Detection of Plasmodium falciparum DNA in a microtitration plate-based hybridization test

A rapid DNA-test, depending on the affinity based hybrid collection principle, was developed for the detection of Plasmodium falciparum DNA from clinical specimens. In this method, hybridization takes place in solution and the hybrids are collected onto a solid phase for measurement. Two probes are used, one labelled with an affinity tag (biotin) and the other with a detectable label (32P). In the present test a single oligonucleotide complementary to a 21-base pair sequence which is highly repeated in the parasite genome served both as capture and detector probe. The test is a 2-h hybridization performed in streptavidin coated microtitration plate wells, onto which the labelled hybrids simultaneously bind. The sensitivity of the assay with a crude erythrocyte lysate specimen was 1.6 x 10(9) repeat units corresponding to about 160 parasites in one microliter blood. The results allowed quantification of the repeat sequences and thus estimation of the degree of parasitemia in clinical specimens.     

Accumulation of pyrophosphate correlates with the increased level of nucleoside triphosphates in Escherichia coli

We measured the concentration of nucleoside triphosphates and inorganic pyrophosphate in Escherichia coli in conditions where nucleotide synthesis or nucleic acid synthesis was inhibited. The inhibitors that brought about an accumulation of some of the four ribonucleoside triphosphates also increased the pyrophosphate level. In a pyrimidine auxotrophic strain uracil starvation led to simultaneous accumulation of ATP and pyrophosphate, and they both rapidly returned to normal level when starvation was relieved. These results indicate the possible involvement of pyrophosphate in the reactions leading to the accumulation of nucleoside triphosphates.     

Phylogenetic analysis of cuckoo wasps (Hymenoptera: Chrysididae) reveals a partially artificial classification at the genus level and a species‐rich clade of bee parasitoids

Cuckoo wasps (Hymenoptera: Chrysididae) are a species‐rich family of obligate brood parasites (i.e. parasitoids and kleptoparasites) whose hosts range from sawflies, wasps and bees, to walking sticks and moths. Their brood parasitic lifestyle has led to the evolution of fascinating adaptations, including chemical mimicry of host odours by some species. Long‐term nomenclatural stability of the higher taxonomic units (e.g. genera, tribes, and subfamilies) in this family and a thorough understanding of the family's evolutionary history critically depend on a robust phylogeny of cuckoo wasps. Here we present the results from phylogenetically analysing ten nuclear‐encoded genes and one mitochondrial gene, all protein‐coding, in a total of 186 different species of cuckoo wasps representing most major cuckoo wasp lineages. The compiled data matrix comprised 4946 coding nucleotide sites and was phylogenetically analysed using classical maximum‐likelihood and Bayesian inference methods. The results of our phylogenetic analyses are mostly consistent with earlier ideas on the phylogenetic relationships of the cuckoo wasps' subfamilies and tribes, but cast doubts on the hitherto hypothesized phylogenetic position of the subfamily Amiseginae. However, the molecular data are not fully conclusive in this respect due to low branch support values at deep nodes. In contrast, our phylogenetic estimates clearly indicate that the current systematics of cuckoo wasps at the genus level is artificial. Several of the currently recognized genera are para‐ or polyphyletic (e.g. Cephaloparnops, Chrysis, Chrysura, Euchroeus, Hedychridium, Praestochrysis, Pseudochrysis, Spintharina, and Spinolia). At the same time, our data support the validity of the genus Colpopyga, previously synonymized with Hedychridium. We discuss possible solutions for how to resolve the current shortcomings in the systematics of cuckoo wasp genera and decided to grant Prospinolia the status of a valid genus (Prospinolia stat.n. ) and transferring Spinolia theresae [du Buysson 1900] from Spinolia to Prospinolia (Prospinolia theresae stat.restit. ). We discuss the implications of our phylogenetic inferences for understanding the evolution of host associations in this group. The results of our study not only shed new light on the evolutionary history of cuckoo wasps, but also set the basis for future phylogenomic investigations on this captivating group of wasps by guiding taxonomic sampling efforts and the design of probes for target DNA enrichment approaches.     

