We report a new method for in situ localization of DNA sequences that allows excellent preservation of nuclear and chromosomal ultrastructure and direct, in vivo observations. 256 direct repeats of the lac operator were added to vector constructs used for transfection and served as a tag for labeling by lac repressor. This system was first characterized by visualization of chromosome homogeneously staining regions (HSRs) produced by gene amplification using a dihydrofolate reductase (DHFR) expression vector with methotrexate selection. Using electron microscopy, most HSRs showed approximately 100-nm fibers, as described previously for the bulk, large-scale chromatin organization in these cells, and by light microscopy, distinct, large-scale chromatin fibers could be traced in vivo up to 5 microns in length. Subsequent experiments demonstrated the potential for more general applications of this labeling technology. Single and multiple copies of the integrated vector could be detected in living CHO cells before gene amplification, and detection of a single 256 lac operator repeat and its stability during mitosis was demonstrated by its targeted insertion into budding yeast cells by homologous recombination. In both CHO cells and yeast, use of the green fluorescent protein-lac repressor protein allowed extended, in vivo observations of the operator-tagged chromosomal DNA. Future applications of this technology should facilitate structural, functional, and genetic analysis of chromatin organization, chromosome dynamics, and nuclear architecture.

Many developing insect neurones pass through a phase when they respond to nitric oxide (NO) by producing cyclic GMP. Studies on identified grasshopper motoneurones show that this NO sensitivity appears after the growth cone has arrived at its target but before it has started to send out branches. NO sensitivity typically ends as synaptogenesis is nearing completion. Data from interneurones and sensory neurones are also consistent with the hypothesis that NO sensitivity appears as a developing neurone changes from axonal outgrowth to maturation and synaptogenesis. Cyclic GMP likely constitutes part of a retrograde signalling pathway between a neurone and its synaptic partner. NO sensitivity also appears in some mature neurones at times when they may be undergoing synaptic rearrangement. Comparative studies on other insects indicate that the association between an NO-sensitive guanylate cyclase and synaptogenesis is an ancient one, as evidenced by its presence in both ancient and more recently evolved insect groups.

In species where males and females differ in number of sex chromosomes, the expression of sex-linked genes is equalized by a process known as dosage compensation. In Drosophila melanogaster, dosage compensation is mediated by the binding of the products of the male-specific lethal (msl) genes to the single male X chromosome. Here we report that the sex- and chromosome-specific binding of three of the msl proteins (MSLs) occurs in other drosophilid species, spanning four genera. Moreover, we show that MSL binding correlates with the evolution of the sex chromosomes: in species that have acquired a second X chromosome arm because of an X-autosome translocation, we observe binding of the MSLs to the 'new' (previously autosomal) arm of the X chromosome, only when its homologue has degenerated. Moreover, in Drosophila miranda, a Y-autosome translocation has produced a new X chromosome (called neo-X), only some regions of which are dosage compensated. In this neo-X chromosome, the pattern of MSL binding correlates with the known pattern of dosage compensation.

A formalism is given in which the optical field generated by a near-field optical aperture is described as an analytic expansion over a complete set of optical modes. This vectoral solution preserves the divergent behavior of the near field and the dipolar nature of the far field. Numerical calculation of the fields requires only evaluation of a well behaved, one-dimensional integral. The formalism is directly applicable to experiments in near-field scanning optical microscopy when relatively flat samples are evaluated.

Colonies of the aphidPseudoregma alexanderi produce morphologically-specialized first-instar larvae, termed soldiers, that defend the colony from predators. The environmental cues and physiological mechanisms governing soldier production are currently unknown. Here we present a morphometric study of soldiers and normal first-instar larvae ofP. alexanderi. Several morphological features (fore-leg length and width, hind-leg length, and horn length) plotted against body length display relationship that are similar to a sigmoidal curve. We found further support for an earlier finding that soldiers fall into two size categories, majors and minors, although both types of soldiers appear to follow the same allometry. The patterns of allometry in the soldier-producing aphids are very different from those found in other social insects and do not easily fit into the traditional categorization of allometries. We present two simple alternative models of soldier development as a framework for guiding future studies of the mechanisms of soldier production.

We report the experimental studies of a parametric excitation of a second sound (SS) by a first sound (FS) in a superfluid helium in a resonance cavity. The results on several topics in this system are presented: (i) The linear properties of the instability, namely, the threshold, its temperature and geometrical dependencies, and the spectra of SS just above the onset were measured. They were found to be in a good quantitative agreement with the theory. (ii) It was shown that the mechanism of SS amplitude saturation is due to the nonlinear attenuation of SS via three wave interactions between the SS waves. Strong low frequency amplitude fluctuations of SS above the threshold were observed. The spectra of these fluctuations had a universal shape with exponentially decaying tails. Furthermore, the spectral width grew continuously with the FS amplitude. The role of three and four wave interactions are discussed with respect to the nonlinear SS behavior. The first evidence of Gaussian statistics of the wave amplitudes for the parametrically generated wave ensemble was obtained. (iii) The experiments on simultaneous pumping of the FS and independent SS waves revealed new effects. Below the instability threshold, the SS phase conjugation as a result of three-wave interactions between the FS and SS waves was observed. Above the threshold two new effects were found: a giant amplification of the SS wave intensity and strong resonance oscillations of the SS wave amplitude as a function of the FS amplitude. Qualitative explanations of these effects are suggested.

