Crassostrea patagonica (d'Orbigny, 1842) shell concentrations from the late Miocene Rio Negro province, NE Patagonia, Argentina

The Río Negro Formation (late Miocene-early Pliocene) mainly consists of continental deposits, but it contains a middle member of marine origin. It represents a transgressive-regressive sequence that can be seen at several outcrops along the N Patagonian coast. The taphonomical approach to the El Espigón marine deposits permits the identification of four main layers containing different kinds of skeletal accumulation, which mainly consist of oyster shells [Crassostrea patagonica (D'Orbigny, 1842)]. These concentrations display three different morphologies (pouches, pavements and bouquets) with a different taphonomic signature. These deposits were formed in shallow marine environments influenced by wave activity that produced valve concentrations of different entities. They contain several shell beds that represent event, composite, hiatal to lag skeletal concentrations. Traces of bioturbation in the sediment (Thalassinoides, Teichichnus) and bioerosion on the shells (Entobia, Gastrochaeonolites, Caulostrepsis), and encrusters (cirripeds, bryozoans), are also abundant in the outcrop and consititue common components of these Miocene materials. Layers 1 and 2 of the sequence were deposited in shoreface/foreshore environments at the beginning of a highstand systems tract, while layers 3 and 4 were deposited at the end, or at the beginning of a forced regression, in foreshore environments. A final erosional episode cut the top of the layer 4, which truncated the abundant bioturbaation developed there.

When cholesterol is not cholesterol: a note on the enzymatic determination of its concentration in model systems containing vegetable extracts

Background: Experimental evidences demonstrate that vegetable derived extracts inhibit cholesterol absorption in the gastrointestinal tract. To further explore the mechanisms behind, we modeled duodenal contents with several vegetable extracts. Results: By employing a widely used cholesterol quantification method based on a cholesterol oxidase-peroxidase coupled reaction we analyzed the effects on cholesterol partition. Evidenced interferences were analyzed by studying specific and unspecific inhibitors of cholesterol oxidase-peroxidase coupled reaction. Cholesterol was also quantified by LC/MS. We found a significant interference of diverse (cocoa and tea-derived) extracts over this method. The interference was strongly dependent on model matrix: while as in phosphate buffered saline, the development of unspecific fluorescence was inhibitable by catalase (but not by heat denaturation), suggesting vegetable extract derived H2O2 production, in bile-containing model systems, this interference also comprised cholesterol-oxidase inhibition. Several strategies, such as cholesterol standard addition and use of suitable blanks containing vegetable extracts were tested. When those failed, the use of a mass-spectrometry based chromatographic assay allowed quantification of cholesterol in models of duodenal contents in the presence of vegetable extracts. Conclusions: We propose that the use of cholesterol-oxidase and/or peroxidase based systems for cholesterol analyses in foodstuffs should be accurately monitored, as important interferences in all the components of the enzymatic chain were evident. The use of adequate controls, standard addition and finally, chromatographic analyses solve these issues.