A Nicotinic Acetylcholine Receptor Agonist Prevents Loss of Retinal Ganglion Cells in a Glaucoma Model

Purpose.: The purpose of this study was to analyze the neuroprotective effect of an α7 nAChR agonist, PNU-282987, using an in vivo model of glaucoma in Long Evans rats. Methods.: One eye in each animal was surgically manipulated to induce glaucoma in control untreated animals and in animals that were treated with intravitreal injections of PNU-282987. To induce glaucoma-like conditions, 0.05 mL of 2 M NaCl was injected into the episcleral veins of right eyes in each rat to create scar tissue and increase intraocular pressure. The left eye in each rat acted as an internal control. One month following NaCl injection, rats were euthanized, retinas were removed, flatmounted, fixed, and nuclei were stained with cresyl violet or RGCs were immunostained with an antibody against Thy 1.1 or against Brn3a. Stained nuclei in the RGC layer and labeled RGCs in NaCl-injected retinas were counted and compared with cell counts from untreated retinas in the same animal. Results.: NaCl injections into the episcleral veins caused a significant loss of cells by an average of 27.35% (±2.12 SEM) in the RGC layer within 1 month after NaCl injection, which corresponded to a significant loss of RGCs. This loss of RGCs was eliminated if 5 μL of 100 μM PNU-282987 was injected into the right eye an hour before NaCl injection. Conclusions.: The results from this study support the hypothesis that the α7 agonist, PNU-282987, has a neuroprotective effect in the rat retina. PNU-282987 may be a viable candidate for future therapeutic treatments of glaucoma

Prevention of Glaucoma-Induced Retinal Ganglion Cell Loss Using Alpha7 nAChR Agonists

In this study, the neuroprotective effect of various nicotinic alpha7 acetylcholine receptor agonists in an in-vivo model of glaucoma using adult Long Evans rats was analyzed. Glaucoma-like conditions were induced in the eyes of Long Evans rats after injection of hypertonic saline into episcleral veins to create scar tissue and increase the animal’s intraocular pressure. This procedure produced significant loss of retinal ganglion cells within one month and was associated with an increase of intraocular pressure. Using this model system, various alpha7 nicotinic acetylcholine receptor (a7 nAChR) agonists were applied at different doses as eye drops to the right eye of adult Long Evans rats while the left eye was left as an internal control. The a7 nAChR agonists used in this study prevented loss of RGCs in a dose dependent manner after the procedure to induce glaucoma-like conditions. PHA-543613 and PNU- 282987 provided the largest degree of RGC survival after inducing glaucomalike conditions, followed by nicotine, SEN 12333, tropisetron, 3-Bromocytisine and DMAB. To provide evidence that neuroprotection of RGCs was mediated through activation of a7 nAChR, in some studies different concentrations of the a7 nAChR antagonist, MLA, was intravitreally injected into experimentally treated eyes before initiation of eye drops and the procedure to induce glaucoma-like conditions. In the presence of MLA, RGC neuroprotection was blocked. Results from these studies suggest that selective a7 nAChR agonists may be used in future therapeutic treatments for glaucoma or other CNS diseases associated with a7 nAChRs

Target identification and validation of the alpha7 nicotinic acetylcholine receptor as a potential therapeutic target in retinal disease

