bluuebunny/crossref_metadata_2025 huggingface dataset:
Biorefinery for combined production of jet fuel and ethanol from lipid‐producing sugarcane: a techno‐economic evaluation
Authors: Kumar Deepak , Long Stephen P. , Singh Vijay
Published: September 2017

Replacing fossil fuels with an economically viable green alternative at scale has proved most challenging in the aviation sector. Recently sugarcane, the most productive crop on the planet, has been engineered to accumulate lipids. This opens the way for production of far more industrial vegetable oil per acre than previously possible. This study performs techno‐economic feasibility analysis of jet fuel production from this new cost efficient and high yield feedstock. A comprehensive process model for biorefinery producing hydrotreated jet fuel (from lipids) and ethanol (from sugars), with 1 600 000 MT yr−1lipid‐cane processing capacity, was developed inSuperProDesigner. Considering lipid‐cane development is continuing for higher oil concentrations, analysis was performed with lipid‐cane containing 5%, 10%, 15%, and 20% lipids. Capital investments for the biorefinery ranged from 238.1 to 351.2 millionUSD, with jet fuel capacities of 12.6–50.5 million liters (correspondingly ethanol production of nil to 102.6 million liters). The production cost of jet fuel for different scenarios was estimated $0.73 to $1.79 per liter ($2.74 to $6.76 per gal) of jet fuel. In all cases, the cost of raw materials accounted for more than 70% of total operational cost. Biorefinery was observed self‐sustainable for steam and electricity requirement, because of in‐house steam and electricity generation from burning of bagasse. Minimum fuel selling prices with a 10% discount rate for 20% lipid case was estimated $1.40/L ($5.31/gal), which was lower than most of the reported prices of renewable jet fuel produced from other oil crops and algae. Along with lower production costs, lipid‐cane could produce as high as 16 times the jet fuel (6307 L ha−1) per unit land than that of other oil crops and do so using low‐value land unsuited to most other crops, while being highly water and nitrogen use efficient.
Comprehensive histopathological comparison of epidermotropic/dermal metastatic melanoma and primary nodular melanoma
Authors: Skala Stephanie L , Arps David P , Zhao Lili , Cha Kelly B , Wang Min , Harms Paul W , Andea Aleodor A , Fullen Douglas R , Chan May P
Published: September 2017

Aims

Metastatic melanoma involving the epidermis and/or upper dermis may show significant histological overlap with primary cutaneous melanoma, especially the nodular subtype. Proper histopathological classification is crucial to appropriate staging and management, but is often challenging. The aim of this study was to identify helpful histopathological features for differentiating epidermotropic/dermal metastatic melanoma (EDMM) and primary nodular melanoma (PNM).

Methods and results

A cohort of EDMMs (n = 74) and PNMs (n = 75) was retrospectively reviewed for various histopathological features, and the data were compared between groups by the use of univariate analysis. Features significantly associated with EDMM included a tumour size of <2 mm, an absence of tumour‐infiltrating lymphocytes and plasma cells, monomorphism, and involvement of adnexal epithelium. Features associated with PNM included a polypoid (exophytic) configuration, prominent tumour‐infiltrating plasma cells (TIPs), a tumour size of >10 mm, ulceration, epidermal collarettes, a higher mitotic rate, necrosis, multiple phenotypes, significant pleomorphism, and lichenoid inflammation. In multivariate analysis, a logistic regression model including large tumour size, ulceration, prominent TIPs, lichenoid inflammation and epidermal collarettes was highly predictive of PNM. Six (8%) EDMMs from three patients showed an ‘epidermal‐only’ or ‘epidermal‐predominant’ pattern closely simulating in‐situ or microinvasive melanoma. Two of these cases were tested by fluorescence in‐situ hybridisation, which confirmed clonal relationships with their corresponding primary melanomas.

Conclusions

This is the first comprehensive histopathological comparison of EDMM and PNM. Recognition of the above histopathological associations should aid in the correct classification and staging of cutaneous melanoma. Epidermotropic metastatic melanomas may occasionally show an epidermal‐only/epidermal‐predominant pattern; accurate diagnosis requires prudent clinical correlation and, when necessary, ancillary molecular tests.
Diagnostic accuracy of immunofluorescence versus immunoperoxidase staining to distinguish immune complex‐mediated glomerulonephritis and C3 dominant glomerulopathy
Authors: Bouatou Yassine , Kers Jesper , Chevalier‐Florquin Marie S N , Claessen Nike , Nguyen Tri Q , Damman Jeffrey , Proença Henrique , Roelofs Joris J T H , Florquin Sandrine
Published: September 2017

Aims

The technique used for classification of membranoproliferative glomerulonephritis (MPGN) has been changed from an electron microscopy‐based to an immunofluorescence (IF)‐based semiquantitative technique with immunoperoxidase (IP) staining as a backup option when IF is not possible. Since data on that matter is lacking, our aims were to study the interobserver variability, the correlation and the reclassification of MPGN based on these two techniques.

Methods and results

We retrospectively analysed cases of type 1 MPGN. We repeated IF staining and performed IP staining for IgG, kappa, lambda, C3c and C4d in 35 renal biopsies, among which 19 biopsies had matched IP and IF samples. We observed substantial to near‐perfect agreement among the seven observers for both IF and IP (W coefficients from 0.66 for IF lambda to 0.89 for IF C4d). Of the 19 cases with matched IP and IF samples, five (26%) turned out to have different diagnoses on IF and on IP. Also, the ability of C4d to discriminate immune complex‐mediated glomerulonephritis (ICGN) from C3 glomerulopathy (C3G) was poor, with areas under the curve of 0.44 [95% confidence interval (CI) 0.24–0.63] and 0.66 (95% CI 0.50–0.81) for the receiver operating characteristic curves of IF and IP respectively. Limitations include the fact that no clinical data regarding complement activation were available.

