The center of Rovinj is located on a peninsula, 500 m to the north of which is the marine station, ‘Acquario Berlinese

<p><b>Copyright information:</b></p><p>Taken from "Traditional and Modern Biomedical Prospecting: Part I—the History"</p><p>Evidence-based Complementary and Alternative Medicine 2004;1(1):71-82.</p><p>Published online Jan 2004</p><p>PMCID:PMC442115.</p><p>© Oxford University Press, 2004.</p>’ It was founded in 1891 (lithography from 1898); left. The institute was expanded in 1900 (photograph taken in 1902; postcard was sent on December 29, 1903 from the famous parasitologist Fritz Schaudinn to Stanislaus von Prowazek); right. The research vessel ‘Adria’ is shown (1898); lower left

Map of the Adriatic Sea as given in the ‘Pannoniae et Illyrici ve Teris Tabula’ by A

<p><b>Copyright information:</b></p><p>Taken from "Traditional and Modern Biomedical Prospecting: Part I—the History"</p><p>Evidence-based Complementary and Alternative Medicine 2004;1(1):71-82.</p><p>Published online Jan 2004</p><p>PMCID:PMC442115.</p><p>© Oxford University Press, 2004.</p> Ortelius 1590 (upper map). In the first edition of by S. Münster, the area around Rovinj (array) was not described (left) until 1582. In 1612, the name and the region was explicitly mentioned (right)

The <i>S. raphanus</i> putative carbonic anhydrase.

<p>(<b>A</b>) The sponge putative carbonic anhydrase (CA_SYCON) is aligned with the highly related sequences from the demosponge <i>S. domuncula</i>, the silicase (SIA_SUBDO; DD298191), and the carbonic anhydrases from the scleractinian <i>Acropora millepora</i> (CAr1_ACRMIL; ACJ64662.1), and the stony coral <i>Stylophora pistillata</i> (CAa_STYPI; ACA53457.1, EU159467.1), as well as with the human carbonic anhydrase 2 (CA II) (CAHB_HUMAN; O75493). The indicative sites/regions within the <i>Sycon</i> polypeptide are marked, the carbonic anhydrase alpha (vertebrate-like) group stretch (−CA−), including the His residues, functioning as Zn-binding sites, the hydrophobic parts (+hydb+), as well as the signal peptide (:signal:). Residues conserved (identical or similar) in all sequences are shown in white on black; those which share similarity to at least four residues are in black on grey. (<b>B</b>) Radial phylogenetic tree, including the mentioned sequences, together with human carbonic anhydrases of the following isoforms: I (CA-I) (CAH1_HUMAN; P00915); II (CA-II) (CAH2_HUMAN; P00918); III (CA-III) (CAH3_HUMAN; P07451); IV (CAIV_HUMAN; AAA35625.1); IV (CA-IV) (CAH4_HUMAN; P22748); VA (CAH5_HUMAN; P35218); VB (CA5B_HUMAN; CA5B_HUMAN); VI (CA-VI) (CAH6_HUMAN; P23280); VII (CA-VII) (CAH7_HUMAN; P43166); VIII (CA-VIII) (CAH8_HUMAN; P35219); IX (CA-IX) (CAH9_HUMAN; Q16790); 10 (CA-RP X) (CAHA_HUMAN; Q9NS85); XII (CA-XII) (CAHC_HUMAN; O43570); XIV (CA-XIV) (CAHE_HUMAN; Q9ULX7). In addition, the coral sequence from <i>Acropora millepora</i> (CAr2_ACRMIL; ACJ64663.1), as well as the ones from the sea anemone <i>Nematostella vectensis</i> (CAr_NEMVE; XP_001627923.1), the tunicate <i>Ciona intestinalis</i> (CA14_CIONA; XP_002123314.1); the lancelet <i>Branchiostoma floridae</i> (CAr_BRANFLO; XP_002601262.1), the shark <i>Squalus acanthias</i> (CA4_SQUAAC; AAZ03744.1); the fish <i>Oreochromis niloticus</i> (CA4_ORENI; XP_003456174.1), together with the insect enzyme from <i>D. melanogaster</i> (CAr_DROME; NP_572407.3) are included.</p

Expression levels of the OSTF (<i>SROSTFr</i>) in <i>S. raphanus</i> in dependence on the level of Ca²⁺ in the culture medium.

<p>Expression levels of the OSTF (<i>SROSTFr</i>) in <i>S. raphanus</i> in dependence on the level of Ca²⁺ in the culture medium.</p

Morphology of the bioglass preparation used.

<p>(<b>A</b> and <b>B</b>) SEM micrograph. The sintering temperature was 900°C.</p

SEM images of cell-free printed alginate/gelatin hydrogels.

