LPS-induced inflammatory response triggers cell cycle reactivation in murine neuronal cells through retinoblastoma proteins induction.

PubMed

D'Angelo, Barbara; Astarita, Carlo; Boffo, Silvia; Massaro-Giordano, Mina; Antonella Ianuzzi, Carmelina; Caporaso, Antonella; Macaluso, Marcella; Giordano, Antonio

2017-01-01

Cell cycle reactivation in adult neurons is an early hallmark of neurodegeneration. The lipopolysaccharide (LPS) is a well-known pro-inflammatory factor that provokes neuronal cell death via glial cells activation. The retinoblastoma (RB) family includes RB1/p105, retinoblastoma-like 1 (RBL1/p107), and retinoblastoma-like 2 (Rb2/p130). Several studies have indicated that RB proteins exhibit tumor suppressor activities, and play a central role in cell cycle regulation. In this study, we assessed LPS-mediated inflammatory effect on cell cycle reactivation and apoptosis of neuronally differentiated cells. Also, we investigated whether the LPS-mediated inflammatory response can influence the function and expression of RB proteins. Our results showed that LPS challenges triggered cell cycle reactivation of differentiated neuronal cells, indicated by an accumulation of cells in S and G2/M phase. Furthermore, we found that LPS treatment also induced apoptotic death of neurons. Interestingly, we observed that LPS-mediated inflammatory effect on cell cycle re-entry and apoptosis was concomitant with the aberrant expression of RBL1/p107 and RB1/p105. To the best of our knowledge, our study is the first to indicate a role of LPS in inducing cell cycle re-entry and/or apoptosis of differentiated neuronal cells, perhaps through mechanisms altering the expression of specific members of RB family proteins. This study provides novel information on the biology of post-mitotic neurons and could help in identifying novel therapeutic targets to prevent de novo cell cycle reactivation and/or apoptosis of neurons undergoing neurodegenerative processes.

Inducing myoblast re-entry into the cell cycle: a potential mechanism for laser-enhanced skeletal muscle regeneration

NASA Astrophysics Data System (ADS)

Liu, T.; Fang, Y.; Zhang, C. P.; Chen, P.; Wang, C. Z.; Kang, H. X.; Shen, B. J.; Liang, J.; Fu, X. B.

2014-09-01

This study investigated the effect of low-level laser irradiation (LLLI) on the cell cycle and proliferative activity of cultured myoblasts, and sought to elucidate the possible cellular mechanism by which LLLI promotes the regeneration of skeletal muscle in vivo. Primary myoblasts isolated from rat hindlegs were irradiated with helium-neon laser light at different energy densities. Distributions of cell-cycle subpopulations and the expression of cell-cycle regulatory proteins in myoblasts were assessed using flow cytometric analysis and western blot assay. It was found that laser irradiation stimulated cell-cycle entry; induced the expression of cyclin A and cyclin D; and increased cell proliferation index and bromodeoxyuridine incorporation as compared to the unirradiated control cells, indicating LLLI augmented the number of proliferative myoblasts in the S phase and G2/M phase of the cell cycle. These results suggest that LLLI at certain fluxes and wavelengths could activate quiescent myoblasts, leading to cell division and facilitating new myofiber formation. This could contribute to the improvement of skeletal muscle regeneration following trauma and myopathic diseases.

Cell-cycle quiescence maintains Caenorhabditis elegans germline stem cells independent of GLP-1/Notch

PubMed Central

Seidel, Hannah S; Kimble, Judith

2015-01-01

Many types of adult stem cells exist in a state of cell-cycle quiescence, yet it has remained unclear whether quiescence plays a role in maintaining the stem cell fate. Here we establish the adult germline of Caenorhabditis elegans as a model for facultative stem cell quiescence. We find that mitotically dividing germ cells—including germline stem cells—become quiescent in the absence of food. This quiescence is characterized by a slowing of S phase, a block to M-phase entry, and the ability to re-enter M phase rapidly in response to re-feeding. Further, we demonstrate that cell-cycle quiescence alters the genetic requirements for stem cell maintenance: The signaling pathway required for stem cell maintenance under fed conditions—GLP-1/Notch signaling—becomes dispensable under conditions of quiescence. Thus, cell-cycle quiescence can itself maintain stem cells, independent of the signaling pathway otherwise essential for such maintenance. DOI: http://dx.doi.org/10.7554/eLife.10832.001 PMID:26551561

Restrictions in Cell Cycle Progression of Adult Vestibular Supporting Cells in Response to Ectopic Cyclin D1 Expression

PubMed Central

Loponen, Heidi; Ylikoski, Jukka; Albrecht, Jeffrey H.; Pirvola, Ulla

2011-01-01

Sensory hair cells and supporting cells of the mammalian inner ear are quiescent cells, which do not regenerate. In contrast, non-mammalian supporting cells have the ability to re-enter the cell cycle and produce replacement hair cells. Earlier studies have demonstrated cyclin D1 expression in the developing mouse supporting cells and its downregulation along maturation. In explant cultures of the mouse utricle, we have here focused on the cell cycle control mechanisms and proliferative potential of adult supporting cells. These cells were forced into the cell cycle through adenoviral-mediated cyclin D1 overexpression. Ectopic cyclin D1 triggered robust cell cycle re-entry of supporting cells, accompanied by changes in p27Kip1 and p21Cip1 expressions. Main part of cell cycle reactivated supporting cells were DNA damaged and arrested at the G2/M boundary. Only small numbers of mitotic supporting cells and rare cells with signs of two successive replications were found. Ectopic cyclin D1-triggered cell cycle reactivation did not lead to hyperplasia of the sensory epithelium. In addition, a part of ectopic cyclin D1 was sequestered in the cytoplasm, reflecting its ineffective nuclear import. Combined, our data reveal intrinsic barriers that limit proliferative capacity of utricular supporting cells. PMID:22073316

Restrictions in cell cycle progression of adult vestibular supporting cells in response to ectopic cyclin D1 expression.

PubMed

Loponen, Heidi; Ylikoski, Jukka; Albrecht, Jeffrey H; Pirvola, Ulla

2011-01-01

Sensory hair cells and supporting cells of the mammalian inner ear are quiescent cells, which do not regenerate. In contrast, non-mammalian supporting cells have the ability to re-enter the cell cycle and produce replacement hair cells. Earlier studies have demonstrated cyclin D1 expression in the developing mouse supporting cells and its downregulation along maturation. In explant cultures of the mouse utricle, we have here focused on the cell cycle control mechanisms and proliferative potential of adult supporting cells. These cells were forced into the cell cycle through adenoviral-mediated cyclin D1 overexpression. Ectopic cyclin D1 triggered robust cell cycle re-entry of supporting cells, accompanied by changes in p27(Kip1) and p21(Cip1) expressions. Main part of cell cycle reactivated supporting cells were DNA damaged and arrested at the G2/M boundary. Only small numbers of mitotic supporting cells and rare cells with signs of two successive replications were found. Ectopic cyclin D1-triggered cell cycle reactivation did not lead to hyperplasia of the sensory epithelium. In addition, a part of ectopic cyclin D1 was sequestered in the cytoplasm, reflecting its ineffective nuclear import. Combined, our data reveal intrinsic barriers that limit proliferative capacity of utricular supporting cells.

Fad24, a Positive Regulator of Adipogenesis, Is Required for S Phase Re-entry of C2C12 Myoblasts Arrested in G0 Phase and Involved in p27(Kip1) Expression at the Protein Level.

PubMed

Ochiai, Natsuki; Nishizuka, Makoto; Osada, Shigehiro; Imagawa, Masayoshi

2016-05-01

Factor for adipocyte differentiation 24 (fad24) is a positive regulator of adipogenesis. We previously found that human fad24 is abundantly expressed in skeletal muscle. However, the function of fad24 in skeletal muscle remains largely unknown. Because skeletal muscle is a highly regenerative tissue, we focused on the function of fad24 in skeletal muscle regeneration. In this paper, we investigated the role of fad24 in the cell cycle re-entry of quiescent C2C12 myoblasts-mimicked satellite cells. The expression levels of fad24 and histone acetyltransferase binding to ORC1 (hbo1), a FAD24-interacting factor, were elevated at the early phase of the regeneration process in response to cardiotoxin-induced muscle injury. The knockdown of fad24 inhibited the proliferation of quiescent myoblasts, whereas fad24 knockdown did not affect differentiation. S phase entry following serum activation is abrogated by fad24 knockdown in quiescent cells. Furthermore, fad24 knockdown cells show a marked accumulation of p27(Kip1) protein. These results suggest that fad24 may have an important role in the S phase re-entry of quiescent C2C12 cells through the regulation of p27(Kip1) at the protein level.

Characterisation of re-entrant circuit (or rotational activity) in vitro using the HL1-6 myocyte cell line.

PubMed

Houston, Charles; Tzortzis, Konstantinos N; Roney, Caroline; Saglietto, Andrea; Pitcher, David S; Cantwell, Chris D; Chowdhury, Rasheda A; Ng, Fu Siong; Peters, Nicholas S; Dupont, Emmanuel

2018-06-01

Fibrillation is the most common arrhythmia observed in clinical practice. Understanding of the mechanisms underlying its initiation and maintenance remains incomplete. Functional re-entries are potential drivers of the arrhythmia. Two main concepts are still debated, the "leading circle" and the "spiral wave or rotor" theories. The homogeneous subclone of the HL1 atrial-derived cardiomyocyte cell line, HL1-6, spontaneously exhibits re-entry on a microscopic scale due to its slow conduction velocity and the presence of triggers, making it possible to examine re-entry at the cellular level. We therefore investigated the re-entry cores in cell monolayers through the use of fluorescence optical mapping at high spatiotemporal resolution in order to obtain insights into the mechanisms of re-entry. Re-entries in HL1-6 myocytes required at least two triggers and a minimum colony area to initiate (3.5 to 6.4 mm 2 ). After electrical activity was completely stopped and re-started by varying the extracellular K + concentration, re-entries never returned to the same location while 35% of triggers re-appeared at the same position. A conduction delay algorithm also allows visualisation of the core of the re-entries. This work has revealed that the core of re-entries is conduction blocks constituted by lines and/or groups of cells rather than the round area assumed by the other concepts of functional re-entry. This highlights the importance of experimentation at the microscopic level in the study of re-entry mechanisms. Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.

The endocrine dyscrasia that accompanies menopause and andropause induces aberrant cell cycle signaling that triggers re-entry of post-mitotic neurons into the cell cycle, neurodysfunction, neurodegeneration and cognitive disease.

PubMed

Atwood, Craig S; Bowen, Richard L

2015-11-01

This article is part of a Special Issue "SBN 2014". Sex hormones are physiological factors that promote neurogenesis during embryonic and fetal development. During childhood and adulthood these hormones support the maintenance of brain structure and function via neurogenesis and the formation of dendritic spines, axons and synapses required for the capture, processing and retrieval of information (memories). Not surprisingly, changes in these reproductive hormones that occur with menopause and during andropause are strongly correlated with neurodegeneration and cognitive decline. In this connection, much evidence now indicates that Alzheimer's disease (AD) involves aberrant re-entry of post-mitotic neurons into the cell cycle. Cell cycle abnormalities appear very early in the disease, prior to the appearance of plaques and tangles, and explain the biochemical, neuropathological and cognitive changes observed with disease progression. Intriguingly, a recent animal study has demonstrated that induction of adult neurogenesis results in the loss of previously encoded memories while decreasing neurogenesis after memory formation during infancy mitigated forgetting. Here we review the biochemical, epidemiological and clinical evidence that alterations in sex hormone signaling associated with menopause and andropause drive the aberrant re-entry of post-mitotic neurons into an abortive cell cycle that leads to neurite retraction, neuron dysfunction and neuron death. When the reproductive axis is in balance, gonadotropins such as luteinizing hormone (LH), and its fetal homolog, human chorionic gonadotropin (hCG), promote pluripotent human and totipotent murine embryonic stem cell and neuron proliferation. However, strong evidence supports menopausal/andropausal elevations in the LH:sex steroid ratio as driving aberrant mitotic events. These include the upregulation of tumor necrosis factor; amyloid-β precursor protein processing towards the production of mitogenic Aβ; and the activation of Cdk5, a key regulator of cell cycle progression and tau phosphorylation (a cardinal feature of both neurogenesis and neurodegeneration). Cognitive and biochemical studies confirm the negative consequences of a high LH:sex steroid ratio on dendritic spine density and human cognitive performance. Prospective epidemiological and clinical evidence in humans supports the premise that rebalancing the ratio of circulating gonadotropins:sex steroids reduces the incidence of AD. Together, these data support endocrine dyscrasia and the subsequent loss of cell cycle control as an important etiological event in the development of neurodegenerative diseases including AD, stroke and Parkinson's disease. Published by Elsevier Inc.

HSV-1 infection of human corneal epithelial cells: receptor-mediated entry and trends of re-infection.

PubMed

Shah, Arpeet; Farooq, Asim V; Tiwari, Vaibhav; Kim, Min-Jung; Shukla, Deepak

2010-11-20

The human cornea is a primary target for herpes simplex virus-1 (HSV-1) infection. The goals of the study were to determine the cellular modalities of HSV-1 entry into human corneal epithelial (HCE) cells. Specific features of the study included identifying major entry receptors, assessing pH dependency, and determining trends of re-infection. A recombinant HSV-1 virus expressing beta-galactosidase was used to ascertain HSV-1 entry into HCE cells. Viral replication within cells was confirmed using a time point plaque assay. Lysosomotropic agents were used to test for pH dependency of entry. Flow cytometry and immunocytochemistry were used to determine expression of three cellular receptors--nectin-1, herpesvirus entry mediator (HVEM), and paired immunoglobulin-like 2 receptor alpha (PILR-a). The necessity of these receptors for viral entry was tested using antibody-blocking. Finally, trends of re-infection were investigated using viral entry assay and flow cytometry post-primary infection. Cultured HCE cells showed high susceptibility to HSV-1 entry and replication. Entry was demonstrated to be pH dependent as blocking vesicular acidification decreased entry. Entry receptors expressed on the cell membrane include nectin-1, HVEM, and PILR-α. Receptor-specific antibodies blocked entry receptors, reduced viral entry and indicated nectin-1 as the primary receptor used for entry. Cells re-infected with HSV-1 showed a decrease in entry, which was correlated to decreased levels of nectin-1 as demonstrated by flow cytometry. HSV-1 is capable of developing an infection in HCE cells using a pH dependent entry process that involves primarily nectin-1 but also the HVEM and PILR-α receptors. Re-infected cells show decreased levels of entry, correlated with a decreased level of nectin-1 receptor expression.

Toll-like receptor 4 is involved in the cell cycle modulation and required for effective human cytomegalovirus infection in THP-1 macrophages

DOE Office of Scientific and Technical Information (OSTI.GOV)

Arcangeletti, Maria-Cristina, E-mail: mariacristina.arcangeletti@unipr.it; Germini, Diego; Rodighiero, Isabella

2013-05-25

Suitable host cell metabolic conditions are fundamental for the effective development of the human cytomegalovirus (HCMV) lytic cycle. Indeed, several studies have demonstrated the ability of this virus to interfere with cell cycle regulation, mainly by blocking proliferating cells in G1 or G1/S. In the present study, we demonstrate that HCMV deregulates the cell cycle of THP-1 macrophages (a cell line irreversibly arrested in G0) by pushing them into S and G2 phases. Moreover, we show that HCMV infection of THP-1 macrophages leads to Toll-like receptor 4 (TLR4) activation. Since various studies have indicated TLR4 to be involved in promotingmore » cell proliferation, here we investigate the possible role of TLR4 in the observed HCMV-induced cell cycle perturbation. Our data strongly support TLR4 as a mediator of HCMV-triggered cell cycle activation in THP-1 macrophages favouring, in turn, the development of an efficient viral lytic cycle. - Highlights: ► We studied HCMV infection impact on THP-1 macrophage cell cycle. ► We analysed the role played by Toll-like receptor (TLR) 4 upon HCMV infection. ► HCMV pushes THP-1 macrophages (i.e. resting cells) to re-enter the cell cycle. ► TLR4 pathway inhibition strongly affects the effectiveness of HCMV replication. ► TLR4 pathway inhibition significantly decreases HCMV-induced cell cycle re-entry.« less

A Study on Re-entry Predictions of Uncontrolled Space Objects for Space Situational Awareness

NASA Astrophysics Data System (ADS)

Choi, Eun-Jung; Cho, Sungki; Lee, Deok-Jin; Kim, Siwoo; Jo, Jung Hyun

2017-12-01

The key risk analysis technologies for the re-entry of space objects into Earth’s atmosphere are divided into four categories: cataloguing and databases of the re-entry of space objects, lifetime and re-entry trajectory predictions, break-up models after re-entry and multiple debris distribution predictions, and ground impact probability models. In this study, we focused on re- entry prediction, including orbital lifetime assessments, for space situational awareness systems. Re-entry predictions are very difficult and are affected by various sources of uncertainty. In particular, during uncontrolled re-entry, large spacecraft may break into several pieces of debris, and the surviving fragments can be a significant hazard for persons and properties on the ground. In recent years, specific methods and procedures have been developed to provide clear information for predicting and analyzing the re-entry of space objects and for ground-risk assessments. Representative tools include object reentry survival analysis tool (ORSAT) and debris assessment software (DAS) developed by National Aeronautics and Space Administration (NASA), spacecraft atmospheric re-entry and aerothermal break-up (SCARAB) and debris risk assessment and mitigation analysis (DRAMA) developed by European Space Agency (ESA), and semi-analytic tool for end of life analysis (STELA) developed by Centre National d’Etudes Spatiales (CNES). In this study, various surveys of existing re-entry space objects are reviewed, and an efficient re-entry prediction technique is suggested based on STELA, the life-cycle analysis tool for satellites, and DRAMA, a re-entry analysis tool. To verify the proposed method, the re-entry of the Tiangong-1 Space Lab, which is expected to re-enter Earth’s atmosphere shortly, was simulated. Eventually, these results will provide a basis for space situational awareness risk analyses of the re-entry of space objects.

Circumferential and functional re-entry of in vivo slow-wave activity in the porcine small intestine.

PubMed

Angeli, T R; O'Grady, G; Du, P; Paskaranandavadivel, N; Pullan, A J; Bissett, I P; Cheng, L K

2013-05-01

Slow-waves modulate the pattern of small intestine contractions. However, the large-scale spatial organization of intestinal slow-wave pacesetting remains uncertain because most previous studies have had limited resolution. This study applied high-resolution (HR) mapping to evaluate intestinal pacesetting mechanisms and propagation patterns in vivo. HR serosal mapping was performed in anesthetized pigs using flexible arrays (256 electrodes; 32 × 8; 4 mm spacing), applied along the jejunum. Slow-wave propagation patterns, frequencies, and velocities were calculated. Slow-wave initiation sources were identified and analyzed by animation and isochronal activation mapping. Analysis comprised 32 recordings from nine pigs (mean duration 5.1 ± 3.9 min). Slow-wave propagation was analyzed, and a total of 26 sources of slow-wave initiation were observed and classified as focal pacemakers (31%), sites of functional re-entry (23%) and circumferential re-entry (35%), or indeterminate sources (11%). The mean frequencies of circumferential and functional re-entry were similar (17.0 ± 0.3 vs 17.2 ± 0.4 cycle min(-1) ; P = 0.5), and greater than that of focal pacemakers (12.7 ± 0.8 cycle min(-1) ; P < 0.001). Velocity was anisotropic (12.9 ± 0.7 mm s(-1) circumferential vs 9.0 ± 0.7 mm s(-1) longitudinal; P < 0.05), contributing to the onset and maintenance of re-entry. This study has shown multiple patterns of slow-wave initiation in the jejunum of anesthetized pigs. These results constitute the first description and analysis of circumferential re-entry in the gastrointestinal tract and functional re-entry in the in vivo small intestine. Re-entry can control the direction, pattern, and frequency of slow-wave propagation, and its occurrence and functional significance merit further investigation. © 2013 Blackwell Publishing Ltd.

mTOR and Neuronal Cell Cycle Re-entry: How Impaired Brain Insulin Signaling Promotes Alzheimer's Disease

PubMed Central

Norambuena, Andrés; Wallrabe, Horst; McMahon, Lloyd; Silva, Antonia; Swanson, Eric; Khan, Shahzad S.; Baerthlein, Daniel; Kodis, Erin; Oddo, Salvatore; Mandell, James W.; Bloom, George S.

2016-01-01

A major obstacle to pre-symptomatic diagnosis and disease-modifying therapy for Alzheimer's disease (AD) is inadequate understanding of molecular mechanisms of AD pathogenesis. For example, impaired brain insulin signaling is an AD hallmark, but whether and how it might contribute to the synaptic dysfunction and neuron death that underlie memory and cognitive impairment has been mysterious. Neuron death in AD is often caused by cell cycle re-entry (CCR) mediated by amyloid-β oligomers (AβOs) and tau, the precursors of plaques and tangles. We now report that CCR results from AβO-induced activation of the protein kinase complex, mTORC1, at the plasma membrane and mTORC1-dependent tau phosphorylation, and that CCR can be prevented by insulin-stimulated activation of lysosomal mTORC1. AβOs were also shown previously to reduce neuronal insulin signaling. Our data therefore indicate that the decreased insulin signaling provoked by AβOs unleashes their toxic potential to cause neuronal CCR, and by extension, neuron death. PMID:27693185

Trichinella spiralis: nurse cell formation with emphasis on analogy to muscle cell repair

PubMed Central

Wu, Zhiliang; Sofronic-Milosavljevic, Lj; Nagano, Isao; Takahashi, Yuzo

2008-01-01

Trichinella infection results in formation of a capsule in infected muscles. The capsule is a residence of the parasite which is composed of the nurse cell and fibrous wall. The process of nurse cell formation is complex and includes infected muscle cell response (de-differentiation, cell cycle re-entry and arrest) and satellite cell responses (activation, proliferation and differentiation). Some events that occur during the nurse cell formation are analogous to those occurring during muscle cell regeneration/repair. This article reviews capsule formation with emphasis on this analogy. PMID:18710582

Metformin inhibits cell cycle progression of B-cell chronic lymphocytic leukemia cells.

PubMed

Bruno, Silvia; Ledda, Bernardetta; Tenca, Claudya; Ravera, Silvia; Orengo, Anna Maria; Mazzarello, Andrea Nicola; Pesenti, Elisa; Casciaro, Salvatore; Racchi, Omar; Ghiotto, Fabio; Marini, Cecilia; Sambuceti, Gianmario; DeCensi, Andrea; Fais, Franco

2015-09-08

B-cell chronic lymphocytic leukemia (CLL) was believed to result from clonal accumulation of resting apoptosis-resistant malignant B lymphocytes. However, it became increasingly clear that CLL cells undergo, during their life, iterative cycles of re-activation and subsequent clonal expansion. Drugs interfering with CLL cell cycle entry would be greatly beneficial in the treatment of this disease. 1, 1-Dimethylbiguanide hydrochloride (metformin), the most widely prescribed oral hypoglycemic agent, inexpensive and well tolerated, has recently received increased attention for its potential antitumor activity. We wondered whether metformin has apoptotic and anti-proliferative activity on leukemic cells derived from CLL patients. Metformin was administered in vitro either to quiescent cells or during CLL cell activation stimuli, provided by classical co-culturing with CD40L-expressing fibroblasts. At doses that were totally ineffective on normal lymphocytes, metformin induced apoptosis of quiescent CLL cells and inhibition of cell cycle entry when CLL were stimulated by CD40-CD40L ligation. This cytostatic effect was accompanied by decreased expression of survival- and proliferation-associated proteins, inhibition of signaling pathways involved in CLL disease progression and decreased intracellular glucose available for glycolysis. In drug combination experiments, metformin lowered the apoptotic threshold and potentiated the cytotoxic effects of classical and novel antitumor molecules. Our results indicate that, while CLL cells after stimulation are in the process of building their full survival and cycling armamentarium, the presence of metformin affects this process.

Vitamin A-Retinoic Acid Signaling Regulates Hematopoietic Stem Cell Dormancy.

PubMed

Cabezas-Wallscheid, Nina; Buettner, Florian; Sommerkamp, Pia; Klimmeck, Daniel; Ladel, Luisa; Thalheimer, Frederic B; Pastor-Flores, Daniel; Roma, Leticia P; Renders, Simon; Zeisberger, Petra; Przybylla, Adriana; Schönberger, Katharina; Scognamiglio, Roberta; Altamura, Sandro; Florian, Carolina M; Fawaz, Malak; Vonficht, Dominik; Tesio, Melania; Collier, Paul; Pavlinic, Dinko; Geiger, Hartmut; Schroeder, Timm; Benes, Vladimir; Dick, Tobias P; Rieger, Michael A; Stegle, Oliver; Trumpp, Andreas

2017-05-18

Dormant hematopoietic stem cells (dHSCs) are atop the hematopoietic hierarchy. The molecular identity of dHSCs and the mechanisms regulating their maintenance or exit from dormancy remain uncertain. Here, we use single-cell RNA sequencing (RNA-seq) analysis to show that the transition from dormancy toward cell-cycle entry is a continuous developmental path associated with upregulation of biosynthetic processes rather than a stepwise progression. In addition, low Myc levels and high expression of a retinoic acid program are characteristic for dHSCs. To follow the behavior of dHSCs in situ, a Gprc5c-controlled reporter mouse was established. Treatment with all-trans retinoic acid antagonizes stress-induced activation of dHSCs by restricting protein translation and levels of reactive oxygen species (ROS) and Myc. Mice maintained on a vitamin A-free diet lose HSCs and show a disrupted re-entry into dormancy after exposure to inflammatory stress stimuli. Our results highlight the impact of dietary vitamin A on the regulation of cell-cycle-mediated stem cell plasticity. VIDEO ABSTRACT. Copyright © 2017. Published by Elsevier Inc.

Early induction of c-Myc is associated with neuronal cell death.

PubMed

Lee, Hyun-Pil; Kudo, Wataru; Zhu, Xiongwei; Smith, Mark A; Lee, Hyoung-gon

2011-11-14

Neuronal cell cycle activation has been implicated in neurodegenerative diseases such as Alzheimer's disease, while the initiating mechanism of cell cycle activation remains to be determined. Interestingly, our previous studies have shown that cell cycle activation by c-Myc (Myc) leads to neuronal cell death which suggests Myc might be a key regulator of cell cycle re-entry mediated neuronal cell death. However, the pattern of Myc expression in the process of neuronal cell death has not been addressed. To this end, we examined Myc induction by the neurotoxic agents camptothecin and amyloid-β peptide in a differentiated SH-SY5Y neuronal cell culture model. Myc expression was found to be significantly increased following either treatment and importantly, the induction of Myc preceded neuronal cell death suggesting it is an early event of neuronal cell death. Since ectopic expression of Myc in neurons causes the cell cycle activation and neurodegeneration in vivo, the current data suggest that induction of Myc by neurotoxic agents or other disease factors might be a key mediator in cell cycle activation and consequent cell death that is a feature of neurodegenerative diseases. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

Re-entry and reintegration: returning home after combat.

PubMed

Doyle, Michael E; Peterson, Kris A

2005-01-01

Soldier life exists on a continuum of readiness for deployment. Re-entry and reintegration-the return home and reunion with family and community-key the success of the deployment cycle. In current and projected future operations, the Army and society will both bear the burden of this re-entry and re-integration. Programs and procedures in place work towards improving communication, mitigating distress and resolving crises during reentry and reintegration. Key elements include: inclusion of families and communities early into the planning for reentry and reintegration; normalization (non-medicalization of distress); easy access to behavioral health professionals; and education of families on resources and benefits. Through broad collaboration, maximal benefit to the Soldier, family members and society be realized.

The Endocrine Dyscrasia that Accompanies Menopause and Andropause Induces Aberrant Cell Cycle Signaling that Triggers Cell Cycle Reentry of Post-mitotic Neurons, Neurodysfunction, Neurodegeneration and Cognitive Disease

PubMed Central

Atwood, Craig S.; Bowen, Richard L.

2016-01-01

Sex hormones are the physiological factors that regulate neurogenesis during embryogenesis and continuing through adulthood. These hormones support the formation of brain structures such as dendritic spines, axons and synapses required for the capture of information (memories). Intriguingly, a recent animal study has demonstrated that induction of neurogenesis results in the loss of previously encoded memories in animals (e.g. infantile amnesia). In this connection, much evidence now indicates that Alzheimer’s disease (AD) also involves aberrant re-entry of post-mitotic neurons into the cell cycle. Cell cycle abnormalities appear very early in the disease, prior to the appearance of plaques and tangles, and explain the biochemical, neuropathological and cognitive changes observed with disease progression. Since sex hormones control when and how neurons proliferate and differentiate, the endocrine dyscrasia that accompanies menopause and andropause is a key signaling event that impacts neurogenesis and the acquisition, processing, storage and recall of memories. Here we review the biochemical, epidemiological and clinical evidence that alterations in endocrine signaling with menopause and andropause drive the aberrant re-entry of post-mitotic neurons into an abortive cell cycle with neurite retraction that leads to neuron dysfunction and death. When the reproductive axis is in balance, luteinizing hormone (LH), and its fetal homolog, human chorionic gonadotropin (hCG), promote pluripotent human and totipotent murine embryonic stem cell and neuron proliferation. However, strong evidence supports menopausal/andropausal elevations in the ratio of LH:sex steroids as driving aberrant mitotic events mediated by the upregulation of tumor necrosis factor, amyloid-β precursor protein processing towards the production of mitogenic Aβ, and the activation of Cdk5, a key regulator of cell cycle progression and tau phosphorylation (a cardinal feature of both neurogenesis and neurodegeneration). Cognitive studies also demonstrate the negative consequences of a high LH:sex steroid ratio on human cognitive performance. Prospective epidemiological and clinical evidence in humans supports lowering the ratio of circulating gonadotropins-GnRH to sex steroids in reducing the incidence of AD and halting cognitive decline. Together, these data support endocrine dyscrasia and the subsequent loss of cell cycle control as an important etiological event in the development of neurodegenerative diseases including AD, stroke and Parkinson’s disease. PMID:26188949

Behavior of HfB2-SiC Materials in Simulated Re-Entry Environments

NASA Technical Reports Server (NTRS)

Ellerby, Don; Beckman, Sarah; Irby, Edward; Johnson, Sylvia M.; Gunsman, Michael; Gasch, Matthew; Ridge, Jerry; Martinez, Ed; Squire, Tom; Olejniczak, Joe

2003-01-01

The objectives of this research are to: 1) Investigate the oxidation/ablation behavior of HfB2/SiC materials in simulated re-entry environments; 2) Use the arc jet test results to define appropriate use environments for these materials for use in vehicle design. The parameters to be investigated include: surface temperature, stagnation pressure, duration, number of cycles, and thermal stresses.

Cell cycle activation in p21 dependent pathway: An alternative mechanism of organophosphate induced dopaminergic neurodegeneration.

PubMed

Wani, Willayat Yousuf; Kandimalla, Ramesh J L; Sharma, Deep Raj; Kaushal, Alka; Ruban, Anand; Sunkaria, Aditya; Vallamkondu, Jayalakshmi; Chiarugi, Alberto; Reddy, P Hemachandra; Gill, Kiran Dip

2017-07-01

In the previous study, we demonstrated that dichlorvos induces oxidative stress in dopaminergic neuronal cells and subsequent caspase activation mediates apoptosis. In the present study, we evaluated the effect and mechanism of dichlorvos induced oxidative stress on cell cycle activation in NGF-differentiated PC12 cells. Dichlorvos exposure resulted in oxidative DNA damage along with activation of cell cycle machinery in differentiated PC12 cells. Dichlorvos exposed cells exhibited an increased expression of p53, cyclin-D1, pRb and decreased expression of p21suggesting a re-entry of differentiated cells into the cell cycle. Cell cycle analysis of dichlorvos exposed cells revealed a reduction of cells in the G 0 /G 1 phase of the cell cycle (25%), and a concomitant increase of cells in S phase (30%) and G2/M phase (43.3%) compared to control PC12 cells. Further, immunoblotting of cytochrome c, Bax, Bcl-2 and cleaved caspase-3 revealed that dichlorvos induces a caspase-dependent cell death in PC12 cells. These results suggest that Dichlorvos exposure has the potential to generate oxidative stress which evokes activation of cell cycle machinery leading to apoptotic cell death via cytochrome c release from mitochondria and subsequent caspase-3 activation in differentiated PC12 cells. Copyright © 2016 Elsevier B.V. All rights reserved.

Phosphorylation of Smad2/3 at the specific linker threonine residue indicates slow-cycling esophageal stem-like cells before re-entry to the cell cycle.

PubMed

Takahashi, Y; Fukui, T; Kishimoto, M; Suzuki, R; Mitsuyama, T; Sumimoto, K; Okazaki, T; Sakao, M; Sakaguchi, Y; Yoshida, K; Uchida, K; Nishio, A; Matsuzaki, K; Okazaki, K

2016-01-01

The stem cell compartment in the esophageal epithelium is possibly located in the basal layer. We have identified significant expression of Smad2/3, phosphorylated at specific linker threonine residues (pSmad2/3L-Thr), in the epithelial cells of murine stomach and intestine, and have suggested that these cells are epithelial stem cells. In this study, we explore whether pSmad2/3L-Thr could serve as a biomarker for esophageal stem cells. We examined esophageal tissues from normal C57BL/6 mice and those with esophagitis. Double immunofluorescent staining of pSmad2/3L-Thr with Ki67, CDK4, p63, or CK14 was performed. After immunofluorescent staining, we stained the same sections with hematoxylin-eosin and observed these cells under a light microscope. We used the 5-bromo-2-deoxyuridine (BrdU) labeling assay to examine label retention of pSmad2/3L-Thr immunostaining-positive cells. We collected specimens 5, 10, 15 and 20 days after repeated BrdU administrations and observed double immunofluorescent staining of pSmad2/3L-Thr with BrdU. In the esophagus, pSmad2/3L-Thr immunostaining-positive cells were detected in the basal layer. These cells were detected between Ki67 immunostaining-positive cells, but they were not co-localized with Ki67. pSmad2/3L-Thr immunostaining-positive cells showed co-localization with CDK4, p63, and CK14. Under a light microscope, pSmad2/3L-Thr immunostaining-positive cells indicated undifferentiated morphological features. Until 20 days follow-up period, pSmad2/3L-Thr immunostaining-positive cells were co-localized with BrdU. pSmad2/3L-Thr immunostaining-positive cells significantly increased in the regeneration phase of esophagitis mucosae, as compared with control mice (esophagitis vs. 6.889 ± 0.676/cm vs. 4.293 ± 0.659/cm; P < 0.001). We have identified significant expression of pSmad2/3L-Thr in the specific epithelial cells of murine esophagi. We suggest that these cells are slow-cycling epithelial stem-like cells before re-entry to the cell cycle. © 2016 International Society for Diseases of the Esophagus.

Increased Re-Entry into Cell Cycle Mitigates Age-Related Neurogenic Decline in the Murine Subventricular Zone

PubMed Central

Stoll, Elizabeth A.; Habibi, Behnum A.; Mikheev, Andrei M.; Lasiene, Jurate; Massey, Susan C.; Swanson, Kristin R.; Rostomily, Robert C.; Horner, Philip J.

2012-01-01

Although new neurons are produced in the subventricular zone (SVZ) of the adult mammalian brain, fewer functional neurons are produced with increasing age. The age-related decline in neurogenesis has been attributed to a decreased pool of neural progenitor cells (NPCs), an increased rate of cell death, and an inability to undergo neuronal differentiation and develop functional synapses. The time between mitotic events has also been hypothesized to increase with age, but this has not been directly investigated. Studying primary-cultured NPCs from the young adult and aged mouse forebrain, we observe that fewer aged cells are dividing at a given time; however, the mitotic cells in aged cultures divide more frequently than mitotic cells in young cultures during a 48-hour period of live-cell time-lapse imaging. Double-thymidine-analog labeling also demonstrates that fewer aged cells are dividing at a given time, but those that do divide are significantly more likely to re-enter the cell cycle within a day, both in vitro and in vivo. Meanwhile, we observed that cellular survival is impaired in aged cultures. Using our live-cell imaging data, we developed a mathematical model describing cell cycle kinetics to predict the growth curves of cells over time in vitro and the labeling index over time in vivo. Together, these data surprisingly suggest that progenitor cells remaining in the aged SVZ are highly proliferative. PMID:21948688

A dual transcriptional reporter and CDK-activity sensor marks cell cycle entry and progression in C. elegans

PubMed Central

van Rijnberk, Lotte M.; van der Horst, Suzanne E. M.; van den Heuvel, Sander; Ruijtenberg, Suzan

2017-01-01

Development, tissue homeostasis and tumor suppression depend critically on the correct regulation of cell division. Central in the cell division process is the decision whether to enter the next cell cycle and commit to going through the S and M phases, or to remain temporarily or permanently arrested. Cell cycle studies in genetic model systems could greatly benefit from visualizing cell cycle commitment in individual cells without the need of fixation. Here, we report the development and characterization of a reporter to monitor cell cycle entry in the nematode C. elegans. This reporter combines the mcm-4 promoter, to reveal Rb/E2F-mediated transcriptional control, and a live-cell sensor for CDK-activity. The CDK sensor was recently developed for use in human cells and consists of a DNA Helicase fragment fused to eGFP. Upon phosphorylation by CDKs, this fusion protein changes in localization from the nucleus to the cytoplasm. The combined regulation of transcription and subcellular localization enabled us to visualize the moment of cell cycle entry in dividing seam cells during C. elegans larval development. This reporter is the first to reflect cell cycle commitment in C. elegans and will help further genetic studies of the mechanisms that underlie cell cycle entry and exit. PMID:28158315

Resetting the epigenome for heart regeneration.

PubMed

Quaife-Ryan, Gregory A; Sim, Choon Boon; Porrello, Enzo R; Hudson, James E

2016-10-01

In contrast to adults, recent evidence suggests that neonatal mice are able to regenerate following cardiac injury. This regenerative capacity is reliant on robust induction of cardiomyocyte proliferation, which is required for faithful regeneration of the heart following injury. However, cardiac regenerative potential is lost as cardiomyocytes mature and permanently withdraw from the cell cycle shortly after birth. Recently, a handful of factors responsible for the regenerative disparity between the adult and neonatal heart have been identified, but the proliferative response of adult cardiomyocytes following modulation of these factors rarely reaches neonatal levels. The inefficient re-induction of proliferation in adult cardiomyocytes may be due to the epigenetic landscape, which drastically changes during cardiac development and maturation. In this review, we provide an overview of the role of epigenetic modifiers in developmental processes related to cardiac regeneration. We propose an epigenetic framework for heart regeneration whereby adult cardiomyocyte identity requires resetting to a neonatal-like state to facilitate cell cycle re-entry and regeneration following cardiac injury. Copyright © 2016 Elsevier Ltd. All rights reserved.

Hedgehog regulates Norrie disease protein to drive neural progenitor self-renewal.

PubMed

McNeill, Brian; Mazerolle, Chantal; Bassett, Erin A; Mears, Alan J; Ringuette, Randy; Lagali, Pamela; Picketts, David J; Paes, Kim; Rice, Dennis; Wallace, Valerie A

2013-03-01

Norrie disease (ND) is a congenital disorder characterized by retinal hypovascularization and cognitive delay. ND has been linked to mutations in 'Norrie Disease Protein' (Ndp), which encodes the secreted protein Norrin. Norrin functions as a secreted angiogenic factor, although its role in neural development has not been assessed. Here, we show that Ndp expression is initiated in retinal progenitors in response to Hedgehog (Hh) signaling, which induces Gli2 binding to the Ndp promoter. Using a combination of genetic epistasis and acute RNAi-knockdown approaches, we show that Ndp is required downstream of Hh activation to induce retinal progenitor proliferation in the retina. Strikingly, Ndp regulates the rate of cell-cycle re-entry and not cell-cycle kinetics, thereby uncoupling the self-renewal and cell-cycle progression functions of Hh. Taken together, we have uncovered a cell autonomous function for Ndp in retinal progenitor proliferation that is independent of its function in the retinal vasculature, which could explain the neural defects associated with ND.

Recovery from the DNA Replication Checkpoint

PubMed Central

Chaudhury, Indrajit; Koepp, Deanna M.

2016-01-01

Checkpoint recovery is integral to a successful checkpoint response. Checkpoint pathways monitor progress during cell division so that in the event of an error, the checkpoint is activated to block the cell cycle and activate repair pathways. Intrinsic to this process is that once repair has been achieved, the checkpoint signaling pathway is inactivated and cell cycle progression resumes. We use the term “checkpoint recovery” to describe the pathways responsible for the inactivation of checkpoint signaling and cell cycle re-entry after the initial stress has been alleviated. The DNA replication or S-phase checkpoint monitors the integrity of DNA synthesis. When replication stress is encountered, replication forks are stalled, and the checkpoint signaling pathway is activated. Central to recovery from the S-phase checkpoint is the restart of stalled replication forks. If checkpoint recovery fails, stalled forks may become unstable and lead to DNA breaks or unusual DNA structures that are difficult to resolve, causing genomic instability. Alternatively, if cell cycle resumption mechanisms become uncoupled from checkpoint inactivation, cells with under-replicated DNA might proceed through the cell cycle, also diminishing genomic stability. In this review, we discuss the molecular mechanisms that contribute to inactivation of the S-phase checkpoint signaling pathway and the restart of replication forks during recovery from replication stress. PMID:27801838

Acute amiodarone promotes drift and early termination of spiral wave re-entry.

PubMed

Nakagawa, Harumichi; Honjo, Haruo; Ishiguro, Yuko S; Yamazaki, Masatoshi; Okuno, Yusuke; Harada, Masahide; Takanari, Hiroki; Sakuma, Ichiro; Kamiya, Kaichiro; Kodama, Itsuo

2010-07-01

Intravenous application of amiodarone is commonly used in the treatment of life-threatening arrhythmias, but the underlying mechanism is not fully understood. The purpose of the present study is to investigate the acute effects of amiodarone on spiral wave (SW) re-entry, the primary organization machinery of ventricular tachycardia/fibrillation (VT/VF), in comparison with lidocaine. A two-dimensional ventricular myocardial layer was obtained from 24 Langendorff-perfused rabbit hearts, and epicardial excitations were analyzed by high-resolution optical mapping. During basic stimulation, amiodarone (5 microM) caused prolongation of action potential duration (APD) by 5.6%-9.1%, whereas lidocaine (15 microM) caused APD shortening by 5.0%-6.4%. Amiodarone and lidocaine reduced conduction velocity similarly. Ventricular tachycardias induced by DC stimulation in the presence of amiodarone were of shorter duration (sustained-VTs >30 s/total VTs: 2/58, amiodarone vs 13/52, control), whereas those with lidocaine were of longer duration (22/73, lidocaine vs 14/58, control). Amiodarone caused prolongation of VT cycle length and destabilization of SW re-entry, which is characterized by marked prolongation of functional block lines, frequent wavefront-tail interactions near the rotation center, and considerable drift, leading to its early annihilation via collision with anatomical boundaries. Spiral wave re-entry in the presence of lidocaine was more stabilized than in control. In the anisotropic ventricular myocardium, amiodarone destabilizes SW re-entry facilitating its early termination. Lidocaine, in contrast, stabilizes SW re-entry resulting in its persistence.

Altered cell-matrix associated ADAM proteins in Alzheimer disease.

PubMed

Gerst, J L; Raina, A K; Pirim, I; McShea, A; Harris, P L; Siedlak, S L; Takeda, A; Petersen, R B; Smith, M A

2000-03-01

Alterations in cell-matrix 'contact' are often related to a disruption of cell cycle regulation and, as such, occur variously in neoplasia. Given the recent findings showing cell cycle alterations in Alzheimer disease, we undertook a study of ADAM-1 and 2 (A Disintegrin And Metalloprotease), developmentally-regulated, integrin-binding, membrane-bound metalloproteases. Our results show that whereas ADAM-1 and 2 are found in susceptible hippocampal neurons in Alzheimer disease, these proteins were not generally increased in similar neuronal populations in younger or age-matched controls except in association with age-related neurofibrillary alterations. This increase in both ADAM-1 and 2 in cases of Alzheimer disease was verified by immunoblot analysis (P < 0.05). An ADAM-induced loss of matrix integration would effectively "reset" the mitotic clock and thereby stimulate re-entry into the cell cycle in neurons in Alzheimer disease. Furthermore, given the importance of integrins in maintaining short-term memory, alterations in ADAM proteins or their proteolytic activity could also play a proximal role in the clinico-pathological manifestations of Alzheimer disease. Copyright 2000 Wiley-Liss, Inc.

Mesh-Based Entry Vehicle and Explosive Debris Re-Contact Probability Modeling

NASA Technical Reports Server (NTRS)

McPherson, Mark A.; Mendeck, Gavin F.

2011-01-01

The risk to a crewed vehicle arising from potential re-contact with fragments from an explosive breakup of any jettisoned spacecraft segments during entry has long sought to be quantified. However, great difficulty lies in efficiently capturing the potential locations of each fragment and their collective threat to the vehicle. The method presented in this paper addresses this problem by using a stochastic approach that discretizes simulated debris pieces into volumetric cells, and then assesses strike probabilities accordingly. Combining spatial debris density and relative velocity between the debris and the entry vehicle, the strike probability can be calculated from the integral of the debris flux inside each cell over time. Using this technique it is possible to assess the risk to an entry vehicle along an entire trajectory as it separates from the jettisoned segment. By decoupling the fragment trajectories from that of the entry vehicle, multiple potential separation maneuvers can then be evaluated rapidly to provide an assessment of the best strategy to mitigate the re-contact risk.

A nontranscriptional role for Oct4 in the regulation of mitotic entry

PubMed Central

Zhao, Rui; Deibler, Richard W.; Lerou, Paul H.; Ballabeni, Andrea; Heffner, Garrett C.; Cahan, Patrick; Unternaehrer, Juli J.; Kirschner, Marc W.; Daley, George Q.

2014-01-01

Rapid progression through the cell cycle and a very short G1 phase are defining characteristics of embryonic stem cells. This distinct cell cycle is driven by a positive feedback loop involving Rb inactivation and reduced oscillations of cyclins and cyclin-dependent kinase (Cdk) activity. In this setting, we inquired how ES cells avoid the potentially deleterious consequences of premature mitotic entry. We found that the pluripotency transcription factor Oct4 (octamer-binding transcription factor 4) plays an unappreciated role in the ES cell cycle by forming a complex with cyclin–Cdk1 and inhibiting Cdk1 activation. Ectopic expression of Oct4 or a mutant lacking transcriptional activity recapitulated delayed mitotic entry in HeLa cells. Reduction of Oct4 levels in ES cells accelerated G2 progression, which led to increased chromosomal missegregation and apoptosis. Our data demonstrate an unexpected nontranscriptional function of Oct4 in the regulation of mitotic entry. PMID:25324523

Injury-activated glial cells promote wound healing of the adult skin in mice.

PubMed

Parfejevs, Vadims; Debbache, Julien; Shakhova, Olga; Schaefer, Simon M; Glausch, Mareen; Wegner, Michael; Suter, Ueli; Riekstina, Una; Werner, Sabine; Sommer, Lukas

2018-01-16

Cutaneous wound healing is a complex process that aims to re-establish the original structure of the skin and its functions. Among other disorders, peripheral neuropathies are known to severely impair wound healing capabilities of the skin, revealing the importance of skin innervation for proper repair. Here, we report that peripheral glia are crucially involved in this process. Using a mouse model of wound healing, combined with in vivo fate mapping, we show that injury activates peripheral glia by promoting de-differentiation, cell-cycle re-entry and dissemination of the cells into the wound bed. Moreover, injury-activated glia upregulate the expression of many secreted factors previously associated with wound healing and promote myofibroblast differentiation by paracrine modulation of TGF-β signalling. Accordingly, depletion of these cells impairs epithelial proliferation and wound closure through contraction, while their expansion promotes myofibroblast formation. Thus, injury-activated glia and/or their secretome might have therapeutic potential in human wound healing disorders.

p53 and Mdm2 act synergistically to maintain cardiac homeostasis and mediate cardiomyocyte cell cycle arrest through a network of microRNAs.

PubMed

Stanley-Hasnain, Shanna; Hauck, Ludger; Grothe, Daniela; Aschar-Sobbi, Roozbeh; Beca, Sanja; Butany, Jagdish; Backx, Peter H; Mak, Tak W; Billia, Filio

2017-01-01

Defining the roadblocks responsible for cell cycle arrest in adult cardiomyocytes lies at the core of developing cardiac regenerative therapies. p53 and Mdm2 are crucial mediators of cell cycle arrest in proliferative cell types, however, little is known about their function in regulating homeostasis and proliferation in terminally differentiated cell types, like cardiomyocytes. To explore this, we generated a cardiac-specific conditional deletion of p53 and Mdm2 (DKO) in adult mice. Herein we describe the development of a dilated cardiomyopathy, in the absence of cardiac hypertrophy. In addition, DKO hearts exhibited a significant increase in cardiomyocyte proliferation. Further evaluation showed that proliferation was mediated by a significant increase in Cdk2 and cyclin E with downregulation of p21 Cip1 and p27 Kip1 . Comparison of miRNA expression profiles from DKO mouse hearts and controls revealed 11 miRNAs that were downregulated in the DKO hearts and enriched for mRNA targets involved in cell cycle regulation. Knockdown of these miRNAs in neonatal rat cardiomyocytes significantly increased cytokinesis with an upregulation in the expression of crucial cell cycle regulators. These results illustrate the importance of the cooperative activities of p53 and Mdm2 in a network of miRNAs that function to impose a barrier against aberrant cardiomyocyte cell cycle re-entry to maintain cardiac homeostasis.

Concise Review: Regeneration in Mammalian Cochlea Hair Cells: Help from Supporting Cells Transdifferentiation.

PubMed

Franco, Bénédicte; Malgrange, Brigitte

2017-03-01

It is commonly assumed that mammalian cochlear cells do not regenerate. Therefore, if hair cells are lost following an injury, no recovery could occur. However, during the first postnatal week, mice harbor some progenitor cells that retain the ability to give rise to new hair cells. These progenitor cells are in fact supporting cells. Upon hair cells loss, those cells are able to generate new hair cells both by direct transdifferentiation or following cell cycle re-entry and differentiation. However, this property of supporting cells is progressively lost after birth. Here, we review the molecular mechanisms that are involved in mammalian hair cell development and regeneration. Manipulating pathways used during development constitute good candidates for inducing hair cell regeneration after injury. Despite these promising studies, there is still no evidence for a recovery following hair cells loss in adult mammals. Stem Cells 2017;35:551-556. © 2017 AlphaMed Press.

The MYC Road to Hearing Restoration

PubMed Central

Kopecky, Benjamin; Fritzsch, Bernd

2012-01-01

Current treatments for hearing loss, the most common neurosensory disorder, do not restore perfect hearing. Regeneration of lost organ of Corti hair cells through forced cell cycle re-entry of supporting cells or through manipulation of stem cells, both avenues towards a permanent cure, require a more complete understanding of normal inner ear development, specifically the balance of proliferation and differentiation required to form and to maintain hair cells. Direct successful alterations to the cell cycle result in cell death whereas regulation of upstream genes is insufficient to permanently alter cell cycle dynamics. The Myc gene family is uniquely situated to synergize upstream pathways into downstream cell cycle control. There are three Mycs that are embedded within the Myc/Max/Mad network to regulate proliferation. The function of the two ear expressed Mycs, N-Myc and L-Myc were unknown less than two years ago and their therapeutic potentials remain speculative. In this review, we discuss the roles the Mycs play in the body and what led us to choose them to be our candidate gene for inner ear therapies. We will summarize the recently published work describing the early and late effects of N-Myc and L-Myc on hair cell formation and maintenance. Lastly, we detail the translational significance of our findings and what future work must be performed to make the ultimate hearing aid: the regeneration of the organ of Corti. PMID:24710525

Two inhibitory systems and CKIs regulate cell cycle exit of mammalian cardiomyocytes after birth

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tane, Shoji; Okayama, Hitomi; Ikenishi, Aiko

Mammalian cardiomyocytes actively proliferate during embryonic stages, following which they exit their cell cycle after birth, and the exit is maintained. Previously, we showed that two inhibitory systems (the G1-phase inhibitory system: repression of cyclin D1 expression; the M-phase inhibitory system: inhibition of CDK1 activation) maintain the cell cycle exit of mouse adult cardiomyocytes. We also showed that two CDK inhibitors (CKIs), p21{sup Cip1} and p27{sup Kip1}, regulate the cell cycle exit in a portion of postnatal cardiomyocytes. It remains unknown whether the two inhibitory systems are involved in the cell cycle exit of postnatal cardiomyocytes and whether p21{sup Cip1}more » and p27{sup Kip1} also inhibit entry to M-phase. Here, we showed that more than 40% of cardiomyocytes entered an additional cell cycle by induction of cyclin D1 expression at postnatal stages, but M-phase entry was inhibited in the majority of cardiomyocytes. Marked cell cycle progression and endoreplication were observed in cardiomyocytes of p21{sup Cip1} knockout mice at 4 weeks of age. In addition, tri- and tetranucleated cardiomyocytes increased significantly in p21{sup Cip1} knockout mice. These data showed that the G1-phase inhibitory system and two CKIs (p21{sup Cip1} and p27{sup Kip1}) inhibit entry to an additional cell cycle in postnatal cardiomyocytes, and that the M-phase inhibitory system and p21{sup Cip1} inhibit M-phase entry of cardiomyocytes which have entered the additional cell cycle. - Highlights: • Many postnatal cardiomyocytes entered an additional cell cycle by cyclin D1 induction. • The majority of cardiomyocytes could not enter M-phase after cyclin D1 induction. • Cell cycle progressed markedly in p21{sup Cip1} knockout mice after postnatal day 14. • Tri- and tetranucleated cardiomyocytes increased in p21{sup Cip1} knockout mice.« less

DCAF1 controls T-cell function via p53-dependent and -independent mechanisms.

PubMed

Guo, Zengli; Kong, Qing; Liu, Cui; Zhang, Song; Zou, Liyun; Yan, Feng; Whitmire, Jason K; Xiong, Yue; Chen, Xian; Wan, Yisong Y

2016-01-05

On activation, naive T cells grow in size and enter cell cycle to mount immune response. How the fundamental processes of T-cell growth and cell cycle entry are regulated is poorly understood. Here we report that DCAF1 (Ddb1-cullin4-associated-factor 1) is essential for these processes. The deletion of DCAF1 in T cells impairs their peripheral homeostasis. DCAF1 is upregulated on T-cell receptor activation and critical for activation-induced T-cell growth, cell cycle entry and proliferation. In addition, DCAF1 is required for T-cell expansion and function during anti-viral and autoimmune responses in vivo. DCAF1 deletion leads to a drastic stabilization of p53 protein, which can be attributed to a requirement of DCAF1 for MDM2-mediated p53 poly-ubiquitination. Importantly, p53 deletion rescues the cell cycle entry defect but not the growth defect of DCAF1-deficient cells. Therefore, DCAF1 is vital for T-cell function through p53-dependent and -independent mechanisms.

DCAF1 controls T-cell function via p53-dependent and -independent mechanisms

PubMed Central

Guo, Zengli; Kong, Qing; Liu, Cui; Zhang, Song; Zou, Liyun; Yan, Feng; Whitmire, Jason K.; Xiong, Yue; Chen, Xian; Wan, Yisong Y.

2016-01-01

On activation, naive T cells grow in size and enter cell cycle to mount immune response. How the fundamental processes of T-cell growth and cell cycle entry are regulated is poorly understood. Here we report that DCAF1 (Ddb1–cullin4-associated-factor 1) is essential for these processes. The deletion of DCAF1 in T cells impairs their peripheral homeostasis. DCAF1 is upregulated on T-cell receptor activation and critical for activation-induced T-cell growth, cell cycle entry and proliferation. In addition, DCAF1 is required for T-cell expansion and function during anti-viral and autoimmune responses in vivo. DCAF1 deletion leads to a drastic stabilization of p53 protein, which can be attributed to a requirement of DCAF1 for MDM2-mediated p53 poly-ubiquitination. Importantly, p53 deletion rescues the cell cycle entry defect but not the growth defect of DCAF1-deficient cells. Therefore, DCAF1 is vital for T-cell function through p53-dependent and -independent mechanisms. PMID:26728942

Keratin 17 modulates hair follicle cycling in a TNFα-dependent fashion

PubMed Central

Tong, Xuemei; Coulombe, Pierre A.

2006-01-01

Mammalian hair follicles cycle between stages of rapid growth (anagen) and metabolic quiescence (telogen) throughout life. Transition from anagen to telogen involves an intermediate stage, catagen, consisting of a swift, apoptosis-driven involution of the lower half of the follicle. How catagen is coordinated, and spares the progenitor cells needed for anagen re-entry, is poorly understood. Keratin 17 (K17)-null mice develop alopecia in the first week post-birth, correlating with hair shaft fragility and untimely apoptosis in the hair bulb. Here we show that this abnormal apoptosis reflects premature entry into catagen. Of the proapoptotic challenges tested, K17-null skin keratinocytes in primary culture are selectively more sensitive to TNFα. K17 interacts with TNF receptor 1 (TNFR1)-associated death domain protein (TRADD), a death adaptor essential for TNFR1-dependent signal relay, suggesting a functional link between this keratin and TNFα signaling. The activity of NF-κB, a downstream target of TNFα, is increased in K17-null skin. We also find that TNFα is required for a timely anagen–catagen transition in mouse pelage follicles, and that its ablation partially rescues the hair cycling defect of K17-null mice. These findings identify K17 and TNFα as two novel and interdependent regulators of hair cycling. PMID:16702408

Changes in the Proliferative Program Limit Astrocyte Homeostasis in the Aged Post-Traumatic Murine Cerebral Cortex.

PubMed

Heimann, Gábor; Canhos, Luisa L; Frik, Jesica; Jäger, Gabriele; Lepko, Tjasa; Ninkovic, Jovica; Götz, Magdalena; Sirko, Swetlana

2017-08-01

Aging leads to adverse outcomes after traumatic brain injury. The mechanisms underlying these defects, however, are not yet clear. In this study, we found that astrocytes in the aged post-traumatic cerebral cortex develop a significantly reduced proliferative response, resulting in reduced astrocyte numbers in the penumbra. Moreover, experiments of reactive astrocytes in vitro reveal that their diminished proliferation is due to an age-related switch in the division mode with reduced cell-cycle re-entry rather than changes in cell-cycle length. Notably, reactive astrocytes in vivo and in vitro become refractory to stimuli increasing their proliferation during aging, such as Sonic hedgehog signaling. These data demonstrate for the first time that age-dependent, most likely intrinsic changes in the proliferative program of reactive astrocytes result in their severely hampered proliferative response to traumatic injury thereby affecting astrocyte homeostasis. © The Author 2017. Published by Oxford University Press.