Conclusion

The diagnosis of ICGN versus C3GN depends on the immunochemical technique used. Also, the use of C4d failed to discriminate ICGN from C3G in our study. Further validation studies are required to avoid misdiagnosis based on kidney biopsy.
Properties of BioRoot RCS, a tricalcium silicate endodontic sealer modified with povidone and polycarboxylate
Authors: Siboni F. , Taddei P. , Zamparini F. , Prati C. , Gandolfi M. G.
Published: September 2017

Aim

To evaluate the chemical and physical properties of a tricalcium silicate root canal sealer containing povidone and polycarboxylate (BioRoot RCS), a calcium silicate MTA‐based sealer containing a salicylate resin (MTA Fillapex), a traditional eugenol‐containing sealer (Pulp Canal Sealer) and an epoxy resin‐based root canal sealer (AH Plus).

Methodology

Calcium release, pH, setting time, water sorption, volume of open pores, volume of impervious portion, apparent porosity and weight loss were measured. The ability to nucleate calcium phosphates (CaP) after ageing 28 days in a simulated body fluid was evaluated using ESEM‐EDX and micro‐Raman spectroscopy. Data were statistically analysed (P = 0.05) using one‐way anova (setting time, radiopacity, solubility, water sorption, porosity) or two‐way anova (ion release tests).

Results

BioRoot RCS had a final setting time of 300 min and adequate radiopacity (5.2 mm Al). It demonstrated the highest (P < 0.05) and more prolonged ability to release calcium ions (721 ppm at 3 h) and to increase the pH (11–12) (P < 0.05); B‐type carbonated apatite deposits were found on aged BioRoot RCS (biointeractivity‐related CaP‐forming ability). A final setting time of 270 min and good calcium release (17.4 ppm at 3 h) were measured for MTA Fillapex; apatite deposits were present on aged samples. No calcium release and no alkalizing activity were measured for Pulp Canal Sealer and AH Plus; no CaP nucleation was detected on aged Pulp Canal Sealer, and some apatite deposits were found on aged AH Plus (chemi/physisorption‐related CaP‐deposition). Higher and significantly different (P < 0.05) porosity, water sorption and solubility were measured for the two calcium silicate sealers, especially for BioRoot RCS.

Conclusions

BioRoot RCS had bioactivity with calcium release, strong alkalizing activity and apatite‐forming ability, and adequate radiopacity.
Internal limiting membrane peeling in retinal detachment complicated by grade B proliferative vitreoretinopathy
Authors: Foveau P. , Leroy B. , Ameloot F. , Berrod J.P.
Published: September 2017

Purpose

To specify the benefits of internal limiting membrane (ILM) peeling as a surgical adjunct in primary repair of retinal detachments (RD) complicated by grade B proliferative vitreoretinopathy (PVR).

Methods

A retrospective comparative study included consecutive patients who underwent vitrectomy for primary RD complicated by grade B PVR between May 2010 and December 2015 at Nancy University Hospital (France). All patients were treated with SF6 or C2F6 gaz tamponade. The ILM was routinely peeled after staining from 2012. The best‐corrected visual acuity and spectral‐domain optical coherence tomography (SD‐OCT) were collected at 1 and 3 months postoperatively, looking for epiretinal membrane formation, macular oedema or photoreceptor damage.

Results

Thirty seven eyes who underwent ILM peeling (group 1) and 38 eyes without ILM peeling (group 2) were included. At the end of follow‐up, anatomic success after single surgery was higher in group 1 (89%) than in group 2 (66%, p=0.03). Mean final visual acuity was 0.41 ±0.40 logMAR in group 1 versus 0.43 ±0.22 logMAR in group 2 (p=0.82). After 3 months follow up, we found no epiretinal membrane (ERM) formation on OCT scans in group 1 whereas 5 ERM (20%) were detected in group 2 (p=0.012). The 2 groups did not differ in terms of cystoid macular oedema occurence, macular thickness or photoreceptor damage.

Conclusions

ILM peeling at the macula during vitrectomy for the treatment of retinal detachment complicated by grade B PVR may prevent a second surgery for redetachment or macular pucker.
Subject specific angular waist of the optic nerve head nerve fiber layer allows follow up detection of local nerve fiber bundle loss
Authors: Soderberg P. , Malmberg F. , Sandberg‐Melin C.
Published: September 2017

Purpose

To clarify if frontal plane angular Pigment epithelium central limit‐ Inner limit of the retina Minimal Distance (PIMD) measured with OCT has enough specific angular structure for detection of change of specific angular segments of PIMD over time within glaucoma subjects.

Methods

A Topcon OCT‐2000 was used for capture of the 3D representations of the ONH. from one eye of each of totally 13 subjects with early stage primary open angle glaucoma. Altogether, 3 volumes were captured at two occasions within 3 months. PIMD was semi‐automatically segmented in each volume in 500 equally spaced angular segments distributed over 2Pi in the frontal plane.

