Comparative molecular field analysis and molecular dynamics studies of α/β hydrolase domain containing 6 (ABHD6) inhibitors

The endocannabinoid system remains an attractive molecular target for pharmacological intervention due to its roles in the central nervous system in learning, thinking, emotional function, regulation of food intake or pain sensation, as well as in the peripheral nervous system, where it modulates the action of cardiovascular, immune, metabolic or reproductive function. α/β hydrolase domain containing 6 (ABHD6)—an enzyme forming part of the endocannabinoid system—is a newly discovered post-genomic protein acting as a 2-AG (2-arachidonoylglycerol) serine hydrolase. We have recently reported a series of 1,2,5-thiadiazole carbamates as potent and selective ABHD6 inhibitors. Here, we present comparative molecular field analysis (CoMFA) and molecular dynamics studies of these compounds. First, we performed a homology modeling study of ABHD6 based on the assumption that the catalytic triad of ABHD6 comprises Ser148–His306–Asp 278 and the oxyanion hole is formed by Met149 and Phe80. A total of 42 compounds was docked to the homology model using the Glide module from the Schrödinger suite of software and the selected docking poses were used for CoMFA alignment. A model with the following statistics was obtained: R ² = 0.98, Q ² = 0.55. In order to study the molecular interactions of the inhibitors with ABHD6 in detail, molecular dynamics was performed with the Desmond program. It was found that, during the simulations, the hydrogen bond between the inhibitor carbonyl group and the main chain of Phe80 is weakened, whereas a new hydrogen bond with the side chain of Ser148 is formed, facilitating the possible formation of a covalent bond.

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Graphical Abstract Left–right: Docking pose of 1 in the binding pocket of α/β hydrolase domain containing 6 (ABHD6) selected for molecular alignment; CoMFA steric and electrostatic contour fields; changes in potential energy of the complex during simulations for the complex of 6 and ABHD6

     

A doubled haploid rye linkage map with a QTL affecting α-amylase activity

A rye doubled haploid (DH) mapping population (Amilo × Voima) segregating for pre-harvest sprouting (PHS) was generated through anther culture of F₁ plants. A linkage map was constructed using DHs, to our knowledge, for the first time in rye. The map was composed of 289 loci: amplified fragment length polymorphism (AFLP), microsatellite, random amplified polymorphic DNA (RAPD), retrotransposon-microsatellite amplified polymorphism (REMAP), inter-retrotransposon amplified polymorphism (IRAP), inter-simple sequence repeat (ISSR) and sequence-related amplified polymorphism (SRAP) markers, and extended altogether 732 cM (one locus in every 2.5 cM). All of the seven rye chromosomes and four unplaced groups were formed. Distorted segregation of markers (P ≤ 0.05) was detected on all chromosomes. One major quantitative trait locus (QTL) affecting α-amylase activity was found, which explained 16.1% of phenotypic variation. The QTL was localized on the long arm of chromosome 5R. Microsatellites SCM74, RMS1115, and SCM77, nearest to the QTL, can be used for marker-assisted selection as a part of a rye breeding program to decrease sprouting damage.     

Distribution of the major histocompatibility complex antigens on different cellular components of human liver

Monoclonal mouse antibodies to the "framework" determinants of the class I and II molecules of the major histocompatibility complex (MHC) were used to demonstrate the presence of the MHC antigens in human liver. First, the localization of these antigens was demonstrated from frozen section histology with indirect FITC immunofluorescence and the cell component(s) binding the mouse antibody were identified by rabbit marker antisera and indirect TRITC immunofluorescence. Second, the antigen expression on the cell surface was analyzed by the Staphylococcus aureus rosette method from cytological cell smears. All antibodies reacted with cells in the liver sinusoids, both with the Kupffer cells and at least partially with the sinusoidal endothelial cells. The same antisera reacted also with the bile duct cells, though weaker, and with some stromal cells in close proximity of the blood vessels. The vascular endothelial cells of hepatic artery, hepatic vein, and portal vein displayed no reaction. Thus human liver differs strikingly from, e.g., human kidney, where the vascular endothelial cells contain large amounts of MHC antigens on the cell surface. This difference may be one explanation to why liver allografts are less promptly rejected than renal allografts in man.     