Re-entry simulation chamber for thermo-mechanical characterisation of space materials

NASA Astrophysics Data System (ADS)

Liedtke, Volker

2003-09-01

During re-entry, materials and components are subject to very high thermal and mechanical loads. Any failure may cause loss of mission. Therefore, materials and components have to be tested under most rigid conditions to verify the suitability of the material and to verify the design of the components. The Re-Entry Simulation Chamber (RESiC) at ARC Seibersdorf research (ARCS) allows simulating the high thermal loads as well as complex mechanical load profiles that may occur during a re-entry; additionally, the influence of chemical reactions of materials with gaseous components of the atmosphere can be studied. The high vacuum chamber (better than 1×10-6 mbar) has a diameter of 650 mm and allows a sample height of 500 mm, or 1000 mm with extension flange. The gas dosing system is designed to emulate the increasing atmospheric pressure during the re-entry trajectory of a vehicle. Heating is performed by a 30 kW induction generator that allows a sufficiently rapid heating of larger components; electrically conductive materials such as metals or carbon fibre reinforced ceramics are directly heated, while for electrical insulators, susceptor plates or tubes will be employed. The uniaxial servo-hydraulic testing machine has a maximum load of 70 kN, either static or with a frequency of up to 70 Hz, with any given load profile (sinus, rectangular, triangular, ...). Strain measurements will be done by non-contacting laser speckle system for maximum flexibility and minimum instrumentation time effort (currently under application testing), or by strain gauges. All relevant process parameters are controlled and recorded by microcomputer. The highly sophisticated control software allows a convenient and reliable multi-channel data acquisition, e.g. temperatures at various positions of the test piece, pressure, loads, strains, and any other test data according to customer specifications; the data format is suitable for any further data processing. During the set-up and operation testing, the device has successfully been employed for thermal shock testing, thermal cycling and gas cycling tests, thermomechanical tests and combinations thereof, e.g. sintering or hot-pressing. During the current final test series, the device will be completed, further optimised and shall be fully operational in summer 2003.

p57KIP2 regulates radial glia and intermediate precursor cell cycle dynamics and lower layer neurogenesis in developing cerebral cortex

PubMed Central

Mairet-Coello, Georges; Tury, Anna; Van Buskirk, Elise; Robinson, Kelsey; Genestine, Matthieu; DiCicco-Bloom, Emanuel

2012-01-01

During cerebral cortex development, precise control of precursor cell cycle length and cell cycle exit is required for balanced precursor pool expansion and layer-specific neurogenesis. Here, we defined the roles of cyclin-dependent kinase inhibitor (CKI) p57KIP2, an important regulator of G1 phase, using deletion mutant mice. Mutant mice displayed macroencephaly associated with cortical hyperplasia during late embryogenesis and postnatal development. Embryonically, proliferation of radial glial cells (RGC) and intermediate precursors (IPC) was increased, expanding both populations, with greater effect on IPCs. Furthermore, cell cycle re-entry was increased during early corticogenesis, whereas cell cycle exit was augmented at middle stage. Consequently, neurogenesis was reduced early, whereas it was enhanced during later development. In agreement, the timetable of early neurogenesis, indicated by birthdating analysis, was delayed. Cell cycle dynamics analyses in mutants indicated that p57KIP2 regulates cell cycle length in both RGCs and IPCs. By contrast, related CKI p27KIP1 controlled IPC proliferation exclusively. Furthermore, p57KIP2 deficiency markedly increased RGC and IPC divisions at E14.5, whereas p27KIP1 increased IPC proliferation at E16.5. Consequently, loss of p57KIP2 increased primarily layer 5-6 neuron production, whereas loss of p27KIP1 increased neurons specifically in layers 2-5. In conclusion, our observations suggest that p57KIP2 and p27KIP1 control neuronal output for distinct cortical layers by regulating different stages of precursor proliferation, and support a model in which IPCs contribute to both lower and upper layer neuron generation. PMID:22223678

Atrial fibrillation driven by micro-anatomic intramural re-entry revealed by simultaneous sub-epicardial and sub-endocardial optical mapping in explanted human hearts.

PubMed

Hansen, Brian J; Zhao, Jichao; Csepe, Thomas A; Moore, Brandon T; Li, Ning; Jayne, Laura A; Kalyanasundaram, Anuradha; Lim, Praise; Bratasz, Anna; Powell, Kimerly A; Simonetti, Orlando P; Higgins, Robert S D; Kilic, Ahmet; Mohler, Peter J; Janssen, Paul M L; Weiss, Raul; Hummel, John D; Fedorov, Vadim V

2015-09-14

The complex architecture of the human atria may create physical substrates for sustained re-entry to drive atrial fibrillation (AF). The existence of sustained, anatomically defined AF drivers in humans has been challenged partly due to the lack of simultaneous endocardial-epicardial (Endo-Epi) mapping coupled with high-resolution 3D structural imaging. Coronary-perfused human right atria from explanted diseased hearts (n = 8, 43-72 years old) were optically mapped simultaneously by three high-resolution CMOS cameras (two aligned Endo-Epi views (330 µm2 resolution) and one panoramic view). 3D gadolinium-enhanced magnetic resonance imaging (GE-MRI, 80 µm3 resolution) revealed the atrial wall structure varied in thickness (1.0 ± 0.7-6.8 ± 2.4 mm), transmural fiber angle differences, and interstitial fibrosis causing transmural activation delay from 23 ± 11 to 43 ± 22 ms at increased pacing rates. Sustained AF (>90 min) was induced by burst pacing during pinacidil (30-100 µM) perfusion. Dual-sided sub-Endo-sub-Epi optical mapping revealed that AF was driven by spatially and temporally stable intramural re-entry with 107 ± 50 ms cycle length and transmural activation delay of 67 ± 31 ms. Intramural re-entrant drivers were captured primarily by sub-Endo mapping, while sub-Epi mapping visualized re-entry or 'breakthrough' patterns. Re-entrant drivers were anchored on 3D micro-anatomic tracks (15.4 ± 2.2 × 6.0 ± 2.3 mm2, 2.9 ± 0.9 mm depth) formed by atrial musculature characterized by increased transmural fiber angle differences and interstitial fibrosis. Targeted radiofrequency ablation of the tracks verified these re-entries as drivers of AF. Integrated 3D structural-functional mapping of diseased human right atria ex vivo revealed that the complex atrial microstructure caused significant differences between Endo vs. Epi activation during pacing and sustained AF driven by intramural re-entry anchored to fibrosis-insulated atrial bundles. Published on behalf of the European Society of Cardiology. All rights reserved. © The Author 2015. For permissions please email: journals.permissions@oup.com.

Atrial fibrillation driven by micro-anatomic intramural re-entry revealed by simultaneous sub-epicardial and sub-endocardial optical mapping in explanted human hearts

PubMed Central

Hansen, Brian J.; Zhao, Jichao; Csepe, Thomas A.; Moore, Brandon T.; Li, Ning; Jayne, Laura A.; Kalyanasundaram, Anuradha; Lim, Praise; Bratasz, Anna; Powell, Kimerly A.; Simonetti, Orlando P.; Higgins, Robert S.D.; Kilic, Ahmet; Mohler, Peter J.; Janssen, Paul M.L.; Weiss, Raul; Hummel, John D.; Fedorov, Vadim V.

2015-01-01

Aims The complex architecture of the human atria may create physical substrates for sustained re-entry to drive atrial fibrillation (AF). The existence of sustained, anatomically defined AF drivers in humans has been challenged partly due to the lack of simultaneous endocardial–epicardial (Endo–Epi) mapping coupled with high-resolution 3D structural imaging. Methods and results Coronary-perfused human right atria from explanted diseased hearts (n = 8, 43–72 years old) were optically mapped simultaneously by three high-resolution CMOS cameras (two aligned Endo–Epi views (330 µm2 resolution) and one panoramic view). 3D gadolinium-enhanced magnetic resonance imaging (GE-MRI, 80 µm3 resolution) revealed the atrial wall structure varied in thickness (1.0 ± 0.7–6.8 ± 2.4 mm), transmural fiber angle differences, and interstitial fibrosis causing transmural activation delay from 23 ± 11 to 43 ± 22 ms at increased pacing rates. Sustained AF (>90 min) was induced by burst pacing during pinacidil (30–100 µM) perfusion. Dual-sided sub-Endo–sub-Epi optical mapping revealed that AF was driven by spatially and temporally stable intramural re-entry with 107 ± 50 ms cycle length and transmural activation delay of 67 ± 31 ms. Intramural re-entrant drivers were captured primarily by sub-Endo mapping, while sub-Epi mapping visualized re-entry or ‘breakthrough’ patterns. Re-entrant drivers were anchored on 3D micro-anatomic tracks (15.4 ± 2.2 × 6.0 ± 2.3 mm2, 2.9 ± 0.9 mm depth) formed by atrial musculature characterized by increased transmural fiber angle differences and interstitial fibrosis. Targeted radiofrequency ablation of the tracks verified these re-entries as drivers of AF. Conclusions Integrated 3D structural–functional mapping of diseased human right atria ex vivo revealed that the complex atrial microstructure caused significant differences between Endo vs. Epi activation during pacing and sustained AF driven by intramural re-entry anchored to fibrosis-insulated atrial bundles. PMID:26059724

Investigations of Control Surface Seals for Re-entry Vehicles

NASA Technical Reports Server (NTRS)

Dunlap, Patrick H., Jr.; Steinetz, Bruce M.; Curry, Donald M.; DeMange, Jeffrey J.; Rivers, H. Kevin; Hsu, Su-Yuen

2002-01-01

Re-entry vehicles generally require control surfaces (e.g., rudders, body flaps) to steer them during flight. Control surface seals are installed along hinge lines and where control surface edges move close to the vehicle body. These seals must operate at high temperatures and limit heat transfer to underlying structures to prevent them from overheating and causing possible loss of vehicle structural integrity. This paper presents results for thermal analyses and mechanical testing conducted on the baseline rudder/fin seal design for the X-38 re-entry vehicle. Exposure of the seals in a compressed state at the predicted peak seal temperature of 1900 F resulted in loss of seal resiliency. The vertical Inconel rudder/fin rub surface was re-designed to account for this loss of resiliency. Room temperature compression tests revealed that seal unit loads and contact pressures were below limits set to protect Shuttle thermal tiles on the horizontal sealing surface. The seals survived an ambient temperature 1000 cycle scrub test over sanded Shuttle tiles and were able to disengage and re-engage the tile edges during testing. Arc jet tests confirmed the need for seals in the rudder/fin gap location because a single seal caused a large temperature drop (delta T = 1710 F) in the gap.

Comparative analysis of ear-hole closure identifies epimorphic regeneration as a discrete trait in mammals

PubMed Central

Gawriluk, Thomas R.; Simkin, Jennifer; Thompson, Katherine L.; Biswas, Shishir K.; Clare-Salzler, Zak; Kimani, John M.; Kiama, Stephen G.; Smith, Jeramiah J.; Ezenwa, Vanessa O.; Seifert, Ashley W.

2016-01-01

Why mammals have poor regenerative ability has remained a long-standing question in biology. In regenerating vertebrates, injury can induce a process known as epimorphic regeneration to replace damaged structures. Using a 4-mm ear punch assay across multiple mammalian species, here we show that several Acomys spp. (spiny mice) and Oryctolagus cuniculus completely regenerate tissue, whereas other rodents including MRL/MpJ ‘healer' mice heal similar injuries by scarring. We demonstrate ear-hole closure is independent of ear size, and closure rate can be modelled with a cubic function. Cellular and genetic analyses reveal that injury induces blastema formation in Acomys cahirinus. Despite cell cycle re-entry in Mus musculus and A. cahirinus, efficient cell cycle progression and proliferation only occurs in spiny mice. Together, our data unite blastema-mediated regeneration in spiny mice with regeneration in other vertebrates such as salamanders, newts and zebrafish, where all healthy adults regenerate in response to injury. PMID:27109826

S phase entry causes homocysteine-induced death while ataxia telangiectasia and Rad3 related protein functions anti-apoptotically to protect neurons.

PubMed

Ye, Weizhen; Blain, Stacy W

2010-08-01

A major phenotype seen in neurodegenerative disorders is the selective loss of neurons due to apoptotic death and evidence suggests that inappropriate re-activation of cell cycle proteins in post-mitotic neurons may be responsible. To investigate whether reactivation of the G1 cell cycle proteins and S phase entry was linked with apoptosis, we examined homocysteine-induced neuronal cell death in a rat cortical neuron tissue culture system. Hyperhomocysteinaemia is a physiological risk factor for a variety of neurodegenerative diseases, including Alzheimer's disease. We found that in response to homocysteine treatment, cyclin D1, and cyclin-dependent kinases 4 and 2 translocated to the nucleus, and p27 levels decreased. Both cyclin-dependent kinases 4 and 2 regained catalytic activity, the G1 gatekeeper retinoblastoma protein was phosphorylated and DNA synthesis was detected, suggesting transit into S phase. Double-labelling immunofluorescence showed a 95% co-localization of anti-bromodeoxyuridine labelling with apoptotic markers, demonstrating that those cells that entered S phase eventually died. Neurons could be protected from homocysteine-induced death by methods that inhibited G1 phase progression, including down-regulation of cyclin D1 expression, inhibition of cyclin-dependent kinases 4 or 2 activity by small molecule inhibitors, or use of the c-Abl kinase inhibitor, Gleevec, which blocked cyclin D and cyclin-dependent kinase 4 nuclear translocation. However, blocking cell cycle progression post G1, using DNA replication inhibitors, did not prevent apoptosis, suggesting that death was not preventable post the G1-S phase checkpoint. While homocysteine treatment caused DNA damage and activated the DNA damage response, its mechanism of action was distinct from that of more traditional DNA damaging agents, such as camptothecin, as it was p53-independent. Likewise, inhibition of the DNA damage sensors, ataxia-telangiectasia mutant and ataxia telangiectasia and Rad3 related proteins, did not rescue apoptosis and in fact exacerbated death, suggesting that the DNA damage response might normally function neuroprotectively to block S phase-dependent apoptosis induction. As cell cycle events appear to be maintained in vivo in affected neurons for weeks to years before apoptosis is observed, activation of the DNA damage response might be able to hold cell cycle-induced death in check.

Ascorbic Acid Efflux and Re-uptake in Endothelial Cells: Maintenance of Intracellular Ascorbate

PubMed Central

May, James M.; Qu, Zhi-chao

2013-01-01

Entry of vitamin C or ascorbate into most tissues requires its movement across the endothelial cell barrier of vessels. If trans-cellular ascorbate movement occurs, then it should be evident as ascorbate efflux from endothelial cells. Cultured EA.926 endothelial cells that had been loaded to about 3.5 mM intracellular ascorbate lost 70–80% of ascorbate to the medium over several hours at 37 °C via a non-saturable process that was insensitive to anion transport inhibitors and thiol reagents. Oxidation of this extracellular ascorbate by ascorbate oxidase or ferricyanide enhanced apparent ascorbate efflux, suggesting that efflux of the vitamin was countered in part by its re-uptake on ascorbate transporters. Although basal ascorbate efflux was not calcium-dependent, increased entry of calcium into the cells enhanced ascorbate release. These results support the hypothesis that ascorbate efflux reflects trans-endothelial cell ascorbate movement out of the blood vessel. PMID:19148707

Ascorbic acid efflux and re-uptake in endothelial cells: maintenance of intracellular ascorbate.

PubMed

May, James M; Qu, Zhi-chao

2009-05-01

Entry of vitamin C or ascorbate into most tissues requires its movement across the endothelial cell barrier of vessels. If trans-cellular ascorbate movement occurs, then it should be evident as ascorbate efflux from endothelial cells. Cultured EA.926 endothelial cells that had been loaded to about 3.5 mM intracellular ascorbate lost 70-80% of ascorbate to the medium over several hours at 37 degrees C via a non-saturable process that was insensitive to anion transport inhibitors and thiol reagents. Oxidation of this extracellular ascorbate by ascorbate oxidase or ferricyanide enhanced apparent ascorbate efflux, suggesting that efflux of the vitamin was countered in part by its re-uptake on ascorbate transporters. Although basal ascorbate efflux was not calcium-dependent, increased entry of calcium into the cells enhanced ascorbate release. These results support the hypothesis that ascorbate efflux reflects trans-endothelial cell ascorbate movement out of the blood vessel.

Cognate interactions between helper T cells and B cells. IV. Requirements for the expression of effector phase activity by helper T cells.

PubMed

Bartlett, W C; McCann, J; Shepherd, D M; Roy, M; Noelle, R J

1990-12-15

After activation with anti-CD3, activated Th (THCD3), but not resting Th, fixed with paraformaldehyde induce B cell RNA synthesis when co-cultured with resting B cells. This activity is expressed by Th of both Th1 and Th2 subtypes, as well as a third Th clone that is not classified into either subtype. It is proposed that anti-CD3 activation of Th results in the expression of Th membrane proteins that trigger B cell cycle entry. Kinetic studies reveal that 4 to 8 h of activation with anti-CD3 is sufficient for ThCD3 to express B cell-activating function. However, activation of Th with anti-CD3 for extended periods of time results in reduced Th effector activity. Inhibition of Th RNA synthesis during the anti-CD3 activation period ablates the ability of ThCD3 to induce B cell cycle entry. This indicates that de novo synthesis of proteins is required for ThCD3 to express effector function. The ability of fixed ThCD3 to induce entry of B cell into cycle is not due to an increase in expression of CD3, CD4, LFA-1, ICAM-1, class I MHC or Thy-1. Other forms of Th activation (PMA and A23187, Con A) also induced Th effector function. Furthermore, purified plasma membranes from anti-CD3 activated, but not resting Th, induced resting B cells to enter cycle. The addition of IL-4, but not IL-2, IL-5, or IFN-gamma amplified the DNA synthetic response of B cells stimulated with PM from activated Th. Taken together these data indicate that de novo expression of Th surface proteins on activated Th is required for Th to induce B cell cycle entry into G1 and the addition of IL-4 is required for the heightened progression into S phase.

Role of the retinoblastoma protein in cell cycle arrest mediated by a novel cell surface proliferation inhibitor

NASA Technical Reports Server (NTRS)

Enebo, D. J.; Fattaey, H. K.; Moos, P. J.; Johnson, T. C.; Spooner, B. S. (Principal Investigator)

1994-01-01

A novel cell regulatory sialoglycopeptide (CeReS-18), purified from the cell surface of bovine cerebral cortex cells has been shown to be a potent and reversible inhibitor of proliferation of a wide array of fibroblasts as well as epithelial-like cells and nontransformed and transformed cells. To investigate the possible mechanisms by which CeReS-18 exerts its inhibitory action, the effect of the inhibitor on the posttranslational regulation of the retinoblastoma susceptibility gene product (RB), a tumor suppressor gene, has been examined. It is shown that CeReS-18 mediated cell cycle arrest of both human diploid fibroblasts (HSBP) and mouse fibroblasts (Swiss 3T3) results in the maintenance of the RB protein in the hypophosphorylated state, consistent with a late G1 arrest site. Although their normal nontransformed counterparts are sensitive to cell cycle arrest mediated by CeReS-18, cell lines lacking a functional RB protein, through either genetic mutation or DNA tumor virus oncoprotein interaction, are less sensitive. The refractory nature of these cells is shown to be independent of specific surface receptors for the inhibitor, and another tumor suppressor gene (p53) does not appear to be involved in the CeReS-18 inhibition of cell proliferation. The requirement for a functional RB protein product, in order for CeReS-18 to mediate cell cycle arrest, is discussed in light of regulatory events associated with density-dependent growth inhibition.

Robust mitotic entry is ensured by a latching switch.

PubMed

Tuck, Chloe; Zhang, Tongli; Potapova, Tamara; Malumbres, Marcos; Novák, Béla

2013-01-01

Cell cycle events are driven by Cyclin dependent kinases (CDKs) and by their counter-acting phosphatases. Activation of the Cdk1:Cyclin B complex during mitotic entry is controlled by the Wee1/Myt1 inhibitory kinases and by Cdc25 activatory phosphatase, which are themselves regulated by Cdk1:Cyclin B within two positive circuits. Impairing these two feedbacks with chemical inhibitors induces a transient entry into M phase referred to as mitotic collapse. The pathology of mitotic collapse reveals that the positive circuits play a significant role in maintaining the M phase state. To better understand the function of these feedback loops during G2/M transition, we propose a simple model for mitotic entry in mammalian cells including spatial control over Greatwall kinase phosphorylation. After parameter calibration, the model is able to recapture the complex and non-intuitive molecular dynamics reported by Potapova et al. (Potapova et al., 2011). Moreover, it predicts the temporal patterns of other mitotic regulators which have not yet been experimentally tested and suggests a general design principle of cell cycle control: latching switches buffer the cellular stresses which accompany cell cycle processes to ensure that the transitions are smooth and robust.

Procedural and Early Outcomes of Two Re-entry Devices for Subintimal Recanalization of Aortoiliac and Femoropopliteal Chronic Total Occlusions.

PubMed

Vuruskan, Ertan; Saracoglu, Erhan

2017-01-01

Subintimal angioplasty is a common treatment choice for chronic total occlusions (CTO) in the iliac and femoropopliteal arteries. This article describes the technical aspects and early outcomes of two different re-entry devices and comparison with manual re-entry technique. A retrospective review of 61 patients (re-entry group) treated with Outback or Pioneer Plus catheters was carried out. A matched cohort of patients (n=62) who underwent lower extremity interventions without the use of re-entry devices (manual re-entry group) were also analyzed (overall 123 patients were analyzed). Procedural success, procedural durations, patency estimates, ankle-brachial indices, and complications were analyzed. Sixty-one patients underwent Outback or Pioneer Plus guided subintimal recanalization. After the procedure, ankle-brachial indices significantly increased in all patients during follow-up. Primary patency for the entire cohort was 83% in the first month. When the re-entry device group was compared with manual re-entry group, no difference was found with respect to success, complication, and patencies between the two groups during follow-up. However, procedure duration and the amount of contrast agent used was significantly decreased in re-entry groups (p<0.001). Also, re-entry time was significantly decreased in Pioneer plus group according to Outback group (p<0.001). Recanalization of CTO using re-entry devices for aortoiliac or femoropopliteal arteries is safe and effective. These devices shorten the procedure time, the re-entry time, reduce radiation risk, and reduce the amount of contrast agent employed.

"Till Death Do Us Part": A Potential Irreversible Link Between Aberrant Cell Cycle Control and Neurodegeneration in the Adult Olfactory Bulb.

PubMed

Omais, Saad; Jaafar, Carine; Ghanem, Noël

2018-01-01

Adult neurogenesis (AN) is an ongoing developmental process that generates newborn neurons in the olfactory bulb (OB) and the hippocampus (Hi) throughout life and significantly contributes to brain plasticity. Adult neural stem and progenitor cells (aNSPCs) are relatively limited in number and fate and are spatially restricted to the subventricular zone (SVZ) and the subgranular zone (SGZ). During AN, the distinct roles played by cell cycle proteins extend beyond cell cycle control and constitute key regulatory mechanisms involved in neuronal maturation and survival. Importantly, aberrant cell cycle re-entry (CCE) in post-mitotic neurons has been strongly linked to the abnormal pathophysiology in rodent models of neurodegenerative diseases with potential implications on the etiology and progression of such diseases in humans. Here, we present an overview of AN in the SVZ-OB and olfactory epithelium (OE) in mice and humans followed by a comprehensive update of the distinct roles played by cell cycle proteins including major tumors suppressor genes in various steps during neurogenesis. We also discuss accumulating evidence underlining a strong link between abnormal cell cycle control, olfactory dysfunction and neurodegeneration in the adult and aging brain. We emphasize that: (1) CCE in post-mitotic neurons due to loss of cell cycle suppression and/or age-related insults as well as DNA damage can anticipate the development of neurodegenerative lesions and protein aggregates, (2) the age-related decline in SVZ and OE neurogenesis is associated with compensatory pro-survival mechanisms in the aging OB which are interestingly similar to those detected in Alzheimer's disease and Parkinson's disease in humans, and (3) the OB represents a well suitable model to study the early manifestation of age-related defects that may eventually progress into the formation of neurodegenerative lesions and, possibly, spread to the rest of the brain. Such findings may provide a novel approach to the modeling of neurodegenerative diseases in humans from early detection to progression and treatment as well.

Oxidative stress triggers cytokinesis failure in hepatocytes upon isolation.

PubMed

Tormos, A M; Taléns-Visconti, R; Bonora-Centelles, A; Pérez, S; Sastre, J

2015-01-01

Primary hepatocytes are highly differentiated cells and proliferatively quiescent. However, the stress produced during liver digestion seems to activate cell cycle entry by proliferative/dedifferentiation programs that still remain unclear. The aim of this work was to assess whether the oxidative stress associated with hepatocyte isolation affects cell cycle and particularly cytokinesis, the final step of mitosis. Hepatocytes were isolated from C57BL/6 mice by collagenase perfusion in the absence and presence of N-acetyl cysteine (NAC). Polyploidy, cell cycle, and reactive oxygen species (ROS) were studied by flow cytometry (DNA, phospho-histone 3, and CellROX(®) Deep Red) and Western blotting (cyclins B1 and D1, and proliferating cell nuclear antigen). mRNA expression of cyclins A1, B1, B2, D1, and F by reverse transcription (RT)-PCR was also assessed. Glutathione levels were measured by mass spectrometry. Here we show that hepatocyte isolation enhanced cell cycle entry, increased hepatocyte binucleation, and caused marked glutathione oxidation. Addition of 5 mM NAC to the hepatocyte isolation media prevented glutathione depletion, partially blocked ROS production and cell cycle entry of hepatocytes, and avoided the blockade of mitosis progression, abrogating defective cytokinesis and diminishing the formation of binucleated hepatocytes during isolation. Therefore, addition of NAC to the isolation media decreased the generation of polyploid hepatocytes confirming that oxidative stress occurs during hepatocyte isolation and it is responsible, at least in part, for cytokinesis failure and hepatocyte binucleation.

Fat cells reactivate quiescent neuroblasts via TOR and glial insulin relays in Drosophila.

PubMed

Sousa-Nunes, Rita; Yee, Lih Ling; Gould, Alex P

2011-03-24

Many stem, progenitor and cancer cells undergo periods of mitotic quiescence from which they can be reactivated. The signals triggering entry into and exit from this reversible dormant state are not well understood. In the developing Drosophila central nervous system, multipotent self-renewing progenitors called neuroblasts undergo quiescence in a stereotypical spatiotemporal pattern. Entry into quiescence is regulated by Hox proteins and an internal neuroblast timer. Exit from quiescence (reactivation) is subject to a nutritional checkpoint requiring dietary amino acids. Organ co-cultures also implicate an unidentified signal from an adipose/hepatic-like tissue called the fat body. Here we provide in vivo evidence that Slimfast amino-acid sensing and Target of rapamycin (TOR) signalling activate a fat-body-derived signal (FDS) required for neuroblast reactivation. Downstream of this signal, Insulin-like receptor signalling and the Phosphatidylinositol 3-kinase (PI3K)/TOR network are required in neuroblasts for exit from quiescence. We demonstrate that nutritionally regulated glial cells provide the source of Insulin-like peptides (ILPs) relevant for timely neuroblast reactivation but not for overall larval growth. Conversely, ILPs secreted into the haemolymph by median neurosecretory cells systemically control organismal size but do not reactivate neuroblasts. Drosophila thus contains two segregated ILP pools, one regulating proliferation within the central nervous system and the other controlling tissue growth systemically. Our findings support a model in which amino acids trigger the cell cycle re-entry of neural progenitors via a fat-body-glia-neuroblasts relay. This mechanism indicates that dietary nutrients and remote organs, as well as local niches, are key regulators of transitions in stem-cell behaviour.

A theoretical study of the initiation, maintenance and termination of gastric slow wave re-entry.

PubMed

Du, Peng; Paskaranandavadivel, Niranchan; O'Grady, Greg; Tang, Shou-Jiang; Cheng, Leo K

2015-12-01

Gastric slow wave dysrhythmias are associated with motility disorders. Periods of tachygastria associated with slow wave re-entry were recently recognized as one important dysrhythmia mechanism, but factors promoting and sustaining gastric re-entry are currently unknown. This study reports two experimental forms of gastric re-entry and presents a series of multi-scale models that define criteria for slow wave re-entry initiation, maintenance and termination. High-resolution electrical mapping was conducted in porcine and canine models and two spatiotemporal patterns of re-entrant activities were captured: single-loop rotor and double-loop figure-of-eight. Two separate multi-scale mathematical models were developed to reproduce the velocity and entrainment frequency of these experimental recordings. A single-pulse stimulus was used to invoke a rotor re-entry in the porcine model and a figure-of-eight re-entry in the canine model. In both cases, the simulated re-entrant activities were found to be perpetuated by tachygastria that was accompanied by a reduction in the propagation velocity in the re-entrant pathways. The simulated re-entrant activities were terminated by a single-pulse stimulus targeted at the tip of re-entrant wave, after which normal antegrade propagation was restored by the underlying intrinsic frequency gradient. (i) the stability of re-entry is regulated by stimulus timing, intrinsic frequency gradient and conductivity; (ii) tachygastria due to re-entry increases the frequency gradient while showing decreased propagation velocity; (iii) re-entry may be effectively terminated by a targeted stimulus at the core, allowing the intrinsic slow wave conduction system to re-establish itself. © The authors 2014. Published by Oxford University Press on behalf of the Institute of Mathematics and its Applications. All rights reserved.

A theoretical study of the initiation, maintenance and termination of gastric slow wave re-entry

PubMed Central

Du, Peng; Paskaranandavadivel, Niranchan; O’Grady, Greg; Tang, Shou-Jiang; Cheng, Leo K.

2015-01-01

Gastric slow wave dysrhythmias are associated with motility disorders. Periods of tachygastria associated with slow wave re-entry were recently recognized as one important dysrhythmia mechanism, but factors promoting and sustaining gastric re-entry are currently unknown. This study reports two experimental forms of gastric re-entry and presents a series of multi-scale models that define criteria for slow wave re-entry initiation, maintenance and termination. High-resolution electrical mapping was conducted in porcine and canine models and two spatiotemporal patterns of re-entrant activities were captured: single-loop rotor and double-loop figure-of-eight. Two separate multi-scale mathematical models were developed to reproduce the velocity and entrainment frequency of these experimental recordings. A single-pulse stimulus was used to invoke a rotor re-entry in the porcine model and a figure-of-eight re-entry in the canine model. In both cases, the simulated re-entrant activities were found to be perpetuated by tachygastria that was accompanied by a reduction in the propagation velocity in the re-entrant pathways. The simulated re-entrant activities were terminated by a single-pulse stimulus targeted at the tip of re-entrant wave, after which normal antegrade propagation was restored by the underlying intrinsic frequency gradient. Main findings: (i) the stability of re-entry is regulated by stimulus timing, intrinsic frequency gradient and conductivity; (ii) tachygastria due to re-entry increases the frequency gradient while showing decreased propagation velocity; (iii) re-entry may be effectively terminated by a targeted stimulus at the core, allowing the intrinsic slow wave conduction system to re-establish itself. PMID:25552487

Enhancing hair follicle regeneration by nonablative fractional laser: Assessment of irradiation parameters and tissue response.

PubMed

Wu, Yueh-Feng; Wang, Shiou-Han; Wu, Pei-Shan; Fan, Sabrina Mai-Yi; Chiu, Hsien-Yi; Tsai, Tsung-Hua; Lin, Sung-Jan

2015-04-01

Identification of methods to enhance anagen entry can be helpful for alopecia. Recently, nonablative laser has been proposed as a potential treatment for alopecia. However, how the laser parameters affect stem cell activity, hair cycles and the associated side effects have not been well characterized. Here we examine the effects of irradiation parameters of 1,550-nm fractional laser on hair cycles. The dorsal skin of eight-week-old female C57BL/6 mice with hair follicles in synchronized telogen was shaved and irradiated with a 1,550-nm fractional erbium-glass laser (Fraxel RE:STORE (SR1500) Laser System, Solta Medical, U.S.A.) with varied beam energies (5-35 mJ) and beam densities (500-3500 microthermal zones/cm(2) ). The cutaneous changes were evaluated both grossly and histologically. Hair follicle stem cell activity was detected by BrdU incorporation and changes in gene expression were quantified by real-time PCR. Direct thermal injury to hair follicles could be observed early after irradiation, especially at higher beam energy. Anagen induction in the irradiated skin showed an all-or-non change. Anagen induction and ulcer formation were affected by the combination of beam energy and density. The lowest beam energy of 5 mJ failed to promote anagen entry at all beam densities tested. As beam energy increased from 10 mJ to 35 mJ, we found a decreasing trend of beam density that could induce anagen entry within 7-9 days with activation of hair follicle stem cells. Beam density above the pro-regeneration density could lead to ulcers and scarring followed by anagen entry in adjacent skin. Analysis of inflammatory cytokines, including TNF-α, IL-1β, and IL-6, revealed that transient moderate inflammation was associated with anagen induction and intense prolonged inflammation preceded ulcer formation. To avoid side effects of hair follicle injury and scarring, appropriate combination of beam energy and density is required. Parameters outside the therapeutic window can result in either no anagen promotion or ulcer formation. © 2015 Wiley Periodicals, Inc.

Calcium influences sensitivity to growth inhibition induced by a cell surface sialoglycopeptide

NASA Technical Reports Server (NTRS)

Betz, N. A.; Fattaey, H. K.; Johnson, T. C.; Spooner, B. S. (Principal Investigator)

1994-01-01

While studies concerning mitogenic factors have been an important area of research for many years, much less is understood about the mechanisms of action of cell surface growth inhibitors. We have purified an 18 kDa cell surface sialoglycopeptide growth inhibitor (CeReS-18) which can reversibly inhibit the proliferation of diverse cell types. The studies discussed in this article show that three mouse keratinocyte cell lines exhibit sixty-fold greater sensitivity than other fibroblasts and epithelial-like cells to CeReS-18-induced growth inhibition. Growth inhibition induced by CeReS-18 treatment is a reversible process, and the three mouse keratinocyte cell lines exhibited either single or multiple cell cycle arrest points, although a predominantly G0/G1 cell cycle arrest point was exhibited in Swiss 3T3 fibroblasts. The sensitivity of the mouse keratinocyte cell lines to CeReS-18-induced growth inhibition was not affected by the degree of tumorigenic progression in the cell lines and was not due to differences in CeReS-18 binding affinity or number of cell surface receptors per cell. However, the sensitivity of both murine fibroblasts and keratinocytes could be altered by changing the extracellular calcium concentration, such that increased extracellular calcium concentrations resulted in decreased sensitivity to CeReS-18-induced proliferation inhibition. Thus the increased sensitivity of the murine keratinocyte cell lines to CeReS-18 could be ascribed to the low calcium concentration used in their propagation. Studies are currently under way investigating the role of calcium in CeReS-18-induced growth arrest. The CeReS-18 may serve as a very useful tool to study negative growth control and the signal transduction events associated with cell cycling.

Animal Models for Studying the In Vivo Functions of Cell Cycle CDKs.

PubMed

Risal, Sanjiv; Adhikari, Deepak; Liu, Kui

2016-01-01

Multiple Cdks (Cdk4, Cdk6, and Cdk2) and a mitotic Cdk (Cdk1) are involved in cell cycle progression in mammals. Cyclins, Cdk inhibitors, and phosphorylations (both activating and inhibitory) at different cellular levels tightly modulate the activities of these kinases. Based on the results of biochemical studies, it was long believed that different Cdks functioned at specific stages during cell cycle progression. However, deletion of all three interphase Cdks in mice affected cell cycle entry and progression only in certain specialized cells such as hematopoietic cells, beta cells of the pancreas, pituitary lactotrophs, and cardiomyocytes. These genetic experiments challenged the prevailing biochemical model and established that Cdks function in a cell-specific, but not a stage-specific, manner during cell cycle entry and the progression of mitosis. Recent in vivo studies have further established that Cdk1 is the only Cdk that is both essential and sufficient for driving the resumption of meiosis during mouse oocyte maturation. These genetic studies suggest a minimal-essential cell cycle model in which Cdk1 is the central regulator of cell cycle progression. Cdk1 can compensate for the loss of the interphase Cdks by forming active complexes with A-, B-, E-, and D-type Cyclins in a stepwise manner. Thus, Cdk1 plays an essential role in both mitosis and meiosis in mammals, whereas interphase Cdks are dispensable.

Re-Entry of Women to the Labour Market After an Interruption in Employment.

ERIC Educational Resources Information Center

Seear, B. N.

The problems involved in the re-entry of women into employment were studied, and the extent to which there exists a demand for employment for re-entry women was examined. A growing number of women are seeking re-entry in a wide range of income levels. The demand for part-time work appears to exceed supply. Official machinery for assisting re-entry…

Preventing re-entry to foster care.

PubMed

Carnochan, Sarah; Rizik-Baer, Daniel; Austin, Michael J

2013-01-01

Re-entry to foster care generally refers to circumstances in which children who have been discharged from foster care to be reunified with their family of origin, adopted, or provided kinship guardianship are returned to foster care. In the context of the federal performance measurement system, re-entry refers specifically to a return to foster care following an unsuccessful reunification. The federal Children and Family Services Review measures re-entry to foster care with a single indicator, called the permanency of reunification indicator, one of four indicators comprising the reunification composite measure. This review focuses on research related to the re-entry indicator, including the characteristics of children, caregivers and families, as well as case and child welfare services that are associated with a higher or lower risk of re-entry to foster care. Promising post-reunification services designed to prevent re-entry to foster care are described.

Characterization of resistance to rhabdovirus and retrovirus infection in a human myeloid cell line.

PubMed

Boso, Guney; Somia, Nikunj V

2015-01-01

Viruses interact with various permissive and restrictive factors in host cells throughout their replication cycle. Cell lines that are non-permissive to viral infection have been particularly useful in discovering host cell proteins involved in viral life cycles. Here we describe the characterization of a human myeloid leukemia cell line, KG-1, that is resistant to infection by retroviruses and a Rhabdovirus. We show that KG-1 cells are resistant to infection by Vesicular Stomatits Virus as well as VSV Glycoprotein (VSVG) pseudotyped retroviruses due to a defect in binding. Moreover our results indicate that entry by xenotropic retroviral envelope glycoprotein RD114 is impaired in KG-1 cells. Finally we characterize a post- entry block in the early phase of the retroviral life cycle in KG-1 cells that renders the cell line refractory to infection. This cell line will have utility in discovering proteins involved in infection by VSV and HIV-1.

Advanced measurement techniques to characterize thermo-mechanical aspects of solid oxide fuel cells

NASA Astrophysics Data System (ADS)

Malzbender, J.; Steinbrech, R. W.

Advanced characterization methods have been used to analyze the thermo-mechanical behaviour of solid oxide fuel cells in a model stack. The primarily experimental work included contacting studies, sealing of a model stack, thermal and re-oxidation cycling. Also an attempt was made to correlate cell fracture in the stack with pore sizes determined from computer tomography. The contacting studies were carried out using pressure sensitive foils. The load to achieve full contact on anode and cathode side of the cell was assessed and applied in the subsequent model stack test. The stack experiment permitted a detailed analysis of stack compaction during sealing. During steady state operation thermal and re-oxidation cycling the changes in open cell voltage and acoustic emissions were monitored. Significant softening of the sealant material was observed at low temperatures. Heating in the thermal cycling loop of the stack appeared to be less critical than the cooling. Re-oxidation cycling led to significant damage if a critical re-oxidation time was exceeded. Microstructural studies permitted further insight into the re-oxidation mechanism. Finally, the maximum defect size in the cell was determined by computer tomography. A limit of maximum anode stress was estimated and the result correlated this with the failure strength observed during the model stack testing.

Optogenetic manipulation of anatomical re-entry by light-guided generation of a reversible local conduction block.

PubMed

Watanabe, Masaya; Feola, Iolanda; Majumder, Rupamanjari; Jangsangthong, Wanchana; Teplenin, Alexander S; Ypey, Dirk L; Schalij, Martin J; Zeppenfeld, Katja; de Vries, Antoine A F; Pijnappels, Daniël A

2017-03-01

Anatomical re-entry is an important mechanism of ventricular tachycardia, characterized by circular electrical propagation in a fixed pathway. It's current investigative and therapeutic approaches are non-biological, rather unspecific (drugs), traumatizing (electrical shocks), or irreversible (ablation). Optogenetics is a new biological technique that allows reversible modulation of electrical function with unmatched spatiotemporal precision using light-gated ion channels. We therefore investigated optogenetic manipulation of anatomical re-entry in ventricular cardiac tissue. Transverse, 150-μm-thick ventricular slices, obtained from neonatal rat hearts, were genetically modified with lentiviral vectors encoding Ca2+-translocating channelrhodopsin (CatCh), a light-gated depolarizing ion channel, or enhanced yellow fluorescent protein (eYFP) as control. Stable anatomical re-entry was induced in both experimental groups. Activation of CatCh was precisely controlled by 470-nm patterned illumination, while the effects on anatomical re-entry were studied by optical voltage mapping. Regional illumination in the pathway of anatomical re-entry resulted in termination of arrhythmic activity only in CatCh-expressing slices by establishing a local and reversible, depolarization-induced conduction block in the illuminated area. Systematic adjustment of the size of the light-exposed area in the re-entrant pathway revealed that re-entry could be terminated by either wave collision or extinction, depending on the depth (transmurality) of illumination. In silico studies implicated source-sink mismatches at the site of subtransmural conduction block as an important factor in re-entry termination. Anatomical re-entry in ventricular tissue can be manipulated by optogenetic induction of a local and reversible conduction block in the re-entrant pathway, allowing effective re-entry termination. These results provide distinctively new mechanistic insight into re-entry termination and a novel perspective for cardiac arrhythmia management. Published on behalf of the European Society of Cardiology. All rights reserved. © The Author 2017. For permissions, please email: journals.permissions@oup.com.

Murine Denys-Drash syndrome: evidence of podocyte de-differentiation and systemic mediation of glomerulosclerosis.

PubMed

Patek, Charles E; Fleming, Stewart; Miles, Colin G; Bellamy, Christopher O; Ladomery, Michael; Spraggon, Lee; Mullins, John; Hastie, Nicholas D; Hooper, Martin L

2003-09-15

Denys-Drash syndrome (DDS) is caused by dominant mutations of the Wilms' tumour suppressor gene, WT1, and characterized by a nephropathy involving diffuse mesangial sclerosis, male pseudohermaphroditism and/or Wilms' tumourigenesis. Previously, we reported that heterozygosity for the Wt1tmT396 mutation induces DDS in heterozygous and chimeric (Wt1tmT396/+<-->+/+) mice. In the present study, the fate of Wt1 mutant cells in chimeric kidneys was assessed by in situ marker analysis, and immunocytochemistry was used to re-examine the claim that glomerulosclerosis (GS) is caused by loss of WT1 and persistent Pax-2 expression by podocytes. Wt1 mutant cells colonized glomeruli efficiently, including podocytes, but some sclerotic glomeruli contained no detectable Wt1 mutant cells. The development of GS was preceded by widespread loss of ZO-1 signal in podocytes (even in kidneys where <5% of glomeruli contained Wt1 mutant podocytes), increased intra-renal renin expression, and de novo podocyte TGF-beta1 expression, but not podocyte Pax-2 expression or loss of WT1, synaptopodin, alpha-actinin-4 or nephrin expression. However, podocytes in partially sclerotic glomeruli that still expressed WT1 at high levels showed reduced vimentin expression, cell cycle re-entry, and re-expressed desmin, cytokeratin and Pax-2. The results suggest that: (i) GS is not due to loss of WT1 expression by podocytes; (ii) podocyte Pax-2 expression reflects re-expression rather than persistent expression, and is the consequence of GS; (iii) GS is mediated systemically and the mechanism involves activation of the renin-angiotensin system; and (iv) podocytes undergo typical maturational changes but subsequently de-differentiate and revert to an immature phenotype during disease progression.

Induction of quiescence (G0) in bone marrow stromal stem cells enhances their stem cell characteristics.

PubMed

Rumman, Mohammad; Majumder, Abhijit; Harkness, Linda; Venugopal, Balu; Vinay, M B; Pillai, Malini S; Kassem, Moustapha; Dhawan, Jyotsna

2018-05-17

Several studies have suggested that bone marrow stromal steam cells (BMSC) exist in a quiescent state (G0) within the in vivo niche; however, an explicit analysis of the biology of G0 state-BMSC has not been reported. We hypothesized that induction of G0 in BMSC might enhance their stem cell properties. Thus, we induced quiescence in BMSC in vitro by (a) suspension culture in a viscous medium or (b) culture on soft polyacrylamide substrate; and examined their molecular and functional phenotype. Induction of G0 was confirmed by bromo-deoxyuridine (BrdU) labelling and analysis of cell cycle gene expression. Upon reactivation and re-entry into cell cycle, G0 state-BMSC exhibited enhanced clonogenic self-renewal, preferential differentiation into osteoblastic rather than adipocytic cells and increased ectopic bone formation when implanted subcutaneously in vivo in immune-deficient mice, compared to asynchronous proliferating (pre-G0) BMSC. Global gene expression profiling revealed reprogramming of the transcriptome during G0 state including significant alterations in relevant pathways and expression of secreted factors, suggesting altered autocrine and paracrine signaling by G0 state-BMSC and a possible mechanism for enhanced bone formation. G0 state-BMSC might provide a clinically relevant model for understanding the in vivo biology of BMSC. Copyright © 2018. Published by Elsevier B.V.

Comparison of cryoablation with 3D mapping versus conventional mapping for the treatment of atrioventricular re-entrant tachycardia and right-sided paraseptal accessory pathways.

PubMed

Russo, Mario S; Drago, Fabrizio; Silvetti, Massimo S; Righi, Daniela; Di Mambro, Corrado; Placidi, Silvia; Prosperi, Monica; Ciani, Michele; Naso Onofrio, Maria T; Cannatà, Vittorio

2016-06-01

Aim Transcatheter cryoablation is a well-established technique for the treatment of atrioventricular nodal re-entry tachycardia and atrioventricular re-entry tachycardia in children. Fluoroscopy or three-dimensional mapping systems can be used to perform the ablation procedure. The aim of this study was to compare the success rate of cryoablation procedures for the treatment of right septal accessory pathways and atrioventricular nodal re-entry circuits in children using conventional or three-dimensional mapping and to evaluate whether three-dimensional mapping was associated with reduced patient radiation dose compared with traditional mapping. In 2013, 81 children underwent transcatheter cryoablation at our institution, using conventional mapping in 41 children - 32 atrioventricular nodal re-entry tachycardia and nine atrioventricular re-entry tachycardia - and three-dimensional mapping in 40 children - 24 atrioventricular nodal re-entry tachycardia and 16 atrioventricular re-entry tachycardia. Using conventional mapping, the overall success rate was 78.1 and 66.7% in patients with atrioventricular nodal re-entry tachycardia or atrioventricular re-entry tachycardia, respectively. Using three-dimensional mapping, the overall success rate was 91.6 and 75%, respectively (p=ns). The use of three-dimensional mapping was associated with a reduction in cumulative air kerma and cumulative air kerma-area product of 76.4 and 67.3%, respectively (p<0.05). The use of three-dimensional mapping compared with the conventional fluoroscopy-guided method for cryoablation of right septal accessory pathways and atrioventricular nodal re-entry circuits in children was associated with a significant reduction in patient radiation dose without an increase in success rate.

Cross-cultural re-entry for missionaries: a new application for the Dual Process Model.

PubMed

Selby, Susan; Clark, Sheila; Braunack-Mayer, Annette; Jones, Alison; Moulding, Nicole; Beilby, Justin

Nearly half a million foreign aid workers currently work worldwide, including over 140,000 missionaries. During re-entry these workers may experience significant psychological distress. This article positions previous research about psychological distress during re-entry, emphasizing loss and grief. At present there is no identifiable theoretical framework to provide a basis for assessment, management, and prevention of re-entry distress in the clinical setting. The development of theoretical concepts and frameworks surrounding loss and grief including the Dual Process Model (DPM) are discussed. All the parameters of the DPM have been shown to be appropriate for the proposed re-entry model, the Dual Process Model applied to Re-entry (DPMR). It is proposed that the DPMR is an appropriate framework to address the processes and strategies of managing re-entry loss and grief. Possible future clinical applications and limitations of the proposed model are discussed. The DPMR is offered for further validation and use in clinical practice.

A successful retrograde re-entry at aorta using the Outback LTD catheter for a bilateral common iliac artery occlusion.

PubMed

Kim, Tae-Hoon; Ahn, Ji-Hun; Kim, Do-Hoi

2013-05-01

The Outback LTD re-entry catheter system has become a valuable tool for peripheral intervention and it has been widely used for variable peripheral chronic total occlusion (CTO). However, its use in the setting of the aorta was restricted because of concerns of bleeding risks resulting from re-entry puncture or ballooning. This report presents a case of successful re-entry using the Outback LTD Re-Entry Catheter (Cordis, Bridgewater, New Jersy) at the aorta in a patient with bilateral common iliac artery occlusion. Copyright © 2012 Wiley Periodicals, Inc.

Parvovirus-Induced Depletion of Cyclin B1 Prevents Mitotic Entry of Infected Cells

PubMed Central

Adeyemi, Richard O.; Pintel, David J.

2014-01-01

Parvoviruses halt cell cycle progression following initiation of their replication during S-phase and continue to replicate their genomes for extended periods of time in arrested cells. The parvovirus minute virus of mice (MVM) induces a DNA damage response that is required for viral replication and induction of the S/G2 cell cycle block. However, p21 and Chk1, major effectors typically associated with S-phase and G2-phase cell cycle arrest in response to diverse DNA damage stimuli, are either down-regulated, or inactivated, respectively, during MVM infection. This suggested that parvoviruses can modulate cell cycle progression by another mechanism. In this work we show that the MVM-induced, p21- and Chk1-independent, cell cycle block proceeds via a two-step process unlike that seen in response to other DNA-damaging agents or virus infections. MVM infection induced Chk2 activation early in infection which led to a transient S-phase block associated with proteasome-mediated CDC25A degradation. This step was necessary for efficient viral replication; however, Chk2 activation and CDC25A loss were not sufficient to keep infected cells in the sustained G2-arrested state which characterizes this infection. Rather, although the phosphorylation of CDK1 that normally inhibits entry into mitosis was lost, the MVM induced DDR resulted first in a targeted mis-localization and then significant depletion of cyclin B1, thus directly inhibiting cyclin B1-CDK1 complex function and preventing mitotic entry. MVM infection thus uses a novel strategy to ensure a pseudo S-phase, pre-mitotic, nuclear environment for sustained viral replication. PMID:24415942

Parvovirus-induced depletion of cyclin B1 prevents mitotic entry of infected cells.

PubMed

Adeyemi, Richard O; Pintel, David J

2014-01-01

Parvoviruses halt cell cycle progression following initiation of their replication during S-phase and continue to replicate their genomes for extended periods of time in arrested cells. The parvovirus minute virus of mice (MVM) induces a DNA damage response that is required for viral replication and induction of the S/G2 cell cycle block. However, p21 and Chk1, major effectors typically associated with S-phase and G2-phase cell cycle arrest in response to diverse DNA damage stimuli, are either down-regulated, or inactivated, respectively, during MVM infection. This suggested that parvoviruses can modulate cell cycle progression by another mechanism. In this work we show that the MVM-induced, p21- and Chk1-independent, cell cycle block proceeds via a two-step process unlike that seen in response to other DNA-damaging agents or virus infections. MVM infection induced Chk2 activation early in infection which led to a transient S-phase block associated with proteasome-mediated CDC25A degradation. This step was necessary for efficient viral replication; however, Chk2 activation and CDC25A loss were not sufficient to keep infected cells in the sustained G2-arrested state which characterizes this infection. Rather, although the phosphorylation of CDK1 that normally inhibits entry into mitosis was lost, the MVM induced DDR resulted first in a targeted mis-localization and then significant depletion of cyclin B1, thus directly inhibiting cyclin B1-CDK1 complex function and preventing mitotic entry. MVM infection thus uses a novel strategy to ensure a pseudo S-phase, pre-mitotic, nuclear environment for sustained viral replication.

Multiple Re-entry Closures After TEVAR for Ruptured Chronic Post-dissection Thoraco-abdominal Aortic Aneurysm.

PubMed

Kinoshita, R; Ganaha, F; Ito, J; Ohyama, N; Abe, N; Yamazato, T; Munakata, H; Mabuni, K; Kugai, T

2018-01-01

Although thoracic endovascular aortic repair (TEVAR) has become a promising treatment for complicated acute type B dissection, its role in treating chronic post-dissection thoraco-abdominal aortic aneurysm (TAA) is still limited owing to persistent retrograde flow into the false lumen (FL) through abdominal or iliac re-entry tears. A case of chronic post-dissection TAA treatment, in which a dilated descending FL ruptured into the left thorax, is described. The primary entry tear was closed by emergency TEVAR and multiple abdominal re-entries were closed by EVAR. In addition, major re-entries at the detached right renal artery and iliac bifurcation were closed using covered stents. To close re-entries as far as possible, EVAR was carried out using the chimney technique, and additional aortic extenders were placed above the coeliac artery. A few re-entries remained, but complete FL thrombosis of the rupture site was achieved. Follow-up computed tomography showed significant shrinkage of the FL. In treating post-dissection TAA, entry closure by TEVAR is sometimes insufficient, owing to persistent retrograde flow into the FL from abdominal or iliac re-entries. Adjunctive techniques are needed to close these distal re-entries to obtain complete FL exclusion, especially in rupture cases. Recently, encouraging results of complete coverage of the thoraco-abdominal aorta with fenestrated or branched endografts have been reported; however, the widespread employment of such techniques appears to be limited owing to technical difficulties. The present method with multiple re-entry closures using off the shelf and immediately available devices is an alternative for the endovascular treatment of post-dissection TAA, especially in the emergency setting.

"Our commonality is our past:" a qualitative analysis of re-entry community health workers' meaningful experiences.

PubMed

Bedell, Precious; Wilson, John L; White, Ann Marie; Morse, Diane S

Re-entry community health workers (CHWs) are individuals who connect diverse community residents at risk for chronic health issues such as Hepatitis C virus and cardiovascular disease with post-prison healthcare and re-entry services. While the utilization of CHWs has been documented in other marginalized populations, there is little knowledge surrounding the work of re-entry CHWs with individuals released from incarceration. Specifically, CHWs' experiences and perceptions of the uniqueness of their efforts to link individuals to healthcare have not been documented systematically. This study explored what is meaningful to formerly incarcerated CHWs as they work with released individuals. The authors conducted a qualitative thematic analysis of twelve meaningful experiences written by re-entry CHWs employed by the Transitions Clinic Network who attended a CHW training program during a conference in San Francisco, CA. Study participants were encouraged to recount meaningful CHW experiences and motivations for working with re-entry populations in a manner consistent with journal-based qualitative analysis techniques. Narratives were coded using an iterative process and subsequently organized according to themes in ATLAS.ti. Study personnel came to consensus with coding and major themes. The narratives highlighted thought processes and meaning related to re-entry CHWs' work helping patients navigate complex social services for successful re-integration. Six major themes emerged from the analysis: advocacy and support, empathy relating to a personal history of incarceration, giving back, professional satisfaction and responsibilities, resiliency and educational advancement, and experiences of social inequities related to race. Re-entry CHWs described former incarceration, employment, and social justice as sources of meaning for assisting justice-involved individuals receive effective, efficient, and high-quality healthcare. Health clinics for individuals released from incarceration provide a unique setting that links high risk patients to needed care and professionalizes career opportunities for formerly incarcerated re-entry CHWs. The commonality of past correctional involvement is a strong indicator of the meaning and perceived effectiveness re-entry CHWs find in working with individuals leaving prison. Expansion of reimbursable visits with re-entry CHWs in transitions clinics designed for re-entering individuals is worthy of further consideration.

Chk1 and Wee1 kinases coordinate DNA replication, chromosome condensation, and anaphase entry

PubMed Central

Fasulo, Barbara; Koyama, Carol; Yu, Kristina R.; Homola, Ellen M.; Hsieh, Tao S.; Campbell, Shelagh D.; Sullivan, William

2012-01-01

Defects in DNA replication and chromosome condensation are common phenotypes in cancer cells. A link between replication and condensation has been established, but little is known about the role of checkpoints in monitoring chromosome condensation. We investigate this function by live analysis, using the rapid division cycles in the early Drosophila embryo. We find that S-phase and topoisomerase inhibitors delay both the initiation and the rate of chromosome condensation. These cell cycle delays are mediated by the cell cycle kinases chk1 and wee1. Inhibitors that cause severe defects in chromosome condensation and congression on the metaphase plate result in delayed anaphase entry. These delays are mediated by wee1 and are not the result of spindle assembly checkpoint activation. In addition, we provide the first detailed live analysis of the direct effect of widely used anticancer agents (aclarubicin, ICRF-193, VM26, doxorubicin, camptothecin, aphidicolin, hydroxyurea, cisplatin, mechlorethamine and x-rays) on key nuclear and cytoplasmic cell cycle events. PMID:22262459

Trajectory optimization study of a lifting body re-entry vehicle for medium to intermediate range applications

NASA Astrophysics Data System (ADS)

Rizvi, S. Tauqeer ul Islam; Linshu, He; ur Rehman, Tawfiq; Rafique, Amer Farhan

2012-11-01

A numerical optimization study of lifting body re-entry vehicles is presented for nominal as well as shallow entry conditions for Medium and Intermediate Range applications. Due to the stringent requirement of a high degree of accuracy for conventional vehicles, lifting re-entry can be used to attain the impact at the desired terminal flight path angle and speed and thus can potentially improve accuracy of the re-entry vehicle. The re-entry of a medium range and intermediate range vehicles is characterized by very high negative flight path angle and low re-entry speed as compared to a maneuverable re-entry vehicle or a common aero vehicle intended for an intercontinental range. Highly negative flight path angles at the re-entry impose high dynamic pressure as well as heat loads on the vehicle. The trajectory studies are carried out to maximize the cross range of the re-entry vehicle while imposing a maximum dynamic pressure constraint of 350 KPa with a 3 MW/m2 heat rate limit. The maximum normal acceleration and the total heat load experienced by the vehicle at the stagnation point during the maneuver have been computed for the vehicle for possible future conceptual design studies. It has been found that cross range capability of up to 35 km can be achieved with a lifting-body design within the heat rate and the dynamic pressure boundary at normal entry conditions. For shallow entry angle of -20 degree and intermediate ranges a cross range capability of up to 250 km can be attained for a lifting body design with less than 10 percent loss in overall range. The normal acceleration also remains within limits. The lifting-body results have also been compared with wing-body results at shallow entry condition. An hp-adaptive pseudo-spectral method has been used for constrained trajectory optimization.

Foster care re-entry: Exploring the role of foster care characteristics, in-home child welfare services and cross-sector services.

PubMed

Lee, Sangmoo; Jonson-Reid, Melissa; Drake, Brett

2012-09-01

This study seeks to advance our understanding of how modifiable and non-modifiable factors may impact the likelihood of re-entry into foster care. Children who entered foster care for the first time following at least one report of maltreatment and were then reunified were followed from exit to re-entry, age 18 or the end of the study period using longitudinal administrative data. Risk of re-entry was explored according to a range of modifiable and non-modifiable case and service characteristics. Children removed from homes with parents who had multiple risk factors (e.g., no high school diploma, mental health diagnosis, criminal record, or teen parents) or were receiving AFDC prior to entry were more likely to re-enter. The receipt of in-home child welfare services during or after foster care was associated with reduced risk of re-entry. Having the longest placement with a relative was associated with decreased risk of re-entry. In conclusion, both modifiable and non-modifiable factors are associated with re-entry into foster care. Among modifiable factors, services appear to have a particularly strong relationship to re-entry. Our data also suggest that in-home child welfare services provided during and after foster care may be associated with improved long-term permanency after return home. Given the continued import of caregiver risk factors even among reunified families, services provided to support reunification should include attention to caregiver needs outside parenting.

Foster care re-entry: Exploring the role of foster care characteristics, in-home child welfare services and cross-sector services☆

PubMed Central

Lee, Sangmoo; Jonson-Reid, Melissa; Drake, Brett

2013-01-01

This study seeks to advance our understanding of how modifiable and non-modifiable factors may impact the likelihood of re-entry into foster care. Children who entered foster care for the first time following at least one report of maltreatment and were then reunified were followed from exit to re-entry, age 18 or the end of the study period using longitudinal administrative data. Risk of re-entry was explored according to a range of modifiable and non-modifiable case and service characteristics. Children removed from homes with parents who had multiple risk factors (e.g., no high school diploma, mental health diagnosis, criminal record, or teen parents) or were receiving AFDC prior to entry were more likely to re-enter. The receipt of in-home child welfare services during or after foster care was associated with reduced risk of re-entry. Having the longest placement with a relative was associated with decreased risk of re-entry. In conclusion, both modifiable and non-modifiable factors are associated with re-entry into foster care. Among modifiable factors, services appear to have a particularly strong relationship to re-entry. Our data also suggest that in-home child welfare services provided during and after foster care may be associated with improved long-term permanency after return home. Given the continued import of caregiver risk factors even among reunified families, services provided to support reunification should include attention to caregiver needs outside parenting. PMID:23729947

Re-entry using anatomically determined isthmuses: a curable ventricular tachycardia in repaired congenital heart disease.

