The role of survivin in angiogenesis during zebrafish embryonic development

Background  

Survivin is the smallest member of the inhibitor of apoptosis (IAP) gene family. Recently, the zebrafish survivin-1 gene has been cloned, showing remarkable sequence identity and similarity over the BIR domain compared with human and mouse survivin gene. Here we investigated the role of survivin in angiogenesis during zebrafish development. Morpholinos (MOs) targeting the 5' untranslated region (UTR) (Sur_UTR) and sequences flanking the initiation codon (Sur_ATG) of zebrafish survivin-1 gene were injected into embryos at 1–4 cell stage. Vasculature was examined by microangiography and GFP expression in Tg(fli1:EGFP) ^y1 embryos. Results: In embryos co-injected with Sur_UTR and Sur_ATG-MOs, vasculogenesis was intact but angiogenesis was markedly perturbed, especially in the inter-segmental vessels (ISV) and dorsal longitudinal anastomotic vessels (DLAV) of the trunk, the inner optic circle and optic veins of developing eyes and the sub-intestinal vessels. Apoptosis was increased, as shown by TUNEL staining and increase in caspase-3 activity. Efficacy of Sur_UTR and Sur_ATG-MOs was demonstrated by translation inhibition of co-injected 5'UTR survivin:GFP plasmids. The phenotypes could be recapitulated by splice-site MO targeting the exon2-intron junction of survivin gene and rescued by survivin mRNA. Injection of human vascular endothelial growth factor (VEGF) protein induced ectopic angiogenesis and increased survivin expression, whereas treatment with a VEGF receptor inhibitor markedly reduced angiogenesis and suppressed survivin expression. Conclusion: Survivin is involved in angiogenesis during zebrafish development and may be under VEGF regulation.     

Genetic and DNA Methylation Changes in Cotton (Gossypium) Genotypes and Tissues

In plants, epigenetic regulation is important in normal development and in modulating some agronomic traits. The potential contribution of DNA methylation mediated gene regulation to phenotypic diversity and development in cotton was investigated between cotton genotypes and various tissues. DNA methylation diversity, genetic diversity, and changes in methylation context were investigated using methylation-sensitive amplified polymorphism (MSAP) assays including a methylation insensitive enzyme (BsiSI), and the total DNA methylation level was measured by high-performance liquid chromatography (HPLC). DNA methylation diversity was greater than the genetic diversity in the selected cotton genotypes and significantly different levels of DNA methylation were identified between tissues, including fibre. The higher DNA methylation diversity (CHG methylation being more diverse than CG methylation) in cotton genotypes suggest epigenetic regulation may be important for cotton, and the change in DNA methylation between fibre and other tissues hints that some genes may be epigenetically regulated for fibre development. The novel approach using BsiSI allowed direct comparison between genetic and epigenetic diversity, and also measured CC methylation level that cannot be detected by conventional MSAP.     

Preparation, characterisation and crystal structure of two zinc(II) benzoate complexes with pyridine-based ligands nicotinamide and methyl-3-pyridylcarbamate

Two benzoate complexes namely tetrakis(μ₂-benzoato-O,O^′)-bis(μ₂-benzoato-O,O)-bis(nicotinamide-N)-tri-zinc(II), [Zn₃(benz)₆(nia)₂] (I) and bis(benzoato-O)-bis(methyl-3-pyridylcarbamate-N)-zinc(II), [Zn(benz)₂(mpcm)₂] (II) (benz=benzoate anion, nia=nicotinamide, mpcm=methyl-3-pyridylcarbamate) were prepared and characterised by elemental analysis, IR spectroscopy, thermal analysis and X-ray structure determination. The structure of the complex I is centrosymmetric, formed by a linear array of three zinc atoms. The central zinc atom shows octahedral coordination and is bridged to each of the terminal zinc atoms by three benzoate anions. Two of them act as bidentate, one as monodentate ligand. By additional coordination of the nia ligand, the terminal Zn atoms adopt tetrahedral surrounding. The structure of complex II contains two crystallographically independent [Zn(benz)₂(mpcm)₂] molecules. In each molecule, the zinc atom is tetrahedrally coordinated by two monodentate benzoate and two methyl-3-pyridylcarbamate ligands. Intermolecular hydrogen bonds of the N-H?O type connect molecules in the structures of complexes I and II to form a two-dimensional network. The three different types of carboxylate binding found in the complexes were distinguished also by values of carboxylate stretching vibrations in FT-IR spectra as well as by thermal decomposition of the complexes in nitrogen.     

