Interference microscopy delineates cellular proliferations on flat mounted internal limiting membrane specimens.

Aim: To demonstrate that interference microscopy of flat mounted internal limiting membrane specimens clearly delineates cellular proliferations at the vitreomacular interface. Methods: ILM specimens harvested during vitrectomy were fixed in glutaraldehyde 0.05% and paraformaldehyde 2% for 24 h (pH 7.4). In addition to interference microscopy, immunocytochemistry using antibodies against glial fibrillar acidic protein (GFAP) and neurofilament (NF) was performed. After washing in phosphatebuffered saline 0.1 M, the specimens were flat-mounted on glass slides without sectioning, embedding or any other technique of conventional light microscopy. A cover slide and 49,6-diamidino-2-phenylindole (DAPI) medium were added to stain the cell nuclei. Results: Interference microscopy clearly delineates cellular proliferations at the ILM. DAPI stained the cell nuclei. Areas of cellular proliferation can be easily distinguished from ILM areas without cells. Immunocytochemistry can be performed without changing the protocols used in conventional microscopy. Conclusion: Interference microscopy of flat mounted ILM specimens gives new insights into the distribution of cellular proliferations at the vitreomacular interface and allows for determination of the cell density at the ILM. Given that the entire ILM peeled is seen en face, the techniques described offer a more reliable method to investigate the vitreoretinal interface in terms of cellular distribution compared with conventional microscopy

Cholesterol and Malondialdehyde Contents of Broiler-Chicken Meat Supplemented with Indigofera Zolingeriana Top Leaf Meal

This research aimed to increase functional value of broiler-chicken meat containing high antioxidant and low cholesterol through substitution of soybean meal (SBM) with Indigofera zollingeriana top leaf meal (ILM). The experiment used 160 day old broiler chicken (Cobb strain). The experimental chicken were provided dietary treatments when they were 15-days old (initial body weight of 460.5±1.56 g/bird) and terminated on day 35. A completely randomized design (CRD) with four treatments and four replications and ten birds in each replication was used in this experiment. Dietary treatments were: R1= diet containing 20% soybean meal (SBM) without I. zollingeriana top leaf meal (ILM); R2= diet containing 16% SBM and 5.9% ILM; R3= diet containing 12% SBM and 11.8% ILM; R4= diet containing 8% SBM and 17.74% ILM. Variables measured were performances (feed consumption, body weight gain, and feed conversion) and the quality of broiler meat (cholesterol, fat content, and malondialdehyde [MDA] concentration). The results showed that supplementation of 17.74% ILM (R4) as the substitution of 60% soybean meal protein produced the same performances of broilers as those of control diet (R1). Supplementation of 11.8% ILM as the substitution of 40% soybean meal protein (R3) decreased meat cholesterol by 34.70%, meat fat content by 52.93%, and MDA concentration by 62.52%. The conclusion of this study was that supplementation of 17.74% ILM produced the same performances as that of control diet, increased antioxidant content of the meat, indicated by a lower MDA concentration, and decreased cholesterol, as well as fat content of broiler-chicken meat

Sequential epiretinal membrane removal with internal limiting membrane peeling in brilliant blue G-assisted macular surgery

Purpose To assess the selectivity of brilliant blue G (BBG) staining by analysing the morphological components of unstained and stained tissue obtained during epiretinal membrane (ERM) removal with internal limiting membrane (ILM) peeling in BBG-assisted macular surgery. Methods Twenty-six surgical specimens were removed from 13 eyes with epiretinal gliosis during vitrectomy using BBG for ERM and ILM peeling. We included eyes with idiopathic macular pucker, idiopathic macular hole and vitreomacular traction syndrome. The dye was injected into the fluid-filled globe. Unstained and stained epiretinal tissue was harvested consecutively and placed into separate containers. All specimens were processed for conventional transmission electron microscopy. Results The first surgical specimen of all eyes showed no intraoperative staining with BBG and corresponded to masses of cells and collagen. The second surgical specimen demonstrated good staining characteristics and corresponded to the ILM in all patients included. In seven eyes, the ILM specimens were seen with minor cell proliferations such as single cells or a monolayer of cells. Myofibroblasts, fibroblasts and astrocytes were present. In five cases, native vitreous collagen fibrils were found at the ILM. In six of the eyes, ILM specimens were blank. Conclusion Our clinicopathological correlation underlines the selective staining properties of BBG. The residual ILM is selectively stained by BBG even when a small amount of cells and collagen adheres to its vitreal side. To reduce the retinal exposure to the dye, the surgeon might choose to remove the ERM without using the dye, followed by a BBG injection to identify residual ILM

Staining and peeling of the internal limiting membrane using a fluorescent dye (Rhodamine 6 G)