PubMed

Kapel, Gijsbert F L; Reichlin, Tobias; Wijnmaalen, Adrianus P; Piers, Sebastiaan R D; Holman, Eduard R; Tedrow, Usha B; Schalij, Martin J; Stevenson, William G; Zeppenfeld, Katja

2015-02-01

Ventricular tachycardia (VT) is an important cause of late morbidity and mortality in repaired congenital heart disease. The substrate often includes anatomic isthmuses that can be transected by radiofrequency catheter ablation similar to isthmus block for atrial flutter. This study evaluates the long-term efficacy of isthmus block for treatment of re-entry VT in adults with repaired congenital heart disease. Thirty-four patients (49±13 years; 74% male) with repaired congenital heart disease who underwent radiofrequency catheter ablation of VT in 2 centers were included. Twenty-two (65%) had a preserved left and right ventricular function. Patients were inducible for 1 (interquartile range, 1-2) VT, median cycle length: 295 ms (interquartile range, 242-346). Ablation aimed to transect anatomic isthmuses containing VT re-entry circuit isthmuses. Procedural success was defined as noninducibility of any VT and transection of the anatomic isthmus and was achieved in 25 (74%) patients. During long-term follow-up (46±29 months), all patients with procedural success (18/25 with internal cardiac defibrillators) were free of VT recurrence but 7 of 18 experienced internal cardiac defibrillator-related complications. One patient with procedural success and depressed cardiac function received an internal cardiac defibrillator shock for ventricular fibrillation. None of the 18 patients (12/18 with internal cardiac defibrillators) with complete success and preserved cardiac function experienced any ventricular arrhythmia. In contrast, VT recurred in 4 of 9 patients without procedural success. Four patients died from nonarrhythmic causes. In patients with repaired congenital heart disease with preserved ventricular function and isthmus-dependent re-entry, VT isthmus ablation can be curative. © 2014 American Heart Association, Inc.

“Till Death Do Us Part”: A Potential Irreversible Link Between Aberrant Cell Cycle Control and Neurodegeneration in the Adult Olfactory Bulb

PubMed Central

Omais, Saad; Jaafar, Carine; Ghanem, Noël

2018-01-01

Adult neurogenesis (AN) is an ongoing developmental process that generates newborn neurons in the olfactory bulb (OB) and the hippocampus (Hi) throughout life and significantly contributes to brain plasticity. Adult neural stem and progenitor cells (aNSPCs) are relatively limited in number and fate and are spatially restricted to the subventricular zone (SVZ) and the subgranular zone (SGZ). During AN, the distinct roles played by cell cycle proteins extend beyond cell cycle control and constitute key regulatory mechanisms involved in neuronal maturation and survival. Importantly, aberrant cell cycle re-entry (CCE) in post-mitotic neurons has been strongly linked to the abnormal pathophysiology in rodent models of neurodegenerative diseases with potential implications on the etiology and progression of such diseases in humans. Here, we present an overview of AN in the SVZ-OB and olfactory epithelium (OE) in mice and humans followed by a comprehensive update of the distinct roles played by cell cycle proteins including major tumors suppressor genes in various steps during neurogenesis. We also discuss accumulating evidence underlining a strong link between abnormal cell cycle control, olfactory dysfunction and neurodegeneration in the adult and aging brain. We emphasize that: (1) CCE in post-mitotic neurons due to loss of cell cycle suppression and/or age-related insults as well as DNA damage can anticipate the development of neurodegenerative lesions and protein aggregates, (2) the age-related decline in SVZ and OE neurogenesis is associated with compensatory pro-survival mechanisms in the aging OB which are interestingly similar to those detected in Alzheimer's disease and Parkinson's disease in humans, and (3) the OB represents a well suitable model to study the early manifestation of age-related defects that may eventually progress into the formation of neurodegenerative lesions and, possibly, spread to the rest of the brain. Such findings may provide a novel approach to the modeling of neurodegenerative diseases in humans from early detection to progression and treatment as well. PMID:29593485

A theoretical analysis of anatomical and functional intestinal slow wave re-entry.

PubMed

Du, Peng; O'Grady, Gregory; Cheng, Leo K

2017-07-21

Intestinal bioelectrical slow waves are a key regulator of intestinal motility. Peripheral pacemakers, ectopic initiations and sustained periods of re-entrant activities have all been experimentally observed to be important factors in setting the frequency of intestinal slow waves, but the tissue-level mechanisms underpinning these activities are unclear. This theoretical analysis aimed to define the initiation, maintenance, and termination criteria of two classes of intestinal re-entrant activities: anatomical re-entry and functional re-entry. Anatomical re-entry was modeled in a three-dimensional (3D) cylindrical model, and functional rotor was modeled in a 2D rectangle model. A single-pulse stimulus was used to invoke an anatomical re-entry and a prolonged refractory block was used to invoke the rotor. In both cases, the simulated re-entrant activities operated at frequencies above the baseline entrainment frequency. The anatomical re-entry simulation results demonstrated that a temporary functional refractory block would be required to initiate the re-entrant activity in a single direction around the cylindrical model. The rotor could be terminated by a single-pulse stimulus delivered around the core of the rotor. In conclusion, the simulation results provide the following new insights into the mechanisms of intestinal re-entry: (i) anatomical re-entry is only maintained within a specific range of velocities, outside of which the re-entrant activities become either an ectopic activity or simultaneous activations of the intestinal wall; (ii) a maintained rotor entrained slow waves faster in the antegrade direction than in the retrograde direction. Simulations are shown to be a valuable tool for achieving novel insights into the mechanisms of intestinal slow wave dysrhythmia. Copyright © 2017 Elsevier Ltd. All rights reserved.

Teaching Composition to Re-Entry Students

ERIC Educational Resources Information Center

Foulkes, Natalie; Taines, Beatrice

1978-01-01

Describes the Women's Re-Entry Program at Diablo Valley College which uses structured teaching methods to alleviate the two principal weaknesses found in English compositions written by re-entry women, vagueness and lack of organization. (TP)

Use of a Re-entry Device in Left Subclavian Occlusion: Case Series.

PubMed

Thomas, Wiliam Rhodri; Chick, Christopher; Goyal, Nimit

2018-01-01

To describe the use of a re-entry catheter in the endovascular treatment of left subclavian stenosis. We present three patients where initial attempts at re-vascularisation using standard techniques were unsuccessful. An OUTBACK catheter was employed to facilitate re-entry in these patients. True lumen re-entry was achieved in all patients, leading to successful treatment of all stenoses. There was a lack of filling of the left vertebral artery post-angioplasty in one patient; this was not clinically significant. The case series presented is encouraging for the use of a re-entry catheter in the treatment of subclavian occlusion. In our limited experience this has proved to be a safe technique for use in patients who fail re-vascularisation by standard methods; a larger study is required to confirm this.

Cardiac re-entry dynamics and self-termination in DT-MRI based model of Human Foetal Heart

NASA Astrophysics Data System (ADS)

Biktasheva, Irina V.; Anderson, Richard A.; Holden, Arun V.; Pervolaraki, Eleftheria; Wen, Fen Cai

2018-02-01

The effect of human foetal heart geometry and anisotropy on anatomy induced drift and self-termination of cardiac re-entry is studied here in MRI based 2D slice and 3D whole heart computer simulations. Isotropic and anisotropic models of 20 weeks of gestational age human foetal heart obtained from 100μm voxel diffusion tensor MRI data sets were used in the computer simulations. The fiber orientation angles of the heart were obtained from the orientation of the DT-MRI primary eigenvectors. In a spatially homogeneous electrophysiological monodomain model with the DT-MRI based heart geometries, cardiac re-entry was initiated at a prescribed location in a 2D slice, and in the 3D whole heart anatomy models. Excitation was described by simplified FitzHugh-Nagumo kinetics. In a slice of the heart, with propagation velocity twice as fast along the fibres than across the fibers, DT-MRI based fiber anisotropy changes the re-entry dynamics from pinned to an anatomical re-entry. In the 3D whole heart models, the fiber anisotropy changes cardiac re-entry dynamics from a persistent re-entry to the re-entry self-termination. The self-termination time depends on the re-entry’s initial position. In all the simulations with the DT-MRI based cardiac geometry, the anisotropy of the myocardial tissue shortens the time to re-entry self-termination several folds. The numerical simulations depend on the validity of the DT-MRI data set used. The ventricular wall showed the characteristic transmural rotation of the helix angle of the developed mammalian heart, while the fiber orientation in the atria was irregular.

Simian hemorrhagic fever virus cell entry is dependent on CD163 and uses a clathrin-mediated endocytosis-like pathway.

PubMed

Caì, Yíngyún; Postnikova, Elena N; Bernbaum, John G; Yú, Shu Qìng; Mazur, Steven; Deiuliis, Nicole M; Radoshitzky, Sheli R; Lackemeyer, Matthew G; McCluskey, Adam; Robinson, Phillip J; Haucke, Volker; Wahl-Jensen, Victoria; Bailey, Adam L; Lauck, Michael; Friedrich, Thomas C; O'Connor, David H; Goldberg, Tony L; Jahrling, Peter B; Kuhn, Jens H

2015-01-01

Simian hemorrhagic fever virus (SHFV) causes a severe and almost uniformly fatal viral hemorrhagic fever in Asian macaques but is thought to be nonpathogenic for humans. To date, the SHFV life cycle is almost completely uncharacterized on the molecular level. Here, we describe the first steps of the SHFV life cycle. Our experiments indicate that SHFV enters target cells by low-pH-dependent endocytosis. Dynamin inhibitors, chlorpromazine, methyl-β-cyclodextrin, chloroquine, and concanamycin A dramatically reduced SHFV entry efficiency, whereas the macropinocytosis inhibitors EIPA, blebbistatin, and wortmannin and the caveolin-mediated endocytosis inhibitors nystatin and filipin III had no effect. Furthermore, overexpression and knockout study and electron microscopy results indicate that SHFV entry occurs by a dynamin-dependent clathrin-mediated endocytosis-like pathway. Experiments utilizing latrunculin B, cytochalasin B, and cytochalasin D indicate that SHFV does not hijack the actin polymerization pathway. Treatment of target cells with proteases (proteinase K, papain, α-chymotrypsin, and trypsin) abrogated entry, indicating that the SHFV cell surface receptor is a protein. Phospholipases A2 and D had no effect on SHFV entry. Finally, treatment of cells with antibodies targeting CD163, a cell surface molecule identified as an entry factor for the SHFV-related porcine reproductive and respiratory syndrome virus, diminished SHFV replication, identifying CD163 as an important SHFV entry component. Simian hemorrhagic fever virus (SHFV) causes highly lethal disease in Asian macaques resembling human illness caused by Ebola or Lassa virus. However, little is known about SHFV's ecology and molecular biology and the mechanism by which it causes disease. The results of this study shed light on how SHFV enters its target cells. Using electron microscopy and inhibitors for various cellular pathways, we demonstrate that SHFV invades cells by low-pH-dependent, actin-independent endocytosis, likely with the help of a cellular surface protein. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Hippo pathway coactivators Yap and Taz are required to coordinate mammalian liver regeneration

PubMed Central

Lu, Li; Finegold, Milton J; Johnson, Randy L

2018-01-01

The mammalian liver has a remarkable capacity for repair following injury. Removal of up to two-third of liver mass results in a series of events that include extracellular matrix remodeling, coordinated hepatic cell cycle re-entry, restoration of liver mass and tissue remodeling to return the damaged liver to its normal state. Although there has been considerable advancement of our knowledge concerning the regenerative capacity of the mammalian liver, many outstanding questions remaining, such as: how does the regenerating liver stop proliferating when appropriate mass is restored and how do these mechanisms relate to normal regulation of organ size during development? Hippo pathway has been proposed to be central in mediating both events: organ size control during development and following regeneration. In this report, we examined the role of Yap and Taz, key components of the Hippo pathway in liver organ size regulation, both in the context of development and homeostasis. Our studies reveal that contrary to the current paradigms that Yap/Taz are not required for developmental regulation of liver size but are required for proper liver regeneration. In livers depleted of Yap and Taz, liver mass is elevated in neonates and adults. However, Yap/Taz-depleted livers exhibit profound defects in liver regeneration, including an inability to restore liver mass and to properly coordinate cell cycle entry. Taken together, our results highlight requirements for the Hippo pathway during liver regeneration and indicate that there are additional pathways that cooperate with Hippo signaling to control liver size during development and in the adult. PMID:29303509

Evaluation and Influence of Brachiocephalic Branch Re-entry in Patients With Type A Acute Aortic Dissection.

PubMed

Yasuda, Shota; Imoto, Kiyotaka; Uchida, Keiji; Karube, Norihisa; Minami, Tomoyuki; Goda, Motohiko; Suzuki, Shinichi; Masuda, Munetaka

2016-12-22

Stanford type A acute aortic dissection (A-AAD) extends to the brachiocephalic branches in some patients. After ascending aortic replacement, a remaining re-entry tear in the distal brachiocephalic branches may act as an entry and result in a patent false lumen in the aortic arch. However, the effect of brachiocephalic branch re-entry concomitant with A-AAD remains unknown.Methods and Results:Eighty-five patients with A-AAD who underwent ascending aortic replacement in which both preoperative and postoperative multiple-detector computed tomography (MDCT) scans could be evaluated were retrospectively studied. The presence of a patent false lumen in at least one of the brachiocephalic branches on preoperative MDCT was defined as brachiocephalic branch re-entry, and 41 patients (48%) had this. Postoperatively, 47 of 85 (55%) patients had a patent false lumen in the aortic arch. False lumen remained patent after operation in 34 out of the 41 (83%) patients with brachiocephalic branch re-entry, as compared to that in 13 of the 44 (30%) patients without such re-entry (P<0.001). Brachiocephalic branch re-entry was a significant risk factor for a late increase in the aortic arch diameter greater than 10 mm (P=0.047). Brachiocephalic branch re-entry in patients with A-AAD is related to a patent false lumen in the aortic arch early after ascending aortic replacement and is a risk factor for late aortic arch enlargement.

Predictors of re-entry into the child protection system in Singapore: a cumulative ecological-transactional risk model.

PubMed

Li, Dongdong; Chu, Chi Meng; Ng, Wei Chern; Leong, Wai

2014-11-01

This study examines the risk factors of re-entry for 1,750 child protection cases in Singapore using a cumulative ecological-transactional risk model. Using administrative data, the present study found that the overall percentage of Child Protection Service (CPS) re-entry in Singapore is 10.5% based on 1,750 cases, with a range from 3.9% (within 1 year) to 16.5% (within 8 years after case closure). One quarter of the re-entry cases were observed to occur within 9 months from case closure. Seventeen risk factors, as identified from the extant literature, were tested for their utility to predict CPS re-entry in this study using a series of Cox regression analyses. A final list of seven risk factors (i.e., children's age at entry, case type, case closure result, duration of case, household income, family size, and mother's employment status) was used to create a cumulative risk score. The results supported the cumulative risk model in that higher risk score is related to higher risk of CPS re-entry. Understanding the prevalence of CPS re-entry and the risk factors associated with re-entry is the key to informing practice and policy in a culturally relevant way. The results from this study could then be used to facilitate critical case management decisions in order to enhance positive outcomes of families and children in Singapore's care system. Copyright © 2014 Elsevier Ltd. All rights reserved.

Satellite Re-entry Modeling and Uncertainty Quantification

NASA Astrophysics Data System (ADS)

Horsley, M.

2012-09-01

LEO trajectory modeling is a fundamental aerospace capability and has applications in many areas of aerospace, such as maneuver planning, sensor scheduling, re-entry prediction, collision avoidance, risk analysis, and formation flying. Somewhat surprisingly, modeling the trajectory of an object in low Earth orbit is still a challenging task. This is primarily due to the large uncertainty in the upper atmospheric density, about 15-20% (1-sigma) for most thermosphere models. Other contributions come from our inability to precisely model future solar and geomagnetic activities, the potentially unknown shape, material construction and attitude history of the satellite, and intermittent, noisy tracking data. Current methods to predict a satellite's re-entry trajectory typically involve making a single prediction, with the uncertainty dealt with in an ad-hoc manner, usually based on past experience. However, due to the extreme speed of a LEO satellite, even small uncertainties in the re-entry time translate into a very large uncertainty in the location of the re-entry event. Currently, most methods simply update the re-entry estimate on a regular basis. This results in a wide range of estimates that are literally spread over the entire globe. With no understanding of the underlying distribution of potential impact points, the sequence of impact points predicted by the current methodology are largely useless until just a few hours before re-entry. This paper will discuss the development of a set of the High Performance Computing (HPC)-based capabilities to support near real-time quantification of the uncertainty inherent in uncontrolled satellite re-entries. An appropriate management of the uncertainties is essential for a rigorous treatment of the re-entry/LEO trajectory problem. The development of HPC-based tools for re-entry analysis is important as it will allow a rigorous and robust approach to risk assessment by decision makers in an operational setting. Uncertainty quantification results from the recent uncontrolled re-entry of the Phobos-Grunt satellite will be presented and discussed. This work performed under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344.

Targeted True Lumen Re-Entry With the Outback Catheter: Accuracy, Success, and Complications in 100 Peripheral Chronic Total Occlusions and Systematic Review of the Literature.

PubMed

Kitrou, Panagiotis; Parthipun, Aneeta; Diamantopoulos, Athanasios; Paraskevopoulos, Ioannis; Karunanithy, Narayan; Katsanos, Konstantinos

2015-08-01

To report a single-center experience with the Outback re-entry device for targeted distal true lumen re-entry during subintimal recanalization of chronic total occlusions (CTOs) and compare the results with a systematic review of the literature. Between February 2011 and July 2013, 104 Outback devices were employed in 91 patients (mean age 64±9 years; 57 men) for subintimal recanalization of 100 vessels with CTOs after initial failure of spontaneous reentry. Fifty-two cases involved a retrograde approach to aortoiliac occlusions and 48 were re-entry attempts in infrainguinal CTOs. Outcome measures included complications and technical success, defined as successful targeted re-entry at the preplanned site of the distal true lumen. To evaluate device accuracy, the re-entry distance (between the point of true vessel reconstitution and the eventual re-entry point) was measured. Outback success was 93% (93/100); only 7 cases failed owing to heavy calcification (5/52 aortoiliac vs 2/48 infrainguinal, p=0.44). Re-entry was highly accurate, with a re-entry distance of ~1 cm in both subgroups (1.2±0.1 cm in aortoiliac vs 1.3±0.1 cm in infrainguinal, p=0.40). There were no major and 17 minor complications (9/52 aortoiliac vs 8/48 infrainguinal, p=0.93). Results are in line with the systematic review that identified 11 studies (only 1 randomized trial) involving mostly the femoropopliteal segment (119 aortoiliac and 464 infrainguinal segments). The pooled Outback success rate was 90% (95% confidence interval 85% to 94%) and the pooled complication rate was 4.3% (95% confidence interval 1.6% to 8.3%). The Outback device is safe and has a very high rate of achieving targeted true lumen re-entry, which minimizes the sacrifice of healthy vessel in the aortoiliac and infrainguinal arteries. © The Author(s) 2015.

The R2R3 MYB Transcription Factors FOUR LIPS and MYB88 Regulate Female Reproductive Development

PubMed Central

Lamb, Rebecca S.

2012-01-01

Gamete formation is an important step in the life cycle of sexually reproducing organisms. In flowering plants, haploid spores are formed after the meiotic division of spore mother cells. These spores develop into male and female gametophytes containing gametes after undergoing mitotic divisions. In the female, the megaspore mother cell undergoes meiosis forming four megaspores, of which one is functional and three degenerate. The megaspore then undergoes three mitotic cycles thus generating an embryo sac with eight nuclei. The embryo sac undergoes cellularization to form the mature seven-celled female gametophyte. Entry into and progression through meiosis is essential for megasporogenesis and subsequent megagametogenesis, but control of this process is not well understood. FOUR LIPS (FLP) and its paralogue MYB88, encoding R2R3 MYB transcription factors, have been extensively studied for their role in limiting the terminal division in stomatal development by direct regulation of the expression of cell cycle genes. Here it is demonstrated that FLP and MYB88 also regulate female reproduction. Both FLP and MYB88 are expressed during ovule development and their loss significantly increases the number of ovules produced by the placenta. Despite the presence of excess ovules, single and double mutants exhibit reduced seed set due to reduced female fertility. The sterility results at least in part from defective meiotic entry and progression. Therefore, FLP and MYB88 are important regulators of entry into megasporogenesis, and probably act via the regulation of cell cycle genes. PMID:22915737

Re-Thinking Re-Entry: New Approaches to Supporting Students after Study Abroad

ERIC Educational Resources Information Center

Brubaker, Cate

2017-01-01

While participation in study abroad continues to increase, and both pre-departure and in-country support and interventions have become more robust, the re-entry experience after a program ends still typically takes a back seat to other priorities. Consequently, most students are left to navigate the re-entry transition on their own. This article…

Limitations of the Outback LTD re-entry device in femoropopliteal chronic total occlusions.

PubMed

Shin, Susanna H; Baril, Donald; Chaer, Rabih; Rhee, Robert; Makaroun, Michel; Marone, Luke

2011-05-01

Subintimal recanalization for the treatment of femoropopliteal chronic total occlusions (CTO) occasionally requires re-entry devices to access the true lumen distally, but limited information is available on factors predicting the success or failure of these devices. We evaluated the Outback LTD re-entry device (LuMend, Redwood City, Calif; acquired by Cordis Corp, Miami Lakes, Fla). A retrospective review of patients with femoropopliteal CTO from August 2006 to August 2009 was performed. Age, gender, occlusion length, site of re-entry, and the angle of the aortic bifurcation were recorded. Procedural angiograms were used to assign a calcification score (none, mild, moderate, severe) at the re-entry site. Univariate and multivariate logistic regression analyses were used to identify factors predicting failure of re-entry into the true lumen. Of 249 CTOs treated, the re-entry device was used 52 times (20.9%): 47 superficial femoral artery (SFA) occlusions and 5 combined SFA and popliteal artery occlusions (33 TransAtlantic InterSociety Consensus II type C and 18 type D lesions). Of 48 procedures with available angiograms for review, the target re-entry site was at the adductor canal in 30 (62.5%), the above-knee popliteal artery in 13 (27.1%), behind the knee joint in 4 (8.3%), and the mid-SFA in 2 (4.2%). Patients (54% men) were a mean age of 73.1 years. Re-entry was successful in 34 attempts (64.5%). Causes of failure included inability to re-enter the true lumen in 11 (61.1%), difficulty tracking the device over a wire in 3 (16.7%), acute angle of aortic bifurcation in 2 (11.1%), mechanical failure of the device in 1 (5.6%), and difficulty tracking the device through the lesion in 1 (5.6%). Moderate or severe calcification at the site of re-entry was the only significant predictor of failure (odds ratio, 6.3; 95% confidence interval, 1.45-24.48; P = .01). An aortic bifurcation angle ≥40° did trend toward predicting success (odds ratio, 0.23; 95% confidence interval, 0.05-1.02; P = .054). Although the Outback re-entry device can be successful in extending the applicability of endovascular management to difficult femoropopliteal occlusions, it is not uniformly successful in current clinical practice. Significant calcification at the proposed re-entry site is a strong predictor of failure. Copyright © 2011 Society for Vascular Surgery. Published by Mosby, Inc. All rights reserved.

Pro-arrhythmogenic effects of atrial fibrillation-induced electrical remodelling: insights from the three-dimensional virtual human atria

PubMed Central

Colman, Michael A; Aslanidi, Oleg V; Kharche, Sanjay; Boyett, Mark R; Garratt, Clifford; Hancox, Jules C; Zhang, Henggui

2013-01-01

Chronic atrial fibrillation (AF) is associated with structural and electrical remodelling in the atria, which are associated with a high recurrence of AF. Through biophysically detailed computer modelling, this study investigated mechanisms by which AF-induced electrical remodelling promotes and perpetuates AF. A family of Courtemanche–Ramirez–Nattel variant models of human atrial cell action potentials (APs), taking into account of intrinsic atrial electrophysiological properties, was modified to incorporate various experimental data sets on AF-induced changes of major ionic channel currents (ICaL, IKur, Ito, IK1, IKs, INaCa) and on intracellular Ca2+ handling. The single cell models for control and AF-remodelled conditions were incorporated into multicellular three-dimensional (3D) atrial tissue models. Effects of the AF-induced electrical remodelling were quantified as the changes of AP profile, AP duration (APD) and its dispersion across the atria, and the vulnerability of atrial tissue to the initiation of re-entry. The dynamic behaviour of re-entrant excitation waves in the 3D models was characterised. In our simulations, AF-induced electrical remodelling abbreviated atrial APD non-uniformly across the atria; this resulted in relatively short APDs co-existing with marked regional differences in the APD at junctions of the crista terminalis/pectinate muscle, pulmonary veins/left atrium. As a result, the measured tissue vulnerability to re-entry initiation at these tissue junctions was increased. The AF-induced electrical remodelling also stabilized and accelerated re-entrant excitation waves, leading to rapid and sustained re-entry. Under the AF-remodelled condition, re-entrant scroll waves in the 3D model degenerated into persistent and erratic wavelets, leading to fibrillation. In conclusion, realistic 3D atrial tissue models indicate that AF-induced electrical remodelling produces regionally heterogeneous and shortened APD; these respectively facilitate initiation and maintenance of re-entrant excitation waves. PMID:23732649

Pro-arrhythmogenic effects of atrial fibrillation-induced electrical remodelling: insights from the three-dimensional virtual human atria.

PubMed

Colman, Michael A; Aslanidi, Oleg V; Kharche, Sanjay; Boyett, Mark R; Garratt, Clifford; Hancox, Jules C; Zhang, Henggui

2013-09-01

Chronic atrial fibrillation (AF) is associated with structural and electrical remodelling in the atria, which are associated with a high recurrence of AF. Through biophysically detailed computer modelling, this study investigated mechanisms by which AF-induced electrical remodelling promotes and perpetuates AF. A family of Courtemanche-Ramirez-Nattel variant models of human atrial cell action potentials (APs), taking into account of intrinsic atrial electrophysiological properties, was modified to incorporate various experimental data sets on AF-induced changes of major ionic channel currents (ICaL, IKur, Ito, IK1, IKs, INaCa) and on intracellular Ca(2+) handling. The single cell models for control and AF-remodelled conditions were incorporated into multicellular three-dimensional (3D) atrial tissue models. Effects of the AF-induced electrical remodelling were quantified as the changes of AP profile, AP duration (APD) and its dispersion across the atria, and the vulnerability of atrial tissue to the initiation of re-entry. The dynamic behaviour of re-entrant excitation waves in the 3D models was characterised. In our simulations, AF-induced electrical remodelling abbreviated atrial APD non-uniformly across the atria; this resulted in relatively short APDs co-existing with marked regional differences in the APD at junctions of the crista terminalis/pectinate muscle, pulmonary veins/left atrium. As a result, the measured tissue vulnerability to re-entry initiation at these tissue junctions was increased. The AF-induced electrical remodelling also stabilized and accelerated re-entrant excitation waves, leading to rapid and sustained re-entry. Under the AF-remodelled condition, re-entrant scroll waves in the 3D model degenerated into persistent and erratic wavelets, leading to fibrillation. In conclusion, realistic 3D atrial tissue models indicate that AF-induced electrical remodelling produces regionally heterogeneous and shortened APD; these respectively facilitate initiation and maintenance of re-entrant excitation waves.

Ebola virus host cell entry.

PubMed

Sakurai, Yasuteru

2015-01-01

Ebola virus is an enveloped virus with filamentous structure and causes a severe hemorrhagic fever in human and nonhuman primates. Host cell entry is the first essential step in the viral life cycle, which has been extensively studied as one of the therapeutic targets. A virus factor of cell entry is a surface glycoprotein (GP), which is an only essential viral protein in the step, as well as the unique particle structure. The virus also interacts with a lot of host factors to successfully enter host cells. Ebola virus at first binds to cell surface proteins and internalizes into cells, followed by trafficking through endosomal vesicles to intracellular acidic compartments. There, host proteases process GPs, which can interact with an intracellular receptor. Then, under an appropriate circumstance, viral and endosomal membranes are fused, which is enhanced by major structural changes of GPs, to complete host cell entry. Recently the basic research of Ebola virus infection mechanism has markedly progressed, largely contributed by identification of host factors and detailed structural analyses of GPs. This article highlights the mechanism of Ebola virus host cell entry, including recent findings.

Canal switch and re-entry phenomenon in benign paroxysmal positional vertigo: difference between immediate and delayed occurrence.

PubMed

Dispenza, F; DE Stefano, A; Costantino, C; Rando, D; Giglione, M; Stagno, R; Bennici, E

2015-04-01

This prospective study was designed to evaluate the differences between immediate and delayed canal re-entry of otoliths after therapeutic manoeuvres in patients with benign paroxysmal positional vertigo (BPPV). A total of 196 patients with BPPV were visited and 127 matched our inclusion criteria. The mean age was 54.74 years. The horizontal semicircular canal (HSC) was involved in 30 cases and the posterior semicircular canal (PSC) in 97 patients. Patients with hearing loss in the ear affected by BPPV have a more recurrent form, compared to those with normal hearing. An immediate canal re-entry was recorded in 3 patients with HSC BPPV, all with geotropic nystagmus. In 7 patients with PSC BPPV, the immediate canal re-entry was detected and the delayed form was noted in 5 patients. The patients with the delayed canal re-entry underwent more than 2 previous manoeuvres. The canal re-entry was not related to the manoeuvre performed. The timing of the Dix-Hallpike test to verify the resolution of the BPPV had a significant role in immediate canal re-entry. A recurrence in the follow-up at least one month after treatment was recorded in 20 patients and was more frequent in patients that had canal re-entry. The canal re-entry or canal switch is a clinical entity that should be kept in mind of the neurotologist when approaching BPPV patients. It is important to distinguish it from recurrence when delayed and from manoeuvre failure when immediate. The timing of manoeuvre performing, in particular the final verification test after therapeutic sessions, is important to prevent the immediate reflux of particles into canals.

Factors associated with a second deferral among donors eligible for re-entry after a false-positive screening test for syphilis, HCV, HBV and HIV.

PubMed

Grégoire, Y; Germain, M; Delage, G

2018-05-01

Since 25 May 2010, all donors at our blood centre who tested false-positive for HIV, HBV, HCV or syphilis are eligible for re-entry after further testing. Donors who have a second false-positive screening test, either during qualification for or after re-entry, are deferred for life. This study reports on factors associated with the occurrence of such deferrals. Rates of second false-positive results were compared by year of deferral, transmissible disease marker, gender, age, donor status (new or repeat) and testing platform (same or different) both at qualification for re-entry and afterwards. Chi-square tests were used to compare proportions. Cox regression was used for multivariate analyses. Participation rates in the re-entry programme were 42·1%: 25·6% failed to qualify for re-entry [different platform: 2·7%; same platform: 42·9% (P < 0·0001)]. After re-entry, rates of deferral for second false-positive results were 8·4% after 3 years [different platform: 1·8%; same platform: 21·4% (P < 0·0001)]. Deferral rates were higher for HIV and HCV than for HBV at qualification when tested on the same platform. The risk, when analysed by multivariate analyses, of a second deferral for a false-positive result, both at qualification and 3 years after re-entry, was lower for donors deferred on a different platform; this risk was higher for HIV, HCV and syphilis than for HBV and for new donors if tested on the same platform. Re-entry is more often successful when donors are tested on a testing platform different from the one on which they obtained their first false-positive result. © 2018 International Society of Blood Transfusion.

ATV reentry

NASA Image and Video Library

2012-10-03

ISS033-E-009232 (3 Oct. 2012) --- This still photo taken by the Expedition 33 crew members aboard the International Space Station shows evidence of the fiery plunge through Earth?s atmosphere and the destructive re-entry of the European Automated Transfer Vehicle-3 (ATV-3) spacecraft, also known as ?Edoardo Amaldi.? The end of the ATV took place over a remote swath of the Pacific Ocean where any surviving debris safely splashed down a short time later, at around 1:30 a.m. (GMT) on Oct. 3, thus concluding the highly successful ATV-3 mission. Aboard the craft during re-entry was the Re Entry Breakup Recorder (REBR), a spacecraft ?black box? designed to gather data on vehicle disintegration during re-entry in order to improve future spacecraft re-entry models.

Trajectory Design and Control for the Compton Gamma Ray Observatory Re-Entry

NASA Technical Reports Server (NTRS)

Hoge, Susan; Vaughn, Frank; Bauer, Frank H. (Technical Monitor)

2000-01-01

The Compton Gamma Ray Observatory (CGRO) controlled re-entry operation was successfully conducted in June of 2000. The surviving parts of the spacecraft landed in the Pacific Ocean within the predicted footprint. The design of the maneuvers to control the trajectory to accomplish this re-entry presented several challenges. These challenges included timing and duration of the maneuvers, fuel management, post maneuver position knowledge, collision avoidance with other spacecraft, accounting for the break-up of the spacecraft into several pieces with a wide range of ballistic coefficients, and ensuring that the impact footprint would remain within the desired landing area in the event of contingencies. This paper presents the initial re-entry trajectory design and the evolution of the design into the maneuver sequence used for the re-entry. The paper discusses the constraints on the trajectory design, the modifications made to the initial design and the reasons behind these modifications. Data from the re-entry operation are presented.

The cancer paradigms of mammalian regeneration: can mammals regenerate as amphibians?

PubMed

Sarig, Rachel; Tzahor, Eldad

2017-04-01

Regeneration in mammals is restricted to distinct tissues and occurs mainly by expansion and maturation of resident stem cells. During regeneration, even subtle mutations in the proliferating cells may cause a detrimental effect by eliciting abnormal differentiation or malignant transformation. Indeed, cancer in mammals has been shown to arise through deregulation of stem cells maturation, which often leads to a differentiation block and cell transformation. In contrast, lower organisms such as amphibians retain a remarkable regenerative capacity in various organs, which occurs via de- and re-differentiation of mature cells. Interestingly, regenerating amphibian cells are highly resistant to oncogenic transformation. Therapeutic approaches to improve mammalian regeneration mainly include stem-cell transplantations; but, these have proved unsuccessful in non-regenerating organs such as the heart. A recently developed approach is to induce de-differentiation of mature cardiomyocytes using factors that trigger their re-entry into the cell cycle. This novel approach raises numerous questions regarding the balance between transformation and regeneration induced by de-differentiation of mature mammalian somatic cells. Can this balance be controlled artificially? Do de-differentiated cells acquire the protection mechanisms seen in regenerating cells of lower organisms? Is this model unique to the cardiac tissue, which rarely develops tumors? This review describes regeneration processes in both mammals and lower organisms and, particularly, the ability of regenerating cells to avoid transformation. By comparing the characteristics of mammalian embryonic and somatic cells, we discuss therapeutic strategies of using various cell populations for regeneration. Finally, we describe a novel cardiac regeneration approach and its implications for regenerative medicine. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Factors associated with re-entry to out-of-home care among children in England.

PubMed

Mc Grath-Lone, Louise; Dearden, Lorraine; Harron, Katie; Nasim, Bilal; Gilbert, Ruth

2017-01-01

Exiting and re-entering out-of-home care (OHC) is considered a disruption to permanence which may have long-lasting, negative consequences for children due to a lack of stability and continuity. Each year approximately one-third of children in OHC in England exit, but information is lacking on rates of re-entries and associated factors. Using national administrative data, we calculated rates of re-entry among children exiting OHC from 2007 to 2012, identified key child and care factors associated with re-entry using Cox proportional hazards modelling, and developed a simple probability calculator to estimate which groups of children are most likely to re-enter OHC within three months. Between 2007 and 2012 re-entries to OHC in England decreased (from 23.3% to 14.4% within one year of exit, p<0.001), possibly due to concurrent changes in the way children exited OHC. Overall, more than one-third of children exiting OHC in 2008 re-entered within five years (35.3%, N=4076), but rates of re-entry varied by child and care characteristics including age, ethnicity, mode of exit, and placement stability. Based on these associated factors, we developed a calculator that can estimate the likelihood of rapid re-entry to OHC for a group of children and could be used by social care practitioners or service planners. Our findings provide insight into which groups of children are most likely to re-enter OHC, who may benefit from additional support or ongoing monitoring. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

Vulnerability to re-entry in simulated two-dimensional cardiac tissue: effects of electrical restitution and stimulation sequence.

PubMed

Tran, Diana X; Yang, Ming-Jim; Weiss, James N; Garfinkel, Alan; Qu, Zhilin

2007-12-01

Ventricular fibrillation is a lethal arrhythmia characterized by multiple wavelets usually starting from a single or figure-of-eight re-entrant circuit. Understanding the factors regulating vulnerability to the re-entry is essential for developing effective therapeutic strategies to prevent ventricular fibrillation. In this study, we investigated how pre-existing tissue heterogeneities and electrical restitution properties affect the initiation of re-entry by premature extrastimuli in two-dimensional cardiac tissue models. We studied two pacing protocols for inducing re-entry following the "sinus" rhythm (S1) beat: (1) a single premature (S2) extrastimulus in heterogeneous tissue; (2) two premature extrastimuli (S2 and S3) in homogeneous tissue. In the first case, the vulnerable window of re-entry is determined by the spatial dimension and extent of the heterogeneity, and is also affected by electrical restitution properties and the location of the premature stimulus. The vulnerable window first increases as the action potential duration (APD) difference between the inside and outside of the heterogeneous region increases, but then decreases as this difference increases further. Steeper APD restitution reduces the vulnerable window of re-entry. In the second case, electrical restitution plays an essential role. When APD restitution is flat, no re-entry can be induced. When APD restitution is steep, re-entry can be induced by an S3 over a range of S1S2 intervals, which is also affected by conduction velocity restitution. When APD restitution is even steeper, the vulnerable window is reduced due to collision of the spiral tips.

Simulation of launch and re-entry acceleration profiles for testing of shuttle and unmanned microgravity research payloads

NASA Astrophysics Data System (ADS)

Cassanto, J. M.; Ziserman, H. I.; Chapman, D. K.; Korszun, Z. R.; Todd, P.

Microgravity experiments designed for execution in Get-Away Special canisters, Hitchhiker modules, and Reusable Re-entry Satellites will be subjected to launch and re-entry accelerations. Crew-dependent provisions for preventing acceleration damage to equipment or products will not be available for these payloads during flight; therefore, the effects of launch and re-entry accelerations on all aspects of such payloads must be evaluated prior to flight. A procedure was developed for conveniently simulating the launch and re-entry acceleration profiles of the Space Shuttle (3.3 and 1.7 × g maximum, respectively) and of two versions of NASA's proposed materials research Re-usable Re-entry Satellite (8 × g maximum in one case and 4 × g in the other). By using the 7 m centrifuge of the Gravitational Plant Physiology Laboratory in Philadelphia it was found possible to simulate the time dependence of these 5 different acceleration episodes for payload masses up to 59 kg. A commercial low-cost payload device, the “Materials Dispersion Apparatus” of Instrumentation Technology Associates was tested for (1) integrity of mechanical function, (2) retention of fluid in its compartments, and (3) integrity of products under simulated re-entry g-loads. In particular, the sharp rise from 1 g to maximum g-loading that occurs during re-entry in various unmanned vehicles was successfully simulated, conditions were established for reliable functioning of the MDA, and crystals of 5 proteins suspended in compartments filled with mother liquor were subjected to this acceleration load.

Nuclear receptor TLX prevents retinal dystrophy and recruits the corepressor atrophin1.

PubMed

Zhang, Chun-Li; Zou, Yuhua; Yu, Ruth T; Gage, Fred H; Evans, Ronald M

2006-05-15

During mammalian embryogenesis, precise coordination of progenitor cell proliferation and differentiation is essential for proper organ size and function. The involvement of TLX (NR2E1), an orphan nuclear receptor, has been implicated in ocular development, as Tlx-/- mice exhibit visual impairment. Using genetic and biochemical approaches, we show that TLX modulates retinal progenitor cell proliferation and cell cycle re-entry by directly regulating the expression of Pten and its target cyclin D1. Additionally, TLX finely tunes the progenitor differentiation program by modulating the phospholipase C and mitogen-activated protein kinase (MAPK) pathways and the expression of an array of cell type-specific transcriptional regulators. Consequently, Tlx-/- mice have a dramatic reduction in retina thickness and enhanced generation of S-cones, and develop severe early onset retinal dystrophy. Furthermore, TLX interacts with atrophin1 (Atn1), a corepressor that is involved in human neurodegenerative dentatorubral-pallidoluysian atrophy (DRPLA) and that is essential for development of multiple tissues. Together, these results reveal a molecular strategy by which an orphan nuclear receptor can precisely orchestrate tissue-specific proliferation and differentiation programs to prevent retinal malformation and degeneration.

76 FR 58197 - Pre-Release Community Confinement

Federal Register 2010, 2011, 2012, 2013, 2014

2011-09-20

... for pre-release RRC [Residential Re- entry Centers] placement pursuant to 28 CFR 570.20-22 until such.... For clarity, we also add a parenthetical that explains that the Bureau includes residential re-entry... (including residential re-entry centers); and participation in gainful employment, employment search efforts...

Re-Entry: Managing Cross-Cultural Transitions.

ERIC Educational Resources Information Center

Adler, Nancy J.

1981-01-01

Studied the re-entry process of corporate and governmental employees (N=200) returning to Canada after working overseas. Research found re-entry into the original culture was a more difficult transition than moving to the foreign culture. Home-country managers tended to exhibit xenophobia in assessing the potential and actual effectiveness of…

Preparing International Students for the Re-Entry Transition.

ERIC Educational Resources Information Center

Arthur, Nancy

2003-01-01

Counselors play an integral role in assisting international students to manage cross-cultural transitions. Re-entry counseling can support international students to examine their transition experiences, provide education about re-entry, and help to develop anticipatory coping strategies. An example of a workshop is described as a method of…

Locking Nut with Stress-Distributing Insert

NASA Technical Reports Server (NTRS)

Daniels, Christopher C.

2010-01-01

Reusable holders have been devised for evaluating high-temperature, plasma-resistant re-entry materials, especially fabrics. Typical material samples tested support thermal-protection-system damage repair requiring evaluation prior to re-entry into terrestrial atmosphere. These tests allow evaluation of each material to withstand the most severe predicted re-entry conditions.

A Method for Semi-quantitative Assessment of Exposure to Pesticides of Applicators and Re-entry Workers: An Application in Three Farming Systems in Ethiopia.

PubMed

Negatu, Beyene; Vermeulen, Roel; Mekonnen, Yalemtshay; Kromhout, Hans

2016-07-01

To develop an inexpensive and easily adaptable semi-quantitative exposure assessment method to characterize exposure to pesticide in applicators and re-entry farmers and farm workers in Ethiopia. Two specific semi-quantitative exposure algorithms for pesticides applicators and re-entry workers were developed and applied to 601 farm workers employed in 3 distinctly different farming systems [small-scale irrigated, large-scale greenhouses (LSGH), and large-scale open (LSO)] in Ethiopia. The algorithm for applicators was based on exposure-modifying factors including application methods, farm layout (open or closed), pesticide mixing conditions, cleaning of spraying equipment, intensity of pesticide application per day, utilization of personal protective equipment (PPE), personal hygienic behavior, annual frequency of application, and duration of employment at the farm. The algorithm for re-entry work was based on an expert-based re-entry exposure intensity score, utilization of PPE, personal hygienic behavior, annual frequency of re-entry work, and duration of employment at the farm. The algorithms allowed estimation of daily, annual and cumulative lifetime exposure for applicators, and re-entry workers by farming system, by gender, and by age group. For all metrics, highest exposures occurred in LSGH for both applicators and female re-entry workers. For male re-entry workers, highest cumulative exposure occurred in LSO farms. Female re-entry workers appeared to be higher exposed on a daily or annual basis than male re-entry workers, but their cumulative exposures were similar due to the fact that on average males had longer tenure. Factors related to intensity of exposure (like application method and farm layout) were indicated as the main driving factors for estimated potential exposure. Use of personal protection, hygienic behavior, and duration of employment in surveyed farm workers contributed less to the contrast in exposure estimates. This study indicated that farmers' and farm workers' exposure to pesticides can be inexpensively characterized, ranked, and classified. Our method could be extended to assess exposure to specific active ingredients provided that detailed information on pesticides used is available. The resulting exposure estimates will consequently be used in occupational epidemiology studies in Ethiopia and other similar countries with few resources. © The Author 2016. Published by Oxford University Press on behalf of the British Occupational Hygiene Society.

School re-entry of the pediatric heart transplant recipient.

PubMed

Weil, Constance M; Rodgers, S; Rubovits, S

2006-12-01

Pediatric cardiac transplant has become increasingly frequent in the last decade and survival rates have improved remarkably. Outcome research on this population suggests that the majority of children have the capacity for healthy adaptation although 25-40% have been shown to have some type of psychiatric difficulties. As school plays a major role in these children's lives, early intervention and close liaison with schools is indicated to reduce psychological morbidity, enhance adaptation within the school environment and enhance overall adjustment. This paper proposes a model for a school re-entry program for this population. The school re-entry program is aimed at children who are undergoing cardiac transplant and will be entering or re-entering the school system. They may range in academic age from preschool to college level and have been attending private or public schools with placements in regular education programs, regular education programs with resource support, special education programs, and alternative school programs. Others may not have been attending school because of the severity of their medical condition and have been receiving in-home tutoring. Each child is offered school re-entry assistance by a multi-disciplinary team composed of members from the Cardiology Transplant Service. The re-entry program includes cognitive and psychosocial assessment, liaison with the child's school pre- and post-transplant, academic planning and provision of academic, emotional, and behavioral support before, during, and immediately after transplant, a school re-entry visit, and an ongoing school consultation. The goal is to address issues necessary for a successful school re-entry including appropriate academic placement and support, psychosocial adjustment, education of school personnel and ongoing health needs of the student. The next step is to formally evaluate the efficacy of this program in successful school re-entry.

DNA Damage and Genomic Instability Induced by Inappropriate DNA Re-replication

DTIC Science & Technology

2006-04-01

replication in yeast cells. In the prior reporting period we demonstrated that re-replication induces a rapid and significant decrease in cell viability...repair, DNA replication, checkpoint, cell cycle, yeast , RAD9 16. SECURITY CLASSIFICATION OF: 18. NUMBER OF PAGES 19a. NAME OF RESPONSIBLE PERSON...initiation, our laboratory has been able to conditionally induce varying amounts of re- replication in yeast cells. Effectively, cells enter, but do not

Role Management, Educational Satisfaction, and Role Dynamics in Post-Secondary, Re-entry Women.

ERIC Educational Resources Information Center

Edmondon, Mary Ellen; And Others

1986-01-01

A sample of 42 post-secondary, educational re-entry women completed questionnaires focusing on background status, role dynamics, and satisfaction with their re-entry experience. Results showed no differences between students in a vocational program and those in a traditional, academic program. Role-dynamic variables--but not background-status…

Adaptable Holders for Arc-Jet Screening Candidate Thermal Protection System Repair Materials

NASA Technical Reports Server (NTRS)

Riccio, Joe; Milhoan, Jim D.

2010-01-01

Reusable holders have been devised for evaluating high-temperature, plasma-resistant re-entry materials, especially fabrics. Typical material samples tested support thermal-protection-system damage repair requiring evaluation prior to re-entry into terrestrial atmosphere. These tests allow evaluation of each material to withstand the most severe predicted re-entry conditions.

Report of the Defense Science Board Task Force on Future Strategic Strike Skills

DTIC Science & Technology

2006-03-01

25 Figure 11: Re - entry Systems – Critical Skills...SKILLS___________________________________________________25 Figure 9: Guidance – Critical Skills Figure 10: Re - entry Systems – Critical Skills ICBMS/BASING...ability to survive intercontinental ballistic missile re - entry and then to operate effectively thereafter, is another area where current skills are

Re-Entry, Recruitment, and Retention: A Community Relations Model for Sacramento City College.

ERIC Educational Resources Information Center

White, Maureen E.

Enrollment statistics and projections confirm the importance of focusing community college student recruitment and retention efforts on re-entry students. Re-entry students are a distinct and growing population whose educational requirements often differ from those of younger, traditional students. The literature on adult learners indicates that:…

Paramyxovirus Glycoproteins and the Membrane Fusion Process.

PubMed

Aguilar, Hector C; Henderson, Bryce A; Zamora, J Lizbeth; Johnston, Gunner P

2016-09-01

The family Paramyxoviridae includes many viruses that significantly affect human and animal health. An essential step in the paramyxovirus life cycle is viral entry into host cells, mediated by virus-cell membrane fusion. Upon viral entry, infection results in expression of the paramyxoviral glycoproteins on the infected cell surface. This can lead to cell-cell fusion (syncytia formation), often linked to pathogenesis. Thus membrane fusion is essential for both viral entry and cell-cell fusion and an attractive target for therapeutic development. While there are important differences between viral-cell and cell-cell membrane fusion, many aspects are conserved. The paramyxoviruses generally utilize two envelope glycoproteins to orchestrate membrane fusion. Here, we discuss the roles of these glycoproteins in distinct steps of the membrane fusion process. These findings can offer insights into evolutionary relationships among Paramyxoviridae genera and offer future targets for prophylactic and therapeutic development.

Paramyxovirus Glycoproteins and the Membrane Fusion Process

PubMed Central

Aguilar, Hector C.; Henderson, Bryce A.; Zamora, J. Lizbeth; Johnston, Gunner P.

2016-01-01

The family Paramyxoviridae includes many viruses that significantly affect human and animal health. An essential step in the paramyxovirus life cycle is viral entry into host cells, mediated by virus-cell membrane fusion. Upon viral entry, infection results in expression of the paramyxoviral glycoproteins on the infected cell surface. This can lead to cell-cell fusion (syncytia formation), often linked to pathogenesis. Thus membrane fusion is essential for both viral entry and cell-cell fusion and an attractive target for therapeutic development. While there are important differences between viral-cell and cell-cell membrane fusion, many aspects are conserved. The paramyxoviruses generally utilize two envelope glycoproteins to orchestrate membrane fusion. Here, we discuss the roles of these glycoproteins in distinct steps of the membrane fusion process. These findings can offer insights into evolutionary relationships among Paramyxoviridae genera and offer future targets for prophylactic and therapeutic development. PMID:28138419

Bovine parvovirus uses clathrin-mediated endocytosis for cell entry.

PubMed

Dudleenamjil, Enkhmart; Lin, Chin-Yo; Dredge, Devin; Murray, Byron K; Robison, Richard A; Johnson, F Brent

2010-12-01

Entry events of bovine parvovirus (BPV) were studied. Transmission electron micrographs of infected cells showed virus particles in cytoplasmic vesicles. Chemical inhibitors that block certain aspects of the cellular machinery were employed to assess viral dependency upon those cellular processes. Chlorpromazine, ammonium chloride, chloroquine and bafilamicin A1 were used to inhibit acidification of endosomes and clathrin-associated endocytosis. Nystatin was used as an inhibitor of the caveolae pathway. Cytochalasin D and ML-7 were used to inhibit actin and myosin functions, respectively. Nocodazole and colchicine were employed to inhibit microtubule activity. Virus entry was assessed by measuring viral transcription using real-time PCR, synthesis of capsid protein and assembly of infectious progeny virus in the presence of inhibitor blockage. The results indicated that BPV entry into embryonic bovine trachael cells utilizes endocytosis in clathrin-coated vesicles, is dependent upon acidification, and appears to be associated with actin and microtubule dependency. Evidence for viral entry through caveolae was not obtained. These findings provide a fuller understanding of the early cell-entry events of the replication cycle for members of the genus Bocavirus.

ENTRYSAT: A 3U Cubesat to Study the Re-Entry Atmospheric Environment

NASA Astrophysics Data System (ADS)

Garcia, R. F.; Chaix, J.; Mimoun, D.; EntrySat student Team

2014-04-01

The EntrySat is a 3U CubeSat designed to study the uncontrolled atmospheric re-entry. The project, developed by ISAE in collaboration with ONERA, is funded by CNES and is intended to be launched in January 2016, in the context of the QB50 network. The scientific goal is to relate the kinematics of the satellite with the aerothermodynamic environment during re-entry. In particular, data will be compared with the computations of MUSIC/FAST, a new 6-degree of freedom code developed by ONERA to predict the trajectory of space debris. According to these requirements, the satellite will measure the temperature, pressure, heat flux, and drag force during re-entry, as well as the trajectory and attitude of the satellite. One of the major technological challenges is the retrieval of data during the re-entry phase, which will be based on the Iridium satellite network. The system design is based on the use of commercial COTS components, and is mostly developed by students from ISAE. As such, the EntrySat has an important educational value in the formation of young engineers.

Role of calcium in growth inhibition induced by a novel cell surface sialoglycopeptide

NASA Technical Reports Server (NTRS)

Betz, N. A.; Westhoff, B. A.; Johnson, T. C.; Spooner, B. S. (Principal Investigator)

1995-01-01

Our laboratory has purified an 18 kDa cell surface sialoglycopeptide growth inhibitor (CeReS-18) from intact bovine cerebral cortex cells. Evidence presented here demonstrates that sensitivity to CeReS-18-induced growth inhibition in BALB-c 3T3 cells is influenced by calcium, such that a decrease in the calcium concentration in the growth medium results in an increase in sensitivity to CeReS-18. Calcium did not alter CeReS-18 binding to its cell surface receptor and CeReS-18 does not bind calcium directly. Addition of calcium, but not magnesium, to CeReS-18-inhibited 3T3 cells results in reentry into the cell cycle. A greater than 3-hour exposure to increased calcium is required for escape from CeReS-18-induced growth inhibition. The calcium ionophore ionomycin could partially mimic the effect of increasing extracellular calcium, but thapsigargin was ineffective in inducing escape from growth inhibition. Increasing extracellular calcium 10-fold resulted in an approximately 7-fold increase in total cell-associated 45Ca+2, while free intracellular calcium only increased approximately 30%. However, addition of CeReS-18 did not affect total cell-associated calcium or the increase in total cell-associated calcium observed with an increase in extracellular calcium. Serum addition induced mobilization of intracellular calcium and influx across the plasma membrane in 3T3 cells, and pretreatment of 3T3 cells with CeReS-18 appeared to inhibit these calcium mobilization events. These results suggest that a calcium-sensitive step exists in the recovery from CeReS-18-induced growth inhibition. CeReS-18 may inhibit cell proliferation through a novel mechanism involving altering the intracellular calcium mobilization/regulation necessary for cell cycle progression.

PP2ARts1 is a master regulator of pathways that control cell size

PubMed Central

Zapata, Jessica; Dephoure, Noah; MacDonough, Tracy; Yu, Yaxin; Parnell, Emily J.; Mooring, Meghan; Gygi, Steven P.; Stillman, David J.

2014-01-01

Cell size checkpoints ensure that passage through G1 and mitosis occurs only when sufficient growth has occurred. The mechanisms by which these checkpoints work are largely unknown. PP2A associated with the Rts1 regulatory subunit (PP2ARts1) is required for cell size control in budding yeast, but the relevant targets are unknown. In this paper, we used quantitative proteome-wide mass spectrometry to identify proteins controlled by PP2ARts1. This revealed that PP2ARts1 controls the two key checkpoint pathways thought to regulate the cell cycle in response to cell growth. To investigate the role of PP2ARts1 in these pathways, we focused on the Ace2 transcription factor, which is thought to delay cell cycle entry by repressing transcription of the G1 cyclin CLN3. Diverse experiments suggest that PP2ARts1 promotes cell cycle entry by inhibiting the repressor functions of Ace2. We hypothesize that control of Ace2 by PP2ARts1 plays a role in mechanisms that link G1 cyclin accumulation to cell growth. PMID:24493588

Application of the FADS system on the Re-entry Module

NASA Astrophysics Data System (ADS)

Zhen, Huang

2016-07-01

The aerodynamic model for Flush Air Data Sensing System (FADS) is built based on the surface pressure distribution obtained through the pressure orifices laid on specific positions of the surface,and the flight parameters,such as angle of attack,angle of side-slip,Mach number,free-stream static pressure and dynamic pressure are inferred from the aerodynamic model.The flush air data sensing system (FADS) has been used on several flight tests of aircraft and re-entry vehicle,such as,X-15,space shuttle,F-14,X-33,X-43A and so on. This paper discusses the application of the FADS on the re-entry module with blunt body to obtain high-precision aerodynamic parameters.First of all,a basic theory and operating principle of the FADS is shown.Then,the applications of the FADS on typical aircrafts and re-entry vehicles are described.Thirdly,the application mode on the re-entry module with blunt body is discussed in detail,including aerodynamic simulation,pressure distribution,trajectory reconstruction and the hardware shoule be used,such as flush air data sensing system(FADS),inertial navigation system (INS),data acquisition system,data storage system.Finally,ablunt module re-entry flight test from low earth orbit (LEO) is planned to obtain aerodynamic parameters and amend the aerodynamic model with this FADS system data.The results show that FADS system can be applied widely in re-entry module with blunt bodies.

Mathematical Model of Naive T Cell Division and Survival IL-7 Thresholds.

PubMed

Reynolds, Joseph; Coles, Mark; Lythe, Grant; Molina-París, Carmen

2013-01-01

We develop a mathematical model of the peripheral naive T cell population to study the change in human naive T cell numbers from birth to adulthood, incorporating thymic output and the availability of interleukin-7 (IL-7). The model is formulated as three ordinary differential equations: two describe T cell numbers, in a resting state and progressing through the cell cycle. The third is introduced to describe changes in IL-7 availability. Thymic output is a decreasing function of time, representative of the thymic atrophy observed in aging humans. Each T cell is assumed to possess two interleukin-7 receptor (IL-7R) signaling thresholds: a survival threshold and a second, higher, proliferation threshold. If the IL-7R signaling strength is below its survival threshold, a cell may undergo apoptosis. When the signaling strength is above the survival threshold, but below the proliferation threshold, the cell survives but does not divide. Signaling strength above the proliferation threshold enables entry into cell cycle. Assuming that individual cell thresholds are log-normally distributed, we derive population-average rates for apoptosis and entry into cell cycle. We have analyzed the adiabatic change in homeostasis as thymic output decreases. With a parameter set representative of a healthy individual, the model predicts a unique equilibrium number of T cells. In a parameter range representative of persistent viral or bacterial infection, where naive T cell cycle progression is impaired, a decrease in thymic output may result in the collapse of the naive T cell repertoire.

Exploring Career Decision-Making Experiences of Mexican American Re-Entry Community College Women

ERIC Educational Resources Information Center

Dominguez, Cecilia Sophia

2010-01-01

The purpose of this phenomenological investigation was to increase understanding of the career perspectives of 12 Mexican American, re-entry women who were attending a community college. The questions guiding this investigation were: (a) How do Mexican American re-entry college women describe their career decision-making experiences, (b) What do…

Diversion at re-entry using criminogenic CBT: Review and prototypical program development.

PubMed

Heilbrun, Kirk; Pietruszka, Victoria; Thornewill, Alice; Phillips, Sarah; Schiedel, Rebecca

2017-09-01

Society and the criminal justice system prioritize the reduction of reoffending risk as part of any criminal justice intervention. The Sequential Intercept Model identifies five points of interception at which justice-involved individuals can be diverted into a more rehabilitative alternative: (1) law enforcement/emergency services; (2) booking/initial court hearings; (3) jails/courts; (4) re-entry; and (5) community corrections/community support. The present article focuses on diversion as part of Intercept 5 - re-entry planning and specialized services in the community. We describe the challenges associated with diversion at this stage, and review the relevant research. Next, we describe a "criminogenic cognitive behavioral therapy" project that has been developed and implemented as part of a federal re-entry court. Finally, we discuss the implications of the challenges of intervention at this stage, and the recently developed "Re-entry Project," for research, policy, and practice. Copyright © 2017 John Wiley & Sons, Ltd.

