Dolichyl phosphate phosphatase from Tetrahymena pyriformis.

A soluble dolichyl phosphate phosphatase from Tetrahymena pyriformis was purified about 68-fold. The enzyme appeared to be specific for dolichyl phosphate and existed in two interrelated forms, one of mol.wt. about 500000 and the other of mol.wt. about 63000. The enzyme was strongly inhibited by 5 mM-Mn2+ and was strongly stimulated by Mg2+. Tetrahymena in the exponential growth phase contained more of this enzymic activity than cells in stationary or lag phase. The dolichyl phosphate phosphatase may be loosely bound to mitochondrial membranes. Two roles proposed for this enzyme are (1) that of releasing dolichol from its phosphorylated biosynthetic form for its use in the cell as unesterified dolichol or dolichyl ester and/or (2) that of regulation of synthesis of glycoproteins or some other glycosylated compound.     

Inducible α-glucosidase of Mucor rouxii: Effects of dimorphism on the development of the wall-bound activity

Some properties of the inducible α-glucosidase system of Mucor rouxii were investigated. This enzymatic activity was induced after resuspending glucose-grown cells in a maltose-supplemented medium. The wall-bound activity of α-glucosidase was determined by using intact cells in the enzymatic assay; this activity represented from 80 to 90% of the total activity present in the induced cells. The addition of glucose before, or during, the induction period repressed α-glucosidase synthesis. α-Glucosidase induction was tested under aerobic and anaerobic conditions. It was found that the enzyme synthesis and the appearance of wall-bound activity were not affected by changing the gaseous environment. On the other hand, it was observed that anaerobically grown yeast-like cells were much less efficient than aerobic mycelia to develop wall-bound α-glucosidase activity. This could explain earlier observations about the incapacity of M. rouxii to utilize maltose as a substrate for anaerobic growth. This idea was strengthened by the fact that, if an anaerobic culture was induced to develop under a mycelial morphology by adding to the medium the chemical agent EDTA, these cells also acquired the capacity to grow on maltose and concomitantly possessed wall-bound α-glucosidase activity. The relevance of the structure of the cell wall on the capacity of M. rouxii to metabolize maltose is discussed.     

Binomial sequential sampling of adult Saccharicoccus sacchari on sugarcane

The between-stalk dispersion characteristics of adults of the pink sugarcane mealybug Saccharicoccus sacchari (Cockerell) were determined in southern Queensland. Iwao's patchiness regression was inappropriate to describe the relationship between mean and variance. Taylor's power law indicated that adults were aggregated, especially at the beginning and end of the ratoon growth period. Binomial data were modelled by the Nachman model; the model of Grout and two models of Wilson & Room were inappropriate to describe the relationship between proportion of stalks infested and mean numbers of adults per stalk. Relationships to determine sample sizes for fixed levels of precision and binomial fixed-precision-level stop lines are developed for different sampling times using Taylor's power law and Nachman's equation.     

Our Planet,Our Health

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Our Planet, Our Health Report of the who commission on health and environment     

A simple method of generating axenic derivatives of Dictyostelium strains

We describe a generally applicable method of adapting Dictyostelium from growth on a bacterial food source to axenic growth. Cells are initially selected by growth on a plastic substratum but subsequently acquire the ability to grow in suspension culture. These strains can be transformed efficiently by DNA-mediated gene transfer.     

Spatial distribution and sequential sampling of adults ofGonocephalum macleayi andPterohelaeus darlingensis in different cropping regimes

Counts of adults of the false wirewormsGonocephalum macleayi (Blackburn) andPterohelaeus darlingensis Carter in pitfall traps in burnt, mulched and sorghum treatments conformed to Taylor's power law. Within a species there were no significant differences in distributions of counts of either sex in any habitat butG. macleayi were more aggregated thanP. darlingensis (Taylor'sb 1.35 and 1.26, respectively). Relationships to determine sample sizes for fixed levels of precision and fixed-precision-level stop lines for sequential sampling are developed for each species using Taylor's parameters for combined data over all habitats.     

Telomere end-replication problem and cell aging.

Since DNA polymerase requires a labile primer to initiate unidirectional 5'-3' synthesis, some bases at the 3' end of each template strand are not copied unless special mechanisms bypass this "end-replication" problem. Immortal eukaryotic cells, including transformed human cells, apparently use telomerase, an enzyme that elongates telomeres, to overcome incomplete end-replication. However, telomerase has not been detected in normal somatic cells, and these cells lose telomeres with age. Therefore, to better understand the consequences of incomplete replication, we modeled this process for a population of dividing cells. The analysis suggests four things. First, if single-stranded overhangs generated by incomplete replication are not degraded, then mean telomere length decreases by 0.25 of a deletion event per generation. If overhangs are degraded, the rate doubles. Data showing a decrease of about 50 base-pairs per generation in fibroblasts suggest that a full deletion event is 100 to 200 base-pairs. Second, if cells senesce after 80 doublings in vitro, mean telomere length decreases about 4000 base-pairs, but one or more telomeres in each cell will lose significantly more telomeric DNA. A checkpoint for regulation of cell growth may be signalled at that point. Third, variation in telomere length predicted by the model is consistent with the abrupt decline in dividing cells at senescence. Finally, variation in length of terminal restriction fragments is not fully explained by incomplete replication, suggesting significant interchromosomal variation in the length of telomeric or subtelomeric repeats. This analysis, together with assumptions allowing dominance of telomerase inactivation, suggests that telomere loss could explain cell cycle exit in human fibroblasts.     

