2-Arylbenzoxazoles as novel cholesteryl ester transfer protein inhibitors: optimization via array synthesis

2-Arylbenzoxazole 5 was identified as a hit from a fluorescence-based high-throughput screen for CETP inhibitors. The synthesis and SAR investigation employing array synthesis of the A- and B-rings are described.     

Speciation in the highlands of Mexico: genetic and phenotypic divergence in the Mexican jay (Aphelocoma ultramarina)

The pine-oak woodlands of the Mexican highlands harbour significant biological diversity, yet little is known about the evolutionary history of organisms inhabiting this region. We assessed genetic and phenotypic differentiation in 482 individuals representing 27 populations of the Mexican jay ( Aphelocoma ultramarina ) — a widespread bird species of the Mexican highlands — to test whether populations in the central and northern Mexican sierras display discrete breaks between groups, which would be consistent with a role for the different mountain chains in divergence and speciation. We found abrupt breaks in mitochondrial DNA (mtDNA; ND2 and control region) delineating four major genetic groups found in the Sierra Madre Occidental, Sierra Madre Oriental, southern Central Plateau (Bajio), and Transvolcanic Belt. These mtDNA groups were largely corroborated by data from nuclear microsatellites and phenotypic data, except that clades from the Central Plateau and Sierra Madre Oriental showed clinal change in these data sets. Uncertainty about the mutation rate for our mitochondrial markers warrants considerable caution with regard to estimating divergence times, but the major genetic groups appear to have split before the most extreme period of glacial cycling that marked the last 0.7 million years and after Mexico's period of major mountain formation. The fact that some genetic breaks do not coincide with well-known geographic barriers suggests a role for ecology in divergence and speciation, and we discuss implications for taxonomy and conservation.     

Rapid mass spectrometric analysis of 15N-Leu incorporation fidelity during preparation of specifically labeled NMR samples

Advances in NMR spectroscopy have enabled the study of larger proteins that typically have significant overlap in their spectra. Specific (15)N-amino acid incorporation is a powerful tool for reducing spectral overlap and attaining reliable sequential assignments. However, scrambling of the label during protein expression is a common problem. We describe a rapid method to evaluate the fidelity of specific (15)N-amino acid incorporation. The selectively labeled protein is proteolyzed, and the resulting peptides are analyzed using MALDI mass spectrometry. The (15)N incorporation is determined by analyzing the isotopic abundance of the peptides in the mass spectra using the program DEX. This analysis determined that expression with a 10-fold excess of unlabeled amino acids relative to the (15)N-amino acid prevents the scrambling of the (15)N label that is observed when equimolar amounts are used. MALDI TOF-TOF MS/MS data provide additional information that shows where the "extra" (15)N labels are incorporated, which can be useful in confirming ambiguous assignments. The described procedure provides a rapid technique to monitor the fidelity of selective labeling that does not require a lot of protein. These advantages make it an ideal way of determining optimal expression conditions for selectively labeled NMR samples.     

The Ediacaran emergence of bilaterians: congruence between the genetic and the geological fossil records

Unravelling the timing of the metazoan radiation is crucial for elucidating the macroevolutionary processes associated with the Cambrian explosion. Because estimates of metazoan divergence times derived from molecular clocks range from quite shallow (Ediacaran) to very deep (Mesoproterozoic), it has been difficult to ascertain whether there is concordance or quite dramatic discordance between the genetic and geological fossil records. Here, we show using a range of molecular clock methods that the major pulse of metazoan divergence times was during the Ediacaran, which is consistent with a synoptic reading of the Ediacaran macrobiota. These estimates are robust to changes in priors, and are returned with or without the inclusion of a palaeontologically derived maximal calibration point. Therefore, the two historical records of life both suggest that although the cradle of Metazoa lies in the Cryogenian, and despite the explosion of ecology that occurs in the Cambrian, it is the emergence of bilaterian taxa in the Ediacaran that sets the tempo and mode of macroevolution for the remainder of geological time.     

Short-term behavioural response of polar bears (<Emphasis Type="Italic">Ursus maritimus</Emphasis>) to snowmobile disturbance

In this study the distance, at which polar bears detected and actively responded to approaching snowmobiles was measured and the behavioural response was recorded. The study was performed on Svalbard, an arctic island where human traffic has increased substantially in recent years. Fieldwork was conduced in April and/or May during the years 2003–2005. Polar bears were observed on ice with telescopes and binoculars. Undisturbed polar bears were observed continuously and their behaviours recorded, during the time when two snowmobiles moved toward the bear(s). Distances between the bear, the observer, and the approaching snowmobiles were measured using GPS positions taken on the track towards the bear. Data on the behavioural response of 20 encounters with bears were collected. On average, bears were alerted to the snowmobiles at 1,164 m. Mean distance at which the locomotive response occurred was 843 m, and there was a statistical significant difference in distance between sex and age classes [326 m (95% CI = 138–496 m) for adult males; 1,534 m (95% CI = 508–2,768 m) for adult females with cubs; 164 m (95% CI = 49–543 m) for two adult females without cubs; and 1,160 m (95% CI = 375–1,353 m) for single medium sized bears]. The responses of the polar bears to the snowmobiles were categorized according to intensity and persistence of reactions. Females with cubs and single medium sized bears tended to show more intense responses than adult males and lone adult females. Wind direction affects sound and odour transmission, and although an effect on response distance was not found, the response intensity was affected by wind direction. We conclude that female polar bears with small cubs in particular may have a greater risk to be disturbed, since they react at greater distances with amplified reactions; thus, users of snowmobiles should take particular care in areas where females with cubs are present.     

