Inhibition of cholesterol recycling impairs cellular PrPSc propagation

The infectious agent in prion diseases consists of an aberrantly folded isoform of the cellular prion protein (PrPc), termed PrPSc, which accumulates in brains of affected individuals. Studies on prion-infected cultured cells indicate that cellular cholesterol homeostasis influences PrPSc propagation. Here, we demonstrate that the cellular PrPSc content decreases upon accumulation of cholesterol in late endosomes, as induced by NPC-1 knock-down or treatment with U18666A. PrPc trafficking, lipid raft association, and membrane turnover are not significantly altered by such treatments. Cellular PrPSc formation is not impaired, suggesting that PrPSc degradation is increased by intracellular cholesterol accumulation. Interestingly, PrPSc propagation in U18666A-treated cells was partially restored by overexpression of rab 9, which causes redistribution of cholesterol and possibly of PrPSc to the trans-Golgi network. Surprisingly, rab 9 overexpression itself reduced cellular PrPSc content, indicating that PrPSc production is highly sensitive to alterations in dynamics of vesicle trafficking

Identification of an Intracellular Site of Prion Conversion

Prion diseases are fatal, neurodegenerative disorders in humans and animals and are characterized by the accumulation of an abnormally folded isoform of the cellular prion protein (PrPC), denoted PrPSc, which represents the major component of infectious scrapie prions. Characterization of the mechanism of conversion of PrPC into PrPSc and identification of the intracellular site where it occurs are among the most important questions in prion biology. Despite numerous efforts, both of these questions remain unsolved. We have quantitatively analyzed the distribution of PrPC and PrPSc and measured PrPSc levels in different infected neuronal cell lines in which protein trafficking has been selectively impaired. Our data exclude roles for both early and late endosomes and identify the endosomal recycling compartment as the likely site of prion conversion. These findings represent a fundamental step towards understanding the cellular mechanism of prion conversion and will allow the development of new therapeutic approaches for prion diseases

Anterior-segment morphology and corneal biomechanical characteristics in pigmentary glaucoma

Annemarie Klingenstein, Marcus Kernt, Florian Seidensticker, Anselm Kampik, Christoph HirneissDepartment of Ophthalmology, University of Munich Hospital, Ludwig-Maximilians University, Munich, GermanyPurpose: The aim of the study reported here was to evaluate characteristics of the anterior-segment via anterior-segment optical coherence tomography (AS-OCT) and corneal biomechanical properties using an ocular response analyzer and their changes by peripheral laser iridotomy (PI) in patients with pigmentary glaucoma (PG).Materials and methods: Seventeen eyes with PG were included consecutively. AS-OCT and ocular response analyzer measurements were taken before and 3 months after PI. Baseline morphology and change in morphology were analyzed by correlation and multiple linear regression analysis. The main parameters assessed were anterior-chamber (AC) angles and volume as well as corneal hysteresis (CH) and corneal resistance factor.Results: AC angles were found to have decreased significantly in each quadrant after PI (P<0.001), with the highest effect seen in the temporal quadrant, which decreased from 57.0°±9.6° to 44.1°±5.2° (± standard deviation). Mean AC volume decreased significantly from 213.1±36.4 to 187.0±23.4 mm3 (P<0.001). CH and corneal resistance factor did not change after PI. CH was found to correlate with the preoperative superior and inferior angle width (Spearman's rho 0.553 and 0.615, respectively, P<0.05). Biomechanical parameters showed no predictive value on the change of AC angles or volume.Conclusion: PI in eyes with PG results in a highly significant reduction in the AC angles and volume as visualized by AS-OCT, with the largest effect seen in the temporal quadrant. CH is strongly positively correlated with the superior and inferior preoperative AC angles, emphasizing the importance of the biomechanical properties of the cornea for glaucoma pathogenesis in PG, but corneal biomechanical properties cannot predict PI-related AC changes.Keywords: anterior-segment optical coherence tomography, ocular response analyzer, corneal hysteresi

Anti-VEGF treatment and peripheral retinal nonperfusion in patients with central retinal vein occlusion

Kaveh Abri Aghdam,1,* Lukas Reznicek,2,* Mostafa Soltan Sanjari,1 Annemarie Klingenstein,2 Marcus Kernt,2 Florian Seidensticker2 1Department of Ophthalmology, Eye Research Center, Rassoul Akram Hospital, Iran University of Medical Sciences, Tehran, Iran; 2Department of Ophthalmology, Ludwig-Maximilians University, Munich, Germany *These authors contributed equally to this work Purpose: To evaluate the association between the size of peripheral retinal nonperfusion and the number of intravitreal ranibizumab injections in patients with treatment-naïve central retinal vein occlusion (CRVO).Methods: Fifty-four patients with treatment-naïve CRVO and macular edema were included. Each patient underwent a full ophthalmologic examination including optical coherence tomography imaging and ultrawide-field fluorescein angiography. Monthly intravitreal ranibizumab injections were applied according to the recommendations of the German Ophthalmologic Society. Two ophthalmologists quantified the areas of peripheral retinal nonperfusion (group 1= less than five disc areas, group 2= more than five disc areas). Correlation analyses between the size of nonperfusion with best-corrected visual acuity, central subfield thickness, and the number of intravitreal injections were performed.Results: Best-corrected visual acuity improved significantly after intravitreal injections (P<0.001, both groups). Final central subfield thickness after treatment did not significantly differ between both groups (P=0.92, P=0.96, respectively). Mean number of injections in group 1 and group 2 was 4.12±2.73 and 9.32±3.84, respectively (P<0.001). There was a significant positive correlation between areas of nonperfusion and the number of injections in each group. (R=0.97, P<0.001; R=0.94, P<0.001, respectively).Conclusion: Peripheral retinal nonperfusion in patients with CRVO correlates significantly with the number of needed intravitreal ranibizumab injections. Ultrawide-field fluorescein angiography is a useful tool for detection of peripheral retinal ischemia, which may have direct implications in the diagnosis, follow-up, and treatment of these patients. Keywords: angiography, central retinal vein occlusion, non-perfusion, retina, wide-fiel