Density dependent interactions between VA mycorrhizal fungi and even-aged seedlings of two perennial Fabaceae species

Summary The interaction of density and mycorrhizal effects on the growth, mineral nutrition and size distribution of seedlings of two perennial members of the Fabaceae was investigated in pot culture. Seedlings of Otholobium hirtum and Aspalathus linearis were grown at densities of 1, 4, 8 and 16 plants per 13-cm pot with or without vesicular-arbuscular (VA) mycorrhizal inoculum for 120 days. Plant mass, relative growth rates, height and leaf number all decreased with increasing plant density. This was ascribed to the decreasing availability of phosphorus per plant as density increased. O. hirtum was highly dependent on mycorrhizas for P uptake but both mycorrhizal and non-mycorrhizal A. linearis seedlings were able to extract soil P with equal ease. Plant size distribution as measured by the coefficient of variation (CV) of shoot mass was greater at higher densities. CVs of mycorrhizal O. hirtum plants were higher than those of non-mycorrhizal plants. CVs of the facultatively mycorrhizal A. linearis were similar for both mycorrhizal and non-mycorrhizal plants. Higher CVs are attributed to resource preemption by larger individuals. Individuals in populations with high CVs will probably survive stress which would result in the extinction of populations with low CVs. Mass of mycorrhizal plants of both species decreased more rapidly with increasing density than did non-mycorrhizal plant mass. It is concluded that the cost of being mycorrhizal increases as plant density increases, while the benefit decreases. The results suggest that mycorrhizas will influence density-dependent population processes of faculative and obligate mycorrhizal species.     

Plasma and tissue alterations of peptide YY and enteroglucagon in rats after colectomy.

Peptide YY (PYY) and enteroglucagon are produced by endocrine cells of the colonic mucosa. PYY inhibits upper gastrointestinal motility, and enteroglucagon is trophic for small bowel mucosa. Adaptive increase in the production and release of these peptides may improve functional results after colorectal resections. We hypothesized that if segments of the colon were resected, then production and release of PYY and enteroglucagon would increase in the remaining segments of bowel. Animals which underwent colonic transections and partial resections had transient elevations of PYY up to 250 +/- 80 pmol/L, which dropped to control group levels in the second week following surgery. Rats with an abdominal colectomy had significantly greater PYY levels than all other groups from the third (208 +/- 30 pmol/L) to the thirty-eighth (100 +/- 16 pmol/L) week of the study. Circulating levels of enteroglucagon were elevated to 156 +/- 35 pmol/L in rats with a right hemicolectomy during the first week following surgery. Enteroglucagon levels did not significantly vary in the other groups studied. Both tissue PYY (413 +/- 33 pmol/gram) and tissue enteroglucagon (171 +/- 17 pmol/gram) were significantly elevated in the rectums of the rats with an abdominal colectomy, as compared to all other groups. The elevated tissue levels may thus account for the ability to maintain elevated plasma PYY. Double immunogold labeling of endocrine cells in the colorectal tissue for PYY and enteroglucagon revealed both peptides within the same endocrine cells and secretory granules. These studies support the hypothesis that circulating levels of PYY are elevated after major colonic resections and suggest that L-type endocrine cells may participate in adaptive responses which improve intestinal function following colonic surgery.     

Immunoassay using 125I- or enzyme-labeled protein A and antigen-coated tubes

Antigen-coated plastic tubes were used with ¹²⁵I- or enzyme-labeled staphylococcal protein A in a general immunoassay method for antigens and haptens. Protein A reacts with immunoglobulin G (IgG) regardless of antibody specificity at sites distal to the antigen combining site and does not inhibit the immune reaction. It therefore serves as a general tracer and its use eliminates the need to purify and to label individual components for each assay. Macromolecular antigens were bound to polystyrene or polypropylene tubes by direct passive adsorption. Haptens with free carboxyl groups were bound covalently to poly-l-lysine and these conjugates passively adsorbed to the tube surface. Optimal assay conditions were established for the quantitative determination of immunoglobulins and the folate derivatives, methotrexate and 5-methyltetrahydrofolate, using ¹²⁵I-labeled protein A or protein A labeled with alkaline phosphatase. The method has been used to estimate levels of IgG, IgA, IgM, and IgE in serum in volumes up to 1 ml.