DNA polymorphisms of the apolipoprotein AII and AI-CIII-AIV genes: a study in men selected for differences in high-density-lipoprotein cholesterol concentration.

We have investigated the frequencies of RFLPs of the apolipoprotein (apo) AII gene and of the apo AI-CIII-AIV gene cluster in 109 men, selected from a random sample of 1,910 men aged 45-59 years, to cover a wide range of plasma high-density-lipoprotein (HDL)-cholesterol concentration. There was no significant difference in apo AI or apo AII RFLP allele frequency between groups of individuals with high and low HDL-cholesterol concentration. However, the apo AI PstI RFLP showed an association with genetic variation determining the plasma concentration of apo AI in this sample. Genetic variation in the apo AI-CIII-AIV gene region, as defined by haplotypes, accounted for 16% of the phenotypic variance in the apo AI concentration and for 8% of the phenotypic variance in HDL-cholesterol concentration. There was no significant association between alleles of the apo AII MspI RFLP and genetic variation determining apo AII or HDL concentration. The data demonstrate that genetic variation in the apo AI-CIII-AIV gene cluster is involved in determining the serum concentration of apo AI in this sample of clinically well individuals.     

A Cladistic Analysis of Phenotype Associations with Haplotypes Inferred from Restriction Endonuclease Mapping. II. the Analysis of Natural Populations

Genes that code for products involved in the physiology of a phenotype are logical candidates for explaining interindividual variation in that phenotype. We present a methodology for discovering associations between genetic variation at such candidate loci (assayed through restriction endonuclease mapping) with phenotypic variation at the population level. We confine our analyses to DNA regions in which recombination is very rare. In this case, the genetic variation at the candiate locus can be organized into a cladogram that represents the evolutionary relationships between the observed haplotypes. Any mutation causing a significant phenotypic effect should be imbedded within the same historical structure defined by the cladogram. We showed, in the first paper of this series, how to use the cladogram to define a nested analysis of variance (NANOVA) that was very efficient at detecting and localizing phenotypically important mutations. However, the NANOVA of haplotype effects could only be applied to populations of homozygous genotypes. In this paper, we apply the quantitative genetic concept of average excess to evaluate the phenotypic effect of a haplotype or group of haplotypes stratified and contrasted according to the nested design defined by the cladogram. We also show how a permutational procedure can be used to make statistical inferences about the nested average excess values in populations containing heterozygous as well as homozygous genotypes. We provide two worked examples that investigate associations between genetic variation at or near the Alcohol dehydrogenase (Adh) locus and Adh activity in Drosophila melanogaster, and associations between genetic variation at or near some apolipoprotein loci and various lipid phenotypes in a human population.     

A fermentor for study of sauerkraut fermentation

A fermentor was designed and constructed for study of the physical, microbiological, and chemical changes that occur during the sauerkraut fermentation. The fermentor has some essential features that include restriction in volume of the sauerkraut bed, construction of clear plastic to permit visual determination of liquid-level changes as a result of gas entrapment within the sauerkraut bed, and a gas-lift device for use in nitrogen purging of the fermenting brine. Fermentations exhibited two distinct stages, the first one gaseous and the second non-gaseous. The gaseous stage was characterized by rapid CO(2) and acid production due to growth by hetero-fermentative lactic acid bacteria with resultant gas entrapment within the sauerkraut bed and a rise in liquid level. Also, rapid disappearance of fructose and rapid appearance of mannitol occurred during this stage. The nongaseous stage was characterized by growth of homo-fermentative lactic acid bacteria with little or no CO(2) production and a gradual increase in lactic acid until all fermentable sugars were metabolized. Nitrogen purging appeared to offer several potential advantages, including a means for brine circulation, removal of CO(2) from the brine, and anaerobiosis to ensure retention of ascorbic acid, desirable color, and other oxygen-sensitive traits in sauerkraut.     

Systematics as science: A response to Cronquist

Our reply to the commentary on cladistics presented by Cronquist (1987) is aimed at four issues:

1. the application of scientific principles in systematics;
2. the recognition that the analysis of pattern is a vital precursor to any consideration of evolutionary process. A priori judgements of evolutionary process are unnecessary for the generation of informative systematic hypotheses which are chosen for their ability to explain the patterns of character distributions rather than for compatibility with any particular preconceived ideas about evolution;
3. that phenetic concepts such as overall similarity, grades, gaps, and degree of divergence, if included in methods of phylogenetic inference, will give erroneous results. Paraphyletic and polyphyletic groups must, consequently, be rejected from systematics since they have no rational empirical basis for recognition;
4. the fact that many of the problems of phylogenetic analysis attributed by Cronquist to cladistics are common to all systematic methods but that these can be dealt with by the application of such principles as parsimony, synapomorphy, and strict monophyly.

