The dephosphorylated form of the anaphase-promoting complex protein Cdc27/Apc3 concentrates on kinetochores and chromosome arms in mitosis

Cell cycle regulated protein ubiquitination and degradation within subcellular domains may be essential for the normal progression of mitosis. Cdc27 is a conserved component of an essential M-phase ubiquitin-protein ligase called the anaphase-promoting complex/cyclosome. We examined the subcellular distribution of Cdc27 in greater detail in mammalian cells and found Cdc27 concentrated at spindle poles and on spindle microtubules as previously described, but also found Cdc27 at kinetochores and along chromosome arms. This localization was not dependent on intact microtubules. While the great majority of Cdc27 protein in M phase cells is highly phosphorylated, only the dephosphorylated form of Cdc27 was found associated with isolated chromosomes. Kinases that also associate with isolated chromosomes catalyzed the in vitro phosphorylation of the chromosome-associated Cdc27. Microinjection of anti-Cdc27 antibody into cells causes arrest at metaphase. Microinjection of cells with anti-Mad2 antibody normally induces premature anaphase onset resulting in catastrophic nondisjunction of the chromosomes. However, coinjection of anti-Cdc27 antibody with anti-Mad2 antibody resulted in metaphase arrest. The association of dephosphorylated APC/C components with mitotic chromosomes suggests mechanisms by which the spindle checkpoint may regulate APC/C activity at mitosis.     

Ribosomal Protein Genes RPS10 and RPS26 Are Commonly Mutated in Diamond-Blackfan Anemia

Diamond-Blackfan anemia (DBA), an inherited bone marrow failure syndrome characterized by anemia that usually presents before the first birthday or in early childhood, is associated with birth defects and an increased risk of cancer. Although anemia is the most prominent feature of DBA, the disease is also characterized by growth retardation and congenital malformations, in particular craniofacial, upper limb, heart, and urinary system defects that are present in ∼30%–50% of patients. DBA has been associated with mutations in seven ribosomal protein (RP) genes, RPS19, RPS24, RPS17, RPL35A, RPL5, RPL11, and RPS7, in about 43% of patients. To continue our large-scale screen of RP genes in a DBA population, we sequenced 35 ribosomal protein genes, RPL15, RPL24, RPL29, RPL32, RPL34, RPL9, RPL37, RPS14, RPS23, RPL10A, RPS10, RPS12, RPS18, RPL30, RPS20, RPL12, RPL7A, RPS6, RPL27A, RPLP2, RPS25, RPS3, RPL41, RPL6, RPLP0, RPS26, RPL21, RPL36AL, RPS29, RPL4, RPLP1, RPL13, RPS15A, RPS2, and RPL38, in our DBA patient cohort of 117 probands. We identified three distinct mutations of RPS10 in five probands and nine distinct mutations of RPS26 in 12 probands. Pre-rRNA analysis in lymphoblastoid cells from patients bearing mutations in RPS10 and RPS26 showed elevated levels of 18S-E pre-rRNA. This accumulation is consistent with the phenotype observed in HeLa cells after knockdown of RPS10 or RPS26 expression with siRNAs, which indicates that mutations in the RPS10 and RPS26 genes in DBA patients affect the function of the proteins in rRNA processing.     

Caspase-1-driven neutrophil pyroptosis and its role in host susceptibility to Pseudomonas aeruginosa

Multiple regulated neutrophil cell death programs contribute to host defense against infections. However, despite expressing all necessary inflammasome components, neutrophils are thought to be generally defective in Caspase-1-dependent pyroptosis. By screening different bacterial species, we found that several Pseudomonas aeruginosa (P. aeruginosa) strains trigger Caspase-1-dependent pyroptosis in human and murine neutrophils. Notably, deletion of Exotoxins U or S in P. aeruginosa enhanced neutrophil death to Caspase-1-dependent pyroptosis, suggesting that these exotoxins interfere with this pathway. Mechanistically, P. aeruginosa Flagellin activates the NLRC4 inflammasome, which supports Caspase-1-driven interleukin (IL)-1β secretion and Gasdermin D (GSDMD)-dependent neutrophil pyroptosis. Furthermore, P. aeruginosa-induced GSDMD activation triggers Calcium-dependent and Peptidyl Arginine Deaminase-4-driven histone citrullination and translocation of neutrophil DNA into the cell cytosol without inducing extracellular Neutrophil Extracellular Traps. Finally, we show that neutrophil Caspase-1 contributes to IL-1β production and susceptibility to pyroptosis-inducing P. aeruginosa strains in vivo. Overall, we demonstrate that neutrophils are not universally resistant for Caspase-1-dependent pyroptosis.     

