ALBERT

Description: Many colorectal cancers have high levels of cyclo-oxygenase 2 (COX-2), an enzyme that metabolizes the essential fatty acids into prostaglandins. Since the low-density lipoprotein receptor (LDLr) is involved in the uptake of essential fatty acids, we studied the effect of LDL on growth and gene regulation in colorectal cancer cells. DiFi cells grown in lipoprotein-deficient sera (LPDS) grew more slowly than cells with LDL. LDLr antibody caused significant inhibition of tumor cell growth but did not affect controls. In addition, LDL uptake did not change in the presence of excess LDL, suggesting that ldlr mRNA lacks normal feedback regulation in some colorectal cancers. Analysis of the ldlr mRNA showed that excess LDL in the medium did not cause down-regulation of the message even after 24 hr. The second portion of the study examined the mRNA expression of ldlr and its co-regulation with cox-2 in normal and tumor specimens from patients with colorectal adenocarcinomas. The ratio of tumor:paired normal mucosa of mRNA expression of ldlr and of cox-2 was measured in specimens taken during colonoscopy. ldlr and cox-2 transcripts were apparent in 11 of 11 carcinomas. There was significant coordinate up-regulation both of ldlr and of cox-2 in 6 of 11 (55%) tumors compared with normal colonic mucosa. There was no up-regulation of cox-2 without concomitant up-regulation of ldlr. These data suggest that the LDLr is abnormally regulated in some colorectal tumors and may play a role in the up-regulation of cox-2. Copyright 1999 Wiley-Liss, Inc.

Description: Serum-deprived mouse osteoblastic (MC3T3E1) cells were subjected to a vibrational force modeled by NASA to simulate a space shuttle launch (7.83 G rms). The mRNA levels for eight genes were investigated to determine the effect of vibrational force on mRNA expression. The mRNA levels of two growth-related protooncogenes, c-fos and c-myc, were up-regulated significantly within 30 min after vibration, whereas those of osteocalcin as well as transforming growth factor-beta1 were decreased significantly within 3 h after vibration. No changes were detected in the levels of beta-actin, histone H4, or cytoplasmic phospholipase A2 after vibration. No basal levels of cyclooxygenase-2 expression were detected. In addition, the extracellular concentrations of prostaglandin E2 (PGE2), a potent autocrine/paracrine growth factor in bone, were not significantly altered after vibration most likely due to the serum deprivation state of the osteoblasts. In comparison with the gravitational launch profile, vibrational-induced changes in gene expression were greater both in magnitude and number of genes activated. Taken together, these data suggest that the changes in mRNA expression are due to a direct mechanical effect of the vibrational force on the osteoblast cells and not to changes in the local PGE2 concentrations. The finding that launch forces induce gene expression is of utmost importance since many of the biological experiments do not dampen vibrational loads on experimental samples. This lack of dampening of vibrational forces may partially explain why 1-G onboard controls sometimes do not reflect 1-G ground controls. These data may also suggest that scientists use extra ground controls that are exposed to launch forces, have these forces dampened on launched samples, or use facilities such as Biorack that provide an onboard 1-G centrufuge in order to control for space shuttle launch forces.

Description: No abstract available

Description: The well-defined osteoblast line, MC3T3-E1 was used to examine fibronectin (FN) mRNA levels, protein synthesis, and extracellular FN matrix accumulation after growth activation in spaceflight. These osteoblasts produce FN extracellular matrix (ECM) known to regulate adhesion, differentiation, and function in adherent cells. Changes in bone ECM and osteoblast cell shape occur in spaceflight. To determine whether altered FN matrix is a factor in causing these changes in spaceflight, quiescent osteoblasts were launched into microgravity and were then sera activated with and without a 1-gravity field. Synthesis of FN mRNA, protein, and matrix were measured after activation in microgravity. FN mRNA synthesis is significantly reduced in microgravity (0-G) when compared to ground (GR) osteoblasts flown in a centrifuge simulating earth's gravity (1-G) field 2.5 h after activation. However, 27.5 h after activation there were no significant differences in mRNA synthesis. A small but significant reduction of FN protein was found in the 0-G samples 2.5 h after activation. Total FN protein 27.5 h after activation showed no significant difference between any of the gravity conditions, however, there was a fourfold increase in absolute amount of protein synthesized during the incubation. Using immunofluorescence, we found no significant differences in the amount or in the orientation of the FN matrix after 27.5 h in microgravity. These results demonstrate that FN is made by sera-activated osteoblasts even during exposure to microgravity. These data also suggest that after a total period of 43 h of spaceflight FN transcription, translation, or altered matrix assembly is not responsible for the altered cell shape or altered matrix formation of osteoblasts.

Description: In serum-deprived MC3T3-E1 osteoblasts, mechanical stimulation caused by mild (287 x g) centrifugation induced a 10-fold increase in mRNA levels of the proto-oncogene, c-fos. Induction of c-fos was abolished by the cAMP-dependent protein kinase inhibitor H-89, suggesting that the transient c-fos mRNA increase is mediated by cAMP. Down-regulation of protein kinase C (PKC) activity by chronic TPA treatment failed to significantly reduce c-fos induction, suggesting that TPA-sensitive isoforms of PKC are not responsible for c-fos up-regulation. In addition, 287 x g centrifugation increased intracellular prostaglandin E2 (PGE2) levels 2.8-fold (P〈0. 005). Since we have previously shown that prostaglandin E2 (PGE2) can induce c-fos expression via a cAMP-mediated mechanism, we asked whether the increase in c-fos mRNA was due to centrifugation-induced PGE2 release. Pretreatment with the cyclooxygenase inhibitors indomethacin and flurbiprofen did not hinder the early induction of c-fos by mechanical stimulation. We conclude that c-fos expression induced by mild mechanical loading is dependent primarily on cAMP, not PKC, and initial induction of c-fos is not necessarily dependent on the action of newly synthesized PGE2.

