Stable low-level expression of p21(WAF1/CIP1 )in A549 human bronchogenic carcinoma cell line-derived clones down-regulates E2F1 mRNA and restores cell proliferation control

BACKGROUND: Deregulated cell cycle progression and loss of proliferation control are key properties of malignant cells. In previous studies, an interactive transcript abundance index (ITAI) comprising three cell cycle control genes, [MYC × E2F1]/p21 accurately distinguished normal from malignant bronchial epithelial cells (BEC), using a cut-off threshold of 7,000. This cut-off is represented by a line with a slope of 7,000 on a bivariate plot of p21 versus [MYC × E2F1], with malignant BEC above the line and normal BEC below the line. This study was an effort to better quantify, at the transcript abundance level, the difference between normal and malignant BEC. The hypothesis was tested that experimental elevation of p21 in a malignant BEC line would decrease the value of the [MYC × E2F1]/p21 ITAI to a level below this line, resulting in loss of immortality and limited cell population doubling capacity. In order to test the hypothesis, a p21 expression vector was transfected into the A549 human bronchogenic carcinoma cell line, which has low constitutive p21 TA expression relative to normal BEC. RESULTS: Following transfection of p21, four A549/p21 clones with stable two-fold up-regulated p21 expression were isolated and expanded. For each clone, the increase in p21 transcript abundance (TA) was associated with increased total p21 protein level, more than 5-fold reduction in E2F1 TA, and 10-fold reduction in the [MYC × E2F1]/p21 ITAI to a value below the cut-off threshold. These changes in regulation of cell cycle control genes were associated with restoration of cell proliferation control. Specifically, each transfectant was capable of only 15 population doublings compared with unlimited population doublings for parental A549. This change was associated with an approximate 2-fold increase in population doubling time to 38.4 hours (from 22.3 hrs), resumption of contact-inhibition, and reduced dividing cell fraction as measured by flow cytometric DNA analysis. CONCLUSION: These results, likely due to increased p21-mediated down-regulation of E2F1 TA at the G1/S phase transition, are consistent with our hypothesis. Specifically, they provide experimental confirmation that a line with slope of 7,000 on the p21 versus [MYC × E2F1] bivariate plot quantifies the difference between normal and malignant BEC at the level of transcript abundance

Variation in transcriptional regulation of cyclin dependent kinase inhibitor p21(waf1/cip1 )among human bronchogenic carcinomas

BACKGROUND: Cell proliferation control depends in part on the carefully ordered regulation of transcription factors. The p53 homolog p73, contributes to this control by directly upregulating the cyclin dependent kinase inhibitor, p21(waf1/cip1). E2F1, an inducer of cell proliferation, directly upregulates p73 and in some systems upregulates p21 directly. Because of its central role in controlling cell proliferation, upregulation of p21 has been explored as a modality for treating bronchogenic carcinoma (BC). Improved understanding of p21 transcriptional regulation will facilitate identification of BC tissues that are responsive to p21-directed therapies. Toward this goal, we investigated the role that E2F1 and p73 each play in the transcriptional regulation of p21. RESULTS: Among BC samples (N = 21) p21 transcript abundance (TA) levels varied over two orders of magnitude with values ranging from 400 to 120,000 (in units of molecules/10(6 )molecules β-actin). The p21 values in many BC were high compared to those observed in normal bronchial epithelial cells (BEC) (N = 18). Among all BC samples, there was no correlation between E2F1 and p21 TA but there was positive correlation between E2F1 and p73α (p < 0.001) TA. Among BC cell lines with inactivated p53 and wild type p73 (N = 7) there was positive correlation between p73α and p21 TA (p < 0.05). Additionally, in a BC cell line in which both p53 and p73 were inactivated (H1155), E2F1 TA level was high (50,000), but p21 TA level was low (470). Transiently expressed exogenous p73α in the BC cell line Calu-1, was associated with a significant (p < 0.05) 90% increase in p21 TA and a 20% reduction in E2F1 TA. siRNA mediated reduction of p73 TA in the N417 BC cell line was associated with a significant reduction in p21 TA level (p < 0.01). CONCLUSION: p21 TA levels vary considerably among BC patients which may be attributable to 1) genetic alterations in Rb and p53 and 2) variation in TA levels of upstream transcription factors E2F1 and p73. Here we provide evidence that p73 upregulates p21 TA in BC tissues and upregulated p21 TA may result from E2F1 upregulation of p73 but not from E2F1 directly