A method to estimate body mass and relative age of exotic lagomorphs in the southern Neotropics

Exotic prey species can dramatically alter trophic interactions by functionally replacing native prey species. This pattern has been observed in the southern Neotropics, where introduced haresLepus europaeus Pallas, 1778 and rabbitsOryctolagus cuniculus (Linnaeus, 1758) have become the staple prey for native predators. Despite their importance as prey, no data are available on the mass and relative age of the lagomorphs on which native predators feed. We used linear regression models to predict the mass of lagomorphs from their hind-foot length, and investigated the influence of sex, season, and collection site on this relationship. Hind-foot length was a good predictor of body mass for both species and accounted for 58.4 and 71.6% of the variability in body mass for hares and rabbits, respectively. Sex and season significantly influenced the relationship, whereas the effect of collection site was negligible. Hind-foot length was also a good predictor of age class for hares and rabbits, allowing the discrimination between nonreproductive and reproductive classes for both species. Future research can use this method to determine the biomass and age class of exotic lagomorphs consumed by predators.     

Winter Conditions and Land Cover Structure the Subnivium,A Seasonal Refuge beneath the Snow

In seasonally snow-covered environments, many organisms endure winter by using the subnivium, a below-snow thermally stable seasonal refugium. Because the insulation of snow is dependent on snow depth and density, the stability of temperatures within the subnivium varies across land cover types. Additionally, across much of the Northern Hemisphere snow extent, depth and duration are generally decreasing while snow density is increasing due to climate change. These changes are likely to destabilize the thermal profile of the subnivium, although they have not yet been quantified. To explore the effects of land cover and climate change on the subnivium, we measured snow pack characteristics (depth and density), and ambient and subnivium temperatures from three different land cover types (prairie, deciduous forest, and coniferous forest) and within a micro-greenhouse (2.5 x 2.5 x 2 m) that maintained a temperature of 5°C warmer than outdoor ambient temperatures, and automatically opened during snow events throughout the winter of 2013/14. We found that the mean daily subnivium temperature was significantly colder in the deciduous cover type than the prairie cover type, and that prairie had higher maximum subnivium temperatures than both of the other cover types. Our climate change simulation revealed that, although ambient temperatures within the micro-greenhouse were 5°C warmer than outside the greenhouse, the daily minimum subnivium temperature was significantly lower inside the greenhouse. Our findings suggest that climate change could have considerable effects on the refuge quality of the subnivium, and that some cover types appear to be more susceptible to these effects than others.     

Effect of denucleation and UV-irradiation on the subcellular morphology inMicrasterias

Summary The effects of the elimination of the nuclear control on the ultrastructure of the green algaMicrasterias torreyi. Bail, have been studied by using centrifugation for denucleation and lethal dose of UV-light. Centrifugated anucleate cells were fixed 7 and 26 hours after the treatment and the UV-treated cells 4 and 8 hours after the irradiation. Although both treatments eliminate the nuclear control and the treated cells resemble morphologically each other, yet there are differences in ultrastructure suggesting that they are also brought about by other factors than the presence of nucleus. Both the treatments cause accumulation of cell wall material in the tips of lobes. The cell wall shows unusual secondary thickening with electron dense spots embedded in the matrix. The denucleation retards the functional cycle of Golgi apparatus and the production of vesicles has stopped in the 26-hour-denucleated cells. It is possible that flat vesicle production is totally absent in denucleated cells.First the UV-treatment seems to activate the function of Golgi apparatus but later on the vesicle production almost stops. It seems to eliminate the production of large vesicles but not that of dark vesicles.Both the treatments cause deterioration of ER membranes and polysomes, and in consequence, probably inhibit protein synthesis.Unlike UV-irradiation, denucleation appears to destroy the microtubule system. Mitochondrial cristae have almost entirely vanished within 26 hours after denucleation. Effect of denucleation and UV-irradiation on the subcellular morphology inMicrasterias.     

Responses to the lowering of magnesium and calcium concentrations in the cerebrospinal fluid of unanesthetized sheep.

A technique for ventriculolumbar perfusion of the cerebrospinal fluid space has been used to study the neuromuscular effects of low concentrations of magnesium and calcium in the cerebrospinal fluid of conscious sheep. Perfusion with synthetic cerebrospinal fluid solutions containing less than 0-6 mg magnesium/100 ml produced episodes of tetany which were abolished by perfusion with a solution of normal magnesium concentration. This suggests that the low cerebrospinal fluid magnesium concentrations reported in cases of hypomagneseamic tetany may result in changes within the central nervous system that could produce the nervous signs. Perfusates with a calcium concentration below 2-0 mg/100 ml caused hyperpnoea and continuous muscle tremors. Magnesium (0-6 mg/100 ml) and calcium (2-0 mg/100 ml) perfused simultaneously acted synergistically to produce signs characteristic of low levels of each of the ions.