Anti-ENG monoclonal antibody, clone NFN-330 [FITC]

Vascular development is determined by the sequential expression of specific genes, which can be studied by performing in situ hybridization assays in zebrafish during different developmental stages. To investigate the role of endoglin(eng) in vessel formation during the development of hereditary hemorrhagic telangiectasia (HHT), morpholino-mediated targeted knockdown of eng in zebrafish are used to study its temporal expression and associated functions. Here, whole-mount in situ RNA hybridization (WISH) is employed for the analysis of eng and its downstream genes in zebrafish embryos. Also, tube formation assays are performed in HHT patient-derived induced pluripotent stem cell-differentiated endothelial cells (iPSC-ECs; with eng mutations). A specific signal amplifying system using the whole amount In Situ Hybridization - WISH provides higher resolution and lower background results compared to traditional methods. To obtain a better signal, the post-fixation time is adjusted to 30 min after probe hybridization. Because fluorescence staining is not sensitive in zebrafish embryos, it is replaced with diaminobezidine (DAB) staining here. In this protocol, HHT patient-derived iPSC lines containing an eng mutation are differentiated into endothelial cells. After coating a plate with basement membrane matrix for 30 min at 37 degrees C, iPSC-ECs are seeded as a monolayer into wells and kept at 37 degrees C for 3 h. Then, the tube length and number of branches are calculated using microscopic images. Thus, with this improved WISH protocol, it is shown that reduced eng expression affects endothelial progenitor formation in zebrafish embryos. This is further confirmed by tube formation assays using iPSC-ECs derived from a patient with HHT. These assays confirm the role for eng in early vascular development.

Storm wave run-up causes beach erosion, wave overtopping, and street flooding. Extreme runup estimates may be improved, relative to predictions from general empirical formulae with default parameter values, by using historical storm waves and eroded profiles in numerical runup simulations. A climatology of storm wave run-up at Imperial Beach, California is developed using the numerical model SWASH, and over a decade of hindcast spectral waves and observed depth profiles. For use in a local flood warning system, the relationship between incident wave energy spectra E(f) and SWASH-modeled shoreline water levels is approximated with the numerically simple integrated power law approximation (IPA). Broad and multi-peaked E(f) are accommodated by characterizing wave forcing with frequency-weighted integrals of E(f). This integral approach improves runup estimates compared to the more commonly used bulk parameterization using deep water wave height H-0 and deep water wavelength L-0 Hunt (Trans Am Soc Civ Eng 126(4):542-570, 1961) and Stockdon et al. (Coast Eng 53(7):573-588, 2006. https://doi.org/10.1016/j.coast aleng.2005.12.005). Scaling of energy and frequency contributions in IPA, determined by searching parameter space for the best fit to SWASH, show an H0L0 scaling is near optimal. IPA performance is tested with LiDAR observations of storm run-up, which reached 2.5 m above the offshore water level, overtopped backshore riprap, and eroded the foreshore beach slope. Driven with estimates from a regional wave model and observed beta(f), the IPA reproduced observed run-up with < 30% error. However, errors in model physics, depth profile, and incoming wave predictions partially cancelled. IPA (or alternative empirical forms) can be calibrated (using SWASH or similar) for sites where historical waves and eroded bathymetry are available.