Characterization of the retinoic acid receptor genes raraa, rarab and rarg during zebrafish development

Retinoic acid signaling is important for patterning the central nervous system, paired appendages, digestive tract, and other organs. To begin to investigate retinoic acid signaling in zebrafish, we determined orthologies between zebrafish and tetrapod retinoic acid receptors (Rars) and examined the expression patterns of rar genes during embryonic development. Analysis of phylogenies and conserved syntenies showed that the three cloned zebrafish rar genes include raraa and rarab, which are co-orthologs of tetrapod Rara, and rarg, which is the zebrafish ortholog of tetrapod Rarg. We did not, however, find an ortholog of Rarb. RNA in situ hybridization experiments showed that rarab and rarg, are maternally expressed. Zygotic expression of raraa occurs predominantly in the hindbrain, lateral mesoderm, and tailbud. Zygotic expression of rarab largely overlaps that of raraa, except that in later stages rarab is expressed more broadly in the brain and in the pectoral fin bud and pharyngeal arches. Zygotic expression of zebrafish rarg also overlaps the other two genes, but it is expressed more strongly in the posterior hindbrain beginning in late somitogenesis as well as in neural crest cells in the pharyngeal arches. Thus, these three genes have largely overlapping expression patterns and a few gene-specific expression domains. Knowledge of these expression patterns will guide the interpretation of the roles these genes play in development.     

Species–area relationships in the Andaman and Nicobar Islands emerge because rarer species are disproportionately favored on larger islands

The island species–area relationship (ISAR) describes how the number of species increases with increasing size of an island (or island‐like habitat), and is of fundamental importance in island biogeography and conservation. Here, we use a framework based on individual‐based rarefaction to infer whether ISARs result from passive sampling, or whether some processes are acting beyond sampling (e.g., disproportionate effects and/or habitat heterogeneity). Using data on total and relative abundances of four taxa (birds, butterflies, amphibians, and reptiles) from multiple islands in the Andaman and Nicobar archipelago, we examine how different metrics of biodiversity (total species richness, rarefied species richness, and abundance‐weighted effective numbers of species emphasizing common species) vary with island area. Total species richness increased for all taxa, as did rarefied species richness controlling for a given sampling effort. This indicates that the ISAR did not result because of passive sampling, but that instead, some species were disproportionately favored on larger islands. For birds, frogs, and lizards, this disproportionate effect was only associated with species that were rarer in the samples, but for butterflies, both more common and rarer species were affected. Furthermore, for the two taxa for which we had plot‐level data (reptiles and amphibians), within‐island β‐diversity did not increase with island size, suggesting that within‐island compositional effects were unlikely to be driving these ISARs. Overall, our results indicate that the ISARs of these taxa are most likely driven by disproportionate effects, that is, where larger islands are important sources of biodiversity beyond a simple sampling expectation, especially through their influence on rarer species, thus emphasizing their role in the preservation and conservation of species.     

Pollinator dependence but no pollen limitation for eight plants occurring north of the Arctic Circle

Effective interactions between plants and pollinators are essential for the reproduction of plant species. Pollinator exclusion experiments and pollen supplementation experiments quantify the degree to which plants depend on animal pollinators and the degree to which plant reproduction is pollen limited. Pollen supplementation experiments have been conducted across the globe, but are rare in high latitude regions. To fill this knowledge gap, we experimentally investigated the dependence on animal pollinators and magnitude of pollen limitation in eight plant species north of the Arctic Circle in Lapland, Finland. Our findings show that all plant species were pollinator dependent, but not pollen limited. We discuss several mechanisms that might buffer our focal plants from pollen limitation, including plant and pollinator generalization, and attractive plant traits. Our results demonstrate that many plant species north of the Arctic Circle are currently receiving adequate pollinator service and provide a baseline for future comparisons of pollinator dependence and pollen limitation in the Arctic across space and time.     

Characterization of retinoid-X receptor genes rxra, rxrba, rxrbb and rxrg during zebrafish development

During development of vertebrate embryos, retinoic acid plays a variety of roles that are mediated by binding to retinoic acid receptors (Rars) and their heterodimerization partners, the retinoid receptors (Rxrs). Here, we characterize the expression patterns of four zebrafish rxr genes during development and provide an analysis of the phylogenetic relationships between zebrafish and tetrapod Rxr genes based on sequence similarities and conserved syntenies. This analysis prompted the renaming of several of the zebrafish rxr genes to match their tetrapod orthologs. Understanding phylogenetic relationships among Rxr genes and their expression patterns during development provides a foundation for future studies of Rxr functions.     

