Structural basis of type II topoisomerase inhibition by the anticancer drug etoposide

Type II topoisomerases (TOP2s) resolve the topological problems of DNA by transiently cleaving both strands of a DNA duplex to form a cleavage complex through which another DNA segment can be transported. Several widely prescribed anticancer drugs increase the population of TOP2 cleavage complex, which leads to TOP2-mediated chromosome DNA breakage and death of cancer cells. We present the crystal structure of a large fragment of human TOP2β complexed to DNA and to the anticancer drug etoposide to reveal structural details of drug-induced stabilization of a cleavage complex. The interplay between the protein, the DNA, and the drug explains the structure-activity relations of etoposide derivatives and the molecular basis of drug-resistant mutations. The analysis of protein-drug interactions provides information applicable for developing an isoform-specific TOP2-targeting strategy.     

Foliar nematode,Litylenchus crenatae ssp. mccannii,population dynamics in leaves and buds of beech leaf disease‐affected trees in Canada and the US

A foliar nematode, Litylenchus crenatae ssp. mccannii, is associated with beech leaf disease (BLD) symptoms. Information about the types of tissues parasitized and how nematode populations fluctuate in these tissues over time is needed to improve surveys as well as understand the nematodes role in BLD. During this study, the nematode was detected throughout the known range of BLD by researchers at both Canadian and US institutions using a modified pan method to extract nematodes. Monthly collections of symptomatic and asymptomatic leaves during the growing season (May–October), and leaves and buds between growing seasons (November–March), revealed that nematodes were present in all tissue types. Progressively larger numbers of nematodes were detected in symptomatic leaves from Ohio and Ontario, with the greatest detections at the end of the growing season. Smaller numbers of nematodes were detected in asymptomatic leaves from BLD‐infected trees, typically at the end of the growing season. The nematode was detected overwintering in buds and detached leaves. The discovery of small numbers of nematodes in detached leaves at one location before BLD was detected indicates that nematodes may have been present before disease symptoms were expressed. Other nematodes, Plectus and Aphelenchoides spp., were infrequently detected in small numbers. Our findings support the involvement of the nematode in BLD and indicate that symptoms develop only when certain requirements, such as infection of buds, are met. We also found that the nematode can be reliably detected in buds and leaves using the modified pan extraction method.     

Apium graveolens PI 181714 is a source of resistance to Fusarium oxysporum f. sp. apii race 4 in celery (A. graveolens var. dulce)

Celery has little genetic diversity and is highly susceptible to the new fungal pathogen Fusarium oxysporum f. sp. apii (Foa) race 4. After screening an Apium graveolens germplasm collection for resistance to Foa race 4, we crossed celery cv. 'Challenger', which is Foa race 2-resistant but Foa race 4-susceptible and A. graveolens PI 181714, which is Foa races 2- and 4-resistant but non-celery type. After selfing F₁s, we screened the F₁S₁ for race 4-resistance and celery-type and then selfed selected F₁S₁. Greenhouse and field trials indicate that three selected F₁S₂ families (76–8-4, 76–8-27 and 76–8-36) are suitable as germplasm for celery breeders for resistance to Foa race 4. A F1S3 76–8–36-124 is either fixed or nearly so for resistance to Foa races 4 and 2. Furthermore, quantitative PCR indicates that PI 181714 is resistant, rather than tolerant, to Foa races 4 and 2, and that this resistance has been introgressed into F1S3 76–8–36-124.     

Using EM and VERIS technology to assess land suitability for orchard and vineyard development

Orchard and vineyard producers conduct preplant site evaluations to help prevent planting permanent tree and vine crops on lands where the crop will not perform to its highest potential or attain its full life expectancy. Physical soil characteristics within specific soil profiles and spatially throughout an orchard influence decisions on land preparation, irrigation system selection, horticultural choices, and nutrient management. Producers depend on soil surveys to help them understand the soil characteristics of the land and may be interested in technology that provides additional information. Electromagnetic induction (EM38) and four-probe soil resistance sensors (VERIS) are being used in combination with global positioning systems to map spatial variability of soils using apparent soil electrical conductivity (ECa). The hypothesis evaluated in this study is whether rapid, in situ, and relatively low-cost methods of measuring ECa (EM38 and VERIS) can effectively identify and map physical soil variability in non-saline soils. The supposition is that in non-saline soils, ECa levels will relate well to soil texture and water-holding capacity and can be used to map physical soil variability. In turn, the information can be used to guide decisions on preplant tillage, irrigation system design, water and nutritional management, and other horticultural considerations. Two sites in the Sacramento Valley were mapped each with EM38 and VERIS methods. Site-specific management zones were identified by each provider on ECa maps for each site, and then soil samples were collected by University of California researchers to verify these zones. Results showed that on non-saline soils, ECa measured with both EM38 and VERIS correlate with physical soil properties such as gravel, sand, silt, and clay content but the relationship between conductivity and these physical soil properties varied from moderately strong to weak. The strength of the correlation may be affected by several factors including how dominant soil texture is on conductivity relative to other soil properties and on methods of equipment operation, data analysis and interpretation. Overall, the commercial providers of ECa surveys in this study delivered reasonable levels of accuracy that were consistent with results reported in previous studies. At one site, an ECa map developed with VERIS provided more detail on physical soil variability to supplement published soil surveys and aided in the planning and development of a walnut orchard. At a second site, almond yield appeared to correlate well with distinctly different soil zones identified with EM38 mapping.     

Preparation of tetrahydroagathic acid: a serum metabolite of isocupressic acid, a cattle abortifacient in ponderosa pine

Isocupressic acid (1) was used to synthetically prepare a mixture of (8S,13R,S)-labda-15,19-dioic acid (tetrahydroagathic acid) (5) via a two-step oxidation procedure followed by hydrogenation of the double bonds at C13 and C8. Reduction of the C8,17 double bond was stereospecific producing only the 8S isomer and confirmed by the nOe interaction between the resulting C17 and C20 methyl groups. The 13R and 13S isomers of 5 were separated and analyzed by HPLC/MS, and (13S)-tetrahydroagathic acid was isolated and identified by comparison to a standard prepared by hydrogenation of naturally occurring (13S)-dihydroagathic acid (4). (13R,S)-dihydroagathic acid was prepared by selective sodium metal-catalyzed hydrogenation of the C13,14 allylic double bond of agathic acid (3). The prepared compounds were then used as standards to confirm the presence of 4 and 5 and their respective 13R and 13S isomers in bovine serum samples. Tetrahydroagathic acid was shown to be the only metabolite detected in serum samples taken from a suspected cattle abortion case submitted for diagnosis; and, thus, 5 could be a valuable diagnostic marker for pine needle-induced abortions.