Identification of a novel Drosophila opsin reveals specific patterning of the R7 and R8 photoreceptor cells.

The function of the compound eye is dependent upon a developmental program that specifies different cell fates and directs the expression of spectrally distinct opsins in different photoreceptor cells. Rh5 is a novel Drosophila opsin gene that encodes a biologically active visual pigment that is expressed in a subset of R8 photoreceptor cells. Rh5 expression in the R8 cell of an individual ommatidium is strictly coordinated with the expression of Rh3, in the overlying R7 cell. In sevenless mutant files, which lack R7 photoreceptor cells, the expression of the Rh5 protein in R8 cells is disrupted, providing evidence for a specific developmental signal between the R7 and R8 cells that is responsible for the paired expression of opsin genes.

Efficient in vivo gene delivery by the negatively charged complexes of cationic liposomes and plasmid DNA.

We examined changes in zeta potential (the surface charge density, zeta) of the complexes of liposome (nmol)/DNA (microg) (L/D) formed in water at three different ratios (L/D=1, 10 and 20) by changing the ionic strength or pH to find an optimum formulation for in vivo gene delivery. At high DNA concentrations, zeta of the complexes formed in water at L/D=10 was significantly lowered by adding NaCl (zeta=+8.44+/-3.1 to -27.6+/-3.5 mV) or increasing pH from 5 (zeta=+15.3+/-1.0) to 9 (zeta=-22.5+/-2.5 mV). However, the positively charged complexes formed at L/D=20 (zeta=+6.2+/-3.5 mV) became negative as NaCl was added at alkaline pH as observed in medium (zeta=-19.7+/-9.9 mV). Thus, the complexes formed in water under the optimum condition were stable and largely negatively charged at L/D=1 (zeta=-58.1+/-3.9 mV), unstable and slightly positively charged at L/D=10 (zeta=+8.44+/-3.7 mV), and unstable and largely positively charged at L/D=20 (zeta=+24.3+/-3.6 mV). The negatively charged complexes efficiently delivered DNA into both solid and ascitic tumor cells. However, the positively charged complexes were very poor in delivering DNA into solid tumors, yet were efficient in delivering DNA into ascitic tumors grown in the peritoneum regardless of complex size. This slightly lower gene transfer efficiency of the negatively charged complexes can be as efficient as the positively charged ones when an injection is repeated (at least two injections), which is the most common case for therapy regimes. The results indicate that optimum in vivo lipofection may depend on the site of tumor growth.

Nitric oxide-mediated tumor cell killing of cisplatin-based interferon-gamma gene therapy in murine ovarian carcinoma.

We have determined the role of nitric oxide (NO)-mediated tumor cell killing in the treatment of an animal model of murine ovarian carcinoma grown in the peritoneum with a combination of cisplatin and cationic liposomes containing an expression vector for interferon-gamma (IFN-gamma). The approach was to determine whether the therapy was effective in mice homozygous for a disrupted inducible NO synthase (iNOS) allele; these mice were unable to produce NO in response to IFN-gamma. iNOS (-/-) mice treated with both cisplatin and liposomal IFN-gamma gene did not produce a significant amount of NO in ascites (12.1+/-4.5 microM), although they expressed a high level of IFN-gamma (9002+/-723 U/mL of ascitic fluid). As a result, mice died of tumors within 11-62 days. However, wild-type mice treated with both cisplatin and liposomal IFN-gamma gene produced a significant amount of NO in ascites (113.7+/-17.9 microM) with a high level of IFN-gamma gene expression (9350+/-1254 U/mL of ascitic fluid) and were free of tumors for at least 80 days. This result confirmed that NO was a direct mediator of IFN-gamma cytotoxicity.

Comparison of pollutant runoff in Lake Biwa tributaries, Japan and the Brunette River watershed, Canada.

This study is aimed at verifying runoff pollutant loadings from urban areas. Urban runoff has been considered an important source of diffuse pollution especially during storm events. This paper describes the pollutant runoff during storm events, mainly in terms of effects of watershed characteristics. Data collected from Lake Biwa tributaries, Japan, have shown fundamental information to control pollutant runoff into receiving water. Also, data from the Brunette River watershed, Canada, which is a highly urbanized watershed in the Vancouver region, have been used for a comparative analysis. In the results, available information for the environmental management of urban storm water runoff was obtained by comparing the data on pollutant runoff in both watersheds.

The conduct of socially sensitive research: sex offenders as participants.

This article examines some extra-research variables inherent in conducting community-based research with child molesters and other sex offenders. These include such issues as informed consent with court-referred participants, confidentiality when interagency collaboration is necessary, and duty to take proper care. Methods for establishing the scientific credibility of the project, responding to client grievances, dealing with client crises, and protecting staff and the surrounding community also are addressed. Adopting procedures similar to those described here could be an important step in ensuring the viability of other socially sensitive research projects.

Stable chromosome fission associated with rDNA mobility.

A spontaneous chromosome fission in the plant Hypochoeris radicata has been characterized by Feulgen staining, in situ hybridization of the rDNA probe pTA71 and silver staining for active nucleolus organizing regions. The parental acrocentric chromosome has no detectable ribosomal genes at the centromere, but both fission derivatives possess active NORs at their centric ends. In fission heterozygotes, pachytene configurations studied by synaptonemal complex spreading show that the ribosomal cistrons form short arms on each telocentric which pair together to form a triradial. The paired short arms are associated with the single nucleolus at pachytene. It is proposed that the origin and stabilization of the fission rearrangement involved transposition of rDNA from the nucleolus organizing region of chromosome 3 into the centromeric region of chromosome 1.

