to the inhibitory pheromone verbenone was used for comparison. In the laboratory, a higher proportion of newly emerged and reemergedD. frontalis responded negatively to 4-allylanisole than to verbenone. However, fewer reemergent than newly emerged individuals responded to either compound. In all field trials, the response ofD. frontalis to its attractant pheromone in funnel traps was significantly reduced by simultaneous release of 4-allylanisole. In most trials total reduction did not differ from verbenone; however, unlike verbenone, 4-allylanisole reduced male and female catches proportionally. Both compounds together did not significantly further reduce trap catch. The response of a major predator,Thanasimus dubius (F.), to the attractant pheromone ofD. frontalis, did not differ with the simultaneous release of either verbenone or 4-allylanisole. The results of preliminary field applications are presented and discussed.

Factors associated with failure to complete treatment for diabetic retinopathy among Navajo Indians.

OBJECTIVE: To determine the proportion of patients with suspected proliferative diabetic retinopathy who did not receive the recommended follow-up ophthalmological evaluation and care, and to examine associations between various patient characteristics and the failure to obtain care. RESEARCH DESIGN AND METHODS: The study cohort included all Navajo Indians identified by a retrospective review of records who had proliferative diabetic retinopathy diagnosed at an Indian Health Service Optometry Clinic between 1 October 1985 and 30 September 1988. Follow-up data were obtained by medical record reviews and by interviews with subjects. RESULTS: Of 69 patients identified, 57 of 61 living patients were interviewed. Twenty-three (40.4%) had failed to obtain recommended follow-up. The RR for incomplete treatment among those without a vehicle in the household compared with those with a vehicle was 1.91 (95% CI 1.32-2.76). Other factors associated with incomplete treatment were female sex and marital status other than currently married. Twelve (21%) patients answered "no" to the question, "Have you been told that diabetes was affecting your eyes?" Eight of 38 (21%) who confirmed that they had been told that diabetes was affecting their eyes responded "no" to the question, "Do you think that diabetes is affecting your eyes?" However, the answers to these questions did not distinguish between patients who obtained or did not obtain recommended care. CONCLUSIONS: Interventions to increase the proportion of Navajo Indians with diabetic retinopathy who receive appropriate ophthalmologic care must address the issue of transportation.

Quantitative genetic tools for insecticide resistance risk assessment: estimating the heritability of resistance.

Quantitative genetic studies of resistance can provide estimates of genetic parameters not available with other types of genetic analyses. Three methods are discussed for estimating the amount of additive genetic variation in resistance to individual insecticides and subsequent estimation of the heritability (h2) of resistance. Sibling analysis and offspring-parent regression permit direct estimates of h2 by comparing the resistance phenotypes of individuals of known relatedness. Threshold trait analyses, performed on data from selection experiments, provide estimates of realized heritability. Procedures are outlined for predicting changes in resistance to insecticides based on h2 estimates. Quantitative genetic theory is examined as it relates to resistance and resistance as a quantitative trait; quantitative genetic methods also are unique in providing estimates of genetic correlations between traits. Comments are included on estimates of genetic correlation between resistance and phenotypic traits (e.g., development time) and how they may be used to predict changes in the genetic aspects of phenology that result from insecticide applications (i.e., to predict how the reproductive capacity of future generations will differ from that of the treated generation).