Study of Rat Lung Alveoli using Corrosion Casting and Freeze Fracture Methods Coupled with Digital Image Analysis

Relative areas and volumes can be estimated from vascular corrosion casts of rat lung alveoli using a calibration obtained from bulk frozen hydrated tissue. These morphometric measurements are roughly independent of the shrinkage and distortion artifacts known to arise in the corrosion casting procedure. Digital image processing of the SEM micrographs is employed to facilitate the measurement of casts and frozen tissue. The vascular corrosion casting technique is modified also to permit successful casting of alveolar air passages. The modified technique produces faithful casts of dead-ended luminal structures where continuous perfusion of casting medium into the tissue is not possible. The casts of alveolar air passages and their corresponding vasculature are compared to determine the volume of a single alveolus. By utilizing the calibrated measurements of lung areas and volumes, an estimate of lung vascular surface area per unit volume is obtained. This number, when multiplied by the respiratory tidal volume of the rat, may represent the total lung vascular surface area available for physiological gas exchange during normal respiration

Theory for Calcium-Phosphate Crystal Formation in Tissue from Scanning Electron Microscope Data

Scanning electron microscope (SEM) morphological analysis combined with energy dispersive characteristic x-ray analysis provides insight into the mechanism of biological mineralization. A time series of tissue micrographs and mineralization measurements can permit the determination of the mineralization kinetic behavior and is the basis upon which a computer model has been devised. The computer model is constructed from fundamental principles of crystal nucleation and precipitation theory. Various general forms of the model are tested against the laboratory data for goodness-of-fit using the least squares method, and two models are found to be acceptable. Both of the acceptable models involve inhibition of the mineralization process which has a reaction order ranging from one to two. A third model involving constant nucleation rate must be rejected. Having established working first principle models for the mineralization process, one can compute a constant number of nucleation sites and a supersaturation value for calcium in various mineralized tissues such as the spongiosa and fibrosa of heart valve leaflet implants. These quantities are determined and used in discussing a general theory for biomineralization which emphasizes therapeutic considerations

Scanning Electron Microscopy Methodology for Study of the Pathophysiology of Calcification in Bioprosthetic Heart Valves

Scanning electron microscope (SEM) morphologic analysis combined with energy dispersive characteristic X-ray (EDX) microprobe analysis provides insight into the mechanisms associated with disease-related crystal formation in biological materials. SEM and EDX were employed in analyzing specimens which were embedded in standard fashion in glycolmethacrylate (JB-4). The specimen surfaces under electron microscope investigation resulted from microtomy used in the preparation of reference light microscope histological sections; thus histology served as a direct reference for the SEM and EDX analyses. The particular application of these methods was in the study of bioprosthetic heart valve calcification, largely responsible for clinical failure of these heart valve substitutes. To simulate the clinically observed mineralization processes, glutaraldehyde-pretreated porcine heart valve leaflets were implanted subcutaneously in rats and subsequently removed at various time intervals from 1 to 56 days. Also, to address the hypothesis that the calcification process generates crystalline materials analogous to those in bone, EDX data obtained from pure hydroxyapatite were compared with the embedded tissue results. Further, EDX results were compared with data obtained by chemical analysis of the bulk specimens to assess the validity of the electron microscope technique

Use of Denosumab in Children With Osteoclast Bone Dysplasias: Report of Three Cases.

Denosumab has been used successfully to treat disease-associated osteoclast overactivity, including giant cell tumor of bone. Given its mechanism of action, denosumab is a potent potential treatment of other osteoclast bone dysplasias including central giant cell granuloma (CGCG), aneurysmal bone cyst (ABC), and cherubism. Relatively little is known about the safety and efficacy of denosumab in patients with these conditions, especially in children. We report on 3 pediatric patients treated with denosumab over a 3-year period at UCLA Medical Center (Los Angeles and Santa Monica, CA, USA): a 12-year-old with recurrent ABC of the pelvis, a 14-year-old with CGCG of the mandible, and a 12-year-old with cherubism. All were started on a 1-year course of 15 doses 120 mg s.c., given monthly with two loading doses on day 8 and 15. All patients demonstrated rapid and pronounced clinical improvement while on denosumab, including a significant reduction in pain and sclerosis of lytic lesions on radiographs. Within 1 month of initiating therapy, 2 patients experienced hypocalcemia (Common Terminology Criteria for Adverse Events [CTCAE] grade 2) and hypophosphatemia, with 1 patient experiencing symptoms. One patient went on to experience symptomatic rebound hypercalcemia (CTCAE grade 4) 5 months after completing therapy, requiring bisphosphonates and calcitonin. For the second patient, we developed a schedule to wean denosumab involving the progressive lengthening of time between doses from 1 to 4 months in 1-month increments before cessation. We found that denosumab therapy results in significant clinical and radiographic improvement for pediatric patients with nonresectable ABC, CGCG, and cherubism. Problems with serum calcium may be more common in younger patients, with symptomatic and protracted rebound hypercalcemia after cessation of therapy the most significant. We present a potential solution to this problem with progressive spacing of doses. Potential serious adverse events from alterations in calcium homeostasis should be explored in prospective clinical trials. © 2019 The Authors. JBMR Plus published by Wiley Periodicals, Inc. on behalf of American Society for Bone and Mineral Research

