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.     

Development of an enzyme-linked immunosorbent assay for the detection and measurement of the trypanocidal drug isometamidium chloride in cattle

An enzyme-linked immunosorbent assay (ELISA) was developed to measure accurately levels of the trypanocidal drug isometamidium in the serum of treated cattle. The assay requires only 5 microliters of test serum, is sensitive to a level of 0.5 pg ml-1 and is highly specific. Cross reactivity does not occur with the two other widely used trypanocidal drugs diminazene aceturate and homidium bromide. Serum drug levels are detectable for up to six months in cattle after a single dose of 1 mg kg-1 intramuscularly, the maximum period under field conditions for which effective prophylaxis can be maintained against tsetse challenge. Application of the assay will aid the rationalisation of treatment campaigns and assist in assessing the occurrence of drug-resistant trypanosome populations.     

Ecological studies of root-nodule bacteria introduced into field environments—V.A critical examination of the stability of antigenic and streptomycin-resistance markers for identification of strains of Rhizobium tripolii

Marked strains of Rhizobium trifolii, distinguishable from other strains antigenically and by streptomycin resistance, were introduced by seed inoculation of subterranean clover (Trifolium subterraneum L.) into a field environment having a natural population of R. trifolii. Isolates from nodules obtained periodically during the following 41 months were classified using both methods of identification in parallel. This procedure made it possible to determine the reliability of each method independently.There was a gradual disappearance of the inoculum strains which occurred more rapidly in plots of cv. Woogenellup than in plots seeded with cv. Mount Barker. At five harvests, there was 95% (or greater) correspondence between inoculum survival using either method of identification. There was evidence that a small proportion of the progeny of the inocula sustained independent loss of antigenic character and/or streptomycin resistance in the field or, alternatively, that strains occurring naturally acquired these characteristics. A few nodules contained more than one strain of rhizobia. These exceptions occurred at low frequency and did not interfere substantially with identification results. It is concluded that gel immune diffusion serology and the use of streptomycin-resistant mutants are both reliable methods for identifying strains of rhizobia reisolated from field environments.     

Three-dimensional deformation measured in an alaskan glacier

Measurements of movement along 28 boreholes reveal the three-dimensional flow field in a 6 million cubic meter reach of Worthington Glacier, a temperate valley glacier located in Alaska. Sliding at the bed accounted for 60 to 70 percent of the glacier's surface motion. Strain rates in the ice were low from the surface to a depth of about 120 meters, but then increased rapidly toward the bed. Ice deformation was not affected by temporal changes in the sliding rate. The three-dimensional pattern of motion indicates that plane strain, which is often assumed by models, is a poor approximation of this viscous flow.     

Effect of post‐mortem temperature on chicken M. pectoralis major: Isometric tension and pH profiles

1. The relationship between isometric tension development and pH, as a function of storage temperature between 0° and 40°C, was examined in chicken M. pectoralis major (PM) muscle during the critical 24 h post‐mortem period.

2. The muscle strips incubated at 0°C developed a peak isometric tension of 53.3 g/cm². This occurred after only 17 min incubation when the pH was 7.02, demonstrating the potential of chicken PM muscle to cold shorten. Peak isometric tension at 5°C was considerably lower than that generated at 0°C. However, as this occurred when the muscle pH was still high (6.70), this also indicated some potential to cold shorten at 5°C.

3. At 10° to 30°C, the muscle strips developed mean peak isometric tensions of 18 g/cm² after 6 h incubation by which time the muscle pH had fallen to 6.00, demonstrating a limited potential to rigor shorten. In contrast, those incubated at 40°C developed a peak tension of 54.5 g cm² after 75 min when the muscle pH was also around 6.00, thus indicating the potential for intensive rigor shortening at this temperature. Incubation temperature and the resultant muscle pH therefore determine the potential of chicken PM muscle to either cold shorten or rigor shorten.

4. Despite the differences found in isometric tension profiles, cooked meat texture after isometric tension measurement was not significantly different at any of the temperatures studied primarily because the muscle strips were essentially prevented from shortening.     

Effect of post‐mortem temperature on chicken M. pectoralis major: Muscle shortening and cooked meat tenderness

1. Muscle shortening, sarcomere lengths and pH values were measured in strips of chicken M. pectoralis major (PM) muscle incubated at different time (0 to 24 h) and temperature (0° to 40°C) combinations immediately after slaughter; their effects on cooking loss and meat tenderness determined.

