Investigations into the in vitro developmental plasticity of adult mesenchymal stem cells

Bone marrow (BM) derived stem cells contribute to the regeneration of diverse adult tissues including heart, liver and brain following BM transplantation. Trans- differentiation is a mechanism proposed to explain how tissue specific stem cells could generate cells of other organs, thus supporting the emerging concept of enhanced adult stem cell plasticity. New studies have demonstrated that spontaneous cell fusion rather than trans-differentiation is the cause of unexpected cell fate changes in vivo. In contrast, several authors have reported that trans-differentiation can occur in vitro in the absence of cell fusion, including the generation of neural derivatives from non-neural tissues. These findings have profound implications for stem cell biology and cell replacement therapy, and as a result require extensive validation. Mesenchymal stem cells (MSCs) nave been isolated from the postnatal BM and more recently many other sites including adipose tissue, skin and placental cord blood. As such these cells have attracted interest as candidates for cell replacement therapies. This interest follows recent observations both in vitro and in transplant studies that these cells are capable of broader differentiation potential beyond those cell lineages associated with the organ in which they reside. The aim of the present thesis was to examine the developmental plasticity of MSCs in vitro including the capacity of these cells to cross lineage boundaries by differentiating into neuro-ectodermal cell derivatives. There are no universally accepted procedures for the prospective isolation of these cells. In the present thesis, procedures for the isolation of MSCs from rat BM and optimal conditions for the propagation of these cells in culture without loss of multipotent differentiation potential and proliferative capacity are first described. Secondly, the response of cultured MSCs with a consistent immunophenotype to defined culture conditions, previously reported to induce neuronal differentiation of MSCs are evaluated. Thirdly, evidence is presented that suggests that previous claims of trans-differentiation and apparent changes in cell phenotype have been incorrectly interpreted. Evidence is provided that MSCs respond to neural cues in vitro with a stress response, which is characterized by aberrant changes in the expression of constitutive neural proteins, an event previously interpreted as trans-differentiation. MSCs do not have the attributes of early or mature neural derivatives and therefore such changes in protein expression do not equate to true neural differentiation. Finally, evidence is presented that demonstrates that MSCs cultured under defined culture conditions release soluble factors that instruct a neurogenic cell fate decision on neural stem cells (NSCs). In addition, these soluble factors also increase neurite outgrowth of Tuj-1+ differentiating cell progeny. These effects may in part explain the therapeutic benefit of MSG transplantation in animal models of CNS lesions

Male sexually coercive behaviour drives increased swimming efficiency in female guppies

Sexual coercion of females by males is widespread across sexually reproducing species. It stems from a conflict of interest over reproduction and exerts selective pressure on both sexes. For females, there is often a significant energetic cost of exposure to male sexually coercive behaviours. Our understanding of the efficiency of female resistance to male sexually coercive behaviour is key to understanding how sexual conflict contributes to population level dynamics and ultimately to the evolution of sexually antagonistic traits. Overlooked within this context are plastic physiological responses of traits within the lifetime of females that could moderate the energetic cost imposed by coercive males. Here, we examined whether conflict over the frequency and timing of mating between male and female guppies Poecilia reticulata can induce changes in swimming performance and aerobic capacity in females as they work to escape harassment by males. Females exposed to higher levels of harassment over a 5-month period used less oxygen to swim at a given speed, but displayed no difference in resting metabolic rate, maximal metabolic rate, maximal sustained swimming speed or aerobic scope compared to females receiving lower levels of harassment. The observed increase in swimming efficiency is at least partially related to differences in swimming mechanics, likely brought on by a training effect of increased activity, as highly harassed females spent less time performing pectoral fin-assisted swimming. Sexual conflict results in sexually antagonistic traits that impose a variety of costs, but our results show that females can reduce costs through phenotypic plasticity. It is also possible that phenotypic plasticity in swimming physiology or mechanics in response to sexual coercion can potentially give females more control over matings and affect which male traits are under selection