     

Collagen can modulate cell interactions with fibronectin

We have examined the effects of soluble collagen on the function of fibronectin in baby hamster kidney (BHK) cells. Collagen and its purified alpha1(l) chain noncompetitively inhibited cell spreading on substrates precoated with fibronectin or a 75,000-D cell-binding fragment of fibronectin. Neither preincubation of cells with collagen followed by washing nor the addition of collagen to previously spread cells had any inhibitory effect on cell spreading, which indicates a requirement for the concurrent presence of collagen during the process of spreading. Treatment of collagen or alpha1(l) chain with collagenase abolished the inhibitory effect on fibronectin-mediated cell spreading. However, direct attachment of BHK cells to fibronectin-coated or 75,000-D fragment-coated substrates was not inhibited by collagen or by the alpha1(l) chain. Moreover, the binding of [3H]fibronectin or the 3'-75,000-D fragment to cell surfaces was not inhibited by the presence of soluble collagen, whereas soluble fibronectin inhibited binding. Although the binding of [3H]fibronectin-coated beads to BHK cell surfaces was also not inhibited by collagen, the phagocytosis of such beads was inhibited by the presence of collagen. On the other hand, soluble fibronectin partially inhibited the binding of fibronectin-coated beads but did not inhibit phagocytosis of the beads that did bind. The mechanism of the inhibition of fibronectin function by collagen and the possible interactions of two different kinds of receptors on the cell surface are discussed.     

Identification of a deletion in the low density lipoprotein (LDL) receptor gene in a patient with familial hypercholesterolaemia

Summary DNA samples from 60 unrelated UK patients with familial hypercholesterolaemia (FH) were screened by Southern blot hybridisation to detect gross alterations in the low density lipoprotein (LDL) receptor gene. One patient was found to have a 2kb deletion in the 3 part of the gene. The deletion cosegregates with the FH phenotype in his family. This finding is compatible with the deletion being the cause of FH in this case and makes a presymptomatic test based on DNA analysis available for this family. The defects in most of the other patients are likely to be due to point mutations.     

Cholesterol-free phospholipid domains may be the membrane feature selected by N epsilon-dansyl-L-lysine and merocyanine 540

We have used N epsilon-dansyl-L-lysine as a fluorescent membrane probe, to study cells taken from tissues concerned with immune function. There is a striking similarity between the staining selectivity of this compound and that reported by others for merocyanine 540. Both compounds stain leukemic, human, peripheral leukocytes, an erythroleukemia line, and some mouse bone marrow cells, suggesting common selectivity for a membrane feature of hemopoietic cells. Both compounds fail to stain red blood cells, normal human leukocytes, mouse spleen and thymus cells. We have recently reported that dansyl-lysine apparently selects for cholesterol-free phospholipid domains in liposomes and now report similar selectivity for merocyanine 540 staining of liposomes.     

Dansyl lysine: a structure-selective fluorescent membrane stain?

Dansyl lysine (DL) is a fluorescent compound that has significantly higher solubility in synthetic phosphatidylcholine (PC) membranes with a low cholesterol content than it does in water or in membranes having a high cholesterol content. Its fluorescence intensity is enhanced at least 50-fold when dissolved in PC membranes. Therefore, membranes with mole fractions of cholesterol (Xch) less than or equal to 0.5-0.3 are stained by aqueous solutions of DL: those with a higher cholesterol content, 0.3-0.4 less than or equal to Xch less than or equal to 0.5, are not. It is proposed that DL selects for a structural feature of membranes: cholesterol-free domains. The phenomenon has provided evidence for long-lived compositional heterogeneity in large multilamellar PC-cholesterol liposomes having Xch less than or equal to 0.2. This is not consistent with a model in which the homogeneous state is thermodynamically favored and both intermembrane transfer and transmembrane transfer (flip-flop) of cholesterol are fast. These studies are of potential importance for understanding cell membrane structure, in particular lipid-phase equilibria and the maintenance of compositional heterogeneity between the different membranes of cells.     

Photosynthesis in sugar beet depends on root growth

Summary Sugar-beet plants, germinated in growth cabinets at 20°C and transplanted into the field after 3 weeks, developed much larger roots than plants grown from seed drilled directly into the soil. At the end of the season, the roots of transplants were 39% greater than from drilled seed—an increase of 14 m tons per hectare. The increased yield was mainly due to a sustained increase in photosynthesis because of the larger sink for carbohydrates provided by plants from the growth cabinets.