Re-entry Adjustment and Job Embeddedness: The Mediating Role of Professional Identity in Indonesian Returnees.

PubMed

Andrianto, Sonny; Jianhong, Ma; Hommey, Confidence; Damayanti, Devi; Wahyuni, Honey

2018-01-01

The present study examined the relationship between difficulty in re-entry adjustment and job embeddedness, considering the mediating role of sense of professional identity. The online data on demographic characteristics, difficulty on re-entry adjustment, sense of professional identity, and job embeddedness were collected from 178 Indonesian returnees from multiple organizations. The results showed that difficulty in re-entry adjustment was a significant predictor of a sense of professional identity; a sense of professional identity was a significant predictor of job embeddedness. Furthermore, sense of professional identity is an effective mediating variable, bridging the relationship between post-return conditions to the home country and work atmosphere. Finally, the key finding of this study was that sense of professional identity mediated the effect of difficulty in re-entry adjustment on job embeddedness. The theoretical and practical implications, study limitations, and future research needs of our findings are noted.

78 FR 66898 - Low Enriched Uranium From France: Final Results of Changed Circumstances Review

Federal Register 2010, 2011, 2012, 2013, 2014

2013-11-07

... in U.S. customs territory, and (ii) are re-exported within eighteen (18) months of entry of the low... extend the deadline for re-exportation of this sole entry of low-enriched uranium. The Department determines that the deadline for re-exportation of this sole entry is November 1, 2015, and that this will be...

State Policies Affecting the "Adult Re-Entry Pipeline" in Postsecondary Education: Results of a Fifty-State Inventory

ERIC Educational Resources Information Center

Boeke, Marianne; Zis, Stacey; Ewell, Peter

2011-01-01

With support from the Bill and Melinda Gates Foundation, the National Center for Higher Education Management Systems (NCHEMS) is engaged in a two year project centered on state policies that foster student progression and success in the "adult re-entry pipeline." The adult re-entry pipeline consists of the many alternative pathways to…

HIV-1 requires Arf6-mediated membrane dynamics to efficiently enter and infect T lymphocytes

PubMed Central

García-Expósito, Laura; Barroso-González, Jonathan; Puigdomènech, Isabel; Machado, José-David; Blanco, Julià; Valenzuela-Fernández, Agustín

2011-01-01

As the initial barrier to viral entry, the plasma membrane along with the membrane trafficking machinery and cytoskeleton are of fundamental importance in the viral cycle. However, little is known about the contribution of plasma membrane dynamics during early human immunodeficiency virus type 1 (HIV-1) infection. Considering that ADP ribosylation factor 6 (Arf6) regulates cellular invasion via several microorganisms by coordinating membrane trafficking, our aim was to study the function of Arf6-mediated membrane dynamics on HIV-1 entry and infection of T lymphocytes. We observed that an alteration of the Arf6–guanosine 5′-diphosphate/guanosine 5′-triphosphate (GTP/GDP) cycle, by GDP-bound or GTP-bound inactive mutants or by specific Arf6 silencing, inhibited HIV-1 envelope–induced membrane fusion, entry, and infection of T lymphocytes and permissive cells, regardless of viral tropism. Furthermore, cell-to-cell HIV-1 transmission of primary human CD4+ T lymphocytes was inhibited by Arf6 knockdown. Total internal reflection fluorescence microscopy showed that Arf6 mutants provoked the accumulation of phosphatidylinositol-(4,5)-biphosphate–associated structures on the plasma membrane of permissive cells, without affecting CD4-viral attachment but impeding CD4-dependent HIV-1 entry. Arf6 silencing or its mutants did not affect fusion, entry, and infection of vesicular stomatitis virus G–pseudotyped viruses or ligand-induced CXCR4 or CCR5 endocytosis, both clathrin-dependent processes. Therefore we propose that efficient early HIV-1 infection of CD4+ T lymphocytes requires Arf6-coordinated plasma membrane dynamics that promote viral fusion and entry. PMID:21346189

Hepatitis C virus depends on E-cadherin as an entry factor and regulates its expression in epithelial-to-mesenchymal transition.

PubMed

Li, Qisheng; Sodroski, Catherine; Lowey, Brianna; Schweitzer, Cameron J; Cha, Helen; Zhang, Fang; Liang, T Jake

2016-07-05

Hepatitis C virus (HCV) enters the host cell through interactions with a cascade of cellular factors. Although significant progress has been made in understanding HCV entry, the precise mechanisms by which HCV exploits the receptor complex and host machinery to enter the cell remain unclear. This intricate process of viral entry likely depends on additional yet-to-be-defined cellular molecules. Recently, by applying integrative functional genomics approaches, we identified and interrogated distinct sets of host dependencies in the complete HCV life cycle. Viral entry assays using HCV pseudoparticles (HCVpps) of various genotypes uncovered multiple previously unappreciated host factors, including E-cadherin, that mediate HCV entry. E-cadherin silencing significantly inhibited HCV infection in Huh7.5.1 cells, HepG2/miR122/CD81 cells, and primary human hepatocytes at a postbinding entry step. Knockdown of E-cadherin, however, had no effect on HCV RNA replication or internal ribosomal entry site (IRES)-mediated translation. In addition, an E-cadherin monoclonal antibody effectively blocked HCV entry and infection in hepatocytes. Mechanistic studies demonstrated that E-cadherin is closely associated with claudin-1 (CLDN1) and occludin (OCLN) on the cell membrane. Depletion of E-cadherin drastically diminished the cell-surface distribution of these two tight junction proteins in various hepatic cell lines, indicating that E-cadherin plays an important regulatory role in CLDN1/OCLN localization on the cell surface. Furthermore, loss of E-cadherin expression in hepatocytes is associated with HCV-induced epithelial-to-mesenchymal transition (EMT), providing an important link between HCV infection and liver cancer. Our data indicate that a dynamic interplay among E-cadherin, tight junctions, and EMT exists and mediates an important function in HCV entry.

Re-Entry Point Targeting for LEO Spacecraft using Aerodynamic Drag

NASA Technical Reports Server (NTRS)

Omar, Sanny; Bevilacqua, Riccardo; Fineberg, Laurence; Treptow, Justin; Johnson, Yusef; Clark, Scott

2016-01-01

Most Low Earth Orbit (LEO) spacecraft do not have thrusters and re-enter atmosphere in random locations at uncertain times. Objects pose a risk to persons, property, or other satellites. Has become a larger concern with the recent increase in small satellites. Working on a NASA funded project to design a retractable drag device to expedite de-orbit and target a re-entry location through modulation of the drag area. Will be discussing the re-entry point targeting algorithm here.

Coordinating cell proliferation and differentiation: Antagonism between cell cycle regulators and cell type-specific gene expression

PubMed Central

Ruijtenberg, Suzan; van den Heuvel, Sander

2016-01-01

ABSTRACT Cell proliferation and differentiation show a remarkable inverse relationship. Precursor cells continue division before acquiring a fully differentiated state, while terminal differentiation usually coincides with proliferation arrest and permanent exit from the division cycle. Mechanistic insight in the temporal coordination between cell cycle exit and differentiation has come from studies of cells in culture and genetic animal models. As initially described for skeletal muscle differentiation, temporal coordination involves mutual antagonism between cyclin-dependent kinases that promote cell cycle entry and transcription factors that induce tissue-specific gene expression. Recent insights highlight the contribution of chromatin-regulating complexes that act in conjunction with the transcription factors and determine their activity. In particular SWI/SNF chromatin remodelers contribute to dual regulation of cell cycle and tissue-specific gene expression during terminal differentiation. We review the concerted regulation of the cell cycle and cell type-specific transcription, and discuss common mutations in human cancer that emphasize the clinical importance of proliferation versus differentiation control. PMID:26825227

77 FR 19642 - Low Enriched Uranium From France: Final Results of Antidumping Duty Changed Circumstances Review

Federal Register 2010, 2011, 2012, 2013, 2014

2012-04-02

... in U.S. customs territory, and (ii) are re-exported within eighteen (18) months of entry of the LEU... amend the scope of the order and to extend the deadline for the re-exportation of this sole LEU entry... transporter(s) while in U.S. customs territory, and (ii) are re-exported within eighteen (18) months of entry...

The Rewarding Challenge: Welcoming Re-Entry Women Students to the Small College.

ERIC Educational Resources Information Center

Midland Lutheran Coll., Fremont, NE.

This handbook was designed to offer a low-cost, local, and practical approach to working with re-entry women college students. It is intended for use by staff in small colleges (under 1,000 students) where women's centers and on-campus child care are rare and where the needs of re-entry students have not been adequately addressed. The information…

Cell cycle entry triggers a switch between two modes of Cdc42 activation during yeast polarization

PubMed Central

Witte, Kristen; Strickland, Devin; Glotzer, Michael

2017-01-01

Cell polarization underlies many cellular and organismal functions. The GTPase Cdc42 orchestrates polarization in many contexts. In budding yeast, polarization is associated with a focus of Cdc42•GTP which is thought to self sustain by recruiting a complex containing Cla4, a Cdc42-binding effector, Bem1, a scaffold, and Cdc24, a Cdc42 GEF. Using optogenetics, we probe yeast polarization and find that local recruitment of Cdc24 or Bem1 is sufficient to induce polarization by triggering self-sustaining Cdc42 activity. However, the response to these perturbations depends on the recruited molecule, the cell cycle stage, and existing polarization sites. Before cell cycle entry, recruitment of Cdc24, but not Bem1, induces a metastable pool of Cdc42 that is sustained by positive feedback. Upon Cdk1 activation, recruitment of either Cdc24 or Bem1 creates a stable site of polarization that induces budding and inhibits formation of competing sites. Local perturbations have therefore revealed unexpected features of polarity establishment. DOI: http://dx.doi.org/10.7554/eLife.26722.001 PMID:28682236

Genome-wide expression profiling of in vivo-derived bloodstream parasite stages and dynamic analysis of mRNA alterations during synchronous differentiation in Trypanosoma brucei

PubMed Central

Kabani, Sarah; Fenn, Katelyn; Ross, Alan; Ivens, Al; Smith, Terry K; Ghazal, Peter; Matthews, Keith

2009-01-01

Background Trypanosomes undergo extensive developmental changes during their complex life cycle. Crucial among these is the transition between slender and stumpy bloodstream forms and, thereafter, the differentiation from stumpy to tsetse-midgut procyclic forms. These developmental events are highly regulated, temporally reproducible and accompanied by expression changes mediated almost exclusively at the post-transcriptional level. Results In this study we have examined, by whole-genome microarray analysis, the mRNA abundance of genes in slender and stumpy forms of T.brucei AnTat1.1 cells, and also during their synchronous differentiation to procyclic forms. In total, five biological replicates representing the differentiation of matched parasite populations derived from five individual mouse infections were assayed, with RNAs being derived at key biological time points during the time course of their synchronous differentiation to procyclic forms. Importantly, the biological context of these mRNA profiles was established by assaying the coincident cellular events in each population (surface antigen exchange, morphological restructuring, cell cycle re-entry), thereby linking the observed gene expression changes to the well-established framework of trypanosome differentiation. Conclusion Using stringent statistical analysis and validation of the derived profiles against experimentally-predicted gene expression and phenotypic changes, we have established the profile of regulated gene expression during these important life-cycle transitions. The highly synchronous nature of differentiation between stumpy and procyclic forms also means that these studies of mRNA profiles are directly relevant to the changes in mRNA abundance within individual cells during this well-characterised developmental transition. PMID:19747379

Regulation of Mih1/Cdc25 by protein phosphatase 2A and casein kinase 1

PubMed Central

Pal, Gayatri; Paraz, Maria T.Z.; Kellogg, Douglas R.

2008-01-01

The Cdc25 phosphatase promotes entry into mitosis by removing cyclin-dependent kinase 1 (Cdk1) inhibitory phosphorylation. Previous work suggested that Cdc25 is activated by Cdk1 in a positive feedback loop promoting entry into mitosis; however, it has remained unclear how the feedback loop is initiated. To learn more about the mechanisms that regulate entry into mitosis, we have characterized the function and regulation of Mih1, the budding yeast homologue of Cdc25. We found that Mih1 is hyperphosphorylated early in the cell cycle and is dephosphorylated as cells enter mitosis. Casein kinase 1 is responsible for most of the hyperphosphorylation of Mih1, whereas protein phosphatase 2A associated with Cdc55 dephosphorylates Mih1. Cdk1 appears to directly phosphorylate Mih1 and is required for initiation of Mih1 dephosphorylation as cells enter mitosis. Collectively, these observations suggest that Mih1 regulation is achieved by a balance of opposing kinase and phosphatase activities. Because casein kinase 1 is associated with sites of polar growth, it may regulate Mih1 as part of a signaling mechanism that links successful completion of growth-related events to cell cycle progression. PMID:18316413

The X-38 V-201 Flap Actuator Mechanism

NASA Technical Reports Server (NTRS)

Hagen, Jeff; Moore, Landon; Estes, Jay; Layer, Chris

2004-01-01

The X-38 Crew Rescue Vehicle V-201 space flight test article was designed to achieve an aerodynamically controlled re-entry from orbit in part through the use of two body mounted flaps on the lower rear side. These flaps are actuated by an electromechanical system that is partially exposed to the re-entry environment. These actuators are of a novel configuration and are unique in their requirement to function while exposed to re-entry conditions. The authors are not aware of any other vehicle in which a major actuator system was required to function throughout the complete re-entry profile while parts of the actuator were directly exposed to the ambient environment.

Rotaviruses induce an early membrane permeabilization of MA104 cells and do not require a low intracellular Ca2+ concentration to initiate their replication cycle.

PubMed Central

Cuadras, M A; Arias, C F; López, S

1997-01-01

In this work, we found that rotavirus infection induces an early membrane permeabilization of MA104 cells and promotes the coentry of toxins, such as alpha-sarcin, into the cell. This cell permeability was shown to depend on infectious virus and was also shown to be virus dose dependent, with 10 infectious particles per cell being sufficient to achieve maximum permeability; transient, lasting no more than 15 min after virus entry and probably occurring concomitantly with virus penetration; and specific, since cells that are poorly permissive for rotavirus were not permeabilized. The rotavirus-mediated coentry of toxins was not blocked by the endocytosis inhibitors dansylcadaverine and cytochalasin D or by the vacuolar proton-ATPase inhibitor bafilomycin A1, suggesting that neither endocytocis nor an intraendosomal acidic pH or a proton gradient is required for permeabilization of the cells. Compounds that raise the intracellular concentration of calcium ([Ca2+]i) by different mechanisms, such as the calcium ionophores A23187 and ionomycin and the endoplasmic reticulum calcium-ATPase inhibitor thapsigargin, did not block the coentry of alpha-sarcin or affect the onset of viral protein synthesis, suggesting that a low [Ca2+]i is not essential for the initial steps of the virus life cycle. Since the entry of alpha-sarcin correlates with virus penetration in all parameters tested, the assay for permeabilization to toxins might be a useful tool for studying and characterizing the route of entry and the mechanism used by rotaviruses to traverse the cell membrane and initiate a productive replication cycle. PMID:9371563

Trajectory Design and Control for the Compton Gamma Ray Observatory Re-Entry

NASA Technical Reports Server (NTRS)

Hoge, Susan; Vaughn, Frank J., Jr.

2001-01-01

The Compton Gamma Ray Observatory (CGRO) controlled re-entry operation was successfully conducted in June of 2000. The surviving parts of the spacecraft landed in the Pacific Ocean within the nominal impact target zone. The design of the maneuvers to control the trajectory to accomplish this re-entry presented several challenges. These challenges included the timing and duration of the maneuvers, propellant management, post-maneuver state determination, collision avoidance with other spacecraft, accounting for the break-up of the spacecraft into several pieces with a wide range of ballistic coefficients, and ensuring that the impact footprint would remain within the desired impact target zone in the event of contingencies. This paper presents the initial re-entry trajectory design and traces the evolution of that design into the maneuver sequence used for the re-entry. The paper also discusses the spacecraft systems and operational constraints imposed on the trajectory design and the required modifications to the initial design based on those constraints. Data from the reentry operation are also presented.

Nuclear receptor TLX prevents retinal dystrophy and recruits the corepressor atrophin1

PubMed Central

Zhang, Chun-Li; Zou, Yuhua; Yu, Ruth T.; Gage, Fred H.; Evans, Ronald M.

2006-01-01

During mammalian embryogenesis, precise coordination of progenitor cell proliferation and differentiation is essential for proper organ size and function. The involvement of TLX (NR2E1), an orphan nuclear receptor, has been implicated in ocular development, as Tlx−/− mice exhibit visual impairment. Using genetic and biochemical approaches, we show that TLX modulates retinal progenitor cell proliferation and cell cycle re-entry by directly regulating the expression of Pten and its target cyclin D1. Additionally, TLX finely tunes the progenitor differentiation program by modulating the phospholipase C and mitogen-activated protein kinase (MAPK) pathways and the expression of an array of cell type-specific transcriptional regulators. Consequently, Tlx−/− mice have a dramatic reduction in retina thickness and enhanced generation of S-cones, and develop severe early onset retinal dystrophy. Furthermore, TLX interacts with atrophin1 (Atn1), a corepressor that is involved in human neurodegenerative dentatorubral-pallidoluysian atrophy (DRPLA) and that is essential for development of multiple tissues. Together, these results reveal a molecular strategy by which an orphan nuclear receptor can precisely orchestrate tissue-specific proliferation and differentiation programs to prevent retinal malformation and degeneration. PMID:16702404

A Novel 'Cheese Wire' Technique for Stent Positioning Following Difficult Iliac Artery Subintimal Dissection and Aortic Re-Entry

DOE Office of Scientific and Technical Information (OSTI.GOV)

Watkinson, A. F., E-mail: anthony.watkinson@rdeft.nhs.u

2009-07-15

Subintimal wire dissection is a well-established method for traversing difficult vascular occlusions. This technique relies on re-entry of the true lumen distal to the occlusion, which may be difficult in diseased vessels with significant calcification. This case report describes a novel 'cheese wire' technique to allow stent positioning without the use of proprietary re-entry devices.

Impact Testing of the H1224A Shipping/Storage Container

DTIC Science & Technology

1994-05-01

may not provide significant ener- gy absorption for the re - entry vehicle midsection but can provide some confinement of potentially damaged...Horizontal Low-Velocity impact test LHV Longitudinal High-Velocity impact test HHV Horizontal High-Velocity impact test RV Re - entry Vehicle midsection mass...Also, integration of these pulses showed that only a much shorter dura- tion pulse was necessary to slow the re - entry vehicle midsection velocity

Initial experience with a dedicated coronary re-entry device for revascularization of chronic total occlusions.

PubMed

Whitlow, Patrick L; Lombardi, William L; Araya, Mario; Michael Wyman, R; Torres, Humberto; Dauvergne, Christian; Tsuchikane, Etsuo; Lansky, Alexandra; Thompson, Craig A

2012-11-01

The aim of this registry was to evaluate a new device designed to facilitate antegrade guidewire re-entry into the true lumen of a chronic total coronary occlusion (CTO) from the adjacent subintimal space. Successful recanalization of CTOs results in clinical improvement in appropriately selected patients. CTO intervention is time- and resource-consuming, and a simplified approach enabling antegrade guidewire re-entry into the distal true lumen might improve success. Patients with CTO and ischemia were entered into a prospective registry regardless of lesion characteristics. If wire manipulation resulted in subintimal wire entrapment, a new re-entry tool (a 2.5-mm flat subintimal balloon with two exit ports offset by 180°) was used as a platform to attempt guidewire penetration into the distal true lumen. The primary endpoint assessed was successful device-guided re-entry. Standard techniques were then utilized to open the CTO. In 40 consecutive CTO lesions attempted, 19 resulted in subintimal wire entrapment (mean occlusion length 44 mm). Sixteen of these 19 were successfully crossed with an antegrade guidewire into the distal true lumen using the new device (84%). One patient with unsuccessful re-entry was subsequently recanalized with a retrograde technique. All crossed lesions were stented (17/17), resulting in TIMI 3 flow without major complications. Two cases were unsuccessful. One patient had a grade I coronary perforation requiring no treatment. A new device to recanalize CTOs complicated by subintimal wire entrapment can be used successfully by experienced operators. Further study of this coronary re-entry device is ongoing. Copyright © 2011 Wiley Periodicals, Inc.

How do children with a chronic or long-term illness perceive their school re-entry after a period of homebound instruction?

PubMed

Boonen, H; Petry, K

2012-07-01

A considerable number of children are confronted with a chronic or long-term illness in their lives. For these children, absenteeism is problematic, because education plays a major role in stimulating their cognitive development and in promoting a sense of normalcy and psychosocial well-being. In the literature, a great deal of attention has been paid to school reintegration programmes, which try to counter the barriers that these children may face when they return to school. Another way of surmounting these barriers is through the use of homebound instruction, in which the educational process for the child is continued during the period of absence. Despite the growing awareness of the necessity of education for these children, there is still little empirical research available addressing programmes that facilitate school re-entry. The major goal of this study is to investigate how parents and their children with a chronic or long-term illness perceive school re-entry after a period of homebound instruction, by using a descriptive-explorative, multi-informant research design. Participants were 60 children and their parents who filled in a self-constructed questionnaire. Both parents and children perceived the period of homebound instruction, as well as their school re-entry, predominantly positively. Most of the children stated that they had been able to keep up with their subjects, and that they had good contact with their peers when they returned to school. According to parents, homebound instruction made a positive contribution to the school re-entry of their child. The current study is one of the first to explore the school re-entry of children with a chronic or long-term illness. According to both parents and children, the school re-entry process passed off positively. However, more research is needed with regard to the quality of education and the programmes aimed at facilitating school re-entry. © 2011 Blackwell Publishing Ltd.

MS Garneau in his LES during re-entry preparations for STS-97

NASA Image and Video Library

2000-12-11

STS097-310-026 (11 December 2000) --- Astronaut Marc Garneau, mission specialist representing the Canadian Space Agency (CSA), is photographed in the launch and entry suit on the middeck of the Earth-orbiting Space Shuttle Endeavour prior to re-entry.

Novel Insights into Cell Entry of Emerging Human Pathogenic Arenaviruses.

PubMed

Fedeli, Chiara; Moreno, Héctor; Kunz, Stefan

2018-06-22

Viral hemorrhagic fevers caused by emerging RNA viruses of the Arenavirus family are among the most devastating human diseases. Climate change, global trade, and increasing urbanization promote the emergence and re-emergence of these human pathogenic viruses. Emerging pathogenic arenaviruses are of zoonotic origin and reservoir-to-human transmission is crucial for spillover into human populations. Host cell attachment and entry are the first and most fundamental steps of every virus infection and represent major barriers for zoonotic transmission. During host cell invasion, viruses critically depend on cellular factors, including receptors, co-receptors, and regulatory proteins of endocytosis. An in-depth understanding of the complex interaction of a virus with cellular factors implicated in host cell entry is therefore crucial to predict the risk of zoonotic transmission, define the tissue tropism, and assess disease potential. Over the past years, investigation of the molecular and cellular mechanisms underlying host cell invasion of human pathogenic arenaviruses uncovered remarkable viral strategies and provided novel insights into viral adaptation and virus-host co-evolution that will be covered in the present review. Copyright © 2018. Published by Elsevier Ltd.

Airborne Observation of the Hayabusa Sample Return Capsule Re-Entry

NASA Technical Reports Server (NTRS)

Grinstead, Jay H.; Jenniskens, Peter; Cassell, Alan M.; Albers, James; Winter, Michael W.

2011-01-01

NASA Ames Research Center and the SETI Institute collaborated on an effort to observe the Earth re-entry of the Japan Aerospace Exploration Agency's Hayabusa sample return capsule. Hayabusa was an asteroid exploration mission that retrieved a sample from the near-Earth asteroid Itokawa. Its sample return capsule re-entered over the Woomera Prohibited Area in southern Australia on June 13, 2010. Being only the third sample return mission following NASA's Genesis and Stardust missions, Hayabusa's return was a rare opportunity to collect aerothermal data from an atmospheric entry capsule returning at superorbital speeds. NASA deployed its DC-8 airborne laboratory and a team of international researchers to Australia for the re-entry. For approximately 70 seconds, spectroscopic and radiometric imaging instruments acquired images and spectra of the capsule, its wake, and destructive re-entry of the spacecraft bus. Once calibrated, spectra of the capsule will be interpreted to yield data for comparison with and validation of high fidelity and engineering simulation tools used for design and development of future atmospheric entry system technologies. A brief summary of the Hayabusa mission, the preflight preparations and observation mission planning, mission execution, and preliminary spectral data are documented.

Tetraploidization or autophagy: The ultimate fate of senescent human endometrial stem cells under ATM or p53 inhibition.

PubMed

Borodkina, Aleksandra V; Shatrova, Alla N; Deryabin, Pavel I; Grukova, Anastasiya A; Nikolsky, Nikolay N; Burova, Elena B

2016-01-01

Previously we demonstrated that endometrium-derived human mesenchymal stem cells (hMESCs) via activation of the ATM/p53/p21/Rb pathway enter the premature senescence in response to oxidative stress. Down regulation effects of the key components of this signaling pathway, particularly ATM and p53, on a fate of stressed hMESCs have not yet been investigated. In the present study by using the specific inhibitors Ku55933 and Pifithrin-α, we confirmed implication of both ATM and p53 in H(2)O(2)-induced senescence of hMESCs. ATM or p53 down regulation was shown to modulate differently the cellular fate of H(2)O(2)-treated hMESCs. ATM inhibition allowed H(2)O(2)-stimulated hMESCs to escape the permanent cell cycle arrest due to loss of the functional ATM/p53/p21/Rb pathway, and induced bypass of mitosis and re-entry into S phase, resulting in tetraploid cells. On the contrary, suppression of the p53 transcriptional activity caused a pronounced cell death of H(2)O(2)-treated hMESCs via autophagy induction. The obtained data clearly demonstrate that down regulation of ATM or p53 shifts senescence of human endometrial stem cells toward tetraploidization or autophagy.

Radiant Heat Testing of the H1224A Shipping/Storage Container

DTIC Science & Technology

1994-05-01

re - entry vehicles caused by credible accidents during air and ground transportation. Radiant heat testing of the H1224A storage/shipping container is...inner container, and re - entry vehicle (RV) temperatures during radiant heat testing. Computer modelling can be used to predict weapon response throughout...Nomenclature RV Re - entry Vehicle midsection mass mock-up WR War Reserve STS Stockpile-to-Target Sequence NAWC Simulated H1224A container by Naval Air

Interaction of Human Tumor Viruses with Host Cell Surface Receptors and Cell Entry

PubMed Central

Schäfer, Georgia; Blumenthal, Melissa J.; Katz, Arieh A.

2015-01-01

Currently, seven viruses, namely Epstein-Barr virus (EBV), Kaposi’s sarcoma-associated herpes virus (KSHV), high-risk human papillomaviruses (HPVs), Merkel cell polyomavirus (MCPyV), hepatitis B virus (HBV), hepatitis C virus (HCV) and human T cell lymphotropic virus type 1 (HTLV-1), have been described to be consistently associated with different types of human cancer. These oncogenic viruses belong to distinct viral families, display diverse cell tropism and cause different malignancies. A key to their pathogenicity is attachment to the host cell and entry in order to replicate and complete their life cycle. Interaction with the host cell during viral entry is characterized by a sequence of events, involving viral envelope and/or capsid molecules as well as cellular entry factors that are critical in target cell recognition, thereby determining cell tropism. Most oncogenic viruses initially attach to cell surface heparan sulfate proteoglycans, followed by conformational change and transfer of the viral particle to secondary high-affinity cell- and virus-specific receptors. This review summarizes the current knowledge of the host cell surface factors and molecular mechanisms underlying oncogenic virus binding and uptake by their cognate host cell(s) with the aim to provide a concise overview of potential target molecules for prevention and/or treatment of oncogenic virus infection. PMID:26008702

Distal re-entry closure with neobranching technique after thoracic endovascular aortic repair of Type B aortic dissection.

PubMed

Yamamoto, Masaki; Fukutomi, Takashi; Noguchi, Tatsuya; Orihashi, Kazumasa

2018-04-01

Retrograde false-lumen flow after thoracic endovascular aortic repair of Type B aortic dissection occurs occasionally and may have a negative impact on aortic remodelling and even prevent the decompression of the false lumen. A 67-year-old man with a Type B aortic dissection underwent thoracic endovascular aortic repair for severe compression of the true lumen and visceral malperfusion 7 weeks after the onset. Intraoperative angiography revealed proximal entry tear closure, but the false-lumen flow increased because of retrograde flow through the re-entry tear. Additional intervention including re-entry tear closure was performed with a neobranching technique with covered stent placement in the visceral artery from the aortic true lumen through the distal re-entry tear. We report a case of Type B aortic dissection and discuss the surgical techniques used.

Parasystole due to re-entry as the possible mechanism of ventricular parasystole with second-degree entrance block.

PubMed

Kinoshita, Shinji; Katoh, Takakazu; Yoshida, Hiroshi

2010-05-01

In 1974, Kinoshita reported a case of 'irregular parasystole' due to type I second-degree entrance block. Since then, many cases of such 'irregular' parasystole have been reported by us. To explain the mechanism of 'irregular' parasystole, two theories have been suggested, namely, 'electrotonic modulation' by Jalife and Moe, and 'type I second-degree entrance block' by us. On the contrary, in 1960, Kinoshita et al. reported a case of concealed bigeminy for the first time. The electrocardiographic findings in concealed bigeminy have suggested that there are dual re-entrant pathways with markedly long effective refractory periods in the re-entrant pathway. We have suggested that parasystole may be caused by re-entry in such re-entrant pathways. In this article, attempts are made to explain the mechanism of all the electrocardiographic findings in our cases of parasystole by 'parasystole due to re-entry'. Using 24 studies on parasystole and 21 studies on concealed extrasystoles that we have reported over 50 years, as well as three exemplary cases in this article, attempts are made to explain all electrocardiographic findings in parasystole by 'parasystole due to re-entry'. The electrocardiographic findings in our previous clinical cases of parasystole and concealed extrasystoles, as well as exemplary cases and diagrams in the present article, strongly suggest 'parasystole due to re-entry' as the mechanism of ventricular parasystole with second-degree entrance block.

Positive Feedback Keeps Duration of Mitosis Temporally Insulated from Upstream Cell-Cycle Events.

PubMed

Araujo, Ana Rita; Gelens, Lendert; Sheriff, Rahuman S M; Santos, Silvia D M

2016-10-20

Cell division is characterized by a sequence of events by which a cell gives rise to two daughter cells. Quantitative measurements of cell-cycle dynamics in single cells showed that despite variability in G1-, S-, and G2 phases, duration of mitosis is short and remarkably constant. Surprisingly, there is no correlation between cell-cycle length and mitotic duration, suggesting that mitosis is temporally insulated from variability in earlier cell-cycle phases. By combining live cell imaging and computational modeling, we showed that positive feedback is the molecular mechanism underlying the temporal insulation of mitosis. Perturbing positive feedback gave rise to a sluggish, variable entry and progression through mitosis and uncoupled duration of mitosis from variability in cell cycle length. We show that positive feedback is important to keep mitosis short, constant, and temporally insulated and anticipate it might be a commonly used regulatory strategy to create modularity in other biological systems. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

Short-Lasting Episodes of Torsade de Pointes in the Chronic Atrioventricular Block Dog Model Have a Focal Mechanism, While Longer-Lasting Episodes Are Maintained by Re-Entry.

PubMed

Vandersickel, Nele; Bossu, Alexandre; De Neve, Jan; Dunnink, Albert; Meijborg, Veronique M F; van der Heyden, Marcel A G; Beekman, Jet D M; De Bakker, Jacques M T; Vos, Marc A; Panfilov, Alexander V

2017-12-26

This study investigated the arrhythmogenic mechanisms responsible for torsade de pointes (TdP) in the chronic atrioventricular block dog model, known for its high susceptibility for TdP. The mechanism of TdP arrhythmias has been under debate for many years. Focal activity as well as re-entry have both been mentioned in the initiation and the perpetuation of TdP. In 5 TdP-sensitive chronic atrioventricular block dogs, 56 needle electrodes were evenly distributed transmurally to record 240 unipolar local electrograms simultaneously. Nonterminating (NT) episodes were defibrillated after 10 s. Software was developed to automatically detect activation times and to create 3-dimensional visualizations of the arrhythmia. For each episode of ectopic activity (ranging from 2 beats to NT episodes), a novel methodology was created to construct directed graphs of the wave propagation and detect re-entry loops by using an iterative depth-first-search algorithm. Depending on the TdP definition (number of consecutive ectopic beats), we analyzed 29 to 54 TdP: 29 were longer than 5 beats. In the total group, 9 were NT and 45 were self-terminating. Initiation and termination were always based on focal activity. Re-entry becomes more important in the longer-lasting episodes (>14 beats), whereas in all NT TdP, re-entry was the last active mechanism. During re-entry, excitation fronts were constantly present in the heart, while during focal TdP, there was always a silent interval between 2 consecutive waves (142 ms) during which excitation fronts were absent. Interbeat intervals were significantly smaller for re-entry episodes-220 versus 310 ms in focal. Electrograms recorded in particular areas during NT TdP episodes had significantly smaller amplitude (0.38) than during focal episodes (0.59). TdP can be driven by focal activity as well as by re-entry depending on the duration of the episode. NT episodes are always maintained by re-entry, which can be identified in local unipolar electrograms by shorter interbeat intervals and smaller deflection amplitude. Copyright © 2017 American College of Cardiology Foundation. Published by Elsevier Inc. All rights reserved.

FLPP IXV Re-Entry Vehicle, Supersonic Charectisation Based on DNW SST Wind Tunnel Tests and CFD

NASA Astrophysics Data System (ADS)

Kapteijn, C.; Maseland, H.; Chiarelli, C.; Mareschi, V.; Tribot, J.-P.; Binetti, P.; Walloscheck, T.

2009-01-01

The European Space Agency ESA, has engaged in 2004, the IXV project (Intermediate eXperimental Vehicle) which is part of the FLPP (Future Launcher Preparatory Programme) aiming at answering to critical technological issues for controlled re-entry, while supporting the future generation launchers and to improve in general European capabilities in the strategic field of atmospheric re-entry for future space transportation, exploration and scientific applications. The IXV key mission and system objectives are the design, development, manufacturing, assembling and on- ground to in-flight verification of an autonomous European lifting and aerodynamically controlled re- entry system, integrating the critical re- entry technologies at the system level. In particular, the IXV shall demonstrate system integrated key technologies such as lifting flight control by means of aerodynamic surfaces that are one of the main primary objectives of the experimental investigation. Lifting and aerodynamic controlled re-entry represents a significant capability advancement with respect to the ballistic re-entry of capsules like the ARD. Since hypersonic aerodynamics is essentially different from supersonic aerodynamics, the current mission is to perform an atmospheric re-entry in combination with a safe recovery the in supersonic flight regime. However, mission extension to trimmed transonic flight is under consideration based on a preliminary analysis of the aerodynamic characteristics of the IXV configuration. Since the beginning of the IXV project, an aerodynamic data base (AEDB) has been built up and continuously updated integrating the additional information mainly provided by means of CFD (ie: Euler and Navier-Stokes) and lately also by means of WTTs. This AEDB serves for flying qualities analysis and for re-entry simulations. During the development phase B2/C1, the effectiveness of the control surfaces and their impact on te vehicle's aerodynamic forces in the supersonic regime is measured for a number of discrete deflection settings in the Super-Sonic wind Tunnel (SST) of DNW. Enabling an improved understanding of the measured aerodynamic characteristics, complementary computations were performed by Thales Alenia Space. The complete set of data was analyzed and compared enabling a consolidation of the nominal aerodynamic and aerodynamic uncertainties as well. The paper presents the main objectives of the supersonic characterisation of IXV including WTTs, and the main outcomes of the current data comparisons.

Regulation of store-operated Ca2+ entry activity by cell cycle dependent up-regulation of Orai2 in brain capillary endothelial cells.

PubMed

Kito, Hiroaki; Yamamura, Hisao; Suzuki, Yoshiaki; Yamamura, Hideto; Ohya, Susumu; Asai, Kiyofumi; Imaizumi, Yuji

2015-04-10

Store-operated Ca(2+) entry (SOCE) via Orai1 and STIM1 complex is supposed to have obligatory roles in the regulation of cellular functions of vascular endothelial cells, while little is known about the contribution of Orai2. Quantitative PCR and Western blot analyses indicated the expression of Orai2 and STIM2, in addition to Orai1 and STIM1 in bovine brain capillary endothelial cell line, t-BBEC117. During the exponential growth of t-BBEC117, the knockdown of Orai1 and STIM1 significantly reduced the SOCE activity, whereas Orai2 and STIM2 siRNAs had no effect. To examine whether endogenous SOCE activity contributes to the regulation of cell cycle progression, t-BBEC117 were synchronized using double thymidine blockage. At the G2/M phase, Ca(2+) influx via SOCE was decreased and Orai2 expression was increased compared to the G0/G1 phase. When Orai2 was knocked down at the G2/M phase, the decrease in SOCE was removed, and cell proliferation was partly attenuated. Taken together, Orai1 significantly contributes to cell proliferation via the functional expression, which is presumably independent of the cell cycle phases. In construct, Orai2 is specifically up-regulated during the G2/M phase, negatively modulates the SOCE activity, and may contribute to the regulation of cell cycle progression in brain capillary endothelial cells. Copyright © 2015 Elsevier Inc. All rights reserved.

Levels and determinants of pesticide exposure in re-entry workers in vineyards: results of the PESTEXPO study.

PubMed

Baldi, Isabelle; Lebailly, P; Bouvier, G; Rondeau, V; Kientz-Bouchart, V; Canal-Raffin, M; Garrigou, A

2014-07-01

Physical contact with branches, leaves, fruit or vegetables in previously treated crops is responsible for the transfer of pesticides to the worker's skin in agricultural tasks such as harvesting, pruning, thinning, cutting or sorting. Few studies have documented workers' exposure during re-entry in vineyards. In the PESTEXPO study, we described levels of exposure and analyzed their determinants during re-entry and harvesting in vineyards in the Bordeaux area, France. Between 2002 and 2007, volunteers performing re-entry tasks (N=46 days) or harvesting (N=48 days) after dithiocarbamate or folpet treatment were observed. Detailed information on the tasks was collected and dermal contamination was assessed using patches placed on the skin and hand-washing at the end of each working phase. Daily median contamination was 1 967.7 μl of mixture during re-entry (90(e) percentile: 5 045.3 μl) and 18.7 μl during harvesting (90(e) percentile: 911.4 μl). The type of task was the parameter found to be the most strongly associated with contamination. For re-entry, the highest contaminations were observed during raising of wires and cutting of branches. During the harvest, the contamination was maximal for grape-picking. The delay since the last treatment and the rate of active ingredient per hectare played a role, together with other factors such as meteorological factors, crop and farm characteristics, gloves and clothes. Our results underline the necessity to take into account exposures during re-entry and harvest when considering pesticide exposure, both for epidemiological research and preventive action. Copyright © 2014 Elsevier Inc. All rights reserved.

Measles Virus Enters Breast and Colon Cancer Cell Lines through a PVRL4-Mediated Macropinocytosis Pathway

PubMed Central

Delpeut, Sebastien; Sisson, Gary; Black, Karen M.

2017-01-01

ABSTRACT Measles virus (MeV) is a member of the family Paramixoviridae that causes a highly contagious respiratory disease but has emerged as a promising oncolytic platform. Previous studies of MeV entry focused on the identification of cellular receptors. However, the endocytic and trafficking pathways utilized during MeV entry remain poorly described. The contribution of each endocytic pathway has been examined in cells that express the MeV receptors SLAM (signaling lymphocyte-activating molecule) and PVRL4 (poliovirus receptor-like 4) (nectin-4). Recombinant MeVs expressing either firefly luciferase or green fluorescent protein together with a variety of inhibitors were used. The results showed that MeV uptake was dynamin independent in the Vero.hPVRL4, Vero.hSLAM, and PVRL4-positive MCF7 breast cancer cell lines. However, MeV infection was blocked by 5-(N-ethyl-N-propyl)amiloride (EIPA), the hallmark inhibitor of macropinocytosis, as well as inhibitors of actin polymerization. By using phalloidin staining, MeV entry was shown to induce actin rearrangements and the formation of membrane ruffles accompanied by transient elevated fluid uptake. Small interfering RNA (siRNA) knockdown of p21-activated kinase 1 (PAK1) demonstrated that MeV enters both Vero.hPVRL4 and Vero.hSLAM cells in a PAK1-independent manner using a macropinocytosis-like pathway. In contrast, MeV entry into MCF7 human breast cancer cells relied upon Rac1 and its effector PAK1 through a PVRL4-mediated macropinocytosis pathway. MeV entry into DLD-1 colon and HTB-20 breast cancer cells also appeared to use the same pathway. Overall, these findings provide new insight into the life cycle of MeV, which could lead to therapies that block virus entry or methods that improve the uptake of MeV by cancer cells during oncolytic therapy. IMPORTANCE In the past decades, measles virus (MeV) has emerged as a promising oncolytic platform. Previous studies concerning MeV entry focused mainly on the identification of putative receptors for MeV. Nectin-4 (PVRL4) was recently identified as the epithelial cell receptor for MeV. However, the specific endocytic and trafficking pathways utilized during MeV infections are poorly documented. In this study, we demonstrated that MeV enters host cells via a dynamin-independent and actin-dependent endocytic pathway. Moreover, we show that MeV gains entry into MCF7, DLD-1, and HTB-20 cancer cells through a PVRL4-mediated macropinocytosis pathway and identified the typical cellular GTPase and kinase involved. Our findings provide new insight into the life cycle of MeV, which may lead to the development of therapies that block the entry of the virus into the host cell or alternatively promote the uptake of oncolytic MeV into cancer cells. PMID:28250131

Notch Signaling Regulates Late-Stage Epidermal Differentiation and Maintains Postnatal Hair Cycle Homeostasis

PubMed Central

Lin, Hsien-Yi; Kao, Cheng-Heng; Lin, Kurt Ming-Chao; Kaartinen, Vesa; Yang, Liang-Tung

2011-01-01

Background Notch signaling involves ligand-receptor interactions through direct cell-cell contact. Multiple Notch receptors and ligands are expressed in the epidermis and hair follicles during embryonic development and the adult stage. Although Notch signaling plays an important role in regulating differentiation of the epidermis and hair follicles, it remains unclear how Notch signaling participates in late-stage epidermal differentiation and postnatal hair cycle homeostasis. Methodology and Principal Findings We applied Cre/loxP system to generate conditional gene targeted mice that allow inactivation of critical components of Notch signaling pathway in the skin. Rbpj, the core component of all four Notch receptors, and Pofut1, an essential factor for ligand-receptor interactions, were inactivated in hair follicle lineages and suprabasal layer of the epidermis using the Tgfb3-Cre mouse line. Rbpj conditional inactivation resulted in granular parakeratosis and reactive epidermal hyperplasia. Pofut1 conditional inactivation led to ultrastructural abnormalities in the granular layer and altered filaggrin processing in the epidermis, suggesting a perturbation of the granular layer differentiation. Disruption of Pofut1 in hair follicle lineages resulted in aberrant telogen morphology, a decrease of bulge stem cell markers, and a concomitant increase of K14-positive keratinocytes in the isthmus of mutant hair follicles. Pofut1-deficent hair follicles displayed a delay in anagen re-entry and dysregulation of proliferation and apoptosis during the hair cycle transition. Moreover, increased DNA double stand breaks were detected in Pofut1-deficent hair follicles, and real time PCR analyses on bulge keratinocytes isolated by FACS revealed an induction of DNA damage response and a paucity of DNA repair machinery in mutant bulge keratinocytes. Significance our data reveal a role for Notch signaling in regulating late-stage epidermal differentiation. Notch signaling is required for postnatal hair cycle homeostasis by maintaining proper proliferation and differentiation of hair follicle stem cells. PMID:21267458

Does the end justify the means? The contemporary role of dissection/re-entry strategies for recanalization of coronary chronic total occlusions.

PubMed

Karatasakis, Aris; Brilakis, Emmanouil S

2017-11-01

Antegrade and retrograde dissection/re-entry techniques are frequently utilized in contemporary CTO PCI, especially for complex lesions. One-year outcomes with modern dissection/re-entry techniques appear favorable and comparable with those achieved after intraplaque crossing, supporting their increased use. Randomized data on the procedural safety, efficiency, and long-term outcomes of subadventitial CTO PCI techniques are needed. © 2017 Wiley Periodicals, Inc.

New re-entry device for revascularization of chronic coronary total occlusions: preliminary single Japanese center experience.

PubMed

Tsuchikane, Etsuo; Kimura, Masashi; Suzuki, Takahiko; Habara, Maoto; Kurita, Tairo; Tanaka, Nobuyoshi; Nasu, Kenya; Ito, Tatsuya; Kinoshita, Yoshihisa; Wyman, R Michael

2012-08-01

Although retrograde approach for coronary chronic total occlusion (CTO) has been introduced, the procedure is still time and resource consuming. A simplified antegrade approach mightbe another resort. The aim of this study was to evaluate a new device designed to facilitate guidewire re-entry into the true lumen of a CTO from the adjacent subintimal space. Patients with CTO were entered into a prospective registry regardless of lesion characteristics. A new metal-tip catheter was used initially in primary use cases. If it created subintimal tracking, a new re-entry tool (a flat balloon with 2 exit ports offset by 180 degrees) was used as a platform to attempt guidewire penetration into the distal true lumen. In rescue use cases after unsuccessful conventional wiring, the re-entry procedure was subsequently attempted. In 11 CTO lesions attempted, device success was achieved in 8 cases (72.7%). Re-entry procedure success rate was higher in primary use cases (80%) compared to rescue use cases (33.3%). Retrograde approach was conducted immediately after unsuccessful antegrade procedure using this device in the other 3 cases and successful recanalization was achieved in all cases. All lesions were stented, resulting in TIMI 3 flow without major complications. A new coronary re-entry device may provide another strategic option in the antegrade approach to recanalize CTOs.

Ongoing Capabilities and Developments of Re-Entry Plasma Ground Tests at EADS-ASTRIUM

NASA Technical Reports Server (NTRS)

Jullien, Pierre

2008-01-01

During re-entry, spacecrafts are subjected to extreme thermal loads. On mars, they may go through dust storms. These external heat loads are leading the design of re-entry vehicles or are affecting it for spacecraft facing solid propellant jet stream. Sizing the Thermal Protection System require a good knowledge of such solicitations and means to model and reproduce them on earth. Through its work on European projects, ASTRIUM has developed the full range of competences to deal with such issues. For instance, we have designed and tested the heat-shield of the Huygens probe which landed on Titan. In particular, our plasma generators aim to reproduce a wide variety of re-entry conditions. Heat loads are generated by the huge speed of the probes. Such conditions cannot be fully reproduced. Ground tests focus on reproducing local aerothermal loads by using slower but hotter flows. Our inductive plasma torch enables to test little samples at low TRL. Amongst the arc-jets, one was design to test architecture design of ISS crew return system and others fit more severe re-entry such as sample returns or Venus re-entry. The last developments aimed in testing samples in seeded flows. First step was to design and test the seeding device. Special diagnostics characterizing the resulting flow enabled us to fit it to the requirements.

78 FR 40531 - Self-Regulatory Organizations; BATS Exchange, Inc.; Notice of Filing of a Proposed Rule Change...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-07-05

... Exchange from allowing re-entry into the Program where the Exchange deems such re-entry as proper. CLP... its status as a CLP, such Member may re- apply for CLP status. Such application process shall occur at...

A cell stabilization factor for transport of experimental cell cultures to and from the International Space Station

NASA Astrophysics Data System (ADS)

Fattaey, Heideh K.; Consigli, Richard A.; Grenz, Ladonna; Johnson, Terry C.

2000-01-01

The requirement for long term storage of cell cultures previous to arrival on the International Space Station (ISS), as well as culture maintenance after the conduct of experiment in microgravity, necessitates inhibition of cell proliferation and metabolism pending return to earth-based laboratories. Transport of cells in a nonstabilized condition can lead to a loss of cell viability and/or a source of selection pressures for survival that can alter the overall cell population. We have isolated in our laboratory a reversible inhibitor of cell proliferation, a cell regulatory sialoglycopeptide (CeReS-18), that has the capability of stabilizing cells isolated from a wide phylogenetic range by arresting them in the G1 phase of the cell cycle. We show here that CeReS-18 is unusually stable and can be stored at ambient temperatures for weeks without a measurable loss in its biological activity. In addition we demonstrate that CeReS-18 is a superior cell-stabilizing agent as compared to other methods deployed for cell stabilization purposes, such as, decrease in the incubation temperature and serum down shifts. We also discovered that hybridoma cultures stabilized in their proliferative cycle by CeReS-18 produced 150%-300% more antibody per cell than that measured in the proliferating control cultures. The reversible inhibitory activity of CeReS-18, together with its unusual stability, as well as its wide target range lend themselves to use of this inhibitor as a cell stabilizing agent for cell transport to and storage on the ISS. .

Two LXXLL motifs in the N terminus of Mps1 are required for Mps1 nuclear import during G(2)/M transition and sustained spindle checkpoint responses.

PubMed

Zhang, Xiaojuan; Yin, Qingqing; Ling, Youguo; Zhang, Yanhong; Ma, Runlin; Ma, Qingjun; Cao, Cheng; Zhong, Hui; Liu, Xuedong; Xu, Quanbin

2011-08-15

Spindle assembly checkpoint kinase Mps1 is spatially and temporally regulated during cell cycle progression. Mps1 is predominately localized to the cytosol in interphase cells, whereas it is concentrated on kinetochores in prophase and prometaphase cells. The timing and mechanism of Mps1 redistribution during cell cycle transition is currently poorly understood. Here, we show that Mps1 relocates from the cytosol to the nucleus at the G 2/M boundary prior to nuclear envelope breakdown (NEB). This timely translocation depends on two tandem LXXLL motifs in the N terminus of Mps1, and mutations in either motif abolish Mps1 nuclear accumulation. Furthermore, we found that phosphorylation of Mps1 Ser80 (which is located between the two LXXLL motifs) also plays a role in regulating timely nuclear entry of Mps1. Mps1 that is defective in LXXLL motifs has near wild-type kinase activity. Moreover, the kinase activity of Mps1 appears to be dispensable for nuclear translocation, as inhibition of Mps1 by a highly specific small-molecule inhibitor did not perturb its nuclear entry. Remarkably, translocation-deficient Mps1 can mediate activation of spindle assembly checkpoint response; however, it fails to support a sustained mitotic arrest upon prolonged treatment with nocodazole. The mitotic slippage can be attributed to precocious degradation of Mps1 in the arrested cells. Our studies reveal a novel cell cycle-dependent nuclear translocation signal in the N terminus of Mps1 and suggest that timely nuclear entry could be important for sustaining spindle assembly checkpoint responses.

Two LXXLL motifs in the N terminus of Mps1 are required for Mps1 nuclear import during G2/M transition and sustained spindle checkpoint responses

PubMed Central

Zhang, Xiaojuan; Yin, Qingqing; Ling, Youguo; Zhang, Yanhong; Ma, Runlin; Ma, Qingjun; Cao, Cheng; Zhong, Hui

2011-01-01

Spindle assembly checkpoint kinase Mps1 is spatially and temporally regulated during cell cycle progression. Mps1 is predominately localized to the cytosol in interphase cells, whereas it is concentrated on kinetochores in prophase and prometaphase cells. The timing and mechanism of Mps1 redistribution during cell cycle transition is currently poorly understood. Here, we show that Mps1 relocates from the cytosol to the nucleus at the G2/M boundary prior to nuclear envelope breakdown (NEB). This timely translocation depends on two tandem LXXLL motifs in the N terminus of Mps1, and mutations in either motif abolish Mps1 nuclear accumulation. Furthermore, we found that phosphorylation of Mps1 Ser80 (which is located between the two LXXLL motifs) also plays a role in regulating timely nuclear entry of Mps1. Mps1 that is defective in LXXLL motifs has near wild-type kinase activity. Moreover, the kinase activity of Mps1 appears to be dispensable for nuclear translocation, as inhibition of Mps1 by a highly specific small-molecule inhibitor did not perturb its nuclear entry. Remarkably, translocation-deficient Mps1 can mediate activation of spindle assembly checkpoint response; however, it fails to support a sustained mitotic arrest upon prolonged treatment with nocodazole. The mitotic slippage can be attributed to precocious degradation of Mps1 in the arrested cells. Our studies reveal a novel cell cycle-dependent nuclear translocation signal in the N terminus of Mps1 and suggest that timely nuclear entry could be important for sustaining spindle assembly checkpoint responses. PMID:21778823

Live-cell imaging visualizes frequent mitotic skipping during senescence-like growth arrest in mammary carcinoma cells exposed to ionizing radiation.

PubMed

Suzuki, Masatoshi; Yamauchi, Motohiro; Oka, Yasuyoshi; Suzuki, Keiji; Yamashita, Shunichi

2012-06-01

Senescence-like growth arrest in human solid carcinomas is now recognized as the major outcome of radiotherapy. This study was designed to analyze cell cycle during the process of senescence-like growth arrest in mammary carcinoma cells exposed to X-rays. Fluorescent ubiquitination-based cell cycle indicators were introduced into the human mammary carcinoma cell line MCF-7. Cell cycle was sequentially monitored by live-cell imaging for up to 5 days after exposure to 10 Gy of X-rays. Live-cell imaging revealed that cell cycle transition from G2 to G1 phase without mitosis, so-called mitotic skipping, was observed in 17.1% and 69.8% of G1- and G2-irradiated cells, respectively. Entry to G1 phase was confirmed by the nuclear accumulation of mKO(2)-hCdt1 as well as cyclin E, which was inversely correlated to the accumulation of G2-specific markers such as mAG-hGeminin and CENP-F. More than 90% of cells skipping mitosis were persistently arrested in G1 phase and showed positive staining for the senescent biochemical marker, which is senescence-associated ß-galactosidase, indicating induction of senescence-like growth arrest accompanied by mitotic skipping. While G2 irradiation with higher doses of X-rays induced mitotic skipping in approximately 80% of cells, transduction of short hairpin RNA (shRNA) for p53 significantly suppressed mitotic skipping, suggesting that ionizing radiation-induced mitotic skipping is associated with p53 function. The present study found the pathway of senescence-like growth arrest in G1 phase without mitotic entry following G2-irradiation. Copyright © 2012 Elsevier Inc. All rights reserved.

8 CFR 212.1 - Documentary requirements for nonimmigrants.

Code of Federal Regulations, 2011 CFR

2011-01-01

... Cards and a valid Taiwan passport with a valid re-entry permit issued by the Taiwan Ministry of Foreign... valid re-entry permit issued by the Taiwan Ministry of Foreign Affairs. (2) Program Countries and... the United States from contiguous territory or adjacent islands at a land or sea port-of-entry. A...

8 CFR 212.1 - Documentary requirements for nonimmigrants.

Code of Federal Regulations, 2014 CFR

2014-01-01

... Cards and a valid Taiwan passport with a valid re-entry permit issued by the Taiwan Ministry of Foreign... Taiwan National Identity Card and a valid Taiwan passport with a valid re-entry permit issued by the... the United States from contiguous territory or adjacent islands at a land or sea port-of-entry. A...

8 CFR 212.1 - Documentary requirements for nonimmigrants.

Code of Federal Regulations, 2012 CFR

2012-01-01

... Cards and a valid Taiwan passport with a valid re-entry permit issued by the Taiwan Ministry of Foreign... Taiwan National Identity Card and a valid Taiwan passport with a valid re-entry permit issued by the... the United States from contiguous territory or adjacent islands at a land or sea port-of-entry. A...

8 CFR 212.1 - Documentary requirements for nonimmigrants.

Code of Federal Regulations, 2013 CFR

2013-01-01

... Cards and a valid Taiwan passport with a valid re-entry permit issued by the Taiwan Ministry of Foreign... Taiwan National Identity Card and a valid Taiwan passport with a valid re-entry permit issued by the... the United States from contiguous territory or adjacent islands at a land or sea port-of-entry. A...

Microgravity can activate signals urging cells to S-phase entry during tissue and organ regeneration in Urodele amphibians exposed to real and simulated microgravity

NASA Astrophysics Data System (ADS)

Grigoryan, E.; Anton, H.-J.; Mitashov, V.

Regenerative response following local injury or tissue removal in urodele amphibians is dependent on cell cycle entry of cells sources for regeneration in the remaining tissue. In a number of our experiments performed aboard biosatellites in orbital flights and fast rotated clinostat we found enhanced proliferative activity and, as a result, regeneration quicker than that in controls. In each investigated case an activity of cell proliferation evaluated by 3H-thymidine radioautography and BrdU assay at the early stages of lens, retina, forelimb and tail regeneration in newts was about 1,2-1,7 fold higher both under conditions of real and physiological weightlessness as compared with controls. Faster S-phase entry under conditions of micro- g was demonstrated by cycling multipotent cells as well as by differentiated postmitotic cells both participated in regeneration. Important, that cycling cells outside areas of regeneration were also found as displayed faster cellular growth. In our papers (1,2,3,4) we offered some hypothesis that could explain mechanisms of low g stimulating effect upon cell growth in regeneration in Urodela. In particular, changes in expression of some growth factors and their receptors, as well as the synthesis of specific range of generalized stress proteins (AGSPs) were proposed. However, in fact, molecular mechanisms of micro- g effect upon cell proliferation are mediated by changes on organismic level induced by micro- g environment. Some of them which are able to trigger off signaling changes on the cellular level that, in turn, evoke cells to grow faster would be represented in our report. 1. Mitashov V. et al. Adv. Space Res. 1996. 17 (6/7): 241-255 2. Anton H.-J. et al. Adv. Space Res. 1996. 17 (6/7): 55-65 3. Grigoryan E. et al. Adv. Space Res. 1998. 22 (2): 293-301 4. Grigoryan E. et al. Adv. Space Res. 2002. 30 (4): 757-764

Division of labour between Myc and G1 cyclins in cell cycle commitment and pace control.

PubMed

Dong, Peng; Maddali, Manoj V; Srimani, Jaydeep K; Thélot, François; Nevins, Joseph R; Mathey-Prevot, Bernard; You, Lingchong

2014-09-01

A body of evidence has shown that the control of E2F transcription factor activity is critical for determining cell cycle entry and cell proliferation. However, an understanding of the precise determinants of this control, including the role of other cell-cycle regulatory activities, has not been clearly defined. Here, recognizing that the contributions of individual regulatory components could be masked by heterogeneity in populations of cells, we model the potential roles of individual components together with the use of an integrated system to follow E2F dynamics at the single-cell level and in real time. These analyses reveal that crossing a threshold amplitude of E2F accumulation determines cell cycle commitment. Importantly, we find that Myc is critical in modulating the amplitude, whereas cyclin D/E activities have little effect on amplitude but do contribute to the modulation of duration of E2F activation, thereby affecting the pace of cell cycle progression.

Utilizing Weather RADAR for Rapid Location of Meteorite Falls and Space Debris Re-Entry

NASA Technical Reports Server (NTRS)

Fries, Marc D.

2016-01-01

This activity utilizes existing NOAA weather RADAR imagery to locate meteorite falls and space debris falls. The near-real-time availability and spatial accuracy of these data allow rapid recovery of material from both meteorite falls and space debris re-entry events. To date, at least 22 meteorite fall recoveries have benefitted from RADAR detection and fall modeling, and multiple debris re-entry events over the United States have been observed in unprecedented detail.