Geographical patterns of species turnover in aquatic plant communities

1. A classic theory in biogeography predicts that high latitude communities are unstable. This may be because of decreased species richness or decreased environmental predictability and productivity towards the poles.
2. We studied latitudinal patterns in long-term community persistence of aquatic vascular plants in 112 Finnish lakes, situated within a 1000-km range from the northernmost to the southernmost lake.
3. Contrary to theoretical predictions, we found that the turnover rate of plant species in 45 years was inversely related to latitude. That is, plant communities in northern lakes were more persistent than communities in southern lakes. When we used multiple regression to find the best predictors of species turnover rate (TR), latitude was the only variable that was highly significantly related to species turnover rate. Area, species number, water transparency, pH and change in transparency did not notably explain the gradient observed.
4. The latitudinal trend was mainly because of lower species immigration rates at higher latitudes, whereas extinction rate did not so strongly decrease with increasing latitude. Immigrations and extinctions in the lakes were not in balance: the species numbers between the 1930s and 1980s increased more strongly in the southern than northern lakes.
5. We suggest that the inverse relationship between latitude and plant species TR in Finland is most probably caused by human influence on lakes, especially eutrophication and immigration of new species in southern latitudes. In addition, although species richness per lake did not decrease towards the north, the total species pool probably does, which means that in the north there are fewer species that can actually immigrate.     

No effects of mobile phone use on cortical auditory change‐detection in children: An ERP study

We investigated the effect of mobile phone use on the auditory sensory memory in children. Auditory event‐related potentials (ERPs), P1, N2, mismatch negativity (MMN), and P3a, were recorded from 17 children, aged 11–12 years, in the recently developed multi‐feature paradigm. This paradigm allows one to determine the neural change‐detection profile consisting of several different types of acoustic changes. During the recording, an ordinary GSM (Global System for Mobile Communications) mobile phone emitting 902 MHz (pulsed at 217 Hz) electromagnetic field (EMF) was placed on the ear, over the left or right temporal area (SAR_1g = 1.14 W/kg, SAR_10g = 0.82 W/kg, peak value = 1.21 W/kg). The EMF was either on or off in a single‐blind manner. We found that a short exposure (two 6 min blocks for each side) to mobile phone EMF has no statistically significant effects on the neural change‐detection profile measured with the MMN. Furthermore, the multi‐feature paradigm was shown to be well suited for studies of perception accuracy and sensory memory in children. However, it should be noted that the present study only had sufficient statistical power to detect a large effect size. Bioelectromagnetics 31:191–199, 2010. © 2009 Wiley‐Liss, Inc.     

Species immigration, extinction and turnover of vascular plants in boreal lakes

Dictated by limited resource availability for land acquisition, a central question in conservation biology is the ability of areas of different size to maintain species diversity. The selected reserves should not only be species rich at the moment, but should also maintain species diversity in the long run. We used two sets of data on vascular plant species in boreal lakes collected in 1933/34 and 1996 to test the relationships between lake area and the extinction, immigration and turnover rates of the species. Moreover, we investigated, whether the number of species in 1933/34 or water connection between lakes was related to extinction, immigration and turnover rates of species. We found that lake area or shoreline length was not correlated with immigration or turnover rate. But extinction rate was slightly negatively correlated with shoreline length. The original number of species was positively related to the number of species extinctions and to the absolute turnover rate in the lakes, which indicates that species richness does not create stability in these communities. Species number was not correlated with immigration rate. Upstream water connections in the lakes did not affect immigration, extinction or turnover rates. We conclude that length of the shoreline is a better measure of suitable area for water plants than the lake area, and that because the correlation between shoreline length and extinction rate was slight, also small lakes can be valuable for conservation.     

