PAM-2 decreases neuropathic pain in mice and modulates chemokine/cytokine production in human microglial cells

Background: People worldwide suffer from neuropathic pain, and indicated medications are often either not effective or induce tolerance and abuse. Therefore, there is an urgent need to identify additional therapeutic options to treat this form of pain. Nicotinic acetylcholine receptors (nAChRs), particularly a7-nAChRs, have been implicated in pain signaling. Therefore, this study was designed to investigate the extent to which the selective positive allosteric modulator (PAM) of a7 AChRs, PAM-2, modulates neuropathic pain. The working hypothesis, that PAM-2 inhibits inflammatory signaling and neuropathic pain, was tested using animal and cellular models.Methods: The anti-neuropathic pain activity of PAM-2 was assessed in two independent murine models of neuropathic pain. Briefly, neuropathic pain was induced in adult, male CD-1 mice (n=10/condition) via i.p. administration of either streptozotocin (STZ) or oxaliplatin (OXA). After 14 days, when neuropathic pain was present, mice were administrated with PAM-2 (1.0 or 3.0 mg/kg, p.o.) or vehicle. The pain threshold was subsequently determined by the cold plate test before and 15, 30, 45, and 60 min after treatment. In addition, C20 human microglial cells were exposed to interleukin (IL)-1B (20 ng/ml) or vehicle alone, and in combination with nicotine (3 uM), PAM-2 (1-100 uM), or nicotine + PAM-2 for 24 h. After 24 h, cytokine/chemokine levels in the culture media were measured by ELISA.Results: A single dose of PAM-2 (3.0 mg/kg) decreased both STZ- and OXA-induced neuropathic pain in mice. Repeated treatment with an inactive dose (1.0 mg/kg) of PAM-2 showed anti-pain activity in OXA-treated mice after 14, but not 7, days of treatment. Additionally, methyllycaconitine blocked the anti-pain effects elicited by PAM-2, supporting the view that a7 AChRs are instrumental in the anti-pain actions of PAM-2. Cellular experiments revealed that nicotine minimally inhibited IL-1B-induced IL-6 and interferon-gamma-induced chemotactic protein 10 expression in C20 human microglial cells, and that this inhibition was potentiated by PAM-2 (100 uM). However, we cannot rule out the possibility that PAM- 2 was cytotoxic in this cell culture model.Conclusions: These findings indicate that a7 AChRs are involved in neuropathic pain signaling and that a7-PAMs may potentially be used therapeutically. The extent to which these protective effects involve reduced neuroinflammation remains to be determined

Direct in-gel fluorescence detection and cellular imaging of O-GlcNAc-modified proteins

We report an advanced chemoenzymatic strategy for the direct fluorescence detection, proteomic analysis, and cellular imaging of O-GlcNAc-modified proteins. O-GlcNAc residues are selectively labeled with fluorescent or biotin tags using an engineered galactosyltransferase enzyme and [3 + 2] azide−alkyne cycloaddition chemistry. We demonstrate that this approach can be used for direct in-gel detection and mass spectrometric identification of O-GlcNAc proteins, identifying 146 novel glycoproteins from the mammalian brain. Furthermore, we show that the method can be exploited to quantify dynamic changes in cellular O-GlcNAc levels and to image O-GlcNAc-glycosylated proteins within cells. As such, this strategy enables studies of O-GlcNAc glycosylation that were previously inaccessible and provides a new tool for uncovering the physiological functions of O-GlcNAc

Characterization and Correlation of Particle-Level Interactions to the Macroscopic Rheology of Powders, Granular Slurries, and Colloidal Suspensions

This project had two primary objectives. The first was to understand the physical properties and behavior of select Hanford tank sludges under conditions that might exist during retrieval, treatment, packaging, and transportation for disposal at the Waste Isolation Pilot Plant (WIPP). The second objective was to develop a fundamental understanding of these sludge suspensions by correlating the macroscopic properties with particle interactions occurring at the colloidal scale. The specific tank wastes considered herein are contained in thirteen Hanford tanks including three double-shell tanks (DSTs) (AW-103, AW-105, and SY-102) and ten single-shell tanks (SSTs) (B-201 through B-204, T-201 through T-204, T-110, and T-111). At the outset of the project, these tanks were designated as potentially containing transuranic (TRU) process wastes that would be treated and disposed of in a manner different from the majority of the tank wastes

Vitamin D3 supplementation does not enhance the effects of resistance training in older adults

