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Modeling 238U/204Pb (μ) and Initial Pb of Lunar Meteorite Northwest Africa 12593

Apatite Ca 5 (PO 4 ) 3 (F, Cl, OH) serves as a valuable analytical resource. It is the only nominally hydrous phase on the moon and possesses the capacity to concentrate elements like U, Th, Pb, and rare earth elements (REE). This characteristic facilitates its use in U-Pb and Pb-Pb geochronology, allowing for the determination of the initial isotopic Pb and the -238 U/ -204 Pb ratio (μ) of the parent melt. In situ analysis of apatite grains can provide a detailed record of geochemical changes, revealing the broader petrogenetic and volatile history of the source material. This mineral is particularly informative in understanding the developmental history of its source. In situ analyses are crucial for lunar samples, predominantly composed of breccias, where correlating distinct grains with whole rock analyses poses challenges. This study utilizes in situ apatite isotopic analysis of apatite grains in lunar meteorite Northwest Africa 12593 and model the initial Pb isotopic composition and -238 U/ -204 Pb (μ) of the source. We present in situ Secondary Ionization Mass Spectrometry (SIMS) Pb-Pb analyses of 12 apatite grains in Northwest Africa (NWA) 12593 to test two hypotheses: 1) NWA 12593 clasts and matrix are closely related and from the same source, and 2) the matrix material previously documented as having a highland affinity has a lower modeled μ then KREEP.

K M Lehman Franco↗
NASA NTRS1987

Theoretical study of the dissociation energy and the red and violet band systems of CN

The dissociation energy (D sub O) of CN is determined to be 7.65 + or - 0.06 eV. This corresponds to delta H sub f (CN) = 105.3 + or - 1.5 kcal/mole, in excellent agreement with Engleman and Rouse (1975), but considerably larger than the recent value deduced from shock-tube studies by Colket (1984). The result is obtained not only from extensive ab initio MRCI calculations using a very large Gaussian basis set, but also from extrapolation of the directly computed value by comparison of computed and experimental results fo NO, C2, and N2. As an additional calibration of the methods, the D sub O value for CN was computed from the corresponding value for CN(-) using the experimental electron affinity data. The lifetime of the nu prime = 0 level of the violet (B 2 sigma + yields X 2 sigma +) system was computed to be 62.4 ns, in good agreement with both experiment and previous calculations. Lifetimes for the red (A 2 pi yields X 2 sigma +) system decrease with increasing nu prime, which is consistent both with the recent experiment and calculations. While the computed lifetimes are significantly longer that those obtained from the experiment, they are shorter than those deduced from an analysis of the solar spectrum. However the D sub O and f (sub OO) are consistent with Lambert's model for the solar spectrum.

Bauschlicher, Charles W., Jr.↗
NASA NTRS1988

Petrology and provenance of Apollo 15 drive tube 15007/8

The petrology of submillimeter fractions of soils from 13 levels of Apollo 15 drive tube 15007/8 have been studied through grain size analysis, modal analysis of the 90-150 micron sieve fraction, and electron probe microanalysis of monomineralic fragments of olivine, pyroxene, and plagioclase from the soils. It is shown that core 15007/8 does not contain distinct lithologic units and probably contains soils of mixed maturity in which mixing has greatly dominated in situ reworking. No petrologic layering is found in the core. Abundances of KREEP basalts correlate with mare basalts and not with anorthosite, norite, and troctolite components from the Apennine Front. Some olivine and pyroxene fragments with mare basalt affinity are found to be optically and chemically unzoned, suggesting that they may have been derived from unusual source rocks.