TESS observations of the Pleiades cluster: a nursery for delta Scuti stars

We studied 89 A- and F-type members of the Pleiades open cluster, including five escaped members. We measured projected rotational velocities (v sin i) for 49 stars and confirmed that stellar rotation causes a broadening of the main sequence in the color-magnitude diagram. Using time-series photometry from NASA's TESS Mission (plus one star observed by Kepler/K2), we detected delta Scuti pulsations in 36 stars. The fraction of Pleiades stars in the middle of the instability strip that pulsate is unusually high (over 80%), and their range of effective temperatures agrees well with theoretical models. On the other hand, the characteristics of the pulsation spectra are varied and do not correlate with stellar temperature, calling into question the existence of a useful nu_max relation for delta Scutis, at least for young stars. By including delta Scuti stars observed in the Kepler field, we show that the instability strip is shifted to the red with increasing distance by interstellar reddening. Overall, this work demonstrates the power of combining observations with Gaia and TESS for studying pulsating stars in open clusters.Comment: submitted to AAS journal

Autonomic Nervous System Function Following Prenatal Opiate Exposure

In utero exposure to opiates may affect autonomic functioning of the fetus and newborn. We investigated heart rate variability (HRV) as a measure of autonomic stability in prenatal opiate-exposed neonates (n = 14) and in control term infants (n = 10). Electrocardiographic data during both non-nutritive and nutritive sucking were evaluated for RR intervals, heart rate (HR), standard deviation of the consecutive RR intervals (SDRR), standard deviation of the differences of consecutive RR intervals (SDDRR), and the power spectral densities in low and high frequency bands. In controls, mean HR increased significantly, 143-161 per min (p = 0.002), with a trend toward a decrease in RR intervals from non-nutritive to nutritive sucking; these measures did not change significantly among exposed infants. Compared to controls, exposed infants demonstrated significantly greater HRV or greater mean SDRR and SDDRR during non-nutritive period (p \u3c 0.01), greater mean SDDRR during nutritive sucking (p = 0.02), and higher powers in the low and high frequency bands during nutritive feedings. Our findings suggest that prenatal opiate exposure may be associated with changes in autonomic nervous system (ANS) functioning involving both sympathetic and parasympathetic branches. Future studies are needed to examine the effects of prenatal opiate exposure on ANS function

Elevated DNA Oxidation and DNA Repair Enzyme Expression in Brain White Matter in Major Depressive Disorder

Background: Pathology of white matter in brains of patients with major depressive disorder (MDD) is well-documented, but the cellular and molecular basis of this pathology are poorly understood. Methods:Levels of DNA oxidation and gene expression of DNA damage repair enzymes were measured in Brodmann area 10 (BA10) and/or amygdala (uncinate fasciculus) white matter tissue from brains of MDD (n=10) and psychiatrically normal control donors (n=13). DNA oxidation was also measured in BA10 white matter of schizophrenia donors (n=10) and in prefrontal cortical white matter from control rats (n=8) and rats with repeated stress-induced anhedonia (n=8). Results:DNA oxidation in BA10 white matter was robustly elevated in MDD as compared to control donors, with a smaller elevation occurring in schizophrenia donors. DNA oxidation levels in psychiatrically affected donors that died by suicide did not significantly differ from DNA oxidation levels in psychiatrically affected donors dying by other causes (non-suicide). Gene expression levels of two base excision repair enzymes, PARP1 and OGG1, were robustly elevated in oligodendrocytes laser captured from BA10 and amygdala white matter of MDD donors, with smaller but significant elevations of these gene expressions in astrocytes. In rats, repeated stress-induced anhedonia, as measured by a reduction in sucrose preference, was associated with increased DNA oxidation in white, but not gray, matter. Conclusions:Cellular residents of brain white matter demonstrate markers of oxidative damage in MDD. Medications that interfere with oxidative damage or pathways activated by oxidative damage have potential to improve treatment for MDD

Radio Astronomy

Contains table of contents for Section 4 and reports on five research projects.National Science Foundation Grant AST 92-24191MIT Lincoln Laboratory Agreement BX-4975National Aeronautics and Space Administration/Goddard Space Flight Center Grant NAS 5-31376National Aeronautics and Space Administration/Goddard Space Flight Center Grant NAG5-10MIT Leaders for Manufacturing Progra

Absorption Coefficient (ABSCO) Tables for the Orbiting Carbon Observatories: Version 5.1

