Multi-year evolutionary dynamics of West Nile virus in suburban Chicago,USA, 2005–2007

West Nile virus has evolved in concert with its expansion across North America, but little is known about the evolutionary dynamics of the virus on local scales. We analysed viral nucleotide sequences from mosquitoes collected in 2005, 2006, and 2007 from a known transmission ‘hot spot’ in suburban Chicago, USA. Within this approximately 11 × 14 km area, the viral envelope gene has increased approximately 0.1% yr⁻¹ in nucleotide-level genetic diversity. In each year, viral diversity was higher in ‘residential’ sites characterized by dense housing than in more open ‘urban green space’ sites such as cemeteries and parks. Phylodynamic analyses showed an increase in incidence around 2005, consistent with a higher-than-average peak in mosquito and human infection rates that year. Analyses of times to most recent common ancestor suggest that WNV in 2005 and 2006 may have arisen predominantly from viruses present during 2004 and 2005, respectively, but that WNV in 2007 had an older common ancestor, perhaps indicating a predominantly mixed or exogenous origin. These results show that the population of WNV in suburban Chicago is an admixture of viruses that are both locally derived and introduced from elsewhere, containing evolutionary information aggregated across a breadth of spatial and temporal scales.     

Modeling pattern formation: counting to two in the Drosophila egg

The EGF receptor pathway patterns the Drosophila egg and specifies the position of its dorsal appendages. A new mathematical analysis of this patterning network has highlighted its crucial features and provided novel insights into the spatial and temporal kinetics controlling patterning.     

Disentangling signaling gradients generated by equivalent sources

Yeast cells approach a mating partner by polarizing along a gradient of mating pheromones that are secreted by cells of the opposite mating type. The Bar1 protease is secreted by a-cells and, paradoxically, degrades the α-factor pheromones which are produced by cells of the opposite mating type and trigger mating in a-cells. This degradation may assist in the recovery from pheromone signaling but has also been shown to play a positive role in mating. Previous studies suggested that widely diffusing protease can bias the pheromone gradient towards the closest secreting cell. Here, we show that restricting the Bar1 protease to the secreting cell itself, preventing its wide diffusion, facilitates discrimination between equivalent mating partners. This may be mostly relevant during spore germination, where most mating events occur in nature.     

Disease-driven reduction in human mobility influences human-mosquito contacts and dengue transmission dynamics

Heterogeneous exposure to mosquitoes determines an individual’s contribution to vector-borne pathogen transmission. Particularly for dengue virus (DENV), there is a major difficulty in quantifying human-vector contacts due to the unknown coupled effect of key heterogeneities. To test the hypothesis that the reduction of human out-of-home mobility due to dengue illness will significantly influence population-level dynamics and the structure of DENV transmission chains, we extended an existing modeling framework to include social structure, disease-driven mobility reductions, and heterogeneous transmissibility from different infectious groups. Compared to a baseline model, naïve to human pre-symptomatic infectiousness and disease-driven mobility changes, a model including both parameters predicted an increase of 37% in the probability of a DENV outbreak occurring; a model including mobility change alone predicted a 15.5% increase compared to the baseline model. At the individual level, models including mobility change led to a reduction of the importance of out-of-home onward transmission (R, the fraction of secondary cases predicted to be generated by an individual) by symptomatic individuals (up to -62%) at the expense of an increase in the relevance of their home (up to +40%). An individual’s positive contribution to R could be predicted by a GAM including a non-linear interaction between an individual’s biting suitability and the number of mosquitoes in their home (>10 mosquitoes and 0.6 individual attractiveness significantly increased R). We conclude that the complex fabric of social relationships and differential behavioral response to dengue illness cause the fraction of symptomatic DENV infections to concentrate transmission in specific locations, whereas asymptomatic carriers (including individuals in their pre-symptomatic period) move the virus throughout the landscape. Our findings point to the difficulty of focusing vector control interventions reactively on the home of symptomatic individuals, as this approach will fail to contain virus propagation by visitors to their house and asymptomatic carriers.     

Ammoniated Silver Carbonate for the Demonstration of Lysosomes

Hortega's ammoniated silver carbonate method was used to demonstrate lysosomes in the central nervous system and kidney of adult rats. Formol-CaCl₂, (10%:1%) fixed, frozen sections were impregnated for 10 min in Hortega's solution: 30 ml of 10% AgNO₂ and 90 ml of 5% Na₂CO₃, with concentrated NH₄OH added until the precipitate dissolved, then distilled water to make 400 ml. This procedure revealed silver-positive cytoplasmic structures whose form, shape and distribution were similar to that seen by staining adjacent sections for acid phosphatase. A short fixation of 18-24 hr appears to be essential. A useful, nonenzymatic method for the demonstration of lysosomes is thereby available.     