Basu, Abhijit↗
NASA NTRS1991

Characterization of atrial natriuretic peptide receptors in brain microvessel endothelial cells

Atrial natriuretic peptide (ANP) binding and ANP-induced increases in cyclic guanosine monophosphate (cGMP) levels have been observed in brain microvessels (Chabrier et al., 1987; Steardo and Nathanson, 1987), suggesting that this fluid-regulating hormone may play a role in the fluid homeostasis of the brain. This study was initiated to characterize the ANP receptors in primary cultures of brain microvessel endothelial cells (BMECs). The apparent equilibrium dissociation constant, Kd, for ANP increased from 0.25 nM to 2.5 nM, and the number of ANP binding sites as determined by Scatchard analysis increased from 7,100 to 170,000 sites/cell between 2 and 10 days of culture following monolayer formation. Time- and concentration-dependent studies on the stimulation of cGMP levels by ANP indicated that guanylate cyclase-linked ANP receptors were present in BMECs. The relative abilities of ANP, brain natriuretic peptide (BNP), and a truncated analog of ANP containing amino acids 5-27 (ANP 5-27) to modulate the accumulation of cGMP was found to be ANP greater than BNP much greater than ANP 5-27. Affinity cross-linking with disuccinimidyl suberate and radiolabeled ANP followed by gel electrophoresis under reducing conditions demonstrated a single band corresponding to the 60-70 kD receptor, indicating the presence of the nonguanylate cyclase-linked ANP receptor. Radiolabeled ANP binding was examined in the presence of various concentrations of either ANP, BNP, or ANP 5-27 and suggested that a large proportion of the ANP receptors present in blood-brain barrier endothelial cells bind all of these ligands similarly. These data indicate both guanylate cyclase linked and nonguanylate cyclase linked receptors are present on BMECs and that a higher proportion of the nonguanylate cyclase linked receptors is expressed. This in vitro culture system may provide a valuable tool for the examination of ANP receptor expression and function in blood-brain barrier endothelial cells.

Endothelium, Vascular/chemistry↗
NASA NTRS1995

Application of Ni-63 photo and corona discharge ionization for the analysis of chemical warfare agents and toxic wastes

Over the past decade, advances in instrumental design and refinements in the understanding of ion molecule reactions at atmospheric pressure enabled the application of Ion Mobility Spectrometry (IMS) as a simple inexpensive and sensitive analytical method for the detection of organic trace compounds. Positive and negative gas-phase ions for ion mobility spectrometry have been produced by a variety of methods, including photo-ionization, laser multi photon ionization, surface ionization, corona discharge ionization. The most common ion source used in ion mobility spectrometry is a radioactive Ni-63 foil which is favored due to simplicity, stability, convenience, and high selectivity. If reactant ions like (H2O(n)H)(+) or (H2O(n)O2)(-) dominate in the reaction region, nearly all kinds of compounds with a given proton or electron affinity; are ionized. However, the radioactivity of the Ni-63 foil is one disadvantage of this ion source that stimulates the development and application of other ionization techniques. In this paper, we report analyses of old chemical warfare agents and toxic wastes using Bruker RAID ion mobility spectrometers. Due to the modular construction of the measuring cell, the spectrometers can be equipped with different ion sources. The combined use of Ni-63, photo- and corona discharge ionization allows the identification of different classes of chemical compounds and yields in most cases comparable results.

Stach, J.↗
NASA NTRS2001

Expression of the Acyl-Coenzyme A: Cholesterol Acyltransferase GFP Fusion Protein in Sf21 Insect Cells