The role of acetylcholine (ACh) in visual processing in the mammalian retina has been the focus of research for many decades. Pioneering work on the localization of ACh discovered that the neurotransmitter is synthesized and stored in a distinct subpopulation of amacrine (starburst) cells. It has been shown that ACh release is regulated to a low resting “tonic” level, much like what is observed at the neuromuscular junction (NMJ). If there were a dysfunction in the tonic release of ACh, might post-synaptic changes render the targets of ACh [i.e., retinal ganglion cells (RGCs)] vulnerable to disease? During my time at Pharmacia & Upjohn (PNU), selective nicotinic ACh receptor (nAChR) agonists (e.g., PNU-282987) were developed as a possible therapy for central nervous system (CNS) diseases. As RGCs are the main targets of neurodegeneration in glaucoma, could the activation of this target provide neuroprotection? In response to this question, experiments to identify alpha7 nAChRs in the retina (i.e., target ID studies) followed by “proof-of-concept” experiments were conducted. Target ID studies included binding studies with retinal homogenates, [125I]-alpha-bungarotoxin (α-BTX) autoradiography, and fluorescently tagged α-BTX binding in retinal slices. Imaging studies of intracellular calcium dynamics in the retinal slice were conducted. Reverse transcription-polymerase chain reaction (RT-PCR) analysis with alpha7 nAChR knockout mice using the “laser-capture microdissection” technique, in situ hybridization studies, and RT-PCR analysis of the human retina were conducted. Collectively, these experiments confirmed the presence of alpha7 nAChRs on specific cells in the retina. “Proof-of-concept” neuroprotection studies demonstrated that PNU-282987 provided significant protection for RGCs. This protection was dose dependent and was blocked with selective antagonists. More recently, evidence for the generation of new RGCs has been reported with PNU-282987 in rodents. Interestingly, the appearance of new RGCs is more pronounced with eye-drop application than with intravitreal injection. One could postulate that this reflects the neurogenic activation of alpha7 receptors on the retinal pigment epithelium (RPE) (eye drops) vs. a neuroprotective effect on RGCs (injections). In conclusion, there does appear to be a cholinergic retinal “tone” associated with RGCs that could be utilized as a neuroprotective therapy. However, a distinct cholinergic neurogenic mechanism also appears to exist in the outer retina that could possibly be exploited to generate new RGCs lost through various disease processes

An Archaeological Inventory of Camp Swift, Bastrop County, Texas

Beginning in November 1996 and continuing until July of 1997, the Adjutant General’s Department of Texas conducted a self-sponsored Phase I cultural resources survey of Camp Swift in Bastrop County, Texas. The project surveyed approximately 5,000 acres of the camp, approximately 1,000 of which had been previously surveyed. A total of 58 new archaeological sites were recorded, of which 26 were prehistoric, 24 were historic, and 8 had both prehistoric and historic components. In addition to these sites, 42 previously identified sites were revisited. In September 2000, the Center for Archaeological Research of the University of Texas at San Antonio completed shovel tests on two sites and acquired GPS data on 28 sites. At this time, a geomorphologist excavated a series of 12 backhoe trenches and—subsequently—a report on the geoarchaeology of Camp Swift was added to this report. An assessment of the 169 sites now known on Camp Swift found one site (41BP138, the Wine Cellar Site) eligible for inclusion in the National Register of Historic Places. A total of 106 sites are considered not eligible for inclusion in the National Register of Historic Places. Fifty-nine sites are considered potentially eligible, and should be tested to determine their eligibility. Three marked cemeteries are protected by state law

Species delimitation in lemurs: multiple genetic loci reveal low levels of species diversity in the genus Cheirogaleus

<p>Abstract</p> <p>Background</p> <p>Species are viewed as the fundamental unit in most subdisciplines of biology. To conservationists this unit represents the currency for global biodiversity assessments. Even though Madagascar belongs to one of the top eight biodiversity hotspots of the world, the taxonomy of its charismatic lemuriform primates is not stable. Within the last 25 years, the number of described lemur species has more than doubled, with many newly described species identified among the nocturnal and small-bodied cheirogaleids. Here, we characterize the diversity of the dwarf lemurs (genus <it>Cheirogaleus</it>) and assess the status of the seven described species, based on phylogenetic and population genetic analysis of mtDNA (<it>cytb </it>+ <it>cox2</it>) and three nuclear markers (<it>adora3</it>, <it>fiba </it>and <it>vWF</it>).</p> <p>Results</p> <p>This study identified three distinct evolutionary lineages within the genus <it>Cheirogaleus</it>. Population genetic cluster analyses revealed a further layer of population divergence with six distinct genotypic clusters.</p> <p>Conclusion</p> <p>Based on the general metapopulation lineage concept and multiple concordant data sets, we identify three exclusive groups of dwarf lemur populations that correspond to three of the seven named species: <it>C. major</it>, <it>C. medius </it>and <it>C. crossleyi</it>. These three species were found to be genealogically exclusive in both mtDNA and nDNA loci and are morphologically distinguishable. The molecular and morphometric data indicate that <it>C. adipicaudatus </it>and <it>C. ravus </it>are synonymous with <it>C. medius </it>and <it>C. major</it>, respectively. <it>Cheirogaleus sibreei </it>falls into the <it>C. medius </it>mtDNA clade, but in morphological analyses the membership is not clearly resolved. We do not have sufficient data to assess the status of <it>C. minusculus</it>. Although additional patterns of population differentiation are evident, there are no clear subdivisions that would warrant additional specific status. We propose that ecological and more geographic data should be collected to confirm these results.</p

