Rictor/TORC2 Regulates Fat Metabolism, Feeding, Growth and Life Span in C. elegans

Rictor is a component of the target of rapamycin complex 2 (TORC2). While TORC2 has been implicated in insulin and other growth factor signaling pathways, the key inputs and outputs of this kinase complex remain unknown. We identified mutations in the Caenorhabditis elegans homolog of rictor in a forward genetic screen for increased body fat. Despite high body fat, rictor mutants are developmentally delayed, small in body size, lay an attenuated brood, and are short-lived, indicating that Rictor plays a critical role in appropriately partitioning calories between long-term energy stores and vital organismal processes. Rictor is also necessary to maintain normal feeding on nutrient-rich food sources. In contrast to wild-type animals, which grow more rapidly on nutrient-rich bacterial strains, rictor mutants display even slower growth, a further reduced body size, decreased energy expenditure, and a dramatically extended life span, apparently through inappropriate, decreased consumption of nutrient-rich food. Rictor acts directly in the intestine to regulate fat mass and whole-animal growth. Further, the high-fat phenotype of rictor mutants is genetically dependent on akt-1, akt-2, and serum and glucocorticoid-induced kinase-1 (sgk-1). Alternatively, the life span, growth, and reproductive phenotypes of rictor mutants are mediated predominantly by sgk-1. These data indicate that Rictor/TORC2 is a nutrient-sensitive complex with outputs to AKT and SGK to modulate the assessment of food quality and signal to fat metabolism, growth, feeding behavior, reproduction, and life span

Biochemical and High Throughput Microscopic Assessment of Fat Mass in Caenorhabditis Elegans

The nematode C. elegans has emerged as an important model for the study of conserved genetic pathways regulating fat metabolism as it relates to human obesity and its associated pathologies. Several previous methodologies developed for the visualization of C. elegans triglyceride-rich fat stores have proven to be erroneous, highlighting cellular compartments other than lipid droplets. Other methods require specialized equipment, are time-consuming, or yield inconsistent results. We introduce a rapid, reproducible, fixative-based Nile red staining method for the accurate and rapid detection of neutral lipid droplets in C. elegans. A short fixation step in 40% isopropanol makes animals completely permeable to Nile red, which is then used to stain animals. Spectral properties of this lipophilic dye allow it to strongly and selectively fluoresce in the yellow-green spectrum only when in a lipid-rich environment, but not in more polar environments. Thus, lipid droplets can be visualized on a fluorescent microscope equipped with simple GFP imaging capability after only a brief Nile red staining step in isopropanol. The speed, affordability, and reproducibility of this protocol make it ideally suited for high throughput screens. We also demonstrate a paired method for the biochemical determination of triglycerides and phospholipids using gas chromatography mass-spectrometry. This more rigorous protocol should be used as confirmation of results obtained from the Nile red microscopic lipid determination. We anticipate that these techniques will become new standards in the field of C. elegans metabolic research

Overview of the ATR power supplies

The AGS to RHIC transfer line (ATR) transports a variety of beams from the Alternating Gradient Synchrotron (AGS) which gets its input from the Booster Synchrotron. In turn, the Booster receives input beams from either a Tandem Van de Graaff (heavy ions) or a Linac (protons). The AGS extracts beam bunches, up to a rate of 30 Hertz, to the ATR which feeds the Relativistic Heavy Ion Collider (RHIC) starting with the sextant test in January of 1997. The ATR is made up of the upgraded U line and the new W, X, and Y lines. A test in 1995 transported beam to the end of the W line. During normal operation, a pulsed switching magnet at the end of the W line will bend the beam into the X line or the Y line so that the two storage rings in RHIC are filled with counter rotating beams. The ATR line is comprised of 80 power supplies (PS`s), 17 of which are upgraded AGS PS`s. The remaining 63 PS`s were newly purchased. These PS`s range from bipolar 600 watt linear type trim magnet PS`s to 1 Megawatt, thyristor, dipole PS`s. Results of the commissioning runs will be presented, as well as descriptions of regulation, filtering, and analog and digital controls

