Unusual DNA structures associated with germline genetic activity in Caenorhabditis elegans

We describe a surprising long-range periodicity that underlies a substantial fraction of C. elegans genomic sequence. Extended segments (up to several hundred nucleotides) of the C. elegans genome show a strong bias toward occurrence of AA/TT dinucleotides along one face of the helix while little or no such constraint is evident on the opposite helical face. Segments with this characteristic periodicity are highly overrepresented in intron sequences and are associated with a large fraction of genes with known germline expression in C. elegans. In addition to altering the path and flexibility of DNA in vitro, sequences of this character have been shown by others to constrain DNA::nucleosome interactions, potentially producing a structure that could resist the assembly of highly ordered (phased) nucleosome arrays that have been proposed as a precursor to heterochromatin. We propose a number of ways that the periodic occurrence of An/Tn clusters could reflect evolution and function of genes that express in the germ cell lineage of C. elegans.     

Foraging Habitat Quality Constrains Effectiveness of Artificial Nest-Site Provisioning in Reversing Population Declines in a Colonial Cavity Nester

Among birds, breeding numbers are mainly limited by two resources of major importance: food supply and nest-site availability. Here, we investigated how differences in land-use and nest-site availability affected the foraging behaviour, breeding success and population trends of the colonial cavity-dependent lesser kestrel Falco naumanni inhabiting two protected areas. Both areas were provided with artificial nests to increase nest-site availability. The first area is a pseudo-steppe characterized by traditional extensive cereal cultivation, whereas the second area is a previous agricultural zone now abandoned or replaced by forested areas. In both areas, lesser kestrels selected extensive agricultural habitats, such as fallows and cereal fields, and avoided scrubland and forests. In the second area, tracked birds from one colony travelled significantly farther distances (6.2 km ±1.7 vs. 1.8 km ±0.4 and 1.9 km ±0.6) and had significant larger foraging-ranges (144 km² vs. 18.8 and 14.8 km²) when compared to the birds of two colonies in the extensive agricultural area. Longer foraging trips were reflected in lower chick feeding rates, lower fledging success and reduced chick fitness. Availability and occupation of artificial nests was high in both areas but population followed opposite trends, with a positive increment recorded exclusively in the first area with a large proportion of agricultural areas. Progressive habitat loss around the studied colony in the second area (suitable habitat decreased from 32% in 1990 to only 7% in 2002) is likely the main driver of the recorded population decline and suggests that the effectiveness of bird species conservation based on nest-site provisioning is highly constrained by habitat quality in the surrounding areas. Therefore, the conservation of cavity-dependent species may be enhanced firstly by finding the best areas of remaining habitat and secondly by increasing the carrying capacity of high-quality habitat areas through safe nest-site provisioning.     

Homeostatic control of polyamine levels under long-term salt stress in Arabidopsis: Changes in putrescine content do not alleviate ionic toxicity

Salt stress has been frequently studied in its first osmotic phase. Very often, data regarding the second ionic phase is missing. It has also been suggested that Putrescine or/and Spermine could be responsible for salt resistance. In order to test this hypothesis under long-term salt stress, we obtained Arabidopsis thaliana transgenic plants harboring pRD29A::oatADC or pRD29A::GUS construction. Although Putrescine was the only polyamine significantly increased after salt acclimation in pRD29A::oatADC transgenic lines, this rendered in no advantage to this kind of stress. The higher Spermine levels found in WT and transgenic lines when compared to control conditions along with no increment on Putrescine levels in WT plants under salt acclimation, leads us to analyze Spermine effect on pADC1 and pADC2 expression. Increasing levels of this polyamine inhibits these promoters expression while enhances pRD29A expression, making Spermine the polyamine responsible for salt acclimation, and the transgenic lines developed in this work suitable for studying Putrescine roles in conditions where its biosynthesis would be inhibited in the WT genotype.Key words: arginine decarboxilase, salt acclimation, polyamines, putrescine, spermine, salt overlay sensitive mutants     

Transmission dynamics of heritable silencing induced by double-stranded RNA in Caenorhabditis elegans

Heritable silencing effects are gene suppression phenomena that can persist for generations after induction. In the majority of RNAi experiments conducted in Caenorhabditis elegans, the silencing response results in a hypomorphic phenotype where the effects recede after the F₁ generation. F₂ and subsequent generations revert to the original phenotype. Specific examples of transgenerational RNAi in which effects persist to the F₂ generation and beyond have been described. In this study, we describe a systematic pedigree-based analysis of heritable silencing processes resulting from initiation of interference targeted at the C. elegans oocyte maturation factor oma-1. Heritable silencing of oma-1 is a dose-dependent process where the inheritance of the silencing factor is unequally distributed among the population. Heritability is not constant over generational time, with silenced populations appearing to undergo a bottleneck three to four generations following microinjection of RNA. Transmission of silencing through these generations can be through either maternal or paternal gamete lines and is surprisingly more effective through the male gametic line. Genetic linkage tests reveal that silencing in the early generations is transmitted independently of the original targeted locus, in a manner indicative of a diffusible epigenetic element.     

p38gamma MAPK regulation of glucose transporter expression and glucose uptake in L6 myotubes and mouse skeletal muscle

