Two novel prevalent polymorphisms in the hormone-sensitive lipase gene have no effect on insulin sensitivity of lipolysis and glucose disposal.

Free fatty acids released during triglyceride lipolysis play an important role in obesity-associated insulin resistance of glucose disposal. Individual sensitivity of lipolysis to the suppressive effect of insulin varies greatly among healthy subjects. It is possible that genetic factors contribute to this variation. Among the many proteins involved in the regulation of lipolysis, hormone-sensitive lipase (HSL) represents a prime candidate for genetic variants contributing to the biological variation of insulin sensitivity of lipolysis. We determined the insulin sensitivity of lipolysis (suppression of isotopically [primed-continuous infusion of d5 glycerol] measured glycerol rate of appearance) and of glucose disposal, using a three-step (n = 20) or standard (n = 53) hyperinsulinemic euglycemic clamp in 73 healthy, unrelated subjects. To assess the possible role of genetic polymorphisms, we directly sequenced the coding region of the HSL gene and the noncoding exon B from these subjects. We identified two silent mutations and three amino acid polymorphisms: Arg262Met (prevalence, 5%), Glu620Asp (prevalence, 31%) and Ser681Ile (prevalence, 22%). The latter two are located in the regulatory domain of HSL but neither had a significant impact on insulin sensitivity of lipolysis or glucose disposal (with and without adjustment for obesity and age as covariates; all P values > 0.20). We conclude that a number of genetic polymorphisms in HSL exist, some of which are highly prevalent. Neither of the polymorphisms we identified in the coding region, however, contributed measurably to the biological variation of insulin sensitivity in our lean, healthy population.     

Uncertain biotic and abiotic interactions in benthic communities

We analyze marine benthic communities at different sites in Skagerrak with the purpose of understanding the role of exogenous and endogenous factors in explaining the species' temporal dynamics. The previous finding that the dynamics of these species communities are mainly driven and synchronized by environmental (temperature) forcing was only weakly supported when analyzing single-species dynamics at five sites where four of the species were present every year. There was no consistent pattern in how the temperature affected the realized per capita growth rate, either across species at a given site, or among sites for a given species. Furthermore, there was no net-interaction from the community on a given species strong enough to give rise to second-order dynamics. However, when implementing a Multi Dimensional Scaling (MDS) analysis and incorporating all sampling sites and species -we found that the different communities clustered in relation to depth, hence, communities at the same depth were more "similar" than communities at different depth. Revealing the underlying interactions shaping these marine benthic communities is a challenge that calls for an array of various and complementary approaches.     

Ultrastructural studies of dorsal ciliated organs in Spionidae (Annelida: Polychaeta)

The distribution and structural components of dorsal ciliated organs (dco) in 15 species of the Spionidae were studied by scanning- and transmission electron microscopy. Based on the distribution patterns of dco, the investigated species are divided into four non-systematic groups: (I) paired anterior dco, (II) paired dco extending posteriorly for several chaetigers, (III) paired anterior dco in combination with unpaired, sexually dimorphic, metameric dco, and (IV) paired anterior dco in combination with paired, metameric dco. Previous ultrastructural studies have only included species possessing organs of groups I and III. In the present investigation the ultrastructure of dco found in Laonice bahusiensis and Spio cf. filicornis (species with dco of groups II and IV) is studied in an attempt to consider their homology. Apart from the metameric dco of group III, similarities of the cellular components of the dco indicate a homology to nuchal organs.     

1-substituted cyclopropenes: Effective Blocking Agents for Ethylene Action in Plants

A series of 1-alkane substituted cyclopropenes has been prepared and tested as ethylene antagonists using banana fruits as an assay system. 1-Methyl-, 1-ethyl-, 1-propyl-, 1-butyl-, 1-pentyl-, 1-hexyl-, 1-heptyl-, 1-octyl-, 1-nonyl-, and 1-decylcyclopropene were all very active compounds. 1-Methylcyclopropene protected bananas from ethylene with a minimum concentration of 0.7 nl.l^–1 after a 24 h exposure. As the carbon chain length was extended the minimum requirement increased some, but starting with 1-butylcyclopropene, the minimum concentration requirement declined and many cyclopropenes were required in lower concentrations than 1-methylcyclopropene. The time of protection at ambient temperature (22–23 °C) was 12 d for 1-methyl-, 1-ethyl-, 1-propyl-, and 1-butylcyclopropene. 1-Pentylcyclopropene protected bananas for 14 d, 1-hexylcyclopropene for 20 d, 1-heptylcyclopropene for 21 d, 1-octylcyclopropene for 25 d, 1-nonylcyclopropene for 35 d, and 1-decylcyclopropene for 36 d.     

