The role of the AFD neuron in C. elegans thermotaxis analyzed using femtosecond laser ablation

BACKGROUND: Caenorhabditis elegans actively crawls down thermal gradients until it reaches the temperature of its prior cultivation, exhibiting what is called cryophilic movement. Implicit in the worm's performance of cryophilic movement is the ability to detect thermal gradients, and implicit in regulating the performance of cryophilic movement is the ability to compare the current temperature of its surroundings with a stored memory of its cultivation temperature. Several lines of evidence link the AFD sensory neuron to thermotactic behavior, but its precise role is unclear. A current model contends that AFD is part of a thermophilic mechanism for biasing the worm's movement up gradients that counterbalances the cryophilic mechanism for biasing its movement down gradients. RESULTS: We used tightly-focused femtosecond laser pulses to dissect the AFD neuronal cell bodies and the AFD sensory dendrites in C. elegans to investigate their contribution to cryophilic movement. We establish that femtosecond laser ablation can exhibit submicrometer precision, severing individual sensory dendrites without causing collateral damage. We show that severing the dendrites of sensory neurons in young adult worms permanently abolishes their sensory contribution without functional regeneration. We show that the AFD neuron regulates a mechanism for generating cryophilic bias, but we find no evidence that AFD laser surgery reduces a putative ability to generate thermophilic bias. In addition, although disruption of the AIY interneuron causes worms to exhibit cryophilic bias at all temperatures, we find no evidence that laser killing the AIZ interneuron causes thermophilic bias at any temperature. CONCLUSION: We conclude that laser surgical analysis of the neural circuit for thermotaxis does not support a model in which AFD opposes cryophilic bias by generating thermophilic bias. Our data supports a model in which the AFD neuron gates a mechanism for generating cryophilic bias

The Effect of Body Size on Countermovement Jump Kinetics in Children aged 7 to 11 years

The purpose this study was to examine the effect of body size oncountermovement jump (CMJ)kinetics in children.Participants(n = 160) aged 7-11 years, divided equally by sex and into primary school year groups(years 3, 4, 5 and 6), each performedone CMJ on aforce platform. The variables bodyweight(BW), peak force (Fmax), in-jump minimum force (IMF), in-jump vertical force range (IFR) and basic rate of force development (BRFD)wereattained from the force-time history and then subsequently scaled to account for body size. A significant age, sex and interaction effect werefound for theabsolutevariables BW, IMF, Fmaxand IFR (P 0.05). No significant age or sex differences were observed for normalised or allometrically scaled values(P > 0.05). The results indicate thatgirls and boys can be grouped together but that body size must be accounted for to enable accurate conclusions to be drawn independent of growth.Bodysizesignificantlyeffects the representation of CMJ kinetic results and therefore, future studies should report both absolute and scaled values.Future research should developan age-appropriate criterion method for children in order to determine processed CMJ variables to further investigate neuromuscular performance of children

Altered Cytokine Production in Mice Lacking P2X 7 Receptors

The P2X(7) receptor (P2X(7)R) is an ATP-gated ion channel expressed by monocytes and macrophages. To directly address the role of this receptor in interleukin (IL)-1 beta post-translational processing, we have generated a P2X(7)R-deficient mouse line. P2X(7)R(-/-) macrophages respond to lipopolysaccharide and produce levels of cyclooxygenase-2 and pro-IL-1 beta comparable with those generated by wild-type cells. In response to ATP, however, pro-IL-1 beta produced by the P2X(7)R(-/-) cells is not externalized or activated by caspase-1. Nigericin, an alternate secretion stimulus, promotes release of 17-kDa IL-1 beta from P2X(7)R(-/-) macrophages. In response to in vivo lipopolysaccharide injection, both wild-type and P2X(7)R(-/-) animals display increases in peritoneal lavage IL-6 levels but no detectable IL-1. Subsequent ATP injection to wild-type animals promotes an increase in IL-1, which in turn leads to additional IL-6 production; similar increases did not occur in ATP-treated, LPS-primed P2X(7)R(-/-) animals. Absence of the P2X(7)R thus leads to an inability of peritoneal macrophages to release IL-1 in response to ATP. As a result of the IL-1 deficiency, in vivo cytokine signaling cascades are impaired in P2X(7)R-deficient animals. Together these results demonstrate that P2X(7)R activation can provide a signal that leads to maturation and release of IL-1 beta and initiation of a cytokine cascade

Identification and characterization of a novel class of interleukin-1 post-translational processing inhibitors

