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.     

Comparative virulence of NAD-dependent and NAD-independent Actinobacillus pleuropneumoniae strains.

The virulence of a NAD-independent Actinobacillus pleuropneumoniae serotype 2 strain and NAD-dependent serotype 2, 3 and 9 strains was compared under experimental conditions. Hysterectomy-derived piglets were inoculated endobronchially with 50-500 cfu of these strains. All 23 piglets inoculated with the NAD-dependent strains developed acute disease within 12 hours post inoculation. Twenty-two of these piglets died within 24 hours after the first clinical signs. Three of nine piglets inoculated with the NAD-independent strain did not develop clinical disease. In the other six piglets, disease signs were similar as in the piglets inoculated with the NAD-dependent strains. No differences in clinical disease were observed between colostrum deprived piglets and piglets that obtained colostrum from a SPF sow.     

Surgery of hepatic cysts in a cat.

A 2-year-old female Japanese domestic cat weighing 3.6 kg was presented due to marked abdominal distention and was diagnosed as hepatic cyst. The bile acid concentration of the hepatic cyst was as low as the serum bile acid concentration. The membrane of the cysts was excised from the edge of the compressed hepatic tissue and no other surgical manipulation was performed. This surgical procedure is simple and quick compared to hepatic lobectomy, however, it is necessary to evaluate the fluid of the cyst before performing this procedure.     

Effect of Completion of Vernalization on Generative Development of Winter Rape (Brassica napus L. var. oleifera) Plants

Germinating seeds and young plants of winter rape var. Górczañski were vernalized for 56–63 days under conditions of 9-hour day, at the temperature 2 and 5 °C and in continuous darkness at the temperature 2 °C. After vernalization the plants grew under conditions enabling to complete vernalization: in a glass-house at the temperature day/night 15/10 °C and in semi natural conditions of open vegetation hall in the period from June till August. After sub-optimal vernalization further growth of the plants at lowered temperature increased its effectiveness (completion of vernalization). Depending on the degree of the vernalization of the plants the completion of their vernalization was both obligatory, i.e. conditioning the acquisition of the ability of generative development, and facultative i.e. accelerating this development. It has been demonstrated that the population of plants of the examined variety is strongly differentiated not only with respect of vernalization requirements in the particular plants, but also what regards the effectiveness of vernalization completion. New observations have been made indicating that the mechanisms controlling the successive phases of generative development, i.e. phase of forming flower buds and the flowering phase are not identical which may be interpreted as indicating that the "flowering factor" is polymorphous.     

Analysis of Growth and Productivity of Indian Mustard (Brassica juncea) in Relation to FYM, Nitrogen and Source of Fertilizer

A field experiment was conducted during the winter seasons of 1992–93 and 1993–94 at Anand to study the effect of FYM, nitrogen and source of fertilizer on growth and yield of mustard [ Brassica juncea (L.) Czernj & Cosson]. The results showed significant variation in leaf area index (LAI), crop growth rate (CGR), dry matter production and seed yield. The direct effect of farmyard manure (FYM) was conspicuous in improving the growth of mustard. FYM application at 10 tonnes ha⁻¹ significantly increased the LAI, CGR and dry matter accumulation per plant at almost all the stages during first year study (1992–93) and in pooled analysis. Similarly, nitrogen application registered maximum LAI, CGR at 75 kg level and RGR and NAR at 50 kg level at almost all the during both years. Sulphur carrying source (Ammonium sulphate plus single super phosphate) increased all stages growth characters. Maximum dry matter accumulation per plant and seed yield were recorded with highest levels of FYM (20 tonnes ha⁻¹), N (75 kg ha⁻¹) and source having S. Seed yield was strongly associated with LAI and dry matter accumulation per plant at all the stages.