Chemical enhancers of posttranscriptional gene silencing in Arabidopsis.

RNAi mediated by small-interfering RNAs (siRNAs) operates via transcriptional (TGS) and posttranscriptional gene silencing (PTGS). In Arabidopsis thaliana, TGS relies on DICER-LIKE-3 (DCL3)-dependent 24-nt siRNAs loaded into AGO4-clade ARGONAUTE effector proteins. PTGS operates via DCL4-dependent 21-nt siRNAs loaded into AGO1-clade proteins. We set up and validated a medium-throughput, semi-automatized procedure enabling chemical screening, in a 96-well in vitro format, of Arabidopsis transgenic seedlings expressing an inverted-repeat construct from the phloem companion cells. The ensuing quantitative PTGS phenotype was exploited to identify molecules, which, upon topical application, either inhibit or enhance siRNA biogenesis/activities. The vast majority of identified modifiers were enhancers, among which Sortin1, Isoxazolone, and [5-(3,4-dichlorophenyl)furan-2-yl]-piperidine-1-ylmethanethione (DFPM) provided the most robust and consistent results, including upon their application onto soil-grown plants in which their effect was nonautonomous and long lasting. The three molecules increased the RNAi potency of the inverted-repeat construct, in large part by enhancing 21-nt siRNA accumulation and loading into AGO1, and concomitantly reducing AGO4 and DCL3 levels in planta. A similar, albeit not identical effect, was observed on 22-nt siRNAs produced from a naturally occurring inverted-repeat locus, demonstrating that the molecules also enhance endogenous PTGS. In standardized assays conducted in seedling extracts, the three enhancers selectively increased DCL4-mediated processing of in vitro-synthesized double-stranded RNAs, indicating the targeting of a hitherto unknown PTGS component probably independent of the DCL4-cofactor DOUBLE-STRANDED RNA-BINDING 4 (DRB4). This study establishes the proof-of-concept that RNAi efficacy can be modulated by chemicals in a whole organism. Their potential applications and the associated future research are discussed.

Adjustable surgical treatment of adult exotropia: postoperative target angles and surgical success.

OBJECTIVES: This study had 3 objectives: (i) to characterize clinical profiles of adults with consecutive exotropia (CXT), intermittent exotropia (IXT), and sensory exotropia (SXT); (ii) to correlate immediate postoperative target angles with successful long-term ocular alignment; and (iii) to compare the efficacy of adjustable versus nonadjustable medial rectus resection ± advancement. STUDY DESIGN: Retrospective, observational, and interventional cohort study. PARTICIPANTS: A total of 133 adult exotropic patients treated surgically at 3 different hospitals between July 2012 and June 2013. METHODS: The patients were divided according to clinical profiles (CXT, IXT, and SXT) based on ophthalmic and orthoptic assessments. Two treatment groups were established: group I-adjustable medial rectus resection ± advancement and adjustable lateral rectus recession; group II-nonadjustable medial resection ± advancement and adjustable lateral rectus recession. Measurements of immediate postadjustment alignment (target angle) and 4-6 months of follow-up alignment were performed and compared between groups. Surgical success was defined as distance primary position alignment within 10 prism diopters (PD) of orthotropia 4-6 months postoperatively. RESULTS: Comparison of clinical profile groups showed that CXT patients had more hyperopia and amblyopia and smaller preoperative deviations; IXT patients had more diplopia and larger preoperative deviations (near > distance); and SXT patients had poor vision in the deviating eye and larger preoperative deviations. Immediate postoperative alignment was 5.2 PD of esodeviation in group I and 3.2 PD of esodeviation in group II. Overall success rates for ocular alignment at 4-6 months postoperatively were comparable with both surgical techniques (74.6% for group I and 74.3% for group II). Patients with a preoperative deviation ≥40 PD had a lower surgical success rate (63.8%) than patients with a deviation <40 PD (80%). Patients presenting with a significant (-1 or worse) abduction deficit in the operated eye at their first visit after surgery had a better success rate at 4-6 months' follow-up (83.3% vs 67.8%). CONCLUSIONS: Adjustable and nonadjustable medial rectus surgeries seem equally successful. Creation of an abduction deficit in the early postoperative period seems predictive of a better outcome. Larger preoperative angles (≥40 PD) were associated with more exotropic drift and a lower percentage of surgical success. Future studies will continue to search for surgical strategies and the ideal target angle that will produce the best long-term alignment stability.