Low-Cost Flow Visualization for a Supersonic Ejector

Shadowgraph techniques were applied to the cold flow ejector facility at the Propulsion Research Center at the University of Alabama in Huntsville. The setup for the experiments was relatively simple and was accomplished at very little cost. Series of shadowgraph images were taken of both dual nozzle and single nozzle strut based ejectors operating over a range of chamber pressures. The density gradient patterns in the shadowgraphs were compared to pressure data measured along the top and side walls of the mixing duct. The shadowgraph images showed the presence of barrel shocks emanating from the nozzles which at low pressures terminated in Mach disks and at higher pressures extended beyond the barrel shape and reflected off the walls of the duct. Based on pressure data from previous testing, reflected shocks were expected on the walls of the duct. The shadowgraph images confirmed the locations of these reflected shocks on the top wall of the duct. The shadowgraph images also showed the structure change which correlated to a change in pitch of the ejector noise, and corresponded to a change in trend of the duct wall pressure ratio distributions. The images produced from the setup provided insight into the complex flow behavior inside the ejector duct. In addition, the techniques were a valuable tool as an educational device for students

OSIRIS-REx, Returning the Asteroid Sample

This paper addresses the technical aspects of the sample return system for the upcoming Origins, Spectral Interpretation, Resource Identification, and Security-Regolith Explorer (OSIRIS-REx) asteroid sample return mission. The overall mission design and current implementation are presented as an overview to establish a context for the technical description of the reentry and landing segment of the mission.The prime objective of the OSIRIS-REx mission is to sample a primitive, carbonaceous asteroid and to return that sample to Earth in pristine condition for detailed laboratory analysis. Targeting the near-Earth asteroid Bennu, the mission launches in September 2016 with an Earth reentry date of September 24, 2023.OSIRIS-REx will thoroughly characterize asteroid Bennu providing knowledge of the nature of near-Earth asteroids that is fundamental to understanding planet formation and the origin of life. The return to Earth of pristine samples with known geologic context will enable precise analyses that cannot be duplicated by spacecraft-based instruments, revolutionizing our understanding of the early Solar System. Bennu is both the most accessible carbonaceous asteroid and one of the most potentially Earth-hazardous asteroids known. Study of Bennu addresses multiple NASA objectives to understand the origin of the Solar System and the origin of life and will provide a greater understanding of both the hazards and resources in near-Earth space, serving as a precursor to future human missions to asteroids.This paper focuses on the technical aspects of the Sample Return Capsule (SRC) design and concept of operations, including trajectory design and reentry retrieval. Highlights of the mission are included below.The OSIRIS-REx spacecraft provides the essential functions for an asteroid characterization and sample return mission: attitude control propulsion power thermal control telecommunications command and data handling structural support to ensure successful rendezvous with Bennu characterization of Bennus properties delivery of the sampler to the surface, and return of the spacecraft to the vicinity of the Earth sample collection, performed by the Touch-and-Go Sample Acquisition Mechanism (TAGSAM), to acquire a regolith sample from the surface Earth re-entry and SRC recovery. Following sample collection, OSIRIS-REx drifts away from Bennu until the Asteroid Departure Maneuver is commanded on March 4, 2021, sending OSIRIS-REx on a ballistic return cruise to Earth. No additional large deterministic maneuvers are required to return the SRC to Earth. During the cruise, tracking and trajectory correction maneuvers (TCMs) are performed as necessary to precisely target the entry corridor. As OSIRIS-REx approaches Earth, the reentry plans are reviewed starting about a year before arrival, and preparations begin. The spacecraft is targeted away from the Earth until 7 days before entry. The final two trajectory correction maneuvers bring the spacecraft on target toward the Utah Test and Training Range (UTTR), with sufficient time for contingency resolution. The SRC releases 4 hours prior to atmospheric entry interface and, using the Stardust capsule heritage design, employs a traditional drogue and main parachute descent system for a soft touchdown