10.2 Thermal-Structural Testing

NASA Technical Reports Server (NTRS)

Hudson, Larry D.

2008-01-01

Objective: Test a C/SiC Ruddervator Subcomponent under relevant thermal, mechanical & dynamic loading a) Thermal-structural mission cycling for re-entry and hypersonic cruise conditions; b) High-temperature modal survey to study the effect of heating on mode shapes, natural frequencies and damping. Supports NASA ARMD Hypersonics Material & Structures Program. Partners: NASA Dryden / Langley / Glenn, Lockheed-Martin, Materials Research & Design, GE CCP Test Phases - Phase 1: Acoustic-Vibration Testing (LaRC) completed - Phase 2: Thermal-Mechanical Testing (DFRC) in assembly - Phase 3: Mechanical Testing (DFRC) in assembly

Federal policy on criminal offenders who have substance use disorders: how can we maximize public health and public safety?

PubMed

Humphreys, Keith

2012-01-01

The Obama Administration is striving to promote both public health and public safety by improving the public policy response to criminal offenders who have substance use disorders. This includes supporting drug courts, evidence-based probation and parole programs, addiction treatment and re-entry programs. Scientists and clinicians in the addiction field have a critical role to play in this much-needed effort to break the cycle of addiction, crime and incarceration.

mei-41 and bub1 block mitosis at two distinct steps in response to incomplete DNA replication in Drosophila embryos.

PubMed

Garner, M; van Kreeveld, S; Su, T T

2001-10-16

Drosophila double park encodes a homolog of Cdt1 that functions in initiation of DNA replication in fission yeast and Xenopus. dup mutants complete the first 15 embryonic cell cycles, presumably via maternal dup products, and show defects in the 16(th) S phase (S16). Cells carrying dup(a1) allele forgo S16 altogether but enter mitosis 16 (M16). We find that the timing of entry into M16 is similar in dup(a1) and heterozygous or wild-type (wt) controls. In contrast, we find that mutant cells carrying another allele, dup(a3), undergo a partial S16 and delay the entry into M16. Thus, initiation of S16 appears necessary for delaying M16. This delay is absent in double mutants of dup(a3) and mei-41 (Drosophila ATR), indicating that a mei-41-dependent checkpoint acts to delay the entry into mitosis in response to incomplete DNA replication. dup(a3) and dup(a1) mutant cells that enter M16 become arrested in M16. We find that mitotic cyclins are stabilized and that a spindle checkpoint protein, Bub1, localizes onto chromosomes during mitotic arrest in dup mutants. These features suggest an arrest prior to metaphase-anaphase transition. dup(a3) bub1 double mutant cells exit M16, indicating that a bub1-mediated checkpoint acts to block mitotic exit in dup mutants. To our knowledge, this is the first report of (1) incomplete DNA replication affecting both the entry into and the exit from mitosis in a single cell cycle via different mechanisms and (2) the role of bub1 in regulating mitotic exit in response to incomplete DNA replication.

Virtual Reality Modelling Simulation of the Re-entry Motion of an Axialsymmetric Vehicle

NASA Astrophysics Data System (ADS)

Guidi, A.; Chu, Q.. P.; Mulder, J. A.

This work started during the stability analysis of the Delft Aerospace Re-entry Test demonstrator (DART) which is a small axisymmetric ballistic re-entry vehicle. The dynamic stability evaluation of an axisymmetric re-entry vehicle is especially concerned on the behaviour of its angle of attack during the flight through the atmosphere. The variation in the angle of attack is essential for prediction of the trajectory of the vehicle and for heating requirement of the structure of the vehicle. The concept of the total angle of attack and the windward meridian plane are introduced. The position of the centre of pressure can be a crucial point in the stability of the vehicle. Although the simpleness of an axisymmetric shape, the re-entry of such a vehicle is characterised by several complex phenomenologies that were analysed with the aid of the flight simulator and of a 3D virtual reality modeling simulator. Simulations were performed with a 25° AOA initial condition in order to simulate the response of the vehicle to a disturbance that may occur during the flight causing a variation in attitude from its Trim . Certain aspects of re-entry vehicle motion are conveniently described in the terms of Euler angles. Using the Eulerian angle it is possible to generate a tridimensional animation of the output of the Flight Simulator. This tridimensional analysis is of great importance in order to understand the mentioned complex motions. Furthermore with growing in computer power it is possible to generate online visualisation of the simulations. The output of the flight simulator was used in a software written in Virtual Reality Modelling Language (VRML). With VRML this software was possible the visualisation of the re-entry motion of the vehicle. With this option the animation can run on-line during the with the flight simulator and can be also easily published on the internet or send to other users in very small file size. (the VRLM simulation of the re-entry, can be seen at the official DART internet site: www.dart-project.com)

EZH2-mediated H3K27 trimethylation mediates neurodegeneration in ataxia-telangiectasia

PubMed Central

Li, Jiali; Hart, Ronald P.; Mallimo, Elyse M.; Swerdel, Mavis R.; Kusnecov, Alexander; Herrup, Karl

2014-01-01

The symptoms of ataxia-telangiectasia (A-T) include a progressive neurodegeneration caused by ATM protein deficiency. We previously found that nuclear accumulation of histone deacetylase-4, HDAC4, contributes to this degeneration; we now report that increased histone H3K27 trimethylation (H3K27me3) mediated by polycomb repressive complex 2 (PRC2) also plays an important role in the A-T phenotype. Enhancer of zeste homolog 2 (EZH2), a core catalytic component of PRC2, is a new ATM kinase target, and ATM-mediated S734 phosphorylation of EZH2 reduces protein stability. Thus, PRC2 formation is elevated along with H3K27me3in ATM deficiency. ChIP-sequencing shows a significant increase in H3K27me3 ‘marks’ and a dramatic shift in their location. The change of H3K27me3 chromatin-binding pattern is directly related to cell cycle re-entry and cell death of ATM-deficient neurons. Lentiviral knockdown of EZH2 rescues Purkinje cell degeneration and behavioral abnormalities in Atm−/− mice, demonstrating that EZH2 hyperactivity is another key factor in A-T neurodegeneration. PMID:24162653

The Temporal Regulation of S Phase Proteins During G1

PubMed Central

Grant, Gavin D.; Cook, Jeanette G.

2018-01-01

Successful DNA replication requires intimate coordination with cell cycle progression. Prior to DNA replication initiation in S phase, a series of essential preparatory events in G1 phase ensures timely, complete, and precise genome duplication. Among the essential molecular processes are regulated transcriptional upregulation of genes that encode replication proteins, appropriate post-transcriptional control of replication factor abundance and activity, and the assembly of DNA-loaded protein complexes to license replication origins. In this chapter we describe these critical G1 events necessary for DNA replication and their regulation in the context of both cell cycle entry and cell cycle progression. PMID:29357066

Entry inhibitors: New advances in HCV treatment

PubMed Central

Qian, Xi-Jing; Zhu, Yong-Zhe; Zhao, Ping; Qi, Zhong-Tian

2016-01-01

Hepatitis C virus (HCV) infection affects approximately 3% of the world's population and causes chronic liver diseases, including liver fibrosis, cirrhosis, and hepatocellular carcinoma. Although current antiviral therapy comprising direct-acting antivirals (DAAs) can achieve a quite satisfying sustained virological response (SVR) rate, it is still limited by viral resistance, long treatment duration, combined adverse reactions, and high costs. Moreover, the currently marketed antivirals fail to prevent graft reinfections in HCV patients who receive liver transplantations, probably due to the cell-to-cell transmission of the virus, which is also one of the main reasons behind treatment failure. HCV entry is a highly orchestrated process involving initial attachment and binding, post-binding interactions with host cell factors, internalization, and fusion between the virion and the host cell membrane. Together, these processes provide multiple novel and promising targets for antiviral therapy. Most entry inhibitors target host cell components with high genetic barriers and eliminate viral infection from the very beginning of the viral life cycle. In future, the addition of entry inhibitors to a combination of treatment regimens might optimize and widen the prevention and treatment of HCV infection. This review summarizes the molecular mechanisms and prospects of the current preclinical and clinical development of antiviral agents targeting HCV entry. PMID:26733381

DOE Office of Scientific and Technical Information (OSTI.GOV)

Suzuki, Masatoshi, E-mail: msuzuki@nagasaki-u.ac.jp; Yamauchi, Motohiro; Oka, Yasuyoshi

Purpose: Senescence-like growth arrest in human solid carcinomas is now recognized as the major outcome of radiotherapy. This study was designed to analyze cell cycle during the process of senescence-like growth arrest in mammary carcinoma cells exposed to X-rays. Methods and Materials: Fluorescent ubiquitination-based cell cycle indicators were introduced into the human mammary carcinoma cell line MCF-7. Cell cycle was sequentially monitored by live-cell imaging for up to 5 days after exposure to 10 Gy of X-rays. Results: Live-cell imaging revealed that cell cycle transition from G2 to G1 phase without mitosis, so-called mitotic skipping, was observed in 17.1% andmore » 69.8% of G1- and G2-irradiated cells, respectively. Entry to G1 phase was confirmed by the nuclear accumulation of mKO{sub 2}-hCdt1 as well as cyclin E, which was inversely correlated to the accumulation of G2-specific markers such as mAG-hGeminin and CENP-F. More than 90% of cells skipping mitosis were persistently arrested in G1 phase and showed positive staining for the senescent biochemical marker, which is senescence-associated ss-galactosidase, indicating induction of senescence-like growth arrest accompanied by mitotic skipping. While G2 irradiation with higher doses of X-rays induced mitotic skipping in approximately 80% of cells, transduction of short hairpin RNA (shRNA) for p53 significantly suppressed mitotic skipping, suggesting that ionizing radiation-induced mitotic skipping is associated with p53 function. Conclusions: The present study found the pathway of senescence-like growth arrest in G1 phase without mitotic entry following G2-irradiation.« less

Art Concept - Apollo VIII - Command Module (CM) - Re-Entry Orientation

NASA Image and Video Library

1968-01-01

S68-55292 (August 1968) --- A North American Rockwell Corporation artist's concept depicting the Apollo Command Module (CM), oriented in a blunt-end-forward attitude, re-entering Earth's atmosphere after returning from a lunar landing mission. Note the change in color caused by the extremely high temperatures encountered upon re-entry.

The circadian molecular clock regulates adult hippocampal neurogenesis by controlling the timing of cell-cycle entry and exit.

PubMed

Bouchard-Cannon, Pascale; Mendoza-Viveros, Lucia; Yuen, Andrew; Kærn, Mads; Cheng, Hai-Ying M

2013-11-27

The subgranular zone (SGZ) of the adult hippocampus contains a pool of quiescent neural progenitor cells (QNPs) that are capable of entering the cell cycle and producing newborn neurons. The mechanisms that control the timing and extent of adult neurogenesis are not well understood. Here, we show that QNPs of the adult SGZ express molecular-clock components and proliferate in a rhythmic fashion. The clock proteins PERIOD2 and BMAL1 are critical for proper control of neurogenesis. The absence of PERIOD2 abolishes the gating of cell-cycle entrance of QNPs, whereas genetic ablation of bmal1 results in constitutively high levels of proliferation and delayed cell-cycle exit. We use mathematical model simulations to show that these observations may arise from clock-driven expression of a cell-cycle inhibitor that targets the cyclin D/Cdk4-6 complex. Our findings may have broad implications for the circadian clock in timing cell-cycle events of other stem cell populations throughout the body. Copyright © 2013 The Authors. Published by Elsevier Inc. All rights reserved.

Successful revascularization of chronic total occlusion of lower extremity arteries: a wire only and bail out use of re-entry device approach.

PubMed

Langhoff, R; Stumpe, S; Treitl, M; Schulte, K L

2013-10-01

The management of progressive peripheral artery disease experienced a vast change in paradigms over the last decades for the benefit of minimal invasive therapy as a first-line strategy. With the constant development of new devices, materials and dedicated access strategies, more complex lesions can be managed but the limitations to successfully treat chronic total occlusions are still the challenge to re-enter the true lumen. The aim of this retrospective study was to investigate, if a "wire only" strategy leads to an acceptable success rate in a mixed cohort of CTO lesions and to what extend re-entry devices are used. We retrospectively analyzed patients treated at the Vascular Center Berlin between 2011 and 2013 with chronic total occlusion out of a prospective conducted database (Endovascular MILestones - EMIL) for demographics, risk factors, co-morbidities, technical success rates, lesion characteristics and use of guidewires as well as re-entry systems. A total of 128 patients with 146 lesions, which represent a subgroup of all the cases performed in our center, following a predefined treatment algorithm for chronic total occlusions (CTOs), have been analyzed. We achieved a technical success in 133 (91.1%) of all cases following a "wire only" strategy. Out of 13 (8.9%) CTOs with technical failure in 7 (53.9%) CTOs a re-entry device (Off-Road®) with a 100% technical success has been used. In 91.1% of chronic total occlusion lesions the use of 2 wires only (88.7%) led to a successful recanalization. A "wire only" strategy followed by the use of a re-entry device as a bail out strategy, led to a total of 140 (96%) lesions to be successfully recanalized. In more than 90% of all cases with chronic total occlusion of peripheral lower extremity arteries, endovascular intervention has been successful following a "wire only" strategy. When deciding to use a re-entry device, in case of a failure of a proper wire re-entry at the reconstitution point, a technical success rate of 100% was achieved. Therefore following a strict wire algorithm and considering the use of a re-entry system as a bail out strategy will lead to a successful minimal invasive management of chronic total occlusion in nearly 100% of the cases with TASC II A - D lesions.

Incidence and outcome of re-entry injury in redo cardiac surgery: benefits of preoperative planning.

PubMed

Imran Hamid, Umar; Digney, Ruairi; Soo, Lorraine; Leung, Samantha; Graham, Alastair N J

2015-05-01

Repeat sternotomy for redo cardiac surgery may be associated with catastrophic injuries to mediastinal structures. The purpose of this study was to determine the frequency of these injuries, associated outcome and if a preoperative computerized tomography (CT) scan reduces the risk of re-entry injury. Five hundred and forty-four patients who underwent redo cardiac surgery between 2001 and 2011 were identified by review of our unit's prospectively maintained cardiac surgery database. Demographic details, surgical strategy, re-entry injuries, hospital stay, in-hospital mortality and long-term survival were analysed. The mean age was 61 years; 326 were male, 218 were female. Four hundred and eighty six patients underwent first time redo surgery, while 58 patients had multiple previous operations. The median logistic EuroSCORE was 11, in-hospital mortality rate was 9.5% and observed to expected mortality rate was 0.8. Re-entry complications occurred in 15 cases (2.7%). These included injuries to the aorta (n = 2), right atrium (n = 1), innominate vein (n = 2), internal mammary artery (n = 2), pulmonary artery (n = 2), lung parenchyma (n = 1), saphenous vein graft (n = 2), right ventricle (n = 2) and ventricular fibrillation (n = 1). The mortality rate in patients with re-entry injury was 26% (n = 4) compared with 9% (n = 48) in those without re-entry complications. Preoperative planning by CT scan was performed in 162 cases and adherence of vital structures to the sternum was found in 60 cases; the right ventricle, innominate vein and bypass grafts in 41, 11 and 8, respectively. The incidence rate of re-entry injury was 0.6% in these patients vs 3.6% in those who did not have a preoperative CT scan (P = 0.046). Peripheral arterial cannulation was carried out in 35 patients (6.4%) to establish cardiopulmonary bypass (CPB) prior to sternotomy, and there were no mediastinal injuries observed in these cases. Multivariate logistic regression analysis revealed re-entry injury as one of the independent predictors of in-hospital mortality (P = 0.039). The incidence of re-entry injury during repeat sternotomy is low; however, it is associated with a significant increase in the risk of in-hospital mortality. Preoperative planning using CT scan reduces the risk by identifying adherent structures, and, in selected patients, establishing CPB prior to sternotomy is a safe strategy in redo cardiac surgery. © The Author 2014. Published by Oxford University Press on behalf of the European Association for Cardio-Thoracic Surgery. All rights reserved.

Prevalence and correlates of community re-entry challenges faced by HIV-infected male prisoners in Malaysia

PubMed Central

Choi, P; Kavasery, R; Desai, M M; Govindasamy, S; Kamarulzaman, A; Altice, F L

2010-01-01

Summary HIV-infected prisoners face an inordinate number of community re-entry challenges. In 2007, 102 HIV-infected prisoners in Malaysia were surveyed anonymously within six months prior to release to assess the prevalence and correlates of community re-entry challenges. Staying out of prison (60.8%), remaining off drugs (39.2%), finding employment (35.3%) and obtaining HIV care (32.4%) were the re-entry challenges reported most frequently. Global stigma, negative self-image and public attitudes-related stigma were independently associated with challenges to obtaining HIV care. In multivariate analyses, those with previous incarcerations (adjusted odds ratio [AOR], 3.2; 95% confidence interval [CI], 1.4–7.6), higher HIV-related symptoms (AOR, 2.0; 95% CI, 1.0–4.1) and higher public attitudes-related stigma (AOR, 2.5; 95% CI, 1.2–5.1) had a significantly higher likelihood of identifying more re-entry challenges. Targeted interventions, such as effective drug treatment, HIV care and public awareness campaigns, are crucial for stemming the HIV epidemic and improving health outcomes among HIV-infected prisoners in Malaysia. PMID:20606222

GRP78 Is an Important Host Factor for Japanese Encephalitis Virus Entry and Replication in Mammalian Cells.

PubMed

Nain, Minu; Mukherjee, Sriparna; Karmakar, Sonali Porey; Paton, Adrienne W; Paton, James C; Abdin, M Z; Basu, Anirban; Kalia, Manjula; Vrati, Sudhanshu

2017-03-15

Japanese encephalitis virus (JEV), a mosquito-borne flavivirus, is the leading cause of viral encephalitis in Southeast Asia with potential to become a global pathogen. Here, we identify glucose-regulated protein 78 (GRP78) as an important host protein for virus entry and replication. Using the plasma membrane fractions from mouse neuronal (Neuro2a) cells, mass spectroscopy analysis identified GRP78 as a protein interacting with recombinant JEV envelope protein domain III. GRP78 was found to be expressed on the plasma membranes of Neuro2a cells, mouse primary neurons, and human epithelial Huh-7 cells. Antibodies against GRP78 significantly inhibited JEV entry in all three cell types, suggesting an important role of the protein in virus entry. Depletion of GRP78 by small interfering RNA (siRNA) significantly blocked JEV entry into Neuro2a cells, further supporting its role in virus uptake. Immunofluorescence studies showed extensive colocalization of GRP78 with JEV envelope protein in virus-infected cells. This interaction was also confirmed by immunoprecipitation studies. Additionally, GRP78 was shown to have an important role in JEV replication, as treatment of cells post-virus entry with subtilase cytotoxin that specifically cleaved GRP78 led to a substantial reduction in viral RNA replication and protein synthesis, resulting in significantly reduced extracellular virus titers. Our results indicate that GRP78, an endoplasmic reticulum chaperon of the HSP70 family, is a novel host factor involved at multiple steps of the JEV life cycle and could be a potential therapeutic target. IMPORTANCE Recent years have seen a rapid spread of mosquito-borne diseases caused by flaviviruses. The flavivirus family includes West Nile, dengue, Japanese encephalitis, and Zika viruses, which are major threats to public health with potential to become global pathogens. JEV is the major cause of viral encephalitis in several parts of Southeast Asia, affecting a predominantly pediatric population with a high mortality rate. This study is focused on identification of crucial host factors that could be targeted to cripple virus infection and ultimately lead to development of effective antivirals. We have identified a cellular protein, GRP78, that plays a dual role in virus entry and virus replication, two crucial steps of the virus life cycle, and thus is a novel host factor that could be a potential therapeutic target. Copyright © 2017 American Society for Microbiology.

ANNOTATED BIBLIOGRAPHY OF ASTRODYNAMICS AND RE-ENTRY MECHANICS,

DTIC Science & Technology

A selected list of references in the fields of astronautics and re-entry mechanics is classified and discussed, and a comprehensive subject and author index is included for ease in locating the references. (Author)

Involvement of a Gardos-type potassium channel in head activator-induced mitosis of BON cells.

PubMed

Kayser, S T; Ulrich, H; Schaller, H C

1998-06-01

The human neuroendocrine cell line BON was used to study second messengers involved in signal transduction for entry into mitosis. BON cells produce the neuropeptide head activator (HA) and use it as autocrine growth factor. HA stimulates BON cell proliferation by triggering entry into mitosis. HA-induced mitosis is mediated by an inhibitory G protein, the action of which is blocked by pertussis toxin. HA signaling requires inhibition of the cAMP pathway, calcium influx, and hyperpolarization of cells. The latter is a very important and sensitive step involving a calcium-activated potassium channel. Cell cycle progression and proliferation of BON cells are most efficiently inhibited with specific inhibitors of this potassium channel. Pharmacology and RNA analysis suggest identity with the recently cloned Gardos-type potassium channel.

The Secret of Guided Missile Re-Entry,

DTIC Science & Technology

1986-06-25

I RD-PAI169 598 THE SECRET OF GUIDED MISSILE RE-ENTRY(U) FOREIGN / I TECHNOLOGY DIV NRIGHT-PATTERSON RFB OH J CHEN ET AL. I 25 JUN 96 FTD-ID(RS)T...TECHNOLOGY DIVISION THE SECRET OF GUIDED MISSILE RE-ENTRY by Chen Jingzhong, An Sehua J L 0 7 ’:;85’ ’ 0 *Approved for public release; Distribution...unlimite t d. :. 86 7 034.. FTD- ID(RS)T-0459-86 HUMAN TRANSLATION FTD-ID(RS)T-0459-86 25 June 1986 MICROFICHE NR: F - - 0Q 9? THE SECRET OF GUIDED

Missile Aerodynamics for Ascent and Re-entry

NASA Technical Reports Server (NTRS)

Watts, Gaines L.; McCarter, James W.

2012-01-01

Aerodynamic force and moment equations are developed for 6-DOF missile simulations of both the ascent phase of flight and a tumbling re-entry. The missile coordinate frame (M frame) and a frame parallel to the M frame were used for formulating the aerodynamic equations. The missile configuration chosen as an example is a cylinder with fixed fins and a nose cone. The equations include both the static aerodynamic coefficients and the aerodynamic damping derivatives. The inclusion of aerodynamic damping is essential for simulating a tumbling re-entry. Appended information provides insight into aerodynamic damping.

In Their Own Voices: Breaking the Vicious Cycle of Addiction, Treatment and Criminal Justice Among People who Inject Drugs in Ukraine.

PubMed

Mazhnaya, Alyona; Bojko, Martha J; Marcus, Ruthanne; Filippovych, Sergii; Islam, Zahedsul; Dvoriak, Sergey; Altice, Frederick L

To understand how perceived law enforcement policies and practices contribute to the low rates of utilization of opioid agonist therapies (OAT) among people who inject drugs (PWIDs) in Ukraine. Qualitative data from 25 focus groups (FGs) with 199 opioid-dependent PWIDs in Ukraine examined domains related to lived or learned experiences with OAT, police, arrest, incarceration, and criminal activity were analyzed using grounded theory principles. Most participants were male (66%), in their late 30s, and previously incarcerated (85%) mainly for drug-related activities. When imprisoned, PWIDs perceived themselves as being "addiction-free". After prison-release, the confluence of police surveillance, societal stress contributed to participants' drug use relapse, perpetuating a cycle of searching for money and drugs, followed by re-arrest and re-incarceration. Fear of police and arrest both facilitated OAT entry and simultaneously contributed to avoiding OAT since system-level requirements identified OAT clients as targets for police harassment. OAT represents an evidence-based option to 'break the cycle', however, law enforcement practices still thwart OAT capacity to improve individual and public health. In the absence of structural changes in law enforcement policies and practices in Ukraine, PWIDs will continue to avoid OAT and perpetuate the addiction cycle with high imprisonment rates.

Hydrodynamic boost: a novel re-entry technique in subintimal angioplasty of below-the-knee vessels.

PubMed

Ferraresi, Roberto; Hamade, Meneme; Gallicchio, Vito; Troisi, Nicola; Mauri, Giovanni

2016-08-01

To describe the hydrodynamic boost (HB) technique and report our preliminary results with this technique in the subintimal angioplasty of below-the-knee vessels. HB was used in 23 cases (14 males, mean age 73 ± 12 years) of critical limb ischemia, with long chronic total occlusion of tibial arteries extended to the ankle level. The operator performs a manual injection of diluted contrast dye through a 4 F catheter into the subintimal space, close to the patent true distal lumen, in order to achieve a tear in the intimal flap and a connection with the true lumen. In 19/23 (83 %) cases, the HB was effective in creating a connection between the subintimal space and the true distal lumen and it was possible to advance a wire and to conclude the procedure. In 4/23 (17 %) lesions, the HB failed and the procedure was successfully completed by retrograde approach. No major complications occurred. Mean length between catheter tip and re-entry point was 8 ± 5 mm. HB seems to be a feasible, safe and effective re-entry technique in distal below-the-knee vessels. This method represents an easy option for re-entry that extends the possibility of antegrade approach to obtain a successful revascularization. • In subintimal angioplasty of below-the-knee vessel re-entry can represent a challenge. • Inability to re-enter may determine the failure of the revascularization procedure. • HB is a novel re-entry technique feasible in distal below-the-knee vessels. • HB may increase the success rate of antegrade approach. • In case of failure, retrograde approach remains feasible.

DNA hypomethylation induces a DNA replication-associated cell cycle arrest to block hepatic outgrowth in uhrf1 mutant zebrafish embryos

PubMed Central

Jacob, Vinitha; Chernyavskaya, Yelena; Chen, Xintong; Tan, Poh Seng; Kent, Brandon; Hoshida, Yujin; Sadler, Kirsten C.

2015-01-01

UHRF1 (ubiquitin-like, containing PHD and RING finger domains, 1) recruits DNMT1 to hemimethylated DNA during replication and is essential for maintaining DNA methylation. uhrf1 mutant zebrafish have global DNA hypomethylation and display embryonic defects, including a small liver, and they die as larvae. We make the surprising finding that, despite their reduced organ size, uhrf1 mutants express high levels of genes controlling S-phase and have many more cells undergoing DNA replication, as measured by BrdU incorporation. In contrast to wild-type hepatocytes, which are continually dividing during hepatic outgrowth and thus dilute the BrdU label, uhrf1 mutant hepatocytes retain BrdU throughout outgrowth, reflecting cell cycle arrest. Pulse-chase-pulse experiments with BrdU and EdU, and DNA content analysis indicate that uhrf1 mutant cells undergo DNA re-replication and that apoptosis is the fate of many of the re-replicating and arrested hepatocytes. Importantly, the DNA re-replication phenotype and hepatic outgrowth failure are preceded by global loss of DNA methylation. Moreover, uhrf1 mutants are phenocopied by mutation of dnmt1, and Dnmt1 knockdown in uhrf1 mutants enhances their small liver phenotype. Together, these data indicate that unscheduled DNA replication and failed cell cycle progression leading to apoptosis are the mechanisms by which DNA hypomethylation prevents organ expansion in uhrf1 mutants. We propose that cell cycle arrest leading to apoptosis is a strategy that restricts propagation of epigenetically damaged cells during embryogenesis. PMID:25564650

DNA hypomethylation induces a DNA replication-associated cell cycle arrest to block hepatic outgrowth in uhrf1 mutant zebrafish embryos.

PubMed

Jacob, Vinitha; Chernyavskaya, Yelena; Chen, Xintong; Tan, Poh Seng; Kent, Brandon; Hoshida, Yujin; Sadler, Kirsten C

2015-02-01

UHRF1 (ubiquitin-like, containing PHD and RING finger domains, 1) recruits DNMT1 to hemimethylated DNA during replication and is essential for maintaining DNA methylation. uhrf1 mutant zebrafish have global DNA hypomethylation and display embryonic defects, including a small liver, and they die as larvae. We make the surprising finding that, despite their reduced organ size, uhrf1 mutants express high levels of genes controlling S-phase and have many more cells undergoing DNA replication, as measured by BrdU incorporation. In contrast to wild-type hepatocytes, which are continually dividing during hepatic outgrowth and thus dilute the BrdU label, uhrf1 mutant hepatocytes retain BrdU throughout outgrowth, reflecting cell cycle arrest. Pulse-chase-pulse experiments with BrdU and EdU, and DNA content analysis indicate that uhrf1 mutant cells undergo DNA re-replication and that apoptosis is the fate of many of the re-replicating and arrested hepatocytes. Importantly, the DNA re-replication phenotype and hepatic outgrowth failure are preceded by global loss of DNA methylation. Moreover, uhrf1 mutants are phenocopied by mutation of dnmt1, and Dnmt1 knockdown in uhrf1 mutants enhances their small liver phenotype. Together, these data indicate that unscheduled DNA replication and failed cell cycle progression leading to apoptosis are the mechanisms by which DNA hypomethylation prevents organ expansion in uhrf1 mutants. We propose that cell cycle arrest leading to apoptosis is a strategy that restricts propagation of epigenetically damaged cells during embryogenesis. © 2015. Published by The Company of Biologists Ltd.

Obtaining A Degree: Alternative Options for Re-Entry Women. Field Evaluation Draft.

ERIC Educational Resources Information Center

Fisher-Thompson, Jeanne

Problems and barriers that women often face when re-entering the higher educational system are identified, and ways in which institutions can be more responsive to re-entry students are suggested. A wide range of possible actions is included so that institutions can pursue those most appropriate to their individual circumstances. Definitions are…

Intravascular ultrasound guided wiring re-entry technique for complex chronic total occlusions.

PubMed

Huang, Wei-Chieh; Teng, Hsin-I; Hsueh, Chien-Hung; Lin, Shing-Jong; Chan, Wan-Leong; Lu, Tse-Min

2018-05-03

The successful recanalization rate of chronic total occlusion (CTO) lesions without retrograde collaterals available is always low. Intravascular ultrasound (IVUS) may be useful to guide the subintimal guidewire to re-enter the true lumen. We evaluated the clinical feasibility and efficacy of the IVUS-guided wiring re-entry technique for these complex CTO lesions. Twenty consecutive patients (19 male, mean age: 65.3 ± 12.8 years) with both failed antegrade and retrograde approaches were enrolled. The IVUS catheter was introduced into the subintimal space to identify the entry point into the subintimal space, and guide another stiff wire to re-enter the true lumen with the adjacent side-branch or first wire as markers, or using IVUS-guided parallel wire technique. The entry point into the subintimal space was identified by IVUS in all cases, and the IVUS-guided wiring re-entry technique succeeded in 17 cases (85%). No procedure-related complication was noted except one case of delayed cardiac tamponade due to the wire perforation. During the mean follow-up period of 1.9 ± 1.3 years, there was no adverse cardiac event, except one patient died of the complication of cardiac transplantation. The IVUS-guided wiringre-entry technique might be feasible and safe for the recanalization of complex CTO lesions. © 2018, Wiley Periodicals, Inc.

Phosphatidylinositol 4-Kinase IIIβ Is Required for Severe Acute Respiratory Syndrome Coronavirus Spike-mediated Cell Entry*

PubMed Central

Yang, Ning; Ma, Ping; Lang, Jianshe; Zhang, Yanli; Deng, Jiejie; Ju, Xiangwu; Zhang, Gongyi; Jiang, Chengyu

2012-01-01

Phosphatidylinositol kinases (PI kinases) play an important role in the life cycle of several viruses after infection. Using gene knockdown technology, we demonstrate that phosphatidylinositol 4-kinase IIIβ (PI4KB) is required for cellular entry by pseudoviruses bearing the severe acute respiratory syndrome-coronavirus (SARS-CoV) spike protein and that the cell entry mediated by SARS-CoV spike protein is strongly inhibited by knockdown of PI4KB. Consistent with this observation, pharmacological inhibitors of PI4KB blocked entry of SARS pseudovirions. Further research suggested that PI4P plays an essential role in SARS-CoV spike-mediated entry, which is regulated by the PI4P lipid microenvironment. We further demonstrate that PI4KB does not affect virus entry at the SARS-CoV S-ACE2 binding interface or at the stage of virus internalization but rather at or before virus fusion. Taken together, these results indicate a new function for PI4KB and suggest a new drug target for preventing SARS-CoV infection. PMID:22253445

Broad-spectrum antivirals against viral fusion

PubMed Central

Vigant, Frederic; Santos, Nuno C.; Lee, Benhur

2015-01-01

Effective antivirals have been developed against specific viruses, such as HIV, Hepatitis C virus and influenza virus. This ‘one bug–one drug’ approach to antiviral drug development can be successful, but it may be inadequate for responding to an increasing diversity of viruses that cause significant diseases in humans. The majority of viral pathogens that cause emerging and re-emerging infectious diseases are membrane-enveloped viruses, which require the fusion of viral and cell membranes for virus entry. Therefore, antivirals that target the membrane fusion process represent new paradigms for broad-spectrum antiviral discovery. In this Review, we discuss the mechanisms responsible for the fusion between virus and cell membranes and explore how broad-spectrum antivirals target this process to prevent virus entry. PMID:26075364

Risk Assessment During the Final Phase of an Uncontrolled Re-Entry

NASA Astrophysics Data System (ADS)

Gaudel, A.; Hourtolle, C.; Goester, J. F.; Fuentes, N.

2013-09-01

As French National Space Agency, CNES is empowered to monitor compliance with technical regulations of the French Space Operation Act, FSOA, and to take all necessary measures to ensure the safety of people, property, public health and environment for all space operations involving French responsibility at international level.Therefore, CNES developed ELECTRA that calculates the risk for ground population involved in three types of events: rocket launching, controlled re-entry and uncontrolled re-entry. For the first two cases, ELECTRA takes into account degraded cases due to a premature stop of propulsion.Major evolutions were implemented recently on ELECTRA to meet new users' requirements, like the risk assessment during the final phase of uncontrolled re-entry, that can be combined with the computed risk for each country involved by impacts.The purpose of this paper is to provide an overview of the ELECTRA method and main functionalities, and then to highlight these recent improvements.

Weighted re-randomization tests for minimization with unbalanced allocation.

PubMed

Han, Baoguang; Yu, Menggang; McEntegart, Damian

2013-01-01

Re-randomization test has been considered as a robust alternative to the traditional population model-based methods for analyzing randomized clinical trials. This is especially so when the clinical trials are randomized according to minimization, which is a popular covariate-adaptive randomization method for ensuring balance among prognostic factors. Among various re-randomization tests, fixed-entry-order re-randomization is advocated as an effective strategy when a temporal trend is suspected. Yet when the minimization is applied to trials with unequal allocation, fixed-entry-order re-randomization test is biased and thus compromised in power. We find that the bias is due to non-uniform re-allocation probabilities incurred by the re-randomization in this case. We therefore propose a weighted fixed-entry-order re-randomization test to overcome the bias. The performance of the new test was investigated in simulation studies that mimic the settings of a real clinical trial. The weighted re-randomization test was found to work well in the scenarios investigated including the presence of a strong temporal trend. Copyright © 2013 John Wiley & Sons, Ltd.

Re-refinement from deposited X-ray data can deliver improved models for most PDB entries

DOE Office of Scientific and Technical Information (OSTI.GOV)

Joosten, Robbie P.; Womack, Thomas; Vriend, Gert, E-mail: vriend@cmbi.ru.nl

2009-02-01

An evaluation of validation and real-space intervention possibilities for improving existing automated (re-)refinement methods. The deposition of X-ray data along with the customary structural models defining PDB entries makes it possible to apply large-scale re-refinement protocols to these entries, thus giving users the benefit of improvements in X-ray methods that have occurred since the structure was deposited. Automated gradient refinement is an effective method to achieve this goal, but real-space intervention is most often required in order to adequately address problems detected by structure-validation software. In order to improve the existing protocol, automated re-refinement was combined with structure validation andmore » difference-density peak analysis to produce a catalogue of problems in PDB entries that are amenable to automatic correction. It is shown that re-refinement can be effective in producing improvements, which are often associated with the systematic use of the TLS parameterization of B factors, even for relatively new and high-resolution PDB entries, while the accompanying manual or semi-manual map analysis and fitting steps show good prospects for eventual automation. It is proposed that the potential for simultaneous improvements in methods and in re-refinement results be further encouraged by broadening the scope of depositions to include refinement metadata and ultimately primary rather than reduced X-ray data.« less

Advances in Procedural Techniques - Antegrade

PubMed Central

Wilson, William; Spratt, James C.

2014-01-01

There have been many technological advances in antegrade CTO PCI, but perhaps most importantly has been the evolution of the “hybrid’ approach where ideally there exists a seamless interplay of antegrade wiring, antegrade dissection re-entry and retrograde approaches as dictated by procedural factors. Antegrade wire escalation with intimal tracking remains the preferred initial strategy in short CTOs without proximal cap ambiguity. More complex CTOs, however, usually require either a retrograde or an antegrade dissection re-entry approach, or both. Antegrade dissection re-entry is well suited to long occlusions where there is a healthy distal vessel and limited “interventional” collaterals. Early use of a dissection re-entry strategy will increase success rates, reduce complications, and minimise radiation exposure, contrast use as well as procedural times. Antegrade dissection can be achieved with a knuckle wire technique or the CrossBoss catheter whilst re-entry will be achieved in the most reproducible and reliable fashion by the Stingray balloon/wire. It should be avoided where there is potential for loss of large side branches. It remains to be seen whether use of newer dissection re-entry strategies will be associated with lower restenosis rates compared with the more uncontrolled subintimal tracking strategies such as STAR and whether stent insertion in the subintimal space is associated with higher rates of late stent malapposition and stent thrombosis. It is to be hoped that the algorithms, which have been developed to guide CTO operators, allow for a better transfer of knowledge and skills to increase uptake and acceptance of CTO PCI as a whole. PMID:24694104

Phosphatidylserine colocalizes with epichromatin in interphase nuclei and mitotic chromosomes

PubMed Central

Prudovsky, Igor; Vary, Calvin P.H.; Markaki, Yolanda; Olins, Ada L.; Olins, Donald E.

2012-01-01

Cycling eukaryotic cells rapidly re-establish the nuclear envelope and internal architecture following mitosis. Studies with a specific anti-nucleosome antibody recently demonstrated that the surface (“epichromatin”) of interphase and mitotic chromatin possesses a unique and conserved conformation, suggesting a role in postmitotic nuclear reformation. Here we present evidence showing that the anionic glycerophospholipid phosphatidylserine is specifically located in epichromatin throughout the cell cycle and is associated with nucleosome core histones. This suggests that chromatin bound phosphatidylserine may function as a nucleation site for the binding of ER and re-establishment of the nuclear envelope. PMID:22555604

34 CFR 668.4 - Payment period.

Code of Federal Regulations, 2011 CFR

2011-07-01

... the State; or (iii) Ten percent of the clock hours in the payment period. (f) Re-entry within 180 days... provisions of § 668.22. (g) Re-entry after 180 days or transfer. (1) Except as provided in paragraph (g)(3...

34 CFR 668.4 - Payment period.

Code of Federal Regulations, 2012 CFR

2012-07-01

... the State; or (iii) Ten percent of the clock hours in the payment period. (f) Re-entry within 180 days... provisions of § 668.22. (g) Re-entry after 180 days or transfer. (1) Except as provided in paragraph (g)(3...

34 CFR 668.4 - Payment period.

Code of Federal Regulations, 2013 CFR

2013-07-01

... the State; or (iii) Ten percent of the clock hours in the payment period. (f) Re-entry within 180 days... provisions of § 668.22. (g) Re-entry after 180 days or transfer. (1) Except as provided in paragraph (g)(3...

34 CFR 668.4 - Payment period.

Code of Federal Regulations, 2010 CFR

2010-07-01

... the State; or (iii) Ten percent of the clock hours in the payment period. (f) Re-entry within 180 days... provisions of § 668.22. (g) Re-entry after 180 days or transfer. (1) Except as provided in paragraph (g)(3...

34 CFR 668.4 - Payment period.

Code of Federal Regulations, 2014 CFR

2014-07-01

... the State; or (iii) Ten percent of the clock hours in the payment period. (f) Re-entry within 180 days... provisions of § 668.22. (g) Re-entry after 180 days or transfer. (1) Except as provided in paragraph (g)(3...

Protective Effects of the Launch/Entry Suit (LES) and the Liquid Cooling Garment(LCG) During Re-entry and Landing After Spaceflight

NASA Technical Reports Server (NTRS)

Perez, Sondra A.; Charles, John B.; Fortner, G. William; Hurst, Victor, IV; Meck, Janice V.

2002-01-01

Heart rate and arterial pressure were measured during shuttle re-entry, landing and initial standing in crewmembers with and without inflated anti-g suits and with and without liquid cooling garments (LCG). Preflight, three measurements were obtained seated, then standing. Prior to and during re-entry, arterial pressure and heart rate were measured every five minutes until wheels stop (WS). Then crewmembers initiated three seated and three standing measurements. In subjects without inflated anti-g suits, SBP and DBP were significantly lower during preflight standing (P = 0.006; P = 0.001 respectively) and at touchdown (TD) (P = 0.001; P = 0.003 respectively); standing SBP was significantly lower after WS. on-LeG users developed significantly higher heart rates during re-entry (P = 0.029, maxG; P = 0.05, TD; P = 0.02, post-WS seated; P = 0.01, post-WS standing) than LCG users. Our data suggest that the anti-g suit is effective, but the combined anti-g suit with LCG is more effective.

Inhibition of highly productive HIV-1 infection in T cells, primary human macrophages, microglia, and astrocytes by Sargassum fusiforme

PubMed Central

Paskaleva, Elena E; Lin, Xudong; Li, Wen; Cotter, Robin; Klein, Michael T; Roberge, Emily; Yu, Er K; Clark, Bruce; Veille, Jean-Claude; Liu, Yanze; Lee, David Y-W; Canki, Mario

2006-01-01

Background The high rate of HIV-1 mutation and increasing resistance to currently available antiretroviral (ART) therapies highlight the need for new antiviral agents. Products derived from natural sources have been shown to inhibit HIV-1 replication during various stages of the virus life cycle, and therefore represent a potential source of novel therapeutic agents. To expand our arsenal of therapeutics against HIV-1 infection, we investigated aqueous extract from Sargassum fusiforme (S. fusiforme) for ability to inhibit HIV-1 infection in the periphery, in T cells and human macrophages, and for ability to inhibit in the central nervous system (CNS), in microglia and astrocytes. Results S. fusiforme extract blocked HIV-1 infection and replication by over 90% in T cells, human macrophages and microglia, and it also inhibited pseudotyped HIV-1 (VSV/NL4-3) infection in human astrocytes by over 70%. Inhibition was mediated against both CXCR4 (X4) and CCR5 (R5)-tropic HIV-1, was dose dependant and long lasting, did not inhibit cell growth or viability, was not toxic to cells, and was comparable to inhibition by the nucleoside analogue 2', 3'-didoxycytidine (ddC). S. fusiforme treatment blocked direct cell-to-cell infection spread. To investigate at which point of the virus life cycle this inhibition occurs, we infected T cells and CD4-negative primary human astrocytes with HIV-1 pseudotyped with envelope glycoprotein of vesicular stomatitis virus (VSV), which bypasses the HIV receptor requirements. Infection by pseudotyped HIV-1 (VSV/NL4-3) was also inhibited in a dose dependant manner, although up to 57% less, as compared to inhibition of native NL4-3, indicating post-entry interferences. Conclusion This is the first report demonstrating S. fusiforme to be a potent inhibitor of highly productive HIV-1 infection and replication in T cells, in primary human macrophages, microglia, and astrocytes. Results with VSV/NL4-3 infection, suggest inhibition of both entry and post-entry events of the virus life cycle. Absence of cytotoxicity and high viability of treated cells also suggest that S. fusiforme is a potential source of novel naturally occurring antiretroviral compounds that inhibit HIV-1 infection and replication at more than one site of the virus life cycle. PMID:16725040

Small G Rac1 is involved in replication cycle of dengue serotype 2 virus in EAhy926 cells via the regulation of actin cytoskeleton.

PubMed

Zhang, Jing; Wu, Na; Gao, Na; Yan, Wenli; Sheng, Ziyang; Fan, Dongying; An, Jing

2016-05-01

Bleeding is a clinical characteristic of severe dengue and may be due to increased vascular permeability. However, the pathogenesis of severe dengue remains unclear. In this study, we showed that the Rac1-microfilament signal pathway was involved in the process of DENV serotype 2 (DENV2) infection in EAhy926 cells. DENV2 infection induced dynamic changes in actin organization, and treatment with Cytochalasin D or Jasplakinolide disrupted microfilament dynamics, reduced DENV2 entry, and inhibited DENV2 assembly and maturation. Rac1 activities decreased during the early phase and gradually increased by the late phase of infection. Expression of the dominant-negative form of Rac1 promoted DENV2 entry but inhibited viral assembly, maturation and release. Our findings demonstrated that Rac1 plays an important role in the DENV2 life cycle by regulating actin reorganization in EAhy926 cells. This finding provides further insight into the pathogenesis of severe dengue.

Vascular wall progenitor cells in health and disease.

PubMed

Psaltis, Peter J; Simari, Robert D

2015-04-10

The vasculature plays an indispensible role in organ development and maintenance of tissue homeostasis, such that disturbances to it impact greatly on developmental and postnatal health. Although cell turnover in healthy blood vessels is low, it increases considerably under pathological conditions. The principle sources for this phenomenon have long been considered to be the recruitment of cells from the peripheral circulation and the re-entry of mature cells in the vessel wall back into cell cycle. However, recent discoveries have also uncovered the presence of a range of multipotent and lineage-restricted progenitor cells in the mural layers of postnatal blood vessels, possessing high proliferative capacity and potential to generate endothelial, smooth muscle, hematopoietic or mesenchymal cell progeny. In particular, the tunica adventitia has emerged as a progenitor-rich compartment with niche-like characteristics that support and regulate vascular wall progenitor cells. Preliminary data indicate the involvement of some of these vascular wall progenitor cells in vascular disease states, adding weight to the notion that the adventitia is integral to vascular wall pathogenesis, and raising potential implications for clinical therapies. This review discusses the current body of evidence for the existence of vascular wall progenitor cell subpopulations from development to adulthood and addresses the gains made and significant challenges that lie ahead in trying to accurately delineate their identities, origins, regulatory pathways, and relevance to normal vascular structure and function, as well as disease. © 2015 American Heart Association, Inc.

The Potential of Computational Fluid Dynamics Simulation on Serial Monitoring of Hemodynamic Change in Type B Aortic Dissection

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yu, Simon C. H., E-mail: simonyu@cuhk.edu.hk; Liu, Wen; Wong, Randolph H. L.

PurposeWe aimed to assess the potential of computational fluid dynamics simulation (CFD) in detecting changes in pressure and flow velocity in response to morphological changes in type B aortic dissection.Materials and MethodsPressure and velocity in four morphological models of type B aortic dissection before and after closure of the entry tear were calculated with CFD and analyzed for changes among the different scenarios. The control model (Model 1) was patient specific and built from the DICOM data of CTA, which bore one entry tear and three re-entry tears. Models 2–4 were modifications of Model 1, with two re-entry tears lessmore » in Model 2, one re-entry tear more in Model 3, and a larger entry tear in Model 4.ResultsThe pressure and velocity pertaining to each of the morphological models were unique. Changes in pressure and velocity findings were accountable by the changes in morphological features of the different models. There was no blood flow in the false lumen across the entry tear after its closure, the blood flow direction across the re-entry tears was reversed after closure of the entry tear.ConclusionCFD simulation is probably useful to detect hemodynamic changes in the true and false lumens of type B aortic dissection in response to morphological changes, it may potentially be developed into a non-invasive and patient-specific tool for serial monitoring of hemodynamic changes of type B aortic dissection before and after treatment.« less

A mechanism underlying the effects of polyunsaturated fatty acids on breast cancer

PubMed Central

ZHANG, HAO; ZHOU, LEI; SHI, WEI; SONG, NING; YU, KARU; GU, YUCHUN

2012-01-01

Breast cancer is the most frequent cancer in women. Evidence suggests that the polyunsaturated fatty acids (PUFAs), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) affect breast cancer proliferation, differentiation and prognosis. However, the mechanism still remains unclear. In this study, the expression of transient receptor potential canonical (TRPC)3 was detected throughout the cell cytoplasm and at the cell surface of MCF-7 cells. Ca2+ entry was induced in these cells via activated TRPC3 by either the diacylglycerol analogue (OAG) or by intracellular Ca2+ store depletion. TRPC-mediated Ca2+ entry was inhibited by PUFAs including arachidonic acid (AA) and linolenic acid (LA) but not saturated fatty acids. Overexpression of the PUFA degradation enzyme, cyclooxygenase 2 (COX2), enhanced capacitative Ca2+ entry. In addition, inhibition of COX2 reduced [Ca2+]i. Nevertheless, inhibition of TRPC reduced the cell cycle S phase and cell migration, implicating a functional role for TRP-mediated Ca2+ entry in cell proliferation and invasion. Exogenous PUFA as well as a TRPC3 antagonist consistently attenuated breast cancer cell proliferation and migration, suggesting a mechanism in which PUFA restrains the breast cancer partly via its inhibition of TRPC channels. Additionally, our results also suggest that TRPC3 appears as a new mediator of breast cancer cell migration/invasion and represents a potential target for a new class of anticancer agent. PMID:22692672

Development and application of an empirical probability distribution for the prediction error of re-entry body maximum dynamic pressure

NASA Technical Reports Server (NTRS)

Lanzi, R. James; Vincent, Brett T.

1993-01-01

The relationship between actual and predicted re-entry maximum dynamic pressure is characterized using a probability density function and a cumulative distribution function derived from sounding rocket flight data. This paper explores the properties of this distribution and demonstrates applications of this data with observed sounding rocket re-entry body damage characteristics to assess probabilities of sustaining various levels of heating damage. The results from this paper effectively bridge the gap existing in sounding rocket reentry analysis between the known damage level/flight environment relationships and the predicted flight environment.

Childhood cancer survivors' school (re)entry: Australian parents' perceptions.

PubMed

McLoone, J K; Wakefield, C E; Cohn, R J

2013-07-01

Starting or returning to school after intense medical treatment can be academically and socially challenging for childhood cancer survivors. This study aimed to evaluate the school (re)entry experience of children who had recently completed cancer treatment. Forty-two semi-structured telephone interviews were conducted to explore parents' perceptions of their child's (re)entry to school after completing treatment (23 mothers, 19 fathers, parent mean age 39.5 years; child mean age 7.76 years). Interviews were analysed using the framework of Miles and Huberman and emergent themes were organised using QSR NVivo8. Parents closely monitored their child's school (re)entry and fostered close relationships with their child's teacher to ensure swift communication of concerns should they arise. The most commonly reported difficulty related to aspects of peer socialisation; survivors either displayed a limited understanding of social rules such as turn taking, or related more to older children or teachers relative to their peers. Additionally, parents placed a strong emphasis on their child's overall personal development, above academic achievement alone. Improved parent, clinician and teacher awareness of the importance of continued peer socialisation during the treatment period is recommended in order to limit the ongoing ramifications this may have on school (re)entry post-treatment completion. © 2013 John Wiley & Sons Ltd.

Variations in the quality of inpatient rehabilitation care to facilitate school re-entry and cognitive and communication function for children with TBI.

PubMed

Ennis, Stephanie K; Rivara, Frederick P; Mangione-Smith, Rita; Konodi, Mark A; Mackenzie, Ellen J; Jaffe, Kenneth M

2013-01-01

To examine variations in processes of paediatric inpatient rehabilitation care related to school re-entry and management of cognitive and communication impairments after traumatic brain injury. Retrospective cohort study. Adherence to care processes recommended for children (aged 0-17) with moderate-to-severe traumatic brain injury and admitted for inpatient rehabilitation was assessed. Quality-of-care indicators for processes supporting school re-entry and cognitive and communication rehabilitation were applied to measure variations in care delivered to 174 children across nine facilities using medical record review. Adherence rates (the number of times recommended care was delivered or attempted divided by the number of times care was indicated) were calculated, revealing substantial variations in care within and between facilities. Overall, children received 51.3% (95% CI = 31.9-70.7) and 72.3% (95% CI = 61.1-83.5), of the care recommended for school re-entry and cognitive and communication rehabilitation, respectively. Substantial variations exist in the delivery of paediatric inpatient rehabilitation care processes for managing school re-entry and cognitive and communication impairments after traumatic brain injury. Measures of association of these care processes with patient outcomes are necessary. Reduction in this variation is essential to improving quality of care.

Re-refinement from deposited X-ray data can deliver improved models for most PDB entries.

PubMed

Joosten, Robbie P; Womack, Thomas; Vriend, Gert; Bricogne, Gérard

2009-02-01

The deposition of X-ray data along with the customary structural models defining PDB entries makes it possible to apply large-scale re-refinement protocols to these entries, thus giving users the benefit of improvements in X-ray methods that have occurred since the structure was deposited. Automated gradient refinement is an effective method to achieve this goal, but real-space intervention is most often required in order to adequately address problems detected by structure-validation software. In order to improve the existing protocol, automated re-refinement was combined with structure validation and difference-density peak analysis to produce a catalogue of problems in PDB entries that are amenable to automatic correction. It is shown that re-refinement can be effective in producing improvements, which are often associated with the systematic use of the TLS parameterization of B factors, even for relatively new and high-resolution PDB entries, while the accompanying manual or semi-manual map analysis and fitting steps show good prospects for eventual automation. It is proposed that the potential for simultaneous improvements in methods and in re-refinement results be further encouraged by broadening the scope of depositions to include refinement metadata and ultimately primary rather than reduced X-ray data.

Base flow investigation of the Apollo AS-202 Command Module

NASA Astrophysics Data System (ADS)

Walpot, Louis M. G.; Wright, Michael J.; Noeding, Peter; Schrijer, Ferry

2012-01-01

A major contributor to the overall vehicle mass of re-entry vehicles is the afterbody thermal protection system. This is due to the large acreage (equal or bigger than that of the forebody) to be protected. The present predictive capabilities for base flows are comparatively lower than those for windward flowfields and offer therefore a substantial potential for improving the design of future re-entry vehicles. To that end, it is essential to address the accuracy of high fidelity CFD tools exercised in the US and EU, which motivates a thorough investigation of the present status of hypersonic flight afterbody heating. This paper addresses the predictive capabilities of afterbody flow fields of re-entry vehicles investigated in the frame of the NATO/RTO-RTG-043 task group. First, the verification of base flow topologies on the basis of available wind-tunnel results performed under controlled supersonic conditions (i.e. cold flows devoid of reactive effects) is performed. Such tests address the detailed characterization of the base flow with particular emphasis on separation/reattachment and their relation to Mach number effects. The tests have been performed on an Apollo-like re-entry capsule configuration. Second, the tools validated in the frame of the previous effort are exercised and appraised against flight-test data collected during the Apollo AS-202 re-entry.

EntrySat: A 3U CubeStat to study the reentry atmospheric environment

NASA Astrophysics Data System (ADS)

Anthony, Sournac; Raphael, Garcia; David, Mimoun; Jeremie, Chaix

2016-04-01

ISAE France Entrysat has for main scientific objective the study of uncontrolled atmospheric re-entry. This project, is developed by ISAE in collaboration with ONERA and University of Toulouse, is funded by CNES, in the overall frame of the QB50 project. This nano-satellite is a 3U Cubesat measuring 34*10*10 cm3, similar to secondary debris produced during the break up of a spacecraft. EntrySat will collect the external and internal temperatures, pressure, heat flux, attitude variations and drag force of the satellite between ≈150 and 90 km before its destruction in the atmosphere, and transmit them during the re-entry using the IRIDIUM satellite network. The result will be compared with the computations of MUSIC/FAST, a new 6-degree of freedom code developed by ONERA to predict the trajectory of space debris. In order to fulfil the scientific objectives, the satellite will acquire 18 re-entry sensors signals, convert them and compress them, thanks to an electronic board developed by ISAE students in cooperation with EREMS. In order to transmit these data every second during the re-entry phase, the satellite will use an IRIDIUM connection. In order to keep a stable enough attitudes during this phase, a simple attitude orbit and control system using magnetotorquers and an inertial measurement unit (IMU) is developed at ISAE by students. A commercial GPS board is also integrated in the satellite into Entry Sat to determine its position and velocity which are necessary during the re-entry phase. This GPS will also be used to synchronize the on-board clock with the real-time UTC data. During the orbital phase (≈2 year) EntrySat measurements will be recorded transmitted through a more classical "UHF/VHF" connection. Preference for presentation: Poster Most suitable session: Author for correspondence: Dr Raphael F. Garcia ISAE 10, ave E. Belin, 31400 Toulouse, France Raphael.GARCIA@isae.fr +33 5 61 33 81 14

Improving cardiac conduction with a skeletal muscle sodium channel by gene and cell therapy

PubMed Central

Lu, Jia; Wang, Hong-Zhan; Jia, Zhiheng; Zuckerman, Joan; Lu, Zhongju; Guo, Yuanjian; Boink, Gerard J.J.; Brink, Peter R.; Robinson, Richard B.; Entcheva, Emilia; Cohen, Ira S.

2012-01-01

The voltage-gated Na+ channel is a critical determinant of the action potential upstroke. Increasing Na+ conductance may speed action potential propagation. Here we propose use of the skeletal muscle Na+ channel SkM1 as a more favorable gene than the cardiac isoform SCN5A to enhance conduction velocity in depolarized cardiac tissue. We used cells which electrically coupled with cardiac myocytes as a delivery platform to introduce the Na+ channels. HEK293 cells were stably transfected with SkM1 or SCN5A. SkM1 had a more depolarized (18mV shift) inactivation curve than SCN5A. We also found that SkM1 recovered faster from inactivation than SCN5A. When coupled with SkM1 expressing cells, cultured myocytes showed an increase in the dV/dtmax of the action potential. Expression of SCN5A had no such effect. In an in vitro cardiac syncytium, coculture of neonatal cardiac myocytes with SkM1 expressing but not SCN5A expressing cells significantly increased the conduction velocity under both normal and depolarized conditions. In an in vitro re-entry model induced by high frequency stimulation, expression of SkM1 also enhanced angular velocity of the induced re-entry. These results suggest that cells carrying a Na+ channel with a more depolarized inactivation curve can improve cardiac excitability and conduction in depolarized tissues. PMID:22526298

Cell-autonomous defense, re-organization and trafficking of membranes in plant-microbe interactions.

PubMed

Dörmann, Peter; Kim, Hyeran; Ott, Thomas; Schulze-Lefert, Paul; Trujillo, Marco; Wewer, Vera; Hückelhoven, Ralph

2014-12-01

Plant cells dynamically change their architecture and molecular composition following encounters with beneficial or parasitic microbes, a process referred to as host cell reprogramming. Cell-autonomous defense reactions are typically polarized to the plant cell periphery underneath microbial contact sites, including de novo cell wall biosynthesis. Alternatively, host cell reprogramming converges in the biogenesis of membrane-enveloped compartments for accommodation of beneficial bacteria or invasive infection structures of filamentous microbes. Recent advances have revealed that, in response to microbial encounters, plasma membrane symmetry is broken, membrane tethering and SNARE complexes are recruited, lipid composition changes and plasma membrane-to-cytoskeleton signaling is activated, either for pre-invasive defense or for microbial entry. We provide a critical appraisal on recent studies with a focus on how plant cells re-structure membranes and the associated cytoskeleton in interactions with microbial pathogens, nitrogen-fixing rhizobia and mycorrhiza fungi. © 2014 The Authors. New Phytologist © 2014 New Phytologist Trust.

Kinetics of Neuraminidase Action on Glycoproteins by One- and Two-Dimensional NMR

ERIC Educational Resources Information Center

Barb, Adam W.; Glushka, John N.; Prestegard, James H.

