Predisposition to cancer caused by genetic and functional defects of mammalian Atad5

ATAD5, the human ortholog of yeast Elg1, plays a role in PCNA deubiquitination. Since PCNA modification is important to regulate DNA damage bypass, ATAD5 may be important for suppression of genomic instability in mammals in vivo. To test this hypothesis, we generated heterozygous (Atad5(+/m)) mice that were haploinsuffficient for Atad5. Atad5(+/m) mice displayed high levels of genomic instability in vivo, and Atad5(+/m) mouse embryonic fibroblasts (MEFs) exhibited molecular defects in PCNA deubiquitination in response to DNA damage, as well as DNA damage hypersensitivity and high levels of genomic instability, apoptosis, and aneuploidy. Importantly, 90% of haploinsufficient Atad5(+/m) mice developed tumors, including sarcomas, carcinomas, and adenocarcinomas, between 11 and 20 months of age. High levels of genomic alterations were evident in tumors that arose in the Atad5(+/m) mice. Consistent with a role for Atad5 in suppressing tumorigenesis, we also identified somatic mutations of ATAD5 in 4.6% of sporadic human endometrial tumors, including two nonsense mutations that resulted in loss of proper ATAD5 function. Taken together, our findings indicate that loss-of-function mutations in mammalian Atad5 are sufficient to cause genomic instability and tumorigenesis.     

Differential expressions of adhesive molecules and proteases define mechanisms of ovarian tumor cell matrix penetration/invasion

Epithelial ovarian cancer is an aggressive and deadly disease and understanding its invasion mechanisms is critical for its treatment. We sought to study the penetration/invasion of ovarian tumor cells into extracellular matrices (ECMs) using a fibroblast-derived three-dimensional (3D) culture model and time-lapse and confocal imaging. Twelve ovarian tumor cells were evaluated and classified into distinct groups based on their ECM remodeling phenotypes; those that degraded the ECM (represented by OVCAR5 cells) and those that did not (represented by OVCAR10 cells). Cells exhibiting a distinct ECM modifying behavior were also segregated by epithelial- or mesenchymal-like phenotypes and uPA or MMP-2/MMP-9 expression. The cells, which presented epithelial-like phenotypes, penetrated the ECM using proteases and maintained intact cell-cell interactions, while cells exhibiting mesenchymal phenotypes modified the matrices via Rho-associated serine/threonine kinase (ROCK) in the absence of apparent cell-cell interactions. Overall, this study demonstrates that different mechanisms of modifying matrices by ovarian tumor cells may reflect heterogeneity among tumors and emphasize the need to systematically assess these mechanisms to better design effective therapies.     

Synthesis and fungicidal activity of tubulin polymerisation promoters. Part 2: pyridazines

Special tetrasubstituted pyridazines are potent fungicides by promoting the tubulin polymerisation, hereby disrupting the microtubule dynamics in the fungus. They are monocyclic analogs of similar substituted triazolopyrimidines and pyridopyrazines with the same mode of action. The fungicidal activity of these pyridazines was evaluated against the plant pathogens Botrytis cinerea (grey mould), Mycosphaerella graminicola (wheat leaf blotch) and Alternaria solani (potato and tomato early blight). Structure-activity relationship studies revealed the importance of a methyl and a chlorine substituent next to both ring nitrogen atoms and two aryl or heteroaryl groups in the other two pyridazine positions.     

Highly efficient sorghum transformation

A highly efficient microprojectile transformation system for sorghum (Sorghum bicolor L.) has been developed by using immature embryos (IEs) of inbred line Tx430. Co-bombardment was performed with the neomycin phosphotransferase II (nptII) gene and the green fluorescent protein (gfp) gene, both under the control of the maize ubiquitin1 (ubi1) promoter. After optimization of both tissue culture media and parameters of microprojectile transformation, 25 independent transgenic events were obtained from 121 bombarded IEs. The average transformation frequency (the total number of independent transgenic events divided by the total number of bombarded IEs) was 20.7% in three independent experiments. Transgenic events were confirmed by both PCR screening and Southern hybridization of genomic DNA from primary transgenics (T?). More than 90% of transformants were fertile and displayed normal morphology in a containment glasshouse. Co-transformation rate of the nptII and gfp genes was 72% in these experiments. The segregation of nptII and gfp in T? progenies was observed utilizing fluorescence microscopy and geneticin selection of seedlings indicating both were inherited in the T? generation. The transformation procedure, from initiating IEs to planting putative transgenic plantlets in the glasshouse, was completed within 11-16 weeks, and was approximately threefold more efficient than the previously reported best sorghum transformation system.     