AGS slow extracted beam improvement

The Brookhaven AGS is a strong focusing accelerator which is used to accelerate protons and various heavy ion species to an equivalent proton energy of 29 GeV. Since the late 1960`s it has been serving high energy physics (HEP - proton beam) users of both slow and fast extracted beams. The AGS fixed target program presently uses primary proton and heavy ion beams (HIP) in slowly extracted fashion over spill lengths of 1.5 to 4.0 seconds. Extraction is accomplished by flattoping the main and extraction magnets and exciting a third integer resonance in the AGS. Over the long spill times, control of the subharmonic amplitude components up to a frequency of 1 kilohertz is very crucial. One of the most critical contributions to spill modulation is due to the AGS MMPS. An active filter was developed to reduce these frequencies and it`s operation is described in a previous paper. However there are still frequency components in the 60-720 Hz sub-harmonic ripple range, modulating the spill structure due to extraction power supplies and any remaining structures on the AGS MMPS. A recent scheme is being developed to use the existing tune-trim control horizontal quadrupole magnets and power supply to further reduce these troublesome noise sources. Feedback from an external beam sensor and overcoming the limitations of the quadrupole system by lead/lag compensation techniques will be described

Test fast kicker pulser

In this paper, a test pulser of the Brookhaven AGS Booster extraction fast kicker is described. The pulser is projected for both proton and heavy ion operation. A load of total inductance 2.15 /mu/H is used for the test pulser. The PFN voltage is required to be below 40 kV for operation in air. Rise time of the pulse for proton extraction operation is about 120ns up to 97% of full current (1000A), and, for heavy ion extraction, 160ns up to 98% of full current (1615A). R-C compensation networks are used for pulse front edge sharpening. The flexibility of operation is obtained basically by switching an energy dumping resistor to match or mismatch the PFN impedance. Some comments on stray capacitance and stray inductance effects are included. 3 refs., 10 figs., 2 tabs

Redirection of SKN-1 abates the negative metabolic outcomes of a perceived pathogen infection

Early host responses toward pathogens are essential for defense against infection. In Caenorhabditis elegans, the transcription factor, SKN-1, regulates cellular defenses during xenobiotic intoxication and bacterial infection. However, constitutive activation of SKN-1 results in pleiotropic outcomes, including a redistribution of somatic lipids to the germline, which impairs health and shortens lifespan. Here, we show that exposing C. elegans to Pseudomonas aeruginosa similarly drives the rapid depletion of somatic, but not germline, lipid stores. Modulating the epigenetic landscape refines SKN-1 activity away from innate immunity targets, which alleviates negative metabolic outcomes. Similarly, exposure to oxidative stress redirects SKN-1 activity away from pathogen response genes while restoring somatic lipid distribution. In addition, activating p38/MAPK signaling in the absence of pathogens, is sufficient to drive SKN-1-dependent loss of somatic fat. These data define a SKN-1- and p38-dependent axis for coordinating pathogen responses, lipid homeostasis, and survival and identify transcriptional redirection, rather than inactivation, as a mechanism for counteracting the pleiotropic consequences of aberrant transcriptional activity

Limits on the production of neutral penetrating states in a beam dump

We present limits on the production of neutral penetrating states produced in 28 GeV proton nucleus collisions. We obtain limits for light, heavy and unstable neutral states. For light stable states our limit [sigma]I[sigma]P-69cm4/nucleon2 is more than a factor of 5.5 better than previous limits. Time of flight techniques are used to study heavy states. We have poor sensitivity to short-lived states.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/24349/1/0000616.pd

Clustering gene expression data with a penalized graph-based metric

<p>Abstract</p> <p>Background</p> <p>The search for cluster structure in microarray datasets is a base problem for the so-called "-omic sciences". A difficult problem in clustering is how to handle data with a manifold structure, i.e. data that is not shaped in the form of compact clouds of points, forming arbitrary shapes or paths embedded in a high-dimensional space, as could be the case of some gene expression datasets.</p> <p>Results</p> <p>In this work we introduce the Penalized k-Nearest-Neighbor-Graph (PKNNG) based metric, a new tool for evaluating distances in such cases. The new metric can be used in combination with most clustering algorithms. The PKNNG metric is based on a two-step procedure: first it constructs the k-Nearest-Neighbor-Graph of the dataset of interest using a low k-value and then it adds edges with a highly penalized weight for connecting the subgraphs produced by the first step. We discuss several possible schemes for connecting the different sub-graphs as well as penalization functions. We show clustering results on several public gene expression datasets and simulated artificial problems to evaluate the behavior of the new metric.</p> <p>Conclusions</p> <p>In all cases the PKNNG metric shows promising clustering results. The use of the PKNNG metric can improve the performance of commonly used pairwise-distance based clustering methods, to the level of more advanced algorithms. A great advantage of the new procedure is that researchers do not need to learn a new method, they can simply compute distances with the PKNNG metric and then, for example, use hierarchical clustering to produce an accurate and highly interpretable dendrogram of their high-dimensional data.</p