The complete amino acid sequence of human complex-forming glycoprotein heterogeneous in charge (protein HC) from one individual

The complete amino acid sequence of the single polypeptide chain of human complex-forming glycoprotein heterogeneous in charge (protein HC) isolated from a single individual is reported with the supporting data. The primary structure was determined by automatic degradation of the intact chain and of fragments obtained by chemical and enzymatic degradations of the native or reduced and S-carboxymethylated protein. The polypeptide chain of protein HC contained 182 amino acid residues with a calculated molecular weight of 20,621. No amino acid sequence variability was found and such variability can therefore not explain the great charge heterogeneity of protein HC in a single individual. The amino acid sequence of protein HC was nearly identical to the one reported for human alpha 1-microglobulin in a research communication but contained 15 additional residues.     

The use of cetyltrimethylammonium bromide for the rapid isolation from Staphylococcus aureus of relaxable and non-relaxable plasmid DNA suitable for in vitro manipulation

Plasmid DNA was isolated from Staphylococcus aureus by a rapid method that depends on the precipitation of DNA from cleared lysates by cetyltrimethylammonium bromide at low salt concentrations. The method was validated by its ability to provide DNA for restriction analysis of a highly relaxable plasmid species that is not isolated by more traditional techniques. The DNA can be digested with restriction endonucleases and used for transformation without further purification. The method also provides the basis for analysing staphylococcal plasmids that display a high frequency of deletion after transfer. Simple modifications of the technique allow plasmid DNA to be isolated from other bacteria and the rapid purification of DNA samples before in vitro manipulation.     

Expression of protein HC on the plasma membrane of different human cell types.

The surface expression of a recently described plasma glycoprotein called human complex-forming glycoprotein, hetergeneous in charge (protein HC) on a number of different human cell types was investigated. By means of direct and indirect immunofluorescence, protein HC was shown to be associated with the surface of virtually all cells of the investigated normal cell types including erythrocytes, peripheral blood B and T lymphocytes, and the human fibroblast lines HE 81, HE 31, and WI 38. When transformed and malignant cell populations were studied, it was found that some populations (e.g., the T cell line Molt-4) carried protein HC on the surfaces of very few cells, whereas other cell populations (e.g., chronic lymphocytic leukemia lymphocytes) carried the protein on most cells. Malignant cell populations with intermediary percentages of protein HC-positive cells were also found. Protein HC on the cell surface of normal peripheral blood lymphocytes could be redistributed by incubation of the cells with anti-protein HC-antiserum at 37 degrees C, and this reaction could be inhibited by sodium azide.     

The use of whole barley diets fortified with solutions of urea,minerals and vitamins for lambs

For one experiment 45 early-weaned lambs were given one of the following five diets from weaning to slaughter: (1) whole barley with urea, minerals and vitamins added as a concentrated solution; (2) as diet (1) plus 4 g/kg of sodium sulphate in solution; (3) as diet (2) plus 1.2 g of methionine-hydroxyanalogue (MHA)/kg; (4) as diet (2) plus 2.5 ml of 40% formaldehyde added per kg; (5) a control diet containing whole barley and 100 g/kg of a pelleted supplement based on fish meal. Growth rates (g/d) for the five treatments were 218, 253, 253, 256 and 292. Addition of sulphate significantly increased growth rate and food utilization while MHA had no effect; formalin treatment reduced digestibility and food utilization.In a second experiment 58 lambs were used to study the effect of protein supplements for lambs weaned at various ages and weights. Diets similar to (2) and (5) from Expt. 1 were used, while an intermediate diet (6) was made from an equal mixture of diets (2) and (5). As weaning age increased and as live weight at weaning increased, the difference in growth rate and food utilization between lambs receiving diet (2) and those receiving diets (5) and (6) decreased.It is suggested that for most sheep production systems in which concentrates are used either as the sole feed or as supplements, simple fortification of whole grain with the necessary nutrients is all that is required to achieve optimum results.     

Utilization of alkali-treated grain. 1. Alkali-treated grain in complete diets for steers and lambs

In one experiment, 32 lambs were offered (ad libitum) diets based on whole or NaOH-treated barley, or whole or NaOH-treated oats. The live-weight gains from 17 to 35 kg live weight were 371, 290, 258 and 229 g/day, respectively, while the calculated carcass weight gains were 190, 161, 86 and 118 g/day, respectively. There were large differences in the gut contents at slaughter which were 5.0, 3.3, 9.2 and 3.0 kg for the four diets, respectively.Forty-eight, Hereford-cross steers were allocated to six dietary treatments which consisted of unrestricted access to one of the following diets: NaOH-treated barley; rolled barley; NaOH-treated oats; rolled oats; NaOH-treated wheat; or NaOH-treated maize. The steers were fattened from 325 to 425 kg live weight. The average live weight gains were 1.56, 1.91, 1.37, 1.38, 1.49 and 1.42 kg/day, respectively. The steers receiving rolled barley achieved the greatest live weight gains (P < 0.05). The rumen contents at slaughter were 19.8, 32.2, 24.2, 46.0, 20.8 and 18.4 kg, respectively. Because of differences in gut contents, the only significant differences in carcass weight gain were found for the steers receiving rolled oats, for which carcass gain was lower than that for any other treatment (P < 0.001).Sixteen, Hereford-cross steers were allowed free access to diets containing either NaOH-treated barley alone, or NaOH-treated barley mixed with 10% of chopped straw. The live weight gains from 325 to 425 kg live weight were 1.55 and 1.07 kg/day, and food conversion was 5.7 and 8.4 kg feed/kg gain respectively.     

Demonstration of sequence variations in the promoter region of the human cystatin C gene.

Four point mutations in the promoter region of the human cystatin C gene have been detected by direct sequencing of polymerase chain reaction (PCR) amplified DNA. The four base changes are all localized within a short segment of 85 base pairs. Three cystatin C gene alleles could be defined with respect to these promoter mutations; one with the sequence previously published, one carrying three of the mutations and one with all four base substitutions. Two of the observed mutations are involved in a novel Sst II polymorphism and another generates a new Dde I restriction site. A PCR-based assay for analysis of these Sst II and Dde I sites was designed and used to demonstrate Mendelian inheritance of the polymorphisms as well as to determine the frequencies of the cystatin C gene alleles in the population.