Assessment of choroidal thickness and volume during the water drinking test by swept-source optical coherence tomography.

OBJECTIVE: To evaluate changes in peripapillary and macular choroidal thickness and volume after the water-drinking test (WDT) using swept-source optical coherence tomography (SS OCT). DESIGN: Prospective, cross-sectional, observational study. PARTICIPANTS: Fifty-six eyes of 28 healthy volunteers. METHODS: Participants underwent a 3-dimensional optic disc and macula scanning protocol with a prototype SS OCT (Topcon, Inc., Tokyo, Japan) at baseline and 15, 30, 45, and 120 minutes after the start of the WDT. The WDT consisted of drinking 1000 ml of water within 5 minutes. Objective measurements of the choroid were obtained with automated segmentation of the choroidal boundaries. MAIN OUTCOME MEASURES: Choroidal thickness and volume. RESULTS: Mean age ± standard deviation of participants was 35.6 ± 9.1 years. Intraocular pressure (IOP) increased from 14.9 ± 2.7 mmHg at baseline to a peak of 16.8 ± 3.0 mmHg 15 minutes after the WDT (P < 0.001). Mean baseline choroidal thickness and volume were 181.3 ± 50.8 µm and 6.19 ± 1.80 mm(3), respectively, at the optic disc and 217.4 ± 43.6 µm and 7.83 ± 1.55 mm(3), respectively, at the macula. After the WDT, peripapillary and macular choroidal thickness increased by a maximum of 5.7% (P<0.001) and 4.3% (P<0.001), respectively. Choroidal volumes increased by 6.4% (P<0.001) and 3.9% (P<0.001), respectively. There was no association between change in IOP and peripapillary (P = 0.27) or macular (P = 0.09) choroidal thickness. CONCLUSIONS: Using automated segmentation of SS OCT measurements, significant increases in choroidal thickness and volume are observed after the WDT in healthy subjects.

Yeast two-hybrid, a powerful tool for systems biology.

A key property of complex biological systems is the presence of interaction networks formed by its different components, primarily proteins. These are crucial for all levels of cellular function, including architecture, metabolism and signalling, as well as the availability of cellular energy. Very stable, but also rather transient and dynamic protein-protein interactions generate new system properties at the level of multiprotein complexes, cellular compartments or the entire cell. Thus, interactomics is expected to largely contribute to emerging fields like systems biology or systems bioenergetics. The more recent technological development of high-throughput methods for interactomics research will dramatically increase our knowledge of protein interaction networks. The two most frequently used methods are yeast two-hybrid (Y2H) screening, a well established genetic in vivo approach, and affinity purification of complexes followed by mass spectrometry analysis, an emerging biochemical in vitro technique. So far, a majority of published interactions have been detected using an Y2H screen. However, with the massive application of this method, also some limitations have become apparent. This review provides an overview on available yeast two-hybrid methods, in particular focusing on more recent approaches. These allow detection of protein interactions in their native environment, as e.g. in the cytosol or bound to a membrane, by using cytosolic signalling cascades or split protein constructs. Strengths and weaknesses of these genetic methods are discussed and some guidelines for verification of detected protein-protein interactions are provided.