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The pathophysiology of AKI during tumor lysis syndrome (TLS) is not fully understood. We aimed to decipher crystal-dependent and crystal-independent mechanisms of TLS-induced AKI. Analyzing urine and blood from patients with TLS provided data on crystal-independent mechanisms of the pathogenesis of AKI during TLS. We also explored mechanisms of TLS-induced AKI in vitroand in vivoin a murine model of TLS (syngeneic mice with acute myeloid leukemia receiving chemotherapy). We found that extracellular histones released in huge amounts during TLS profoundly alter the endothelium. Nonanticoagulant heparin mitigated AKI in this model.

Chemical investigation of the Mediterranean calcareous sponge Clathrina clathrusled to the isolation of large amounts of a new 2-aminoimidazole alkaloid, named clathridimine (1), along with the known clathridine (2) and its zinc complex (3). The structure of the new metabolite was assigned by detailed spectroscopic analysis. Clathridimine (1) displayed selective anti-Escherichia coliand anti-Candida albicansactivities. Clathridine (2) showed only anti-Candida albicansactivity, and its zinc complex (3) exhibited selective anti-Staphylococcus aureusactivity. The isolation of analogues of 2-amino-imidazole derivatives from several Leucettaspecies from various sites in the Pacific Ocean and the Red Sea raises the question of their biosynthetic origin. Microscopic studies revealed abundant extracellular bacteria located in the mesohyl of the sponge, with two predominant morphotypes including spiral bacteria and long, narrow bacilli. Chemical analysis with HPLC/UV/ELSD profiles of sponge cells separated from bacteria by differential centrifugation and trypsinization of the sponge cell surface revealed that clathridine (2) was localized in the sponge cells.

Microcin E492 (MccE492) is an antibacterial peptide naturally secreted by Klebsiella pneumoniaeRYC492. Initially described as an 84-residue unmodified peptide, it was also recently isolated in a posttranslationally modified form, MccE492m. The production of MccE492m is dependent on the synthesis of enterobactin and the mceABCDEFGHIJgene cluster. The posttranslational modification was characterized as a trimer of N-(2,3-dihydroxybenzoyl)-l-serine (DHBS) linked to the Ser84-carboxylate viaa β-d-glucose moiety. MccE492m was shown to bind ferric ions through the trimer of DHBS. This is the first example of a novel type of antibacterial peptide termed siderophore-peptide. Recognition of MccE492m, but also of the unmodified MccE492, was shown to be mediated by the catecholate siderophore receptors FepA, Cir and Fiu at the outer membrane of E. coli. The siderophore-type modification was shown to be responsible for a significant enhancement of the microcin antibacterial activity. Therefore, we propose that MccE492 and MccE492m use iron-siderophore receptors for uptake into the target bacteria and that improvement of MccE492 antimicrobial activity upon modification results from an increase in the microcin/receptor affinity.

We previously described that mouse oocytes and preimplantation embryos express the two subunits of interferon-gamma receptor. We now report that, despite the presence of STAT1 (signal transducer and activator of transcription 1) at both the mRNA and protein levels, interferon γ (IFNγ) as well as IFNα are unable to trigger massive nuclear translocation of STAT1 in these cells, even at high cytokine concentrations. Conversely, nuclear accumulation of STAT1 was readily observed in murine L929 somatic cells under the same conditions. However, in the absence of any stimulation, both tyrosine (Y701p) and serine (S727p) phosphorylated forms of STAT1 were already detected in the nuclei of oocytes and early embryos. Phosphorylated STAT1 appeared concentrated in large nuclear dots, which were identified by indirect immunofluorescence and electron microscopy as clusters of interchromatin granules (IGCs or speckles). A similar distribution was also observed for the serine (S727p) phosphorylated form of STAT3 as well as for tyrosine (Y689p) phosphorylated STAT2. Western blot analysis confirmed that STAT factors present in mouse oocytes are predominantly phosphorylated. In parallel, we showed that the transcription of two IFNγ-target genes, namely interferon regulatory factor-1 (IRF-1) and suppressor of cytokine signaling-1 (SOCS-1) is indeed increased in two-cell embryos in response to IFNγ. Altogether, our results suggest that, despite the lack of massive nuclear accumulation of STAT1 in response to exogenous IFNs and the permanent presence of phosphorylated STATs in the nucleus, JAK/STAT pathways are functional during early development.

