Selective Visualization of Fluorescent Sterols in Caenorhabditis elegans by Bleach-Rate-Based Image Segmentation

The nematode Caenorhabditis elegans is a genetically tractable model organism to investigate sterol transport. In vivo imaging of the fluorescent sterol, dehydroergosterol (DHE), is challenged by C. elegans' high autofluorescence in the same spectral region as emission of DHE. We present a method to detect DHE selectively, based on its rapid bleaching kinetics compared to cellular autofluorescence. Worms were repeatedly imaged on an ultraviolet-sensitive wide field (UV-WF) microscope, and bleaching kinetics of DHE were fitted on a pixel-basis to mathematical models describing the intensity decay. Bleach-rate constants were determined for DHE in vivo and confirmed in model membranes. Using this method, we could detect enrichment of DHE in specific tissues like the nerve ring, the spermateca and oocytes. We confirm these results in C. elegans gut-granule-loss (glo) mutants with reduced autofluorescence and compare our method with three-photon excitation microscopy of sterol in selected tissues. Bleach-rate-based UV-WF imaging is a useful tool for genetic screening experiments on sterol transport, as exemplified by RNA interference against the rme-2 gene coding for the yolk receptor and for worm homologues of Niemann-Pick C disease proteins. Our approach is generally useful for identifying fluorescent probes in the presence of high cellular autofluorescence

Instantons for Vacuum Decay at Finite Temperature in the Thin Wall Limit

In $N+1$ dimensions, false vacuum decay at zero temperature is dominated by the $O(N+1)$ symmetric instanton, a sphere of radius $R_0$, whereas at temperatures $T>>R_0^{-1}$, the decay is dominated by a `cylindrical' (static) $O(N)$ symmetric instanton. We study the transition between these two regimes in the thin wall approximation. Taking an $O(N)$ symmetric ansatz for the instantons, we show that for $N=2$ and $N=3$ new periodic solutions exist in a finite temperature range in the neighborhood of $T\sim R_0^{-1}$. However, these solutions have higher action than the spherical or the cylindrical one. This suggests that there is a sudden change (a first order transition) in the derivative of the nucleation rate at a certain temperature $T_*$, when the static instanton starts dominating. For $N=1$, on the other hand, the new solutions are dominant and they smoothly interpolate between the zero temperature instanton and the high temperature one, so the transition is of second order. The determinantal prefactors corresponding to the `cylindrical' instantons are discussed, and it is pointed out that the entropic contributions from massless excitations corresponding to deformations of the domain wall give rise to an exponential enhancement of the nucleation rate for $T>>R_0^{-1}$.Comment: 24 pages, 7 figures available upon request, DAMTP-R-94/

Quintessential Maldacena-Maoz Cosmologies

Maldacena and Maoz have proposed a new approach to holographic cosmology based on Euclidean manifolds with disconnected boundaries. This approach appears, however, to be in conflict with the known geometric results [the Witten-Yau theorem and its extensions] on spaces with boundaries of non-negative scalar curvature. We show precisely how the Maldacena-Maoz approach evades these theorems. We also exhibit Maldacena-Maoz cosmologies with [cosmologically] more natural matter content, namely quintessence instead of Yang-Mills fields, thereby demonstrating that these cosmologies do not depend on a special choice of matter to split the Euclidean boundary. We conclude that if our Universe is fundamentally anti-de Sitter-like [with the current acceleration being only temporary], then this may force us to confront the holography of spaces with a connected bulk but a disconnected boundary.Comment: Much improved exposition, exponent in Cai-Galloway theorem fixed, axionic interpretation of scalar explained, JHEP version. 33 pages, 3 eps figure

A system for accurate and automated injection of hyperpolarized substrate with minimal dead time and scalable volumes over a large range

Over recent years hyperpolarization by dissolution dynamic nuclear polarization has become an established technique for studying metabolism in vivo in animal models. Temporal signal plots obtained from the injected metabolite and daughter products, e.g. pyruvate and lactate, can be fitted to compartmental models to estimate kinetic rate constants. Modeling and physiological parameter estimation can be made more robust by consistent and reproducible injections through automation. An injection system previously developed by us was limited in the injectable volume to between 0.6 and 2.4 ml and injection was delayed due to a required syringe filling step. An improved MR-compatible injector system has been developed that measures the pH of injected substrate, uses flow control to reduce dead volume within the injection cannula and can be operated over a larger volume range. The delay time to injection has been minimized by removing the syringe filling step by use of a peristaltic pump. For 100 ll to 10.000 ml, the volume range typically used for mice to rabbits, the average delivered volume was 97.8% of the demand volume. The standard deviation of delivered volumes was 7 ll for 100 ll and 20 ll for 10.000 ml demand volumes (mean S.D. was 9 ul in this range). In three repeat injections through a fixed 0.96 mm O.D. tube the coefficient of variation for the area under the curve was 2%. For in vivo injections of hyperpolarized pyruvate in tumor-bearing rats, signal was first detected in the input femoral vein cannula at 3–4 s post-injection trigger signal and at 9–12 s in tumor tissue. The pH of the injected pyruvate was 7.1 ± 0.3 (mean ± S.D., n = 10). For small injection volumes, e.g. less than 100 ll, the internal diameter of the tubing contained within the peristaltic pump could be reduced to improve accuracy. Larger injection volumes are limited only by the size of the receiving vessel connected to the pump