2011-01-01

The surfaces of mammalian cells are coated with complex carbohydrates, many terminated with a negatively charged "N"-acetylneuraminic acid residue. This motif is specifically targeted by pathogens, including influenza viruses and many pathogenic bacteria, to gain entry into the cell. A necessary step in the influenza virus life cycle is the…

Molecular mechanisms involved in the early steps of flavivirus cell entry.

PubMed

Kaufmann, Bärbel; Rossmann, Michael G

2011-01-01

Flaviviruses enter their host cells by receptor-mediated endocytosis, a well-orchestrated process of receptor recognition, penetration and uncoating. Recent findings on these early steps in the life cycle of flaviviruses are the focus of this review. Copyright © 2010 Institut Pasteur. Published by Elsevier SAS. All rights reserved.

Pre-exposure to 50 Hz-electromagnetic fields enhanced the antiproliferative efficacy of 5-fluorouracil in breast cancer MCF-7 cells

PubMed Central

Chen, Sha; Sun, Xiongshan; Guan, Xiao; Yang, Yao; Peng, Bingjie; Pan, Xiaodong; Li, Jinfang; Yi, Weijing; Li, Peng; Zhang, Hongwei; Feng, Dongfang; Chen, An; Li, Xiaohui; Yin, Zuoming

2018-01-01

Resistance to 5-fluorouracil (5-FU) and its induced immune suppression have prevented its extensive application in the clinical treatment of breast cancer. In this study, the combined effect of 50 Hz-EMFs and 5-FU in the treatment of breast cancer was explored. MCF-7 and MCF10A cells were pre-exposed to 50 Hz-EMFs for 0, 2, 4, 8 and 12 h and then treated with different concentrations of 5-FU for 24 h; cell viability was analyzed by MTT assay and flow cytometry. After pre-exposure to 50 Hz-EMFs for 12 h, apoptosis and cell cycle distribution in MCF-7 and MCF10A cells were detected via flow cytometry and DNA synthesis was measured by EdU incorporation assay. Apoptosis-related and cell cycle-related gene and protein expression levels were monitored by qPCR and western blotting. Pre-exposure to 50 Hz-EMFs for 12 h enhanced the antiproliferative effect of 5-FU in breast cancer cell line MCF-7 in a dose-dependent manner but not in normal human breast epithelial cell line MCF10A. Exposure to 50 Hz-EMFs had no effect on apoptosis and P53 expression of MCF-7 and MCF10A cells, whereas it promoted DNA synthesis, induced entry of MCF-7 cells into the S phase of cell cycle, and upregulated the expression levels of cell cycle-related proteins Cyclin D1 and Cyclin E. Considering the pharmacological mechanisms of 5-FU in specifically disrupting DNA synthesis, this enhanced inhibitory effect might have resulted from the specific sensitivity of MCF7 cells in active S phase to 5-FU. Our findings demonstrate the enhanced cytotoxic activity of 5-FU on MCF7 cells through promoting entry into the S phase of the cell cycle via exposure to 50 Hz-EMFs, which provides a novel method of cancer treatment based on the combinatorial use of 50 Hz-EMFs and chemotherapy. PMID:29617363

An ex vivo continuous passive motion model in a porcine knee for assessing primary stability of cell-free collagen gel plugs.

PubMed

Efe, Turgay; Schofer, Markus D; Füglein, Alexander; Timmesfeld, Nina; Fuchs-Winkelmann, Susanne; Stein, Thomas; El-Zayat, Bilal Farouk; Paletta, Jürgen Rj; Heyse, Thomas J

2010-12-15

Primary stability of cartilage repair constructs is of the utmost importance in the clinical setting but few continuous passive motion (CPM) models are available. Our study aimed to establish a novel ex vivo CPM animal model and to evaluate the required motion cycles for testing the mechanical properties of a new cell-free collagen type I gel plug (CaReS®-1S). A novel ex vivo CPM device was developed. Full-thickness cartilage defects (11 mm diameter by 6 mm deep) were created on the medial femoral condyle of porcine knee specimens. CaReS®-1S was implanted in 16 animals and each knee underwent continuous passive motion. After 0, 2000, 4000, 6000, and 8000 motions, standardized digital pictures of the grafts were taken, focusing on the worn surfaces. The percentage of worn surface on the total CaReS®-1S surface was evaluated with image processing software. Significant differences in the worn surface were recorded between 0 and 2000 motion cycles (p < 0.0001). After 2000 motion cycles, there was no significant difference. No total delamination of CaReS®-1S with an empty defect site was recorded. The ex vivo CPM animal model is appropriate in investigating CaReS®-1S durability under continuous passive motion. 2000 motion cycles appear adequate to assess the primary stability of type I collagen gels used to repair focal chondral defects.

An ex vivo continuous passive motion model in a porcine knee for assessing primary stability of cell-free collagen gel plugs

PubMed Central

2010-01-01

Background Primary stability of cartilage repair constructs is of the utmost importance in the clinical setting but few continuous passive motion (CPM) models are available. Our study aimed to establish a novel ex vivo CPM animal model and to evaluate the required motion cycles for testing the mechanical properties of a new cell-free collagen type I gel plug (CaReS®-1S). Methods A novel ex vivo CPM device was developed. Full-thickness cartilage defects (11 mm diameter by 6 mm deep) were created on the medial femoral condyle of porcine knee specimens. CaReS®-1S was implanted in 16 animals and each knee underwent continuous passive motion. After 0, 2000, 4000, 6000, and 8000 motions, standardized digital pictures of the grafts were taken, focusing on the worn surfaces. The percentage of worn surface on the total CaReS®-1S surface was evaluated with image processing software. Results Significant differences in the worn surface were recorded between 0 and 2000 motion cycles (p < 0.0001). After 2000 motion cycles, there was no significant difference. No total delamination of CaReS®-1S with an empty defect site was recorded. Conclusion The ex vivo CPM animal model is appropriate in investigating CaReS®-1S durability under continuous passive motion. 2000 motion cycles appear adequate to assess the primary stability of type I collagen gels used to repair focal chondral defects. PMID:21159196

Hypersonic Cruise and Re-Entry Radio Frequency Blackout Mitigation: Alleviating the Communications Blackout Problem

NASA Technical Reports Server (NTRS)

Manning, Robert M.

2017-01-01

The work presented here will be a review of a NASA effort to provide a method to transmit and receive RF communications and telemetry through a re-entry plasma thus alleviating the classical RF blackout phenomenon.

The Role of Counselling and Parental Encouragement on Re-Entry of Adolescents into Secondary Schools in Abia State, Nigeria

ERIC Educational Resources Information Center

Alika, Henrietta Ijeoma; Ohanaka, Blessing Ijeoma

2013-01-01

This paper examined the role of counselling, and parental encouragement on re-entry of adolescents into secondary school in Abia State, Nigeria. A total of 353 adolescents who re-entered school were selected from six secondary schools in the State through a simple random sampling technique. A validated questionnaire was used for data analysis.…

Vocational Interest as a Correlate of Re-Entry of Girls into School in Edo State, Nigeria: Implications for Counselling

ERIC Educational Resources Information Center

Alika, Ijeoma Henrietta; Egbochuku, Elizabeth Omotunde

2012-01-01

The study investigated the relationship between vocational interest socio-economic status and re-entry of girls into school in Edo State. The research design adopted was correlational because it sought to establish the relationship between the independent variable and the dependent variable. A sample size of 306 girls who re-enrolled in institutes…

TESTIN Induces Rapid Death and Suppresses Proliferation in Childhood B Acute Lymphoblastic Leukaemia Cells

PubMed Central

Weeks, Robert J.; Ludgate, Jackie L.; LeMée, Gwenn; Morison, Ian M.

2016-01-01

Background Childhood acute lymphoblastic leukaemia (ALL) is the most common malignancy in children. Despite high cure rates, side effects and late consequences of the intensive treatments are common. Unquestionably, the identification of new therapeutic targets will lead to safer, more effective treatments. We identified TES promoter methylation and transcriptional silencing as a very common molecular abnormality in childhood ALL, irrespective of molecular subtype. The aims of the present study were to demonstrate that TES promoter methylation is aberrant, to determine the effects of TES re-expression in ALL, and to determine if those effects are mediated via TP53 activity. Methods Normal fetal and adult tissue DNA was isolated and TES promoter methylation determined by Sequenom MassARRAY. Quantitative RT-PCR and immunoblot were used to confirm re-expression of TES in ALL cell lines after 5’-aza-2’-deoxycytidine (decitabine) exposure or transfection with TES expression plasmids. The effects of TES re-expression on ALL cells were investigated using standard cell proliferation, cell death and cell cycle assays. Results In this study, we confirm that the TES promoter is unmethylated in normal adult and fetal tissues. We report that decitabine treatment of ALL cell lines results in demethylation of the TES promoter and attendant expression of TES mRNA. Re-expression of TESTIN protein in ALL cells using expression plasmid transfection results in rapid cell death or cell cycle arrest independent of TP53 activity. Conclusions These results suggest that TES is aberrantly methylated in ALL and that re-expression of TESTIN has anti-leukaemia effects which point to novel therapeutic opportunities for childhood ALL. PMID:26985820

DNA Damage and Genomic Instability Induced by Inappropriate DNA Re-replication

DTIC Science & Technology

2007-04-01

Conway, A., Lockhart, D. J., Davis, R. W., Brewer , B. J., and Fangman, W. L. (2001). Replication dynamics of the yeast genome. Science 294, 115–121... Brewer , B. J. (2001). An origin-deficient yeast artificial chromosome triggers a cell cycle checkpoint. Mol. Cell 7, 705–713. Vas, A., Mok, W., and...replication in yeast cells. We have demonstrated that re-replication induces a rapid and significant decrease in cell viability and a cellular DNA damage

Dissection and re-entry techniques and longer-term outcomes following successful percutaneous coronary intervention of chronic total occlusion.

PubMed

Rinfret, Stéphane; Ribeiro, Henrique Barbosa; Nguyen, Can Manh; Nombela-Franco, Luis; Ureña, Marina; Rodés-Cabau, Josep

2014-11-01

New techniques involving dissection of the subintimal space and re-entry into the true lumen increase success rates in chronic total occlusion (CTO) percutaneous coronary intervention (PCI). However, their long-term safety and efficacy were unknown. This study included a series of consecutive patients who underwent CTO PCI. All patients who did not present events were contacted 12 to 18 months after their PCI. The combined incidence of cardiac death, myocardial infarction, ischemia-driven target-vessel revascularization (TVR), or reocclusion was assessed as our primary outcome. From January 2010 to January 2013, of 212 CTOs treated in our CTO program, 192 (91%) were successfully opened (in 179 patients). Follow-up data were available for 187 CTOs (97.4%), with 82 (44%) that were opened with dissection re-entry and 105 (56%) with conventional wire escalation techniques. At a median follow-up of 398 days, the primary outcome occurred in 18 of 179 CTOs treated (10.7%), driven by TVR. No patient died from cardiac causes. Eleven CTOs (15.2%) treated with dissection re-entry versus 7 CTOs (7.3%) treated with wire escalation presented with the primary outcome (p = 0.17). With multivariate adjustment, dissection re-entry techniques had no significant impact on outcomes. However, treatment of an in-stent occlusion was independently associated with TVR (hazards ratio >6.0, p <0.001). In conclusion, dissection re-entry techniques have minimal impact on long-term outcomes after CTO PCI, which are favorable in most patients. However, treatment of an in-stent occlusion and use of sirolimus-eluting stent were predictors of subsequent adverse outcomes. Copyright © 2014 Elsevier Inc. All rights reserved.

Role of Re-entry Tears on the Dynamics of Type B Dissection Flap.

PubMed

Canchi, Saranya; Guo, Xiaomei; Phillips, Matt; Berwick, Zachary; Kratzberg, Jarin; Krieger, Joshua; Roeder, Blayne; Haulon, Stephan; Chambers, Sean; Kassab, Ghassan S

2018-01-01

Mortality during follow-up after acute Type B aortic dissection is substantial with aortic expansion observed in over 59% of the patients. Lumen pressure differential is considered a prime contributing factor for aortic dilation after propagation. The objective of the study was to evaluate the relationship between changes in vessel geometry with and without lumen pressure differential post propagation in an ex vivo porcine model with comparison with patient clinical data. A pulse duplicator system was utilized to propagate the dissection within descending thoracic porcine aortic vessels for set proximal (%circumference of the entry tear: 40%, axial length: 2 cm) and re-entry (50% of distal vessel circumference) tear geometry. Measurements of lumen pressure differential were made along with quantification of vessel geometry (n = 16). The magnitude of mean lumen pressure difference measured after propagation was low (~ 5 mmHg) with higher pressures measured in false lumen and as anticipated the pressure difference approached zero after the creation of distal re-entry tear. False lumen Dissection Ratio (FDR) defined as arc length of dissected wall divided by arc length of dissection flap, had mean value of 1.59 ± 0.01 at pressure of 120/80 mmHg post propagation with increasing values with increase in pulse pressure that was not rescued with the creation of distal re-entry tear (p < 0.01). An average FDR of 1.87 ± 0.27 was measured in patients with acute Type B dissection. Higher FDR value (FDR = 1 implies zero dissection) in the presence of distal re-entry tear demonstrates an acute change in vessel morphology in response to the dissection independent of local pressure changes challenges the re-apposition of the aortic wall.

FLPP IXV Re-entry Vehicle, Transonic Characterisation Based on FOI T1500 Wind Tunnel Tests and CFD

NASA Astrophysics Data System (ADS)

Torngren, L.; Chiarelli, C.; Mareschi, V.; Tribot, J.-P.; Binetti, P.; Walloschek, T.

2009-01-01

The European Space Agency ESA, has engaged in 2004, the IXV project (Intermediate eXperimental Vehicle) which is part of the FLPP (Future Launcher Preparatory Programme) aiming at answering to critical technological issues, while supporting the future generation launchers and to improve in general European capabilities in the strategic field of atmospheric re-entry for space transportation, exploration and scientific applications. The IXV key mission and system objectives are the design, development, manufacturing, assembling and on-ground to in-flight verification of an autonomous European lifting and aerodynamically controlled re-entry system, integrating the critical re-entry technologies at the system level. The current IXV vehicle is a slender body type exhibiting rounded shape, thick body controlled by means of two control surfaces. The current mission is to perform an atmospheric re- entry ended by a safe recovery in supersonic regime. A potential extension of the flight domain down to the transonic regime was proposed to be analyzed. The objectives were to study the capability of the IXV for flying autonomously enabling a recovery of the vehicle by means of a subsonic parachute based DRS. The vehicle designed for the hypersonic speeds integrating a large base with only two control surfaces located close to the plane of symmetry is definitively not tuned for transonic ones. CFD done by Thales Alenia Space and wind tunnel activities involving FOI T1500 facility contributed to built up an Aerodynamic Data Base (AEDB) to be used as inputs for flying qualities analysis and re-entry simulations. The paper presents the main objectives of the transonic activities with emphasis on CFD and WTT including a description of the different prediction tools and discussing the main outcomes of the current data comparisons.

DYRK1A Is a Regulator of S-Phase Entry in Hepatic Progenitor Cells.

PubMed

Kruitwagen, Hedwig S; Westendorp, Bart; Viebahn, Cornelia S; Post, Krista; van Wolferen, Monique E; Oosterhoff, Loes A; Egan, David A; Delabar, Jean-Maurice; Toussaint, Mathilda J; Schotanus, Baukje A; de Bruin, Alain; Rothuizen, Jan; Penning, Louis C; Spee, Bart

2018-01-15

Hepatic progenitor cells (HPCs) are adult liver stem cells that act as second line of defense in liver regeneration. They are normally quiescent, but in case of severe liver damage, HPC proliferation is triggered by external activation mechanisms from their niche. Although several important proproliferative mechanisms have been described, it is not known which key intracellular regulators govern the switch between HPC quiescence and active cell cycle. We performed a high-throughput kinome small interfering RNA (siRNA) screen in HepaRG cells, a HPC-like cell line, and evaluated the effect on proliferation with a 5-ethynyl-2'-deoxyuridine (EdU) incorporation assay. One hit increased the percentage of EdU-positive cells after knockdown: dual specificity tyrosine phosphorylation regulated kinase 1A (DYRK1A). Although upon DYRK1A silencing, the percentage of EdU- and phosphorylated histone H3 (pH3)-positive cells was increased, and total cell numbers were not increased, possibly through a subsequent delay in cell cycle progression. This phenotype was confirmed with chemical inhibition of DYRK1A using harmine and with primary HPCs cultured as liver organoids. DYRK1A inhibition impaired Dimerization Partner, RB-like, E2F, and multivulva class B (DREAM) complex formation in HPCs and abolished its transcriptional repression on cell cycle progression. To further analyze DYRK1A function in HPC proliferation, liver organoid cultures were established from mBACtgDyrk1A mice, which harbor one extra copy of the murine Dyrk1a gene (Dyrk+++). Dyrk+++ organoids had both a reduced percentage of EdU-positive cells and reduced proliferation compared with wild-type organoids. This study provides evidence for an essential role of DYRK1A as balanced regulator of S-phase entry in HPCs. An exact gene dosage is crucial, as both DYRK1A deficiency and overexpression affect HPC cell cycle progression.

Regulation of store-operated Ca{sup 2+} entry activity by cell cycle dependent up-regulation of Orai2 in brain capillary endothelial cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kito, Hiroaki; Department of Pharmacology, Division of Pathological Sciences, Kyoto Pharmaceutical University, Kyoto; Yamamura, Hisao

2015-04-10

Store-operated Ca{sup 2+} entry (SOCE) via Orai1 and STIM1 complex is supposed to have obligatory roles in the regulation of cellular functions of vascular endothelial cells, while little is known about the contribution of Orai2. Quantitative PCR and Western blot analyses indicated the expression of Orai2 and STIM2, in addition to Orai1 and STIM1 in bovine brain capillary endothelial cell line, t-BBEC117. During the exponential growth of t-BBEC117, the knockdown of Orai1 and STIM1 significantly reduced the SOCE activity, whereas Orai2 and STIM2 siRNAs had no effect. To examine whether endogenous SOCE activity contributes to the regulation of cell cyclemore » progression, t-BBEC117 were synchronized using double thymidine blockage. At the G2/M phase, Ca{sup 2+} influx via SOCE was decreased and Orai2 expression was increased compared to the G0/G1 phase. When Orai2 was knocked down at the G2/M phase, the decrease in SOCE was removed, and cell proliferation was partly attenuated. Taken together, Orai1 significantly contributes to cell proliferation via the functional expression, which is presumably independent of the cell cycle phases. In construct, Orai2 is specifically up-regulated during the G2/M phase, negatively modulates the SOCE activity, and may contribute to the regulation of cell cycle progression in brain capillary endothelial cells. - Highlights: • Orai1 is essential for SOCE activity in brain capillary endothelial cells (BCECs). • Cell cycle independent expression of Orai1 regulated SOCE and cell proliferation. • Orai2 was up-regulated only at G2/M phase and this consequently reduced SOCE. • Orai2 as well as Orai1 is a key player controlling SOCE and proliferation in BCECs.« less

VDAC3 and Mps1 negatively regulate ciliogenesis.

PubMed

Majumder, Shubhra; Fisk, Harold A

2013-03-01

Centrosomes serve to organize new centrioles in cycling cells, whereas in quiescent cells they assemble primary cilia. We have recently shown that the mitochondrial porin VDAC3 is also a centrosomal protein that is predominantly associated with the mother centriole and modulates centriole assembly by recruiting Mps1 to centrosomes. Here, we show that depletion of VDAC3 causes inappropriate ciliogenesis in cycling cells, while expression of GFP-VDAC3 suppresses ciliogenesis in quiescent cells. Mps1 also negatively regulates ciliogenesis, and the inappropriate ciliogenesis caused by VDAC3 depletion can be bypassed by targeting Mps1 to centrosomes independently of VDAC3. Thus, our data show that a VDAC3-Mps1 module at the centrosome promotes ciliary disassembly during cell cycle entry and suppresses cilia assembly in proliferating cells. Our data also suggests that VDAC3 might be a link between mitochondrial dysfunction and ciliopathies in mammalian cells.

Chronological Lifespan in Yeast Is Dependent on the Accumulation of Storage Carbohydrates Mediated by Yak1, Mck1 and Rim15 Kinases

PubMed Central

Tang, Yingzhi; Quan, Zhenzhen; Zhang, Zhe; Oliver, Stephen G.; Zhang, Nianshu

2016-01-01

Upon starvation for glucose or any other macronutrient, yeast cells exit from the mitotic cell cycle and acquire a set of characteristics that are specific to quiescent cells to ensure longevity. Little is known about the molecular determinants that orchestrate quiescence entry and lifespan extension. Using starvation-specific gene reporters, we screened a subset of the yeast deletion library representing the genes encoding ‘signaling’ proteins. Apart from the previously characterised Rim15, Mck1 and Yak1 kinases, the SNF1/AMPK complex, the cell wall integrity pathway and a number of cell cycle regulators were shown to be necessary for proper quiescence establishment and for extension of chronological lifespan (CLS), suggesting that entry into quiescence requires the integration of starvation signals transmitted via multiple signaling pathways. The CLS of these signaling mutants, and those of the single, double and triple mutants of RIM15, YAK1 and MCK1 correlates well with the amount of storage carbohydrates but poorly with transition-phase cell cycle status. Combined removal of the glycogen and trehalose biosynthetic genes, especially GSY2 and TPS1, nearly abolishes the accumulation of storage carbohydrates and severely reduces CLS. Concurrent overexpression of GSY2 and TSL1 or supplementation of trehalose to the growth medium ameliorates the severe CLS defects displayed by the signaling mutants (rim15Δyak1Δ or rim15Δmck1Δ). Furthermore, we reveal that the levels of intracellular reactive oxygen species are cooperatively controlled by Yak1, Rim15 and Mck1, and the three kinases mediate the TOR1-regulated accumulation of storage carbohydrates and CLS extension. Our data support the hypothesis that metabolic reprogramming to accumulate energy stores and the activation of anti-oxidant defence systems are coordinated by Yak1, Rim15 and Mck1 kinases to ensure quiescence entry and lifespan extension in yeast. PMID:27923067

Chronological Lifespan in Yeast Is Dependent on the Accumulation of Storage Carbohydrates Mediated by Yak1, Mck1 and Rim15 Kinases.

PubMed

Cao, Lu; Tang, Yingzhi; Quan, Zhenzhen; Zhang, Zhe; Oliver, Stephen G; Zhang, Nianshu

2016-12-01

Upon starvation for glucose or any other macronutrient, yeast cells exit from the mitotic cell cycle and acquire a set of characteristics that are specific to quiescent cells to ensure longevity. Little is known about the molecular determinants that orchestrate quiescence entry and lifespan extension. Using starvation-specific gene reporters, we screened a subset of the yeast deletion library representing the genes encoding 'signaling' proteins. Apart from the previously characterised Rim15, Mck1 and Yak1 kinases, the SNF1/AMPK complex, the cell wall integrity pathway and a number of cell cycle regulators were shown to be necessary for proper quiescence establishment and for extension of chronological lifespan (CLS), suggesting that entry into quiescence requires the integration of starvation signals transmitted via multiple signaling pathways. The CLS of these signaling mutants, and those of the single, double and triple mutants of RIM15, YAK1 and MCK1 correlates well with the amount of storage carbohydrates but poorly with transition-phase cell cycle status. Combined removal of the glycogen and trehalose biosynthetic genes, especially GSY2 and TPS1, nearly abolishes the accumulation of storage carbohydrates and severely reduces CLS. Concurrent overexpression of GSY2 and TSL1 or supplementation of trehalose to the growth medium ameliorates the severe CLS defects displayed by the signaling mutants (rim15Δyak1Δ or rim15Δmck1Δ). Furthermore, we reveal that the levels of intracellular reactive oxygen species are cooperatively controlled by Yak1, Rim15 and Mck1, and the three kinases mediate the TOR1-regulated accumulation of storage carbohydrates and CLS extension. Our data support the hypothesis that metabolic reprogramming to accumulate energy stores and the activation of anti-oxidant defence systems are coordinated by Yak1, Rim15 and Mck1 kinases to ensure quiescence entry and lifespan extension in yeast.

Anaphase-Promoting Complex/Cyclosome-Cdh1-Mediated Proteolysis of the Forkhead Box M1 Transcription Factor Is Critical for Regulated Entry into S Phase▿

PubMed Central

Park, Hyun Jung; Costa, Robert H.; Lau, Lester F.; Tyner, Angela L.; Raychaudhuri, Pradip

2008-01-01

The forkhead box M1 (FoxM1) transcription factor is overexpressed in many cancers, and in mouse models it is required for tumor progression. FoxM1 activates expression of the cell cycle genes required for both S and M phase progression. Here we demonstrate that FoxM1 is degraded in late mitosis and early G1 phase by the anaphase-promoting complex/cyclosome (APC/C) E3 ubiquitin ligase. FoxM1 interacts with the APC/C complex and its adaptor, Cdh1. Expression of Cdh1 stimulated degradation of the FoxM1 protein, and depletion of Cdh1 resulted in stabilization of the FoxM1 protein in late mitosis and in early G1 phase of the cell cycle. Cdh1 has been implicated in regulating S phase entry. We show that codepletion of FoxM1 inhibits early S phase entry observed in Cdh1-depleted cells. The N-terminal region of FoxM1 contains both destruction box (D box) and KEN box sequences that are required for targeting by Cdh1. Mutation of either the D box sequence or the KEN box sequence stabilized FoxM1 and blocked Cdh1-induced proteolysis. Cells expressing a nondegradable form of FoxM1 entered S phase rapidly following release from M phase arrest. Together, our observations show that FoxM1 is one of the targets of Cdh1 in late M or early G1 phase and that its proteolysis is important for regulated entry into S phase. PMID:18573889

Anaphase-promoting complex/cyclosome-CDH1-mediated proteolysis of the forkhead box M1 transcription factor is critical for regulated entry into S phase.

PubMed

Park, Hyun Jung; Costa, Robert H; Lau, Lester F; Tyner, Angela L; Raychaudhuri, Pradip

2008-09-01

The forkhead box M1 (FoxM1) transcription factor is overexpressed in many cancers, and in mouse models it is required for tumor progression. FoxM1 activates expression of the cell cycle genes required for both S and M phase progression. Here we demonstrate that FoxM1 is degraded in late mitosis and early G(1) phase by the anaphase-promoting complex/cyclosome (APC/C) E3 ubiquitin ligase. FoxM1 interacts with the APC/C complex and its adaptor, Cdh1. Expression of Cdh1 stimulated degradation of the FoxM1 protein, and depletion of Cdh1 resulted in stabilization of the FoxM1 protein in late mitosis and in early G(1) phase of the cell cycle. Cdh1 has been implicated in regulating S phase entry. We show that codepletion of FoxM1 inhibits early S phase entry observed in Cdh1-depleted cells. The N-terminal region of FoxM1 contains both destruction box (D box) and KEN box sequences that are required for targeting by Cdh1. Mutation of either the D box sequence or the KEN box sequence stabilized FoxM1 and blocked Cdh1-induced proteolysis. Cells expressing a nondegradable form of FoxM1 entered S phase rapidly following release from M phase arrest. Together, our observations show that FoxM1 is one of the targets of Cdh1 in late M or early G(1) phase and that its proteolysis is important for regulated entry into S phase.

Development of a rapid and quantitative method for the analysis of viral entry and release using a NanoLuc luciferase complementation assay.

PubMed

Sasaki, Michihito; Anindita, Paulina D; Phongphaew, Wallaya; Carr, Michael; Kobayashi, Shintaro; Orba, Yasuko; Sawa, Hirofumi

2018-01-02

Subviral particles (SVPs) self-assemble and are released from cells transfected with expression plasmids encoding flavivirus structural proteins. Flavivirus-like particles (VLPs), consisting of flavivirus structural proteins and a subgenomic replicon, can enter cells and cause single-round infections. Neither SVPs or VLPs possess complete viral RNA genomes, therefore are replication-incompetent systems; however, they retain the capacity to fuse and bud from target cells and follow the same maturation process as whole virions. SVPs and VLPs have been previously employed in studies analyzing entry and release steps of viral life cycles. In this study, we have developed quantitative methods for the detection of cellular entry and release of SVPs and VLPs by applying a luciferase complementation assay based on the high affinity interaction between the split NanoLuc luciferase protein, LgBiT and the small peptide, HiBiT. We introduced HiBiT into the structural protein of West Nile virus and generated SVPs and VLPs harboring HiBiT (SVP-HiBiT and VLP-HiBiT, respectively). As SVP-HiBiT emitted strong luminescence upon exposure to LgBiT and its substrate, the nascently budded SVP-HiBiT in the supernatant was readily quantified by luminometry. Similarly, the cellular entry of VLP-HiBiT generated luminescence when VLP-HiBiT was infected into LgBiT-expressing cells. These methods utilizing SVP-HiBiT and VLP-HiBiT will facilitate research into life cycles of flaviviruses, including WNV. Copyright © 2017 Elsevier B.V. All rights reserved.

50 CFR 300.189 - Prohibitions.

Code of Federal Regulations, 2014 CFR

2014-10-01

... under § 300.182. (b) Import as an entry for consumption, purchase, receive for export, export, or re... are imported, entered for consumption, exported, or re-exported. (g) Fail to comply with the documentation requirements as specified in § 300.186, for the importation, entry for consumption, exportation...

50 CFR 300.189 - Prohibitions.

Code of Federal Regulations, 2012 CFR

2012-10-01

... under § 300.182. (b) Import as an entry for consumption, purchase, receive for export, export, or re... are imported, entered for consumption, exported, or re-exported. (g) Fail to comply with the documentation requirements as specified in § 300.186, for the importation, entry for consumption, exportation...

50 CFR 300.189 - Prohibitions.

Code of Federal Regulations, 2013 CFR

2013-10-01

... under § 300.182. (b) Import as an entry for consumption, purchase, receive for export, export, or re... are imported, entered for consumption, exported, or re-exported. (g) Fail to comply with the documentation requirements as specified in § 300.186, for the importation, entry for consumption, exportation...

Putting principals back into practice: an evaluation of a re-entry course for vocationally trained doctors.

PubMed Central

Baker, M; Williams, J; Petchey, R

1997-01-01

BACKGROUND: Current recruitment difficulties in general practice have sharpened the interest of the profession in non-principals. No re-entry course for general practice has previously been run in the UK. AIM: To design and evaluate a re-entry course for general practice. METHOD: A re-entry course was developed to help doctors return to general practice as principals. A telephone interview was carried out with each delegate prior to their attendance on the course and was repeated one month and six months after the course to measure any change in career intentions and the perceived benefit of attending the course. RESULTS: Six months after the course, 11 out of 14 delegates had taken positive steps to return to general practice or had increased their time commitment to medicine. This contrasts with only one of the control group having made any steps to change career. CONCLUSION: The course was evaluated and found to be beneficial, particularly in terms of increasing the confidence of the delegates. PMID:9463984

DOE Office of Scientific and Technical Information (OSTI.GOV)

Airoldi, Flavio, E-mail: flavio.airoldi@multimedica.it; Faglia, Ezio, E-mail: ezio.faglia@multimedica.it; Losa, Sergio, E-mail: sergio.losa@multimedica.it

Subintimal angioplasty (SAP) is frequently performed for the treatment of critical limb ischemia (CLI) and has been recognized as an effective technique for these patients. Nevertheless, this approach is limited by the lack of controlled re-entry into the true lumen of the target vessel. We describe a novel device for true lumen re-entry after subintimal recanalization of superficial femoral arteries (SFA). We report our experience with six patients treated between April 2009 and January 2010 with a novel system designed to facilitate true lumen re-entry. The device was advanced by ipsilateral antegrade approach through a 6-French sheath. Successful reaccess intomore » the true lumen was obtained in five of six patients without complications. The patient in whom the reaccess to the true lumen was not possible underwent successful bypass surgery. At 30 days follow-up, the SFA was patent in all patients according to echo-Doppler examination. Our preliminary experience indicates that this novel re-entry device increases the success rate of percutaneous revascularization of chronically occluded SFA.« less

Thermal Vacuum Facility for Testing Thermal Protection Systems

NASA Technical Reports Server (NTRS)

Daryabeigi, Kamran; Knutson, Jeffrey R.; Sikora, Joseph G.

2002-01-01

A thermal vacuum facility for testing launch vehicle thermal protection systems by subjecting them to transient thermal conditions simulating re-entry aerodynamic heating is described. Re-entry heating is simulated by controlling the test specimen surface temperature and the environmental pressure in the chamber. Design requirements for simulating re-entry conditions are briefly described. A description of the thermal vacuum facility, the quartz lamp array and the control system is provided. The facility was evaluated by subjecting an 18 by 36 in. Inconel honeycomb panel to a typical re-entry pressure and surface temperature profile. For most of the test duration, the average difference between the measured and desired pressures was 1.6% of reading with a standard deviation of +/- 7.4%, while the average difference between measured and desired temperatures was 7.6% of reading with a standard deviation of +/- 6.5%. The temperature non-uniformity across the panel was 12% during the initial heating phase (t less than 500 sec.), and less than 2% during the remainder of the test.

Cell Sage Re Mix HD

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gering, Kevin

Cell Sage is a computational tool that allows science-based evaluation and exploration of battery life cycles over near limitless combination of aging conditions which helps battery end-users to optimize their investments.

Assessment of DNA replication in central nervous system by Laser Scanning Cytometry

NASA Astrophysics Data System (ADS)

Lenz, Dominik; Mosch, Birgit; Bocsi, Jozsef; Arendt, Thomas; Tárnok, Attila

2004-07-01

μIn neurons of patients with Alzheimers's disease (AD) signs of cell cycle re-entry as well as polyploidy have been reported1, 2, indicating that the entire or a part of the genome of the neurons is duplicated before its death but mitosis is not initiated so that the cellular DNA content remains tetraploid. It was concluded, that this imbalance is the direct cause of the neuronal loss in AD3. Manual counting of polyploidal cells is possible but time consuming and possibly statistically insufficient. The aim of this study was to develop an automated method that detects the neuronal DNA content abnormalities with Laser Scanning Cytometry (LSC).Frozen sections of formalin-fixed brain tissue of AD patients and control subjects were labelled with anti-cyclin B and anti-NeuN antibodies. Immunolabelling was performed using Cy5- and Cy2-conjugated secondary antibodies and biotin streptavidin or tyramid signal amplification. In the end sections of 20m thickness were incubated with propidium iodide (PI) (50μg/ml) and covered on slides. For analysis by the LSC PI was used as trigger. Cells identified as neurons by NeuN expression were analyzed for cyclin B expression. Per specimen data of at least 10,000 neurons were acquired. In the frozen brain sections an automated quantification of the amount of nuclear DNA is possible with LSC. The DNA ploidy as well as the cell cycle distribution can be analyzed. A high number of neurons can be scanned and the duration of measuring is shorter than a manual examination. The amount of DNA is sufficiently represented by the PI fluorescence to be able to distinguish between eu- and polyploid neurons.

Returning nurses to the workforce: developing a fast track back program.

PubMed

Burns, Helen K; Sakraida, Teresa J; Englert, Nadine C; Hoffmann, Rosemary L; Tuite, Patricia; Foley, Susan M

2006-01-01

Fast Track Back: Re-entry into Nursing Practice program. Describes the development, implementation, and evaluation of a state-of-the-art re-entry program facilitating the return of licensed nonpracticing RNs to the workforce through a quality education program that retools them for the workforce in the areas of pharmacology, skill development using the latest technology, practice standards, and nursing issues. The program consists of didactic content taught via classroom, Internet, skills laboratory, and high fidelity human simulated technology and a clinical component. The program is a mechanism that enables re-entry nurses to improve skills and competencies necessary to practice in today's healthcare environment.

PLK1 Activation in Late G2 Sets Up Commitment to Mitosis.

PubMed

Gheghiani, Lilia; Loew, Damarys; Lombard, Bérangère; Mansfeld, Jörg; Gavet, Olivier

2017-06-06

Commitment to mitosis must be tightly coordinated with DNA replication to preserve genome integrity. While we have previously established that the timely activation of CyclinB1-Cdk1 in late G2 triggers mitotic entry, the upstream regulatory mechanisms remain unclear. Here, we report that Polo-like kinase 1 (Plk1) is required for entry into mitosis during an unperturbed cell cycle and is rapidly activated shortly before CyclinB1-Cdk1. We determine that Plk1 associates with the Cdc25C1 phosphatase and induces its phosphorylation before mitotic entry. Plk1-dependent Cdc25C1 phosphosites are sufficient to promote mitotic entry, even when Plk1 activity is inhibited. Furthermore, we find that activation of Plk1 during G2 relies on CyclinA2-Cdk activity levels. Our findings thus elucidate a critical role for Plk1 in CyclinB1-Cdk1 activation and mitotic entry and outline how CyclinA2-Cdk, an S-promoting factor, poises cells for commitment to mitosis. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Stochastic E2F activation and reconciliation of phenomenological cell-cycle models.

PubMed

Lee, Tae J; Yao, Guang; Bennett, Dorothy C; Nevins, Joseph R; You, Lingchong

2010-09-21

The transition of the mammalian cell from quiescence to proliferation is a highly variable process. Over the last four decades, two lines of apparently contradictory, phenomenological models have been proposed to account for such temporal variability. These include various forms of the transition probability (TP) model and the growth control (GC) model, which lack mechanistic details. The GC model was further proposed as an alternative explanation for the concept of the restriction point, which we recently demonstrated as being controlled by a bistable Rb-E2F switch. Here, through a combination of modeling and experiments, we show that these different lines of models in essence reflect different aspects of stochastic dynamics in cell cycle entry. In particular, we show that the variable activation of E2F can be described by stochastic activation of the bistable Rb-E2F switch, which in turn may account for the temporal variability in cell cycle entry. Moreover, we show that temporal dynamics of E2F activation can be recast into the frameworks of both the TP model and the GC model via parameter mapping. This mapping suggests that the two lines of phenomenological models can be reconciled through the stochastic dynamics of the Rb-E2F switch. It also suggests a potential utility of the TP or GC models in defining concise, quantitative phenotypes of cell physiology. This may have implications in classifying cell types or states.

Mediator can regulate mitotic entry and direct periodic transcription in fission yeast.

PubMed

Banyai, Gabor; Lopez, Marcela Davila; Szilagyi, Zsolt; Gustafsson, Claes M

2014-11-01

Cdk8 is required for correct timing of mitotic progression in fission yeast. How the activity of Cdk8 is regulated is unclear, since the kinase is not activated by T-loop phosphorylation and its partner, CycC, does not oscillate. Cdk8 is, however, a component of the multiprotein Mediator complex, a conserved coregulator of eukaryotic transcription that is connected to a number of intracellular signaling pathways. We demonstrate here that other Mediator components regulate the activity of Cdk8 in vivo and thereby direct the timing of mitotic entry. Deletion of Mediator components Med12 and Med13 leads to higher cellular Cdk8 protein levels, premature phosphorylation of the Cdk8 target Fkh2, and earlier entry into mitosis. We also demonstrate that Mediator is recruited to clusters of mitotic genes in a periodic fashion and that the complex is required for the transcription of these genes. We suggest that Mediator functions as a hub for coordinated regulation of mitotic progression and cell cycle-dependent transcription. The many signaling pathways and activator proteins shown to function via Mediator may influence the timing of these cell cycle events. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Targeting CCl4 -induced liver fibrosis by RNA interference-mediated inhibition of cyclin E1 in mice.

PubMed

Bangen, Jörg-Martin; Hammerich, Linda; Sonntag, Roland; Baues, Maike; Haas, Ute; Lambertz, Daniela; Longerich, Thomas; Lammers, Twan; Tacke, Frank; Trautwein, Christian; Liedtke, Christian

2017-10-01

Initiation and progression of liver fibrosis requires proliferation and activation of resting hepatic stellate cells (HSCs). Cyclin E1 (CcnE1) is the regulatory subunit of the cyclin-dependent kinase 2 (Cdk2) and controls cell cycle re-entry. We have recently shown that genetic inactivation of CcnE1 prevents activation, proliferation, and survival of HSCs and protects from liver fibrogenesis. The aim of the present study was to translate these findings into preclinical applications using an RNA interference (RNAi)-based approach. CcnE1-siRNA (small interfering RNA) efficiently inhibited CcnE1 gene expression in murine and human HSC cell lines and in primary HSCs, resulting in diminished proliferation and increased cell death. In C57BL/6 wild-type (WT) mice, delivery of stabilized siRNA using a liposome-based carrier targeted approximately 95% of HSCs, 70% of hepatocytes, and 40% of CD45 + cells after single injection. Acute CCl 4 -mediated liver injury in WT mice induced endogenous CcnE1 expression and proliferation of surviving hepatocytes and nonparenchymal cells, including CD45 + leukocytes. Pretreatment with CcnE1-siRNA reverted CcnE1 induction to baseline levels of healthy mice, which was associated with reduced liver injury, diminished proliferation of hepatocytes and leukocytes, and attenuated overall inflammatory response. For induction of liver fibrosis, WT mice were challenged with CCl 4 for 4-6 weeks. Co-treatment with CcnE1-siRNA once a week was sufficient to continuously block CcnE1 expression and cell-cycle activity of hepatocytes and nonparenchymal cells, resulting in significantly ameliorated liver fibrosis and inflammation. Importantly, CcnE1-siRNA also prevented progression of liver fibrosis if applied after onset of chronic liver injury. Therapeutic targeting of CcnE1 in vivo using RNAi is feasible and has high antifibrotic activity. (Hepatology 2017;66:1242-1257). © 2017 by the American Association for the Study of Liver Diseases.

7 CFR 1530.110 - Records, certification, and documentation.

Code of Federal Regulations, 2010 CFR

2010-01-01

... SERVICE, DEPARTMENT OF AGRICULTURE THE REFINED SUGAR RE-EXPORT PROGRAM, THE SUGAR CONTAINING PRODUCTS RE... documents to substantiate entries, transfers, exports, or use as appropriate. The Licensing Authority shall...; (3) Include all U.S. Customs forms submitted in the entry or export process; (4) Provide the correct...

7 CFR 1530.110 - Records, certification, and documentation.

Code of Federal Regulations, 2011 CFR

2011-01-01

... SERVICE, DEPARTMENT OF AGRICULTURE THE REFINED SUGAR RE-EXPORT PROGRAM, THE SUGAR CONTAINING PRODUCTS RE... documents to substantiate entries, transfers, exports, or use as appropriate. The Licensing Authority shall...; (3) Include all U.S. Customs forms submitted in the entry or export process; (4) Provide the correct...

7 CFR 1530.110 - Records, certification, and documentation.

Code of Federal Regulations, 2013 CFR

2013-01-01

... SERVICE, DEPARTMENT OF AGRICULTURE THE REFINED SUGAR RE-EXPORT PROGRAM, THE SUGAR CONTAINING PRODUCTS RE... documents to substantiate entries, transfers, exports, or use as appropriate. The Licensing Authority shall...; (3) Include all U.S. Customs forms submitted in the entry or export process; (4) Provide the correct...

7 CFR 1530.109 - Reporting.

Code of Federal Regulations, 2012 CFR

2012-01-01

... AGRICULTURE THE REFINED SUGAR RE-EXPORT PROGRAM, THE SUGAR CONTAINING PRODUCTS RE-EXPORT PROGRAM, AND THE... transaction; (2) The date of the entry, transfer (only a refiner shall report transfers to the Licensing... license number; (5) The country of origin (entry of raw sugar) or final destination (refined exports...

7 CFR 1530.110 - Records, certification, and documentation.

Code of Federal Regulations, 2014 CFR

2014-01-01

... SERVICE, DEPARTMENT OF AGRICULTURE THE REFINED SUGAR RE-EXPORT PROGRAM, THE SUGAR CONTAINING PRODUCTS RE... documents to substantiate entries, transfers, exports, or use as appropriate. The Licensing Authority shall...; (3) Include all U.S. Customs forms submitted in the entry or export process; (4) Provide the correct...

7 CFR 1530.109 - Reporting.

Code of Federal Regulations, 2013 CFR

2013-01-01

... AGRICULTURE THE REFINED SUGAR RE-EXPORT PROGRAM, THE SUGAR CONTAINING PRODUCTS RE-EXPORT PROGRAM, AND THE... transaction; (2) The date of the entry, transfer (only a refiner shall report transfers to the Licensing... license number; (5) The country of origin (entry of raw sugar) or final destination (refined exports...

7 CFR 1530.109 - Reporting.

Code of Federal Regulations, 2014 CFR

2014-01-01

... AGRICULTURE THE REFINED SUGAR RE-EXPORT PROGRAM, THE SUGAR CONTAINING PRODUCTS RE-EXPORT PROGRAM, AND THE... transaction; (2) The date of the entry, transfer (only a refiner shall report transfers to the Licensing... license number; (5) The country of origin (entry of raw sugar) or final destination (refined exports...

7 CFR 1530.110 - Records, certification, and documentation.

Code of Federal Regulations, 2012 CFR

2012-01-01

... SERVICE, DEPARTMENT OF AGRICULTURE THE REFINED SUGAR RE-EXPORT PROGRAM, THE SUGAR CONTAINING PRODUCTS RE... documents to substantiate entries, transfers, exports, or use as appropriate. The Licensing Authority shall...; (3) Include all U.S. Customs forms submitted in the entry or export process; (4) Provide the correct...

7 CFR 1530.109 - Reporting.

Code of Federal Regulations, 2011 CFR

2011-01-01

... AGRICULTURE THE REFINED SUGAR RE-EXPORT PROGRAM, THE SUGAR CONTAINING PRODUCTS RE-EXPORT PROGRAM, AND THE... transaction; (2) The date of the entry, transfer (only a refiner shall report transfers to the Licensing... license number; (5) The country of origin (entry of raw sugar) or final destination (refined exports...

7 CFR 1530.109 - Reporting.

Code of Federal Regulations, 2010 CFR

2010-01-01

... AGRICULTURE THE REFINED SUGAR RE-EXPORT PROGRAM, THE SUGAR CONTAINING PRODUCTS RE-EXPORT PROGRAM, AND THE... transaction; (2) The date of the entry, transfer (only a refiner shall report transfers to the Licensing... license number; (5) The country of origin (entry of raw sugar) or final destination (refined exports...

Japanese encephalitis virus invasion of cell: allies and alleys.

PubMed

Nain, Minu; Abdin, Malik Z; Kalia, Manjula; Vrati, Sudhanshu

2016-03-01

The mosquito-borne flavivirus, Japanese encephalitis virus (JEV), is the leading cause of virus-induced encephalitis globally and a major public health concern of several countries in Southeast Asia, with the potential to become a global pathogen. The virus is neurotropic, and the disease ranges from mild fever to severe hemorrhagic and encephalitic manifestations and death. The early steps of the virus life cycle, binding, and entry into the cell are crucial determinants of infection and are potential targets for the development of antiviral therapies. JEV can infect multiple cell types; however, the key receptor molecule(s) still remains elusive. JEV also has the capacity to utilize multiple endocytic pathways for entry into cells of different lineages. This review not only gives a comprehensive update on what is known about the virus attachment and receptor system (allies) and the endocytic pathways (alleys) exploited by the virus to gain entry into the cell and establish infection but also discusses crucial unresolved issues. We also highlight common themes and key differences between JEV and other flaviviruses in these contexts. Copyright © 2015 John Wiley & Sons, Ltd.

Inhibitory effects of (-)-epigallocatechin gallate on the life cycle of human immunodeficiency virus type 1 (HIV-1).

PubMed

Yamaguchi, Koushi; Honda, Mitsuo; Ikigai, Hajime; Hara, Yukihiko; Shimamura, Tadakatsu

2002-01-01

Epigallocatechin gallate (EGCg), the major tea catechin, is known as a potent anti-bacterial agent. In addition, anti-tumor promoting, anti-inflammatory, anti-oxidative and antiviral activities have been reported. In the present study, we investigated possible anti-human immunodeficiency virus type-1 (HIV-1) activity of EGCg and its mechanisms of action in the viral life cycle. EGCg impinges on each step of the HIV life cycle. Thus, destruction of the viral particles, viral attachment to cells, post-adsorption entry into cells, reverse transcription (RT), viral production from chronically-infected cells, and the level of expression of viral mRNA, were analyzed using T-lymphoid (H9) and monocytoid (THP-1) cell systems, and antiviral protease activity was measured using a cell-free assay. Inhibitory effects of EGCg on specific binding of the virions to the cellular surfaces and changes in the steady state viral regulation (mRNA expression) due to EGCg were not observed. However, EGCg had a destructive effect on the viral particles, and post-adsorption entry and RT in acutely infected monocytoid cells were significantly inhibited at concentrations of EGCg greater than 1 microM, and protease kinetics were suppressed at a concentration higher than 10 microM in the cell-free study. Viral production by THP-1 cells chronically-infected with HIV-1 was also inhibited in a dose-dependent manner and the inhibitory effect was enhanced by liposome modification of EGCg. As expected, increased viral mRNA production was observed in lipopolysaccharide (LPS)-activated chronically HIV-1-infected cells. This production was significantly inhibited by EGCg treatment of THP-1 cells. In contrast, production of HIV-1 viral mRNA in unstimulated or LPS-stimulated T-lymphoid cells (H9) was not inhibited by EGCg. Anti-HIV viral activity of EGCg may thus result from an interaction with several steps in the HIV-1 life cycle.

On the entry of an emerging arbovirus into host cells: Mayaro virus takes the highway to the cytoplasm through fusion with early endosomes and caveolae-derived vesicles

PubMed Central

Carvalho, Carlos A.M.; Silva, Jerson L.; Oliveira, Andréa C.

2017-01-01

Mayaro virus (MAYV) is an emergent sylvatic alphavirus in South America, related to sporadic outbreaks of a chikungunya-like human febrile illness accompanied by severe arthralgia. Despite its high potential for urban emergence, MAYV is still an obscure virus with scarce information about its infection cycle, including the corresponding early events. Even for prototypical alphaviruses, the cell entry mechanism still has some rough edges to trim: although clathrin-mediated endocytosis is quoted as the putative route, alternative paths as distinct as direct virus genome injection through the cell plasma membrane seems to be possible. Our aim was to clarify crucial details on the entry route exploited by MAYV to gain access into the host cell. Tracking the virus since its first contact with the surface of Vero cells by fluorescence microscopy, we show that its entry occurs by a fast endocytic process and relies on fusion with acidic endosomal compartments. Moreover, blocking clathrin-mediated endocytosis or depleting cholesterol from the cell membrane leads to a strong inhibition of viral infection, as assessed by plaque assays. Following this clue, we found that early endosomes and caveolae-derived vesicles are both implicated as target membranes for MAYV fusion. Our findings unravel the very first events that culminate in a productive infection by MAYV and shed light on potential targets for a rational antiviral therapy, besides providing a better comprehension of the entry routes exploited by alphaviruses to get into the cell. PMID:28462045

Seismic Parameters of Mining-Induced Aftershock Sequences for Re-entry Protocol Development

NASA Astrophysics Data System (ADS)

Vallejos, Javier A.; Estay, Rodrigo A.

2018-03-01

A common characteristic of deep mines in hard rock is induced seismicity. This results from stress changes and rock failure around mining excavations. Following large seismic events, there is an increase in the levels of seismicity, which gradually decay with time. Restricting access to areas of a mine for enough time to allow this decay of seismic events is the main approach in re-entry strategies. The statistical properties of aftershock sequences can be studied with three scaling relations: (1) Gutenberg-Richter frequency magnitude, (2) the modified Omori's law (MOL) for the temporal decay, and (3) Båth's law for the magnitude of the largest aftershock. In this paper, these three scaling relations, in addition to the stochastic Reasenberg-Jones model are applied to study the characteristic parameters of 11 large magnitude mining-induced aftershock sequences in four mines in Ontario, Canada. To provide guidelines for re-entry protocol development, the dependence of the scaling relation parameters on the magnitude of the main event are studied. Some relations between the parameters and the magnitude of the main event are found. Using these relationships and the scaling relations, a space-time-magnitude re-entry protocol is developed. These findings provide a first approximation to concise and well-justified guidelines for re-entry protocol development applicable to the range of mining conditions found in Ontario, Canada.

Problem-based test: replication of mitochondrial DNA during the cell cycle.

PubMed

Sétáló, György

2013-01-01

Terms to be familiar with before you start to solve the test: cell cycle, generation time, S-phase, cell culture synchronization, isotopic pulse-chase labeling, density labeling, equilibrium density-gradient centrifugation, buoyant density, rate-zonal centrifugation, nucleoside, nucleotide, kinase enzymes, polymerization of nucleic acids, re-replication block, cell fractionation, Svedberg (sedimentation constant = [ S]), nuclear DNA, mitochondrial DNA, heavy and light mitochondrial DNA chains, heteroplasmy, mitochondrial diseases Copyright © 2013 Wiley Periodicals, Inc.

A Polarized Cell Model for Chikungunya Virus Infection: Entry and Egress of Virus Occurs at the Apical Domain of Polarized Cells

PubMed Central

Lim, Pei Jin; Chu, Justin Jang Hann

2014-01-01

Chikungunya virus (CHIKV) has resulted in several outbreaks in the past six decades. The clinical symptoms of Chikungunya infection include fever, skin rash, arthralgia, and an increasing incidence of encephalitis. The re-emergence of CHIKV with more severe pathogenesis highlights its potential threat on our human health. In this study, polarized HBMEC, polarized Vero C1008 and non-polarized Vero cells grown on cell culture inserts were infected with CHIKV apically or basolaterally. Plaque assays, viral binding assays and immunofluorescence assays demonstrated apical entry and release of CHIKV in polarized HBMEC and Vero C1008. Drug treatment studies were performed to elucidate both host cell and viral factors involved in the sorting and release of CHIKV at the apical domain of polarized cells. Disruption of host cell myosin II, microtubule and microfilament networks did not disrupt the polarized release of CHIKV. However, treatment with tunicamycin resulted in a bi-directional release of CHIKV, suggesting that N-glycans of CHIKV envelope glycoproteins could serve as apical sorting signals. PMID:24587455

Characterization of the "CCR5" Chemokine Receptor Gene

ERIC Educational Resources Information Center

Thomas, John C.

2004-01-01

The life cycle of retroviruses is an essential topic of modern cell biology instruction. Furthermore, the process of HIV viral entry into the cell is a question of great interest in basic and clinical biology. This paper describes how students can easily recover their own DNA, amplify a portion of the "CCR5" chemokine receptor gene, characterize…

Amiodarone affects Ebola virus binding and entry into target cells.

PubMed

Salata, Cristiano; Munegato, Denis; Martelli, Francesco; Parolin, Cristina; Calistri, Arianna; Baritussio, Aldo; Palù, Giorgio

2018-03-02

Ebola Virus Disease is one of the most lethal transmissible infections characterized by a high fatality rate. Several research studies have aimed to identify effective antiviral agents. Amiodarone, a drug used for the treatment of arrhythmias, has been shown to inhibit filovirus infection in vitro by acting at the early step of the viral replication cycle. Here we demonstrate that amiodarone reduces virus binding to target cells and slows down the progression of the viral particles along the endocytic pathway. Overall our data support the notion that amiodarone interferes with Ebola virus infection by affecting cellular pathways/targets involved in the viral entry process.

Dynamic and Static High Temperature Resistant Ceramic Seals for X- 38 re-Entry Vehicle

NASA Astrophysics Data System (ADS)

Handrick, Karin E.; Curry, Donald M.

2002-01-01

In a highly successful partnership, NAS A, ESA, DLR (German Space Agency) and European industry are building the X-38, V201 re-entry spacecraft, the prototype of the International Space Station's Crew Return Vehicle (CRV). This vehicle would serve both as an ambulance for medical emergencies and as an evacuation vehicle for the Space Station. The development of essential systems and technologies for a reusable re-entry vehicle is a first for Europe, and sharing the development of an advanced re-entry spacecraft with foreign partners is a first for NASA. NASA, in addition to its subsystem responsibilities, is performing overall X-38 vehicle system engineering and integration, will launch V201 on the Space Shuttle, deliver flight data for post-flight analysis and assessment and is responsible for development and manufacture of structural vehicle components and thermal protection (TPS) tiles. The major European objective for cooperation with NASA on X-38 was to establish a clear path through which key technologies needed for future space transportation systems could be developed and validated at affordable cost and with controlled risk. Europe has taken the responsibility to design and manufacture hot control surfaces like metallic rudders and ceramic matrix composites (CMC) body flaps, thermal protection systems such as CMC leading edges, the CMC nose cap and -skirt, insulation, landing gears and elements of the V201 primary structure. Especially hot control surfaces require extremely high temperature resistant seals to limit hot gas ingestion and transfer of heat to underlying low-temperature structures to prevent overheating of these structures and possible loss of the vehicle. Complex seal interfaces, which have to fulfill various, tight mission- and vehicle-related requirements exist between the moveable ceramic body flaps and the bottom surface of the vehicle, between the rudder and fin structure and the ceramic leading edge panel and TPS tiles. While NASA concentrated on the development, qualification and manufacture of dynamic seals in the rudder area, the responsibility of MAN Technologie focused on the development, lay-out, qualification and flight hardware manufacture of static and dynamic seals in ceramic hot structures' associated gaps and interfaces, dealing with re-entry temperatures up to 1600°C. This paper presents results for temperature and mechanical stability, flow, scrub (up to 1000 cycles) and of arc jet tests under representative low boundary conditions and plasma step/gap tests, conducted during the development and qualification phases of these different kind of ceramic seals. Room temperature seal compression tests were performed at low compression levels to determine load versus linear compression, preload, contact area, stiffness and resiliency characteristics under low load conditions. Flow tests with thermally aged seals were conducted at ambient temperature to examine leakage at low compression levels and in as-manufactured conditions. Seal scrub tests were performed to examine durability and wear resistance and to recommend surface treatments required to maximize seal wear life. Results of arc jet/plasma tests under simulated re-entry conditions (pressure, temperature) verified seal temperature stability and function under representative assembly and interface conditions. Each of these specifically developed seals fulfilled the requirements and is qualified for flight on X-38, V201.

Hormonal Involvement in Breast Cancer Gene Amplification

DTIC Science & Technology

2008-10-01

and transcriptome. Genome Res.16: 394-404. Zhu W. and Dutta A. (2006). Activation of Fanconi anemia pathway in cells with re- replicated DNA. Cell Cycle 5: 2306-2309. APPENDICES None SUPPORTING DATA None

Quantitative membrane proteomics reveals a role for tetraspanin enriched microdomains during entry of human cytomegalovirus

PubMed Central

John, Nessy; Malouli, Daniel

2017-01-01

Human cytomegalovirus (HCMV) depends on and modulates multiple host cell membrane proteins during each stage of the viral life cycle. To gain a global view of the impact of HCMV-infection on membrane proteins, we analyzed HCMV-induced changes in the abundance of membrane proteins in fibroblasts using stable isotope labeling with amino acids (SILAC), membrane fractionation and protein identification by two-dimensional liquid chromatography and tandem mass spectrometry. This systematic approach revealed that CD81, CD44, CD98, caveolin-1 and catenin delta-1 were down-regulated during infection whereas GRP-78 was up-regulated. Since CD81 downregulation was also observed during infection with UV-inactivated virus we hypothesized that this tetraspanin is part of the viral entry process. Interestingly, additional members of the tetraspanin family, CD9 and CD151, were also downregulated during HCMV-entry. Since tetraspanin-enriched microdomains (TEM) cluster host cell membrane proteins including known CMV receptors such as integrins, we studied whether TEMs are required for viral entry. When TEMs were disrupted with the cholesterol chelator methyl-β-cylcodextrin, viral entry was inhibited and this inhibition correlated with reduced surface levels of CD81, CD9 and CD151, whereas integrin levels remained unchanged. Furthermore, simultaneous siRNA-mediated knockdown of multiple tetraspanins inhibited viral entry whereas individual knockdown had little effect suggesting essential, but redundant roles for individual tetraspanins during entry. Taken together, our data suggest that TEM act as platforms for receptors utilized by HCMV for entry into cells. PMID:29121670

PIRATES: A Program for Offenders Transitioning into the World of Work

ERIC Educational Resources Information Center

Musgrove, Kate Racoff; Derzis, Nicholas C.; Shippen, Margaret E.; Brigman, Holly E.

