Synthesis and biological evaluation of novel propargyl amines as potential fluorine-18 labeled radioligands for detection of MAO-B activity

The aim of this project was to synthesize and evaluate three novel fluorine-18 labeled derivatives of propargyl amine as potential PET radioligands to visualize monoamine oxidase B (MAO-B) activity.The three fluorinated derivatives of propargyl amine ((S)-1-fluoro-N,4-dimethyl-N-(prop-2-ynyl)-pent-4-en-2-amine (5), (S)-N-(1-fluoro-3-(furan-2-yl)propan-2-yl)-N-methylprop-2-yn-1-amine (10) and (S)-1-fluoro-N,4-dimethyl-N-(prop-2-ynyl)pentan-2-amine (15)) were synthesized in multi-step organic syntheses. IC₅₀ values for inhibition were determined for compounds 5, 10 and 15 in order to determine their specificity for binding to MAO-B. Compound 5 inhibited MAO-B with an IC₅₀ of 664 ± 48.08 nM. No further investigation was carried out with this compound. Compound 10 inhibited MAO-B with an IC₅₀ of 208.5 ± 13.44 nM and compound 15 featured an IC₅₀ of 131.5 ± 0.71 nM for its MAO-B inhibitory activity. None of the compounds inhibited MAO-A activity (IC₅₀ > 2 μM).The fluorine-18 labeled analogues of the two higher binding affinity compounds (10 and 15) (S)-N-(1-[¹⁸F]fluoro-3-(furan-2-yl)propan-2-yl)-N-methylprop-2-yn-1-amine (16) and (S)-1-[¹⁸F]fluoro-N,4-dimethyl-N-(prop-2-ynyl)pentan-2-amine (18) were both prepared from the corresponding precursors 9A, 9B and 14A, 14B by a one-step fluorine-18 nucleophilic substitution reaction. Autoradiography experiments on human postmortem brain tissue sections were performed with 16 and 18. Only compound 18 demonstrated a high selectivity for MAO-B over MAO-A and was, therefore, chosen for further examination by PET in a cynomolgus monkey.The initial uptake of 18 in the monkey brain was 250% SUV at 4 min post injection. The highest uptake of radioactivity was observed in the striatum and thalamus, regions with high MAO-B activity, whereas lower levels of radioactivity were detected in the cortex and cerebellum. The percentage of unchanged radioligand 18 was 30% in plasma at 90 min post injection.In conclusion, compound 18 is a selective inhibitor of MAO-B in vitro and demonstrated a MAO-B specific binding pattern in vivo by PET in monkey. It can, therefore, be considered as a candidate for further investigation in human by PET.     

Spatial patterns of mayfly,stonefly and caddisfly assemblages in Swiss running waters in the face of global warming

Biotic homogenization represents a major concern in ecology but relatively few studies have assessed climate change impacts on assemblage patterns of freshwater species. Our main goals were to predict the current and future (years 2035, 2060 and 2085) patterns of mayfly, stonefly and caddisfly (EPT) diversity across Switzerland from macroscale environmental variables, and to assess the impact of warming temperatures on β-diversity. The study area was the entire Swiss territory divided into 21 818 subcatchments (median area of 1.41 km²), used as spatial units for predicting patterns of EPT diversity. We assumed that the stream conditions were homogeneous within a subcatchment at this scale. Incidence of EPT larvae was derived from samplings carried out between 2010 and 2017 in 292 water-course sites as part of two national monitoring programs. We employed generalized dissimilarity modeling to analyze the spatial turnover of EPT assemblages. Climatic, topographic, geological and land-use variables were used as covariates, and different climate change scenarios were used for future predictions. We compared β-diversity among the different scenarios through distance-based tests of homogeneity of multivariate dispersions. Our findings showed the largest amount of EPT turnover occurred along the air temperature and slope gradients, considered as good proxies for water temperature and flow velocity. We predicted a biotic homogenization with increasing temperatures due to the upstream expansion of some species from the sub-montane level, which only stabilizes in the most conservative climate scenario. This study is the first countrywide prediction of EPT composition patterns in the context of global warming and provides insights into the vulnerability of assemblages occurring at high elevations.     

More widespread alien tree species do not have larger impacts on regeneration of native tree species in a tropical forest reserve

There is insufficient information regarding the factors affecting the environmental impacts of alien species. In particular, little is known about whether there is any relationship between the invasiveness (establishment and spread) of an introduced species and its per capita impact. We experimentally assessed the relationship between the extent of spread of up to 29 alien plant species and their impact on recruitment of native tree species in Amani Botanical Garden, Tanzania. We also studied the effects of allelochemicals of selected alien on native plant species to assess potential mechanisms of impact. We found no relationship between the extent of spread of an alien tree species and their impact on seed germination, seedling survival, and seedling communities of native trees in their understory, and no indication that allelochemicals consistently explain their effects on recruitment of the studied species. These results suggest that extent of spread cannot be used as a proxy for impact. Hence, managers should continue assessing both the spread and the impact of alien species when prioritizing alien species for management.     

