Identification of new human origins of DNA replication by an origin-trapping assay

Metazoan genomes contain thousands of replication origins, but only a limited number have been characterized so far. We developed a two-step origin-trapping assay in which human chromatin fragments associated with origin recognition complex (ORC) in vivo were first enriched by chromatin immunoprecipitation. In a second step, these fragments were screened for transient replication competence in a plasmid-based assay utilizing the Epstein-Barr virus latent origin oriP. oriP contains two elements, an origin (dyad symmetry element [DS]) and the family of repeats, that when associated with the viral protein EBNA1 facilitate extrachromosomal stability. Insertion of the ORC-binding human DNA fragments in oriP plasmids in place of DS enabled us to screen functionally for their abilities to restore replication. Using the origin-trapping assay, we isolated and characterized five previously unknown human origins. The assay was validated with nascent strand abundance assays that confirm these origins as active initiation sites in their native chromosomal contexts. Furthermore, ORC and MCM2-7 components localized at these origins during G(1) phase of the cell cycle but were not detected during mitosis. This finding extends the current understanding of origin-ORC dynamics by suggesting that replication origins must be reestablished during the early stages of each cell division cycle and that ORC itself participates in this process.     

Large-scale Metabolomic Profiling Identifies Novel Biomarkers for Incident Coronary Heart Disease

Analyses of circulating metabolites in large prospective epidemiological studies could lead to improved prediction and better biological understanding of coronary heart disease (CHD). We performed a mass spectrometry-based non-targeted metabolomics study for association with incident CHD events in 1,028 individuals (131 events; 10 y. median follow-up) with validation in 1,670 individuals (282 events; 3.9 y. median follow-up). Four metabolites were replicated and independent of main cardiovascular risk factors [lysophosphatidylcholine 18∶1 (hazard ratio [HR] per standard deviation [SD] increment = 0.77, P-value<0.001), lysophosphatidylcholine 18∶2 (HR = 0.81, P-value<0.001), monoglyceride 18∶2 (MG 18∶2; HR = 1.18, P-value = 0.011) and sphingomyelin 28∶1 (HR = 0.85, P-value = 0.015)]. Together they contributed to moderate improvements in discrimination and re-classification in addition to traditional risk factors (C-statistic: 0.76 vs. 0.75; NRI: 9.2%). MG 18∶2 was associated with CHD independently of triglycerides. Lysophosphatidylcholines were negatively associated with body mass index, C-reactive protein and with less evidence of subclinical cardiovascular disease in additional 970 participants; a reverse pattern was observed for MG 18∶2. MG 18∶2 showed an enrichment (P-value = 0.002) of significant associations with CHD-associated SNPs (P-value = 1.2×10⁻⁷ for association with rs964184 in the ZNF259/APOA5 region) and a weak, but positive causal effect (odds ratio = 1.05 per SD increment in MG 18∶2, P-value = 0.05) on CHD, as suggested by Mendelian randomization analysis. In conclusion, we identified four lipid-related metabolites with evidence for clinical utility, as well as a causal role in CHD development.     

The Time Distribution of Sulfadoxine-Pyrimethamine Protection from Malaria

Sulfadoxine-pyrimethamine (SP) has been one of the most widely used antimalarial treatments world-wide, and is also used prophylactically in vulnerable populations. In this paper, we develop a mathematical model which allows us to infer the time distribution of SP protection from drug-trial data. Fitting our model to data from a controlled field study in Mali, we find that SP provided protection from malaria for an average of 37.9?days in this pediatric population. We demonstrate that the duration of SP protection is not well described by an exponential distribution, and in fact has a much narrower dispersal about the mean; the best-fit standard deviation predicted by our model was only 17.0?days, as opposed to 41.8?days for the exponential model. We estimate the monthly entomological inoculation rate and the basic reproductive number for malaria in this population, and demonstrate that extremely high SP treatment rates would be necessary to maintain an effective reproductive number below one throughout a single rainy season. These results have implications for further efforts to model the impact of SP treatment, or for investigations of the optimal timing of prophylactic SP.     

Effect of prolonged root hypoxia on the antioxidant content of tomato fruit

Tomato fruit quality depends on its metabolite content, which in turn is determined by numerous metabolic changes occurring during fruit development and ripening. The aim of this work was to investigate whether flooding affects the nutritional quality of tomato fruit, focusing on compounds essential to human health: carotenoids and ascorbate. To this end, tomato plants (Solanum lycopersicum L. cv Micro-Tom) were submitted to prolonged root hypoxia (1–2% O₂) at first flower anthesis. Fruits were harvested at five stages of the ripening process and analysed for their carotenoid and ascorbate contents. Our results showed that the ripening of fruits that developed on hypoxia treated plants was not inhibited. However, root hypoxia significantly limits carotenoid and ascorbate accumulation in pericarp during fruit ripening, the strongest effects being observed at late stages of ripening. Limitation of both carotenoids and ascorbate accumulation seems to be primarily mediated by the reduced level of expression of genes of the corresponding metabolic pathway.     

Effets in vitro des dermaseptines sur les caractéristiques du sperme

The spermicidal efficacy of synthetic peptides, dermaseptin (DS1) and (DS4), was studied under in vitro conditions using human spermatozoa. The data showed that sperm motility was inhibited with various concentrations of dermaseptins at different intervals ranging from 2 to 240 min. The effective 100% inhibitory concentration (EC100) of DS4 sperm immobilization assay was equal to 50 mg/ml at 30 min, while the EC 100 of DS1 was equal to 100 mg/ml. The presence of 0.1% of chelating agent, EDTA, reduced the EC100 of DS4 to 5 mg/ml, while less than a twofold enhancement in DS 1 activity was observed in combination with EDTA. The action of dermaseptins on sperm motility was observed to be dose-dependent. Addition of pentoxifylline, which is known to enhance sperm motility, and Ca²⁺, which is a key element for sperm movement, did not prevent the spermicidal action of dermaseptins.     