The enzyme acyl-coenzyme A:cholesterol acyltransferase (ACAT) is an important contributor to the pathological expression of plaque leading to artherosclerosis n a major health problem. Adequate knowledge of the structure of this protein will enable pharmaceutical companies to design drugs specific to the enzyme. ACAT is a membrane protein located in the endoplasmic reticulum.t The protein has never been purified to homogeneity.T.Y. Chang's laboratory at Dartmouth College provided a 4-kb cDNA clone (K1) coding for a structural gene of the protein. We have modified the gene sequence and inserted the cDNA into the BioGreen His Baculovirus transfer vector. This was successfully expressed in Sf2l insect cells as a GFP-labeled ACAT protein. The advantage to this ACAT-GFP fusion protein (abbreviated GCAT) is that one can easily monitor its expression as a function of GFP excitation at 395 nm and emission at 509 nm. Moreover, the fusion protein GCAT can be detected on Western blots with the use of commercially available GFP antibodies. Antibodies against ACAT are not readily available. The presence of the 6xHis tag in the transfer vector facilitates purification of the recombinant protein since 6xHis fusion proteins bind with high affinity to Ni-NTA agarose. Obtaining highly pure protein in large quantities is essential for subsequent crystallization. The purified GCAT fusion protein can readily be cleaved into distinct GFP and ACAT proteins in the presence of thrombin. Thrombin digests the 6xHis tag linking the two protein sequences. Preliminary experiments have indicated that both GCAT and ACAT are expressed as functional proteins. The ultimate aim is to obtain large quantities of the ACAT protein in pure and functional form appropriate for protein crystal growth. Determining protein structure is the key to the design and development of effective drugs. X-ray analysis requires large homogeneous crystals that are difficult to obtain in the gravity environment of earth. Protein crystals grown in microgravity are often larger and have fewer defects than those grown on earth. The analysis of higher quality space-grown crystals will assist in structure-based drug design. We have successfully grown GCAT-infected Sf21 cells in both adhesion and suspension cultures. Expression levels of GCAT in cell lines such as Sf9 and High Five appear to be reduced. We intend to replicate GCAT expression in all three cell lines using the NASA rotating wall bioreactor which effectively duplicates a microgravity environment. The bioreactor itself could be launched to study the expression of the GFP and GCAT proteins in the actual microgravity environment achieved in orbit.

Mahtani, H. K.↗
NASA NTRS1985

A systems analysis of the erythropoietic responses to weightlessness. Volume 2: Description of the model of erythropoiesis regulation. Part A: Model for regulation of erythropoiesis. Part B: Detailed description of the model for regulation of erythropoiesis

A mathematical model of the erythropoiesis on total red blood cell mass is presented. The loss of red cell mass has been a consistent finding during space flight. Computer simulation of this phenomenon required a model that could account for oxygen transport, red cell production, and red cell destruction. The elements incorporated into the feedback regulation loop of the model are based on the accepted concept that erythrocyte production is governed by the balance between oxygen supply and demand in the body. The mechanisms and pathways of the control circuit include oxygenation of hemoglobin and oxygenation of tissues by blood transport and diffusional processes. Other features of the model include a variable oxygen-hemoglobin affinity, and time delays which represent time for erythropoietin (erythrocyte-stimulating hormone) distribution in plasma, and time for maturation of the erythrocytes in bone marrow.

Leonard, J. I.↗
NASA NTRS2014

Humic Acid Complexation of Th, Hf and Zr in Ligand Competition Experiments: Metal Loading and Ph Effects

The mobility of metals in soils and subsurface aquifers is strongly affected by sorption and complexation with dissolved organic matter, oxyhydroxides, clay minerals, and inorganic ligands. Humic substances (HS) are organic macromolecules with functional groups that have a strong affinity for binding metals, such as actinides. Thorium, often studied as an analog for tetravalent actinides, has also been shown to strongly associate with dissolved and colloidal HS in natural waters. The effects of HS on the mobilization dynamics of actinides are of particular interest in risk assessment of nuclear waste repositories. Here, we present conditional equilibrium binding constants (Kc, MHA) of thorium, hafnium, and zirconium-humic acid complexes from ligand competition experiments using capillary electrophoresis coupled with ICP-MS (CE- ICP-MS). Equilibrium dialysis ligand exchange (EDLE) experiments using size exclusion via a 1000 Damembrane were also performed to validate the CE-ICP-MS analysis. Experiments were performed at pH 3.5-7 with solutions containing one tetravalent metal (Th, Hf, or Zr), Elliot soil humic acid (EHA) or Pahokee peat humic acid (PHA), and EDTA. CE-ICP-MS and EDLE experiments yielded nearly identical binding constants for the metal- humic acid complexes, indicating that both methods are appropriate for examining metal speciation at conditions lower than neutral pH. We find that tetravalent metals form strong complexes with humic acids, with Kc, MHA several orders of magnitude above REE-humic complexes. Experiments were conducted at a range of dissolved HA concentrations to examine the effect of [HA]/[Th] molar ratio on Kc, MHA. At low metal loading conditions (i.e. elevated [HA]/[Th] ratios) the ThHA binding constant reached values that were not affected by the relative abundance of humic acid and thorium. The importance of [HA]/[Th] molar ratios on constraining the equilibrium of MHA complexation is apparent when our estimated Kc, MHA values attained at very low metal loading conditions are compared to existing literature data. Overall, experimental data suggest that the tetravalent transition metal/-actinide-humic acid complexation is important over a wide range of pH values, including mildly acidic conditions, and thus, these complexes should be included in speciation models.