Movements of two Svalbard polar bears recorded using geographical positioning system satellite transmitters

Until recently, studies on polar bear (Ursus maritimus) movements and space use have used data collected by satellite telemetry collars that provided positions infrequently (typically weekly) and with low precision (by Doppler Shift method). A new generation of transmitters incorporated into collars use the Global Positioning System (GPS) to provide highly accurate positions, and have the ability to provide many positions per day. We used data from two GPS collars fitted to female polar bears, that attempted to collect six positions per day (4-h apart) for 546 days (from April 2000 to September 2001) and 413 days (from April 2000 to May 2001) to estimate how estimated speed of movement and home range size increase with increasing number of data points. Using all the positions, we estimated that the bears moved a minimum of 14.3 and 15.8 km per day on average. The fractal dimension (D) of the movement pathways for the two bears were D = 1.28 and 1.31, respectively, indicating low tortousity of the movements. Their minimum estimated annual home range areas were 20,794 and 112,183 km². Simulations showed that a commonly used sampling regime of one location every 6th day would have significantly underestimated the movement rates and the home range sizes compared to our estimates. We also used the high accuracy of GPS positions to look at distances moved within 4-h periods. Maximum movement rate during a 4-h period for the two bears was 4.21 and 4.58 km/h, respectively. Variation in median values by month was significant (0.01 km/h in November for N23476 to 1.48 km/h in December for N7955). Diurnal variation was observed to differ between defined periods.     

Selection on amino acid substitutions in Arabidopsis

Studies of nucleotide diversity have found an excess of low-frequency amino acid polymorphisms segregating in Arabidopsis thaliana, suggesting a predominance of weak purifying selection acting on amino acid polymorphism in this inbreeding species. Here, we investigate levels of diversity and divergence at synonymous and nonsynonymous sites in 6 circumpolar populations of the outbreeding Arabidopsis lyrata and compare these results with A. thaliana, to test for differences in mutation and selection parameters across genes, populations, and species. We find that A. lyrata shows an excess of low-frequency nonsynonymous polymorphisms both within populations and species wide, consistent with weak purifying selection similar to the patterns observed in A. thaliana. Furthermore, nonsynonymous polymorphisms tend to be more restricted in their population distribution in A. lyrata, consistent with purifying selection preventing their geographic spread. Highly expressed genes show a reduced ratio of amino acid to synonymous change for both polymorphism and fixed differences, suggesting a general pattern of stronger purifying selection on high-expression proteins.     

Genetic divergence of climatically marginal populations of Vicia pisiformis on the Scandinavian Peninsula

Vicia pisiformis L. is a perennial leguminous plant with a main distribution in broadleaved forest-steppes of eastern Europe. The species is classified as endangered (EN) according to the IUCN red-lists in both Norway and Sweden, due to severe fragmentation, small population sizes and continuing population decline. The populations on the Scandinavian Peninsula constitute the northern limit of the species distribution and are mostly restricted to warm stony slopes with predominantly southern aspects. In this study we used the AFLP method, which is a high-resolution genetic fingerprint method. Samples were collected from 22 Scandinavian populations. The overall genetic structure was analysed in an AMOVA, in a Mantel test and through constrained correspondence analysis (CCA). The ordination scores representing non-geographic genetic divergence were extracted from the CCA and analysed in a linear model using habitat variables and population size as explanatory variables. We found (i) a strong geographic structure, (ii) significant genetic divergence between populations, (iii) that this genetic divergence remained significant even after removing the effect of geography in a partial CCA and (iv) that the remaining non-geographic part of genetic divergence (distance from the ordination centre) was associated with aspect, populations with a northern aspect were more genetically divergent. Aspect explains more variation than population size and is the only variable retained in the minimal adequate model. We suggest that local adaptation has caused this divergence from an expected geographical pattern of genetic variation. This explanation is further supported by the association between aspect and specific AFLP fragments.     

Temporal Aroma Release Profile of Solid and Liquid Droplet Emulsions

The release kinetics of four model aroma compounds from coarse (d ₃₂ = 1.0 μm) and fine (d ₃₂ = 0.25 μm) eicosane and hydrogenated palm stearin (HPS) emulsions prepared with either solid or liquid lipid droplets were measured using a model mouth instrument. For both lipids, the release of aroma compounds from emulsions with solid droplets was higher than from emulsions with liquid droplets. This difference was greater for less polar aroma compounds. The release from solid eicosane droplets increased with particle size but no such effect was observed for HPS emulsions, however, the release from solid eicosane was higher than solid HPS. The initial aroma release profile of the solid droplet emulsion matches that of a similar liquid oil emulsion but requires much less added aroma. Meeting presentation: Presented at 98th AOCS Annual Meeting and Expo in Quebec City, Canada.