Three-Dimensional Topographic Angiography in Chorioretinal Vascular Disease

PURPOSE. To evaluate a new angiographic technique that offers three-dimensional imaging of chorioretinal vascular diseases. METHODS. Fluorescein (FA) and indocyanine green angiography (ICGA) were performed using a confocal scanning laser ophthalmoscope. Tomographic series with 32 images per set were taken over a depth of 4 mm at an image frequency of 20 Hz. An axial analysis was performed for each x/y position to determine the fluorescence distribution along the z-axis. The location of the onset of fluorescence at a defined threshold intensity was identified and a depth profile was generated. The overall results of fluorescence topography were displayed in a gray scale-coded image and three-dimensional relief. RESULTS. Topographic angiography delineated the choriocapillary surface covering the posterior pole with exposed larger retinal vessels. Superficial masking of fluorescence by hemorrhage or absorbing fluid did not preclude detection of underlying diseases. Choroidal neovascularization (CNV) appeared as a vascular formation with distinct configuration and prominence. Chorioretinal infiltrates exhibited perfusion defects with dye pooling. Retinal pigment epithelium detachments (PEDs) demonstrated dynamic filling mechanisms. Intraretinal extravasation in retinal vascular disease was detected within a well-demarcated area with prominent retinal thickening. CONCLUSIONS. Confocal topographic angiography allows highresolution three-dimensional imaging of chorioretinal vascular and exudative diseases. Structural vascular changes (e.g., proliferation) are detected in respect to location and size. Dynamic processes (e.g., perfusion defects, extravasation, and barrier dysfunction) are clearly identified and may be quantified. Topographic angiography is a promising technique in the diagnosis, therapeutic evaluation, and pathophysiological evaluation of macular disease. (Invest Ophthalmol Vis Sci. 2001;42: 2386 -2394 C horioretinal vascular disease of the macular area (e.g., diabetic maculopathy [DMP] and age-related macular degeneration [ARMD]) are the main reasons for progressive and severe visual loss by occlusive, proliferative, and/or exudative mechanisms. 1,2 Fluorescein angiography (FA) is the classic diagnostic tool but is often compromised by masking phenomena as a consequence of the short wavelength used. Diffuse leakage of the small fluorescein molecule causes further difficulties in identifying the origin and quantifying the dynamics of leakage. Despite stereoscopic viewing systems, many lesions remain occult, and prominence and extent of exudation are evaluated only subjectively. 2,3 Indocyanine green angiography (ICGA) is effective in the near-infrared spectrum which allows improved transmission, and, mostly bound to albumin, it is thought to extravasate minimally. 5-7 Scanning laser ophthalmoscopy (SLO), with point-source illumination and optimized excitation, has further enhanced diagnostic efficacy. 9,10 The option to scan through different retinal layers is nevertheless limited to a depth resolution of approximately 300 m. It may be used, however, to obtain topographic profiles of strongly reflecting intraocular structures, such as the optic disc and the macular region. 11 Morphometric imaging of vascular structures of retina and choroid would significantly improve the diagnosis of macular disease. A novel angiographic technology, confocal topographic angiography, has been developed that allows threedimensional (3-D) documentation of vascular structures and characterization of dynamic phenomena such as perfusion and leakage. The technique of topographic image processing was applied in the FA and ICGA analyses of representative types of chorioretinal vascular disease, to document structural and dynamic changes and to evaluate the diagnostic potential of the new method. MATERIALS AND METHODS The basic topographic principle is to use a series of lateral confocal optical sections of the chorioretinal fluorescence distribution and, by introducing a smart algorithm, to extract the 3-D profile of the surface of vascular structures and related leakage. Data acquisition was achieved with a conventional confocal scanning laser angiograph. Data processing and topographic analysis were performed on a standard desktop computer, using newly developed software. The method of confocal laser scanning topography based on ICGA has been published. 12,13 Data Acquisition FA and ICGA were performed using a confocal SLO (Heidelberg Retina Angiograph; Heidelberg Engineering, Dossenheim, Germany). Infrared images were taken for optical alignment with the fovea in the center of a 30°field corresponding to a retinal area of 9 ϫ 9 mm. For FA, 5 ml of 10% fluorescein solution (Alcon Pharma GmbH, Freiburg, Germany), an argon laser emitting at 488 nm for excitation, and filters blocking transmission of wavelengths below 510 nm were used for detection. For ICGA a 50-mg solution of ICG (ICG Pulsion, München, Germany) was administered intravenously, and excitation and detection were performed, using a diode laser emitting at 795 nm and blocking filters for wavelengths below 835 nm. The diameter of the excitation beam was 10 m at the retina. The Rayleigh range of the focal beam's waist determining depth resolution was 300 m. During the early transit phase, the scanning laser was focused onto the retinal vessels and the excitation intensity was adjusted to obtain adequate illumination. An additive ϩ3-diopter (D) refractive correction was added by using the internal focus adjustment to create a preretinal initial focus for complete sectioning of elevated lesions. An early FA/ICGA series of 32 tomographic sections was taken over a depth of 4 mm, each separated From th

rAAV Engineering for Capsid-Protein Enzyme Insertions and Mosaicism Reveals Resilience to Mutational, Structural and Thermal Perturbations