2. Maximum muscle shortening of 39% and 43% occurred at 0°C and 40°C respectively. At 0°C, most shortening occurred within 90 min postmortem when the pH of the muscle ranged from 7.13 to 6.52. In contrast, at 40°C, most shortening occurred during the development of rigor mortis, between 90 and 380 min post‐mortem, when the muscle pH ranged from 6.16 to 5.89. In a similar manner, minimum sarcomere lengths of 1.38 μm were reached after 90 min at 0°C while more severe sarcomere shortening, to 0.96 μm and 0.86 μm at 30°C and 40°C respectively, was not complete until after 380 min post‐mortem. Between 5°C and 20°C, muscle shortening ranged from 25 to 34% while minimum sarcomere lengths of 1.33 μm were recorded.

3. Cooking losses increased on average from 7 to 16% between 30 and 380 min post‐mortem, with maximum losses of 19% being achieved by the end of the 24‐h incubation period.

4. At 0°C, shear force values increased from 2.94 kg/cm² to 4.34 kg/cm² between 30 and 90 min post‐mortem while the muscle pH was > 6.5. At all other temperatures, increases in shear force values were not detected until 380 min post‐mortem when the muscle pH had fallen to 5.9 and rigor mortis had set in. At all times after 380 min, however, the muscle strips incubated at 0, 5 and 40°C had lower shear values (range 3.17 to 5.49 kg/cm²) than those incubated from 10°C to 30°C (range 5.06 to 7.22 kg/cm²).

5. A significant quadratic relationship was found between the degree of shortening and subsequent cooked meat tenderness, in which peak toughness occurred at 30% shortening. This would suggest that the actual extent of muscle shortening per se has an important role to play in determining the tenderness of chicken post‐mortem. Consequently, with unrestrained chicken muscle, where extensive shortening occurred at 0°C and 40°C (i.e cold‐ and rigor shortening) the cooked meat was more tender than that subjected to intermediate post‐mortem temperature regimens.     

Luna 9 photographs: evidence for a fragmental surface layer

The morphological features of the lunar surface photographed by Luna 9 indicate a surficial layer of weakly cohesive to noncohesive frag mental material. Most of this material is finer than a centimeter and probably finer than a few millimeters, although objects of centimeter size and larger are plentiful.     

Relationship between early post-mortem muscle pH and shortening-induced toughness in the Pectoralis major muscle of processed broilers air-chilled at 0 degrees C and -12 degrees C

1. An experiment was conducted to investigate the development of shortening-induced toughness in the Pectoralis major (PM) muscles of commercially processed broilers, air-chilled at 0 degrees C and -12 degrees C, as a function of muscle pH early post-mortem. Electrical stimulation was used immediately after stunning and neck cutting to provide carcases with pH values 15 min post-mortem (pH15 min) ranging between 6.79 and 5.85. 2. The deep PM muscle temperatures of carcases chilled at -12 degrees C were lower (cooler) after primary chilling and at 215 min post-mortem than those chilled at 0 degrees C, although chilling regimen had no major effect on pH values over the 24 h post-mortem period. However, carcases chilled at -12 degrees C had longer sarcomeres, lower cooking losses and lower shear force values than those chilled at 0 degrees C. 3. Correlation analysis of the results for both chilling regimens clearly demonstrated that over the pH15min range 6.79 to 5.85, carcases with the lowest pH15min values had the shortest sarcomeres, the highest cooking losses and the toughest meat. In addition, there was no evidence to support the occurrence of cold shortening within this population. This suggests that an early onset of rigor at higher temperatures in broiler carcases, as well as inducing rigor shortening and toughness, might also induce greater protein denaturation and subsequent loss of water holding capacity as manifested in increased cooking losses. 4. Quadratic regression curves showed that over the pH15min range 6.80 to 6.30, only the fast chilling regimen at -12 degrees C could inhibit rigor shortening and minimise changes in cooking loss and shear force values. However, neither chilling regimen was effective in preventing severe rigor shortening, increased cooking losses and adverse toughness in carcases with pH15min values below 6.30. 5. The benefits of fast chilling carcases with pH15min values above 6.3 can also be quantified in terms of carcases exceeding a 4.00 kg/cm2 toughness threshold. Only 1.9% of these carcases chilled at -12 degrees C exceeded this limit (maximum shear force value of 4.72 kg/cm2) compared to 34.9% of the carcases chilled at 0 degrees C (maximum shear force value of 8.46 kg/cm2), further emphasising the considerable reduction in textural variability and improvement in tenderness gained by fast air-chilling at -12 degrees C.