Results

Plots of the angular distribution of PIMD for all subject revealed specific low angular frequency overlaid with higher intermediate angular frequencies specific for each subject. High angular frequencies appeared similar for all subjects. A fast Fourier transform of the angular PIMD revealed a high to low frequency content ratio, 0.12, consistent for all segmentations of PIMD‐2Pi. The rotational displacment of angular PIMD in the frontal plane among segmentations, volumes and occasions was determined with cross‐correlation. Then, all PIMD‐2Pi were phase adjusted over segments, volumes and occasions. Finally, the rotation of PIMD‐2Pi was adjusted to a standard rotation. This allows comparison of angular segments of PIMD‐2Pi between visits. The magnitude of different sources of variation were estimated with analysis of variance.

Conclusions

Angular correction in the frontal plane of recordings of angular PIMD of the ONH allows comparison of angular segments of PIMD‐2Pi. This substantially increases the resolution of of an angular segment of PIMD. The resulting precision suggest that detection of an angular segment of PIMD is associated with better resolution than Humphrey visual field MD.
Pathophysiology of uveitis
Authors: Dick A.
Published: September 2017

This talk will overview the pathophysiology of non‐infectious uveitis in relation to recent SUN (standardised uveitis nomenclature) disease classification.The experimental and translational human evidence of autoimmunity and activation of immunity will be discussed. In addition the talk will highlight the pathways and mechanisms of tissue damage that results in sight‐threatening disease.Traditionally, despite active immune regulatory mechanisms operative within the ocular environment, inflammation still occurs. Activated antigen and non‐antigen specific T cells are generated in uveitis. The interplay with innate immunity and in particular cells of myeloid lineage both systemically and within the local environment dictate the severity and extent of pathology we observe. The understanding of immune responses during the uveitis open many avenues to potential novel immunotherapies that not only suppress inflammation but attempt to redress immune balance, tolerance and local homeostasis within ocular tissues.
The role of Meis genes is lens and retina development
Authors: Kozmik Z. , Antosova B.
Published: September 2017

While significant insights into the functional role of some transcription factors for eye formation have been accomplished, much less is known about the intricate wiring of the gene regulatory network (GRN) that controls the earliest stages of eye development. Here we focus on the role of Meis homeobox genes in mammalian lens and retina development. It is well established that Pax6 represents a key node in the GRN governing lens induction. Meis1 and Meis2 were proposed to be essential upstream regulators of Pax6 during lens morphogenesis. This hypothesis is corroborated by the fact that Meis1/Meis2 double‐deficient mice phenocopy Pax6 mutant mice at the lens placode stage. We demonstrate that Meis1 and Meis2 homeoproteins regulate Pax6 gene via two redundant (shadow) enhancers, EE and SIMO. Simultaneous genetic ablation of EE and SIMO enhancers demostrates their requirement for lens induction. Using conditional gene targeting in mice at optic cup stage we provide evidence of a role of Meis genes in the process of retina development. Combined, our data indicate that Meis1 and Meis2 genes are required for normal lens and retina development.
Genes and regulation of eye development
Authors: Semina E. , Sorokina E. , Muheisen S. , Hendee K. , Weh E. , Reis L.
Published: September 2017

Developmental ocular disorders have a strong genetic component. Our laboratory has been applying whole exome sequencing (WES) to identify novel human genes associated with abnormal eye development for many years. WES provides the opportunity to simultaneously screen coding regions of all genes in order to identify causative mutations. While this comprehensive examination greatly improves the chances of mutation detection, interpretation of the large volume of data is complicated. Since many developmental processes are conserved in vertebrates, studies of zebrafish orthologs of human disease genes can provide critical information about their embryonic function and determine other genes involved in the same pathway. With this, studies of zebrafish mutants help to identify an additional pool of relevant gene candidates for corresponding human phenotypes. Moreover, zebrafish model allows for rapid in vivo testing of promising human variants by overexpression and/or rescue experiments. Our utilization of this approach in studies of zebrafish orthologs of several transcription factors, such as PITX2, PITX3 and FOXE3 associated with Axenfeld‐Rieger syndrome, anterior segment dysgenesis, cataracts and microphthalmia, will be presented.
Omega‐3 fatty acids supplementation: therapeutic potential in a mouse model of Stargardt's disease
Authors: Georgiou T. , Prokopiou E. , Kolovos P. , Kalogerou M. , Neokleous A. , Nicolaou O. , Sokratous K. , Kyriacou K.
Published: September 2017

Purpose

To evaluate the therapeutic effects of omega‐3 (ω‐3) fatty acids in the ABCA4‐/‐ model of Stargardt's disease. Monitoring the blood level of eicosapentaenoic acid (EPA) and arachidonic acid (AA), served to adjust the treatment dosage (AA/EPA = 1‐1.5).

Methods

Eight‐month‐old mice were allocated to different groups: A) wild type (129S1), B) ABCA4‐/‐ untreated, C) ABCA4‐/‐ treated with ω‐3. Treatment was daily administered by gavage for 3 months. Eye cups from each group were histologically examined using confocal and transmission electron microscopy. Proteomic analysis and N‐retinylidene‐N‐retinylethanolamine (A2E) quantification was carried our using liquid chromatography mass spectrometry (LC‐MS/MS). Blood and retinal fatty acids analysis was performed using gas chromatography (GC).

Results

A significant decrease in melanolipofuscin granules in the retinal pigment epithelium (RPE) was observed in the treatment group, in parallel with a reduction in A2E level, a major component of RPE lipofuscin. Proteomic analysis indicated decreased levels of complement 3 (C3) in the treated group. The outer nuclear layer (ONL) thickness was significantly greater in group C (75.66 ± 4.8 μm), compared to group A (61.40 ± 1.84 μm) and B (56.50 ± 3.24 μm). Increased EPA and decreased AA levels were observed in both blood and retinas in the treatment group.