Local enrichment with homopolymeric (dA/dT) DNA in genomes of some lower dipterans and Drosophila melanogaster

An investigation into the chromosomal localization of homopolymeric dA/dT was carried out with species of the genera Rhynchosciara, Chironomus, Drosophila and several other taxa. In situ hybridisation probing mitotic and polytene chromosomes with RNA homopolymers was performed, followed by immunological detection of the DNA/RNA hybrid. Use of this method allowed us to assess specific regions of some dipteran genomes, where the signal was generally, but not always, located in heterochromatic regions. Human and Drosophila chromosome regions known to contain dA/dT runs of up to 153 bp were devoid of consistent labelling. The stability of the rA/dT hybrid formed in situ was in agreement with the T(m) for long rA/dT hybrid complexes, suggesting that the method used in this work is able to identify unusually long homopolymeric dA/dT tracts.     

Effects of Barriers and Thermal Refugia on Local Movement of the Threatened Leopard Darter, Percina pantherina

Local, short-term dispersal by the U.S. federally-threatened leopard darter, Percina pantherina, was examined in the field and in the laboratory to assess the possible effects of natural versus man-made barriers on movement. Mark-resight studies were conducted in two summers at sites in the Glover River (southeastern Oklahoma, U.S.A.). At one site, patches of 'preferred' habitat were separated by a natural riffle; at the other site, by a low-water road crossing with culverts. At the Natural Riffle site, darters moved downstream across the riffle, but also moved upstream into deeper water when water temperatures exceeded 29°C in the `preferred' habitat. Use of deeper, cooler waters by this species in late summer suggests that thermal refugia may be important habitats for the long-term management of leopard darters. At the Road Crossing site, all documented movement was in a downstream direction, and at least two darters traversed culverts in the low-water bridge. Laboratory studies of movement across several types of culverts suggested that culverts significantly decrease the probability of movement among habitat patches.     

A Plasmid Selection System in Lactococcus lactis and Its Use for Gene Expression in L. lactis and Human Kidney Fibroblasts

We report the development of a nonantibiotic and nonpathogenic host-plasmid selection system based on lactococcal genes and threonine complementation. We constructed an auxotrophic Lactococcus lactis MG1363Δthr strain which carries deletions in two genes encoding threonine biosynthetic enzymes. To achieve plasmid-borne complementation, we then constructed the minimal cloning vector, pJAG5, based on the genes encoding homoserine dehydrogenase-homoserine kinase (the hom-thrB operon) as a selective marker. Using strain MG1363Δthr, selection and maintenance of cells carrying pJAG5 were obtained in threonine-free defined media. Compared to the commonly used selection system based on erythromycin resistance, the designed complementation system offers a competitive and stable plasmid selection system for the production of heterologous proteins in L. lactis. The potential of pJAG5 to deliver genes for expression in eukaryotes was evaluated by insertion of a mammalian expression unit encoding a modified green fluorescent protein. The successful delivery and expression of genes in human kidney fibroblasts indicated the potential of the designed nonantibiotic host-plasmid system for use in genetic immunization.     

Increased frequency of scale anomalies and loss of genetic variation in serially bottlenecked populations of the dice snake, Natrix tessellata

Documented demographic bottlenecks resultingfrom introductions of the dice snake to severallakes in Switzerland provide a rare opportunityto study the effect of serial bottlenecks onthe genetic properties of Natrixtessellata populations. We investigated twointroduced populations using informationderived from eight microsatellite markers. Bothintroduced populations had significantlyreduced levels of allelic diversity relative tonon-bottlenecked populations. The severity ofthe bottlenecks was underlined by thesignificant reduction in observed and expectedheterozygosity. The loss of allelic diversityand observed heterozygosity was stronger in theserially bottlenecked population than in thepopulation that was bottlenecked only once.From previous studies, scale anomalies wereknown to be more common in introducedpopulations relative to native populations. Weinvestigated whether the higher occurrence ofscale anomalies in introduced populations isassociated with individual heterozygosity andmean genomic diversity d ². We founda significant relationship between theoccurrence of scale anomalies and individualheterozygosity but no significant relationshipbetween scale anomalies and the microsatellitespecific measurement, d ², was found.Because of their known history, introducedpopulations in Switzerland may serve as a modelto demonstrate the effect of severe populationbottlenecks on genetic variability anddevelopmental stability in N. tessellata.The results therefore help to device strategiesfor the management and protection of endangerednatural N. tessellata populations.     

A theoretical approach to predicting the success of genetic manipulation of malaria mosquitoes in malaria control

Background  

Mosquitoes that have been genetically modified to better encapsulate the malaria parasite Plasmodium falciparum are being considered as a possible tool in the control of malaria. Hopes for this have been raised with the identification of genes involved in the encapsulation response and with advances in the tools required to transform mosquitoes. However, we have only very little understanding of the conditions that would allow such genes to spread in natural populations.