Common Genetic Denominators for Ca++-Based Skeleton in Metazoa: Role of Osteoclast-Stimulating Factor and of Carbonic Anhydrase in a Calcareous Sponge

Calcium-based matrices serve predominantly as inorganic, hard skeletal systems in Metazoa from calcareous sponges [phylum Porifera; class Calcarea] to proto- and deuterostomian multicellular animals. The calcareous sponges form their skeletal elements, the spicules, from amorphous calcium carbonate (ACC). Treatment of spicules from Sycon raphanus with sodium hypochlorite (NaOCl) results in the disintegration of the ACC in those skeletal elements. Until now a distinct protein/enzyme involved in ACC metabolism could not been identified in those animals. We applied the technique of phage display combinatorial libraries to identify oligopeptides that bind to NaOCl-treated spicules: those oligopeptides allowed us to detect proteins that bind to those spicules. Two molecules have been identified, the (putative) enzyme carbonic anhydrase and the (putative) osteoclast-stimulating factor (OSTF), that are involved in the catabolism of ACC. The complete cDNAs were isolated and the recombinant proteins were prepared to raise antibodies. In turn, immunofluorescence staining of tissue slices and qPCR analyses have been performed. The data show that sponges, cultivated under standard condition (10 mM CaCl2) show low levels of transcripts/proteins for carbonic anhydrase or OSTF, compared to those animals that had been cultivated under Ca2+-depletion condition (1 mM CaCl2). Our data identify with the carbonic anhydrase and the OSTF the first two molecules which remain conserved in cells, potentially involved in Ca-based skeletal dissolution, from sponges (sclerocytes) to human (osteoclast)

Search for pair production of scalar leptoquarks decaying into first- or second-generation leptons and top quarks in proton–proton collisions at s = 13 TeV with the ATLAS detector

Abstract: A search for pair production of scalar leptoquarks, each decaying into either an electron or a muon and a top quark, is presented. This is the first leptoquark search using ATLAS data to investigate top-philic cross-generational couplings that could provide explanations for recently observed anomalies in B meson decays. This analysis targets high leptoquark masses which cause the decay products of each resultant top quark to be contained within a single high-pT large-radius jet. The full Run 2 dataset is exploited, consisting of 139fb-1 of data collected from proton–proton collisions at s=13TeV from 2015 to 2018 with the ATLAS detector at the CERN Large Hadron Collider. In the absence of any significant deviation from the background expectation, lower limits on the leptoquark masses are set at 1480GeV and 1470GeV for the electron and muon channel, respectively