2012-01-01

The study assessed the effectiveness of the Preparing Inmates for Re-Entry through Assistance, Training, and Employment Skills (PIRATES) group, which focused on improving dysfunctional career thoughts. Participants were male offenders (n = 14) enrolled in a re-entry training class. Results showed a statistically significant decrease in…

Effects of Angle of Attack on the Behaviour of Imperfections in Thermal Protection Systems of Re-entry Vehicles

NASA Astrophysics Data System (ADS)

Palharini, R. C.; Scanlon, T. J.; Reese, J. M.

The study of atmospheric re-entry under rarefied nonequilibrium flows is a challenging problem directly related to the development of new aerospace technologies, where the prediction of thermal loads acting over the spacecraft is critical during descent phase.

Sacramento City College Re-Entry Services Comprehensive Plan.

ERIC Educational Resources Information Center

White, Maureen E.; Smith, William A.

Sacramento City College (SCC) established its Re-Entry Services program to provide information, referral and support services to students returning to the academic environment after an absence. Since the inception of the program in 1977, the college community has changed considerably. Among these changes are an aging student population, increased…

Study and Development of a Sub-Orbital Re-Entry Demonstrator

NASA Astrophysics Data System (ADS)

Savino, R.

The Italian and European Space Agencies are supporting a research programme, developed in Campania region by a cluster of industries, research institutes and universities, on a low-cost re-entry capsule, able to return payloads from the ISS to Earth and/or to perform short-duration scientific missions in Low Earth Orbit (LEO). The ballistic capsule is characterized by a deployable, disposable "umbrella-like" heat shield that allows relatively small dimensions at launch and a sufficient exposed surface area in re-entry conditions, reducing the ballistic coefficient and leading to acceptable heat fluxes, mechanical loads and final descent velocity. ESA is supporting a preliminary study to develop a flight demonstrator of the capsule to be embarked as a secondary payload onboard a sub-orbital sounding rocket. The deployable thermal protection system concept may be applied to future science and robotic exploration mission requiring planetary entry and, possibly also to missions in the framework of Human Space flight, requiring planetary entry or re-entry. The technology offers also an interesting potential for aerobraking, aerocapture and for de-orbiting. This paper summarizes the results of these activities, which are being more and more refined as the work proceeds, including the definition and analysis of the mission scenario, the aerodynamic, aerothermodynamic, mechanical and structural analyses and the technical definition of avionics, instrumentation and main subsystems.

Imaging single cells in a beam of live cyanobacteria with an X-ray laser (CXIDB ID 26)

DOE Data Explorer

Schot, Gijs, vander

2015-02-10

This entry contains ten diffraction patterns, and reconstructions images, of individual living Cyanobium gracile cells, imaged using 517 eV X-rays from the LCLS XFEL. The Hawk software package was used for phasing. The Uppsala aerosol injector was used for sample injection, assuring very low noise levels. The cells come from various stages of the cell cycle, and were imaged in random orientations.

Subintimal angioplasty with the aid of a re-entry device for TASC C and D lesions of the SFA.

PubMed

Setacci, C; Chisci, E; de Donato, G; Setacci, F; Iacoponi, F; Galzerano, G

2009-07-01

The aim of this prospective study was to assess the clinical effectiveness and related midterm patency of subintimal angioplasty (SAP) in patients suffering from critical limb ischaemia (CLI) in a single tertiary care university centre. The secondary aim was to evaluate the safety and clinical effectiveness of using a re-entry device when re-canalisation by SAP was unsuccessful. From January 2005 to December 2007, consecutive patients suffering from CLI (Rutherford clinical categories: 4-6) were treated with SAP. All patients included in the study had occluded SFA (TASC C and D) and underwent clinical and ultrasound follow-up examinations at day 30 and at 3, 6, 9 and 12 months, and then yearly. A re-entry device (Outback, Cordis Corporation, Miami Lakes, Florida, USA in all cases) was only used when re-canalisation by simple SAP was unsuccessful, and stenting was used when residual stenosis was >30% or there was a flow-limiting dissection. Factors that could modify the outcome were analysed. In this study, 145 patients were treated, with a technical success rate of 83.5% (121 of 145) for simple SAP. Stenting was performed in 43% (n=62) of successful SAP procedures. No death occurred in the perioperative period, while the 30-day mortality was 4.8% (7 of 145). The re-entry device (Outback) was used in 24 cases (16.5%). The technical success of the re-entry device was 79% (19 of 24), with a 90% success rate of stent placement at the site of re-entry. Complications occurred in 6.2% of all procedures (n=9) (three arterial perforations (2.1%), three distal embolisations (2.1%), two femoral artery pseudo-aneurysms (1.4%) and one arterio-venous fistula (0.7%)). Factors capable of independently affecting the patency were renal insufficiency (p=0.03), current smoking (p=0.01) and diabetes (p=0.04). The primary patency at 1 and 3 years was 70% and 34% and the secondary patency at 1 and 3 years was 77% and 43%, respectively. At the same time intervals, the limb-salvage rate was 88% and 49%. SAP with the aid of a re-entry device for TASC C and D lesions of the SFA seems to be safe and clinically effective in patients suffering from CLI, according to the experience at our centre. Further follow-up and more data are necessary to confirm these findings.

Endurance Enhancement and High Speed Set/Reset of 50 nm Generation HfO2 Based Resistive Random Access Memory Cell by Intelligent Set/Reset Pulse Shape Optimization and Verify Scheme

NASA Astrophysics Data System (ADS)

Higuchi, Kazuhide; Miyaji, Kousuke; Johguchi, Koh; Takeuchi, Ken

2012-02-01

This paper proposes a verify-programming method for the resistive random access memory (ReRAM) cell which achieves a 50-times higher endurance and a fast set and reset compared with the conventional method. The proposed verify-programming method uses the incremental pulse width with turnback (IPWWT) for the reset and the incremental voltage with turnback (IVWT) for the set. With the combination of IPWWT reset and IVWT set, the endurance-cycle increases from 48 ×103 to 2444 ×103 cycles. Furthermore, the measured data retention-time after 20 ×103 set/reset cycles is estimated to be 10 years. Additionally, the filamentary based physical model is proposed to explain the set/reset failure mechanism with various set/reset pulse shapes. The reset pulse width and set voltage correspond to the width and length of the conductive-filament, respectively. Consequently, since the proposed IPWWT and IVWT recover set and reset failures of ReRAM cells, the endurance-cycles are improved.

A Second Chance to Dream: Initiating ODeL in Secondary School Re-Entry Programs for Young Adult Secondary School Dropouts the Case of Mumias District, Western Kenya

ERIC Educational Resources Information Center

Musita, Richard; Ogange, Betty O.; Lugendo, Dorine

2018-01-01

The Kenyan education system has very limited re-entry options for learners who drop out before attaining secondary school certificate. It is very difficult to access training and or secure a job that requires at least secondary school education. This study examined the prospects of initiating Open and Distance e-Learning(ODeL) in re-entry…

Workforce re-entry for Japanese unemployed dental hygienists.

PubMed

Usui, Y; Miura, H

2015-02-01

The aim of this study was to define the profile of unemployed dental hygienists who could be enticed to re-enter the workforce and the factors that could facilitate their re-entry into the dental field in Japan. The questionnaire was mailed with a postage-paid return envelope to a sample of 3095 licensed dental hygienists. A 50.4% response rate (S = 1477) was observed. The rate of working dental hygienists was 60.3% (n = 891), and of unemployed dental hygienists was 39.7% (n = 586). Of the latter, 31.9% (n = 187) stated intentions of returning to the workplace. The unemployed dental hygienists seeking employment were more often married and had more children, compared with working dental hygienists currently. This group also had significantly fewer total service years. Moreover, only 11.96% of them belonged to the Japan Dental Hygienists' Association, and 41.3% of those attended training workshops. According to their response, they perceived their top three major barriers to re-entry as 'lack sufficient dental hygiene skill', 'child rearing' and 'poor working atmosphere'. 'Flexibility in the work schedule' and 'location' were the most important factors for re-entry from their perspective. There were not many dental hygienists hoping to return to the dental field. The findings suggested that strategies to encourage non-practicing dental hygienists to re-entry should be emphasized in the areas of a flexible working atmosphere, easy access to information on how to return to practice and guidance on how to maintain professionalism during inactivity. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Infection of Mouse Macrophages by Seasonal Influenza Viruses Can Be Restricted at the Level of Virus Entry and at a Late Stage in the Virus Life Cycle

PubMed Central

Londrigan, Sarah L.; Short, Kirsty R.; Ma, Joel; Gillespie, Leah; Rockman, Steven P.; Brooks, Andrew G.

2015-01-01

ABSTRACT Airway epithelial cells are susceptible to infection with seasonal influenza A viruses (IAV), resulting in productive virus replication and release. Macrophages (MΦ) are also permissive to IAV infection; however, virus replication is abortive. Currently, it is unclear how productive infection of MΦ is impaired or the extent to which seasonal IAV replicate in MΦ. Herein, we compared mouse MΦ and epithelial cells for their ability to support genomic replication and transcription, synthesis of viral proteins, assembly of virions, and release of infectious progeny following exposure to genetically defined IAV. We confirm that seasonal IAV differ in their ability to utilize cell surface receptors for infectious entry and that this represents one level of virus restriction. Following virus entry, we demonstrate synthesis of all eight segments of genomic viral RNA (vRNA) and mRNA, as well as seven distinct IAV proteins, in IAV-infected mouse MΦ. Although newly synthesized hemagglutinin (HA) and neuraminidase (NA) glycoproteins are incorporated into the plasma membrane and expressed at the cell surface, electron microscopy confirmed that virus assembly was defective in IAV-infected MΦ, defining a second level of restriction late in the virus life cycle. IMPORTANCE Seasonal influenza A viruses (IAV) and highly pathogenic avian influenza viruses (HPAI) infect macrophages, but only HPAI replicate productively in these cells. Herein, we demonstrate that impaired virus uptake into macrophages represents one level of restriction limiting infection by seasonal IAV. Following uptake, seasonal IAV do not complete productive replication in macrophages, representing a second level of restriction. Using murine macrophages, we demonstrate that productive infection is blocked late in the virus life cycle, such that virus assembly is defective and newly synthesized virions are not released. These studies represent an important step toward identifying host-encoded factors that block replication of seasonal IAV, but not HPAI, in macrophages. PMID:26423941

Infection of Mouse Macrophages by Seasonal Influenza Viruses Can Be Restricted at the Level of Virus Entry and at a Late Stage in the Virus Life Cycle.

PubMed

Londrigan, Sarah L; Short, Kirsty R; Ma, Joel; Gillespie, Leah; Rockman, Steven P; Brooks, Andrew G; Reading, Patrick C

2015-12-01

Airway epithelial cells are susceptible to infection with seasonal influenza A viruses (IAV), resulting in productive virus replication and release. Macrophages (MΦ) are also permissive to IAV infection; however, virus replication is abortive. Currently, it is unclear how productive infection of MΦ is impaired or the extent to which seasonal IAV replicate in MΦ. Herein, we compared mouse MΦ and epithelial cells for their ability to support genomic replication and transcription, synthesis of viral proteins, assembly of virions, and release of infectious progeny following exposure to genetically defined IAV. We confirm that seasonal IAV differ in their ability to utilize cell surface receptors for infectious entry and that this represents one level of virus restriction. Following virus entry, we demonstrate synthesis of all eight segments of genomic viral RNA (vRNA) and mRNA, as well as seven distinct IAV proteins, in IAV-infected mouse MΦ. Although newly synthesized hemagglutinin (HA) and neuraminidase (NA) glycoproteins are incorporated into the plasma membrane and expressed at the cell surface, electron microscopy confirmed that virus assembly was defective in IAV-infected MΦ, defining a second level of restriction late in the virus life cycle. Seasonal influenza A viruses (IAV) and highly pathogenic avian influenza viruses (HPAI) infect macrophages, but only HPAI replicate productively in these cells. Herein, we demonstrate that impaired virus uptake into macrophages represents one level of restriction limiting infection by seasonal IAV. Following uptake, seasonal IAV do not complete productive replication in macrophages, representing a second level of restriction. Using murine macrophages, we demonstrate that productive infection is blocked late in the virus life cycle, such that virus assembly is defective and newly synthesized virions are not released. These studies represent an important step toward identifying host-encoded factors that block replication of seasonal IAV, but not HPAI, in macrophages. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Proteomic analysis of the response to cell cycle arrests in human myeloid leukemia cells.

PubMed

Ly, Tony; Endo, Aki; Lamond, Angus I

2015-01-02

Previously, we analyzed protein abundance changes across a 'minimally perturbed' cell cycle by using centrifugal elutriation to differentially enrich distinct cell cycle phases in human NB4 cells (Ly et al., 2014). In this study, we compare data from elutriated cells with NB4 cells arrested at comparable phases using serum starvation, hydroxyurea, or RO-3306. While elutriated and arrested cells have similar patterns of DNA content and cyclin expression, a large fraction of the proteome changes detected in arrested cells are found to reflect arrest-specific responses (i.e., starvation, DNA damage, CDK1 inhibition), rather than physiological cell cycle regulation. For example, we show most cells arrested in G2 by CDK1 inhibition express abnormally high levels of replication and origin licensing factors and are likely poised for genome re-replication. The protein data are available in the Encyclopedia of Proteome Dynamics (

Assessment Of The Aerodynamic And Aerothermodynamic Performance Of The USV-3 High-Lift Re-Entry Vehicle

NASA Astrophysics Data System (ADS)

Pezzella, Giuseppe; Richiello, Camillo; Russo, Gennaro

2011-05-01

This paper deals with the aerodynamic and aerothermodynamic trade-off analysis carried out with the aim to design a hypersonic flying test bed (FTB), namely USV3. Such vehicle will have to be launched with a small expendable launcher and shall re-enter the Earth atmosphere allowing to perform several experiments on critical re-entry phenomena. The demonstrator under study is a re-entry space glider characterized by a relatively simple vehicle architecture able to validate hypersonic aerothermodynamic design database and passenger experiments, including thermal shield and hot structures. Then, a summary review of the aerodynamic characteristics of two FTB concepts, compliant with a phase-A design level, has been provided hereinafter. Indeed, several design results, based both on engineering approach and computational fluid dynamics, are reported and discussed in the paper.

Cell-cycle dynamics of chromosomal organisation at single-cell resolution

PubMed Central

Nagano, Takashi; Lubling, Yaniv; Várnai, Csilla; Dudley, Carmel; Leung, Wing; Baran, Yael; Mendelson-Cohen, Netta; Wingett, Steven; Fraser, Peter; Tanay, Amos

2017-01-01

Summary Chromosomes in proliferating metazoan cells undergo dramatic structural metamorphoses every cell cycle, alternating between highly condensed mitotic structures facilitating chromosome segregation, and decondensed interphase structures accommodating transcription, gene silencing and DNA replication. Here we use single-cell Hi-C to study chromosome conformations in thousands of individual cells, and discover a continuum of cis-interaction profiles that finely position individual cells along the cell cycle. We show that chromosomal compartments, topological associated domains (TADs), contact insulation and long-range loops, all defined by bulk Hi-C maps, are governed by distinct cell-cycle dynamics. In particular, DNA replication correlates with build-up of compartments and reduction in TAD insulation, while loops are generally stable from G1 through S and G2. Whole-genome 3D structural models reveal a radial architecture of chromosomal compartments with distinct epigenomic signatures. Our single-cell data thereby allow for re-interpretation of chromosome conformation maps through the prism of the cell cycle. PMID:28682332

Levels of Ycg1 Limit Condensin Function during the Cell Cycle

PubMed Central

Arsenault, Heather E.; Benanti, Jennifer A.

2016-01-01

During mitosis chromosomes are condensed to facilitate their segregation, through a process mediated by the condensin complex. Although several factors that promote maximal condensin activity during mitosis have been identified, the mechanisms that downregulate condensin activity during interphase are largely unknown. Here, we demonstrate that Ycg1, the Cap-G subunit of budding yeast condensin, is cell cycle-regulated with levels peaking in mitosis and decreasing as cells enter G1 phase. This cyclical expression pattern is established by a combination of cell cycle-regulated transcription and constitutive degradation. Interestingly, overexpression of YCG1 and mutations that stabilize Ycg1 each result in delayed cell-cycle entry and an overall proliferation defect. Overexpression of no other condensin subunit impacts the cell cycle, suggesting that Ycg1 is limiting for condensin complex formation. Consistent with this possibility, we find that levels of intact condensin complex are reduced in G1 phase compared to mitosis, and that increased Ycg1 expression leads to increases in both levels of condensin complex and binding to chromatin in G1. Together, these results demonstrate that Ycg1 levels limit condensin function in interphase cells, and suggest that the association of condensin with chromosomes must be reduced following mitosis to enable efficient progression through the cell cycle. PMID:27463097

Thermal investigation of nuclear waste disposal in space

NASA Technical Reports Server (NTRS)

Wilkinson, C. L.

1981-01-01

A thermal analysis has been conducted to determine the allowable size and response of bare and shielded nuclear waste forms in both low earth orbit and at 0.85 astronomical units. Contingency conditions of re-entry with a 45 deg and 60 deg aeroshell are examined as well as re-entry of a spherical shielded waste form. A variety of shielded schemes were examined and the waste form thermal response for each determined. Two optimum configurations were selected. The thermal response of these two shielded waste configurations to indefinite exposure to ground conditions following controlled and uncontrolled re-entry is determined. In all cases the prime criterion is that waste containment must be maintained.

Recent advances in understanding of meiosis initiation and the apomictic pathway in plants.

PubMed

Wang, Chung-Ju R; Tseng, Ching-Chih

2014-01-01

Meiosis, a specialized cell division to produce haploid cells, marks the transition from a sporophytic to a gametophytic generation in the life cycle of plants. In angiosperms, meiosis takes place in sporogenous cells that develop de novo from somatic cells in anthers or ovules. A successful transition from the mitotic cycle to the meiotic program in sporogenous cells is crucial for sexual reproduction. By contrast, when meiosis is bypassed or a mitosis-like division occurs to produce unreduced cells, followed by the development of an embryo sac, clonal seeds can be produced by apomixis, an asexual reproduction pathway found in 400 species of flowering plants. An understanding of the regulation of entry into meiosis and molecular mechanisms of apomictic pathway will provide vital insight into reproduction for plant breeding. Recent findings suggest that AM1/SWI1 may be the key gene for entry into meiosis, and increasing evidence has shown that the apomictic pathway is epigenetically controlled. However, the mechanism for the initiation of meiosis during sexual reproduction or for its omission in the apomictic pathway still remains largely unknown. Here we review the current understanding of meiosis initiation and the apomictic pathway and raised several questions that are awaiting further investigation.

Rab5 and Rab11 Are Required for Clathrin-Dependent Endocytosis of Japanese Encephalitis Virus in BHK-21 Cells.

PubMed

Liu, Chun-Chun; Zhang, Yun-Na; Li, Zhao-Yao; Hou, Jin-Xiu; Zhou, Jing; Kan, Lin; Zhou, Bin; Chen, Pu-Yan

2017-10-01

During infection Japanese encephalitis virus (JEV) generally enters host cells via receptor-mediated clathrin-dependent endocytosis. The trafficking of JEV within endosomes is controlled by Rab GTPases, but which Rab proteins are involved in JEV entry into BHK-21 cells is unknown. In this study, entry and postinternalization of JEV were analyzed using biochemical inhibitors, RNA interference, and dominant negative (DN) mutants. Our data demonstrate that JEV entry into BHK-21 cells depends on clathrin, dynamin, and cholesterol but not on caveolae or macropinocytosis. The effect on JEV infection of dominant negative (DN) mutants of four Rab proteins that regulate endosomal trafficking was examined. Expression of DN Rab5 and DN Rab11, but not DN Rab7 and DN Rab9, significantly inhibited JEV replication. These results were further tested by silencing Rab5 or Rab11 expression before viral infection. Confocal microscopy showed that virus particles colocalized with Rab5 or Rab11 within 15 min after virus entry, suggesting that after internalization JEV moves to early and recycling endosomes before the release of the viral genome. Our findings demonstrate the roles of Rab5 and Rab11 on JEV infection of BHK-21 cells through the endocytic pathway, providing new insights into the life cycle of flaviviruses. IMPORTANCE Although Japanese encephalitis virus (JEV) utilizes different endocytic pathways depending on the cell type being infected, the detailed mechanism of its entry into BHK-21 cells is unknown. Understanding the process of JEV endocytosis and postinternalization will advance our knowledge of JEV infection and pathogenesis as well as provide potential novel drug targets for antiviral intervention. With this objective, we used systematic approaches to dissect this process. The results show that entry of JEV into BHK-21 cells requires a low-pH environment and that the process occurs through dynamin-, actin-, and cholesterol-dependent clathrin-mediated endocytosis that requires Rab5 and Rab11. Our work provides a detailed picture of the entry of JEV into BHK-21 cells and the cellular events that follow. Copyright © 2017 American Society for Microbiology.

Insertion of targeting domains into the envelope glycoprotein of Moloney murine leukemia virus (MoMLV)-based vectors modulates the route of mCAT-1-mediated viral entry.

PubMed

Viejo-Borbolla, A; Pizzato, M; Blair, E D; Schulz, T F

2005-03-01

Several groups have inserted targeting domains into the envelope glycoprotein (Env) of Moloney murine leukemia virus (MoMLV) in an attempt to produce targeted retroviral vectors for human gene therapy. While binding of these modified Envs to the target molecule expressed on the surface of human cells was observed, specific high-titer infection of human cells expressing the target molecule was not achieved. Here we investigate the initial steps in the entry process of targeted MoMLV vectors both in murine and human cells expressing the MoMLV receptor, the mouse cationic amino acid transporter-1 (mCAT-1). We show that insertion of a small ligand targeted to E-selectin and of a single chain antibody (scFv) targeted to folate-binding protein (FBP) into the N-terminus of MoMLV Env results in the reduction of the infectivity and the kinetics of entry of the MoMLV vectors. The use of soluble receptor-binding domain (sRBD), bafilomycin A1 (BafA1) and methyl-beta-cyclodextrin (MbetaC) increase the infectivity of the MoMLV vectors targeted to FBP (MoMLV-FBP) suggesting that the scFv targeted to FBP increases the threshold for fusion and might re-route entry of the targeted MoMLV-FBP vector towards an endocytic, non-productive pathway.

Education for Perspective Transformation. Women's Re-entry Programs in Community Colleges.

ERIC Educational Resources Information Center

Mezirow, Jack; Marsick, Victoria

The national field study of women's re-entry programs in community colleges reported here was done to identify factors that impeded or facilitated the progress of these programs. After an introduction, the content is presented in three sections. The first section deals with perspective transformation, the adult development process occurring in the…

Jett, Tanner and Garneau during re-entry preparations for STS-97

NASA Image and Video Library

2000-12-11

STS097-310-032 (11 December 2000) --- Astronauts (left to right) Marc Garneau, Joseph R. Tanner, both mission specialists, and Brent W. Jett, mission commander, are photographed on the flight deck of the Space Shuttle Endeavour as they prepare for re-entry. Garneau represents the Canadian Space Agency (CSA).

15 CFR 742.6 - Regional stability.

Code of Federal Regulations, 2013 CFR

2013-01-01

... under an ECCN 0Y521 entry must be re-classified under another ECCN within one calendar year from the date they are listed in Supplement No. 5 to part 774 of the EAR. If such re-classification does not occur within that period, classification under an ECCN 0Y521 entry expires, and such items are...

Advanced Concept

NASA Image and Video Library

2008-02-15

THIS IS A MODEL TEST OF THE 1ST STAGE RE-ENTRY. HEAT TESTING OF A 3% MODEL TO SUPPORT THE ARES/CLV FIRST STAGE RE-ENTRY. THIS OCCURRED AT ARNOLD AIR FORCE BASE, TENNESSEE IN SUPPORT OF THE CONSTELLATION/ARES PROJECT. THIS IMAGE IS EXTRACTED FROM A HIGH DEFINITION VIDEO FILE AND IS THE HIGHEST RESOLUTION AVAILABLE.

The IXV experience, from the mission conception to the flight results

NASA Astrophysics Data System (ADS)

Tumino, G.; Mancuso, S.; Gallego, J.-M.; Dussy, S.; Preaud, J.-P.; Di Vita, G.; Brunner, P.

2016-07-01

The atmospheric re-entry domain is a cornerstone of a wide range of space applications, ranging from reusable launcher stages developments, robotic planetary exploration, human space flight, to innovative applications such as reusable research platforms for in orbit validation of multiple space applications technologies. The Intermediate experimental Vehicle (IXV) is an advanced demonstrator which has performed in-flight experimentation of atmospheric re-entry enabling systems and technologies aspects, with significant advancements on Europe's previous flight experiences, consolidating Europe's autonomous position in the strategic field of atmospheric re-entry. The IXV mission objectives were the design, development, manufacturing, assembling and on-ground to in-flight verification of an autonomous European lifting and aerodynamically controlled reentry system, integrating critical re-entry technologies at system level. Among such critical technologies of interest, special attention was paid to aerodynamic and aerothermodynamics experimentation, including advanced instrumentation for aerothermodynamics phenomena investigations, thermal protections and hot-structures, guidance, navigation and flight control through combined jets and aerodynamic surfaces (i.e. flaps), in particular focusing on the technologies integration at system level for flight, successfully performed on February 11th, 2015.

The characteristics of registered nurses whose licenses expire: why they leave nursing and implications for retention and re-entry.

PubMed

Skillman, Susan M; Palazzo, Lorella; Hart, L Gary; Keepnews, David

2010-01-01

Little is known about RNs who drop their licenses and their potential re-entry into the nursing workforce. The results of this study provide insight into reasons nurses leave their careers and the barriers to re-entry, all important indicators of the current professional climate for nursing. While representing only one state, these findings suggest that RNs who allow their licenses to expire do so because they have reached retirement age or, among those who do not cite age as a factor, because many are unable or unwilling to work in the field. Inactive nurses who might otherwise appear to be likely candidates for re-entry into the profession may not be easily encouraged to practice nursing again without significant changes in their personal circumstances or the health care work environment. Effective ways to address current and pending RN workforce shortages include expanding RN education capacity to produce more RNs who can contribute to the workforce across the coming decades, and promote work environments in which RNs want to, and are able to, practice across a long nursing career.

Some characteristics of the glutathione cycle revealed by ionising and non-ionising electromagnetic radiation.

PubMed

Holt, J A

1995-10-01

The cyclic reaction of GSH-->GSSG-->GSH (designated R(exp) or R(e)) obeys the three specific features of life by producing energy in exponential quantities relative to time, is in effect irreversible and is inherited from generation to generation. In multicellular life, this reaction produces the energy for mitosis and is kept in controlled inactivity until needed to maintain perfection of form and function by energising mitosis. The immediate control of Re appears to be feedback process-dependent on the concentration of GSSG. Ultra high-frequency electromagnetic radiation of 434 MHz (UHF) will change Re from inactive to active and, in so doing, it causes resonance and/or fluorescence of the glutathione cycle which changes its radiosensitivity. Re is the primary direct target of ionising radiation and produces the energy for mitosis. Clinical observations suggest that, in the normal cell, Re is inactive and is not killed by 3 x 2700 rads or 6 x 1650 rads yet, when active, its sensitivity value (DO) is approximately 160 rads. Using the standard radiobiological equation of response to ionising radiation, it can be deduced that radiosensitive cancers have two or three Re units active per cell and radioresistance increases in proportion to the number of potentially active Re units per cell. Re appears to be the main cause of cancers' increased conductivity of electricity compared with normal tissue. In cancer therapy, UHF is the best radiosensitiser ever discovered (up to two or more decades). Re is also intelligent compared with non-exponential reactions but cannot be the basis of intellectual brain functions which must be based on non-electrical chemical processes.

Origin of bistability underlying mammalian cell cycle entry

PubMed Central

Yao, Guang; Tan, Cheemeng; West, Mike; Nevins, Joseph R; You, Lingchong

2011-01-01

Precise control of cell proliferation is fundamental to tissue homeostasis and differentiation. Mammalian cells commit to proliferation at the restriction point (R-point). It has long been recognized that the R-point is tightly regulated by the Rb–E2F signaling pathway. Our recent work has further demonstrated that this regulation is mediated by a bistable switch mechanism. Nevertheless, the essential regulatory features in the Rb–E2F pathway that create this switching property have not been defined. Here we analyzed a library of gene circuits comprising all possible link combinations in a simplified Rb–E2F network. We identified a minimal circuit that is able to generate robust, resettable bistability. This minimal circuit contains a feed-forward loop coupled with a mutual-inhibition feedback loop, which forms an AND-gate control of the E2F activation. Underscoring its importance, experimental disruption of this circuit abolishes maintenance of the activated E2F state, supporting its importance for the bistability of the Rb–E2F system. Our findings suggested basic design principles for the robust control of the bistable cell cycle entry at the R-point. PMID:21525871

Numerical and experimental study of the thermal degradation process during the atmospheric re-entry of a TiAl6V4 tank

NASA Astrophysics Data System (ADS)

Prévereaud, Y.; Vérant, J.-L.; Balat-Pichelin, M.; Moschetta, J.-M.

2016-05-01

To answer the question of space debris survivability during atmospheric entry ONERA uses its software named MUSIC/FAST. So, the first part of this paper is dedicated to the presentation of the ONERA tool and its validation by comparison with flight data and CFD computations. However, the influence of oxidation on the thermal degradation process and material properties in atmospheric entry conditions is still unknown. A second step is then devoted to the presentation of an experimental campaign investigating TA6V oxidation in atmospheric entry conditions, as the most of the debris found on ground are made of this material. Experiments have been realized using the MESOX facility implemented at the 6 kW solar furnace in PROMES-CNRS laboratory. Finally, an application of MUSIC/FAST is proposed on the atmospheric re-entry of a generic TA6V tank. Aiming at degradation assessment, a sensitive study to initial conditions is conducted. To complete computational analysis regarding degradation process by melting, a numerical analysis of the influence of oxidation on the thermal wall degradation during the tank atmospheric re-entry is presented as well.

pRb phosphorylation regulates the proliferation of supporting cells in gentamicin-damaged neonatal avian utricle.

PubMed

Wu, Jingfang; Sun, Shan; Li, Wenyan; Chen, Yan; Li, Huawei

2014-10-01

The ability of nonmammalian vertebrates to regenerate hair cells (HCs) after damage-induced HC loss has stimulated and inspired research in the field of HC regeneration. The protein pRb encoded by retinoblastoma gene Rb1 forces sensory progenitor cells to exit cell cycle and maintain differentiated HCs and supporting cells (SCs) in a quiescent state. pRb function is regulated by phosphorylation through the MEK/ERK or the pRb/Raf-1 signaling pathway. In our previous study, we have shown that pRb phosphorylation is crucial for progenitor cell proliferation and survival during the early embryonic stage of avian otocyst sensory epithelium development. However, in damaged avian utricle, the role of pRb in regulating the cell cycling of SCs or HCs regeneration still remains unclear. To further elucidate the function of pRb phosphorylation on SCs re-entering the cell cycle triggered by gentamycin-induced HCs damage, we isolated neonatal chicken utricles and treated them with the MEK inhibitor U0126 or the pRb/Raf-1 inhibitor RRD-251, respectively in vitro. We found that after gentamycin-induced HCs damage, pRb phosphorylation is important for the quiescent SCs re-entering the cell cycle in the neonatal chicken utricle. In addition, the proliferation of SCs decreased in a dose-dependent manner in response to both U0126 and RRD-251, which indicates that both the MEK/ERK and the pRb/Raf-1 signaling pathway play important roles in pRb phosphorylation in damaged neonatal chicken utricle. Together, these findings on the function of pRb in damaged neonatal chicken utricle improve our understanding of the regulation of the cell cycle of SCs after HCs loss and may shed light on the mammalian HC regeneration from SCs in damaged organs.

A Microbial Avenue to Cell Cycle Control in the Plant Superkingdom[C][W][OPEN

PubMed Central

Tulin, Frej; Cross, Frederick R.

2014-01-01

Research in yeast and animals has resulted in a well-supported consensus model for eukaryotic cell cycle control. The fit of this model to early diverging eukaryotes, such as the plant kingdom, remains unclear. Using the green alga Chlamydomonas reinhardtii, we developed an efficient pipeline, incorporating robotics, semiautomated image analysis, and deep sequencing, to molecularly identify >50 genes, mostly conserved in higher plants, specifically required for cell division but not cell growth. Mutated genes include the cyclin-dependent kinases CDKA (resembling yeast and animal Cdk1) and the plant-specific CDKB. The Chlamydomonas cell cycle consists of a long G1 during which cells can grow >10-fold, followed by multiple rapid cycles of DNA replication and segregation. CDKA and CDKB execute nonoverlapping functions: CDKA promotes transition between G1 and entry into the division cycle, while CDKB is essential specifically for spindle formation and nuclear division, but not for DNA replication, once CDKA-dependent initiation has occurred. The anaphase-promoting complex is required for similar steps in the Chlamydomonas cell cycle as in Opisthokonts; however, the spindle assembly checkpoint, which targets the APC in Opisthokonts, appears severely attenuated in Chlamydomonas, based on analysis of mutants affecting microtubule function. This approach allows unbiased integration of the consensus cell cycle control model with innovations specific to the plant lineage. PMID:25336509

Cardiovascular effects of anti-G suit and cooling garment during space shuttle re-entry and landing.

PubMed

Perez, Sondra A; Charles, John B; Fortner, G William; Hurst, Victor; Meck, Janice V

2003-07-01

Many cardiovascular changes associated with spaceflight reduce the ability of the cardiovascular system to oppose gravity on return to Earth, leaving astronauts susceptible to orthostatic hypotension during re-entry and landing. Consequently, an anti-G suit was developed to protect arterial pressure during re-entry. A liquid cooling garment (LCG) was then needed to alleviate the thermal stress resulting from use of the launch and entry suit. We studied 34 astronauts on 22 flights (4-16 d). Subjects were studied 10 d before launch and on landing day. Preflight, crewmembers were suited with their anti-G suits set to the intended inflation for re-entry. Three consecutive measurements of heart rate and arterial pressure were obtained while seated and then again while standing. Three subjects who inflated the anti-G suits also donned the LCG for landing. Arterial pressure and heart rate were measured every 5 min during the de-orbit maneuver, through maximum G-loading (max-G) and touch down (TD). After TD, crew-members again initiated three seated measurements followed by three standing measurements. Astronauts with inflated anti-G suits had higher arterial pressure than those who did not have inflated anti-G suits during re-entry and landing (133.1 +/- 2.5/76.1 +/- 2.1 vs. 128.3 +/- 4.2/79.3 +/- 2.9, de-orbit; 157.3 +/- 4.5/102.1 +/- 3.6 vs. 145.2 +/- 10.5/95.7 + 5.5, max-G; 159.6 +/- 3.9/103.7 +/- 3.3 vs. 134.1 +/- 5.1/85.7 +/- 3.1, TD). In the group with inflated anti-G suits, those who also wore the LCG exhibited significantly lower heart rates than those who did not (75.7 +/- 11.5 vs. 86.5 +/- 6.2, de-orbit; 79.5 +/- 24.8 vs. 112.1 +/- 8.7, max-G; 84.7 +/- 8.0 vs. 110.5 +/- 7.9, TD). The anti-G suit is effective in supporting arterial pressure. The addition of the LCG lowers heart rate during re-entry.

Cardiovascular effects of anti-G suit and cooling garment during space shuttle re-entry and landing

NASA Technical Reports Server (NTRS)

Perez, Sondra A.; Charles, John B.; Fortner, G. William; Hurst, Victor 4th; Meck, Janice V.

2003-01-01

BACKGROUND: Many cardiovascular changes associated with spaceflight reduce the ability of the cardiovascular system to oppose gravity on return to Earth, leaving astronauts susceptible to orthostatic hypotension during re-entry and landing. Consequently, an anti-G suit was developed to protect arterial pressure during re-entry. A liquid cooling garment (LCG) was then needed to alleviate the thermal stress resulting from use of the launch and entry suit. METHODS: We studied 34 astronauts on 22 flights (4-16 d). Subjects were studied 10 d before launch and on landing day. Preflight, crewmembers were suited with their anti-G suits set to the intended inflation for re-entry. Three consecutive measurements of heart rate and arterial pressure were obtained while seated and then again while standing. Three subjects who inflated the anti-G suits also donned the LCG for landing. Arterial pressure and heart rate were measured every 5 min during the de-orbit maneuver, through maximum G-loading (max-G) and touch down (TD). After TD, crew-members again initiated three seated measurements followed by three standing measurements. RESULTS: Astronauts with inflated anti-G suits had higher arterial pressure than those who did not have inflated anti-G suits during re-entry and landing (133.1 +/- 2.5/76.1 +/- 2.1 vs. 128.3 +/- 4.2/79.3 +/- 2.9, de-orbit; 157.3 +/- 4.5/102.1 +/- 3.6 vs. 145.2 +/- 10.5/95.7 + 5.5, max-G; 159.6 +/- 3.9/103.7 +/- 3.3 vs. 134.1 +/- 5.1/85.7 +/- 3.1, TD). In the group with inflated anti-G suits, those who also wore the LCG exhibited significantly lower heart rates than those who did not (75.7 +/- 11.5 vs. 86.5 +/- 6.2, de-orbit; 79.5 +/- 24.8 vs. 112.1 +/- 8.7, max-G; 84.7 +/- 8.0 vs. 110.5 +/- 7.9, TD). CONCLUSIONS: The anti-G suit is effective in supporting arterial pressure. The addition of the LCG lowers heart rate during re-entry.

Re-entry and related predictors among HIV-infected clients receiving methadone maintenance treatment in Guangdong province, China.

PubMed

Luo, Xiaofeng; Gong, Xiao; Zhao, Peizhen; Zou, Xia; Chen, Wen; Ling, Li

2017-07-24

This study examined the re-entry characteristics and related predictors among HIV-infected methadone maintenance treatment (MMT) clients in Guangdong, China. Data on HIV-infected MMT clients was obtained from the clinic MMT registration system in Guangdong. Of the 653 participants, only 9.0% remained in the MMT program until the end of the study. For the drop-outs, 70.0% returned to MMT at least once by the end of the study. Re-entry was independently associated with marital status (OR never married = 2.24, 95% CI: 1.02-4.93; OR married currently = 2.34, 95% CI: 1.05-5.22), being unemployed (OR = 1.92, 95% CI: 1.12-3.27), lower positive percentages of urine tests (OR <40% = 4.08, 95% CI: 2.21-7.54; OR 40%-80% = 2.52, 95% CI: 1.39-4.56), higher maintenance doses (OR = 3.78, 95% CI: 2.21-7.54)and poorer MMT attendance percentages (OR <20% = 282.02, 95% CI: 62.75-1268.11; OR20-49% = 20.75, 95% CI: 10.52-40.93; OR50-79% = 6.07, 95% CI: 3.44-10.73). A higher re-entry frequency was independently associated with lower education level (ORjunior high school = 0.49, 95% CI: 0.26-0.93), average drug use times less than twice (OR = 0.64, 95% CI: 0.41-1.00), lower positive percentages of urine tests (OR = 0.39, 95% CI: 0.22-0.70) and poorer percentages of MMT attendance (OR<20% = 7.24, 95% CI: 2.99-17.55; OR20-49% = 14.30, 95% CI: 5.94-34.42; OR50-79% = 6.15, 95% CI: 2.55-14.85). Re-entry and repeated re-entry were prevalent among HIV-infected MMT clients in Guangdong, underscoring the urgent needs of tailored interventions and health education programs for this population.

One-year outcomes after successful chronic total occlusion percutaneous coronary intervention: The impact of dissection re-entry techniques.

PubMed

Wilson, W M; Walsh, S J; Bagnall, A; Yan, A T; Hanratty, C G; Egred, M; Smith, E; Oldroyd, K G; McEntegart, M; Irving, J; Douglas, H; Strange, J; Spratt, J C

2017-11-01

We aimed to determine clinical outcomes 1 year after successful chronic total occlusion (CTO) PCI and, in particular, whether use of dissection and re-entry strategies affects clinical outcomes. Hybrid approaches have increased the procedural success of CTO percutaneous coronary intervention (PCI) but longer-term outcomes are unknown, particularly in relation to dissection and re-entry techniques. Data were collected for consecutive CTO PCIs performed by hybrid-trained operators from 7 United Kingdom (UK) centres between 2012 and 2014. The primary endpoint (death, myocardial infarction, unplanned target vessel revascularization) was measured at 12 months along with angina status. One-year follow up data were available for 96% of successful cases (n = 805). In total, 85% of patients had a CCS angina class of 2-4 prior to CTO PCI. Final successful procedural strategy was antegrade wire escalation 48%; antegrade dissection and re-entry (ADR) 21%; retrograde wire escalation 5%; retrograde dissection and re-entry (RDR) 26%. Overall, 47% of CTOs were recanalized using dissection and re-entry strategies. During a mean follow up of 11.5 ± 3.8 months, the primary endpoint occurred in 8.6% (n = 69) of patients (10.3% (n = 39/375) in DART group and 7.0% (n = 30/430) in wire-based cases). The majority of patients (88%) had no or minimal angina (CCS class 0 or 1). ADR and RDR were used more frequently in more complex cases with greater disease burden, however, the only independent predictor of the primary endpoint was lesion length. CTO PCI in complex lesions using the hybrid approach is safe, effective and has a low one-year adverse event rate. The method used to recanalize arteries was not associated with adverse outcomes. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

Multicentre experience with the BridgePoint devices to facilitate recanalisation of chronic total coronary occlusions through controlled subintimal re-entry.

PubMed

Werner, Gerald S; Schofer, Joachim; Sievert, Horst; Kugler, Chad; Reifart, Nicolaus J

2011-06-01

The major challenge for the interventional treatment of chronic total coronary occlusion (CTO) is a low primary success rate. A common problem is the passage of the recanalisation wire into a subintimal position. New devices, which were evaluated in the first multicentre study in CTOs resistant to a conventional wire approach, may help to facilitate a controlled re-entry into the true lumen. The aim of this study was to assess the safety and efficacy of this approach, with successful true lumen distal wire passage as the primary endpoint. Forty-two patients were enrolled in four centres with high expertise in PCI for CTOs. All CTOs were of at least three months duration, and were initially attempted with dedicated recanalisation wires. After failure to pass or creation of a subintimal dissection, the BridgePoint devices were applied, consisting of a ball-tipped catheter (CrossBoss) to pass the proximal occlusion cap, and a flat-shaped balloon catheter (Stingray catheter) to be inflated within the subintimal space to guide the re-entry into the true vessel lumen with a special wire (Stingray guidewire). The primary endpoint was met in 67% of all patients. A higher success rate seemed to be possible when all devices were used in sequenced beginning with the CrossBoss, and in the case of a subintimal passage, followed by the Stingray. True lumen re-entry failed because of the loss of distally contrast filling and thus loss of a target for re-entry, and by a failure to advance the Stingray balloon far enough distal and parallel to the distal lumen. There were no severe device related complications. In patients with complex CTOs referred to dedicated centres with high experience in CTOs, these results demonstrate the potential of a guided re-entry from a subintimal wire position by use of the BridgePoint devices.

IXV re-entry demonstrator: Mission overview, system challenges and flight reward

NASA Astrophysics Data System (ADS)

Angelini, Roberto; Denaro, Angelo

2016-07-01

The Intermediate eXperimental Vehicle (IXV) is an advanced re-entry demonstrator vehicle aimed to perform in-flight experimentation of atmospheric re-entry enabling systems and technologies. The IXV integrates key technologies at the system level, with significant advancements on Europe's previous flying test-beds. The project builds on previous achievements at system and technology levels, and provides a unique and concrete way of establishing and consolidating Europe's autonomous position in the strategic field of atmospheric re-entry. The IXV mission and system objectives are the design, development, manufacturing, assembling and on-ground to in-flight verification of an autonomous European lifting and aerodynamically controlled reentry system, integrating critical re-entry technologies at system level. Among such critical technologies of interest, special attention is paid to aerodynamic and aerothermodynamics experimentation, including advanced instrumentation for aerothermodynamics phenomena investigations, thermal protections and hot-structures, guidance, navigation and flight control through combined jets and aerodynamic surfaces (i.e. flaps), in particular focusing on the technologies integration at system level for flight. Following the extensive detailed design, manufacturing, qualification, integration and testing of the flight segment and ground segment elements, IXV has performed a full successful flight on February 11th 2015. After the launch with the VEGA launcher form the CSG spaceport in French Guyana, IXV has performed a full nominal mission ending with a successful splashdown in the Pacific Ocean. During Flight Phase, the IXV space and ground segments worked perfectly, implementing the whole flight program in line with the commanded maneuvers and trajectory prediction, performing an overall flight of 34.400 km including 7.600 km with hot atmospheric re-entry in automatic guidance, concluding with successful precision landing at a distance of ~1 km from the target, including the wind drift acting on the parachute from an altitude of 4.5 km.

BeatBox-HPC simulation environment for biophysically and anatomically realistic cardiac electrophysiology.

PubMed

Antonioletti, Mario; Biktashev, Vadim N; Jackson, Adrian; Kharche, Sanjay R; Stary, Tomas; Biktasheva, Irina V

2017-01-01

The BeatBox simulation environment combines flexible script language user interface with the robust computational tools, in order to setup cardiac electrophysiology in-silico experiments without re-coding at low-level, so that cell excitation, tissue/anatomy models, stimulation protocols may be included into a BeatBox script, and simulation run either sequentially or in parallel (MPI) without re-compilation. BeatBox is a free software written in C language to be run on a Unix-based platform. It provides the whole spectrum of multi scale tissue modelling from 0-dimensional individual cell simulation, 1-dimensional fibre, 2-dimensional sheet and 3-dimensional slab of tissue, up to anatomically realistic whole heart simulations, with run time measurements including cardiac re-entry tip/filament tracing, ECG, local/global samples of any variables, etc. BeatBox solvers, cell, and tissue/anatomy models repositories are extended via robust and flexible interfaces, thus providing an open framework for new developments in the field. In this paper we give an overview of the BeatBox current state, together with a description of the main computational methods and MPI parallelisation approaches.

Dynamics of re-constitution of the human nuclear proteome after cell division is regulated by NLS-adjacent phosphorylation

PubMed Central

Róna, Gergely; Borsos, Máté; Ellis, Jonathan J; Mehdi, Ahmed M; Christie, Mary; Környei, Zsuzsanna; Neubrandt, Máté; Tóth, Judit; Bozóky, Zoltán; Buday, László; Madarász, Emília; Bodén, Mikael; Kobe, Bostjan; Vértessy, Beáta G

2014-01-01

Phosphorylation by the cyclin-dependent kinase 1 (Cdk1) adjacent to nuclear localization signals (NLSs) is an important mechanism of regulation of nucleocytoplasmic transport. However, no systematic survey has yet been performed in human cells to analyze this regulatory process, and the corresponding cell-cycle dynamics have not yet been investigated. Here, we focused on the human proteome and found that numerous proteins, previously not identified in this context, are associated with Cdk1-dependent phosphorylation sites adjacent to their NLSs. Interestingly, these proteins are involved in key regulatory events of DNA repair, epigenetics, or RNA editing and splicing. This finding indicates that cell-cycle dependent events of genome editing and gene expression profiling may be controlled by nucleocytoplasmic trafficking. For in-depth investigations, we selected a number of these proteins and analyzed how point mutations, expected to modify the phosphorylation ability of the NLS segments, perturb nucleocytoplasmic localization. In each case, we found that mutations mimicking hyper-phosphorylation abolish nuclear import processes. To understand the mechanism underlying these phenomena, we performed a video microscopy-based kinetic analysis to obtain information on cell-cycle dynamics on a model protein, dUTPase. We show that the NLS-adjacent phosphorylation by Cdk1 of human dUTPase, an enzyme essential for genomic integrity, results in dynamic cell cycle-dependent distribution of the protein. Non-phosphorylatable mutants have drastically altered protein re-import characteristics into the nucleus during the G1 phase. Our results suggest a dynamic Cdk1-driven mechanism of regulation of the nuclear proteome composition during the cell cycle. PMID:25483092

On-Ground Casualty Risk Reduction by Structural Design for Demise

NASA Astrophysics Data System (ADS)

Lemmens, Stijn; Krag, Holger; Funke, Quirin

In recent years, awareness concerning the risk posed by un-controlled re-entering spacecraft on ground has increased. Some re-entry events such as ESA's GOCE in 2013 and NASA's UARS appeared prominent in international media. Space agencies and nations, in cooperation within the Inter-Agency Space Debris Coordination Committee (IADC), have established a requirements to limited the on-ground risk for future missions. To meet the requirements, the amount of debris falling back on Earth has to be limited in number, mass and size. Design for demise (D4D) refers to all measures taken in the design of a space object to increase the potential for demise of the object and its components during re-entry. SCARAB (Spacecraft Atmospheric Re-entry and Break-Up) is ESA's high-fidelity tool which analyses the thermal and structural effects of atmospheric re-entry on spacecraft in a finite-element approach. For this study, a model of a representative satellite is developed in Scarab to serve as test-bed for D4D analysis on a structural level. The model is used as starting point for different D4D approaches based on increasing the exposure of the satellite components to the aero-thermal environment, as a way to speed up the demise. Statistical bootstrapping is applied to the resulting on-ground fragment lists in order to compare the different re-entry scenarios and to determine the uncertainties of the results. Moreover, the bootstrap results can be used to analyse the casualty risk estimator from a theoretical point of view. The risk reductions for the analysed D4D techniques are presented w.r.t. the reference scenario for the modelled representative satellite.

Factors associated with dropout among patients in opioid maintenance treatment (OMT) and predictors of re-entry. A national registry-based study.

PubMed

Bukten, Anne; Skurtveit, Svetlana; Waal, Helge; Clausen, Thomas

2014-10-01

Retention in treatment is often highlighted as one of the key indicators of success in opioid maintenance treatment (OMT). To identify factors associated with long-term retention in opioid maintenance treatment and to analyse predictors of subsequent treatment episodes. Treatment retention and re-entry were examined for a national cohort of patients admitted to OMT in Norway in the period 1997-2003. Multivariate Cox regression models were used to investigate factors associated with treatment dropout 18months after treatment entry. The 18month retention rate among patients admitted to OMT in Norway (n=2431) was 65.8% (n=1599). Dropout from OMT within 18months was associated with younger age (HR 0.97 [0.96-0.98]), high levels of general pre-treatment criminal offences (HR 1.66 [1.32-2.09]) and having drug-related offences during the 30days prior to dropout (HR 1.80 [1.36-2.38]). Of the patients who dropped out (n=832), 42.7% (n=355) were re-engaged in subsequent treatment episodes. Pre-treatment criminal offences were associated with increased odds for treatment re-entry, whereas being younger and having drug-related offences during the first OMT episode were associated with lower odds for re-engagement in OMT. Gender was not associated with treatment dropout and re-entry. High levels of pre-treatment criminal offences and drug offences during the 30days prior to dropout were associated with treatment dropout. Efforts to increase support services to these patients may contribute to higher rates of retention in OMT. Copyright © 2014 Elsevier Ltd. All rights reserved.

Purification and stability characterization of a cell regulatory sialoglycopeptide inhibitor

NASA Technical Reports Server (NTRS)

Moos, P. J.; Fattaey, H. K.; Johnson, T. C.; Spooner, B. S. (Principal Investigator)

1995-01-01

Previous attempts to physically separate the cell cycle inhibitory and protease activities in preparations of a purified cell regulatory sialoglycopeptide (CeReS) inhibitor were largely unsuccessful. Gradient elution of the inhibitor preparation from a DEAE HPLC column separated the cell growth inhibitor from the protease, and the two activities have been shown to be distinct and non-overlapping. The additional purification increased the specific biological activity of the CeReS preparation by approximately two-fold. The major inhibitory fraction that eluted from the DEAE column was further analyzed by tricine-SDS-PAGE and microbore reverse phase HPLC and shown to be homogeneous in nature. Two other fractions separated by DEAE HPLC, also devoid of protease activity, were shown to be inhibitory to cell proliferation and most likely represented modified relatives of the CeReS inhibitor. The highly purified CeReS was chemically characterized for amino acid and carbohydrate composition and the role of the carbohydrate in cell proliferation inhibition, stability, and protease resistance was assessed.

Regulation of DNA synthesis and the cell cycle in human prostate cancer cells and lymphocytes by ovine uterine serpin

PubMed Central

Padua, Maria B; Hansen, Peter J

2008-01-01

Background Uterine serpins are members of the serine proteinase inhibitor superfamily. Like some other serpins, these proteins do not appear to be functional proteinase inhibitors. The most studied member of the group, ovine uterine serpin (OvUS), inhibits proliferation of several cell types including activated lymphocytes, bovine preimplantation embryos, and cell lines for lymphoma, canine primary osteosarcoma and human prostate cancer (PC-3) cells. The goal for the present study was to evaluate the mechanism by which OvUS inhibits cell proliferation. In particular, it was tested whether inhibition of DNA synthesis in PC-3 cells involves cytotoxic actions of OvUS or the induction of apoptosis. The effect of OvUS in the production of the autocrine and angiogenic cytokine interleukin (IL)-8 by PC-3 cells was also determined. Finally, it was tested whether OvUS blocks specific steps in the cell cycle using both PC-3 cells and lymphocytes. Results Recombinant OvUS blocked proliferation of PC-3 cells at concentrations as low as 8 μg/ml as determined by measurements of [3H]thymidine incorporation or ATP content per well. Treatment of PC-3 cells with OvUS did not cause cytotoxicity or apoptosis or alter interleukin-8 secretion into medium. Results from flow cytometry experiments showed that OvUS blocked the entry of PC-3 cells into S phase and the exit from G2/M phase. In addition, OvUS blocked entry of lymphocytes into S phase following activation of proliferation with phytohemagglutinin. Conclusion Results indicate that OvUS acts to block cell proliferation through disruption of the cell cycle dynamics rather than induction of cytotoxicity or apoptosis. The finding that OvUS can regulate cell proliferation makes this one of only a few serpins that function to inhibit cell growth. PMID:18218135

DREAMs make plant cells to cycle or to become quiescent.

PubMed

Magyar, Zoltán; Bögre, László; Ito, Masaki

2016-12-01

Cell cycle phase specific oscillation of gene transcription has long been recognized as an underlying principle for ordered processes during cell proliferation. The G1/S-specific and G2/M-specific cohorts of genes in plants are regulated by the E2F and the MYB3R transcription factors. Mutant analysis suggests that activator E2F functions might not be fully required for cell cycle entry. In contrast, the two activator-type MYB3Rs are part of positive feedback loops to drive the burst of mitotic gene expression, which is necessary at least to accomplish cytokinesis. Repressor MYB3Rs act outside the mitotic time window during cell cycle progression, and are important for the shutdown of mitotic genes to impose quiescence in mature organs. The two distinct classes of E2Fs and MYB3Rs together with the RETINOBLATOMA RELATED are part of multiprotein complexes that may be evolutionary related to what is known as DREAM complex in animals. In plants, there are multiple such complexes with distinct compositions and functions that may be involved in the coordinated cell cycle and developmental regulation of E2F targets and mitotic genes. Copyright © 2016 Elsevier Ltd. All rights reserved.

Ubiquitin in Influenza Virus Entry and Innate Immunity.

PubMed

Rudnicka, Alina; Yamauchi, Yohei

2016-10-24

Viruses are obligatory cellular parasites. Their mission is to enter a host cell, to transfer the viral genome, and to replicate progeny whilst diverting cellular immunity. The role of ubiquitin is to regulate fundamental cellular processes such as endocytosis, protein degradation, and immune signaling. Many viruses including influenza A virus (IAV) usurp ubiquitination and ubiquitin-like modifications to establish infection. In this focused review, we discuss how ubiquitin and unanchored ubiquitin regulate IAV host cell entry, and how histone deacetylase 6 (HDAC6), a cytoplasmic deacetylase with ubiquitin-binding activity, mediates IAV capsid uncoating. We also discuss the roles of ubiquitin in innate immunity and its implications in the IAV life cycle.

Ubiquitin in Influenza Virus Entry and Innate Immunity

PubMed Central

Rudnicka, Alina; Yamauchi, Yohei

2016-01-01

Viruses are obligatory cellular parasites. Their mission is to enter a host cell, to transfer the viral genome, and to replicate progeny whilst diverting cellular immunity. The role of ubiquitin is to regulate fundamental cellular processes such as endocytosis, protein degradation, and immune signaling. Many viruses including influenza A virus (IAV) usurp ubiquitination and ubiquitin-like modifications to establish infection. In this focused review, we discuss how ubiquitin and unanchored ubiquitin regulate IAV host cell entry, and how histone deacetylase 6 (HDAC6), a cytoplasmic deacetylase with ubiquitin-binding activity, mediates IAV capsid uncoating. We also discuss the roles of ubiquitin in innate immunity and its implications in the IAV life cycle. PMID:27783058

Establishment of a Re-Entry Model to Reduce Recidivism Among Court Ward Students.

ERIC Educational Resources Information Center

Kammuller, Kenneth C.

The development and implementation of a Re-Entry Model designed to facilitate and monitor the adjustment of high school students returned to public school from a county commitment facility is described. Transition and follow-up procedures implemented through the model with a liaison teacher at the commitment facility school designated as the…

Career Assessment, Remediation, Education, Employment, and Re-entry Program (CAREER). El Paso Community College Career Grant. Final Report.

ERIC Educational Resources Information Center

LaFleur, Carol A.

Objectives of the Career Assessment, Remediation, Education, Employment, and Re-entry (CAREER) project were to establish a series of intensive, short-term job training programs using competency-based instruction to serve Hispanic persons who were economically disadvantaged, displaced, unemployed, or underemployed, as well as Hispanic females who…

50 CFR 300.114 - Dealer permits and preapproval.

Code of Federal Regulations, 2013 CFR

2013-10-01

... receipt, importation, or re-export. Dealers must supply the U.S. Customs 7501 entry number at least three...) General. (1) A dealer intending to import or re-export AMLR must obtain an AMLR dealer permit valid for... preapproval issued under § 300.114(a)(1). AMLRs may not be released for entry into the United States unless...

50 CFR 300.114 - Dealer permits and preapproval.

Code of Federal Regulations, 2010 CFR

2010-10-01

... receipt, importation, or re-export. Dealers must supply the U.S. Customs 7501 entry number at least three...) General. (1) A dealer intending to import or re-export AMLR must obtain an AMLR dealer permit valid for... preapproval issued under § 300.114(a)(1). AMLRs may not be released for entry into the United States unless...

7 CFR 1530.107 - Bond or letter of credit requirements.

Code of Federal Regulations, 2013 CFR

2013-01-01

... SERVICE, DEPARTMENT OF AGRICULTURE THE REFINED SUGAR RE-EXPORT PROGRAM, THE SUGAR CONTAINING PRODUCTS RE..., which meets the criteria set forth in this section. (b) The bond or letter of credit may cover entries made either during the period of time specified in the bond (a term bond) or for a specified entry (a...

Exploring Efficacy in Negotiating Support: Women Re-Entry Students in Higher Education

ERIC Educational Resources Information Center

Filipponi-Berardinelli, Josephine Oriana

2013-01-01

The existing literature on women re-entry students reveals that women students concurrently struggle with family, work, and sometimes health issues. Women students often do not receive adequate support from their partners or from other sources in helping manage the multiple roles that compete for their time, and often face constraints that affect…

7 CFR 1530.107 - Bond or letter of credit requirements.

Code of Federal Regulations, 2012 CFR

2012-01-01

... SERVICE, DEPARTMENT OF AGRICULTURE THE REFINED SUGAR RE-EXPORT PROGRAM, THE SUGAR CONTAINING PRODUCTS RE..., which meets the criteria set forth in this section. (b) The bond or letter of credit may cover entries made either during the period of time specified in the bond (a term bond) or for a specified entry (a...