Prospects for dedicated energy crop production and attitudes towards agricultural straw use: the case of livestock farmers

Second generation biofuels utilising agricultural by-products (e.g. straw), or dedicated energy crops (DECs) produced on ‘marginal’ land, have been called for. A structured telephone survey of 263 livestock farmers, predominantly located in the west or ‘marginal’ upland areas of England captured data on attitudes towards straw use and DECs. Combined with farm physical and business data, the survey results show that 7.2% and 6.3% of farmers would respectively consider growing SRC and miscanthus, producing respective maximum potential English crop areas of 54,603 ha and 43,859 ha. If higher market prices for straw occurred, most livestock farmers would continue to buy straw. Reasons for not being willing to consider growing DECs include concerns over land quality, committing land for a long time period, lack of appropriate machinery, profitability, and time to financial return; a range of moral, land quality, production conflict and lack of crop knowledge factors were also cited. Results demonstrate limited potential for the production of DECs on livestock farms in England. Changes in policy support to address farmer concerns with respect to DECs will be required to incentivise farmers to increase energy crop production. Policy support for DEC production must be cognisant of farm-level economic, tenancy and personal objectives

Increased dietary protein in the second trimester of gestation increases live weight gain and carcass composition in weaner calves to 6 months of age

Genetically similar nulliparous Polled Hereford heifers from a closed pedigree herd were used to evaluate the effects of dietary protein during the first and second trimester of gestation upon fetal, placental and postnatal growth. Heifers were randomly allocated into two groups at 35d post AI (35dpc) to a single bull and fed High (15.7%CP) or Low (5.9%CP) protein in the first trimester (T1). At 90dpc, half of each nutritional treatment group changed to a High or Low protein diet for the second trimester until 180dpc (T2). High protein intake in the second trimester increased birthweight in females (P = 0.05) but there was no effect of treatment upon birthweight when taken over both sexes. Biparietal diameter was significantly increased by high protein in the second trimester with the effect being greater in the female (P = 0.02) but also significant overall (P = 0.05). Placental weight was positively correlated with birth weight, fibroblast volume, and relative blood vessel volume (P < 0.05). Placental fibroblast density was increased and trophoblast volume decreased in the high protein first trimester treatment group (P <0.05). There was a trend for placental weight to be increased by high protein in the second trimester (P = 0.06). Calves from heifers fed the high protein treatment in the second trimester weighed significantly more on all occasions preweaning (at one month (P = 0.0004), 2 mths (P = 0.006), 3 mths (P = 0.002), 4 mths (P = 0.01), 5 mths (P = 41 0.03), 6 mths (P = 0.001)), and grew at a faster rate over the 6 month period. By 6 mths of age the calves from heifers fed high nutrition in the second trimester weighed 33kg heavier than those fed the low diet in the second trimester. These results suggest that dietary protein in early pregnancy alters the development of the bovine placenta and calf growth to weaning

The Physics of Star Cluster Formation and Evolution

© 2020 Springer-Verlag. The final publication is available at Springer via https://doi.org/10.1007/s11214-020-00689-4.Star clusters form in dense, hierarchically collapsing gas clouds. Bulk kinetic energy is transformed to turbulence with stars forming from cores fed by filaments. In the most compact regions, stellar feedback is least effective in removing the gas and stars may form very efficiently. These are also the regions where, in high-mass clusters, ejecta from some kind of high-mass stars are effectively captured during the formation phase of some of the low mass stars and effectively channeled into the latter to form multiple populations. Star formation epochs in star clusters are generally set by gas flows that determine the abundance of gas in the cluster. We argue that there is likely only one star formation epoch after which clusters remain essentially clear of gas by cluster winds. Collisional dynamics is important in this phase leading to core collapse, expansion and eventual dispersion of every cluster. We review recent developments in the field with a focus on theoretical work.Peer reviewe