Using crossover breakpoints in recombinant inbred lines to identify quantitative trait loci controlling the global recombination frequency

Recombination is a crucial component of evolution and breeding, producing new genetic combinations on which selection can act. Rates of recombination vary tremendously, not only between species but also within species and for specific chromosomal segments. In this study, by examining recombination events captured in recombinant inbred mapping populations previously created for maize, wheat, Arabidopsis, and mouse, we demonstrate that substantial variation exists for genomewide crossover rates in both outcrossed and inbred plant and animal species. We also identify quantitative trait loci (QTL) that control this variation. The method that we developed and employed here holds promise for elucidating factors that regulate meiotic recombination and for creation of hyperrecombinogenic lines, which can help overcome limited recombination that hampers breeding progress.     

Lead optimization of methionine aminopeptidase-2 (MetAP2) inhibitors containing sulfonamides of 5,6-disubstituted anthranilic acids

A series of aryl sulfonamides of 5,6-disubstituted anthranilic acids were identified as potent inhibitors of methionine aminopeptidase-2 (MetAP2). Small alkyl groups and 3-furyl were tolerated at the 5-position of anthranilic acid, while -OCH(3), CH(3), and Cl were found optimal for the 6-position. Placement of 2-aminoethoxy group at the 6-position enabled interaction with the second Mn(2+) but did not result in enhancement in potency. Introduction of a tertiary amino moiety at the ortho-position of the sulfonyl phenyl ring gave reduced protein binding and improved cellular activity, but led to lower oral bioavailability.     

Thienopyridine urea inhibitors of KDR kinase

A series of substituted thienopyridine ureas was prepared and evaluated for enzymatic and cellular inhibition of KDR kinase activity. Several of these analogs, such as 2, are potent inhibitors of KDR (<10 nM) in both enzymatic and cellular assays. Further characterization of inhibitor 2 indicated that this analog possessed excellent in vivo potency (ED50 2.1 mg/kg) as measured in an estradiol-induced mouse uterine edema model.     

Discovery of hidden pedigree errors combining genomic information with the genomic relationship matrix in Texel sheep

Genomic variants such as Single Nucleotide Polymorphisms and animal pedigree are now used widely in routine genetic evaluations of livestock in many countries. The use of genomic information not only can be used to enhance the accuracy of prediction but also to verify pedigrees for animals that are extensively managed using natural mating and enabling multiple-sire mating groups to be used. By so doing, the rate of genetic gain is enhanced, and any bias associated with incorrect pedigrees is removed. This study used a set of 8 764 sheep genotypes to verify the pedigree based on both the conventional opposing homozygote method as well as a novel method when combined with the inclusion of the genomic relationship matrix (GRM). The genomic relationship coefficients between verified pairs of animals showed on average a relationship of 0.50 with parent, 0.25 with grandparent, 0.13 with great grandparent, 0.50 with full-sibling and 0.27 with half-sibling. Minimum obtained values from these verified pairs were then used as thresholds to determine the pedigree for unverified pairs of animals, to detect potential errors in the pedigree. Using a case study from a population partially genotyped UK sheep, the results from this study illustrate a powerful way to resolve parentage inconsistencies, when combining the conventional ‘opposing homozygote’ method using genomic information together with GRM for pedigree checking. In this way, previously undetected pedigree errors can be resolved.     

Patterns and drivers of stream benthic macroinvertebrate beta diversity in an agricultural landscape

Hydrobiologia - Laguna del Sauce, one of the main drinking water sources in Uruguay, is an eutrophic shallow lake with high temporal variation of inorganic turbidity caused by extreme wind events....     

Zebrafish adult pigment stem cells are multipotent and form pigment cells by a progressive fate restriction process

Skin pigment pattern formation is a paradigmatic example of pattern formation. In zebrafish, the adult body stripes are generated by coordinated rearrangement of three distinct pigment cell‐types, black melanocytes, shiny iridophores and yellow xanthophores. A stem cell origin of melanocytes and iridophores has been proposed although the potency of those stem cells has remained unclear. Xanthophores, however, seemed to originate predominantly from proliferation of embryonic xanthophores. Now, data from Singh et al. shows that all three cell‐types derive from shared stem cells, and that these cells generate peripheral neural cell‐types too. Furthermore, clonal compositions are best explained by a progressive fate restriction model generating the individual cell‐types. The numbers of adult pigment stem cells associated with the dorsal root ganglia remain low, but progenitor numbers increase significantly during larval development up to metamorphosis, likely via production of partially restricted progenitors on the spinal nerves.     