Description: The development of prostate cancer has been linked to high level of dietary fat intake. Our laboratory investigates the connection between cancer cell growth and fatty acid products. Studying human prostatic carcinoma PC-3 cells, we found that prostaglandin E2 (PGE2) increased cell growth and up-regulated the gene expression of its own synthesizing enzyme, cyclooxygenase-2 (COX-2). PGE2 increased COX-2 mRNA expression dose-dependently with the highest levels of stimulation seen at the 3-hour period following PGE2 addition. The NSAID flurbiprofen (5 microM), in the presence of exogenous PGE2, inhibited the up-regulation of COX-2 mRNA and cell growth. These data suggest that the levels of local intracellular PGE2 play a major role in the growth of prostate cancer cells through an activation of COX-2 gene expression.

Description: Maintaining contamination certification of multi-mission flight hardware is an innovative approach to controlling mission costs. Methods for assessing ground induced degradation between missions have been employed by the Hubble Space Telescope (HST) Project for the multi-mission (servicing) hardware. By maintaining the cleanliness of the hardware between missions, and by controlling the materials added to the hardware during modification and refurbishment both project funding for contamination recertification and schedule have been significantly reduced. These methods will be discussed and HST hardware data will be presented.

Description: We present our first results from a study of the supernova remnants (SNRs) in the Large Magellanic Cloud (LMC) using data from ASCA. The three remnants we have analyzed to date, 0509-67.5, 0519-69.0, and N103B, are among the smallest, and presumably also the youngest, in the Cloud. The X-ray spectra of these SNRs show strong K(alpha) emission lines of silicon, sulfur, argon, and calcium with no evidence for corresponding lines of oxygen, neon, or magnesium. The dominant feature in the spectra is a broad blend of emission lines around 1 keV which we attribute to L-shell emission lines of iron. Model calculations (Nomoto, Thielemann, & Yokoi 1984) show that the major products of nucleosynthesis in Type Ia supernovae (SNs) are the elements from silicon to iron, as observed here. The calculated nucleosynthetic yields from Type Ib and II SNs are shown to be qualitatively inconsistent with the data. We conclude that the SNs which produced these remnants were of Type Ia. This finding also confirms earlier suggestions that the class of Balmer-dominated remnants arise from Type Ia SN explosions. Based an these early results from the LMC SNR sample, we find that roughly one-half of the SNRs produced in the LMC within the last approximately 1500 yr came from Type Ia SNs.

Description: The supernova remnant G292.0+1.8 was observed by the ROSAT PSPC for 18 ksec as part of this grant. Considerable effort was put into the analysis of the PSPC spectra. The major work went into nonequilibrium ionization joint spectral fits with the Einstein SSS and EXOSAT ME data which indicated that the two spatial regions of this remnant (a central bar and a plateau region covering a larger extent) had similar abundances, but different excitation conditions (temperature and ionization state), an important conclusion, if true. Unfortunately as this work was being finished, new ASCA data revealed the presence of a previously unknown, spectrally hard X-ray source near the center of the remnant which contaminated the SSS and ME data and as a consequence made our detailed spectral analysis done up until then un-publishable. We searched for evidence of this hard source in the PSPC data both spectrally and using timing searches (for a pulsar), but found nothing significant. ROSAT HRI data were also obtained on this remnant. These data were compared to the Einstein HRI data to search for evidence of spectral variations with position and possible expansion of the X-ray remnant. One feature in the remnant appears to have changed in brightness although it is not clear what is the cause of the change. No evidence for the hard ASCA source was apparent in the HRI data.

Description: We report the detection, using data from the Advanced Satellite for Cosmology and Astrophysics (ASCA), of a hard X-ray source in the vicinity of the radio pulsar PSR B1853+01, which is located within the supernova remnant (SNR) W44. PSR B1853+01, a 267 ms pulsar, has to date been detected only in the radio band. Previous observations at soft X-ray energies (e.g., with ROSAT HRI) have failed to detect any significant X-ray emission (pulsed or unpulsed) from the pulsar. In addition, no high-energy emission (approx. 〉 4 keV) has been detected previously from W44. Over the 0.5-4.0 keV band, the ASCA data show soft thermal emission from W44, with a morphology very similar to that observed earlier by Einstein and ROSAT. In the high-energy band (4.0-9.5 keV), the SNR is, for the most part, invisible, although a source coincident with the position of PSR B1853+01 is evident. The observed ASCA spectra are consistent with a power-law origin (photon index approx. 2.3) for the X-ray emission from this source at a flux level (flux density approx. 0.5 micro Jy at I keV) consistent with previous upper limits. The maximum allowed size for the source is determined directiv from the ASCA data (〈5 min.), while the minimum size is derived from the nondetection of a point source in the ROSAT HRI data (approx. 〉 30 sec.). Timing analysis of the hard X-ray source failed to detect pulsations at the pulsar's period. Based on these lines of evidence, we conclude that the new hard source in W44 represents an X-ray synchrotron nebula associated with PSR B1853+01, rather than the beamed output of the pulsar itself. This discoverv adds W44 to the small group of previously known plerionic SNRs This nebula lies at the low end of, but is consistent with, the correlation between X-ray luminosity and pulsar spin-down energy loss found for such objects, lending further support to our interpretation.