1. Defensive individuals, termed soldiers, have recently been discovered in aphids, Soldiers are typically early instar larvae, and in many species the soldiers are reproductively sterile and morphologically and behaviourally specialized. 2. Since aphids reproduce parthenogenetically, we might expect soldier production to be more widespread in aphids than it is. We suggest that a more useful way to think about these problems is to attempt to understand how a clone (rather than an individual) should invest in defence and reproduction. 3. Known soldiers are currently restricted to two families of aphids, the Pemphigidae and Hormaphididae, although they are distributed widely among genera within these families. We discuss the use of a phylogenetic perspective to aid comparative studies of soldier production and we demonstrate this approach using current estimates of phylogenetic affinities among aphids. We show that the distribution of soldier production requires a minimum of six to nine evolutionary origins plus at least one loss. 4. At least four main types of soldiers exist and we present and discuss this diversity of soldiers. 5. Most soldier-producing species produce soldiers within plant galls and we discuss the importance of galls for the evolution of soldiers. 6. We summarize the evidence on the interactions between soldiers and predators and between soldier-producing aphids and ants. 7. We present an optimality model for soldier investment strategies to help guide investigations of the ecological factors selecting for soldiers. 8. The proximate mechanisms of soldier production are currently very poorly understood and we suggest several avenues for further research.

We reanalysed Yang & Pattern's allozyme data, published in Auk in 1981, of Darwin's finches with a variety of distance and cladistic methods to estimate the phylogeny of the group. Different methods yielded different results, nevertheless there was widespread agreement among the distance methods on several groupings. First, the two species of Camarhynchus grouped near one another, but not always as a monophyletic group. Second, Cactospiza pallida and Platyspiza crassirostris formed a monophyletic group. Finally, all the methods (including parsimony) supported the monophyly of the ground finches. The three distance methods also found close relationships generally between each of two populations of Geospiza scandens, G. difficilis and G. conirostris. There is evidence for inconstancy of evolutionary rates among species. Results from distance methods allowing for rate variation among lineages suggest three conclusions which differ from Yang and Patton's findings. First, the monophyletic ground finches arose from the paraphyletic tree finches. Yang and Patton found that the ground finches and tree finches were sister monophyletic taxa. Second, Geospiza scandens appears to be a recently derived species, and not the most basal ground finch. Third, G. fuliginosa is not a recently derived species of ground finch, but was derived from an older split from the remaining ground finches. Most of these conclusions should be considered tentative both because the parsimony trees disagreed sharply with the distance trees and because no clades were strongly supported by the results of bootstrapping and statistical tests of alternative hypotheses. Absence of strong support for clades was probably due to insufficient data. Future phylogenetic studies, preferably using DNA sequence data from several unlinked loci, should sample several populations of each species, and should attempt to assess the importance of hybridization in species phylogeny.

Apoptotic cell death is a mechanism by which organisms eliminate superfluous or harmful cells. Expression of the cell death regulatory protein REAPER (RPR) in the developing Drosophila eye results in a small eye owing to excess cell death. We show that mutations in thread (th) are dominant enhancers of RPR-induced cell death and that th encodes a protein homologous to baculovirus inhibitors of apoptosis (IAPs), which we call Drosophila IAP1 (DIAP1). Overexpression of DIAP1 or a related protein, DIAP2, in the eye suppresses normally occurring cell death as well as death due to overexpression of rpr or head involution defective. IAP death-preventing activity localizes to the N-terminal baculovirus IAP repeats, a motif found in both viral and cellular proteins associated with death prevention.

Selection for mutants which release glucose repression of the CYB2 gene was used to identify genes which regulate repression of mitochondrial biogenesis. We have identified two of these as the previously described GRR1/CAT80 and ROX3 genes. Mutations in these genes not only release glucose repression of CYB2 but also generally release respiration of the mutants from glucose repression. In addition, both mutants are partially defective in CYB2 expression when grown on nonfermentable carbon sources, indicating a positive regulatory role as well. ROX3 was cloned by complementation of a glucose-inducible flocculating phenotype of an amber mutant and has been mapped as a new leftmost marker on chromosome 2. The ROX3 mutant has only a modest defect in glucose repression of GAL1 but is substantially compromised in galactose induction of GAL1 expression. This mutant also has increased SUC2 expression on nonrepressing carbon sources. We have also characterized the regulation of CYB2 in strains carrying null mutation in two other glucose repression genes, HXK2 and SSN6, and show that HXK2 is a negative regulator of CYB2, whereas SSN6 appears to be a positive effector of CYB2 expression.