<p>(<b>A</b>) Appearance of the basic alginate/gelatin hydrogel cylinders, containing neither silica nor biosilica nor polyP·Ca²⁺-complex. (<b>B</b>) Hydrogel containing 10 µg/ml of silicatein. (<b>C</b> and <b>D</b>) Printed basic alginate/gelatin hydrogel containing 50 µmoles/L ortho-silicate (>si<). (<b>E</b> and <b>F</b>) Cylinders of hydrogel supplemented with 50 µmoles/L biosilica (>bs<). (<b>G</b> and <b>H</b>) Alginate/gelatin hydrogel cylinders containing 100 µmoles/L polyP·Ca²⁺-complex (>polyP<). (<b>I</b> and <b>J</b>) Hydrogel containing both 50 µmoles/L biosilica (>bs<) and 100 µmoles/L polyP·Ca²⁺-complex.</p

The scheme depicts spicule formation via bio-inorganic self-organization.

<p>(<b>A</b>) The spicule (sp) synthesis starts intracellularly in sclerocytes (scl). The primordial spicules are associated with filaments (fi) which are assumed to participate in the extrusion of the growing spicule. This phase is dominated by the expression of silicatein that – at the later stage – is required for the formation of both the core and the shell cylinder of the siliceous mantel of the spicule. The newly formed silicatein molecules undergo fractal organization. (<b>B</b>) The primordial spicule is extruded and becomes associated in the extracellular space with sclerocytes (scl) which intracellularly form the silicasomes (sis). These organelles contain silicatein and silicate that are released into the extra-spicular space and cause bio-silica formation. (<b>C</b>) The growth of the spicule (sp) continues in two directions; axial elongation and appositional growth/thickening. The bio-silica formation is mediated by silicatein (sil) under the consumption of the substrate silicate (si). Growth of spicule is driven both longitudinally and (subsequently) radially along the cell protrusion. During this phase the cell extensions elongate by evagination. The core of the spicule mantel is formed by silicatein, existing in the axial canal, and the shell by silicatein layered onto the outer surface of the growing spicule. (<b>D</b>) Final completion of the size and form of the spicule. After termination the spicule disconnects from the sclerocyte (not shown in the scheme) and the hole is closed by bio-silica formation. The direction of cell movement is indicated with an arrow.</p

Results of the ELISA titration experiments with the phages Sycon-09 peptide, likely standing for the OSTF (Sycon-09(OSF)), and for the phage Sycon-23, indicative for the carbonic anhydrase (Sycon-23(CA)).

<p>The titer values (PFU/mL) obtained by using the antibodies against M13 revealed that the adsorption of Sycon-23(CA) and Sycon-09(OSTF) is much higher than the one read for the wild type phage M13.</p

Expression levels of the putative carbonic anhydrase (<i>SRCA</i>) gene in <i>S. raphanus</i>.

<p>The specimens were cultivated under standard condition (10 mM CaCl₂) or at Ca⁺⁺-depletion condition (1 mM CaCl₂). After incubation periods of 3 to 7 d the animals were collected, RNA prepared and the extent of gene expression was quantified by qRT-PCR, as described under “<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034617#s2" target="_blank">Material and Methods</a>”. Each data point represents the mRNA level of the respective expressed gene normalized to the amount of <i>β-tubulin</i> transcripts, as means ± SD (n = 6).</p

TEM images of the axial canal of spicules in primmorphs at different developmental stages.

<p>(<b>A</b>) The three major developmental phases during spicule formation: <u>phase I</u>: primordial spicule comprising a large axial canal (ac) which is surrounded by an organic cylinder enclosing vesicles (v). <u>phase II</u>: the spicule shows the siliceous mantel (si) surrounding the small axial canal (ac) devoid of a pronounced axial filament. <u>phase III</u>: such spicules have a small sized axial canal and a distinct axial filament (af); scl, sclerocyte. (<b>B</b>) Spicule in phase II. Membraneous structures can be resolved in the axial canal (ac) which is surrounded by the silica mantel (si). (<b>C</b>) Spicule between phases II and III showing in the axial canal a well developed axial filament (af) embedded in membranous structures, which is surrounded by the silica mantel (si) (<b>D</b>) Mature spicule with an axial filament (af) without any cellular structures. One surrounding sclerocyte is marked (scl). (<b>E</b>) Axial canal (ac), close to the apex of the spicule, comprising a homogenous granular material. (<b>F</b>) Spicule with an axial canal at phase II/III. The axial canal (ac) comprises a growing axial filament (af). (<b>G</b>) Intermediate spicule phase between II and III comprising an axial canal (ac) showing cellular structures with vesicles (v) and one axial filament (af).</p