ABSTRACT Lipopolysaccharide (LPS)-activated monocytes and macrophages produce large quantities of pro-interleukin (IL)-1␤ but externalize little mature cytokine. Efficient post-translational processing of the procytokine occurs in vitro when these cells encounter a secretion stimulus such as ATP, cytolytic T cells, or hypotonic stress. Each of these stimuli promotes rapid conversion of 31-kDa pro-IL-1␤ to its mature 17-kDa species and release of the 17-kDa cytokine. In this study, two novel pharmacological agents, CP-424,174 and CP-412,245, are identified as potent inhibitors of stimulus-coupled IL-1␤ post-translational processing. These agents, both diarylsulfonylureas, block formation of mature IL-1␤ without increasing the amount of procytokine that is released extracellularly, and they inhibit independently of the secretion stimulus used. Conditioned medium derived from LPS-activated/ATP-treated human monocytes maintained in the absence and presence of CP-424,174 contained comparable quantities of IL-6, tumor necrosis factor-␣ (TNF␣), and IL-1RA, but 30-fold less IL-1␤ was generated in the test agent's presence. As a result of this decrease, monocyte conditioned medium prepared in the presence of CP-424,174 demonstrated a greatly diminished capacity to promote an IL-1-dependent response (induction of serum amyloid A synthesis by Hep3B cells). Oral administration of CP-424,174 to mice resulted in inhibition of IL-1 in the absence of an effect on IL-6 and TNF␣. These novel agents, therefore, act as selective cytokine release inhibitors and define a new therapeutic approach for controlling IL-1 production in inflammatory diseases

Galactic-scale absorption outflow in the low-luminosity quasar IRAS F04250-5718: Hubble space telescope/cosmic origins spectrograph observations

We present absorption line analysis of the outflow in the quasar IRAS F04250?5718. Far-ultraviolet data from the Cosmic Origins Spectrograph on board the Hubble Space Telescope reveal intrinsic narrow absorption lines from high ionization ions (e.g., C iv, N v, and O vi) as well as low ionization ions (e.g., Cii and Si iii). We identify three kinematic components with central velocities ranging from ??50 to ??230 km s?1. Velocity-dependent, nonblack saturation is evident from the line profiles of the high ionization ions. From the non-detection of absorption from a metastable level of C ii, we are able to determine that the electron number density in the main component of the outflow is 30 cm?3. Photoionization analysis yields an ionization parameter log UH ? ?1.6 ± 0.2, which accounts for changes in the metallicity of the outflow and the shape of the incident spectrum. We also consider solutions with two ionization parameters. If the ionization structure of the outflow is due to photoionization by the active galactic nucleus, we determine that the distance to this component from the central source is 3 kpc. Due to the large distance determined for the main kinematic component, we discuss the possibility that this outflow is part of a galactic wind.We acknowledge support from NASA STScI grants GO 11686 and GO 12022 as well as NSF grant AST 0837880. We thank Pat Hall for insightful suggestions and discussions. J.I.G.-S. and C.B. acknowledge financial support from the Spanish Ministerio de Ciencia e Innovacion under project AYA2008-06311-C02-02

Functional and epigenetic phenotypes of humans and mice with DNMT3A Overgrowth Syndrome

Germline mutations in the DNMT3A gene can cause an overgrowth syndrome associated with behavioural and hematopoietic phenotypes. Here the authors describe a mouse model of this syndrome that recapitulates many of these features, including conserved alterations in DNA methylation in the blood cells of both species

The Core Apoptotic Executioner Proteins CED-3 and CED-4 Promote Initiation of Neuronal Regeneration in Caenorhabditis elegans

Laser severing of individual axons in the nematode Caenorhabditis elegans revealed that the apoptotic executioner caspase CED-3 and its regulator CED-4/Apaf-1 play an unexpected beneficial role in promoting axonal regeneration

P2 purinergic receptor modulation of cytokine production

Cytokines serve important functions in controlling host immunity. Cells involved in the synthesis of these polypeptide mediators have evolved highly regulated processes to ensure that production is carefully balanced. In inflammatory and immune disorders, however, mis-regulation of the production and/or activity of cytokines is recognized as a major contributor to the disease process, and therapeutics that target individual cytokines are providing very effective treatment options in the clinic. Leukocytes are the principle producers of a number of key cytokines, and these cells also express numerous members of the purinergic P2 receptor family. Studies in several cellular systems have provided evidence that P2 receptor modulation can affect cytokine production, and mechanistic features of this regulation have emerged. This review highlights three separate examples corresponding to (1) P2Y6 receptor mediated impact on interleukin (IL)-8 production, (2) P2Y11 receptor-mediated affects on IL-12/23 output, and (3) P2X7 receptor mediated IL-1β posttranslational processing. These examples demonstrate important roles of purinergic receptors in the modulation of cytokine production. Extension of these cellular observations to in vivo situations may lead to new therapeutic strategies for treating cytokine-mediated diseases