Studies with Differentially Labeled [11C]Cocaine, [11C]Norcocaine, [11C]Benzoylecgonine,and [11C]-and 4′-[18F]Fluorococaine to Probe the Extent to Which [11C]Cocaine Metabolites Contribute to PET Images of the Baboon Brain

Abstract: The psychostimulant drug of abuse, cocaine (benzoylecgonine methyl ester), is rapidly metabolized by cleavage of its two ester groups, to give benzoylecgonine (BE) and ecgonine methyl ester, and by N-demethylation, to give N-norcocaine (NC). The recent use of [N-methyl-¹¹CH₃]cocaine to image brain cocaine binding sites with positron emission tomography (PET) raises the question of whether PET images partially reflect the distribution and kinetics of labeled cocaine metabolites. We prepared [O-metty/-¹¹CH₃]cocaine by methylation of the sodium salt of BE with [¹¹C]CH₃l, and showed that PET baboon brain scans, as well as regional brain kinetics and plasma time-activity curves corrected for the presence of labeled metabolites, are nearly identical to those seen with [N-methyl-¹¹CH₃]cocaine. This strongly suggests that ¹¹C metabolites do not significantly affect PET images, because the metabolite pattern is different for the two labeled forms of cocaine. In particular, nearly half the ¹¹C in blood plasma at 30 min was [¹¹C]CO₂ when [N-methy/-¹¹CH₃]cocaine was administered, whereas [¹¹C]CO₂ was not formed from [O-methy/-¹¹CH₃]cocaine. Only a trace of [¹¹C]NC was detected in plasma after [O-methyl-¹¹CH₃]cocaine administration. Nearly identical brain PET data were also obtained when 4′-[N-methy/-¹¹CH₃]fluorococaine and 4′-[¹⁸F]fluoro-cocaine (prepared by nucleophilic aromatic substitution from [¹⁸F]fluoride-and 4′-nitrococaine) were compared with [N-methy/-¹¹CH₃]cocaine. In vitro assays with rat brain membranes showed that cocaine and 4′-fluoroco-caine were equipotent at the dopamine reuptake site, but that 4′-fluorococaine was about 100 times more potent at the 5-hydroxytryptamine reuptake site. The studies with positron-emitting 4′-fluorococaines thus support the lack of significance of labeled metabolites or of binding to 5-hydroxytryptamine reuptake sites to PET images taken with [N-methy/-¹¹CH₃]cocaine. [¹¹C]NC prepared by O-methylation of norbenzoylecgonine gave PET images with preferential uptake in striatum, but slower clearance from all brain regions than [O-methy/-¹¹CH₃]cocaine. [¹¹C]BE prepared by N-methylation of norbenzoylecgonine did not show brain uptake.     

Identifying plant functional types using floristic data bases: Ecological correlates of plant range size

Abstract. In an effort to identify ‘plant functional types’, the islands floras of Great Britain and Kríti (Crete, Greece) were examined separately for ecological correlates of plant range size. Plant functional types (PFTs) were defined here as categories into which plants could be grouped on the basis of attributes that predict greater or lesser sensitivity to ecological variability. Plant range size indicates commonness of a species and was assumed to be a proxy for ‘ecological flexibility’, i.e. species of larger range sizes can better withstand environmental change and differences than species of smaller range sizes. Using evolutionary comparative methods that account for the effect of taxonomic relatedness, both floras were investigated for the effects on range size of woodiness vs. non-woodiness, trees vs. shrubs, trees vs. herbs and shrubs vs. herbs. The British flora was examined additionally for the effects of wind- vs. non-wind-pollination, self vs. animal pollination and animal vs. non-animal fruit dispersal on range size. Two analyses showed significant effects on range size: for British species, trees had larger ranges than shrubs, and wind- pollinated species had larger ranges than non-wind-pollinated species. It is suggested that the lack of a similar pattern for shrubs and trees in Kríti is because the lower water availability of Kríti imbues shrubs with an ecophysiological advantage not relevant in plants of Great Britain. That trees have larger range sizes than shrubs in Great Britain is ascribed to the greater importance of competition for light when other factors are not at issue. The greater range of wind-pollinated than non-windpollinated species in Great Britain is postulated to be because both mutualists must be capable of invading new areas. This may be termed a ‘cost of mutualism’. In terms of PFTs, the results indicate that ‘life-form’ is too broad a classification category by which to differentiate relative sensitivity to environmental variability in Great Britain, in that there were significant differences in range size of trees and shrubs, but not between either of the two categories and herbs, or between woody and non-woody plants. Although pollination type may predict relative sensitivity to variation in Great Britain, dispersal type will not. Finally, differences between Great Britain and Kríti in relative range size patterns suggests that plant functional types may be specific to a region or set of conditions.     

