Pomegranate liquor preparation and analysis

The pomegranate (Punica granatum L.) liquor has been produced for several centuries in the south of Portugal, mainly in the mountain areas. The “Assaria” variety is the preferred cultivar due to its organoleptic properties and high arils to peel ratio. Wild pomegranates are also widely distributed but, despite the health benefits that have been associated to the fruits, they continue to be unappreciated for consumption. Liquor preparation is a very good alternative for wild pomegranate fruits. We prepared pomegranate liquors by following a maceration procedure using the arils or juice of Assaria and wild pomegranate fruits. Strawberry tree (Arbutus unedo L.) fruit spirits were used to prepare the liquors. At the end of the maceration time 5 day as minimum sugar syrup was added. The maturation period was three months or longer. The obtained liquors showed a very attractive pink colour. The colour and the total polyphenol, as well as the anthocyanin and ellagitannin profiles, were measured at the end of the maceration and maturation times. Wild pomegranates gave rise liquors with more intense pink colour and higher polyphenol contents than the prepared using Assaria fruits. The anthocyanin and ellagitannin profiles also indicated higher contents of polyphenols for liquors prepared using wild pomegranate fruits. When juice is used instead of complete arils during the maceration period punicalin is not present and the consequently total polyphenols is low. The main anthocyanins identified in the liquors were delphinidin-3,5-diglucoside, cyaniding-3,5-diglucoside, delphinidin–3-glucoside, cyaniding–3-glucoside, pelargonidin–3–glucoside; the main ellagitannins were punicalagin and punicalin.

Evolution of matrix and bone gamma-carboxyglutamic acid proteins in vertebrates

The evolution of calcified tissues is a defining feature in vertebrate evolution. Investigating the evolution of proteins involved in tissue calcification should help elucidate how calcified tissues have evolved. The purpose of this study was to collect and compare sequences of matrix and bone γ-carboxyglutamic acid proteins (MGP and BGP, respectively) to identify common features and determine the evolutionary relationship between MGP and BGP. Thirteen cDNAs and genes were cloned using standard methods or reconstructed through the use of comparative genomics and data mining. These sequences were compared with available annotated sequences (a total of 48 complete or nearly complete sequences, 28 BGPs and 20 MGPs) have been identified across 32 different species (representing most classes of vertebrates), and evolutionarily conserved features in both MGP and BGP were analyzed using bioinformatic tools and the Tree-Puzzle software. We propose that: 1) MGP and BGP genes originated from two genome duplications that occurred around 500 and 400 million years ago before jawless and jawed fish evolved, respectively; 2) MGP appeared first concomitantly with the emergence of cartilaginous structures, and BGP appeared thereafter along with bony structures; and 3) BGP derives from MGP. We also propose a highly specific pattern definition for the Gla domain of BGP and MGP. Previous Section Next Section BGP1 (bone Gla protein or osteocalcin) and MGP (matrix Gla protein) belong to the growing family of vitamin K-dependent (VKD) proteins, the members of which are involved in a broad range of biological functions such as skeletogenesis and bone maintenance (BGP and MGP), hemostasis (prothrombin, clotting factors VII, IX, and X, and proteins C, S, and Z), growth control (gas6), and potentially signal transduction (proline-rich Gla proteins 1 and 2). VKD proteins are characterized by the presence of several Gla residues resulting from the post-translational vitamin K-dependent γ-carboxylation of specific glutamates, through which they can bind to calcium-containing mineral such as hydroxyapatite. To date, VKD proteins have only been clearly identified in vertebrates (1) although the presence of a γ-glutamyl carboxylase has been reported in the fruit fly Drosophila melanogaster (2) and in marine snails belonging to the genus Conus (3). Gla residues have also been found in neuropeptides from Conus venoms (4), suggesting a wider prevalence of γ-carboxylation.