Localization of the genetic determinants of meiosis suppression in Daphnia pulex

Although approximately 1 in 10,000 animal species is capable of parthenogenetic reproduction, the evolutionary causes and consequences of such transitions remain uncertain. The microcrustacean Daphnia pulex provides a potentially powerful tool for investigating these issues because lineages that are obligately asexual in terms of female function can nevertheless transmit meiosis-suppressing genes to sexual populations via haploid sperm produced by environmentally induced males. The application of association mapping to a wide geographic collection of D. pulex clones suggests that sex-limited meiosis suppression in D. pulex has spread westward from a northeastern glacial refugium, conveyed by a dominant epistatic interaction among the products of at least four unlinked loci, with one entire chromosome being inherited through males in a nearly nonrecombining fashion. With the enormous set of genomic tools now available for D. pulex, these results set the stage for the determination of the functional underpinnings of the conversion of meiosis to a mitotic-like mode of inheritance.     

Estimating levels of inbreeding using AFLP markers

In the absence of detailed pedigree records, researchers have attempted to estimate individuals' levels of inbreeding using molecular markers, generally making use of heterozygosity measures based on microsatellite markers. Here we report and validate a method for estimating an individual's inbreeding coefficient, f, using amplified fragment length polymorphism (AFLP) markers. We use simulations to confirm that our measure scales appropriately with f when allele frequencies can be estimated from a subset of outbred individuals. We also present an approach for obtaining satisfactory estimates even in the absence of an independent set of known outbred individuals from which to estimate allele frequencies. We then test our method against empirical data from 179 wild and captive-bred old-field mice, Peromyscus polionotus subgriseus, comprising pedigree-based estimates of f, along with genetic data from 94 AFLP markers and 12 microsatellites. Inbreeding estimates based on both AFLP and microsatellite markers were found to correlate strongly with pedigree-based inbreeding coefficients. Owing to their ease of amplification in any species, AFLP markers may prove to be a valuable new tool for estimating f in natural populations and for examining correlations between heterozygosity and fitness.     

Effects of barley straw (Hordeum vulgare) on freshwater and brackish phytoplankton and cyanobacteria

A short-term laboratory study was conductedto investigate the effect of barley strawin controlling several common phytoplanktonand cyanobacterial species. Following aone-month incubation of barley straw incoarsely filtered fresh Potomac River andbrackish Patuxent River waters, the growthof six autotrophic taxa was followed inculture. Barley straw slurry reduced theyield of three taxa (Ankistrodesmusfalcatus, Chlorella capsulata, Isochrysis sp.) in comparison withcultures not receiving the slurry. Although no significant changes in growthwere detected with three other taxa (Cyclotella sp., Prorocentrumminimum, freshwater Pseudanabaenasp.), some patterns indicated potentialimpacts of the barley straw. First, ahigher addition of straw to Cyclotella sp. resulted in a lower biomassaccumulation than in cultures receivinglower levels. Second, the bloom-formingdinoflagellate Prorcentrum minimumwas apparently stimulated at low barleystraw levels, perhaps suggesting conditionsassociated with the straw(metals-chelation, bacterial-producednutrients) might stimulate dinoflagellategrowth. Third, species shifts wereobserved in two of the cultures, withbarley straw favoring shifts from Isochrysis to a Cyclotella sp. –Thalassiosira sp. mixture and shiftsfrom Pseudanabaena to a Pseudanabaena – Scenedesmus mixture. These results provide new records for thesusceptibility of freshwater and brackishphytoplankton taxa to barley strawexposure, including species-specificresponses and shifts in species dominancein mixed assemblages.     

Fetal Origins of Obesity

The worldwide epidemic of obesity continues unabated. Obesity is notoriously difficult to treat, and, thus, prevention is critical. A new paradigm for prevention, which evolved from the notion that environmental factors in utero may influence lifelong health, has emerged in recent years. A large number of epidemiological studies have demonstrated a direct relationship between birth weight and BMI attained in later life. Although the data are limited by lack of information on potential confounders, these associations seem robust. Possible mechanisms include lasting changes in proportions of fat and lean body mass, central nervous system appetite control, and pancreatic structure and function. Additionally, lower birth weight seems to be associated with later risk for central obesity, which also confers increased cardiovascular risk. This association may be mediated through changes in the hypothalamic pituitary axis, insulin secretion and sensing, and vascular responsiveness. The combination of lower birth weight and higher attained BMI is most strongly associated with later disease risk. We are faced with the seeming paradox of increased adiposity at both ends of the birth weight spectrum—higher BMI with higher birth weight and increased central obesity with lower birth weight. Future research on molecular genetics, intrauterine growth, growth trajectories after birth, and relationships of fat and lean mass will elucidate relationships between early life experiences and later body proportions. Prevention of obesity starting in childhood is critical and can have lifelong, perhaps multigenerational, impact.     

Anchorage-independent growth of breast carcinoma cells is mediated by serum exosomes

We hereby report studies that suggest a role for serum exosomes in the anchorage-independent growth (AIG) of tumor cells. In AIG assays, fetal bovine serum is one of the critical ingredients. We therefore purified exosomes from fetal bovine serum and examined their potential to promote growth of breast carcinoma cells in soft agar and Matrigel after reconstituting them into growth medium (EEM). In all the assays, viable colonies were formed only in the presence of exosomes. Some of the exosomal proteins we identified, have been documented by others and could be considered exosomal markers. Labeled purified exosomes were up-taken by the tumor cells, a process that could be competed out with excess unlabeled vesicles. Our data also suggested that once endocytosed by a cell, the exosomes could be recycled back to the conditioned medium from where they can be up-taken by other cells. We also demonstrated that low concentrations of exosomes activate MAP kinases, suggesting a mechanism by which they maintain the growth of the tumor cells in soft agar. Taken together, our data demonstrate that serum exosomes form a growth promoting platform for AIG of tumor cells and may open a new vista into cancer cell growth in vivo.     

Quantitative identification of teratoma tissues formed by human embryonic stem cells with TeratomEye

An automated vision system, TeratomEye, was developed for the identification of three representative tissue types: muscle, gut and neural epithelia which are commonly found in teratomas formed from human embryonic stem cells. Muscle tissue, a common structure was identified with an accuracy of 90.3% with high specificity and sensitivity greater than 90%. Gut epithelia were identified with an accuracy of 87.5% with specificity and sensitivity greater than 80%. Neural epithelia which were the most difficult structures to distinguish gave an accuracy of 47.6%. TeratomEye is therefore useful for the automated identification of differentiated tissues in teratoma sections.     

An application of the active time model to multiple concurrent variable-interval schedules

The current experiment investigates whether an active time model can account for anomalous results that have emerged from multiple schedule, concurrent variable-interval (VI) VI experiments. The model assumes that (1) during concurrent VI VI training pigeons learn a function that relates time since the most immediate response, i.e., active time, to changeover probabilities and (2) that molar preference is the result of an interaction between inter-response time frequencies and the learned active time changeover functions. Pigeons were trained under a concurrent VI 30-s VI 30-s schedule and a concurrent VI 60-s VI 60-s schedule. Probes were conducted in which VI 30-s and VI 60-s stimuli were paired. During these probes, birds allocated choices equally to the stimuli. The active time model accurately fit individual subject data. In contrast data were not fit by a variant of scalar expectancy theory proposed by Gibbon [Gibbon, J., 1995. Dynamics of time matching: arousal makes better seem worse. Psychon. Bull. Rev. 2, 208-215].