Three patterns of mitochondrial DNA nucleotide divergence in the meadow vole,Microtus pennsylvanicus

Summary The DNA sequence was determined for the cytochrome c oxidase II (COII), tRNA^Lys, and ATPase 8 genes from the mitochondrial genome of the meadow vole, Microtus pennsylvanicus. When compared to other rodents, three different patterns of evolutionary divergence were found. Nucleotide variation in tRNA^Lys is concentrated in the TC loop. Nucleotide variation in the COII gene in three genera of rodents (Microtus, Mus, Rattus) consists predominantly of transitions in the third base positions of codons. The predicted amino acid sequence in highly conserved (>92% similarity). Analysis of the ATPase 8 gene among four genera (Microtus, Cricetulus, Mus, Rattus) revealed more detectable transversions than transitions, many fixed first and second position mutations, and considerable amino acid divergence. The rate of nucleotide substitution at nonsynonymous sites in the ATPase 8 gene is 10 times the rate in the COII gene. In contrast, the estimated absolute mutation rate as determined by analysis of nucleotide substitutions at fourfold degenerate sites probably is the same for the two genes. The primary sequences of the ATPase 8 and COII peptides are constrained differently, but each peptide is conserved in terms of predicted secondary-level configuration.     

Phenotypic profiling of mGlu7 knockout mice reveals new implications for neurodevelopmental disorders

Neurodevelopmental disorders are characterized by deficits in communication, cognition, attention, social behavior and/or motor control. Previous studies have pointed to the involvement of genes that regulate synaptic structure and function in the pathogenesis of these disorders. One such gene, GRM7, encodes the metabotropic glutamate receptor 7 (mGlu₇), a G protein‐coupled receptor that regulates presynaptic neurotransmitter release. Mutations and polymorphisms in GRM7 have been associated with neurodevelopmental disorders in clinical populations; however, limited preclinical studies have evaluated mGlu₇ in the context of this specific disease class. Here, we show that the absence of mGlu₇ in mice is sufficient to alter phenotypes within the domains of social behavior, associative learning, motor function, epilepsy and sleep. Moreover, Grm7 knockout mice exhibit an attenuated response to amphetamine. These findings provide rationale for further investigation of mGlu₇ as a potential therapeutic target for neurodevelopmental disorders such as idiopathic autism, attention deficit hyperactivity disorder and Rett syndrome.     

Addition of Nitrogen Increases Variability of Vegetation Cover in an Arid System with Unpredictable Rainfall

Ecosystems - Addition of nitrogen (N) to rangeland that has been degraded through overgrazing or drought can hasten vegetation recovery. Additional N may influence temporal stability of vegetation...     

Linking mosquito surveillance to dengue fever through Bayesian mechanistic modeling

Our ability to effectively prevent the transmission of the dengue virus through targeted control of its vector, Aedes aegypti, depends critically on our understanding of the link between mosquito abundance and human disease risk. Mosquito and clinical surveillance data are widely collected, but linking them requires a modeling framework that accounts for the complex non-linear mechanisms involved in transmission. Most critical are the bottleneck in transmission imposed by mosquito lifespan relative to the virus’ extrinsic incubation period, and the dynamics of human immunity. We developed a differential equation model of dengue transmission and embedded it in a Bayesian hierarchical framework that allowed us to estimate latent time series of mosquito demographic rates from mosquito trap counts and dengue case reports from the city of Vitória, Brazil. We used the fitted model to explore how the timing of a pulse of adult mosquito control influences its effect on the human disease burden in the following year. We found that control was generally more effective when implemented in periods of relatively low mosquito mortality (when mosquito abundance was also generally low). In particular, control implemented in early September (week 34 of the year) produced the largest reduction in predicted human case reports over the following year. This highlights the potential long-term utility of broad, off-peak-season mosquito control in addition to existing, locally targeted within-season efforts. Further, uncertainty in the effectiveness of control interventions was driven largely by posterior variation in the average mosquito mortality rate (closely tied to total mosquito abundance) with lower mosquito mortality generating systems more vulnerable to control. Broadly, these correlations suggest that mosquito control is most effective in situations in which transmission is already limited by mosquito abundance.