Feiner R, Teschner J, Teschner K, et al. rAAV Engineering for Capsid-Protein Enzyme Insertions and Mosaicism Reveals Resilience to Mutational, Structural and Thermal Perturbations. International Journal of Molecular Sciences. 2019;20(22): 5702.Recombinant adeno-associated viruses (rAAV) provide outstanding options for customization and superior capabilities for gene therapy. To access their full potential, facile genetic manipulation is pivotal, including capsid loop modifications. Therefore, we assessed capsid tolerance to modifications of the structural VP proteins in terms of stability and plasticity. Flexible glycine-serine linkers of increasing sizes were, at the genetic level, introduced into the 587 loop region of the VP proteins of serotype 2, the best studied AAV representative. Analyses of biological function and thermal stability with respect to genome release of viral particles revealed structural plasticity. In addition, insertion of the 29 kDa enzyme β-lactamase into the loop region was tested with a complete or a mosaic modification setting. For the mosaic approach, investigation of VP2 trans expression revealed that a Kozak sequence was required to prevent leaky scanning. Surprisingly, even the full capsid modification with β-lactamase allowed for the assembly of capsids with a concomitant increase in size. Enzyme activity assays revealed lactamase functionality for both rAAV variants, which demonstrates the structural robustness of this platform technology.</jats:p

Towards Experimental Handbooks in Catalysis

The “Seven Pillars” of oxidation catalysis proposed by Robert K. Grasselli represent an early example of phenomenological descriptors in the field of heterogeneous catalysis. Major advances in the theoretical description of catalytic reactions have been achieved in recent years and new catalysts are predicted today by using computational methods. To tackle the immense complexity of high-performance systems in reactions where selectivity is a major issue, analysis of scientific data by artificial intelligence and data science provides new opportunities for achieving improved understanding. Modern data analytics require data of highest quality and sufficient diversity. Existing data, however, frequently do not comply with these constraints. Therefore, new concepts of data generation and management are needed. Herein we present a basic approach in defining best practice procedures of measuring consistent data sets in heterogeneous catalysis using “handbooks”. Selective oxidation of short-chain alkanes over mixed metal oxide catalysts was selected as an example.DFG, 390540038, EXC 2008: Unifying Systems in Catalysis "UniSysCat

Dealing with prognostic signature instability : a strategy illustrated for cardiovascular events in patients with end-stage renal disease

Background Identification of prognostic gene expression markers from clinical cohorts might help to better understand disease etiology. A set of potentially important markers can be automatically selected when linking gene expression covariates to a clinical endpoint by multivariable regression models and regularized parameter estimation. However, this is hampered by instability due to selection from many measurements. Stability can be assessed by resampling techniques, which might guide modeling decisions, such as choice of the model class or the specific endpoint definition. Methods We specifically propose a strategy for judging the impact of different endpoint definitions, endpoint updates, different approaches for marker selection, and exclusion of outliers. This strategy is illustrated for a study with end-stage renal disease patients, who experience a yearly mortality of more than 20 %, with almost 50 % sudden cardiac death or myocardial infarction. The underlying etiology is poorly understood, and we specifically point out how our strategy can help to identify novel prognostic markers and targets for therapeutic interventions. Results For markers such as the potentially prognostic platelet glycoprotein IIb, the endpoint definition, in combination with the signature building approach is seen to have the largest impact. Removal of outliers, as identified by the proposed strategy, is also seen to considerably improve stability. Conclusions As the proposed strategy allowed us to precisely quantify the impact of modeling choices on the stability of marker identification, we suggest routine use also in other applications to prevent analysis-specific results, which are unstable, i.e. not reproducible

Indicators of acute and persistent renal damage in adult thrombotic microangiopathy.

BACKGROUND: Thrombotic microangiopathies (TMA) in adults such as thrombotic thrombocytopenic purpura (TTP) and hemolytic uremic syndrome (HUS) are life-threatening disorders if untreated. Clinical presentation is highly variable and prognostic factors for clinical course and outcome are not well established. METHODS: We performed a retrospective observational study of 62 patients with TMA, 22 males and 40 females aged 16 to 76 years, treated with plasma exchange at one center to identify clinical risk factors for the development of renal insufficiency. RESULTS: On admission, 39 of 62 patients (63%) had acute renal failure (ARF) with 32 patients (52%) requiring dialysis treatment. High systolic arterial pressure (SAP, p = 0.009) or mean arterial pressure (MAP, p = 0.027) on admission was associated with acute renal failure. Patients with SAP>140 mmHg on admission had a sevenfold increased risk of severe kidney disease (OR 7.464, CI 2.097-26.565). MAP>100 mmHg indicated a fourfold increased risk for acute renal failure (OR 4.261, CI 1.400-12.972). High SAP, diastolic arterial pressure (DAP), and MAP on admission were also independent risk factors for persistent renal insufficiency with the strongest correlation for high MAP. Moreover, a high C-reactive protein (CRP) level on admission correlated with renal failure in the course of the disease (p = 0.003). At discharge, renal function in 11 of 39 patients (28%) had fully recovered, 14 patients (23%) remained on dialysis, and 14 patients (23%) had non-dialysis-dependent chronic kidney disease. Seven patients (11%) died. We identified an older age as risk factor for death. CONCLUSIONS: High blood pressure as well as high CRP serum levels on admission are associated with renal insufficiency in TMA. High blood pressure on admission is also a strong predictor of sustained renal insufficiency. Thus, adult TMA patients with high blood pressure may require special attention to prevent persistent renal failure