Persistent SARS-CoV-2 PCR Positivity Despite Anti-viral Treatment in Immunodeficient Patients

PURPOSE: COVID-19 infection in immunodeficient individuals can result in chronically poor health, persistent or relapsing SARS-CoV-2 PCR positivity, and long-term infectious potential. While clinical trials have demonstrated promising outcomes using anti-SARS-CoV-2 medicines in immunocompetent hosts, their ability to achieve sustained viral clearance in immunodeficient patients remains unknown. We therefore aimed to study long-term virological outcomes in patients treated at our centre. METHODS: We followed up immunocompromised inpatients treated with casirivimab-imdevimab (Ronapreve) between September and December 2021, and immunocompromised patients who received sotrovimab, molnupiravir, nirmatrelvir/ritonavir (Paxlovid), or no treatment from December 2021 to March 2022. Nasopharyngeal swab and sputum samples were obtained either in hospital or in the community until sustained viral clearance, defined as 3 consecutive negative PCR samples, was achieved. Positive samples were sequenced and analysed for mutations of interest. RESULTS: We observed sustained viral clearance in 71 of 103 patients, none of whom died. Of the 32/103 patients where sustained clearance was not confirmed, 6 died (between 2 and 34 days from treatment). Notably, we observed 25 cases of sputum positivity despite negative nasopharyngeal swab samples, as well as recurrence of SARS-CoV-2 positivity following a negative sample in 12 cases. Patients were then divided into those who cleared within 28 days and those with PCR positivity beyond 28 days. We noted lower B cell counts in the group with persistent PCR positivity (mean (SD) 0.06 (0.10) ×109/L vs 0.22 (0.28) ×109/L, p = 0.015) as well as lower IgA (median (IQR) 0.00 (0.00-0.15) g/L vs 0.40 (0.00-0.95) g/L, p = 0.001) and IgM (median (IQR) 0.05 (0.00-0.28) g/L vs 0.35 (0.10-1.10) g/L, p = 0.005). No differences were seen in CD4+ or CD8+ T cell counts. Antiviral treatment did not impact risk of persistent PCR positivity. CONCLUSION: Persistent SARS-CoV-2 PCR positivity is common among immunodeficient individuals, especially those with antibody deficiencies, regardless of anti-viral treatment. Peripheral B cell count and serum IgA and IgM levels are predictors of viral persistence

Anomalous modes drive vortex dynamics in confined Bose-Einstein condensates

The dynamics of vortices in trapped Bose-Einstein condensates are investigated both analytically and numerically. In axially symmetric traps, the critical rotation frequency for the metastability of an isolated vortex coincides with the largest vortex precession frequency (or anomalous mode) in the Bogoliubov excitation spectrum. As the condensate becomes more elongated, the number of anomalous modes increases. The largest frequency of these modes exceeds both the thermodynamic critical frequency and the nucleation frequency at which vortices are created dynamically. Thus, anomalous modes describe not only the critical rotation frequency for creation of the first vortex in an elongated condensate but also the vortex precession in a single-component spherical condensate.Comment: 4 pages revtex, 3 embedded figure