Old and new host-parasitoid associations: parasitism of the invasive fruit fly Bactrocera invadens (Diptera: Tephritidae) and five African fruit fly species by Fopius arisanus,an Asian opiine parasitoid

Fopius arisanus (Sonan), a solitary koinobiont endoparasitoid of fruit flies, was introduced for testing and final release against the recently discovered species Bactrocera invadens Drew, Tsuruta and White in Africa. Laboratory experiments were conducted to assess host preference, host acceptability for oviposition, and physiological suitability of B. invadens and five other indigenous tephritid fruit fly species, namely, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), C. cosyra (Walker), C. rosa Karsch, C. fasciventris (Bezzi), and C. anonae Graham. Females of F. arisanus visited all host egg patches, but showed a stronger preference to eggs of B. invadens, which was also most accepted for oviposition. Successful development of parasitoid progenies varied greatly across hosts, with B. invadens yielding the highest parasitoid progeny and C. fasciventris yielding no F. arisanus progeny. Most of the parasitoid eggs laid in C. rosa and C. fasciventris were encapsulated. Sex ratio was not influenced by host species and it was female biased in all hosts that produced parasitoid progeny. Fopius arisanus was able to establish a new association with C. capitata, C. cosyra and to a lesser extent C. anonae. The results are discussed in the light of the potential use of F. arisanus as a biological control agent of B. invadens.     

Vasopressin analog (DDAVP) facilitates concept learning in human males

The effects of desmopressin acetate (DDAVP) were studied in normal males. Subjects were given 60μg of DDAVP, a placebo, or no treatment and were given several behavioral tests. DDAVP enhanced learning of all problems on a concept shift task but had no effect on visual memory, anxiety, blood pressure or heart rate. It was suggested that DDAVP may influence memory via its actions on attention.     

Laboratory studies on the biology of the corn leaf aphid, Rhopalosiphum maidis (Homoptera: Aphididae)

The biology of R. maidis has been shown to be affected considerably by the temperature, the host plant, and the physiological age of food plant. The optimum temperature was 30°C; here nymphal development was accelerated, the whole life-span was drastically shortened to less than half that at 15°, and natality rate/surviving female was increased with rather negligible deaths among progeny. Barley was the most favourable host studied under the specified experimental conditions, and young plants (3–5 days old) represented the best stage of plant growth. R. maidis may have about 50 generations/year when aphids were kept feeding continuously on 5-day old barley plants but this does, by no means, represent the actual number of generations under field conditions; it only indicates that almost half of these may be restricted to the summertime.

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Résumé La présente étude montre que la température, ainsi que l'espèce de la plante hôte et son âge physiologique influencent grandement la biologie de Rhopalosiphum maïdis.Quand ce puceron est élevé sur des pousses d'orge âgées de 5 jours, la température optimum est de 30°. Dans ces conditions, la maturation peut être acquise en un délai aussi court que 4,6 jours, tandis que la longévité est réduite à 16,8±1,3 jours contre 35,1±5,1 jours à 15°. Enfin, la natalité par femelle et par jour est accrue avec un taux de mortalité négligeable parmi la descendance.De quatre plantes testées, (orge, mil, fève et maïs) au même âge de croissance (7 jours) l'orge est le plus favorable. Rhopalosiphum maïdis peut avoir environ 50 générations par an quand il est élevé de façon continue sur des plants d'orge âgés de 5 jours; cela ne signifie pas que dans les conditions naturelles il en est ainsi, mais indique que presque la moitié de ce nombre de générations peut intervenir pendant l'été.

     

Characterization of Lead in Soils of a Rifle/Pistol Shooting Range in Central Florida,USA

The distribution of lead in soil samples collected from both surface (0 to 10?cm) and profile (O 0 to 10?cm, E 11 to 30?cm, Eb 31 to 50?cm, Bw 51 to 100?cm, and C 181 to 200?cm) at a 14-year-old rifle/pistol shooting range located in central Florida were determined using EPA Method 3051a (microwave, HNO₃/HCl=3:1, v/v). In addition to total lead analysis, Toxicity Characteristic Leaching Procedure (TCLP) analysis was performed on corresponding samples to determine whether the soils would require special handling as hazardous waste if the soils were to be removed from the range. Total lead in surface soils varied from 330 to 17 850?mg Pb kg^?1, with the greatest concentration in the middle of the backstop berm. The TCLP tests indicated that lead in all surface soils exceeded the 5?mg Pb L^?1 critical level of federal regulation for solid wastes and hazardous wastes provided by the Resource Conservation and Recovery Act (RCRA) and would be characterized as hazardous waste. Sequential fractionation and X-ray diffraction (XRD) analyses revealed that lead carbonate existed predominantly (91.3%) in the berm soil. The weathering of lead bullets in the soil environments formed primarily as hydrocerussite (Pb₃(CO₃)₂(OH)₂), with small amounts of massicot (PbO) and cerussite (PbCO₃). However, the elevated soil pH, caused by the oxidization and transformation process of elemental lead in lead bullets, could be a significant factor in limiting the migration of lead in the soil.