humic acid↗
NASA NTRS1993

Characterization of the DNA binding properties of polyomavirus capsid protein

The DNA binding properties of the polyomavirus structural proteins VP1, VP2, and VP3 were studied by Southwestern analysis. The major viral structural protein VP1 and host-contributed histone proteins of polyomavirus virions were shown to exhibit DNA binding activity, but the minor capsid proteins VP2 and VP3 failed to bind DNA. The N-terminal first five amino acids (Ala-1 to Lys-5) were identified as the VP1 DNA binding domain by genetic and biochemical approaches. Wild-type VP1 expressed in Escherichia coli (RK1448) exhibited DNA binding activity, but the N-terminal truncated VP1 mutants (lacking Ala-1 to Lys-5 and Ala-1 to Cys-11) failed to bind DNA. The synthetic peptide (Ala-1 to Cys-11) was also shown to have an affinity for DNA binding. Site-directed mutagenesis of the VP1 gene showed that the point mutations at Pro-2, Lys-3, and Arg-4 on the VP1 molecule did not affect DNA binding properties but that the point mutation at Lys-5 drastically reduced DNA binding affinity. The N-terminal (Ala-1 to Lys-5) region of VP1 was found to be essential and specific for DNA binding, while the DNA appears to be non-sequence specific. The DNA binding domain and the nuclear localization signal are located in the same N-terminal region.

NASA Discipline Cell Biology↗
NASA NTRS2005

Multiple melanocortin receptors are expressed in bone cells

Melanocortin receptors belong to the seven transmembrane domain, G-protein coupled family of receptors. There are five members of this receptor family labeled MC1R-MC5R. These receptors are activated by fragments derived from a larger molecule, proopiomelanocortin (POMC) and include ACTH, alpha beta and gamma-MSH and beta-endorphin. Because of in vitro and in vivo data suggesting direct effects of these POMC molecules on bone and bone turnover, we examined bone and bone derived cells for the presence of the various members of the melanocortin receptor family. We report that the five known melanocortin receptors are expressed to varying degrees in osteoblast-like and osteoclastic cells. POMC fragments increased proliferation and expression of a variety of genes in osteoblastic cells. Furthermore, POMC mRNA was detected in osteoclastic cells. These data demonstrate that POMC-derived peptide hormones acting through high affinity melanocortin receptors have specific effects on bone cells. Thus, in addition to the indirect effects of POMC-derived hormones on bone turnover through their modulation of steroid hormone secretion, POMC fragments may have direct and specific effects on bone cell subpopulations.

NASA Discipline Musculoskeletal↗
NASA NTRS1999

Cloning the promoter for transforming growth factor-beta type III receptor. Basal and conditional expression in fetal rat osteoblasts

Transforming growth factor-beta binds to three high affinity cell surface molecules that directly or indirectly regulate its biological effects. The type III receptor (TRIII) is a proteoglycan that lacks significant intracellular signaling or enzymatic motifs but may facilitate transforming growth factor-beta binding to other receptors, stabilize multimeric receptor complexes, or segregate growth factor from activating receptors. Because various agents or events that regulate osteoblast function rapidly modulate TRIII expression, we cloned the 5' region of the rat TRIII gene to assess possible control elements. DNA fragments from this region directed high reporter gene expression in osteoblasts. Sequencing showed no consensus TATA or CCAAT boxes, whereas several nuclear factors binding sequences within the 3' region of the promoter co-mapped with multiple transcription initiation sites, DNase I footprints, gel mobility shift analysis, or loss of activity by deletion or mutation. An upstream enhancer was evident 5' proximal to nucleotide -979, and a silencer region occurred between nucleotides -2014 and -2194. Glucocorticoid sensitivity mapped between nucleotides -687 and -253, whereas bone morphogenetic protein 2 sensitivity co-mapped within the silencer region. Thus, the TRIII promoter contains cooperative basal elements and dispersed growth factor- and hormone-sensitive regulatory regions that can control TRIII expression by osteoblasts.