Shock Generation and Control Using DBD Plasma Actuators

This report is the final report of a NASA Phase I SBIR contract, with some revisions to remove company proprietary data. The Shock Boundary Layer Interaction (SBLI) phenomena in a supersonic inlet involve mutual interaction of oblique shocks with boundary layers, forcing the boundary layer to separate from the inlet wall. To improve the inlet efficiency, it is desired to prevent or delay shock-induced boundary layer separation. In this effort, Innovative Technology Applications Company (ITAC), LLC and the University of Notre Dame (UND) jointly investigated the use of dielectric-barrier-discharge (DBD) plasma actuators for control of SBLI in a supersonic inlet. The research investigated the potential for DBD plasma actuators to suppress flow separation caused by a shock in a turbulent boundary layer. The research involved both numerical and experimental investigations of plasma flow control for a few different SBLI configurations: (a) a 12 wedge flow test case at Mach 1.5 (numerical and experimental), (b) an impinging shock test case at Mach 1.5 using an airfoil as a shock generator (numerical and experimental), and (c) a Mach 2.0 nozzle flow case in a simulated 15 X 15 cm wind tunnel with a shock generator (numerical). Numerical studies were performed for all three test cases to examine the feasibility of plasma flow control concepts. These results were used to guide the wind tunnel experiments conducted on the Mach 1.5 12 degree wedge flow (case a) and the Mach 1.5 impinging shock test case (case b) which were at similar flow conditions as the corresponding numerical studies to obtain experimental evidence of plasma control effects for SBLI control. The experiments also generated data that were used in validating the numerical studies for the baseline cases (without plasma actuators). The experiments were conducted in a Mach 1.5 test section in the University of Notre Dame Hessert Laboratory. The simulation results from cases a and b indicated that multiple spanwise actuators in series and at a voltage of 75 kVp-p could fully suppress the flow separation downstream of the shock. The simulation results from case c showed that the streamwise plasma actuators are highly effective in creating pairs of counter-rotating vortices, much like the mechanical vortex generators, and could also potentially have beneficial effects for SBLI control. However, to achieve these effects, the positioning and the quantity of the DBD actuators used must be optimized. The wind tunnel experiments mapped the baseline flow with good agreement to the numerical simulations. The experimental results were conducted with spanwise actuators for cases a and b, but were limited by the inability to generate a sufficiently high voltage due to arcing in the wind-tunnel test-section. The static pressure in the tunnel was lower than the static pressure in an inlet at flight conditions, promoting arching and degrading the actuator performance

The survey of planetary nebulae in Andromeda (M31) III. Constraints from deep planetary nebula luminosity functions on the origin of the inner halo substructures in M31

The Andromeda (M31) galaxy displays several substructures in its inner halo whose origin as remnants of accreted satellites or perturbations of the pre-existing disc are encoded in the properties of their stellar populations (SPs), leaving traces on their deep [OIII] 5007 \AA planetary nebulae luminosity functions (PNLFs). By characterizing the morphology of the PNLFs, we constrain their origin. From our 54 sq. deg. deep narrow-band [OIII] survey of M31, we identify planetary nebulae (PNe) in the M31 disc and six major inner-halo substructures - the Giant Stream, North East Shelf, G1-Clump, Northern Clump, Western Shelf and Stream-D. We measure PNLF parameters from cumulative fits and statistically compare the PNLFs in each substructure and the disc. We link the PNLF parameters and those for the Large Magellanic Cloud to published metallicities and age measurements for their parent SPs. The absolute magnitudes of the PNLF bright cut-off ($M^{*}$) for these sub-populations span a significant magnitude range, despite having similar distance and line-of-sight extinction. $M^{*}$ for the Giant Stream, W-shelf and Stream-D PNLFs are fainter than those predicted by PN evolution models for the metallicity of the parent SPs. The faint-end slope of the PNLF increases linearly with decreasing fraction of stellar mass younger than 5 Gyr across the M31 regions and the LMC. From their PNLFs, the Giant Stream and NE-shelf are consistent with being stellar debris from an infalling satellite, while the G1 Clump appears to be linked with the pre-merger disc. The SPs of the substructures are consistent with those predicted by simulations of a single massive merger event that took place 2--3 Gyr ago in M31. Stream-D has an unrelated, distinct, origin. Furthermore, this study provides independent evidence that the faint-end of the PNLF is preferentially populated by PNe evolved from older stars.Comment: 14 pages, 7 figures, 4 tables; Abstract abridged; Accepted for publication at Astronomy & Astrophysic

