The endogenous phospholipid, lysophosphatidic acid (LPA), is known to mediate an array of physiologic processes, including proliferation and cell migration. In addition, LPA signaling is implicated in the pathophysiology of fibrotic disease and evokes a pro‐inflammatory response. The various effects of LPA are mediated by interaction with one of six cognate receptors, termed LPA 1–6 . In particular, many of the pro‐fibrotic and pro‐inflammatory effects of LPA are attributed to interaction with the LPA 1 receptor. LPA 1 receptor stimulation promotes signaling through G q , G i/o , G 12/13 and arrestin. Notably, while LPA species 18:1 is equipotent at evoking each of these downstream signals, we identified antagonists that exhibit biased inhibition of the various pathways. To better understand the molecular pharmacology of LPA signaling in renal fibrosis and inflammation, we studied the signaling pathways activated by 18:1 LPA in various primary human renal cell types, including proximal tubule epithelial cells and renal fibroblasts. These studies enabled mapping of the complex LPA signaling network that drives key pro‐fibrotic and pro‐inflammatory endpoints. Finally, we will present efficacy data comparing full antagonists against antagonists with various biased profiles in the mouse UUO model of renal fibrosis. In total, these data demonstrate the complexity of the LPA‐LPA 1 signaling events that drive renal fibrosis, and highlight the requisite antagonist profiles to reverse renal fibrosis endpoints in vivo. Disclosures: All authors are employees of AbbVie. The design, study conduct, and financial support for this research were provided by AbbVie. AbbVie participated in the interpretation of data, review, and approval of the publication. Support or Funding Information All research was funded by AbbVie Inc.
N-, T- and P/Q-type voltage-gated Ca 2+ channels are critical for regulating neurotransmitter release and cellular excitability and have been implicated in mediating pathological nociception. A-1264087 is a novel state-dependent blocker of N-, T- and P/Q-type channels. In the present studies, A-1264087 blocked (IC 50 = 1.6 μM) rat dorsal root ganglia N-type Ca 2+ in a state-dependent fashion. A-1264087 (1, 3 and 10 mg/kg po) dose-dependently reduced mechanical allodynia in rats with a spinal nerve ligation (SNL) injury. A-1264087 (4 mg/kg iv) inhibited both spontaneous and mechanically evoked activity of spinal wide dynamic range (WDR) neurons in SNL rats but had no effect in uninjured rats. The inhibitory effect on WDR neurons remained in spinally transected SNL rats. Injection of A-1264087 (10 nmol/0.5 μl) into the spinal cord reduced both spontaneous and evoked WDR activity in SNL rats. Application of A-1264087 (300 nmol/20 μl) into the receptive field on the hindpaw attenuated evoked but not spontaneous firing of WDR neurons. Using electrical stimulation, A-1264087 (4 mg/kg iv) inhibited Aδ- and C-fiber evoked responses and after-discharge of WDR neurons in SNL rats. These effects by A-1264087 were not present in uninjured rats. A-1264087 moderately attenuated WDR neuron windup in both uninjured and SNL rats. In summary, these results indicate that A-1264087 selectively inhibited spinal nociceptive transmission in sensitized states through both peripheral and central mechanisms.
In this paper we report on the modeling and characterization of transmission windows for in-situ interferometric measurements of cryogenically cooled mirrors. Specifically, we present a model of the temperature distribution and strain in the transmission window, and the corresponding spatial dependence of the window's index of refraction. We also present experimental results which characterize the effect of the windows on interferometric measurement.
An ongoing collaboration among four US Department of Energy (DOE) National Laboratories has demonstrated key technology prototypes and software modeling tools required for new high-coherent flux beamline optical systems. New free electron laser (FEL) and diffraction-limited storage ring (DLSR) light sources demand wavefront preservation from source to sample to achieve and maintain optimal performance. Fine wavefront control was achieved using a novel, roomtemperature cooled mirror system called REAL (resistive element adjustable length) that combines cooling with applied, spatially variable auxiliary heating. Single-grating shearing interferometry (also called Talbot interferometry) and Hartmann wavefront sensors were developed and used for optical characterization and alignment on several beamlines, across a range of photon energies. Demonstrations of non-invasive hard x-ray wavefront sensing were performed using a thin diamond single-crystal as a beamsplitter.