Non-NASA Center↗
NASA NTRS1996

Promoter-dependent and -independent activation of insulin-like growth factor binding protein-5 gene expression by prostaglandin E2 in primary rat osteoblasts

Insulin-like growth factor (IGF) action is mediated by high affinity cell surface IGF receptors and modulated by a family of secreted IGF binding proteins (IGFBPs). IGFBP-5, the most conserved of six IGFBPs characterized to date, uniquely potentiates the anabolic actions of IGF-I for skeletal cells. In osteoblasts, IGFBP-5 production is stimulated by prostaglandin E2 (PGE2), a local factor that mediates certain effects induced by parathyroid hormone, cytokines such as interleukin-1 and transforming growth factor-beta, and mechanical strain. In this study, we show that transcriptional and post-transcriptional events initiated by PGE2 collaborate to enhance IGFBP-5 gene expression in primary fetal rat osteoblast cultures. PGE2 treatment stimulated up to a 7-fold rise in steady-state levels of IGFBP-5 mRNA throughout 32 h of incubation. Analysis of nascent IGFBP-5 mRNA suggested that PGE2 had only a modest stimulatory effect on IGFBP-5 gene transcription, and transient transfection studies with IGFBP-5 promoter-reporter genes confirmed that PGE2 enhanced promoter activity by approximately 2-fold. Similar stimulatory effects were seen with forskolin. A DNA fragment with only 51 base pairs of the 5'-flanking sequence retained hormonal responsiveness, which may be mediated by a binding site for transcription factor AP-2 located at positions -44 to -36 in the proximal IGFBP-5 promoter. Incubation of osteoblasts with the mRNA transcriptional inhibitor 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole demonstrated that PGE2 enhanced IGFBP-5 mRNA stability by 2-fold, increasing the t1/2 from 9 to 18 h. The effects of PGE2 on steady-state IGFBP-5 transcripts were abrogated by preincubating cells with cycloheximide, indicating that the effects of PGE2 on both gene transcription and mRNA stability required ongoing protein synthesis. Therefore, both promoter-dependent and -independent pathways converge to enhance IGFBP-5 gene expression in response to PGE2 in osteoblasts.

Non-NASA Center↗
NASA NTRS2020

SAGE III ISS Contamination Monitoring Package: Observations in Orbit

The Stratospheric Aerosol and Gas Experiment III (SAGE III) telescope and instrument assembly employ the methods of solar occultation and lunar occultation to retrieve near-global vertical profiles of atmospheric ozone, water vapor, nitrogen dioxide, aerosol extinctions, and other gaseous species and atmospheric state parameters. The SAGE III grating spectrometer measures light within the spectral range of 280 nm to 1037 nm at approximately 1 nm resolution, but retrievals in the Ultraviolet (UV) are particularly sensitive to contamination of the optical train. Therefore, a contamination door that contains a quartz optical window can be closed over the telescope aperture during periods of enhanced external contaminant flux. This optically transparent window permits continued science event acquisition at an acceptably diminished signal-to-noise ratio, which is expected to decline with ongoing accretion of contaminant material. To date, this impact has been short term, and science quality through the window returns to baseline performance after a contamination source is removed and spontaneous desorption removes material from the low-affinity quartz surface. Two Contamination Monitoring Packages (CMPs) consisting of eight Thermoelectric Quartz Crystal Microbalances (TQCMs) from QCM Research provide characterization and redundant monitoring of contaminant deposition from the 2π steradian solid angle on the payload side of the Expedite the Processing of Experiments to the Space Station (ExPRESS) Payload Adapter. CMP data are closely examined by the SAGE III team to determine when the contamination door should remain closed during science events and in what direction the instrument assembly scan head should stow when not acquiring science measurements. Additionally, should the CMPs indicate an unacceptable accretion rate, the ight computer will close the contamination door as part of the automatic fault detection system. Along with spectrometer measurements of the quartz window's transmission, the payload CMPs enable auditing of the mission contamination budget. The process of Thermogravimetric Analysis (TGA) can be used to help identify chemical constituents accreted on the CMP sensors. To be presented here along with an explanation of the CMP systems are the first two and a half years of observations of the contaminant deposition environment around the payload in quiescence and during special events like docking vehicles.