Runoff sources and land cover change in the Amazon: an end-member mixing analysis from small watersheds

The flowpaths by which water moves from watersheds to streams has important consequences for the runoff dynamics and biogeochemistry of surface waters in the Amazon Basin. The clearing of Amazon forest to cattle pasture has the potential to change runoff sources to streams by shifting runoff to more surficial flow pathways. We applied end-member mixing analysis (EMMA) to 10 small watersheds throughout the Amazon in which solute composition of streamwater and groundwater, overland flow, soil solution, throughfall and rainwater were measured, largely as part of the Large-Scale Biosphere-Atmosphere Experiment in Amazonia. We found a range in the extent to which streamwater samples fell within the mixing space determined by potential flowpath end-members, suggesting that some water sources to streams were not sampled. The contribution of overland flow as a source of stream flow was greater in pasture watersheds than in forest watersheds of comparable size. Increases in overland flow contribution to pasture streams ranged in some cases from 0% in forest to 27?C28% in pasture and were broadly consistent with results from hydrometric sampling of Amazon forest and pasture watersheds that indicate 17- to 18-fold increase in the overland flow contribution to stream flow in pastures. In forest, overland flow was an important contribution to stream flow (45?C57%) in ephemeral streams where flows were dominated by stormflow. Overland flow contribution to stream flow decreased in importance with increasing watershed area, from 21 to 57% in forest and 60?C89% in pasture watersheds of less than 10?ha to 0% in forest and 27?C28% in pastures in watersheds greater than 100?ha. Soil solution contributions to stream flow were similar across watershed area and groundwater inputs generally increased in proportion to decreases in overland flow. Application of EMMA across multiple watersheds indicated patterns across gradients of stream size and land cover that were consistent with patterns determined by detailed hydrometric sampling.     

Baboon (Papio anubis) social complexity--a network approach

Although many studies have analyzed the causes and consequences of social relationships, few studies have explicitly assessed how measures of social relationships are affected by the choice of behaviors used to quantify them. The use of many behaviors to measure social relationships in primates has long been advocated, but it was analytically difficult to implement this framework into primatological work. However, recent advances in social network analysis (SNA) now allow the comparison of multiple networks created from different behaviors. Here we use our database of baboon social behavior (Papio anubis, Gashaka Gumti National Park, Nigeria) to investigate (i) to what extent social networks created from different behaviors overlap, (ii) to what extent individuals occupy similar social positions in these networks and (iii) how sex affects social network position in this population of baboons. We used data on grooming, aggression, displacement, mounting and presenting, which were collected over a 15-month period. We calculated network parameters separately for each behavior. Networks based on displacement, mounting and presenting were very similar to each other, whereas grooming and aggression networks differed both from each other and from mounting, displacement and presenting networks. Overall, individual network positions were strongly affected by sex. Individuals central in one network tended to be central in most other networks as well, whereas other measures such as clustering coefficient were found to vary depending on the behavior analyzed. Thus, our results suggest that a baboon's social environment is best described by a multiplex network based on affiliative, aggressive and sexual behavior. Modern SNA provides a number of useful tools that will help us to better understand animals' social environment. We also discuss potential caveats related to their use.     

High-resolution proxies for wood density variations in Terminalia superba

Background and Aims

Density is a crucial variable in forest and wood science and is evaluated by a multitude of methods. Direct gravimetric methods are mostly destructive and time-consuming. Therefore, faster and semi- to non-destructive indirect methods have been developed.

Methods

Profiles of wood density variations with a resolution of approx. 50 µm were derived from one-dimensional resistance drillings, two-dimensional neutron scans, and three-dimensional neutron and X-ray scans. All methods were applied on Terminalia superba Engl. & Diels, an African pioneer species which sometimes exhibits a brown heart (limba noir).

Key Results

The use of X-ray tomography combined with a reference material permitted direct estimates of wood density. These X-ray-derived densities overestimated gravimetrically determined densities non-significantly and showed high correlation (linear regression, R² = 0·995). When comparing X-ray densities with the attenuation coefficients of neutron scans and the amplitude of drilling resistance, a significant linear relation was found with the neutron attenuation coefficient (R² = 0·986) yet a weak relation with drilling resistance (R² = 0·243). When density patterns are compared, all three methods are capable of revealing the same trends. Differences are mainly due to the orientation of tree rings and the different characteristics of the indirect methods.