Patients with primary ciliary dyskinesia (PCD) suffer from a variety of disorders that include chronic sinusitis, otitis media, infertility, situs inversus, and hydrocephalus. We have shown that two mouse models of PCD, nm1054 and bgh, have hydrocephalus characterized by dilatation of the lateral ventricles. These models also have male infertility and sinus abnormalities that result from a decrease in respiratory ciliary beat frequency. We therefore hypothesize that the hydrocephalus results from dysfunction of ependymal cilia that prevents proper cerebrospinal fluid (CSF) flow. To investigate CSF flow in vivo, we injected India ink into the lateral ventricles of live mice and histologically demonstrated that ink migrated through the ventricular system of wild type mice but failed to progress past the cerebral aqueduct of either mutant. To validate that this defect was directly due to ciliary dysfunction, we used an ex vivo method to measure ink flow rate over the ependymal cells on dissected ventricular walls. This method demonstrated a decreased flow rate and altered directional flow in the mutant brains, confirming that ependymal ciliary function is required for proper CSF flow. Grant Funding Source : Supported by the NIH NIGMS (1P20GM103620‐01A1) and an NSF REU (1262744)
With the nearly full spatial coherence of X-ray free electron lasers comes unprecedented requirements on the precise figure of X-ray mirrors. For example, the Time-resolved AMO Instrument (TMO) requires a bendable Kirkpatrick-Baez (KB) mirror system in order to produce a range of focus spot sizes, while maintaining nm-level figure error across the corresponding range of elliptical shapes. Here we describe the process of TMO KB mirror characterization using optical metrology, the detailed comparison with at-wavelength wavefront sensors during instrument commissioning, and the implementation of an automatic focusing system that ties together the wavefront sensor output with the mirror bender controls.
Primary ciliary dyskinesia (PCD) is a genetically heterogeneous syndrome that results from defects in motile cilia. The ciliary axoneme has a 9 + 2 microtubule structure consisting of nine peripheral doublets surrounding a central pair apparatus (CPA), which plays a critical role in regulating proper ciliary function. We have previously shown that mouse models with mutations in CPA genes CFAP221, CFAP54, and SPEF2 have a PCD phenotype with defects in ciliary motility. In this study, we investigated potential genetic interaction between these CPA genes by generating each combination of double heterozygous and double homozygous mutants. No detectable cilia-related phenotypes were observed in double heterozygotes, but all three double homozygous mutant lines exhibit early mortality and typically develop severe PCD-associated phenotypes of hydrocephalus, mucociliary clearance defects in the upper airway, and abnormal spermatogenesis. Double homozygous cilia are generally intact and display a normal morphology and distribution. Spermiogenesis is aborted in double homozygotes, with an absence of mature flagella on elongating spermatids and epididymal sperm. These findings identify genetic interactions between CPA genes and genetic mechanisms regulating the CPA and motile cilia function.
With the onset of high power XFELs and diffraction limited storage rings, there is a growing demand to maintain sub nanometer mirror figures even under high heat load. This is a difficult issue as the optimum cooling design for an optic is highly dependent on the power footprint on the mirror, which can be highly dynamic. Resistive Element Adjustable Length (REAL) cooling can be utilized to change the cooling parameters during an experiment to adapt for changing beam parameters. A case study of the new soft x-ray monochromator for the LCLS L2SI program is presented that utilizes this new capability to allow the beam to translate across the mirror for different operation modes, greatly simplifying the monochromator mechanics. Metrology of a prototype mirror will also be presented.
Congenital hydrocephalus results from accumulation of cerebrospinal fluid (CSF) and causes severe neurological damage, but the underlying causes are not well understood. Hydrocephalus is associated with primary ciliary dyskinesia (PCD), a pediatric syndrome caused by dysfunction of motile cilia that can also result in sinusitis, male infertility, and situs inversus. We have investigated the causes of hydrocephalus in mouse models of PCD lacking the Pcdp1 and Spef2 proteins. Both models develop gross, lethal hydrocephalus on the C57BL/6J (B6) background but not on 129S6/SvEvTac (129). Histological and immunohistochemical analyses demonstrate that both mutants on B6 have ventricular dilatation, sloughing of ciliated ependyma, disrupted axon myelination, and increased gliosis, indicating significant injury to the cerebral cortex. In contrast, both mutants on both the B6 and 129 backgrounds exhibit perturbed CSF flow despite an absence of substantial morphological defects on 129. These findings indicate that abnormal cilia‐driven flow is not the sole determinant of PCD‐associated hydrocephalus and suggest that there are genetic modifiers that generally influence susceptibility to congenital hydrocephalus. Grant Funding Source : Supported by the NIH (1P20GM103620‐01A1)
