Identification of Cytokeratins in Bovine Sperm Outer Dense Fibre Fractions

Outer dense fibres (ODF) are important substructures of mammalian sperm tails that are involved in the regulation of sperm motility. In this study, we investigated the identity of several sodium dodecyl sulphate (SDS)‐insoluble ODF proteins. Bovine ODF were purified by separating sperm heads and tails using ultrasound and Percoll^® density gradient centrifugation. Sperm flagella were treated with the detergent cetyltrimethylammonium bromide (CTAB). CTAB‐insoluble material, which reportedly represents the ODF fraction, was collected, and electron microscopy confirmed a highly purified ODF fraction. We found after solubilization of this fraction with SDS that high amounts of insoluble material were retained after centrifugation. SDS‐insoluble material was collected and quantitatively dissolved in 8 M urea. SDS‐gel electrophoresis in the presence of urea revealed polypeptides with apparent molecular masses of approximately 25, 43, and 50 kDa. Subsequent immunoblotting with anti‐cytokeratin antibodies detected two urea‐soluble, SDS‐insoluble proteins with apparent molecular masses of 45 and 66 kDa. The 45‐kDa protein was identified as cytokeratin 19. An antibody reacting with a palette of cytokeratins (CK 1–18 and CK 20), KL1, was the only antibody that reacted with the 66‐kDa polypeptide. We conclude that sperm ODF fractions contain at least one each of type I and type II intermediate filaments. As keratins and intermediate filaments are described as rope‐like structures, we suggest that these intermediate filaments play an important structural or tension‐bearing role in sperm flagella.     

Effects of Confluent, Roscovitine Treatment and Serum Starvation on the Cell-cycle Synchronization of Bovine Foetal Fibroblasts

The present study was designed to examine the effects of cell-cycle synchronization protocols, such as confluent, roscovitine treatment and serum starvation, in bovine foetal fibroblasts on synchronization accuracy at G0/G1, viability, apoptosis, necrosis and ploidy for use as a nuclei donor. The cells in 5-10 passages were randomly allocated into three treated groups. Cells were cultured either in Dulbecco's modified Eagle's medium (DMEM) + 10% foetal bovine serum (FBS) until 90% confluent (group 1, confluent), in DMEM + 10% FBS + 30 microM roscovitine for 12 h (group 2, roscovitine), or in DMEM + 0.5% FBS for 5 days (group 3, serum starvation). Most of the cells (>80%) in all groups were arrested at the G0/G1 stage. Although the rates did not differ, cells in group 1 showed an increased cell population arrested at the G0/G1 phase. Significantly (p < 0.05) higher rates of apoptosis occurred in group 3 than in group 1 and 2 (10% vs 6% and 6%, respectively). No differences in chromosomal abnormality were observed among groups. However, by increasing the number of cell culture passages up to 15, significantly (p < 0.05) higher chromosomal abnormality was observed than in 5 and 10 passages (39% vs 28% and 23%, respectively) in group 1. The results clearly indicated that bovine foetal fibroblasts could be effectively synchronized at G0/G1 stages by all the three different treatments, confluent, roscovitine and serum starvation. However, cells in confluent showed reduced apoptosis and necrosis when they underwent 5-10 passages, exhibiting increased percentage of cells with stable chromosome diversity. Hence, cells in confluent merit further studies before they could be used as nuclear donors.     

Studies on the chemical control of blackgrass (Alopecurus myosuroides Huds.) in winter wheat in the West Midlands

In trials carried out in 1978–80, blackgrass populations in winter wheat growing on heavy soil were substantially reduced by all chemical treatments tested, the least effective being methabenzthiazuron. Wild oat populations were also reduced by all treatments except chlofop-isobutyl and methabenzthiazuron. On average of the 3 years only metoxuron treatment gave wheat yields significantly higher than the unsprayed crop.     

Theileria orientalis: a review

Theileria orientalis (also known historically as T. sergenti and T. buffeli) is responsible for benign or non-transforming theileriosis, and exerts its major effect through erythrocyte destruction. The life cycle of T. orientalis is essentially similar to that of other Theileria species, except that the schizonts do not induce transformation and fatal lymphoproliferation. The pathogenesis of anaemia as a result of infection is not clearly established and may be multifaceted. Clinical signs of weakness, reluctance to walk and abortion are early but non-specific indications of disease, particularly if accompanied by a history of cattle being moved. Physical examination may reveal pallor (pale eyes, vaginal mucosa), pyrexia, and elevated heart and respiratory rates. T. orientalis is an economically important parasite of cattle in New Zealand, Australia and Japan, especially where naïve animals are introduced into an endemic area or in animals under stress. Increased awareness of the risks posed by the parasite is required to enable management practices to be implemented to minimise its impact.     

The solid solution equilibria of lead and cadmium in polluted soils

A method for the measurement of Pb and Cd in equilibrium soil solutions involving soil equilibration with a dilute Ca electrolyte, centrifugation and filtration to <0.2 μm was evaluated. The procedure was subsequently used for the analysis of 100 Pb- and 30 Cd-contaminated soils. Solutions were analysed for Pb- and Cd using graphite-furnace AAS and the concentrations of Pb²⁺ and Cd²⁺ were estimated using standard speciation calculations. The concentrations of Pb and Cd found in the soil solutions were in the range 3.5–3600 μg dmp ^?3 and 2.7–1278 μg dm ^?3 respectively; both ranges represented less than 0.1% of the total metal concentration in the soils. Depending on solution pH, Pb ⁺² accounted for between 42–78% of Pb in solution while about 65% of Cd in solution was present as Cd⁺². The concentrations of Pb²⁺ and Cd²⁺ in solution suggested that the soil solutions were undersaturated with respect to the solid phases PbC0₃ and CdC0₃ but supersaturated with respect to Pb₅(P0₄)₃Cl and, for some samples, Cd₃(P0₄)₂ respectively. However, for both metals, a good empirical relationship was obtained between the total metal concentration in soil (mol kg^?1), free metal concentration in solution (mol dm^?3) and solution pH. The relationships took the general form of a pH-dependent Freundlich adsorption equation: For both lead and cadmium relationships, the values ofn and K₁ were close to unity, so that the distribution coefficient could be estimated from pH and a single metal-dependent constant, K₂. The algorithms appeared to be valid over a metal concentration range of four logarithmic units and pH range of 3.5–7.5.     

含莫纳可林K红曲降低鸡蛋胆固醇效果的研究

48只68周龄的海兰褐蛋鸡分成三组，按0.5％和10％比例分别添加含0．4％莫纳可林K红曲于各组日粮内，饲喂22d，观测其对鸡蛋胆固醇变化的影响。结果表明，投药后5—6d，10％添加组和5％添加组蛋黄胆固醇开始下降；15—22d的胆固醇含量与1～5d相比，10％添加组和5％添加组分别下降了9．02％和5．10％，而对照组升高了5．22％。