Annulatascus aquatorba sp. nov., a lignicolous freshwater ascomycete from Sirindhorn Peat Swamp Forest, Narathiwat, Thailand

As part of a long term study of fungi colonizing submerged wood in freshwater streams a new Annulatascus species, A. aquatorba, is described and illustrated from Erythrophleum teysmannii test blocks from Sirindhorn Peat Swamp Forest, southern Thailand. It differs from other species in the genus in ascospore measurements, thickness of the cell wall, 1-3-septate, fusoid to lunate shape, with central brown cells and subhyaline end cells and without a mucilaginous sheath. Asci are cylindrical, pedicellate, with a distinct, wedge-shaped and non-amyloid apical ring. Phylogenetic relationships of this species, based on the combined partial 18S and 28S rDNA, place it in the same clade as A. velatisporus (type species), A. hongkongensis and A. nilensis.     

Inhibition of Quorum Sensing Mediated Virulence Factors Production in Urinary Pathogen Serratia marcescens PS1 by Marine Sponges

The focal intent of this study was to find out an alternative strategy for the antibiotic usage against bacterial infections. The quorum sensing inhibitory (QSI) activity of marine sponges collected from Palk Bay, India was evaluated against acyl homoserine lactone (AHL) mediated violacein production in Chromobacterium violaceum (ATCC 12472), CV026 and virulence gene expressions in clinical isolate Serratia marcescens PS1. Out of 29 marine sponges tested, the methanol extracts of Aphrocallistes bocagei (TS 8), Haliclona (Gellius) megastoma (TS 25) and Clathria atrasanguinea (TS 27) inhibited the AHL mediated violacein production in C. violaceum (ATCC 12472) and CV026. Further, these sponge extracts inhibited the AHL dependent prodigiosin pigment, virulence enzymes such as protease, hemolysin production and biofilm formation in S. marcescens PS1. However, these sponge extracts were not inhibitory to bacterial growth, which reveals the fact that the QSI activity of these extracts was not related to static or killing effects on bacteria. Based on the obtained results, it is envisaged that the marine sponges could pave the way to prevent quorum sensing (QS) mediated bacterial infections.     

The Littorina sequence database (LSD)--an online resource for genomic data

We present an interactive, searchable expressed sequence tag database for the periwinkle snail Littorina saxatilis, an upcoming model species in evolutionary biology. The database is the result of a hybrid assembly between Sanger and 454 sequences, 1290 and 147,491 sequences respectively. Normalized and non-normalized cDNA was obtained from different ecotypes of L. saxatilis collected in the UK and Sweden. The Littorina sequence database (LSD) contains 26,537 different contigs, of which 2453 showed similarity with annotated proteins in UniProt. Querying the LSD permits the selection of the taxonomic origin of blast hits for each contig, and the search can be restricted to particular taxonomic groups. The database allows access to UniProt annotations, blast output, protein family domains (PFAM) and Gene Ontology. The database will allow users to search for genetic markers and identifying candidate genes or genes for expression analyses. It is open for additional deposition of sequence information for L. saxatilis and other species of the genus Littorina. The LSD is available at http://mbio-serv2.mbioekol.lu.se/Littorina/.     

In vitro activation of the IkappaB kinase complex by human T-cell leukemia virus type-1 Tax

Human T-cell leukemia virus type-I expresses Tax, a 40-kDa oncoprotein that activates IkappaB kinase (IKK), resulting in constitutive activation of NFkappaB. Herein, we have developed an in vitro signaling assay to analyze IKK complex activation by recombinant Tax. Using this assay in combination with reporter assays, we demonstrate that Tax-mediated activation of IKK is independent of phosphatases. We show that sustained activation of the Tax-mediated activation of the NFkappaB pathway is dependent on an intact Hsp90-IKK complex. By acetylating and thereby preventing activation of the IKK complex by the Yersinia effector YopJ, we demonstrate that Tax-mediated activation of the IKK complex requires a phosphorylation step. Our characterization of an in vitro signaling assay system for the mechanism of Tax-mediated activation of the IKK complex with a variety of mutants and inhibitors results in a working model for the biochemical mechanism of Tax-induced activation.     

Identifying a Kinase Network Regulating FGF14:Nav1.6 Complex Assembly Using Split-Luciferase Complementation

Kinases play fundamental roles in the brain. Through complex signaling pathways, kinases regulate the strength of protein:protein interactions (PPI) influencing cell cycle, signal transduction, and electrical activity of neurons. Changes induced by kinases on neuronal excitability, synaptic plasticity and brain connectivity are linked to complex brain disorders, but the molecular mechanisms underlying these cellular events remain for the most part elusive. To further our understanding of brain disease, new methods for rapidly surveying kinase pathways in the cellular context are needed. The bioluminescence-based luciferase complementation assay (LCA) is a powerful, versatile toolkit for the exploration of PPI. LCA relies on the complementation of two firefly luciferase protein fragments that are functionally reconstituted into the full luciferase enzyme by two interacting binding partners. Here, we applied LCA in live cells to assay 12 kinase pathways as regulators of the PPI complex formed by the voltage-gated sodium channel, Nav1.6, a transmembrane ion channel that elicits the action potential in neurons and mediates synaptic transmission, and its multivalent accessory protein, the fibroblast growth factor 14 (FGF14). Through extensive dose-dependent validations of structurally-diverse kinase inhibitors and hierarchical clustering, we identified the PI3K/Akt pathway, the cell-cycle regulator Wee1 kinase, and protein kinase C (PKC) as prospective regulatory nodes of neuronal excitability through modulation of the FGF14:Nav1.6 complex. Ingenuity Pathway Analysis shows convergence of these pathways on glycogen synthase kinase 3 (GSK3) and functional assays demonstrate that inhibition of GSK3 impairs excitability of hippocampal neurons. This combined approach provides a versatile toolkit for rapidly surveying PPI signaling, allowing the discovery of new modular pathways centered on GSK3 that might be the basis for functional alterations between the normal and diseased brain.     