Polyamines and protein kinase I. Induction of ornithine decarboxylase and activation of protein kinase in rat glioma cells

The activities of cAMP-dependent and independent protein kinases were determined after feeding confluent glioma C6-BU-1 cultures. It has been shown that the activity of both enzymes rose considerably after feeding, and that the ratio of ³²P incorporation into histone, in the absence and the presence of cAMP, was maximal 4 hours after feeding. This increase in protein kinase activity was followed by the activation of ornithine decarboxylase and accumulation of putrescine. Spermine, at millimolar concentrations, inhibited protein kinase, apparently by inactivating the catalytic subunit. It is suggested that this inhibition of protein kinase by polyamines is another regulatory mechanism, which controls cellular growth.     

The effect of starch and of a heteropolysaccharide fraction from Phaseolus vulgaris on development and fecundity of Callosobruchus chinensis (Coleoptera: Bruchidae)

A heteropolysaccharide fraction was isolated from Phaseolus vulgaris beans in which it comprises at least 1% of the dry weight of the beans. This heteropolysaccharide increases larval mortality and decreases the rate of larval development and the number of eggs deposited by females of Callosobruchus chinensis, when incorporated in artificial beans in which the larvae feed. It is composed of galactose, glucose, xylose, arabinose and traces of rhamnose, as determined after acid hydrolysis. Of these individual sugars, arabinose and xylose affect adult fecundity as well. However, partial enzymic hydrolysis of the heteropolysaccharide fraction by C. chinensis larval midgut contents releases only glucose, galactose and trace amounts of arabinose, and the integral structure of heteropolysaccharide may be necessary for biological activity. the incorporation of the starch granules of Phaseolus vulgaris beans into artificial beans increases larval mortality and decreases the rate of larval development of C. chinensis. It is suggested that the heteropolysaccharide fraction as well as the starch are part of a complex of natural components of Phaseolus vulgaris beans that make these beans resistant to C. chinensis.

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Résumé Ce travail cherche à préciser pourquoi les graines de haricot (Phaseolus vulgaris) ne sont pas attaquées par la Buche chinoise (Callosobruchus chinensis), espèce cependant très polyphage.Les graines de haricot renferment 1% d'un hétéropolysaccharide, qui s'est révélé accroître la mortalité et ralentir le développement des larves de cette Bruche, mais qui aussi réduit la fécondité des femelles adultes. Ces observations ont été faites à partir d'élevages sur des milieux artificiels normalement favorables, présentés sous forme de haricots, par un moulage approprié.Une hydrolyse acide de cet hétéropolysaccharide libère du galactose, du glucose, du xylose, de l'arabinose et des traces de rhamnose. Ces sucres simples sont testés: l'arabinose et le rhamnose influencent le développement des larves, tandis que le galactose, le xylose et à nouveau l'arabinose, ont un effet sur la fécondité des femelles (réduction de la fécondité à 40% de la normale avec 1% d'arabinose).Toutefois l'hydrolyse enzymatique de cette fraction hétéropolysaccharidique, par le contenu stomacal de larves de C. chinensis reste partielle et libère seulement du glucose, du galactose et des traces d'arabinose. II est possible que la structure intégrale de l'hétéropolysaccharide soit nécessaire à son activité biologique.Un autre facteur défavorable à C. chinensis pourrait être la nature même des grains d'amidon de Phaseolus. Cet amidon incorporé à l'aliment artificiel accroît en effet la mortalité larvaire et ralentit la vitesse de développement. Cet effet défavorable pourrait être dû à la non digestibilité des grains d'amidon entiers.Il est suggéré que la fraction hétéropolysaccharide et l'amidon de Phaseolus sont deux des facteurs présents dans la graine de haricot qui lui confèrent sa résistance naturelle à la Bruche chinoise.

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Supported in part by the Research and Development Authority of the Hebrew University of Jerusalem.     

CAMKII Activation Is Not Required for Maintenance of Learning-Induced Enhancement of Neuronal Excitability

Pyramidal neurons in the piriform cortex from olfactory-discrimination trained rats show enhanced intrinsic neuronal excitability that lasts for several days after learning. Such enhanced intrinsic excitability is mediated by long-term reduction in the post-burst after-hyperpolarization (AHP) which is generated by repetitive spike firing. AHP reduction is due to decreased conductance of a calcium-dependent potassium current, the sI_AHP. We have previously shown that learning-induced AHP reduction is maintained by persistent protein kinase C (PKC) and extracellular regulated kinase (ERK) activation. However, the molecular machinery underlying this long-lasting modulation of intrinsic excitability is yet to be fully described. Here we examine whether the CaMKII, which is known to be crucial in learning, memory and synaptic plasticity processes, is instrumental for the maintenance of learning-induced AHP reduction. KN93, that selectively blocks CaMKII autophosphorylation at Thr286, reduced the AHP in neurons from trained and control rat to the same extent. Consequently, the differences in AHP amplitude and neuronal adaptation between neurons from trained rats and controls remained. Accordingly, the level of activated CaMKII was similar in pirifrom cortex samples taken form trained and control rats. Our data show that although CaMKII modulates the amplitude of AHP of pyramidal neurons in the piriform cortex, its activation is not required for maintaining learning-induced enhancement of neuronal excitability.