Background: Lifestyle therapy with resistance training is a potent measure to counteract age-related loss in muscle strength and mass. Unfortunately, many individuals fail to respond in the expected manner. This phenomenon is particularly common among older adults and those with chronic diseases (e.g. chronic obstructive pulmonary disease, COPD) and may involve endocrine variables such as vitamin D. At present, the effects of vitamin D supplementation on responses to resistance training remain largely unexplored. Methods: Ninety-five male and female participants (healthy, n = 71; COPD, n = 24; age 68 ± 5 years) were randomly assigned to receive either vitamin D3 or placebo supplementation for 28 weeks in a double-blinded manner (latitude 61°N, September-May). Seventy-eight participants completed the RCT, which was initiated by 12 weeks of supplementation-only (two weeks with 10 000 IU/day, followed by 2000 IU/day), followed by 13 weeks of combined supplementation (2000 IU/day) and supervised whole-body resistance training (twice weekly), interspersed with testing and measurements. Outcome measures included multiple assessments of muscle strength (nvariables = 7), endurance performance (n = 6), and muscle mass (n = 3, legs, primary), as well as muscle quality (legs), muscle biology (m. vastus lateralis; muscle fibre characteristics, transcriptome), and health-related variables (e.g. visceral fat mass and blood lipid profile). For main outcome domains such as muscle strength and muscle mass, weighted combined factors were calculated from the range of singular assessments. Results: Overall, 13 weeks of resistance training increased muscle strength (13% ± 8%), muscle mass (9% ± 8%), and endurance performance (one-legged, 23% ± 15%; whole-body, 8% ± 7%), assessed as weighted combined factors, and were associated with changes in health variables (e.g. visceral fat, -6% ± 21%; [LDL]serum , -4% ± 14%) and muscle tissue characteristics such as fibre type proportions (e.g. IIX, -3% points), myonuclei per fibre (30% ± 65%), total RNA/rRNA abundances (15%/6-19%), and transcriptome profiles (e.g. 312 differentially expressed genes). Vitamin D3 supplementation did not affect training-associated changes for any of the main outcome domains, despite robust increases in [25(OH)D]serum (∆49% vs. placebo). No conditional effects were observed for COPD vs. healthy or pre-RCT [25(OH)D]serum . In secondary analyses, vitamin D3 affected expression of gene sets involved in vascular functions in muscle tissue and strength gains in participants with high fat mass, which advocates further study. Conclusions: Vitamin D3 supplementation did not affect muscular responses to resistance training in older adults with or without COPD. Keywords: Cholecalciferol; Muscle plasticity; Strength training. © 2021 The Authors. Journal of Cachexia, Sarcopenia and Muscle published by John Wiley & Sons Ltd on behalf of Society on Sarcopenia, Cachexia and Wasting Disorders.publishedVersio

Integrating Heterogeneous Odor Response Data into a Common Response Model: A DoOR to the Complete Olfactome

We have developed a new computational framework for merging odor response data sets from heterogeneous studies, creating a consensus metadatabase, the database of odor responses (DoOR). As a result, we obtained a functional atlas of all available odor responses in Drosophila melanogaster. Both the program and the data set are freely accessible and downloadable on the Internet (http://neuro.uni-konstanz.de/DoOR). The procedure can be adapted to other species, thus creating a family of “olfactomes” in the near future. Drosophila melanogaster was chosen because of all species this one is closest to having the complete olfactome characterized, with the highest number of deorphanized receptors available. The database guarantees long-term stability (by offering time-stamped, downloadable versions), up-to-date accuracy (by including new data sets as soon as they are published), and portability (for other species). We hope that this comprehensive repository of odor response profiles will be useful to the olfactory community and to computational neuroscientists alike

Variation in styles of rifting in the Gulf of California

Author Posting. © Nature Publishing Group, 2007. This is the author's version of the work. It is posted here by permission of Nature Publishing Group for personal use, not for redistribution. The definitive version was published in Nature 448 (2007): 466-469, doi:10.1038/nature06035.The rifting of continental lithosphere is a fundamental solid-earth process that leads to the formation of rifted continental margins and ocean basins. Understanding of this process comes from observations of the geometry of rifted margins and the magmatism resulting from rifting, which inform us about the strength of the lithosphere, the state of the underlying mantle, and the transition from rifting to seafloor spreading. Here we describe results from the PESCADOR seismic experiment in the southern Gulf of California and present the first crustal-scale images across conjugate margins of multiple segments within a single rift that has reached the stage of oceanic spreading. A surprisingly large variation in rifting style and magmatism is observed between these segments, from wide rifting with minor syn-rift magmatism to narrow rifting in magmatically robust segments. These differences encompass much of the variation observed across nearly all other non-end-member continental margins. The characteristics of magmatic endmember margins are typically explained in terms of mantle temperature. Our explanations for the variation in the Gulf of California, in contrast, invoke mantle depletion to account for wide, magma-poor rifting and mantle fertility and possibly the influence of sediments to account for robust rift and post-rift magmatism in the Gulf of California. These factors may vary laterally over small distances in regions that have transitioned from convergence to extension, as is the case for the Gulf of California and many other rifts.This work was funded by a grant from the U.S. NSF-MARGINS program