Increasing zoo’s conservation potential through understanding barriers to holding globally threatened amphibians

The global amphibian crisis and current un-mitigatable threats make ex situ programmes a crucial complementary action for the conservation of many amphibians. Zoos and aquariums are some of the most important and influential groups of institutions to undertake this yet the proportion of globally threatened amphibians in zoos is just 23.9% compared to over 40% in the wild. To identify key barriers to holding globally threatened amphibian species in ex situ collections, as well as potential strategies to mitigate such barriers, we surveyed amphibian curators across 107 institutions worldwide. A lack of resources (including budget, staffing and space) was perceived as the most significant barrier (87% of respondents) and the barrier most frequently identified (119 responses), followed by disease/biosecurity concerns (31 responses), and a lack of staff expertise/knowledge (30 responses). Difficulty displaying amphibians due to cryptic behaviour or colouration (65% or respondents) and difficulty attracting visitor interest (60% of respondents) were seen as insignificant barriers. Nine key priority action areas were identified, with increasing interest from zoo leadership and budget allocation identified as the most important (49% of suggested solutions). Increasing visitor interest in amphibians to encourage increased investment and engaging with range country facilities were highlighted as two ways to address barriers. Careful collection planning considering both the need and suitability of a species for captive breeding is also key, whilst critically assessing the role each species will play in a collection will enable a better assessment of the collection’s conservation value rather than using global threat status alone.     

Cd accumulation in roots and shoots of durum wheat: the roles of transpiration rate and apoplastic bypass

Cadmium is readily taken up from soils by plants, depending on soil chemistry, and variably among species and cultivars; altered transpiration and xylem transport and/or translocation in the phloem could cause this variation in Cd accumulation, some degree of which is heritable. Using Triticum turgidum var. durum cvs Kyle and Arcola (high and low grain Cd accumulating, respectively), the objectives of this study were to determine if low-concentration Cd exposure alters transpiration, to relate transpiration to accumulation of Cd in roots and shoots at several life stages, and to evaluate the role of apoplastic bypass in the accumulation of Cd in shoots. The low abundance isotope (106)Cd was used to probe Cd translocation in plants which had been exposed to elemental Cd or were Cd-na?ve; apoplastic bypass was monitored using the fluorescent dye PTS (8-hydroxy-1,3,6-pyrenetrisulphonate). Differential accumulation of Cd by 'Kyle' and 'Arcola' could be partially attributed to the effect of Cd on transpiration, as exposure to low concentrations of Cd increased mass flow and concomitant Cd accumulation in 'Kyle'. Distinct from this, exposure of 'Arcola' to low concentrations of Cd reduced translocation of Cd from roots to shoots relative to root accumulation of Cd. It is possible, but not tested here, that sequestration mechanisms (such as phytochelatin production, as demonstrated by others) are the genetically controlled difference between these two cultivars that results in differential Cd accumulation. These results also suggest that apoplastic bypass was not a major pathway of Cd transport from the root to the shoot in these plants, and that most of the shoot Cd resulted from uptake into the stele of the root via the symplastic pathway.     

Elective Delivery at Term after a Previous Unexplained Intra-Uterine Fetal Death: Audit of Delivery Outcome at Tygerberg Hospital,South Africa

Objectives

To assess the delivery outcome in a pregnancy with a previous unexplained intra-uterine death by elective induction of labour at term.

Methods

An audit of the pregnancy outcome of all women within the catchment area with a current singleton pregnancy; and a previous unexplained or unexplored singleton fetal demise ≥24 weeks (or 500 grams birth weight if gestation unknown) after planned routine induction of labour at full term (39-40 weeks).

Results

During the audit period, 306 patients with a previous intra-uterine fetal death were referred for further management. Of these, 161 had a clear indication for earlier intervention and were excluded from the protocol. Of the remaining 145 patients, 9 met further exclusion criteria and there were 2 patients who defaulted. Forty-two of the remaining study patients (with no known previous medical problems) developed complications during their antenatal course that necessitated a change in clinical management and earlier (<39 weeks) delivery. Of the remaining 92 patients in the audit, 47 (51%) went into spontaneous labour before their induction date; all 92 women delivered without major complications. There were no intra-uterine deaths prior to induction.

Conclusions

Careful follow up at a high risk clinic identifies new or concealed maternal or fetal complications in 29% of patients with a previous intra-uterine death and no obvious maternal or fetal disease in the index pregnancy. When all risks are excluded and the pregnancy allowed to progress to full term (39-40 weeks) before an induction is offered, 50% will go into spontaneous labour.