Advantages and Versatility of Fluorescence-Based Methodology to Characterize the Functionality of LDLR and Class Mutation Assignment

Familial hypercholesterolemia (FH) is a common autosomal codominant disease with a frequency of 1:500 individuals in its heterozygous form. The genetic basis of FH is most commonly mutations within the LDLR gene. Assessing the pathogenicity of LDLR variants is particularly important to give a patient a definitive diagnosis of FH. Current studies of LDLR activity ex vivo are based on the analysis of I-125-labeled lipoproteins (reference method) or fluorescent-labelled LDL. The main purpose of this study was to compare the effectiveness of these two methods to assess LDLR functionality in order to validate a functional assay to analyse LDLR mutations. LDLR activity of different variants has been studied by flow cytometry using FITC-labelled LDL and compared with studies performed previously with I-125-labeled lipoproteins. Flow cytometry results are in full agreement with the data obtained by the I-125 methodology. Additionally confocal microscopy allowed the assignment of different class mutation to the variants assayed. Use of fluorescence yielded similar results than I-125-labeled lipoproteins concerning LDLR activity determination, and also allows class mutation classification. The use of FITC-labelled LDL is easier in handling and disposal, cheaper than radioactivity and can be routinely performed by any group doing LDLR functional validations.

Basque nominalizations and the role of structural Case in the licensing of null arguments

[EN] This article investigates the question of the licensing of null arguments in the so-called pro-drop languages. By focusing on the licensing of null subjects in the different types of -T(Z)E nominalizations in Basque, it aims at defining in a precise way the crucial feature that makes pro-drop possible in a clause. The central claim is that what licenses subject-drop is the assignment of structural Case. That is, it is argued that a subject can be null if and only if it is assigned structural Case. Different aspects of T(Z)E nominalizations are also explored, which show that even if these clauses are similar in the surface, they can be syntactically very different and furthermore, that infinitive clauses marked with the same nominalizing morpheme can also have diverging structures.