Chaetognaths (arrow worms) are abundant hermaphrodite marine organisms whose phylogenetic position amongst protostomes and deuterostomes is still debated. Ancient histological observations dating from a century ago described the presence in eggs of a large granule, presumed to be a germ plasm, and its probable inheritance in four primary germ cells (PGCs). Using videomicroscopy, electron microscopy and immunocytochemistry (labelling with anti-Vasa antibodies) we have followed the cycle of aggregation and dispersion of germ plasm and nuage material in eggs, embryos, PGCs and oocytes in several species of benthic (Spadella) and planctonic (Sagitta) chaetognaths. In these animals, germ cells and gametes can be observed in vivo throughout the 1-2 month life cycle. After describing internal fertilization in live animals we show that the single large (15 μm diameter) germ granule forms by a spiralling aggregation movement of small germ islands situated in the vegetal cortex at the time of first mitosis. We also demonstrate that the granule forms autonomously in unfertilized activated eggs or fertilized egg fragments. Once formed, the germ granule first associates with the cleavage furrow and is segregated into one of the first two blastomeres. The germ granule is then translocated from the cortex to the mitotic spindle during 3rd cleavage and remains in the single most-vegetal blastomere until the 32-cell stage. At the 64-cell stage the germ granule is partitioned as nuage material into two founder PGCs and further partitioned into four PGCs situated at the tip of the archenteron during gastrulation. These four PGCs migrate without dividing to reach the transverse septum, then proliferate and differentiate into oocytes and spermatocytes of two ovaries and two testes. We noted that germ plasm and nuage material were associated with mitochondria, the nucleus, the spindle and the centrosome during some stages of development and differentiation of the germ line. Finally, we demonstrate that a Vasa-like protein is present in the germ granule, in PGCs and in the electron-dense material associated with the germinal vesicle of oocytes. These features stress the conservation of cellular and molecular mechanisms involved in germ cell determination. Movies available on-line

Many eggs undergo reorganizations that localize determinants specifying the developmental axes and the differentiation of various cell types. In ascidians, fertilization triggers spectacular reorganizations that result in the formation and localization of distinct cytoplasmic domains that are inherited by early blastomeres that develop autonomously. By applying various imaging techniques to the transparent eggs of Phallusia mammillata, we now define 9 events and phases in the reorganization of the surface, cortex and the cytoplasm between fertilization and first cleavage. We show that two of the domains that preexist in the egg (the ER-rich cortical domain and the mitochondria-rich subcortical myoplasm) are localized successively by a microfilament-driven cortical contraction, a microtubule-driven migration and rotation of the sperm aster with respect to the cortex, and finally, a novel microfilament-dependant relaxation of the vegetal cortex. The phases of reorganization we have observed can best be explained in terms of cell cycle-regulated phases of coupling, uncoupling and recoupling of the motions of cortical and subcortical layers (ER-rich cortical domain and mitochondria-rich domain) with respect to the surface of the zygote. At the end of the meiotic cell cycle we can distinguish up to 5 cortical and cytoplasmic domains (including two novel ones; the vegetal body and a yolk-rich domain) layered against the vegetal cortex. We have also analyzed how the myoplasm is partitioned into distinct blastomeres at the 32-cell stage and the effects on development of the ablation of precisely located small fragments. On the basis of our observations and of the ablation/ transplantation experiments done in the zygotes of Phallusia and several other ascidians, we suggest that the determinants for unequal cleavage, gastrulation and for the differentiation of muscle and endoderm cells may reside in 4 distinct cortical and cytoplasmic domains localized in the egg between fertilization and cleavage.