Conclusions

High-resolution X-ray computed tomography is a promising technique for research on wood cores and will be explored further on other temperate and tropical species. Further study on limba noir is necessary to reveal the causes of density variations and to determine how resistance drillings can be further refined.     

Effects of chronic weight perturbation on energy homeostasis and brain structure in mice

Maintenance of reduced body weight in lean and obese human subjects results in the persistent decrease in energy expenditure below what can be accounted for by changes in body mass and composition. Genetic and developmental factors may determine a central nervous system (CNS)-mediated minimum threshold of somatic energy stores below which behavioral and metabolic compensations for weight loss are invoked. A critical question is whether this threshold can be altered by environmental influences and by what mechanisms such alterations might be achieved. We examined the bioenergetic, behavioral, and CNS structural responses to weight reduction of diet-induced obese (DIO) and never-obese (CON) C57BL/6J male mice. We found that weight-reduced (WR) DIO-WR and CON-WR animals showed reductions in energy expenditure, adjusted for body mass and composition, comparable (-10-15%) to those seen in human subjects. The proportion of excitatory synapses on arcuate nucleus proopiomelanocortin neurons was decreased by ～50% in both DIO-WR and CON-WR mice. These data suggest that prolonged maintenance of an elevated body weight (fat) alters energy homeostatic systems to defend a higher level of body fat. The synaptic changes could provide a neural substrate for the disproportionate decline in energy expenditure in weight-reduced individuals. This response to chronic weight elevation may also occur in humans. The mouse model described here could help to identify the molecular/cellular mechanisms underlying both the defense mechanisms against sustained weight loss and the upward resetting of those mechanisms following sustained weight gain.     

OxLDL-mediated survival of macrophages does not require LDL internalization or signalling by major pattern recognition receptors

Macrophages play a key role in the pathogenesis of atherosclerosis, in part by destabilizing plaques. We and others have shown that low concentrations of oxidized LDL (oxLDL) inhibit macrophage apoptosis. As oxLDL is present in lesions, this may be a mechanism by which macrophage populations in the intima are expanded. We have previously shown that oxLDL activates prosurvival signalling pathways such as the phosphoinositide 3-kinase (PI3K) pathway in bone marrow derived macrophages (BMDMs). However, little is known about more upstream signalling events especially at the receptor level. The endocytic pattern recognition receptors (PRRs), scavenger receptor A (SR-A) and CD36, are the main receptors on macrophages for uptake of oxLDL and are therefore important in foam cell formation. The signalling PRRs such as toll-like receptor (TLR) 2 and 4 also bind some types of oxLDL. This study was done to determine if any of the known PRRs are required for the anti-apoptotic effects of oxLDL in BMDMs. To do this, we tested the effect of oxLDL on viability of BMDMs lacking both SR-A and CD36 or lacking TLR2, TLR4, CD14, FcγRIIb, or RAGE. Our results indicate that none of these receptors are essential for activating the oxLDL prosurvival pathway. Furthermore, we show that the anti-apoptotic effect is not dependent on the uptake of oxLDL.     

Impact of arbuscular mycorrhizal fungi on the allergenic potential of tomato

Arbuscular mycorrhizal (AM) fungi influence the expression of defence-related genes in roots and can cause systemic resistance in plants probably due to the induced expression of specific defence proteins. Among the different groups of defence proteins, plant food allergens were identified. We hypothesized that tomato-allergic patients differently react to tomatoes derived from plants inoculated or not by mycorrhizal fungi. To test this, two tomato genotypes, wild-type 76R and a nearly isogenic mycorrhizal mutant RMC, were inoculated with the AM fungus Glomus mosseae or not under conditions similar to horticultural practice. Under such conditions, the AM fungus showed only a very low colonisation rate, but still was able to increase shoot growth of the wild-type 76R. Nearly no colonisation was observed in the mutant RMC, and shoot development was also not affected. Root fresh weights were diminished in AM-inoculated plants of both genotypes compared to the corresponding controls. No mycorrhizal effects were observed on the biomass and the concentration of phosphate and nitrogen in fruits. Real-time quantitative polymerase chain reaction analysis revealed that six among eight genes encoding for putative allergens showed a significant induced RNA accumulation in fruits of AM-colonised plants. However, human skin reactivity tests using mixed samples of tomato fruits from the AM-inoculated and control plants showed no differences. Our data indicate that AM colonisation under conditions close to horticultural practice can induce the expression of allergen-encoding genes in fruits, but this does not lead necessarily to a higher allergenic potential.