Conclusions

Supplementation with ω‐3 fatty acids (when AA/EPA = 1‐1.5) suggests a protective mechanism in the ABCA4‐/‐ animal model of Stargadt's disease.
Nanostructured hydroxyapatite used as an augmenting material to expand the orbit
Authors: Popa Cherecheanu A. , Istrate S. , Iancu R. , Popescu M. , Bastian A. , Ciuluvica R.
Published: September 2017

Purpose

To test the nanostructured hydroxyapatite as an augmenting material to expand the orbit after initial placement of a porous implant.

Methods

Nanostructured hydroxyapatite powder mixed with blood was surgically introduced in the scleral sac of 6 eviscated rabbits (Oryctolagus cuniculus europaeus) after previous placing of a porous hydroxyapatite integrated implant (proved by CT scan). CT scans were done at 1, 2 and 3 months postsurgical to demonstrate the integration of the injected material in the porous implant. The scleral sac wes removed after 3 months and histopathologic examination plus CD31 +  (as a marker of neovascularization and osteosinthesis) were performed.

Results

CTscans performed at 1, 2 and 3 months postoperatively and the rapid development of CD31 +  osteoclasts and vascular tissue pleade for a good integration of the new material in the previous porous hydroxyapatite implant.

Conclusions

This method can be used for enhancing the volume of a small scleral sac with a previous hidroxyapatite implant. Further studies should be performed.
OCT‐A: Diagnosis and management of surgical macular pathologies
Authors: Pournaras C.
Published: September 2017

The aim of the coarse is to demonstrate, using OCT‐A, the most frequent clinical issues for various pathologies of the macular vitreoretinal interface et their typical evolution following an appropriate surgery.

OCT angiography (OCT‐A) enable the simultaneous monitoring of the macular anatomic changes, the macular micro‐circulation and flow, allowing the correlation of the functional to the anatomical outcomes during the progression of the anatomical damage as well as their recovery following surgery.

OCT‐A indicate and improvement or the resolution the macular contour, associated with the improvement of the perifoveal capillary arcade distortion. The preoperative intraretinal cystoid changes, associated to ischemic changes mainly of the deep capillaries plexus, regresses following surgery. In addition the regression of the associated cystoid inner retinal cystoid changes is associated with the recovery of the superficial and deep capillaries integrity.

OCT‐A evaluation following vitrectomy, demonstrated the transient anatomical and ischemic changes of the inner retina, reversible following an appropriate surgery.
Is it feasible to treat?
Authors: Levin L.
Published: September 2017

If we assume that a neuroprotective therapy for glaucoma exists and is truly effective, what are the barriers to making it available to clinicians for use in their patients? This talk will discuss four specific processes that need to take place before neuroprotection can become clinically available:

1) Presence of a neuroprotective effect in a well‐designed and powered proof‐of‐concept clinical trial.

2) Demonstration of a clinically meaningful effect in two or more Phase 3 clinical trials acceptable to regulatory agencies.

3) Pricing and clinical effectiveness that meets national/regional standards for incremental cost‐effectiveness ratios.

4) Integration of clinical effectiveness, patient acceptability, and adverse effect profiles into care guidelines that are promulgated by ophthalmology specialty societies and key opinion leaders.

The critical issues underlying each of these four steps will be analyzed.
Insult dependent oxidative‐induced cell death
Authors: Osborne N.
Published: September 2017

Mitochondrial dysfunction caused by inhibition of oxidative phosphorylation results both in reduced ATP production and oxidative stress because of a significant elevation of ROS. In the reported studies oxidative stress to cultures of cells (RGC‐5 cells) was initiated by inhibition of mitochondrial phosphorylation (blue light, sodium azide) or by use of hydrogen peroxide known to induce superoxide through the activation of cellular NADPH oxidase. While all insults resulted in cell death and attenuated by the antioxidant EGCG differences occurred in various measured biochemical parameters implicated in cell death. For example, blue light‐induced apoptosis was attenuated by the presence of the mitochondrial the uncoupler M3778 but potentiated by the presence of cobalt. In contrast, hydrogen‐peroxide‐induced apoptosis was unaffected by M3778 but attenuated by cobalt. Also, blue light‐induced cell death occurs via necroptosis, in that it was inhibited by necrostatin‐1 and was caspase‐independent. This was not the case for sodium azide, where the death process was caspase‐dependent, occurred via apoptosis and was unaffected by necrostatin‐1.
Fluorescence lifetimes of drusen in age‐related macular degeneration
Authors: Dysli C. , Fink R. , Wolf S. , Zinkernagel M.S.
Published: September 2017

Purpose

Fluorescence Lifetime Imaging Ophthalmoscopy (FLIO) allows non‐invasive in vivo measurement of fluorescence lifetimes of natural fluorophores of the retina upon excitation with a laser light. FLIO in different retinal diseases showed that the technique is useful to detect early metabolic changes and can differentiate between retinal deposits.

The aim of this study was to characterize fluorescence lifetimes in retinal drusen due to age‐related macular degeneration (AMD).

Methods

FLIO imaging was performed using a Fluorescence Lifetime Imaging Ophthalmoscope (Heidelberg Engineering, Germany). Autofluorescence was elicited with a 473 nm laser and decay times were measured in a short (498–560 nm) and in a long (560–720 nm) spectral channel. Fluorescence lifetime data was compared with autofluorescence intensity images, optical coherence tomography (OCT) and color fundus images. Soft drusen and reticular pseudodrusen were analysed and compared to an age matched control group.

