Inhibition of telomerase in canine cancer cells following telomestatin treatment

Telomere shortening in normal somatic cells has been proposed as a major barrier to unlimited cellular proliferation. Telomerase is an enzyme capable of maintaining telomere length, and thus bypassing this barrier. In human beings, telomerase activity is restricted to cancer cells and cells of stem or germ cell lineages. Dogs represent a potentially useful clinical model for the development of telomerase‐based therapies because telomerase activity is also restricted to cancer cells and stem cells in this species. We examined the ability of telomestatin to inhibit telomerase activity in telomerase‐positive D17 and CMT7 canine cancer cell lines. At a concentration of 2 μM, telomestatin treatment resulted in a decrease in telomerase activity, telomere shortening, growth inhibition and apoptosis in telomerase‐positive cancer cells. These effects were not seen in telomerase‐negative skin fibroblasts or negative controls. These results confirm that telomestatin specifically inhibits telomerase activity in canine cancer cells and strengthens the usefulness of dogs as a model for testing telomerase‐based therapies.     

Preparation and evaluation of antigens used in serological tests for caprine syncytial retrovirus antibody in sheep and goat sera

Methods used to prepare antigens from caprine syncytial retrovirus (CSR) for use in the agarose gel immunodiffusion test (AGID) or an indirect enzyme-linked immunosorbent assay (ELISA) are described. Caprine and ovine sera were tested for antibody to CSR using the AGID test and ELISA incorporating a caprine system (CSR antigen and rabbit anti-goat IgG) or an ovine system (maedi-visna virus antigen and rabbit anti-sheep IgG). Good correlation was achieved in the results of the 3 tests when sera were devoid of antibody or were strongly positive. Variations in the results on weakly positive sera were considered to be more a matter of interpretation than due to basic differences in the reagents employed.     

Dose-titration studies of ivermectin against experimental Ancylostoma caninum and Uncinaria stenocephala infections

Dose-titration trials of ivermectin were conducted on pups with dual experimental infections of 4th-stage larvae or adult Ancylostoma caninum and Uncinaria stenocephala. Ivermectin was administered orally or subcutaneously at dosages of 0.006, 0.012, or 0.024 mg/kg of body weight. Maximal or near maximal (greater than or equal to 96% to 100%) anthelmintic effect was observed for both stages of development for each hookworm species by either route of administration at a dosage of 0.024 mg/kg. Responses for all of the aforementioned categories were linearly related to increasing log dosage of ivermectin, with common slopes (regression coefficients). Regression analysis also provided estimates of the minimal dosages required to produce maximal reduction in worm burden for each stage, species, and route of administration. The estimated ivermectin dosages for maximal efficacy ranged from a low of 0.014 mg/kg for adult A caninum by oral treatment to 0.044 mg/kg for 4th-stage larvae of U stenocephala by oral treatment.     

Genetic improvement of the Pacific oyster Crassostrea gigas (Thunberg) in Australia

The Pacific oyster industry in Australia is derived from importations from Japan in the late 1940s and early 1950s to Tasmania and is almost completely hatchery based. This makes it a good target for developing and deploying genetically improved strains. An allozyme survey comparing hatchery stocks with self‐recruiting Tasmanian stocks and with two collections from Japan found abundant variation and no significant evidence of allele loss. The subsequent selection programme (initiated in the summer of 1996/97) had several strands. We wanted to take advantage of the increased power that marker‐assisted selection could bring and, therefore, needed to develop a linkage map and isolate flanking markers around quantitative trait loci (QTLs). Several types of markers (allozymes, microsatellites and AFLPs) were used, and single‐pair crosses were set up; QTLs have been detected. Conventional selection programmes, one based on mass selection and one on family selection, have been established. Triploid Pacific oysters produced via chemical means have been available for several years, but rates of triploidy achieved by such means are usually less than 100%. In 1999, we will assess whether our tetraploid 2‐year‐old broodstock can be crossed with diploids to give 100% triploid offspring.     

Transpiration-induced axial and radial tension gradients in trunks of Douglas-fir trees

We determined the axial and radial xylem tension gradients in trunks of young Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) trees. Axial specific conductivity (k(s-a)) and sap flux density (Js) were measured at four consecutive depths within the sapwood at a stem height of 1 m. By definition, at a given position in the bole, Js is a function not only of k(s-a) but also of the driving force for water movement. The Js:k(s-a) ratio was therefore used to estimate axial tension gradients and the radial gradients at a stem height of 1 m were calculated from the differences in axial tension gradients at each depth. Tracheid lumen diameter and tracheid length were used to predict differences in k(s-a) and its divergence from the theoretical k(s-a) determined by the Hagen Poisseuille equation. The ratio of k(s-a) (determined in the laboratory) to Js (measured in the field) varied with depth in the sapwood, resulting in non-uniform axial and radial tension gradients from inner to outer sapwood. Transpiration-induced axial tension gradients were in the range of 0.006-0.01 MPa m(-1) excluding the gravitational tension gradient. At a stem height of 1 m, radial tension gradients were in the range of 0.15-0.25 MPa m(-1) and were lower in the middle sapwood than in the inner or outer sapwood. Axial tension gradients were 44-50% higher in the outer sapwood than in the inner sapwood. At a stem height of 1 m, radial Js, calculated on the basis of radial tension gradients and measured radial specific conductivity (k(s-r)), was about two orders of magnitude smaller than axial Js. Our findings indicate that large radial tension gradients occur in the sapwood and clarify the role played by xylem k(s-a) and k(s-r) in determining in situ partitioning of Js in the axial and radial directions.     

Effect of Steeping Conditions on Wet-Milling Characteristics of Hard Red Winter Wheat

A batch-wise small-scale wet-processing laboratory for whole wheat kernel has been designed and constructed to produce wheat starch and gluten from wheat grains. Hard red winter wheat kernels were steeped in three steeping media: SO₂ solution, lactic acid, and hydrochloric acid. Acid concentrations of 0.1, 0.3, and 0.5%, were used for SO₂ solutions and hydrochloric acid, and 0.1, 0.6, and 3.0% for lactic acid. After 16, 20, and 24 hr of steeping, the wheat was wet-milled. Yields and protein contents of wet-milling fractions were compared. Both high concentration of steeping media and long steeping time increased the starch yield and decreased the protein contents of the starch. However, the steeping time and acid concentration could be reduced from 24 to 20 hr and from 0.5 to 0.3%, respectively, without any statistically significant difference in starch yields or protein contents of the starch. Consistency and color of the starch were affected by both steeping time and acid concentrations of steeping media.