On the Search for Quasar Light Echoes

The UV radiation from a quasar leaves a characteristic pattern in the distribution of ionized hydrogen throughout the surrounding space. This pattern or light echo propagates through the intergalactic medium at the speed of light, and can be observed by its imprint on the Ly-alpha forest spectra of background sources. As the echo persists after the quasar has switched off, it offers the possibility of searching for dead quasars, and constraining their luminosities and lifetimes. We outline a technique to search for and characterize these light echoes. To test the method, we create artificial Ly-alpha forest spectra from cosmological simulations at z=3, apply light echoes and search for them. We show how the simulations can also be used to quantify the significance level of any detection. We find that light echoes from the brightest quasars could be found in observational data. With absorption line spectra of 100 redshift z~3-3.5 quasars or galaxies in a 1 square degree area, we expect that ~10 echoes from quasars with B band luminosities L_B=3x10^45 ergs/s exist that could be found at 95% confidence, assuming a quasar lifetime of ~10^7 yr. Even a null result from such a search would have interesting implications for our understanding of quasar luminosities and lifetimes.Comment: 9 pages, 7 figures, ApJ in pres

Physical effects on the Lyman-alpha forest flux power spectrum: damping wings, ionizing radiation fluctuations, and galactic winds

We explore several physical effects on the power spectrum of the Lyman-alpha forest transmitted flux. The effects we investigate here are usually not part of hydrodynamic simulations and so need to be estimated separately. The most important effect is that of high column density absorbers with damping wings, which add power on large scales. We compute their effect using the observational constraints on their abundance as a function of column density. Ignoring their effect leads to an underestimation of the slope of the linear theory power spectrum. The second effect we investigate is that of fluctuations in the ionizing radiation field. For this purpose we use a very large high resolution N-body simulation, which allows us to simulate both the fluctuations in the ionizing radiation and the small scale LyaF within the same simulation. We find an enhancement of power on large scales for quasars and a suppression for galaxies. The strength of the effect rapidly increases with increasing redshift, allowing it to be uniquely identified in cases where it is significant. We develop templates which can be used to search for this effect as a function of quasar lifetime, quasar luminosity function, and attenuation length. Finally, we explore the effects of galactic winds using hydrodynamic simulations. We find the wind effects on the LyaF power spectrum to be be degenerate with parameters related to the temperature of the gas that are already marginalized over in cosmological fits. While more work is needed to conclusively exclude all possible systematic errors, our results suggest that, in the context of data analysis procedures where parameters of the LyaF model are properly marginalized over, the flux power spectrum is a reliable tracer of cosmological information.Comment: 13 pages, 8 figures, to be submitted to MNRA

Ionizing radiation fluctuations and large-scale structure in the Lyman-alpha forest

We investigate the large-scale inhomogeneities of the hydrogen ionizing radiation field in the Universe at redshift z=3. Using a raytracing algorithm, we simulate a model in which quasars are the dominant sources of radiation. We make use of large scale N-body simulations of a LambdaCDM universe, and include such effects as finite quasar lifetimes and output on the lightcone, which affects the shape of quasar light echoes. We create Lya forest spectra that would be generated in the presence of such a fluctuating radiation field, finding that the power spectrum of the Lya forest can be suppressed by as much as 15 % for modes with k=0.05-1 Mpc/h. This relatively small effect may have consequences for high precision measurements of the Lya power spectrum on larger scales than have yet been published. We also investigate another radiation field probe, the cross-correlation of quasar positions and the Lya forest. For both quasar lifetimes which we simulate (10^7 yr and 10^8 yr), we expect to see a strong decrease in the Lya absorption close to other quasars (the ``foreground'' proximity effect). We then use data from the Sloan Digital Sky Survey First Data Release to make an observational determination of this statistic. We find no sign of our predicted lack of absorption, but instead increased absorption close to quasars. If the bursts of radiation from quasars last on average < 10^6 yr, then we would not expect to be able to see the foreground effect. However, the strength of the absorption itself seems to be indicative of rare objects, and hence much longer total times of emission per quasar. Variability of quasars in bursts with timescales > 10^4yr and < 10^6 yr could reconcile these two facts.Comment: Submitted to ApJ, 21 pages, 17 postscript figures, emulateapj.st

Socializing on MOOCs: comparing university and self-enrolled students

International audienceMOOCs are becoming more and more integrated in the higher education landscape of learning, with many institutions now pushing their students towards MOOC as part of their curriculum. But what does it mean for other MOOC learners? Are these students socializing the same way when they have an easier possibility to interact with classmates offline? Is the fact that they do not personally choose to enroll in a MOOC also having an effect? In this paper, we compare university-enrolled students to other MOOC participants and in particular other self-enrolled students, to examine how and why they socialize on and around the MOOC. Using data from two French MOOCs in project management, we show that university-enrolled students are less attracted by forums and seem to interact less than others when the workload increases , which could lead to misleading conclusions when analyzing data. We therefore encourage MOOC researchers to be particularly mindful of this new trend when performing social network analyses