Results

64 eyes from 64 patients with AMD and retinal drusen (age: mean ± SD 78 ± 8.5 years) were investigated and compared to 20 age matched healthy controls. Mean fluorescence lifetime in patients with AMD was significantly prolonged compared to the healthy control eyes (mean ± SEM; SSC 486 ± 18 ps vs 332 ± 11 ps, p > 0.0001; LSC: 493 ± 9 ps vs 382 ± 17 ps, p > 0.0001). Areas of drusen featured a broader range of fluorescence lifetime values. Long lifetimes were identified in areas of atrophy and intraretinal hyperreflective deposits. Areas of short lifetime corresponded to deposits within the photoreceptor outer segment band.

Conclusions

Mean retinal autofluorescence lifetimes in AMD were significantly prolonged compared to healthy control eyes. Intraretinal deposits caused prolonged lifetimes whereas deposits in the area of the outer photoreceptor segments lead to short fluorescence lifetimes.
Structural Bscan OCT correlation with OCT angiography biomarkers of activity in neovascular age related macular degeneration
Authors: Ambresin A. , Mantel I. , Bergin C. , Naso S.
Published: September 2017

Purpose

Fluid on structural B scan OCT is currently the most important sign among all imaging modalities of neovascular AMD. Recently, OCT angiography biomarkers of activity of nAMD have been described. The aim of the study is to compare the appearance of OCT angiography of eyes with wAMD with intraretinal or subretinal fluid against those without fluid on Bscan OCT.

Methods

Retrospective cross sectional study.Fluid on Bscan OCT was classified as intraretinal fluid such as cysts and subretinal fluid as fluid under the retina. OCT angiography biomarkers of activity of nAMD were presence of secondary small vessel branching, peripheral arcades with anastomoses and presence of a dense net (DN). Inclusion criteria were: presence of neovessels on OCT angiography and sufficent images quality on both modalities. Presence of degenerative cysts and/or PED was an exclusion criteria.

Results

59 eyes of 55 patients naive or treated for wet AMD were included (average age: 80 (SD ±7), min‐max: 63‐95). Subretinal fluid was present in 20% eyes (12/59), intraretinal fluid in 25% (15/59), a combination of both in 10% (6/59) and absence of subretinal or intraretinal fluid in 44% (26/59). DN was present in 30% (18/59), LN in 69% (41/59). Dense net was present in statistically significantly more of the uncontrolled eyes 45% (15/33) (p=0.009, fisher exact test.) Peripheral arcades with anastomoses were evident in 46% (12/26) of controlled eyes and 61% (20/33) of uncontrolled eyes. Small vessel branching was evident in 0% (0/26) of the controlled eyes and in 36% (12/33) uncontrolled eyes.

Conclusions

Dense net are significantly more prevalent in eyes with uncontrolled AMD; small vessel branching and peripheral loops are also more prevalent in these eyes. This suggest that OCT‐A provides imaging of the morphology of choroidal CNV in AMD patients associated with disease activity.
Evidence for successful treatment in glaucoma: who to treat and when?
Authors: Gandolfi S.
Published: September 2017

Treating glaucoma successfully means avoiding the occurrence of visual disability in the individual patient. Since the rate of vision loss can be extremely variable among patients, an accurate phenotyping is essential in planning the proper therapy for each patient. Besides, the recent inclusion of patients’ reported outcomes (PROs), in the glaucoma therapeutic algorithm, is increasing the complexity of the scenario.

This presentation will focus, then, on (a) the available tools to phenotype the glaucoma patient(s), (b) the criteria for assessing the intensity and the timing of different treatment modalities and (c) the state‐of‐the‐art in the evaluation of PROs in glaucoma.
Over‐interpreting white dots
Authors: Srivastava S.
Published: September 2017

The white dot syndromes are a group of inflammatory chorioretinopathies of unknown etiology which have in common a unique and characteristic appearance of multiple yellow‐white lesions affecting multiple layers of the retina, retinal pigment epithelium (RPE), choriocapillaris, and the choroid.

They also have overlapping clinical features. For example, multifocal choroiditis and punctate inner choroidopathy target the same essential structures in the same phenotypic manner and, when active, are treated the same way, and thus there seems to be limited clinical utility in trying to differentiate them.

Transitions between the individual diseases have also been described, and when a prolonged progressive clinical course and widespread distribution of lesions is present, the term “relentless” is now applied.

Neither the trigger mechanism nor the pathogenetic development is known with certainty for any of these diseases. Immunological reactions to previous viral infections coupled with a genetic predisposition seem to be a common denominator. Some of these conditions share an association with systemic infectious diseases.
Choosing the wrong imaging modalities
Authors: Francesco P.
Published: September 2017

The diagnosis of uveitis often requires careful clinical examination of the peripheral retina. Ultra‐wide‐field angiography could prove useful in the clinical evaluation and treatment such patients. However, its use is often restricted to intermediate and posterior uveitis. There are instances in which ultra‐wide‐field FA can be paramount in evaluating the peripheral angiographic expression of the retina in anterior uveitis.

Fundus autofluorescence (FAF) is a novel noninvasive imaging technique that assess the health and viability of the RPE/photoreceptor complex. Characteristic fundus autofluorescence patterns have been described in eyes affected by inflammation of the posterior segment, and these patterns have provided insights into the pathogenesis of posterior uveitis entities. In addition, preliminary data indicate that fundus autofluorescence characteristics may serve as markers of disease activity, allow prediction of visual prognosis, and may help determine the adequacy of therapy.