Grazing effects on plant cover,soil and microclimate in fragmented woodlands in south‐western Australia: implications for restoration

This study investigated the impacts of livestock grazing on native plant species cover, litter cover, soil surface condition, surface soil physical and chemical properties, surface soil hydrology, and near ground and soil microclimate in remnant Eucalyptus salmonophloia F. Muell woodlands. Vegetation and soil surveys were undertaken in three woodlands with a history of regular grazing and in three woodlands with a history of little or no grazing. Livestock grazing was associated with a decline in native perennial cover and an increase in exotic annual cover, reduced litter cover, reduced soil cryptogam cover, loss of surface soil microtopography, increased erosion, changes in the concentrations of soil nutrients, degradation of surface soil structure, reduced soil water infiltration rates and changes in near ground and soil microclimate. The results suggest that livestock grazing changes woodland conditions and disrupts the resource regulatory processes that maintain the natural biological array in E. salmonophloia woodlands. Consequently the conditions and resources in many remnant woodlands may be above or below critical thresholds for many species. The implications of these findings for restoration of plant species diversity and community structure are discussed. Simply removing livestock from degraded woodlands is unlikely to result in the restoration of plant species diversity and community structure. Restoration will require strategies that capture resources, increase their retention and improve microclimate.     

Hexokinase 3 enhances myeloid cell survival via non-glycolytic functions

The family of hexokinases (HKs) catalyzes the first step of glycolysis, the ATP-dependent phosphorylation of glucose to glucose-6-phosphate. While HK1 and HK2 are ubiquitously expressed, the less well-studied HK3 is primarily expressed in hematopoietic cells and tissues and is highly upregulated during terminal differentiation of some acute myeloid leukemia (AML) cell line models. Here we show that expression of HK3 is predominantly originating from myeloid cells and that the upregulation of this glycolytic enzyme is not restricted to differentiation of leukemic cells but also occurs during ex vivo myeloid differentiation of healthy CD34⁺ hematopoietic stem and progenitor cells. Within the hematopoietic system, we show that HK3 is predominantly expressed in cells of myeloid origin. CRISPR/Cas9 mediated gene disruption revealed that loss of HK3 has no effect on glycolytic activity in AML cell lines while knocking out HK2 significantly reduced basal glycolysis and glycolytic capacity. Instead, loss of HK3 but not HK2 led to increased sensitivity to ATRA-induced cell death in AML cell lines. We found that HK3 knockout (HK3-null) AML cells showed an accumulation of reactive oxygen species (ROS) as well as DNA damage during ATRA-induced differentiation. RNA sequencing analysis confirmed pathway enrichment for programmed cell death, oxidative stress, and DNA damage response in HK3-null AML cells. These signatures were confirmed in ATAC sequencing, showing that loss of HK3 leads to changes in chromatin configuration and increases the accessibility of genes involved in apoptosis and stress response. Through isoform-specific pulldowns, we furthermore identified a direct interaction between HK3 and the proapoptotic BCL-2 family member BIM, which has previously been shown to shorten myeloid life span. Our findings provide evidence that HK3 is dispensable for glycolytic activity in AML cells while promoting cell survival, possibly through direct interaction with the BH3-only protein BIM during ATRA-induced neutrophil differentiation.Subject terms: Cell biology, Cancer     

A hybrid method for peptide identification using integer linear optimization, local database search, and quadrupole time-of-flight or OrbiTrap tandem mass spectrometry

A novel hybrid methodology for the automated identification of peptides via de novo integer linear optimization, local database search, and tandem mass spectrometry is presented in this article. A modified version of the de novo identification algorithm PILOT, is utilized to construct accurate de novo peptide sequences. A modified version of the local database search tool FASTA is used to query these de novo predictions against the nonredundant protein database to resolve any low-confidence amino acids in the candidate sequences. The computational burden associated with performing several alignments is alleviated with the use of distributive computing. Extensive computational studies are presented for this new hybrid methodology, as well as comparisons with MASCOT for a set of 38 quadrupole time-of-flight (QTOF) and 380 OrbiTrap tandem mass spectra. The results for our proposed hybrid method for the OrbiTrap spectra are also compared with a modified version of PepNovo, which was trained for use on high-precision tandem mass spectra, and the tag-based method InsPecT. The de novo sequences of PILOT and PepNovo are also searched against the nonredundant protein database using CIDentify to compare with the alignments achieved by our modifications of FASTA. The comparative studies demonstrate the excellent peptide identification accuracy gained from combining the strengths of our de novo method, which is based on integer linear optimization, and database driven search methods.