Human lymphocyte tubulin: Purification and characterization in normal and leukemic cells

Tubulin has been purified from human blood and tonsil lymphocytes. Using gel filtration, the molecular weight of human lymphocyte tubulin was estimated to be 119 000. The proteins was shown to consist of two subunits, with molecular weights of 61 000 and 58 000 comparable to the α and β polypeptides of human brain tubulin. A partial identity reaction was observed between lymphocyte tubulin and human tubulin when tested by double immunodiffusion against a rabbit anti-human brain tubulin antibody. In the presence of GTP, the purified protein polymerized to form microtubules. Tubulin was localized to the cell's juxtacentriolar region by immunofluorescence and electron microscopy. When assayed by a colchicine-binding assay corrected for time decay, the binding affinity was 1.50 ± 0.86 · 10⁶M^?1 and a level in normal lymphocytes of 1.21 · 10^?2 ± 0.79 g/g of soluble protein was determined. Since chronic lymphocytic leukemia lymphocytes have an anomalous capping behavior as well as an unusual susceptibility to colchicine toxicity, the properties and levels of tubulin were determined in these cells. Similar values were obtained for the level, decay rate, molecular weight, and K_a for colchicine as for normal lymphocytes. Chronic lymphocytic leukemia lymphocyte tubulin polymerized in a normal fashion. It thus appears that a decrease in the quantity or function of tubulin does not account for these anomalies in the chronic lymphocytic leukemia lymphocyte.     

Introgressive hybridization of tilapias in Zimbabwe

Three species of tilapia native to Zimbabwe, Oreochromis mossambicus, O. mortimeri and O. macrochir are not naturally sympatric, but because of transplantation they have been brought into sympatry. Allozymes of fish examined from Lakes Kariba, Kyle and Chivero showed evidence of introgressive hybridization. The data showed non-random mating populations in Lakes Kariba and Kyle. All three reservoirs displayed individuals with intermediate principal component scores and hybrid index scores indicating hybridization.     

Developmental plasticity in the queen-polymorphic ant Temnothorax longispinosus

Females of social Hymenoptera show developmental plasticity in response to varying social and environmental conditions, though some species have strong genetic influences on the form of the female reproductives. In ants, a queen polymorphism can occur in which large queens initiate new colonies on their own, while small queens enter established nests. Most queen polymorphisms studied to date originate due to genetic differences between individuals of differing form. Here, we report on the development of female form in response to social factors within the nest in the queen-polymorphic ant Temnothorax longispinosus. Three queen size morphs occur: a rare large queen with higher fat stores that can found new colonies independently, a large queen that has low fat stores and is behaviorally flexible, and a small queen that rejoins the natal nest. Both in nesting units collected from the field and those reared in the lab, queen presence during larval development led to fewer larvae developing as gynes (virgin, winged queens), and most of those gynes were the small morph. This queen effect is transferred to developing gyne larvae by close, physical interaction between queens and workers, and causes slower larval development. We conclude that gyne size, and therefore reproductive behavior, in T. longispinosus is developmentally plastic in response to queen presence. Plasticity in reproductive behavior may be an adaptive response to the nest sites utilized by this species. T. longispinosus nests predominantly in acorns and hickory nuts, which can vary dramatically from 1 year to the next. Since queens are more likely to be present in each nesting unit when fewer nest sites are available, the queen effect that results in more small gynes produced links the expression of colony-founding traits to ecological conditions across habitat patches.     

Odor psychophysics and sensory engineering

This paper illustrates the utilization of psychophysical scalingand optimization technology to accomplish two goals. First,by means of ‘goal programming’ the odor scientistcan interrelate chemicals and odor qualities, specify a desiredprofile of odor qualities and determine that combination ofodorants which come as close as possible to generating thatdesired profile. Second, by means of non-linear optimization,the scientist can determine the relation between chemical concentrationsand liking, and furthermore ascertain that combination of chemicalconcentrations which generate maximal liking ratings, subjectto constraints. Constraints include chemical concentrationslying within prespecified limits, and sensory perceptions lyingwithin specific bounds. Mixtures of butyric acid and pulegoneillustrate the approach.     

Purification and characterization of chick intestine brush border membrane Effects of 1α(OH) vitamin D3 treatment

A new technique has been developed for the isolation of membrane vesicles from the vitamin D-deficient and vitamin D-treated chick intestinal brush border membrane. The technique involves removal of nuclei from a low speed pellet by discontinuous sucrose gradient centrifugation. The resulting intact brush borders are then homogenized in 0.5 M Tris and the membrane fragments purified on a glycerol gradient. This preparation represents a 20-fold purification of the brush border marker sucrase. After 1α-hydroxyvitamin D₃ treatment there is a significant increase in membrane phospholipid phosphorous, an alteration in the fatty acid composition of the phosphatidylcholine fraction of membrane phospholipid, and a decrease in sucrase specific activity.     

Methods of tracing hydrogen in polyketide biosynthesis: High field NMR spectroscopy of citrinin produced in a D2O based medium

Penicillium citrinum cultures have been germinated on an H₂O-based medium, resuspended on a D₂O-based medium and treated with [l,2-¹³C₂] acetate. The resulting citrinin (1) has been analysed by²H and¹³C nuclear magnetic resonance spectroscopy and information about the metabolism of hydrogen in citrinin biosynthesis has been deduced.     

Extracellular release of colicin A is non-specific.

The possible involvement of topogenic export sequences within the colicin A polypeptide chain has been investigated. Different constructs have been made using various techniques to introduce deletions in the central and NH2-terminal regions of colicin A. Together, these deletions span the region from amino acid 15 to the end of the protein. None of these regions was found to be required for extracellular release or had any effect on the efficiency of this process. By inserting a termination codon, a Shine-Dalgarno sequence and an initiation codon into the gene for colicin A, the NH2-terminal and central plus COOH-terminal domains could be demonstrated to be released to the same extent when produced as separate polypeptides as when produced as linked ones. The introduction into the COOH-terminal domain of mutations promoting cytoplasmic aggregation had no effect on the secretion of the NH2-terminal polypeptide. These results demonstrated that no specific interaction between the NH2- and COOH-terminal regions of the colicin A polypeptide chain is involved in the release of colicin A. We are led to conclude that there is no topogenic export signal in the polypeptide chain of colicin A involved in the release mechanism. Thus the process is non-specific with respect to the colicin itself and depends solely on the expression of the colicin A lysis protein (Cavard et al., 1985, 1987). The expression of the protein causes the release of not only the colicin but also many other cellular proteins, including beta-lactamase, EF-Tu, and chloramphenicol acetyltransferase.