Differences in Relative Hippocampus Volume and Number of Hippocampus Neurons among Five Corvid Species

The relative size of the avian hippocampus (Hp) has been shown to be related to spatial memory and food storing in two avian families, the parids and corvids. Basil et al. [Brain Behav Evol 1996;47: 156-164] examined North American food-storing birds in the corvid family and found that Clark’s nutcrackers had a larger relative Hp than pinyon jays and Western scrub jays. These results correlated with the nutcracker’s better performance on most spatial memory tasks and their strong reliance on stored food in the wild. However, Pravosudov and de Kort [Brain Behav Evol 67 (2006), 1-9] raised questions about the methodology used in the 1996 study, specifically the use of paraffin as an embedding material and recalculation for shrinkage. Therefore, we measured relative Hp volume using gelatin as the embedding material in four North American species of food-storing corvids (Clark’s nutcrackers, pinyon jays, Western scrub jays and blue jays) and one Eurasian corvid that stores little to no food (azure-winged magpies). Although there was a significant overall effect of species on relative Hp volume among the five species, subsequent tests found only one pairwise difference, blue jays having a larger Hp than the azure-winged magpies. We also examined the relative size of the septum in the five species. Although Shiflett et al. [J Neurobiol 51 (2002), 215-222] found a difference in relative septum volume amongst three species of parids that correlated with storing food, we did not find significant differences amongst the five species in relative septum. Finally, we calculated the number of neurons in the Hp relative to body mass in the five species and found statistically significant differences, some of which are in accord with the adaptive specialization hypothesis and some are not

RNA-Seq identifies genes whose proteins are upregulated during syncytia development in murine C2C12 myoblasts and human BeWo trophoblasts

The fusion of villous cytotrophoblasts into the multinucleated syncytiotrophoblast is critical for the essential functions of the mammalian placenta. Using RNA-Seq gene expression, quantitative protein expression, and siRNA knockdown we identified genes and their cognate proteins which are similarly upregulated in two cellular models of mammalian syncytia development (human BeWo cytotrophoblast to syncytiotrophoblast and murine C2C12 myoblast to myotube). These include DYSF, PDE4DIP, SPIRE2, NDRG1, PLEC, GPR146, HSPB8, DHCR7, and HDAC5. These findings provide avenues for further understanding of the mechanisms underlying mammalian placental syncytiotrophoblast development

Differences in Relative Hippocampus Volume and Number of Hippocampus Neurons among Five Corvid Species

The relative size of the avian hippocampus (Hp) has been shown to be related to spatial memory and food storing in two avian families, the parids and corvids. Basil et al. [Brain Behav Evol 1996;47: 156-164] examined North American food-storing birds in the corvid family and found that Clark’s nutcrackers had a larger relative Hp than pinyon jays and Western scrub jays. These results correlated with the nutcracker’s better performance on most spatial memory tasks and their strong reliance on stored food in the wild. However, Pravosudov and de Kort [Brain Behav Evol 67 (2006), 1-9] raised questions about the methodology used in the 1996 study, specifically the use of paraffin as an embedding material and recalculation for shrinkage. Therefore, we measured relative Hp volume using gelatin as the embedding material in four North American species of food-storing corvids (Clark’s nutcrackers, pinyon jays, Western scrub jays and blue jays) and one Eurasian corvid that stores little to no food (azure-winged magpies). Although there was a significant overall effect of species on relative Hp volume among the five species, subsequent tests found only one pairwise difference, blue jays having a larger Hp than the azure-winged magpies. We also examined the relative size of the septum in the five species. Although Shiflett et al. [J Neurobiol 51 (2002), 215-222] found a difference in relative septum volume amongst three species of parids that correlated with storing food, we did not find significant differences amongst the five species in relative septum. Finally, we calculated the number of neurons in the Hp relative to body mass in the five species and found statistically significant differences, some of which are in accord with the adaptive specialization hypothesis and some are not