Skeletal muscle expresses at least three p38 MAPKs (alpha, beta, gamma). However, no studies have examined the potential regulation of glucose uptake by p38gamma, the isoform predominantly expressed in skeletal muscle and highly regulated by exercise. L6 myotubes were transfected with empty vector (pCAGGS), activating MKK6 (MKK6CA), or p38gamma-specific siRNA. MKK6CA-transfected cells had higher rates of basal 2-deoxy-d-[3H]glucose (2-DG) uptake (P < 0.05) but lower rates of 2,4-dinitrophenol (DNP)-stimulated glucose uptake, an uncoupler of oxidative phosphorylation that operates through an insulin-independent mechanism (P < 0.05). These effects were reversed when MKK6CA cells were cotransfected with p38gamma-specific siRNA. To determine whether the p38gamma isoform is involved in the regulation of contraction-stimulated glucose uptake in adult skeletal muscle, the tibialis anterior muscles of mice were injected with pCAGGS or wild-type p38gamma (p38gammaWT) followed by intramuscular electroporation. Basal and contraction-stimulated 2-DG uptake in vivo was determined 14 days later. Overexpression of p38gammaWT resulted in higher basal rates of glucose uptake compared with pCAGGS (P < 0.05). Muscles overexpressing p38gammaWT showed a trend for lower in situ contraction-mediated glucose uptake (P = 0.08) and significantly lower total GLUT4 levels (P < 0.05). These data suggest that p38gamma increases basal glucose uptake and decreases DNP- and contraction-stimulated glucose uptake, partially by affecting levels of glucose transporter expression in skeletal muscle. These findings are consistent with the hypothesis that activation of stress kinases such as p38 are negative regulators of stimulated glucose uptake in peripheral tissues.     

RANK links senescence to stemness in the mammary epithelia,delaying tumor onset but increasing tumor aggressiveness

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Diurnal variation of biological and non-biological particles in the atmosphere of Córdoba, Spain

In this work, the daily distribution of biological and non-biological particles in the atmosphere of the city of Córdoba, Spain, is analysed in order to know at what time of day the concentrations of solid suspended particles in the air is maximum. This knowledge can be useful for all those people who suffer of respiratory diseases and can help them to plan their outdoors activities. The concentrations of non-biological material have been estimated by using spectrophotometrical techniques. With this method, the amount of non-biological material is expressed in percentage of light absorbance in values oscillating from 0.00: absence of particles to 1.00: extreme presence of particles. As the absorption of light depends on the colour of the particle, it is also possible to distinguish dark particles (soot, diesel exhaust particles, sand) from hyaline or light coloured ones (pollen grains and several fungal spores types). The results have shown that the maximum peaks of material are achieved early in the mornings and late in the evenings. Aerobiological methodology has been used when dealing with material of biological origin. In this latter case, pollen grains and fungal spores have been differentiated from the rest of solid material. Due to their importance as agents causing pollen allergy in the area and the high percentage that they represent with respect to the total pollen spectrum, Olea europaea and grass pollen types have been differentiated from the rest of pollen types. The results show that the peaks of non-biological particles in the air throughout the day are related to the activities carried out by men in the city: commercial and working hours or social activities in the different seasons of the year. As regards the biological material, the pattern of fungal spores is clearly influenced by the climatic characteristics of the area, presenting the maximum concentrations in the late afternoon. The pollen grains follow a pattern coinciding with the diurnal maximum temperatures and sunshine hours. The values of the curve oscillate from 200 grains/m3 of air in night hours to maximum over 1000 grains/m3 at midday. The diurnal variation of Olea europaea is quite similar to this general pattern, presenting the maximum concentrations of pollen grains at central hours of day. The grasses show some differences due to the high number of species included in this family.     

Pollen counts statistics and its relevance to precision

In the day to day management of pollen counts from aerobiological samples of national networks, only a small proportion (usually from 12 to 15%) of the daily microscope slide is read. It is generally believed that, otherwise, too much time will be spent reading slides for a minimal increase in precision. Different networks use different slide sampling methods (longitudinal, transverse or at random) and a different number of lines are routinely read. However, the topic of the precision of the different pollen count strategies has seldom been the object of serious investigation. In this study, the precision of different sampling methods of 12 pollen types was investigated by: a) counting pollen grains over the whole slide (3 slides per taxa), b) spatially (i.e. microscope field per microscope field) recording over the 3120 fields found at 400× the location of each pollen grain, c) sub-sampling, by macro procedures, this population by selecting a number of transverse (1 to 48) or longitudinal (1 to 20) lines, or a number of random fields (90 to 2340), so that between 0.96 to 46.15% (transverse), 3 to 66.6% (longitudinal) or 3 to 75% (random) of the whole slide was artificially counted. Between nine and twelve procedures were built per reading strategy. The error found is much higher than what is normally believed, and it was significantly correlated with the abundance of a pollen taxa on the sampled slide. It is only with a total count over 1000 (corresponding to a concentration of above 500 m–3) that the mean error of 4 longitudinal lines (or 13.3% of the slide), the standard protocol of both the Italian Association of Aerobiology (AIA) and the Spanish Aerobiology Network (REA), was always below 30%.