The accuracy of atmospheric trace gas retrievals depends directly on the accuracy of the molecular absorption model used within the retrieval algorithm. For remote sensing of well-mixed gases, such as carbon dioxide (CO₂), where the atmospheric variability is small compared to the background, the quality of the molecular absorption model is key. Recent updates to oxygen (O₂) absorption coefficients (ABSCO) for the 0.76 μm A-band and the water vapor (H₂O) continuum model within the 1.6 μm and 2.06 μm CO₂ bands used within the Orbiting Carbon Observatory (OCO-2 and OCO-3) algorithm are described here. Updates in the O₂ A-band involve the inclusion of new laboratory measurements within multispectrum fits to improve relative consistency between O₂ line shapes and collision-induced absorption (CIA). The H₂O continuum model has been updated to MTCKD v3.2, which has benefited from information from a range of laboratory studies relative to the model utilized in the previous ABSCO version. Impacts of these spectroscopy updates have been evaluated against ground-based atmospheric spectra from the Total Carbon Column Observing Network (TCCON) and within the framework of the OCO-2 algorithm, using OCO-2 soundings covering a range of atmospheric and surface conditions. The updated absorption coefficients (ABSCO version 5.1) are found to offer improved fitting residuals and reduced biases in retrieved surface pressure relative to the previous version (ABSCO v5.0) used within B8 and B9 of the OCO-2 retrieval algorithm and have been adopted for the OCO B10 Level 2 algorithm

Absorption Coefficient (ABSCO) Tables for the Orbiting Carbon Observatories: Version 5.1

The accuracy of atmospheric trace gas retrievals depends directly on the accuracy of the molecular absorption model used within the retrieval algorithm. For remote sensing of well-mixed gases, such as carbon dioxide (CO₂), where the atmospheric variability is small compared to the background, the quality of the molecular absorption model is key. Recent updates to oxygen (O₂) absorption coefficients (ABSCO) for the 0.76 μm A-band and the water vapor (H₂O) continuum model within the 1.6 μm and 2.06 μm CO₂ bands used within the Orbiting Carbon Observatory (OCO-2 and OCO-3) algorithm are described here. Updates in the O₂ A-band involve the inclusion of new laboratory measurements within multispectrum fits to improve relative consistency between O₂ line shapes and collision-induced absorption (CIA). The H₂O continuum model has been updated to MTCKD v3.2, which has benefited from information from a range of laboratory studies relative to the model utilized in the previous ABSCO version. Impacts of these spectroscopy updates have been evaluated against ground-based atmospheric spectra from the Total Carbon Column Observing Network (TCCON) and within the framework of the OCO-2 algorithm, using OCO-2 soundings covering a range of atmospheric and surface conditions. The updated absorption coefficients (ABSCO version 5.1) are found to offer improved fitting residuals and reduced biases in retrieved surface pressure relative to the previous version (ABSCO v5.0) used within B8 and B9 of the OCO-2 retrieval algorithm and have been adopted for the OCO B10 Level 2 algorithm

Multi-Messenger Gravitational Wave Searches with Pulsar Timing Arrays: Application to 3C66B Using the NANOGrav 11-year Data Set

When galaxies merge, the supermassive black holes in their centers may form binaries and, during the process of merger, emit low-frequency gravitational radiation in the process. In this paper we consider the galaxy 3C66B, which was used as the target of the first multi-messenger search for gravitational waves. Due to the observed periodicities present in the photometric and astrometric data of the source of the source, it has been theorized to contain a supermassive black hole binary. Its apparent 1.05-year orbital period would place the gravitational wave emission directly in the pulsar timing band. Since the first pulsar timing array study of 3C66B, revised models of the source have been published, and timing array sensitivities and techniques have improved dramatically. With these advances, we further constrain the chirp mass of the potential supermassive black hole binary in 3C66B to less than $(1.65\pm0.02) \times 10^9~{M_\odot}$ using data from the NANOGrav 11-year data set. This upper limit provides a factor of 1.6 improvement over previous limits, and a factor of 4.3 over the first search done. Nevertheless, the most recent orbital model for the source is still consistent with our limit from pulsar timing array data. In addition, we are able to quantify the improvement made by the inclusion of source properties gleaned from electromagnetic data to `blind' pulsar timing array searches. With these methods, it is apparent that it is not necessary to obtain exact a priori knowledge of the period of a binary to gain meaningful astrophysical inferences.Comment: 14 pages, 6 figures. Accepted by Ap