Signal Processing Research Program

Contains table of contents for Part III, table of contents for Section 1, an introduction and reports on fourteen research projects.Charles S. Draper Laboratory Contract DL-H-404158U.S. Navy - Office of Naval Research Grant N00014-89-J-1489National Science Foundation Grant MIP 87-14969Battelle LaboratoriesTel-Aviv University, Department of Electronic SystemsU.S. Army Research Office Contract DAAL03-86-D-0001The Federative Republic of Brazil ScholarshipSanders Associates, Inc.Bell Northern Research, Ltd.Amoco Foundation FellowshipGeneral Electric FellowshipNational Science Foundation FellowshipU.S. Air Force - Office of Scientific Research FellowshipU.S. Navy - Office of Naval Research Grant N00014-85-K-0272Natural Science and Engineering Research Council of Canada - Science and Technology Scholarshi

Serum neurofilament dynamics predicts neurodegeneration and clinical progression in presymptomatic Alzheimer's disease

Neurofilament light chain (NfL) is a promising fluid biomarker of disease progression for various cerebral proteopathies. Here we leverage the unique characteristics of the Dominantly Inherited Alzheimer Network and ultrasensitive immunoassay technology to demonstrate that NfL levels in the cerebrospinal fluid (n = 187) and serum (n = 405) are correlated with one another and are elevated at the presymptomatic stages of familial Alzheimer's disease. Longitudinal, within-person analysis of serum NfL dynamics (n = 196) confirmed this elevation and further revealed that the rate of change of serum NfL could discriminate mutation carriers from non-mutation carriers almost a decade earlier than cross-sectional absolute NfL levels (that is, 16.2 versus 6.8 years before the estimated symptom onset). Serum NfL rate of change peaked in participants converting from the presymptomatic to the symptomatic stage and was associated with cortical thinning assessed by magnetic resonance imaging, but less so with amyloid-β deposition or glucose metabolism (assessed by positron emission tomography). Serum NfL was predictive for both the rate of cortical thinning and cognitive changes assessed by the Mini-Mental State Examination and Logical Memory test. Thus, NfL dynamics in serum predict disease progression and brain neurodegeneration at the early presymptomatic stages of familial Alzheimer's disease, which supports its potential utility as a clinically useful biomarker

Digital Signal Processing Research Program

Contains table of contents for Part III, table of contents for Section 1, an introduction and reports on seventeen research projects.U.S. Navy - Office of Naval Research Contract N00014-90-J-1544Charles S. Draper Laboratory Contract DL-H-404158Rockwell Corporation Doctoral FellowshipU.S. Navy - Office of Naval Research Grant N00014-89-J-1489U.S. Navy - Office of Naval Research Grant N00014-90-J-1109The Federative Republic of Brazil ScholarshipLockheed Sanders, Inc.National Science Foundation Grant MIP 87-14969AT&T Bell Laboratories Doctoral ProgramBell Northern Research Ltd.Defense Advanced Research Projects Agency Contract N00014-87-K-0825IBM CorporationSloan FoundationU.S. Air Force - Office of Scientific Research FellowshipU.S. Air Force - Office of Scientific Research Grant AFOSR-91-0034National Science Foundation Graduate FellowshipCanada, Natural Science and Engineering Research Council ScholarshipU.S. Air Force - Office of Scientific Research Grant AFOSR-91-0034Texas Instruments, Inc

Neutralization Serotyping of BK Polyomavirus Infection in Kidney Transplant Recipients

BK polyomavirus (BKV or BKPyV) associated nephropathy affects up to 10% of kidney transplant recipients (KTRs). BKV isolates are categorized into four genotypes. It is currently unclear whether the four genotypes are also serotypes. To address this issue, we developed high-throughput serological assays based on antibody-mediated neutralization of BKV genotype I and IV reporter vectors (pseudoviruses). Neutralization-based testing of sera from mice immunized with BKV-I or BKV-IV virus-like particles (VLPs) or sera from naturally infected human subjects revealed that BKV-I specific serum antibodies are poorly neutralizing against BKV-IV and vice versa. The fact that BKV-I and BKV-IV are distinct serotypes was less evident in traditional VLP-based ELISAs. BKV-I and BKV-IV neutralization assays were used to examine BKV type-specific neutralizing antibody responses in KTRs at various time points after transplantation. At study entry, sera from 5% and 49% of KTRs showed no detectable neutralizing activity for BKV-I or BKV-IV neutralization, respectively. By one year after transplantation, all KTRs were neutralization seropositive for BKV-I, and 43% of the initially BKV-IV seronegative subjects showed evidence of acute seroconversion for BKV-IV neutralization. The results suggest a model in which BKV-IV-specific seroconversion reflects a de novo BKV-IV infection in KTRs who initially lack protective antibody responses capable of neutralizing genotype IV BKVs. If this model is correct, it suggests that pre-vaccinating prospective KTRs with a multivalent VLP-based vaccine against all BKV serotypes, or administration of BKV-neutralizing antibodies, might offer protection against graft loss or dysfunction due to BKV associated nephropathy