Deep Mining of Oxysterols and Cholestenoic Acids in Human Plasma and Cerebrospinal Fluid: Quantification using Isotope Dilution Mass Spectrometry

Both plasma and cerebrospinal fluid (CSF) are rich in cholesterol and its metabolites. Here we describe in detail a methodology for the identification and quantification of multiple sterols including oxysterols and sterol-acids found in these fluids. The method is translatable to any laboratory with access to liquid chromatography – tandem mass spectrometry. The method exploits isotope-dilution mass spectrometry for absolute quantification of target metabolites. The method is applicable for semi-quantification of other sterols for which isotope labelled surrogates are not available and approximate quantification of partially identified sterols. Values are reported for non-esterified sterols in the absence of saponification and total sterols following saponification. In this way absolute quantification data is reported for 17 sterols in the NIST SRM 1950 plasma along with semi-quantitative data for 8 additional sterols and approximate quantification for one further sterol. In a pooled (CSF) sample used for internal quality control, absolute quantification was performed on 10 sterols, semi-quantification on 9 sterols and approximate quantification on a further three partially identified sterols. The value of the method is illustrated by confirming the sterol phenotype of a patient suffering from ACOX2 deficiency, a rare disorder of bile acid biosynthesis, and in a plasma sample from a patient suffering from cerebrotendinous xanthomatosis, where cholesterol 27-hydroxylase is deficient

Brachiopod palaeobiogeography in the western Tethys during the Early Jurassic diversity maximum: introduction of a Pontic Province

The biogeography of Pliensbachian (Early Jurassic) brachiopods in western Tethys is investigated using complementary multivariate tools including metric and non-metric ordination, additive cluster and bootstrapped spanning network analyses, as well as one-way analysis of similarity and similarity percentage analysis. All analyses were conducted using the Dice and Simpson similarity indices for presence/absence data on occurrence datasets involving 24 assemblages homogenized at the species level, including (403 taxa) or excluding (210 taxa) species found in only one assemblage. The analyses present a highly consistent biogeographical picture involving three main clusters: the Euro-Boreal, Mediterranean and Pontic biochores. The brachiopod species typical of the newly defined Pontic biochore are illustrated. The three assemblages from the Atlas area are interpreted as a fourth biochore, compositionally intermediate between those of the Euro-Boreal and Mediterranean. The Mediterranean biochore can be further divided into an intra- and a peri-Mediterranean group. These five, palaeogeographically well-constrained biochores show moderate to high degrees of species endemicity, ranging from 20% (Atlas) to 58% (Euro-Boreal). Based on available evidence, and after a reasonable cutback of the customary scale of ranks, the following biogeographical categories and names are suggested for the western Tethyan Pliensbachian brachiopod biochores: Euro-Boreal Province, Mediterranean Province (including an intra-Mediterranean and a peri-Mediterranean Subprovince), Pontic Province and Atlas Subprovince. In addition, a still poorly documented brachiopod biochore occurs on the Gondwana margin as a possible precursor of the extensive Middle to Late Jurassic Ethiopian Province