Retinal function assessment of trypan blue versus indocyanine green assisted internal limiting membrane peeling during macular hole surgery.

OBJECTIVE: To compare multifocal electroretinography (mfERG) retinal function and the anatomical and visual outcomes of macular hole surgery performed with indocyanine green (ICG) or trypan blue (TB). DESIGN: Prospective, randomized study. PARTICIPANTS: Twenty-five eyes of 24 patients. METHODS: Patients underwent a pars plana vitrectomy with removal of the internal limiting membrane. In 14 eyes, internal limiting membrane visualization during macular hole repair was performed using TB, and ICG was used in 11 eyes. The examination protocol (performed before surgery and at 3 weeks, 3 months, 6 months, and 12 months after surgery) included optical coherence tomography, mfERG (mfERG-103 hexagons), and assessment of best corrected visual acuity (BCVA) and contrast sensitivity (CS). RESULTS: Closure of macular hole was achieved in 100% of the cases. In the TB group, P1 amplitude and implicit time improved significantly at 12 months after surgery (P < 0.05), whereas in the ICG group, significant improvement occurred at both 6 (P < 0.05) and 12 months (P < 0.01). BCVA improved significantly in both groups at 6 and 12 months (P < 0.01). Both groups also showed a statistically significant CS improvement at spatial frequency of 6 cycles per degree (P = 0.01) 1 year postoperatively. At 12 months, improvement of P1 amplitude and implicit time, BCVA, and CS was not different between groups. CONCLUSIONS: In this study, the use of TB or ICG appears to yield similar improvement in terms of BCVA, CS, and mfERG amplitude and implicit time changes at 12 months.

Distribution of silver nanoparticles in pregnant mice and developing embryos.

The objective of this study was to evaluate the distribution of silver nanoparticles (NPs) in pregnant mice and their developing embryos. Silver NPs (average diameter 50 nm) were intravenously injected into pregnant CD-1 mice on gestation days (GDs) 7, 8, and 9 at dose levels of 0, 35, or 66 µg Ag/mouse. Mice were euthanised on GD10, and tissue samples were collected and analysed for silver content. Compared with control animals injected with citrate buffer vehicle, silver content was significantly increased (p < 0.05) in nearly all tissues from silver NP-treated mice. Silver accumulation was significantly higher in liver, spleen, lung, tail (injection site), visceral yolk sac, and endometrium compared with other organs from silver NP-treated mice. Furthermore, silver NPs were identified in vesicles in endodermal cells of the visceral yolk sac. In summary, the results demonstrated that silver NPs distributed to most maternal organs, extra-embryonic tissues, and embryos, but did not accumulate significantly in embryos.

Acquired restrictive strabismus and high axial myopia: diagnosis and management.

OBJECTIVE: This paper documents the clinical, ultrasound, magnetic resonance imaging (MRI), and anatomic characteristics of 6 patients with esohypotropia and high axial myopia. The results of strabismus surgery performed on these patients are evaluated. STUDY DESIGN: Retrospective cohort study. PARTICIPANTS: Six patients with esohypotropia and high axial myopia were selected from the practice of the second author (Michael Flanders). METHODS: We extracted the history and data from the preoperative and postoperative ophthalmic and orthoptic examinations, recorded the surgical procedures, and tabulated the postoperative results. A- and B-scans, orbital MRI images, and photo documentation were obtained for all patients. Supramaximal recession and resection on the horizontal rectus muscles, using adjustable sutures, was performed in all patients. A surgical outcome was deemed successful if, in primary position, postoperative horizontal ocular alignment measured 15 prism diopters (PD) or less and (or) if diplopia was eliminated. RESULTS: Echographic and MRI findings demonstrated an enlarged globe in all cases. Globe flattening on the orbital walls was found in 5 cases. Lateral rectus depression was visualized on MRI images in 4 cases. Surgical success was obtained in all patients. The average preoperative primary position esotropia measured 60 PD (25 to 90 PD) and the esodeviation after surgery was 7.0 PD (0 to 15 PD). Ductions were also improved. CONCLUSIONS: Different surgical approaches to realignment of eyes with high myopia and esohypotropia have been attempted. In this study, supramaximal amounts of recession and resection on the rectus muscles provided satisfactory results.