Cellular mRNAs access second ORFs using a novel amino acid sequence-dependent coupled translation termination-reinitiation mechanism

Polycistronic transcripts are considered rare in the human genome. Initiation of translation of internal ORFs of eukaryotic genes has been shown to use either leaky scanning or highly structured IRES regions to access initiation codons. Studies on mammalian viruses identified a mechanism of coupled translation termination-reinitiation that allows translation of an additional ORF. Here, the ribosome terminating translation of ORF-1 translocates upstream to reinitiate translation of ORF-2. We have devised an algorithm to identify mRNAs in the human transcriptome in which the major ORF-1 overlaps a second ORF capable of encoding a product of at least 50 aa in length. This identified 4368 transcripts representing 2214 genes. We investigated 24 transcripts, 22 of which were shown to express a protein from ORF-2 highlighting that 3' UTRs contain protein-coding potential more frequently than previously suspected. Five transcripts accessed ORF-2 using a process of coupled translation termination-reinitiation. Analysis of one transcript, encoding the CASQ2 protein, showed that the mechanism by which the coupling process of the cellular mRNAs was achieved was novel. This process was not directed by the mRNA sequence but required an aspartate-rich repeat region at the carboxyl terminus of the terminating ORF-1 protein. Introduction of wobble mutations for the aspartate codon had no effect, whereas replacing aspartate for glutamate repeats eliminated translational coupling. This is the first description of a coordinated expression of two proteins from cellular mRNAs using a coupled translation termination-reinitiation process and is the first example of such a process being determined at the amino acid level

Defining and mapping the person with osteoarthritis for population studies and public health

OBJECTIVE: To determine population-based estimates for the prevalence of the person with OA, predicted to be the single greatest cause of disability in the general population by 2030, in order to inform the planning and commissioning of health, social care and prevention services. METHODS: A postal survey to all adults ≥50 years of age registered with eight general practices in the UK. Self-reported data on chronic joint pain in four body regions (hand, hip, knee, foot) and the disabling nature of the pain was collected to determine gender and age-group specific prevalence estimates of clinical OA in the joint region and in the person. Multiple imputation and weighted logistic regression was used to allow for missing data. RESULTS: A total of 26 705 mailed surveys resulted in 18 474 responses (adjusted response = 71.8%). Approximately half of the mailed population had OA in at least one of the four regions (53.23%, 95% CI 52.3, 54.1) and less than half of these had disabling OA (21.87%, 95% CI 21.2, 22.5). The more joint regions involved, the more likely that the OA was disabling. OA prevalence was higher in females and increased with age. Applied to the population of England, this yielded an estimated 3.5 million persons with disabling OA, including 1.45 million people between 50 and 65 years of age and 370 000 ≥85 years of age. CONCLUSIONS: A simple approach to defining the person with OA can contribute to population comparisons, public health projections and health care needs assessments

No Evidence for Orbital Loop Currents in Charge Ordered YBa2_2Cu3_3O6+x_{6+x} from Polarized Neutron Diffraction

It has been proposed that the pseudogap state of underdoped cuprate superconductors may be due to a transition to a phase which has circulating currents within each unit cell. Here, we use polarized neutron diffraction to search for the corresponding orbital moments in two samples of underdoped YBa$_2$Cu$_3$O$_{6+x}$ with doping levels $p=0.104$ and 0.123. In contrast to some other reports using polarized neutrons, but in agreement with nuclear magnetic resonance and muon spin rotation measurements, we find no evidence for the appearance of magnetic order below 300 K. Thus, our experiment suggests that such order is not an intrinsic property of high-quality cuprate superconductor single crystals. Our results provide an upper bound for a possible orbital loop moment which depends on the pattern of currents within the unit cell. For example, for the CC-$\theta_{II}$ pattern proposed by Varma, we find that the ordered moment per current loop is less than 0.013 $\mu_B$ for $p=0.104$.Comment: Comments in arXiv:1710.08173v1 fully addresse