Indocyanine green angiography (ICGA) provides the clinician with a powerful adjunctive tool in choroidal inflammatory disorders. It still needs to be determined if ICGA can prove to be a follow up parameter to evaluate disease progression.
Efficacy of a RARγ selective agonist eye drop formulation on improvement of tear production and corneal fluorescein staining in the BTX‐B mouse model of dry eye disease
Authors: Lemire I. , Harvey M. , Grogan D. , Desjardins C.
Published: September 2017

Purpose

Multiple studies have demonstrated the potential beneficial effects of retinoids on dry eye disease. Retinoic acid receptors (RAR) α, β and γ are widely distributed in ocular tissues. In vitro data suggest that the beneficial effects of retinoids on ocular health are mediated via RARγ (Kimura 2015). The purpose of this study was to evaluate the in vivo effects of palovarotene (PVO), a RARγ selective agonist, in a dry eye animal model.

Methods

The efficacy of a PVO ocular formulation was tested in the botulinum toxin type B (BTX‐B) mouse model which involves injection of BTX‐B into the lacrimal gland of the right eye with contralateral control. Eye drop formulations at 3 doses of PVO (low, mid, high) with vehicle, were administered by daily topical instillation for 28 consecutive days and compared to current standard of care, Restasis®. Endpoints included ophthalmic examinations, tear production measurement (TPM) by phenol red‐impregnated cotton threads, corneal fluorescein staining (CFS) using a scoring system of 0 to 4, and histopathology.

Results

At Day 28 post BTX‐B injection, the mean (SD) TPM and CFS score in the untreated group were 1.5 (0.3) mm and 2.0 (0.7), respectively. In comparison, PVO eye drop treatment significantly improved dry eye signs (p<.0001) at all doses tested – TPM was increased by 132, 166 and 188% at low, mid and high dose, respectively, while CFS score was decreased by 70, 85 and 85%, respectively. PVO treatment effects were greater than Restasis® (113% increase in TPM, 50% decrease in CFS). All PVO doses were tolerated for 28 days. Histopathology evaluation revealed no effects on the Meibomian glands.

Conclusions

These results indicate a beneficial therapeutic effect of PVO in the BTX‐B animal model and support further development of a PVO ocular formulation for the treatment of human dry eye disease.
Graft blues: case report
Authors: Thuret G. , Marcon A. , Perillat N. , Jullienne R. , Garcin T. , He Z. , Peoc'H M. , Gain P.
Published: September 2017

Purpose

To describe unusual inadvertent persistent staining of stromal structures by trypan blue (TB) after penetrating keratoplasty (PK) and Descemet membrane endothelial keratoplasty (DMEK) performed in patients suffering from lattice corneal dystrophy (LCD)

Methods

Case report of patients suffering from LCD and having received intracamerular TB. Series 1: retrospective study on 85 consecutive triple procedures (PK+cataract+IOL) performed by a single surgeon (PG) in a tertiary care university hospital during 7 years. 0.4% TB was systematically used. Series 2: case report of a DMEK (stained with TB) performed after a late endothelial failure of a previous PK realized 5 years before for advanced LCD. Six weeks after DMEK, the patient was hospitalized for acute rejection that appeared unusual

Results

Series 1: Only patients with LCD (n = 18 eyes in 17 patients) presented an isolated intense blue staining of the graft host interface (blue ring). It persisted during 18‐24 months and had no consequence. Series 2: during the acute rejection, intense blue staining was observed in spindle shaped small structures located in the first third of the whole stroma. The DMEK was partially detached. Stromal edema almost completely resolved after intense steroid therapy but the stromal blue staining persisted (9 month follow‐up) with impaired vision.

Conclusions

Abnormal amyloid protein characterizing LCD can be stained by TB in vivo. In PK it stains a ring in the recipient stroma and has no consequence. However, during DMEK performed after a PK, TB may also stain the protein accumulated in the graft stroma during slow recurrence of the disease. TB‐stained DMEK should therefore be avoided in late endothelial failure of a PK initially performed for LCD.
Modulation of Muller cell membrane organization by 24S‐hydroxycholesterol
Authors: Gabrielle P.H. , Gambert S. , Masson E. , Leger‐Charnay E. , Ferrerro A. , Vannier A. , Gendrault C. , Lachot M. , Creuzot‐Garcher C. , Bon A. , Gregoire S. , Leclere L. , Martine L. , Lucchi G. , Truntzer C. , Bretillon L.
Published: September 2017

Purpose

Muller glial cells (MGC) are involved in the retinal homeostasis and they are sustaining cholesterol homeostasis. A lack or an excess of cholesterol in neurons can cause neuro‐degeneration. Cholesterol is eliminated from the retina mainly as 24S‐hydroxycholesterol (24S‐OHC). 24S‐OHC is produced by 24S‐hydroxylase (CYP46A1) in retinal ganglion cells (RGC) and is overexpressed during glaucoma. the objective of this study was to determine whether 24S‐OHC triggers MGC membrane dynamics involving lipid rafts and contribute to gliosis at early and late time points.

Methods

MGCs were grown in vitro from retinas of young rats. They were treated with 24S‐OHC (10 μM) for 2 min or 6h. Lipid rafts of MGC membranes were obtained after 1% lubrol lysis and 20h‐ultracentrifugation at 180,000 g in a sucrose gradient. The expression of caveolin‐1, Flotillin‐1, GFAP, Vimentin, Connexin‐30, Connexin‐43, phosphorylated and non phosphorylated p38 and p42‐44 MAPK was analyzed by Western‐blotting. High‐performance liquid chromatography coupled with mass spectrometry (LC‐MS) was used to estimate the levels of ganglioside GM3 (monosialodihexosylganglioside), and protein pathways were analyzed by nanoLC‐MS/MS in raft and non raft fractions from MGCs after treatment with 24S‐HOC.