The Ngorongoro Volcanic Highland and its relationships to volcanic deposits at Olduvai Gorge and East African Rift volcanism

The Ngorongoro Volcanic Highland (NVH), situated adjacent and to the east of Olduvai Gorge in northern Tanzania, is the source of the immense quantities of lava, ignimbrite, air fall ash, and volcaniclastic debris that occur interbedded in the Plio-Pleistocene sedimentary deposits in the Laetoli and Olduvai areas. These volcanics have proven crucial to unraveling stratigraphic correlations, the age of these successions, the archaeological and paleontological remains, as well as the source materials from which the bulk of the stone tools were manufactured. The NVH towers some 2,000 m above the Olduvai and Laetoli landscapes, affecting local climate, run-off, and providing varying elevation - climate controlled ecosystem, habitats, and riparian corridors extending into the Olduvai and Laetoli lowlands. The NVH also plays a crucial role in addressing the genesis and history of East African Rift (EAR) magmatism in northern Tanzania.In this contribution, we provide age and petrochemical compositions of the major NVH centers: Lemagurut, basalt to benmorite, 2.4-2.2 Ma; Satiman, tephrite to phonolite, 4.6-3.5 Ma; Oldeani, basalt to trachyandesite, 1.6-1.5 Ma; Ngorongoro, basalt to rhyolite, 2.3-2.0 Ma; Olmoti, basalt to trachyte, 2.0-1.8 Ma; Embagai, nephelinite to phonolite, 1.2-0.6 Ma; and Engelosin, phonolite, 3-2.7 Ma. We then discuss how these correlate in time and composition with volcanics preserved at Olduvai Gorge. Finally, we place this into context with our current understanding as to the eruptive history of the NVH and relationship to East African Rift volcanism.     

Concentrations and Distribution of Trace Metals in Water and Streambed Sediments of Orogodo River,Southern Nigeria

The distribution of Cd, Pb, Ni, Cr, Cu, Mn, Fe, and Zn in sediment and surface water, and some physico-chemical characteristics of Orogodo river sediments, were evaluated. The sediment pH ranged from 5.1–7.3; conductivity values ranged from 34.5 to 389.0 μScm^?1. Total nitrogen values ranged from 0.06–0.10%, NH₃-N values ranged from 0.25–0.44 mgkg^?1, percent total organic carbon ranged from 0.21–1.68%, and total phosphorus values ranged from 0.004–0.02% for dry and wet seasons. The sand fraction consists of 87–95%, silt fractions ranged from 0–2%, and clay fraction between 4–13%. The mean concentrations of metals (dry weight basis) in the streambed sediments ranged from 1.92–17.37 mgkg^?1 for Cu, 0.98–4.78 mgkg^?1 for Ni, 0.01–32.98 mgkg^?1 for Mn, 353.22–2045.64 mgkg^?1 for Fe, 69.96–100.16 mgkg^?1 for Zn, 0.21–1.32 mgkg^?1 for Cr, and Cd was less than 0.001 mgkg^?1 for wet and dry seasons. The mean concentrations of metals in the surface water ranged between 0.01–0.05–0.05 mg/L for Cu, nd-0.11 mg/L for Ni, 0.001–0.31 mg/L for Pb, 0.001–1.82 mg/L Mn, 0.01–3.52 mg/L for Fe, 0.16–0.61 mg/L for Zn, nd-0.007 mg/L for Cr, and <0.001 mg/L for Cd. Based on principal component analysis, two main sources of metals in the Orogodo River can be identified: (i) Cr, Cu, Pb, and Fe are mainly derived from industrial sources; (ii) Mn, Zn, and Ni associated with traffic activities. No element examined had a contamination/pollution index value greater than unity (pollution ranges). This implies that the multiple pollution indices obtained from the analysis showed that Orogodo River sediments were not polluted with heavy metals.     

Allelic variation of the β-, γ- and δ-kafirin genes in diverse Sorghum genotypes

The β-, γ- and δ-kafirin genes were sequenced from 35 Sorghum genotypes to investigate the allelic diversity of seed storage proteins. A range of grain sorghums, including inbred parents from internationally diverse breeding programs and landraces, and three wild Sorghum relatives were selected to encompass an extensive array of improved and unimproved germplasm in the Eusorghum. A single locus exists for each of the expressed kafirin-encoding genes, unlike the multigenic α-kafirins. Significant diversity was found for each locus, with the cysteine-rich β-kafirin having four alleles, including the first natural null mutant reported for this prolamin subfamily. This allele contains a frame shift insertion at +206 resulting in a premature stop codon. SDS-PAGE revealed that lines with this allele do not produce β-kafirin. An analysis of flour viscosity reveals that these β-kafirin null lines have a difference in grain quality, with significantly lower viscosity observed over the entire Rapid ViscoAnalyser time course. There was less diversity at the protein level within the cysteine-rich γ-kafirin, with only two alleles in the cultivated sorghums. There were only two alleles for the δ-kafirin locus among the S. bicolor germplasm, with one allele encoding ten extra amino acids, of which five were methionine residues, with an additional methionine resulting from a nucleotide substitution. This longer allele encodes a protein with 19.1% methionine. The Asian species, S. propinquum, had distinct alleles for all three kafirin genes. We found no evidence for selection on the three kafirin genes during sorghum domestication even though the δ-kafirin locus displayed comparatively low genetic variation. This study has identified genetic diversity in all single copy seed storage protein genes, including a null mutant for β-kafirin in Sorghum.