Tyler T Dawson↗
NASA NTRS2020

SAGE III/ISS Contamination Monitoring Package: Observations in Orbit

The Stratospheric Aerosol and Gas Experiment III (SAGE III) telescope and instrument assembly employ the methods of solar occultation and lunar occultation to retrieve near-global vertical profiles of atmospheric ozone, water vapor, nitrogen dioxide, aerosol extinctions, and other gaseous species and atmospheric state parameters. The SAGE III grating spectrometer measures light within the spectral range of 280 nm to 1037 nm at approximately 1 nm resolution, but retrievals in the Ultraviolet (UV) are particularly sensitive to contamination of the optical train. Therefore, a contamination door that contains a quartz optical window can be closed over the telescope aperture during periods of enhanced external contaminant flux. This optically transparent window permits continued science event acquisition at an acceptably diminished signal-to-noise ratio, which is expected to decline with ongoing accretion of contaminant material. To date, this impact has been short term, and science quality through the window returns to baseline performance after a contamination source is removed and spontaneous desorption removes material from the low-affinity quartz surface. Two Contamination Monitoring Packages (CMPs) consisting of eight Thermoelectric Quartz Crystal Microbalances (TQCMs) from QCM Research provide characterization and redundant monitoring of contaminant deposition from the 2pi steradian solid angle on the payload side of the Expedite the Processing of Experiments to the Space Station (ExPRESS) Payload Adapter. CMP data are closely examined by the SAGE III team to determine when the contamination door should remain closed during science events and in what direction the instrument assembly scan head should stow when not acquiring science measurements. Additionally, should the CMPs indicate an unacceptable accretion rate, the flight computer will close the contamination door as part of the automatic fault detection system. Along with spectrometer measurements of the quartz window's transmission, the payload CMPs enable auditing of the mission contamination budget. The process of Thermogravimetric Analysis (TGA) can be used to help identify chemical constituents accreted on the CMP sensors. To be presented here along with an explanation of the CMP systems are the first two and a half years of observations of the contaminant deposition environment around the payload in quiescence and during special events like docking vehicles.

Tyler T Dawson↗
NASA NTRS1983

Generic buckling curves for specially orthotropic rectangular plates

Using a double affine transformation, the classical buckling equation for specially orthotropic plates and the corresponding virtual work theorem are presented in a particularly simple fashion. These dual representations are characterized by a single material constant, called the generalized rigidity ratio, whose range is predicted to be the closed interval from 0 to 1 (if this prediction is correct then the numerical results using a ratio greater than 1 in the specially orthotropic plate literature are incorrect); when natural boundary conditions are considered a generalized Poisson's ratio is introduced. Thus the buckling results are valid for any specially orthotropic material; hence the curves presented in the text are generic rather than specific. The solution trends are twofold; the buckling coefficients decrease with decreasing generalized rigidity ratio and, when applicable, they decrease with increasing generalized Poisson's ratio. Since the isotropic plate is one limiting case of the above analysis, it is also true that isotropic buckling coefficients decrease with increasing Poission's ratio.

Brunnelle, E. J.↗
NASA NTRS1988

Nature and origin of mineral coatings on volcanic rocks of the Black Mountain, Stonewall Mountain, and Kane Springs Wash volcanic centers, Southern Nevada