Results

Cholesterol, sphingomyelin and saturated fatty acids C15:0, C16:0, C17:0 and C18:0 were enriched in the rafts fractions in MGCs. Structural proteins, Caveolin‐1 and flotillin‐1, and functional proteins, Connexin‐30 and ‐43, were localized in the MGCs rafts. Ganglioside GM3 showed a characteristic enrichment in the raft fractions. Protein implicated in adhesion or oxidative stress pathways in the raft and non‐raft fractions were up and down‐regulated by treatment with 24S‐OHC.

Conclusions

Our data showed that 24S‐OHC induced early changes in protein distribution in raft microdomains.
Critical points in scientific retinal imaging
Authors: Felszeghy S. , Viiri J. , Kettunen M. , Kositinen A. , Kai K.
Published: September 2017

Images obviously are one of the most important category among the available data in everyday clinical ophthalmology and the visualization of different layers of retina. In experimental eye research it also play pivotal role in understanding of different biological processes of retina. This talk will focus on image capturing and analysis of mouse retina with different genetic background. In our project, we have focused on retinal pigmented epithelium (RPE) obtained from different degenerative mouse models mimicking cellular changes observed in Age‐related Macular Degeneration (AMD). The 2D and 3D of RPE images were acquired through different types of microscopy. We deliver results on modern methods of absolute nucleic acid or protein quantification (q‐PCR, WB, ELISA), but highlight the importance of imaging of the bio‐molecular processes by a range of powerful imaging weapons (stereo/light/confocal/EM/microMRI). We have carried out do novo, in and ex vivo analyzis at micro and ultrastructural level to describe the molecular fingerprints of mouse RPE features in AMD.
Different light spectra to better visualize retinal detail
Authors: Uusitalo H. , Lensu L. , Hauta‐Kasari M.
Published: September 2017

Retinal imaging provides a unique opportunity to directly visualize central nervous system and its blood vessels and analyze their structural and functional changes with modern technologies. Retinal imaging has been used for the diagnosis and follow‐up of various eye diseases like glaucoma and age‐related macular degeneration. Also in many systemic diseases like diabetes retinal imaging has been developed as the state of art diagnostic tool. It is one of the most sensitive tools to detect diabetic complications and screening programs to detect retinal complications has been set‐up worldwide. To provide a proper care for increasing numbers of patients is an enormous burden to the health care and will need a new thinking and technologies. Personalized health care, individualized follow‐up and automated self‐testing technologies will neccesary tools to beat these challenges. Re‐engineering retinal imaging with photonics and computational science is a multidisciplinary approach based on spectral imaging and analysis of human retina. It has realized a spectral eye fundus imaging device, spectrally tunable light source device and spectral image database of eye fundus images with clinical expert markings.
Anterior to posterior “tour” of ocular disease processes
Authors: Van Ginderdeuren R.
Published: September 2017

In this part of the course the most important pathology of the internal eye structures will be explained from front to back side of the eye. The malignant tumours of adults (melanoma, metastasis, lymphoma) and children (retinoblastoma) will be covered, but also the differential diagnosis and important consequences of non‐malignant diseases (sympathetic ophthalmia). Newer technologies can be incorporated in the pathologic examination which can have far reaching consequences for the patients. Enucleation and the difference with evisceration specimens will be discussed. Newer methods for fine needle aspiration biopsy (in solid tumours) and vitreo‐retinal biopsies in malignant and inflammatory processes will be explained.
Pre‐receptoral filters in the eye and their effect on vision
Authors: Barbur J.
Published: September 2017

Advanced Vision and Optometric Tests (www.city.ac.uk/avot) were employed to study changes in macular pigment optical density and visual performance following three months of daily supplementation with carotenoids. The subject investigated was earlier diagnosed with type 2 diabetes, but recovered through diet and exercise, to the point of being free of diabetes.

Before supplementation, the tests revealed borderline performance for photopic and mesopic visual acuity (VA), functional contrast sensitivity (FCS) and red / green (RG) colour vision. The subject's thresholds for yellow‐blue (YB) colour vision and rod sensitivity were, however, just above normal, age‐matched limits.

Post supplementation, the results revealed increased macular pigment levels in the central retina. In addition, the results also revealed significant improvements in several, other aspects of vision, particularly at lower light levels. VA and FCS improved, but this outcome depended on whether the stimuli were of positive or negative luminance contrast. Rod and cone mediated vision showed significant changes with large improvements in rod sensitivity. Thresholds for detection of RG and particularly YB colours were also significantly lower post‐supplementation.
Circadian regulation of outer segments phagocytosis by RPE cells: more complexity than meets the eye
Authors: Nandrot E.
Published: September 2017

Daily clearance of aged photoreceptor outer segments (POS) extremities is one of the main functions of retinal pigment epithelial (RPE) cells. Absence or deregulation of this process leads to early‐onset or age‐related retinal pathologies. Despite the permanent tight contact between POS and RPE microvilli, phagocytosis occurs only once a day. Over the years, the main molecules implicated in POS tethering and subsequent internalization have been identified. POS tips expose phosphatidylserines on the outer membrane portion to be cleared. Timely recognition of POS tips and phagocytosis synchronization are mediated by the MFG‐E8–alphavbeta5 integrin ligand–receptor couple. This triggers intracellular signaling pathways leading to the activation of the MerTK internalization receptor. While this draws a sequential scheme of events, things are far from being that simple. Recently new roles for MerTK and implication of new receptors have been suggested. In contrast to macrophages that share a similar phagocytic machinery, the continuous contact between both cell types requires the strict regulation of phagocytosis launch and stop to ensure lifelong retinal function. Thus, at this point the puzzle is about to get much more complicated.
Leber's hereditary optic neuropathy: the neurologist point of view
Authors: Carelli V. , La Morgia C.
Published: September 2017