Study of β-III tubulin expression in human eye tissues during prenatal development

The expression of β-III tubulin, a marker protein of early neuronal cells, was studied by molecular genetic and immunochemical techniques. The study was performed with the eyes of human fetuses on weeks 8.5 to 27–28 of intrauterine development. Expression of β-III tubulin was detected immunochemically in the retina and lens fibers of fetuses at stages of 8.5 to 22–23 weeks. By means of PCR, a strong expression of the β-III tubulin gene was revealed in the retina of 9.5-week fetuses. Its level remained high until week 18, became slightly lower by week 24, and decreased to zero by week 27–28. The expression of this gene was also revealed in the lens of 9.5-week fetuses. Its level at stages of 15 to 24 weeks was very low, and no expression could be detected in 27-to 28-week fetuses. The results of PCR analysis were consistent with immunochemical data.     

Ploidies of cardiomyocytes in human myocardial hypertrophy]

DNA cytophotometry has been performed in ventricular cardiomyocytes of hypertrophic human hearts. In the cases of hypertrophy in adults (generalized atherosclerosis, postinfarct scars), polyploidy expression did not exceed the limits of normal variability developed during childhood. In the cases of hypertrophy caused by congenital heart defects, high polyploidy has been revealed (the mean level 20c and more, where c is haploid DNA content), which considerably exceeded the upper limit of normal variability (approximately 10c). Our hypothesis has confirmed that heart hypertrophy in adults proceeds in conditions of stable genome rather than due to redundant polyploidization of the ventricular myocytes. The same idea assumes enhanced polyploidization of the myocytes in childhood in humans with congenital heart diseases.     

A molecular phylogeny of the north‐east Atlantic species of the genus Idotea (Isopoda) with focus on the Baltic Sea

Today, the marine isopods of the genus Idotea Fabricius, 1798 consist of 26 accepted species. Most species can be found in the Northern Hemisphere. While some species have a cosmopolitan distribution, others are endemic to a few coastlines. In the Baltic Sea, Idotea species have a key role as important meso‐grazers. Only three species can be found in this brackish environment, I. balthica, I. chelipes and I. granulosa, while nine species are described from the whole European coast. The goal of the present study was to reconstruct the phylogeny of the genus Idotea and to test whether the Baltic low‐salinity tolerant species represent a single lineage within the genus. The phylogeny of north‐east Atlantic Idotea species was investigated based on a fragment of the nuclear gene 28S and the mitochondrial gene COI for eight Idotea species. The phylogenetic reconstructions showed that the genus Idotea might not be monophyletic. Neither I. linearis nor I. urotoma did belong to the separated and well‐supported Idotea clade of I. balthica, I. chelipes, I. emarginata, I. granulosa, I. metallica and I. pelagica. The three Idotea species found in the Baltic belonged to different lineages, with estimated COI‐based divergence time older than 7 Myr. This suggests that the tolerance to low salinity has likely evolved in the genus Idotea more than once.     

Individual patterns of tonal (whistling) signals of bottlenose dolphins (Tursiops truncates) kept in relative isolation

The results of study of the subaqueous acoustic signaling of bottlenose dolphins (Tursiopsis truncatus) in a dolphinarium are shown. Individual patterns of whistling signals of four individuals were described and analyzed. To determine which animal produced the sound, the method of ??relative isolation?? was used. It was found that the pattern included whistles specific for a particular individual (??signature whistles??). The percentage of these whistles in the patterns varied considerably among individuals. The signatures had variable frequency and duration. The phenomenon of ??imitation?? was typical for animals at higher positions in the hierarchy: generating ??signatures?? specific for other individuals. Besides the signatures, variable signals and whistles with simple frequency contours were found in the patterns.     

Morphometric characteristics of hepatocyte mitochondria during internal administration of boron-containing water

The changes of hepatic mitochondrial cells were studied in white male rats during internal application of boron-containing waters, having 250 mg/l concentration of boron, but different ionic-salt basis. Mitochondrial hepatocytes and hepatic boron content relationships were established. It was shown that during boron accumulation in liver (15 applications) decreasing of mitochondrial size occurred, independently, of the type of boron-containing waters. During stable boron level and particularly during its excretion from organism (24 applications, 6 days aftereffect), increase of size of mitochondria and their ellipticity, change of configuration, that depend on the type of applied waters was shown.