Aim: To assess whether low concentrations of a fluorescent dye such as Rhodamine 6G would help the unaided human eye visualise the vitreous and the internal limiting membrane (ILM) under standard halogen illumination.Material/methods: The UV/Vis absorption (E) and fluorescence (I) spectra of Rhodamine 6G in water were measured and compared with Indocyanine Green (ICG). Surgery was performed in two rhesus monkeys and consisted of standard pars plana vitrectomy with halogen light source used for illumination. Rhodamine 6G was diluted in balanced salt solution (BSS). A few drops of the dye in a concentration of 0.1% (307 mOsm) were applied over the posterior pole in the air-filled globe and washed out by irrigation after 1 min. Immediately after surgery, the globes were enucleated, fixated and prepared for histological evaluation.Results: In contrast to ICG, both the maximum of the absorption and emission of Rhodamin 6G are very much within the spectral sensitivity of the human eye. The Rhodamine 6G--BSS itself appears red in colour. Using a dye concentration of 0.1%, there was no visible red-staining of the ILM as such. As the dye was irrigated out with BSS, a marked green fluorescence of the fluid within the vitreous cavity was noted. With halogen illumination through a standard 20-gauge light pipe, the dye provided a sufficient green fluorescence to identify and safely remove the ILM and to clearly differentiate areas of peeled from non-peeled ILM. During light microscopy, eyes revealed a peeled ILM demarcation with no signs of acute retinal toxicity.Conclusion: The findings indicate that a fluorescent dye can be used for ILM peeling. Assuming that the fluorophore provides a high enough fluorescence quantum yield after adsorption to the ILM, much lower dye concentrations could be used compared with absorbent dyes, thereby minimising toxic effects

Towards an understanding of third-order galaxy-galaxy lensing

Third-order galaxy-galaxy lensing (G3L) is a next generation galaxy-galaxy lensing technique that either measures the excess shear about lens pairs or the excess shear-shear correlations about lenses. It is clear that these statistics assess the three-point correlations between galaxy positions and projected matter density. For future applications of these novel statistics, we aim at a more intuitive understanding of G3L to isolate the main features that possibly can be measured. We construct a toy model ("isolated lens model"; ILM) for the distribution of galaxies and associated matter to determine the measured quantities of the two G3L correlation functions and traditional galaxy-galaxy lensing (GGL) in a simplified context. The ILM presumes single lens galaxies to be embedded inside arbitrary matter haloes that, however, are statistically independent ("isolated") from any other halo or lens position. In the ILM, the average mass-to-galaxy number ratio of clusters of any size cannot change. GGL and galaxy clustering alone cannot distinguish an ILM from any more complex scenario. The lens-lens-shear correlator in combination with second-order statistics enables us to detect deviations from a ILM, though. This can be quantified by a difference signal defined in the paper. We demonstrate with the ILM that this correlator picks up the excess matter distribution about galaxy pairs inside clusters. The lens-shear-shear correlator is sensitive to variations among matter haloes. In principle, it could be devised to constrain the ellipticities of haloes, without the need for luminous tracers, or maybe even random halo substructure. [Abridged]Comment: 14 pages, 3 figures, 1 table, accepted by A&A; some "lens-shear-shear" were falsely "lens-lens-shear

Comparison of Three Different Techniques of Inverted Internal Limiting Membrane Flap in Treatment of Large Idiopathic Full-Thickness Macular Hole.

Purpose: To evaluate and compare three different techniques of inverted internal limiting membrane (ILM) flap in the treatment of large idiopathic full-thickness macular hole. Methods: In a comparative interventional case series, 72 eyes from 72 patients with large (\u3e 400 µm) full-thickness macular hole were randomly enrolled into three different groups: group A - hemicircular ILM peel with temporally hinged inverted flap; group B - circular ILM peel with temporally hinged inverted flap; and group C - circular ILM peel with superior inverted flap. Best-corrected visual acuity (BCVA), anatomical closure rate, and ellipsoid zone (EZ) or external limiting membrane (ELM) defects were evaluated preoperatively, at week 1, and months 1, 3 and 6 after surgery. Results: There were 24 eyes in group A, 23 in group B, and 25 in group C. In all three groups, larger diameter macular hole was associated with worse preoperative visual acuity (r=0.625, P\u3c0.001). Mean BCVA improved significantly in all three groups 6 months after surgery (0.91vs 0.55, p\u3c0.001). 6 months after surgery, mean BCVA improved from 0.91 logMAR to 0.52±0.06 in group A, 0.90 to 0.53±0.06 in group B, and 0.91 to 0.55±0.11 in group C. In group A vs. B vs. C, improvement of BCVA was 0.380±0.04 vs. 0.383±0.04 vs. 0.368±0.11 logMAR, with no statistically significant difference between groups (P=0.660). The rate of successful hole closure was 87.5% vs. 91.3% vs. 100%. Although the closure rate was 100% in Group C (circular ILM peel with superiorly hinged inverted flap), this difference was not statistically significant (P=0.115). Conclusion: ILM peel with an inverted flap is a highly effective procedure for the treatment of large, full-thickness macular hole. Different flap techniques have comparable results, indicating that the technique can be chosen based on surgeon preference

Information and display requirements for independent landing monitors

The ways an Independent Landing Monitor (ILM) may be used to complement the automatic landing function were studied. In particular, a systematic procedure was devised to establish the information and display requirements of an ILM during the landing phase of the flight. Functionally, the ILM system is designed to aid the crew in assessing whether the total system (e.g., avionics, aircraft, ground navigation aids, external disturbances) performance is acceptable, and, in case of anomaly, to provide adequate information to the crew to select the least unsafe of the available alternatives. Economically, this concept raises the possibility of reducing the primary autoland system redundancy and associated equipment and maintenance costs. The required level of safety for the overall system would in these cases be maintained by upgrading the backup manual system capability via the ILM. A safety budget analysis was used to establish the reliability requirements for the ILM. These requirements were used as constraints in devising the fault detection scheme. Covariance propagation methods were used with a linearized system model to establish the time required to correct manually perturbed states due to the fault. Time-to-detect and time-to-correct requirements were combined to devise appropriate altitudes and strategies for fault recovery