Leber's hereditary optic neuropathy (LHON) is a neurodegenerative disorder that selectively affects the retinal ganglion cells (RGCs), consequently leading to optic atrophy and loss of central vision. The primary cause is a metabolic dysfunction of mitochondria due to mitochondrial DNA mutations affecting the respiratory complex I. Despite the metabolic defect is systemic and measurable in post‐mitotic tissues, the disease is usually limited to the RGCs and leads to profoundly impaired vision. However, adjunctive neurological features may be present in a subset of LHON patients defining the so‐called “plus” variant phenotype. The most frequent neurological features may be a multiple‐sclerosis‐like disease, basal ganglia involvement with dystonia or spastic dystonia and bilateral striatal necrosis, myoclonus, parkinsonism, peripheral neuropathy, migraine and sensorineural hypoacusia. Neurological exam, brain MRI, MR‐spectroscopy and other ancillary exams may be needed to well characterize the “plus” phenotype of LHON patients. The genetic bases of these LHON “plus” cases are currently unclear, but private mtDNA mutations and nuclear genetic variants may be relevant.
Uveal melanoma biopsy or not (against)
Authors: Caujolle J.P.
Published: September 2017

Biopsy for diagnostic or prognostic purposes are still controversial issues. The arguments depend on the biopsy technic used and the reason why? Concerning the biopsy, different complications could occur such as intravitreal or tumoral hemorrhages, retinal detachment, cataract, endophthalmitis and the last one tumoral cells spreading. All these complications must be taken into account concerning the possible benefits of a biopsy. In ocular oncology units, the diagnosis accuracy reported with noninvasive technics is over 98% with big tumors requiring enucleation. This rate could probably be similar with medium tumors. However in routine, difficult diagnosis concern small uveal tumors less than 3 mm in thickness. In a few countries physicians perform biopsy for such tumors but not in France. Our management is periodical follow up until a proof of growth. That is why, we do not really need a biopsy to diagnose an uveal melanoma!!! On one hand, we are not able to make a prognosis in 12 to 24% of cases of cytogenetic analysis. On the other hand, nowadays, we do not have a successful proven adjuvant therapy to propose to our high risk patients. How many patients depending on their origins would like to know whether they will die or not?
Endoresectiopn of uveal melanoma without radiotherapy
Authors: Damato B.
Published: September 2017

Endoresection of choroidal melanoma without neoadjuvant radiotherapy is controversial because of concerns about iatrogenic local tumor seeding and systemic metastases. However, several authors have reported that local tumor recurrence and seeding are rare. Further, there is no evidence that metastatic disease is increased following endoresection. Recent insights into uveal melanomas indicates that metastasis depends on the genetic tumor profile and not on surgical interventions. It would seem, therefore, that the large majority of patients experiencing ocular morbidity from neoadjuvant radiotherapy are doing so unnecessarily. This problem persists because those favoring neoadjuvant radiotherapy are skeptical of the results published by authors who hold a different view from their own. There is scope for randomized clinical trials of endoresection with and without neoadjuvant radiotherapy. If the required equipoise for such trials is lacking, an ethical way should be found to allow independent inspectors to review original patient case notes and verify all the scientific claims that neoadjuvant radiotherapy is unnecessary.
Comparison of four technics of surface roughness assessment of corneal lamellar cuts
Authors: Gain P. , Jumelle C. , Hamri A. , Egaud G. , Mauclair C. , Reynaud S. , Dumas S. , Pereira S. , Thuret G.
Published: September 2017

Purpose

Surface roughness of corneal lamellar cuts is a crucial quality criterion, because smother is the cutting plan, better will be the visual recovery. Nevertheless, despite several studies published in the last years, it remains difficult to reliably quantify the differences between a microkeratome (MKT) and a femtosecond laser (FSL). A possible reason could be that the different studies employed different technics to assess surface roughness. Aim

To directly compare 4 microscopy technics of surface roughness assessment of corneal lamellar cuts.

Methods

Stromal lamellae were cut in organ cultured human corneas with a Moria MKT or a FSL. Environmental scanning electron microscopy (eSEM), standard SEM and chromatic confocal microscopy (CCM) were first used to verify that dehydration and metallization of samples did not artefactually alter the surface state. Three 3 quantitative methods (Atomic Force Microscopy (AFM), CCM and Focus‐Variation Microscopy (FVM)) were then compared using the same stromal lamellae. Lamellae with low or high surface roughness were selected with sSEM. The Bowman membrane was used as a smooth control. Roughness quantification was done with Mountains, a validated software.

Results

Compared to eSEM, sSEM provided better image contrast allowing better visualization of the surface roughness. Dehydration and metallization did not change the surface appearance with eSEM and sSEM, and did not modify the roughnesses measured by CCM. Only CCM and FVM were able to reveal significant differences between lamellae of different roughness. CCM allowed acquisition of larger areas, allowing a better characterization of heterogeneous surfaces.

Conclusions

CCM seems to be particularly suitable for quantifying the roughness of cutting plans obtained in the cornea.