Comparative lab spectra and Thematic Mapper imagery investigations at 3 Tertiary calderas in southern Nevada indicate that desert varnish is absorbant relative to underlying host rocks below about 0.7 to 1.3 microns, depending on mafic affinity of the sample, but less absorbant than mafic host rocks at higher wavelengths. Desert varnish occurs chiefly as thin impregnating films. Distribution of significant varnish accumulations is sparse and localized, occurring chiefly in surface recesses. These relationships result in the longer wavelength bands and high 5/2 values over felsic units with extensive desert varnish coatings. These lithologic, petrochemical, and desert varnish controlled spectral responses lead to characteristic TM band relationships which tend to correlate with conventionally mappable geologic formations. The concept of a Rock-Varnish Index (RVI) is introduced to help distinguish rocks with a potentially detectable varnish. Felsic rocks have a high RVI, and those with extensive desert varnish behave differently, spectrally, from those without extensive varnish. The spectrally distinctive volcanic formations at Stonewall Mountain provide excellent statistical class segregation on supervised classification images. A binary decision rule flow-diagram is presented to aid TM imagery analysis over volcanic terrane in semi-arid environments.

Taranik, James V.↗
NASA NTRS1999

Collagenase-3 binds to a specific receptor and requires the low density lipoprotein receptor-related protein for internalization

We have previously identified a specific receptor for collagenase-3 that mediates the binding, internalization, and degradation of this ligand in UMR 106-01 rat osteoblastic osteosarcoma cells. In the present study, we show that collagenase-3 binding is calcium-dependent and occurs in a variety of cell types, including osteoblastic and fibroblastic cells. We also present evidence supporting a two-step mechanism of collagenase-3 binding and internalization involving both a specific collagenase-3 receptor and the low density lipoprotein receptor-related protein. Ligand blot analysis shows that (125)I-collagenase-3 binds specifically to two proteins ( approximately 170 kDa and approximately 600 kDa) present in UMR 106-01 cells. Western blotting identified the 600-kDa protein as the low density lipoprotein receptor-related protein. Our data suggest that the 170-kDa protein is a specific collagenase-3 receptor. Low density lipoprotein receptor-related protein-null mouse embryo fibroblasts bind but fail to internalize collagenase-3, whereas UMR 106-01 and wild-type mouse embryo fibroblasts bind and internalize collagenase-3. Internalization, but not binding, is inhibited by the 39-kDa receptor-associated protein. We conclude that the internalization of collagenase-3 requires the participation of the low density lipoprotein receptor-related protein and propose a model in which the cell surface interaction of this ligand requires a sequential contribution from two receptors, with the collagenase-3 receptor acting as a high affinity primary binding site and the low density lipoprotein receptor-related protein mediating internalization.

NASA Discipline Musculoskeletal↗
NASA NTRS2000

Quantitative image analysis of laminin immunoreactivity in skin basement membrane irradiated with 1 GeV/nucleon iron particles

We previously reported that laminin immunoreactivity in mouse mammary epithelium is altered shortly after whole-body irradiation with 0.8 Gy from 600 MeV/nucleon iron ions but is unaffected after exposure to sparsely ionizing radiation. This observation led us to propose that the effect could be due to protein damage from the high ionization density of the ion tracks. If so, we predicted that it would be evident soon after radiation exposure in basement membranes of other tissues and would depend on ion fluence. To test this hypothesis, we used immunofluorescence, confocal laser scanning microscopy, and image segmentation techniques to quantify changes in the basement membrane of mouse skin epidermis. At 1 h after exposure to 1 GeV/nucleon iron ions with doses from 0.03 to 1.6 Gy, neither the visual appearance nor the mean pixel intensity of laminin in the basement membrane of mouse dorsal skin epidermis was altered compared to sham-irradiated tissue. This result does not support the hypothesis that particle traversal directly affects laminin protein integrity. However, the mean pixel intensity of laminin immunoreactivity was significantly decreased in epidermal basement membrane at 48 and 96 h after exposure to 0.8 Gy 1 GeV/nucleon iron ions. We confirmed this effect with two additional antibodies raised against affinity-purified laminin 1 and the E3 fragment of the long-arm of laminin 1. In contrast, collagen type IV, another component of the basement membrane, was unaffected. Our studies demonstrate quantitatively that densely ionizing radiation elicits changes in skin microenvironments distinct from those induced by sparsely ionizing radiation. Such effects may might contribute to the carcinogenic potential of densely ionizing radiation by altering cellular signaling cascades mediated by cell-extracellular matrix interactions.

Non-NASA Center↗