Effect of nutrition of oocyte donor on the outcomes of somatic cell nuclear transfer in the sheep

The purpose of this study was to determine if the nutrition of the oocyte donor ewe influenced the success of somatic cell cloning. Merino ewes were fed at either a high- or a low-nutrition level for 3-5 mo before superovulation treatments. Freshly ovulated oocytes were enucleated and fused with serum-starved adult granulosa cells, and resulting reconstructed embryos were cultured for 6 days in modified synthetic oviduct fluid. Embryo cleavage and development to blastocysts were recorded, and good-quality embryos were transferred to synchronized recipient ewes either fresh or, on a few occasions, after vitrification. Pregnancies were monitored by ultrasonography from Day 40 of pregnancy, and offspring were delivered by either cesarean section or vaginal delivery. No differences occurred in the numbers of follicles aspirated, of oocytes recovered, or of oocytes utilizable for cloning between the high and low groups. Neither were there treatment differences in development to the blastocyst stage. However, transfer of embryos from the high group led to significantly more pregnancies and implanted fetuses. Also, more of the established pregnancies from the high group were carried to term, although this difference was not statistically significant. Lamb mortality was high, with half the live-born perishing soon after birth and more succumbing to various infections within days or weeks of birth, but no clear association between the offspring fate and the treatment group could be established. These results suggest that more research into the effect of nutrition on oocyte quality and its subsequent effect on cloning is warranted.     

Circannual leptin and ghrelin levels of the blue fox (Alopex lagopus) in reference to seasonal rhythms of body mass, adiposity, and food intake

The aim of the study was to investigate the circannual rhythms of leptin and ghrelin in the blue fox, a variant of the endangered arctic fox, in relation to its seasonal cycles of body mass, adiposity and food intake. The effects of long-term fasting and exogenous melatonin treatment on these weight-regulatory hormones were also investigated. The leptin concentrations of the blue fox increased during the autumnal accumulation of fat and decreased during the wintertime and vernal weight loss periods. The leptin levels peaked 2-6 weeks before the maximum values were observed for the body mass indices, voluntary food intake, and body masses. The ghrelin concentrations fluctuated widely during the autumn but decreased in the winter in association with suppression of food intake. Exogenous melatonin advanced the seasonal changes in the food intake of the blue fox but did not affect the seasonal rhythms of leptin and ghrelin concentrations. The leptin concentrations did not respond to the 3-week fasting periods in a consistent way, but the ghrelin levels increased due to food deprivation. In addition to the amount of fat in the body the leptin secretion of the blue fox may be regulated also by other factors. The blue fox may also express seasonal changes in its leptin sensitivity. Our results reinforce the hypothesis that leptin does not function as an acute indicator of body adiposity in seasonal carnivores but rather as a long-term signal of nutritional status.     

Endocrine response to fasting in the overwintering captive raccoon dog (Nyctereutes procyonoides)

The raccoon dog (Nyctereutes procyonoides) is an omnivorous canid utilizing the passive wintering strategy in the boreal climate. Farmed raccoon dogs (n=12) were randomly assigned into two study groups on 26 November 2003. Between 3 December 2003 and 27 January 2004, half of the animals were fasted for 8 weeks and plasma weight-regulatory hormone concentrations determined on 26 November and 30 December 2003 and on 27 January 2004. The plasma peptide YY, ghrelin, and growth hormone (GH) concentrations increased due to food deprivation, while the T4 and Acrp30 concentrations decreased. Furthermore, the plasma GH concentrations were higher in the fasted raccoon dogs than in the fed animals, which had higher plasma insulin, glucagon, and T4 concentrations. However, fasting had no effect on the plasma leptin concentrations. The results confirm previous findings with unchanged leptin levels in fasting carnivores. Increased GH levels probably contribute to increased lipolysis and mobilization of fat stores. Ghrelin can also enhance lipolysis by increasing the GH levels. The decreased levels of T4 may reduce the metabolic rate. The plasma dopamine concentrations decreased due to fasting unlike observed previously in rats. Together with the unaffected adrenaline, noradrenaline, and cortisol concentrations, this suggests that food deprivation in winter does not cause stress to the raccoon dog but is an integral part of its natural life history.