Determination of Poly-beta-Hydroxybutyrate and Poly-beta-Hydroxyvalerate in Activated Sludge by Gas-Liquid Chromatography.

A convenient gas-liquid chromatography procedure to quantify poly-beta-hydroxybutyrate and poly-beta-hydroxyvalerate in activated sludge was developed by combining lyophilization of the samples, purification of the chloroform phase by water reextraction, and the use of capillary columns. With a flame ionization detector the sensitivity was estimated at 10 g/liter.

Purple sulfur bacteria control the growth of aerobic heterotrophic bacterioplankton in a meromictic salt lake.

In meromictic Mahoney Lake, British Columbia, Canada, the heterotrophic bacterial production in the mixolimnion exceeded concomitant primary production by a factor of 7. Bacterial growth rates were correlated neither to primary production nor to the amount of chlorophyll a. Both results indicate an uncoupling of bacteria and phytoplankton. In the chemocline of the lake, an extremely dense population of the purple sulfur bacterium Amoebobacter purpureus is present year round. We investigated whether anoxygenic phototrophs are significant for the growth of aerobic bacterioplankton in the overlaying water. Bacterial growth rates in the mixolimnion were limited by inorganic phosphorus or nitrogen most of the time, and the biomass of heterotrophic bacteria did not increase until, in autumn, 86% of the cells of A. purpureus appeared in the mixolimnion because of their reduced buoyant density. The increase in heterotrophic bacterial biomass, soluble phosphorus concentrations below the detection limit, and an extraordinarily high activity of alkaline phosphatase in the mixolimnion indicate a rapid liberation of organically bound phosphorus from A. purpureus cells accompanied by a simultaneous incorporation into heterotrophic bacterioplankton. High concentrations of allochthonously derived dissolved organic carbon (mean, 60 mg of C(middot)liter(sup-1)) were measured in the lake water. In Mahoney Lake, liberation of phosphorus from upwelling purple sulfur bacteria and degradation of allochthonous dissolved organic carbon as an additional carbon source render heterotrophic bacterial production largely independent of the photosynthesis of phytoplankton. A recycling of inorganic nutrients via phototrophic bacteria also appears to be relevant in other lakes with anoxic bottom waters.

The relationship between genetic and cytogenetic maps of pea. I. Standard and translocation karyotypes.

A detailed cytogenetical study of inbred lines of pea and their F1 hybrids has been undertaken to study the relationship between the cytogenetic map and the molecular linkage map. The mitotic karyotypes of a standard pea line, JI15, a translocation line, JI61, and line JI281, a line used in the production of a mapping population, are given. A chromosome rearrangement detected by cytogenetic analysis of mitotic chromosomes has been further defined by synaptonemal complex (SC) analysis and the study of metaphase I chromosome behaviour. This meiotic analysis has allowed a comparison of SC physical lengths, observed chiasma frequencies, and recombination frequencies, as estimated from the genetic map, as a means of comparing physical and genetic distances.

Grazing of the copepod Diaptomus connexus on purple sulphur bacteria in a meromictic salt lake.

A meromictic lake ecosystem (Mahoney Lake, BC, Canada) was investigated to elucidate the significance of chemocline bacteria in the total carbon cycle under natural conditions. In this lake, primary production by oxygenic phototrophs was insufficient to support the observed net secondary production of the calanoid copepod Diaptomus connexus and the rotifer Brachionus plicatilis, indicating the presence of additional food sources for consumers. Mahoney Lake harbours the densest population of phototrophic sulphur bacteria ever reported in a natural body of water. This layer is located at the interface between oxic and anoxic water layers and is dominated by the purple sulphur bacterium Amoebobacter purpureus. The transfer rates of A. purpureus carbon to D. connexus determined in stratified mesocosms were very low (0.71 ngC copepod(-1) day(-1)) and accounted for only 0.6% of the observed net biomass increase in the zooplankter. Stable stratification within the mesocosms prevented an upwelling of A. purpureus into the oxic part. However, measurements of carbon fluxes, infrared fluorescence microscopy and stable carbon analysis provided cumulative evidence that, under in situ conditions, the cell carbon of purple sulphur bacteria indeed enters the aerobic food chain via the grazing activity of D. connexus. Based on a two-source isotopic mixing model, A. purpureus represents at least 75-85% of the diet of D. connexus. Autumnal upwelling into oxic water layers and aggregation of A. purpureus cells appear to be the main factors determining the high carbon flux from purple sulphur bacteria to zooplankton under natural conditions, and most probably also play a key role in other aquatic ecosystems. Through this pathway, over 53% of the reduced organic matter of purple sulphur bacteria trapped in anoxic bottom waters is returned to the oxic realm.

The relationship between genetic and cytogenetic maps of pea. II. Physical maps of linkage mapping populations.

A cytogenetic analysis of inbred lines that have been used to generate genetic maps of pea is presented. Mitotic karyotyping of the inbred lines and meiotic studies of their F1 hybrids have been used to test the prediction that structural differences exist between the parental lines. The results are not compatible with the previously published molecular data. A reordered and updated linkage map of pea is presented that is consistent with the cytogenetic data.