Photosynthesis and Degree of Polymerization of Fructan during Reproductive Growth of Meadow Fescue at two Temperatures and two Photon Flux Densities

Accumulation of dry weight was measured in plant parts of meadowfescue (Festuca pratensis Huds.) that was grown at 16/11 °Cor 26/21 °C and with 20 or 60 nE cm^–2 s^–1 photosyntheticallyactive radiation. Plants reached anthesis about 3 weeks laterat 16/11 °C than at 26/21 °C and had then a higher proportionof dry weight in inflorescences and less in leaf blades. Growthtemperature had little effect on CO₂ exchange rate (CER) butplants grown at 60 nE cm^–2 s^–1 had higher CER thanthose grown at 20 nE cm^–2 s^–1. The concentration of water-soluble carbohydrates (WSC) at similargrowth stages was usually higher at 16/11°C than at 26/21°C.High radiation also led to higher WSC in stem and leaf tissue.Root tissue changed least and WSC did not exceed 10% of dryweight during the experiment. In all tissues, when WSC was high,the fructans were distributed into a group with a high degreeof polymerization (DP) and another with a low DP. The low DPgroup included sucrose, reducing sugars and fructans up to about20 units long. An apparent threshold concentration of WSC wasnecessary for synthesis of the high DP fructans. This concentrationwas near 12% for leaf tissue, about 6% for stem base tissue,and 2.5% for root tissue. The average apparent DP of the highDP fructan group was 43 to 50 for leaf tissue, 31 to 93 forstem base tissue, and 27 to 31 for roots. These characteristicsappeared to be mostly tissue dependent with less effect fromtemperature and radiation. Key words: Fructans, Meadow fescue, Environmental effects, Dry weight distribution     

Low genetic diversity and evidence of population structure among subspecies of Nerodia harteri, a threatened water snake endemic to Texas

Nerodia harteri is a threatened small-bodied water snake that occupies one of the most restricted ranges of any snake within the continental United States. It is found closely associated with rivers and tributaries in the Colorado and Brazos river basins, which flow through north-central Texas. Nerodia harteri has been at the center of debate owing to conflicts between conservation efforts and the construction of dams that change or destroy its preferred habitat. Additionally, its taxonomic status has also been under contention with some authors recognizing two subspecies, the Brazos water snake (N. h. harteri) and the Concho water snake (N. h. paucimaculata), whereas other authors consider each separate species. Despite its relatively recent discovery during the 1940s, N. harteri has been the subject of several ecological studies, yet no population genetic assessment of either subspecies has been performed to date. We first evaluated the phylogenetic placement of both subspecies among other North American Nerodia using partial sequence data from the mitochondrial gene cytochrome-b. We then tested for population subdivision among four rivers encompassing the range of N. harteri and tested for the presence of admixture between river basins using mitochondrial sequence data (920?bp of cyt-b) and five cross-species amplified microsatellite loci. We found low mitochondrial haplotype diversity represented by two unique haplotypes in each river basin, which were separated by no more than four nucleotide changes. Nuclear loci showed low genetic diversity and population structuring within and among river basins. We did not find conclusive evidence of admixture between basins, and we support the presence of two separate evolutionarily significant units and two separate management units corresponding to each major river basin. Given increasing natural and anthropogenic threats, we recommend continued ecological and genetic monitoring of both subspecies.     

The kinase activity of PKR represses inflammasome activity

The protein kinase R (PKR) functions in the antiviral response by controlling protein translation and inflammatory cell signaling pathways. We generated a transgenic, knock-in mouse in which the endogenous PKR is expressed with a point mutation that ablates its kinase activity. This novel animal allows us to probe the kinase-dependent and -independent functions of PKR. We used this animal together with a previously generated transgenic mouse that is ablated for PKR expression to determine the role of PKR in regulating the activity of the cryopyrin inflammasome. Our data demonstrate that, in contradiction to earlier reports, PKR represses cryopyrin inflammasome activity. We demonstrate that this control is mediated through the established function of PKR to inhibit protein translation of constituents of the inflammasome to prevent initial priming during innate immune signaling. These findings identify an important role for PKR to dampen inflammation during the innate immune response and caution against the previously proposed therapeutic strategy to inhibit PKR to treat inflammation.