7 CFR 1530.107 - Bond or letter of credit requirements.

Code of Federal Regulations, 2014 CFR

2014-01-01

... SERVICE, DEPARTMENT OF AGRICULTURE THE REFINED SUGAR RE-EXPORT PROGRAM, THE SUGAR CONTAINING PRODUCTS RE..., which meets the criteria set forth in this section. (b) The bond or letter of credit may cover entries made either during the period of time specified in the bond (a term bond) or for a specified entry (a...

50 CFR 300.114 - Dealer permits and preapproval.

Code of Federal Regulations, 2011 CFR

2011-10-01

... receipt, importation, or re-export. Dealers must supply the U.S. Customs 7501 entry number at least three...) General. (1) A dealer intending to import or re-export AMLR must obtain an AMLR dealer permit valid for... preapproval issued under § 300.114(a)(1). AMLRs may not be released for entry into the United States unless...

50 CFR 300.114 - Dealer permits and preapproval.

Code of Federal Regulations, 2014 CFR

2014-10-01

... receipt, importation, or re-export. Dealers must supply the U.S. Customs 7501 entry number at least three...) General. (1) A dealer intending to import or re-export AMLR must obtain an AMLR dealer permit valid for... preapproval issued under § 300.114(a)(1). AMLRs may not be released for entry into the United States unless...

50 CFR 300.114 - Dealer permits and preapproval.

Code of Federal Regulations, 2012 CFR

2012-10-01

... receipt, importation, or re-export. Dealers must supply the U.S. Customs 7501 entry number at least three...) General. (1) A dealer intending to import or re-export AMLR must obtain an AMLR dealer permit valid for... preapproval issued under § 300.114(a)(1). AMLRs may not be released for entry into the United States unless...

7 CFR 1530.107 - Bond or letter of credit requirements

Code of Federal Regulations, 2011 CFR

2011-01-01

... SERVICE, DEPARTMENT OF AGRICULTURE THE REFINED SUGAR RE-EXPORT PROGRAM, THE SUGAR CONTAINING PRODUCTS RE..., which meets the criteria set forth in this section. (b) The bond or letter of credit may cover entries made either during the period of time specified in the bond (a term bond) or for a specified entry (a...

Advanced Concept

NASA Image and Video Library

2008-02-15

THIS IS A TEST OF THE 1ST STAGE RE-ENTRY VEHICLE. HEAT TESTING OF A 3% MODEL TO SUPPORT THE ARES/ CLV FIRST STAGE RE-ENTRY. THIS TEST OCCURRED AT ARNOLD AIR FORCE BASE, TENNESSEE. THIS TESTING SUPPORTS THE DEVELOPMENT OF THE CONSTELLATION/ARES PROJECT. THIS IMAGE IS EXTRACTED FROM A HIGH DEFINITION VIDEO FILE AND IS THE HIGHEST RESOLUTION AVAILABLE.

ATV-3 undock from ISS

NASA Image and Video Library

2012-09-28

ISS033-E-007940 (28 Sept. 2012) --- European Space Agency's "Edoardo Amaldi" Automated Transfer Vehicle-3 (ATV-3) begins its relative separation from the International Space Station during the Expedition 33 mission. The ATV-3 undocked from the aft port of the Zvezda Service Module at 5:44 p.m. (EDT) on Sept. 28, 2012. The ATV-3 is scheduled to deorbit on Oct. 2 for a fiery re-entry over the Pacific Ocean that will destroy the trash-filled spacecraft. Inside the ATV-3 is the Re-Entry Breakup Recorder that will record various data such as temperature, pressure and speed as the resupply craft burns up during its return to Earth. Experts will use that data to design safer and more predictable destructive re-entry techniques.

ATV-3 undock from ISS

NASA Image and Video Library

2012-09-28

ISS033-E-008016 (28 Sept. 2012) --- European Space Agency's "Edoardo Amaldi" Automated Transfer Vehicle-3 (ATV-3) begins its relative separation from the International Space Station during the Expedition 33 mission. The ATV-3 undocked from the aft port of the Zvezda Service Module at 5:44 p.m. (EDT) on Sept. 28, 2012. The ATV-3 is scheduled to deorbit on Oct. 2 for a fiery re-entry over the Pacific Ocean that will destroy the trash-filled spacecraft. Inside the ATV-3 is the Re-Entry Breakup Recorder that will record various data such as temperature, pressure and speed as the resupply craft burns up during its return to Earth. Experts will use that data to design safer and more predictable destructive re-entry techniques.

ATV-3 undock from ISS

NASA Image and Video Library

2012-09-28

ISS033-E-007980 (28 Sept. 2012) --- European Space Agency's "Edoardo Amaldi" Automated Transfer Vehicle-3 (ATV-3) begins its relative separation from the International Space Station during the Expedition 33 mission. The ATV-3 undocked from the aft port of the Zvezda Service Module at 5:44 p.m. (EDT) on Sept. 28, 2012. The ATV-3 is scheduled to deorbit on Oct. 2 for a fiery re-entry over the Pacific Ocean that will destroy the trash-filled spacecraft. Inside the ATV-3 is the Re-Entry Breakup Recorder that will record various data such as temperature, pressure and speed as the resupply craft burns up during its return to Earth. Experts will use that data to design safer and more predictable destructive re-entry techniques.

ATV-3 undock from ISS

NASA Image and Video Library

2012-09-28

ISS033-E-007915 (28 Sept. 2012) --- European Space Agency's "Edoardo Amaldi" Automated Transfer Vehicle-3 (ATV-3) begins its relative separation from the International Space Station during the Expedition 33 mission. The ATV-3 undocked from the aft port of the Zvezda Service Module at 5:44 p.m. (EDT) on Sept. 28, 2012. The ATV-3 is scheduled to deorbit on Oct. 2 for a fiery re-entry over the Pacific Ocean that will destroy the trash-filled spacecraft. Inside the ATV-3 is the Re-Entry Breakup Recorder that will record various data such as temperature, pressure and speed as the resupply craft burns up during its return to Earth. Experts will use that data to design safer and more predictable destructive re-entry techniques.

ATV-3 undock from ISS

NASA Image and Video Library

2012-09-28

ISS033-E-007920 (28 Sept. 2012) --- European Space Agency's "Edoardo Amaldi" Automated Transfer Vehicle-3 (ATV-3) begins its relative separation from the International Space Station during the Expedition 33 mission. The ATV-3 undocked from the aft port of the Zvezda Service Module at 5:44 p.m. (EDT) on Sept. 28, 2012. The ATV-3 is scheduled to deorbit on Oct. 2 for a fiery re-entry over the Pacific Ocean that will destroy the trash-filled spacecraft. Inside the ATV-3 is the Re-Entry Breakup Recorder that will record various data such as temperature, pressure and speed as the resupply craft burns up during its return to Earth. Experts will use that data to design safer and more predictable destructive re-entry techniques.

Recovery, Transportation and Acceptance to the Curation Facility of the Hayabusa Re-Entry Capsule

NASA Technical Reports Server (NTRS)

Abe, M.; Fujimura, A.; Yano, H.; Okamoto, C.; Okada, T.; Yada, T.; Ishibashi, Y.; Shirai, K.; Nakamura, T.; Noguchi, T.;

Development Of Metallic Thermal Protection System For The Expert Re-Entry Vehicle: Design Verification

NASA Astrophysics Data System (ADS)

Fatemi, Javad

2011-05-01

The thermal protection system of the EXPERT re-entry vehicle is subjected to accelerations, vibrations, acoustic and shock loads during launch and aero-heating loads and aerodynamic forces during re-entry. To fully understand the structural and thermomechanical performances of the TPS, heat transfer analysis, thermal stress analysis, and thermal buckling analysis must be performed. This requires complex three-dimensional thermal and structural models of the entire TPS including the insulation and sensors. Finite element (FE) methods are employed to assess the thermal and structural response of the TPS to the mechanical and aerothermal loads. The FE analyses results are used for the design verification and design improvement of the EXPERT thermal protection system.

Controlling the response to DNA damage by the APC/C-Cdh1.

PubMed

de Boer, H Rudolf; Guerrero Llobet, S; van Vugt, Marcel A T M

2016-03-01

Proper cell cycle progression is safeguarded by the oscillating activities of cyclin/cyclin-dependent kinase complexes. An important player in the regulation of mitotic cyclins is the anaphase-promoting complex/cyclosome (APC/C), a multi-subunit E3 ubiquitin ligase. Prior to entry into mitosis, the APC/C remains inactive, which allows the accumulation of mitotic regulators. APC/C activation requires binding to either the Cdc20 or Cdh1 adaptor protein, which sequentially bind the APC/C and facilitate targeting of multiple mitotic regulators for proteasomal destruction, including Securin and Cyclin B, to ensure proper chromosome segregation and mitotic exit. Emerging data have indicated that the APC/C, particularly in association with Cdh1, also functions prior to mitotic entry. Specifically, the APC/C-Cdh1 is activated in response to DNA damage in G2 phase cells. These observations are in line with in vitro and in vivo genetic studies, in which cells lacking Cdh1 expression display various defects, including impaired DNA repair and aberrant cell cycle checkpoints. In this review, we summarize the current literature on APC/C regulation in response to DNA damage, the functions of APC/C-Cdh1 activation upon DNA damage, and speculate how APC/C-Cdh1 can control cell fate in the context of persistent DNA damage.

9 CFR 93.421 - Special provisions.

Code of Federal Regulations, 2011 CFR

2011-01-01

... this chapter for entry into Canada: Provided, That all ruminants offered for re-entry upon examination... immediate export shall be inspected at the border port of entry and, when accompanied by an import permit obtained under § 93.404 of this part and all conditions therein are observed, shall be allowed entry into...

9 CFR 93.421 - Special provisions.

Code of Federal Regulations, 2013 CFR

2013-01-01

... this chapter for entry into Canada: Provided, That all ruminants offered for re-entry upon examination... immediate export shall be inspected at the border port of entry and, when accompanied by an import permit obtained under § 93.404 of this part and all conditions therein are observed, shall be allowed entry into...

9 CFR 93.421 - Special provisions.

Code of Federal Regulations, 2012 CFR

2012-01-01

... this chapter for entry into Canada: Provided, That all ruminants offered for re-entry upon examination... immediate export shall be inspected at the border port of entry and, when accompanied by an import permit obtained under § 93.404 of this part and all conditions therein are observed, shall be allowed entry into...

9 CFR 93.421 - Special provisions.

Code of Federal Regulations, 2014 CFR

2014-01-01

... this chapter for entry into Canada: Provided, That all ruminants offered for re-entry upon examination... immediate export shall be inspected at the border port of entry and, when accompanied by an import permit obtained under § 93.404 of this part and all conditions therein are observed, shall be allowed entry into...

9 CFR 93.421 - Special provisions.

Code of Federal Regulations, 2010 CFR

2010-01-01

... this chapter for entry into Canada: Provided, That all ruminants offered for re-entry upon examination... immediate export shall be inspected at the border port of entry and, when accompanied by an import permit obtained under § 93.404 of this part and all conditions therein are observed, shall be allowed entry into...

Heat Shock Protein B1-Deficient Mice Display Impaired Wound Healing

PubMed Central

McNamee, Kay; Przybycien, Paulina M.; Lu, Xin; Williams, Richard O.; Bou-Gharios, George; Saklatvala, Jeremy; Dean, Jonathan L. E.

2013-01-01

There is large literature describing in vitro experiments on heat shock protein (hsp)B1 but understanding of its function in vivo is limited to studies in mice overexpressing human hspB1 protein. Experiments in cells have shown that hspB1 has chaperone activity, a cytoprotective role, regulates inflammatory gene expression, and drives cell proliferation. To investigate the function of the protein in vivo we generated hspB1-deficient mice. HspB1-deficient fibroblasts display increased expression of the pro-inflammatory cytokine, interleukin-6, compared to wild-type cells, but reduced proliferation. HspB1-deficient fibroblasts exhibit reduced entry into S phase and increased expression of cyclin-dependent kinase inhibitors p27kip1 and p21waf1. The expression of hspB1 protein and mRNA is also controlled by the cell cycle. To investigate the physiological function of hspB1 in regulating inflammation and cell proliferation we used an excisional cutaneous wound healing model. There was a significant impairment in the rate of healing of wounds in hspB1-deficient mice, characterised by reduced re-epithelialisation and collagen deposition but also increased inflammation. HspB1 deficiency augments neutrophil infiltration in wounds, driven by increased chemokine (C-X-C motif) ligand 1 expression. This appears to be a general mechanism as similar results were obtained in the air-pouch and peritonitis models of acute inflammation. PMID:24143227

Assessment of Aerothermodynamic Flight Prediction Tools through Ground and Flight Experimentation (Evaluation des outils aerothermodynamiques de prediction de vol par l’experimentation au sol et en vol)

DTIC Science & Technology

2011-11-01

6.5 Conclusions 6-30 6.6 Acknowledgments 6-31 6.7 References 6-31 Chapter 7 – Experimental Investigation of the Supersonic Wake of a Re-entry 7-1... Noise on the Axial Location of 4-4 Transition for the HIFiRE-1 Cone at Zero Angle of Attack and Mach 6 Figure 4-2 Correlations for Transition...Without Sting) Mach 2 AoA 19 Symmetry Plane Computed 6-14 with LORE: Effect of Sting/ Blade vs. No Sting/ Blade on 8M Cells Mesh Figure 6-12 Mach 2

Flow Redistribution Between Legs and Brain During STS 93 Re-Entry and Landing

NASA Technical Reports Server (NTRS)

Arbeille, P.; Meck, J.; Porcher, M.; Benavides, E.; Martin, D. S.; South, D. A.; Ribeiro, C.; Westover, A.

2003-01-01

The objective was to quantify bit by bit the arterial hemodynamic response to the successive acceleration induced fluid shifts during re-entry and landing. Method: The astronaut instrumented himself with a flat Doppler probe fixed on the skin, a blood pressure arm cuff, and 3 ECG electrodes. The ICMS (integrated cardiovascular monitoring system, 15x15x25 cu cm, battery powered) designed to monitor Blood pressure, ECG, cerebral and femoral flows was fixed below the astronaut sit in the middeck. Recordings started 5 minutes before de-orbiting (TIG) and stopped 5 min after wheels stop. Results. During re-entry blood pressure increased by 20% at TIG, and then by 25 to 30% during the highest Gz accelerations (approx 1 S g ) . The cerebral flow remained decreased by 10 to 15% below inflight value all during the Entry and landing phases. Conversely the femoral flow increased at TIG and entry ( + l0 to 20%), recovered at 0.lg, and then decreased in proportion with the Gz acceleration (-10% to -40% from 0.5g to 1.5g). The reduction in Femoral flow was associated with an opposite variation in lower limb vascular resistance. Consequently the cerebral flow/femoral flow ratio decreased at TIG and entry (-20%), and then increased according to the Gz acceleration level ( + l0 to +40% from 0.5 to 1.5g). Conclusion: During orthostatic tests (Stand LBNP tests) the cerebral to femoral flow ratio allowed to quantify the efficiency of the flow redistribution between these 2 areas and predicted orthostatic intolerance. In the present case the astronaut was found orthostatically tolerant at postflight tilt tests, but we suggest that during re-entry this parameter could predict the occurrence of syncope in severely disadapted astronauts.

Automated Re-Entry System using FNPEG

NASA Technical Reports Server (NTRS)

Johnson, Wyatt R.; Lu, Ping; Stachowiak, Susan J.

2017-01-01

This paper discusses the implementation and simulated performance of the FNPEG (Fully Numerical Predictor-corrector Entry Guidance) algorithm into GNC FSW (Guidance, Navigation, and Control Flight Software) for use in an autonomous re-entry vehicle. A few modifications to FNPEG are discussed that result in computational savings -- a change to the state propagator, and a modification to cross-range lateral logic. Finally, some Monte Carlo results are presented using a representative vehicle in both a high-fidelity 6-DOF (degree-of-freedom) sim as well as in a 3-DOF sim for independent validation.

Wiring Zinc in Three Dimensions Re-writes Battery Performance - Dendrite-Free Cycling

DTIC Science & Technology

2014-01-01

surfaces throughout the electrode structure (Fig. 5D–I). The positive Zn@ZnO sponge exhibits a compact morphology uniformly distributed throughout (Fig...monolithic, three-dimensional (3D) aperiodic architecture. Utilization approaches 90% (728 mA h gZn 1) when the zinc “ sponge ” is used as the anode in...a primary (single-use) zinc–air cell. To probe rechargeability of the 3D Zn sponge , we cycled Zn–vs.–Zn symmetric cells and Ag–Zn full cells under

Electron Microscopy of Ebola Virus-Infected Cells.

PubMed

Noda, Takeshi

2017-01-01

Ebola virus (EBOV) replicates in host cells, where both viral and cellular components show morphological changes during the process of viral replication from entry to budding. These steps in the replication cycle can be studied using electron microscopy (EM), including transmission electron microscopy (TEM) and scanning electron microscopy (SEM), which is one of the most useful methods for visualizing EBOV particles and EBOV-infected cells at the ultrastructural level. This chapter describes conventional methods for EM sample preparation of cultured cells infected with EBOV.

PLC-γ1 Signaling Plays a Subtype-Specific Role in Postbinding Cell Entry of Influenza A Virus

PubMed Central

Zhu, Liqian; Ly, Hinh

2014-01-01

Host signaling pathways and cellular proteins play important roles in the influenza viral life cycle and can serve as antiviral targets. In this study, we report the engagement of host phosphoinositide-specific phospholipase γ1 (PLC-γ1) in mediating cell entry of influenza virus H1N1 but not H3N2 subtype. Both PLC-γ1-specific inhibitor and short hairpin RNA (shRNA) strongly suppress the replication of H1N1 but not H3N2 viruses in cell culture, suggesting that PLC-γ1 plays an important subtype-specific role in the influenza viral life cycle. Further analyses demonstrate that PLC-γ1 activation is required for viral postbinding cell entry. In addition, H1N1, but not H3N2, infection leads to the phosphorylation of PLC-γ1 at Ser 1248 immediately after infection and independent of viral replication. We have further shown that H1N1-induced PLC-γ1 activation is downstream of epidermal growth factor receptor (EGFR) signaling. Interestingly, both H1N1 and H3N2 infections activate EGFR, but only H1N1 infection leads to PLC-γ1 activation. Taking our findings together, we have identified for the first time the subtype-specific interplay of host PLC-γ1 signaling and H1N1 virus that is critical for viral uptake early in the infection. Our study provides novel insights into how virus interacts with the cellular signaling network by demonstrating that viral determinants can regulate how the host signaling pathways function in virally infected cells. PMID:24155396

Cell cycle-dependent regulation of Greatwall kinase by protein phosphatase 1 and regulatory subunit 3B.

PubMed

Ren, Dapeng; Fisher, Laura A; Zhao, Jing; Wang, Ling; Williams, Byron C; Goldberg, Michael L; Peng, Aimin

2017-06-16

Greatwall (Gwl) kinase plays an essential role in the regulation of mitotic entry and progression. Mitotic activation of Gwl requires both cyclin-dependent kinase 1 (CDK1)-dependent phosphorylation and its autophosphorylation at an evolutionarily conserved serine residue near the carboxyl terminus (Ser-883 in Xenopus ). In this study we show that Gwl associates with protein phosphatase 1 (PP1), particularly PP1γ, which mediates the dephosphorylation of Gwl Ser-883. Consistent with the mitotic activation of Gwl, its association with PP1 is disrupted in mitotic cells and egg extracts. During mitotic exit, PP1-dependent dephosphorylation of Gwl Ser-883 occurs prior to dephosphorylation of other mitotic substrates; replacing endogenous Gwl with a phosphomimetic S883E mutant blocks mitotic exit. Moreover, we identified PP1 regulatory subunit 3B (PPP1R3B) as a targeting subunit that can direct PP1 activity toward Gwl. PPP1R3B bridges PP1 and Gwl association and promotes Gwl Ser-883 dephosphorylation. Consistent with the cell cycle-dependent association of Gwl and PP1, Gwl and PPP1R3B dissociate in M phase. Interestingly, up-regulation of PPP1R3B facilitates mitotic exit and blocks mitotic entry. Thus, our study suggests PPP1R3B as a new cell cycle regulator that functions by governing Gwl dephosphorylation. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

The risk of stanford type-A aortic dissection with different tear size and location: a numerical study.

PubMed

Shi, Yue; Zhu, Minjia; Chang, Yu; Qiao, Huanyu; Liu, Yongmin

2016-12-28

This study is to investigate the influence of hemodynamics on Stanford type-A aortic dissection with different tear size and location, to provide some support for the relationships between the risks (rupture, reverse tearing and further tearing) and tear size and location for clinical treatment. Four numerical models of Stanford type-A aortic dissection were established, with different size and location of the tears. The ratio of the area between the entry and re-entry tears(RA) is various within the model; while, the size and the location of the re-entry in the distal descending aorta are fixed. In model A11 and A21, the entry tears are located near the ascending aorta. The RA in these models are 1 and 2, respectively; In the model B11 and B21, the entry tears are located near the proximal descending aorta and the RA in these models are again assigned to 1 and 2, respectively. Then hemodynamics in these models was solved with numerically and the flow patterns and loading distributions were investigated. The flow velocity of the true lumen in model A21, B21 is lower than that in A11, B11, respectively; the time-averaged wall shear stress (TAWSS) of the false lumen in model A21 and B21 is higher, and for ascending aorta false lumen, A11, A21 are higher than B11, B21, respectively. False lumen intimal wall pressure of A11, A21 are always higher than the true lumen ones. The variation of the RA can significantly affect the dynamics of blood within the aortic dissection. When the entry tear size is larger than the re-entry tear ones, the false lumen, proximal descending aorta and the wall near re-entry tear are prone to cracking. Entry tear location can significantly alter the hemodynamics of aortic dissection as well. When entry tear location is closer to proximal ascending aorta, false lumen continues to expand and compress the true lumen resulting in the true lumen reduction. For proximal ascending aorta, high pressure in false lumen predicts a higher risk of reverse tear.

Investigations into the Properties, Conditions, and Effects of the Ionosphere.

DTIC Science & Technology

1988-01-15

Innovative Approaches to Direct Measurement of N e 28 ]- J FEASIBILITY OF RADIO BLACKOUT MITIGATION IN THE BRAKING PHASE 28 OF AOTV OPERATIONS 1. Brief...Cell could be tested under simulated flight conditions in the SAIC plasma laboratory facility. AJ. FEASIBILITY OF RADIO BLACKOUT MITIGATION IN THE...enable calculation of chemical modification techniques ,. on phenomena of radio blackout during re-entry of orbiting spacecraft . * ,1. Brief

Electrode behavior RE-visited: Monitoring potential windows, capacity loss, and impedance changes in Li 1.03 (Ni 0.5Co 0.2Mn 0.3) 0.97O 2/silicon-graphite full cells

DOE PAGES

Klett, Matilda; Gilbert, James A.; Trask, Stephen E.; ...

2016-03-04

Here, the capacity and power performance of lithium-ion battery cells evolve over time. The mechanisms leading to these changes can often be identified through knowledge of electrode potentials, which contain information about electrochemical processes at the electrode-electrolyte interfaces. In this study we monitor electrode potentials within full cells containing a Li 1.03(Ni 0.5Co 0.2Mn 0.3) 0.97O 2–based (NCM523) positive electrode, a silicon-graphite negative electrode, and an LiPF6-bearing electrolyte, with and without fluoroethylene carbonate (FEC) or vinylene carbonate (VC) additives. The electrode potentials are monitored with a Li-metal reference electrode (RE) positioned besides the electrode stack; changes in these potentials aremore » used to examine electrode state-of-charge (SOC) shifts, material utilization, and loss of electrochemically active material. Electrode impedances are obtained with a Li xSn RE located within the stack; the data display the effect of cell voltage and electrode SOC changes on the measured values after formation cycling and after aging. Our measurements confirm the beneficial effect of FEC and VC electrolyte additives in reducing full cell capacity loss and impedance rise after cycling in a 3.0–4.2 V range. Comparisons with data from a full cell containing a graphite-based negative highlight the consequences of including silicon in the electrode. Our observations on electrode potentials, capacity, and impedance changes on cycling are crucial to designing long-lasting, silicon-bearing, lithium-ion cells.« less

Spatio-temporal re-organization of replication foci accompanies replication domain consolidation during human pluripotent stem cell lineage specification

PubMed Central

Wilson, Korey A.; Elefanty, Andrew G.; Stanley, Edouard G.; Gilbert, David M.

2016-01-01

ABSTRACT Lineage specification of both mouse and human pluripotent stem cells (PSCs) is accompanied by spatial consolidation of chromosome domains and temporal consolidation of their replication timing. Replication timing and chromatin organization are both established during G1 phase at the timing decision point (TDP). Here, we have developed live cell imaging tools to track spatio-temporal replication domain consolidation during differentiation. First, we demonstrate that the fluorescence ubiquitination cell cycle indicator (Fucci) system is incapable of demarcating G1/S or G2/M cell cycle transitions. Instead, we employ a combination of fluorescent PCNA to monitor S phase progression, cytokinesis to demarcate mitosis, and fluorescent nucleotides to label early and late replication foci and track their 3D organization into sub-nuclear chromatin compartments throughout all cell cycle transitions. We find that, as human PSCs differentiate, the length of S phase devoted to replication of spatially clustered replication foci increases, coincident with global compartmentalization of domains into temporally clustered blocks of chromatin. Importantly, re-localization and anchorage of domains was completed prior to the onset of S phase, even in the context of an abbreviated PSC G1 phase. This approach can also be employed to investigate cell fate transitions in single PSCs, which could be seen to differentiate preferentially from G1 phase. Together, our results establish real-time, live-cell imaging methods for tracking cell cycle transitions during human PSC differentiation that can be applied to study chromosome domain consolidation and other aspects of lineage specification. PMID:27433885

Does Offender Gambling on the inside Continue on the outside? Insights from Correctional Professionals on Gambling and Re-Entry

ERIC Educational Resources Information Center

Williams, D. J.; Walker, Gordon J.

2009-01-01

This study brings to light a neglected topic of particular importance--offender gambling issues within the context of re-entry into the community. Fifteen correctional professionals from Nevada (high gambling availability) and Utah (no legalized gambling) participated in semi-structured interviews to provide insights into how gambling may impact…

STD 105: Process Groups as an Instructional Medium for Re-entry Women at Paul D. Camp Community College.

ERIC Educational Resources Information Center

Creamer, Elizabeth; Duggin, Molly; Kidd, Ronald

1999-01-01

An effective team-based, group-oriented personal development from a woman's perspective course explores the effects of several issues on the re-entry woman: the role that society and culture play in influencing women's vocational choices; women's roles; the economic necessity of work; stress; and relationships. A team-based approach provides a…

8 CFR 212.2 - Consent to reapply for admission after deportation, removal or departure at Government expense.

Code of Federal Regulations, 2010 CFR

2010-01-01

... United States for the time period required for re-entry after deportation or removal. The examining... years from the deportation date before he or she is eligible to re-enter the United States. Any alien... permission to reapply for entry in conjunction with his or her application for adjustment of status. This...

An Exploration of Factors Reducing Recidivism Rates of Formerly Incarcerated Youth with Disabilities Participating in a Re-Entry Intervention

ERIC Educational Resources Information Center

Unruh, Deanne K.; Gau, Jeff M.; Waintrup, Miriam G.

2009-01-01

Juvenile offenders are costly to our society in terms of the monetary and social expenditures from the legal system, victims' person costs, and incarceration. The re-entry and community reintegration outcomes for formerly incarcerated youth with a disabling condition are bleak compared to peers without disabilities. In this study, we examined the…

8 CFR 212.2 - Consent to reapply for admission after deportation, removal or departure at Government expense.

Code of Federal Regulations, 2011 CFR

2011-01-01

... United States for the time period required for re-entry after deportation or removal. The examining... years from the deportation date before he or she is eligible to re-enter the United States. Any alien... permission to reapply for entry in conjunction with his or her application for adjustment of status. This...

8 CFR 212.2 - Consent to reapply for admission after deportation, removal or departure at Government expense.

Code of Federal Regulations, 2013 CFR

2013-01-01

... United States for the time period required for re-entry after deportation or removal. The examining... years from the deportation date before he or she is eligible to re-enter the United States. Any alien... of the Act and part 245 of this chapter must request permission to reapply for entry in conjunction...

8 CFR 212.2 - Consent to reapply for admission after deportation, removal or departure at Government expense.

Code of Federal Regulations, 2012 CFR

2012-01-01

... United States for the time period required for re-entry after deportation or removal. The examining... years from the deportation date before he or she is eligible to re-enter the United States. Any alien... of the Act and part 245 of this chapter must request permission to reapply for entry in conjunction...

8 CFR 212.2 - Consent to reapply for admission after deportation, removal or departure at Government expense.

Code of Federal Regulations, 2014 CFR

2014-01-01

... United States for the time period required for re-entry after deportation or removal. The examining... years from the deportation date before he or she is eligible to re-enter the United States. Any alien... of the Act and part 245 of this chapter must request permission to reapply for entry in conjunction...

Evaluation of a school re-entry nursing intervention for children with cancer.

PubMed

McCarthy, A M; Williams, J; Plumer, C

1998-07-01

A retrospective qualitative design was used to identify and compare the concerns, parents, teachers, and children have regarding school re-entry after a cancer diagnosis and to describe the impact of a school re-entry program on parents', teachers', and children's concerns. Audiotaped, semistructured interviews were obtained from a convenience sample of 10 children with cancer (ages 5 to 13 years), 10 mothers, and nine teachers. All participants were positive about the school re-entry nursing intervention, which is described. Results of content analyses indicate that before the intervention, mothers were concerned about their child's safety and peer teasing; teachers were concerned about their own knowledge and peers' adjustment, and children were concerned with keeping up with school activities. After the intervention, mothers were less concerned about peer teasing but continued to be worried about their child's safety in the school setting and began to have concerns about academic progress and physical stamina; teachers reported increased concerns for the child's safety and academic progress, and a desire to return to normal routines in the classroom: and the children continued to have concerns with maintaining academic and/physical progress. Clinical and research implications are discussed.

The ESA/NASA Multi-Aircraft ATV-1 Re-Entry Campaign: Analysis of Airborne Intensified Video Observations from the NASA/JSC Experiment

NASA Technical Reports Server (NTRS)

Barker, Ed; Maley, Paul; Mulrooney, Mark; Beaulieu, Kevin

2009-01-01

In September 2008, a joint ESA/NASA multi-instrument airborne observing campaign was conducted over the Southern Pacific ocean. The objective was the acquisition of data to support detailed atmospheric re-entry analysis for the first flight of the European Automated Transfer Vehicle (ATV)-1. Skilled observers were deployed aboard two aircraft which were flown at 12.8 km altitude within visible range of the ATV-1 re-entry zone. The observers operated a suite of instruments with low-light-level detection sensitivity including still cameras, high speed and 30 fps video cameras, and spectrographs. The collected data has provided valuable information regarding the dynamic time evolution of the ATV-1 re-entry fragmentation. Specifically, the data has satisfied the primary mission objective of recording the explosion of ATV-1's primary fuel tank and thereby validating predictions regarding the tanks demise and the altitude of its occurrence. Furthermore, the data contains the brightness and trajectories of several hundred ATV-1 fragments. It is the analysis of these properties, as recorded by the particular instrument set sponsored by NASA/Johnson Space Center, which we present here.

Proteomic analysis of the response to cell cycle arrests in human myeloid leukemia cells

PubMed Central

Ly, Tony; Endo, Aki; Lamond, Angus I

2015-01-01

Abstract Previously, we analyzed protein abundance changes across a ‘minimally perturbed’ cell cycle by using centrifugal elutriation to differentially enrich distinct cell cycle phases in human NB4 cells (Ly et al., 2014). In this study, we compare data from elutriated cells with NB4 cells arrested at comparable phases using serum starvation, hydroxyurea, or RO-3306. While elutriated and arrested cells have similar patterns of DNA content and cyclin expression, a large fraction of the proteome changes detected in arrested cells are found to reflect arrest-specific responses (i.e., starvation, DNA damage, CDK1 inhibition), rather than physiological cell cycle regulation. For example, we show most cells arrested in G2 by CDK1 inhibition express abnormally high levels of replication and origin licensing factors and are likely poised for genome re-replication. The protein data are available in the Encyclopedia of Proteome Dynamics (http://www.peptracker.com/epd/), an online, searchable resource. DOI: http://dx.doi.org/10.7554/eLife.04534.001 PMID:25555159

Intrinsic and extrinsic mechanisms regulating satellite cell function

PubMed Central

Dumont, Nicolas A.; Wang, Yu Xin; Rudnicki, Michael A.

2015-01-01

Muscle stem cells, termed satellite cells, are crucial for skeletal muscle growth and regeneration. In healthy adult muscle, satellite cells are quiescent but poised for activation. During muscle regeneration, activated satellite cells transiently re-enter the cell cycle to proliferate and subsequently exit the cell cycle to differentiate or self-renew. Recent studies have demonstrated that satellite cells are heterogeneous and that subpopulations of satellite stem cells are able to perform asymmetric divisions to generate myogenic progenitors or symmetric divisions to expand the satellite cell pool. Thus, a complex balance between extrinsic cues and intrinsic regulatory mechanisms is needed to tightly control satellite cell cycle progression and cell fate determination. Defects in satellite cell regulation or in their niche, as observed in degenerative conditions such as aging, can impair muscle regeneration. Here, we review recent discoveries of the intrinsic and extrinsic factors that regulate satellite cell behaviour in regenerating and degenerating muscles. PMID:25922523

Molecular identification of PpHDAC1, the first histone deacetylase fron the slime mold Physarum polycephalum.

PubMed

Brandtner, Eva-Maria; Lechner, Thomas; Loidl, Peter; Lusser, Alexandra

2002-01-01

The dynamic state of post-translational acetylation of eukaryotic histones is maintained by histone acetyltransferases (HATs) and histone deacetylases (HDACs). HATs and HDACs have been shown to be components of various regulatory protein complexes in the cell. Their enzymatic activities, intracellular localization and substrate specificities are regulated in a complex, cell cycle related manner. In the myxomycete Physarum polycephalum multiple HATs and HDACs can be distinguished in biochemical terms and they exhibit dynamic activity patterns depending on the cell cycle stage. Here we report on the cloning of the first P. polycephalum HDAC (PpHDAC1) related to the S. cerevisiae Rpd3 protein. The expression pattern of PpHDAC1 mRNA was analysed at different time points of the cell cycle and found to be largely constant. Treatment of macroplasmodia with the HDAC inhibitor trichostatin A at several cell cycle stages resulted in a significant delay in entry into mitosis of treated versus untreated plasmodia. No effect of TSA treatment could be observed on PpHDAC1 expression itself.

9 CFR 93.318 - Special provisions.

Code of Federal Regulations, 2010 CFR

2010-01-01

... for re-entry upon examination by the veterinary inspector at the U.S. port of entry, are found by the... be inspected at the border port of entry and, when accompanied by an import permit obtained under § 93.304 of this part and all conditions therein are observed, shall be allowed entry into the United...

9 CFR 93.318 - Special provisions.

Code of Federal Regulations, 2013 CFR

2013-01-01

... for re-entry upon examination by the veterinary inspector at the U.S. port of entry, are found by the... be inspected at the border port of entry and, when accompanied by an import permit obtained under § 93.304 of this part and all conditions therein are observed, shall be allowed entry into the United...

9 CFR 93.519 - Special provisions.

Code of Federal Regulations, 2014 CFR

2014-01-01

... offered for re-entry upon examination by the veterinary inspector at the U.S. port of entry, are found by... inspected at the border port of entry and, when accompanied by an import permit obtained under § 93.504 of this part and all conditions therein are observed, shall be allowed entry into the United States and...

9 CFR 93.519 - Special provisions.

Code of Federal Regulations, 2012 CFR

2012-01-01

... offered for re-entry upon examination by the veterinary inspector at the U.S. port of entry, are found by... inspected at the border port of entry and, when accompanied by an import permit obtained under § 93.504 of this part and all conditions therein are observed, shall be allowed entry into the United States and...

9 CFR 93.318 - Special provisions.

Code of Federal Regulations, 2011 CFR

2011-01-01

... for re-entry upon examination by the veterinary inspector at the U.S. port of entry, are found by the... be inspected at the border port of entry and, when accompanied by an import permit obtained under § 93.304 of this part and all conditions therein are observed, shall be allowed entry into the United...

9 CFR 93.519 - Special provisions.

Code of Federal Regulations, 2013 CFR

2013-01-01

... offered for re-entry upon examination by the veterinary inspector at the U.S. port of entry, are found by... inspected at the border port of entry and, when accompanied by an import permit obtained under § 93.504 of this part and all conditions therein are observed, shall be allowed entry into the United States and...

9 CFR 93.318 - Special provisions.

Code of Federal Regulations, 2012 CFR

2012-01-01

... for re-entry upon examination by the veterinary inspector at the U.S. port of entry, are found by the... be inspected at the border port of entry and, when accompanied by an import permit obtained under § 93.304 of this part and all conditions therein are observed, shall be allowed entry into the United...

9 CFR 93.519 - Special provisions.

Code of Federal Regulations, 2011 CFR

2011-01-01

... offered for re-entry upon examination by the veterinary inspector at the U.S. port of entry, are found by... inspected at the border port of entry and, when accompanied by an import permit obtained under § 93.504 of this part and all conditions therein are observed, shall be allowed entry into the United States and...

9 CFR 93.519 - Special provisions.

Code of Federal Regulations, 2010 CFR

2010-01-01

... offered for re-entry upon examination by the veterinary inspector at the U.S. port of entry, are found by... inspected at the border port of entry and, when accompanied by an import permit obtained under § 93.504 of this part and all conditions therein are observed, shall be allowed entry into the United States and...

9 CFR 93.318 - Special provisions.

Code of Federal Regulations, 2014 CFR

2014-01-01

... for re-entry upon examination by the veterinary inspector at the U.S. port of entry, are found by the... be inspected at the border port of entry and, when accompanied by an import permit obtained under § 93.304 of this part and all conditions therein are observed, shall be allowed entry into the United...

Incarcerated women's relationship-based strategies to avoid drug use after community re-entry.

PubMed

Snell-Rood, Claire; Staton-Tindall, Michele; Victor, Grant

2016-10-01

While recent research has stressed the supportive role that family and friends play for incarcerated persons as they re-enter the community, drug-using incarcerated women re-entering the community often have to rely on family, community, and intimate relationships that have played a role in their substance abuse and criminalization. In this study the authors conducted qualitative analysis of clinical sessions with rural, drug-using women (N = 20) in a larger prison-based HIV risk reduction intervention in Kentucky during 2012-2014 to examine incarcerated women's perceptions of the role of their family, community, and intimate relationships in their plans to decrease their substance abuse upon community re-entry. Women stressed the obstacles to receiving support in many of their family and drug-using relationships after community re-entry. Nonetheless, they asserted that changes in their relationships could support their desires to end their substance abuse by setting limits on and using their positive relationships, particularly with their children, to motivate them to change. Interventions to promote incarcerated women's health behavior changes-including substance abuse-must acknowledge the complex social environments in which they live.

Cyclin B in mouse oocytes and embryos: importance for human reproduction and aneuploidy.

PubMed

Polański, Zbigniew; Homer, Hayden; Kubiak, Jacek Z

2012-01-01

Oocyte maturation and early embryo development require precise coordination between cell cycle progression and the developmental programme. Cyclin B plays a major role in this process: its accumulation and degradation is critical for driving the cell cycle through activation and inactivation of the major cell cycle kinase, CDK1. CDK1 activation is required for M-phase entry whereas its inactivation leads to exit from M-phase. The tempo of oocyte meiotic and embryonic mitotic divisions is set by the rate of cyclin B accumulation and the timing of its destruction. By controlling when cyclin B destruction is triggered and by co-ordinating this with the completion of chromosome alignment, the spindle assembly checkpoint (SAC) is a critical quality control system important for averting aneuploidy and for building in the flexibility required to better integrate cell cycle progression with development. In this review we focus on cyclin B metabolism in mouse oocytes and embryos and illustrate how the cell cycle-powered clock (in fact cyclin B-powered clock) controls oocyte maturation and early embryo development, thereby providing important insight into human reproduction and potential causes of Down syndrome.

Suppression of Myc oncogenic activity by ribosomal protein haploinsufficiency

PubMed Central

Barna, Maria; Pusic, Aya; Zollo, Ornella; Costa, Maria; Kondrashov, Nadya; Rego, Eduardo; Rao, Pulivarthi H; Ruggero, Davide

2008-01-01

The Myc oncogene regulates the expression of multiple components of the protein synthetic machinery, including ribosomal proteins, initiation factors of translation, Pol III, and rDNA1,2. An outstanding question is whether and how increasing the cellular protein synthesis capacity can affect the multi-step process leading to cancer. We utilized ribosomal protein heterozygote mice as a genetic tool to restore increased protein synthesis in Eμ–Myc/+ transgenic mice to normal levels and show that in this context Myc's oncogenic potential is suppressed. Our findings demonstrate that the ability of Myc to increase protein synthesis directly augments cell size and is sufficient to accelerate cell cycle progression independently of known cell cycle targets transcriptionally regulated by Myc. In addition, when protein synthesis is restored to normal levels, Myc overexpressing precancerous cells are more efficiently eliminated by programmed cell death. Our findings reveal a novel paradigm that links increases in general protein synthesis rates downstream of an oncogenic signal to a specific molecular impairment in the modality of translation initiation employed to regulate the expression of selective mRNAs. We show that an aberrant increase in cap-dependent translation downstream Myc hyperactivation specifically impairs the translational switch to internal ribosomal entry site (IRES)-dependent translation required for accurate mitotic progression. Failure of this translational switch results in reduced mitotic-specific expression of the endogenous IRES-dependent form of Cdk11 (p58-PITSLRE)3-5, which leads to cytokinesis defects and is associated with increased centrosome numbers and genome instability in Eμ–Myc/+ mice. When accurate translational control is re-established in Eμ–Myc/+ mice, genome instability is suppressed. Our findings reveal how perturbations in translational control provide a highly specific outcome on gene expression, genome stability, and cancer initiation that have important implications for understanding the molecular mechanism of cancer formation at the post-genomic level. PMID:19011615

Protein tyrosine phosphatase, PTP1B, expression and activity in rat corneal endothelial cells

PubMed Central

Harris, Deshea L.

2007-01-01

Purpose The current studies were conducted to determine whether the protein tyrosine phosphatase, PTP1B, plays a role in regulating epidermal growth factor receptor (EGFR) Tyr992 phosphorylation and cell cycle entry in rat corneal endothelial cells. Methods Corneas were obtained from male Sprague-Dawley rats. PTP1B mRNA and protein expression were compared in confluent and subconfluent cells by RT-PCR and western blots. Immunocytochemistry was used to determine the subcellular localization of both PTP1B and EGFR following epidermal growth factor (EGF) stimulation. Western blots were used to analyze the time-dependent effect of EGF on phosphorylation of EGFR Tyr992 plus or minus CinnGEL 2Me, an inhibitor of PTP1B activity. The effect of PTP1B inhibition on cell cycle entry was determined by calculating the percent of Ki67-positive cells following EGF treatment. Results PTP1B mRNA expression was similar in confluent and subconfluent cells, but PTP1B protein was expressed at 3 fold higher levels in subconfluent cells. Positive staining for PTP1B was localized in vesicular structures below the plasma membrane. EGFR staining was located at cell-cell borders in untreated endothelium, but was mainly cytoplasmic by 15 min after EGF treatment. In control cultures, phosphorylation of EGFR Tyr992 peaked by 5 min following EGF stimulation and rapidly decreased to basal levels by 30 min. In cultures pretreated with CinnGEL 2Me, Tyr992 phosphorylation peaked 2 min following EGF addition and was consistently sustained at a higher level than controls until 60 min after treatment. By 18 h following EGF treatment, cultures pretreated with CinnGEL 2Me exhibited a 1.7 fold increase in the number of Ki67-positive cells compared with control cultures. Conclusions Comparison of PTP1B mRNA and protein levels indicates that PTP1B expression is regulated mainly at the protein level and is higher in subconfluent cells. PTP1B was located in vesicles below the plasma membrane. The fact that EGFR is internalized in response to EGF stimulation suggests that it could interact with and be regulated by PTP1B. The ability of PTP1B inhibitor to sustain EGFR Tyr992 phosphorylation and increase the number of Ki67-positive cells indicates that PTP1B plays a role in the negative regulation of EGF-induced signaling and helps suppress cell cycle entry. PMID:17563729

The Notch pathway regulates the Second Mitotic Wave cell cycle independently of bHLH proteins.

PubMed

Bhattacharya, Abhishek; Li, Ke; Quiquand, Manon; Rimesso, Gerard; Baker, Nicholas E

2017-11-15

Notch regulates both neurogenesis and cell cycle activity to coordinate precursor cell generation in the differentiating Drosophila eye. Mosaic analysis with mitotic clones mutant for Notch components was used to identify the pathway of Notch signaling that regulates the cell cycle in the Second Mitotic Wave. Although S phase entry depends on Notch signaling and on the transcription factor Su(H), the transcriptional co-activator Mam and the bHLH repressor genes of the E(spl)-Complex were not essential, although these are Su(H) coactivators and targets during the regulation of neurogenesis. The Second Mitotic Wave showed little dependence on ubiquitin ligases neuralized or mindbomb, and although the ligand Delta is required non-autonomously, partial cell cycle activity occurred in the absence of known Notch ligands. We found that myc was not essential for the Second Mitotic Wave. The Second Mitotic Wave did not require the HLH protein Extra macrochaetae, and the bHLH protein Daughterless was required only cell-nonautonomously. Similar cell cycle phenotypes for Daughterless and Atonal were consistent with requirement for neuronal differentiation to stimulate Delta expression, affecting Notch activity in the Second Mitotic Wave indirectly. Therefore Notch signaling acts to regulate the Second Mitotic Wave without activating bHLH gene targets. Copyright © 2017 Elsevier Inc. All rights reserved.

In vitro comparison of the antiproliferative effects of rhenium-186 and rhenium-188 on human aortic endothelial cells.

PubMed

Sauter, Alexander; Arthasana, Daniel; Dittmann, Helmut; Pritzkow, Maren; Wiesinger, Benjamin; Schmehl, Joerg; Brechtel, Klaus; Bantleon, Rüdiger; Claussen, Claus; Kehlbach, Rainer

2011-08-01

Rhenium-186 ((186)Re) and rhenium-188 ((188)Re) are promising radionuclides for the inhibition of restenosis after percutaneous transluminal angioplasty or other vascular interventions. Until now the maximal dose tolerance of endothelial cells has not been clearly known. To characterize the effects of local irradiation treatment, human aortic endothelial cells (ECs) were incubated with different doses of (186)Re and (188)Re. Two days after plating, ECs received treatment for a period of 5 days. The total radiation doses applied were 1, 4, 8, 16, and 32 Gy. On days 1, 3, 5, 7, and 12 after initial rhenium incubation, cell growth, clonogenic activity, cell-cycle distribution, and cytoskeletal architecture were evaluated. From the first day on, a dose-dependent growth inhibition was observed. Cumulative doses of ≥32 Gy caused a weak colony formation and significant alterations in the cytoskeletal architecture. An increased fraction of cells in G2/M phase was seen for cumulative radiation doses of ≥16 Gy. Interestingly, there were no significant differences between (186)Re and (188)Re. Even for low dose rates of β particles a dose-dependent proliferation inhibition of ECs is seen. Doses beyond 32 Gy alter the cytoskeletal architecture with possibly endothelial dysfunction and late thrombosis.

Etoposide Induces Nuclear Re-Localisation of AID

PubMed Central

Lambert, Laurens J.; Walker, Simon; Feltham, Jack; Lee, Heather J.; Reik, Wolf; Houseley, Jonathan

2013-01-01

During B cell activation, the DNA lesions that initiate somatic hypermutation and class switch recombination are introduced by activation-induced cytidine deaminase (AID). AID is a highly mutagenic protein that is maintained in the cytoplasm at steady state, however AID is shuttled across the nuclear membrane and the protein transiently present in the nucleus appears sufficient for targeted alteration of immunoglobulin loci. AID has been implicated in epigenetic reprogramming in primordial germ cells and cell fusions and in induced pluripotent stem cells (iPS cells), however AID expression in non-B cells is very low. We hypothesised that epigenetic reprogramming would require a pathway that instigates prolonged nuclear residence of AID. Here we show that AID is completely re-localised to the nucleus during drug withdrawal following etoposide treatment, in the period in which double strand breaks (DSBs) are repaired. Re-localisation occurs 2-6 hours after etoposide treatment, and AID remains in the nucleus for 10 or more hours, during which time cells remain live and motile. Re-localisation is cell-cycle dependent and is only observed in G2. Analysis of DSB dynamics shows that AID is re-localised in response to etoposide treatment, however re-localisation occurs substantially after DSB formation and the levels of re-localisation do not correlate with γH2AX levels. We conclude that DSB formation initiates a slow-acting pathway which allows stable long-term nuclear localisation of AID, and that such a pathway may enable AID-induced DNA demethylation during epigenetic reprogramming. PMID:24324754

Identification of Ellagic Acid from Plant Rhodiola rosea L. as an Anti-Ebola Virus Entry Inhibitor.

PubMed

Cui, Qinghua; Du, Ruikun; Anantpadma, Manu; Schafer, Adam; Hou, Lin; Tian, Jingzhen; Davey, Robert A; Cheng, Han; Rong, Lijun

2018-03-27

The recent 2014-2016 West African Ebola virus epidemic underscores the need for the development of novel anti-Ebola therapeutics, due to the high mortality rates of Ebola virus infections and the lack of FDA-approved vaccine or therapy that is available for the prevention and treatment. Traditional Chinese medicines (TCMs) represent a huge reservoir of bioactive chemicals and many TCMs have been shown to have antiviral activities. 373 extracts from 128 TCMs were evaluated using a high throughput assay to screen for inhibitors of Ebola virus cell entry. Extract of Rhodiola rosea displayed specific and potent inhibition against cell entry of both Ebola virus and Marburg virus. In addition, twenty commercial compounds that were isolated from Rhodiola rosea were evaluated using the pseudotyped Ebola virus entry assay, and it was found that ellagic acid and gallic acid, which are two structurally related compounds, are the most effective ones. The activity of the extract and the two pure compounds were validated using infectious Ebola virus. The time-of-addition experiments suggest that, mechanistically, the Rhodiola rosea extract and the effective compounds act at an early step in the infection cycle following initial cell attachment, but prior to viral/cell membrane fusion. Our findings provide evidence that Rhodiola rosea has potent anti-filovirus properties that may be developed as a novel anti-Ebola treatment.

Identification of Ellagic Acid from Plant Rhodiola rosea L. as an Anti-Ebola Virus Entry Inhibitor

PubMed Central

Cui, Qinghua; Du, Ruikun; Anantpadma, Manu; Schafer, Adam; Hou, Lin; Tian, Jingzhen; Cheng, Han; Rong, Lijun

2018-01-01

The recent 2014–2016 West African Ebola virus epidemic underscores the need for the development of novel anti-Ebola therapeutics, due to the high mortality rates of Ebola virus infections and the lack of FDA-approved vaccine or therapy that is available for the prevention and treatment. Traditional Chinese medicines (TCMs) represent a huge reservoir of bioactive chemicals and many TCMs have been shown to have antiviral activities. 373 extracts from 128 TCMs were evaluated using a high throughput assay to screen for inhibitors of Ebola virus cell entry. Extract of Rhodiola rosea displayed specific and potent inhibition against cell entry of both Ebola virus and Marburg virus. In addition, twenty commercial compounds that were isolated from Rhodiola rosea were evaluated using the pseudotyped Ebola virus entry assay, and it was found that ellagic acid and gallic acid, which are two structurally related compounds, are the most effective ones. The activity of the extract and the two pure compounds were validated using infectious Ebola virus. The time-of-addition experiments suggest that, mechanistically, the Rhodiola rosea extract and the effective compounds act at an early step in the infection cycle following initial cell attachment, but prior to viral/cell membrane fusion. Our findings provide evidence that Rhodiola rosea has potent anti-filovirus properties that may be developed as a novel anti-Ebola treatment. PMID:29584652

In Their Own Voices: Breaking the Vicious Cycle of Addiction, Treatment and Criminal Justice Among People who Inject Drugs in Ukraine

PubMed Central

Mazhnaya, Alyona; Bojko, Martha J.; Marcus, Ruthanne; Filippovych, Sergii; Islam, Zahedsul; Dvoriak, Sergey; Altice, Frederick L.

2016-01-01

Aims To understand how perceived law enforcement policies and practices contribute to the low rates of utilization of opioid agonist therapies (OAT) among people who inject drugs (PWIDs) in Ukraine. Methods Qualitative data from 25 focus groups (FGs) with 199 opioid-dependent PWIDs in Ukraine examined domains related to lived or learned experiences with OAT, police, arrest, incarceration, and criminal activity were analyzed using grounded theory principles. Findings Most participants were male (66%), in their late 30s, and previously incarcerated (85%) mainly for drug-related activities. When imprisoned, PWIDs perceived themselves as being “addiction-free”. After prison-release, the confluence of police surveillance, societal stress contributed to participants' drug use relapse, perpetuating a cycle of searching for money and drugs, followed by re-arrest and re-incarceration. Fear of police and arrest both facilitated OAT entry and simultaneously contributed to avoiding OAT since system-level requirements identified OAT clients as targets for police harassment. OAT represents an evidence-based option to ‘break the cycle’, however, law enforcement practices still thwart OAT capacity to improve individual and public health. Conclusion In the absence of structural changes in law enforcement policies and practices in Ukraine, PWIDs will continue to avoid OAT and perpetuate the addiction cycle with high imprisonment rates. PMID:27458326

Ecdysone signaling induces two phases of cell cycle exit in Drosophila cells

PubMed Central

Guo, Yongfeng; Flegel, Kerry; Kumar, Jayashree; McKay, Daniel J.

2016-01-01

ABSTRACT During development, cell proliferation and differentiation must be tightly coordinated to ensure proper tissue morphogenesis. Because steroid hormones are central regulators of developmental timing, understanding the links between steroid hormone signaling and cell proliferation is crucial to understanding the molecular basis of morphogenesis. Here we examined the mechanism by which the steroid hormone ecdysone regulates the cell cycle in Drosophila. We find that a cell cycle arrest induced by ecdysone in Drosophila cell culture is analogous to a G2 cell cycle arrest observed in the early pupa wing. We show that in the wing, ecdysone signaling at the larva-to-puparium transition induces Broad which in turn represses the cdc25c phosphatase String. The repression of String generates a temporary G2 arrest that synchronizes the cell cycle in the wing epithelium during early pupa wing elongation and flattening. As ecdysone levels decline after the larva-to-puparium pulse during early metamorphosis, Broad expression plummets, allowing String to become re-activated, which promotes rapid G2/M progression and a subsequent synchronized final cell cycle in the wing. In this manner, pulses of ecdysone can both synchronize the final cell cycle and promote the coordinated acquisition of terminal differentiation characteristics in the wing. PMID:27737823

Long noncoding RNA PANDA and scaffold-attachment-factor SAFA control senescence entry and exit.

PubMed

Puvvula, Pavan Kumar; Desetty, Rohini Devi; Pineau, Pascal; Marchio, Agnés; Moon, Anne; Dejean, Anne; Bischof, Oliver

2014-11-19

Cellular senescence is a stable cell cycle arrest that limits the proliferation of pre-cancerous cells. Here we demonstrate that scaffold-attachment-factor A (SAFA) and the long noncoding RNA PANDA differentially interact with polycomb repressive complexes (PRC1 and PRC2) and the transcription factor NF-YA to either promote or suppress senescence. In proliferating cells, SAFA and PANDA recruit PRC complexes to repress the transcription of senescence-promoting genes. Conversely, the loss of SAFA-PANDA-PRC interactions allows expression of the senescence programme. Accordingly, we find that depleting either SAFA or PANDA in proliferating cells induces senescence. However, in senescent cells where PANDA sequesters transcription factor NF-YA and limits the expression of NF-YA-E2F-coregulated proliferation-promoting genes, PANDA depletion leads to an exit from senescence. Together, our results demonstrate that PANDA confines cells to their existing proliferative state and that modulating its level of expression can cause entry or exit from senescence.

Low Cost Entry, Descent, and Landing (EDL) Instrumentation for Planetary Missions

NASA Technical Reports Server (NTRS)

Hwang, H. H.; Munk, M. M.; Dillman, R. A.; Mahzari, M.; Swanson, G. T.; White, T. R.

2016-01-01

Missions that involve traversing through a planetary atmosphere are unique opportunities that require elements of entry, descent, and landing (EDL). Many aspects of the EDL sequence are qualified using analysis and simulation due to the inability to conduct appropriate ground tests, however validating flight data are often lacking, especially for missions not involving Earth re-entry. NASA has made strategic decisions to collect EDL flight data in order to improve future mission designs. For example, MEDLI1 and EFT-1 gathered hypersonic pressure and in-depth temperature data in the thermal protection system (TPS). However, the ability to collect EDL flight data from the smaller competed missions, such as Discovery and New Frontiers, has been limited in part due to the Principal Investigator-managed cost-caps (PIMCC). The recent NASA decision to consider EDL instrumentation earlier in the mission design cycle led to the inclusion of a requirement in the Discovery 2014 Announcement of Opportunity which requires all missions that involve EDL to include an Engineering Science Investigation (ESI).2 The ESI would involve sensors for aerothermal environment and TPS; atmosphere, aerodynamics, and flight dynamics; atmospheric decelerator; and/or vehicle structure.3 The ESI activity would be funded outside of the PIMCC.

Integrin αvβ3 promotes infection by Japanese encephalitis virus.

PubMed

Fan, Wenchun; Qian, Ping; Wang, Dandan; Zhi, Xianwei; Wei, Yanming; Chen, Huanchun; Li, Xiangmin

2017-04-01

Japanese encephalitis virus (JEV) is a mosquito-borne flavivirus that is one of the major causes of viral encephalitis diseases worldwide. The JEV envelope protein facilitates viral entry, and its domain III contains an Arg-Gly-Asp (RGD) motif, that may modulate JEV entry through the RGD-binding integrin. In this study, the roles of integrin αv and β3 on the infection of JEV were evaluated. Reduced expression of integrin αv/β3 by special shRNA confers 2 to 4-fold inhibition of JEV replication in BHK-21 cells. Meanwhile, antibodies specific for integrin αv/β3 displayed ~58% and ~33% inhibition of JEV infectivity and RGD-specific peptides produced ~36% of inhibition. Expression of E protein and JEV RNA loads were clearly increased in CHO cells transfected with cDNA encoding human integrin β3. Moreover, integrin αv mediates JEV infection in viral binding stage of life cycle. Therefore, our study suggested that integrin αv and β3 serve as a host factor associated with JEV entry into the target cells. Copyright © 2016 Elsevier Ltd. All rights reserved.

Inflatable re-entry shield ready for test in space

NASA Astrophysics Data System (ADS)

2000-02-01

The Russian spacecraft Mars'96 for instance, which was launched in November 1996 but failed to reach its nominal orbit, carried two modules designed to land on that planet's surface. For the last part of the mission, an Inflatable Re-Entry and Descent Technology (IRDT) had been deployed. The main components of this system were an aerobraking and thermally protective shell, a densely packed inflating material and a pressurisation system. This technology is now considered applicable to other re-entry scenarios such as payload recovery from the International Space Station, planetary landers for science missions and atmospheric research. A demonstration mission on 9/10 February 2000 will evaluate the performance of this new technology before it is offered to potential users. A Russian Soyuz/Fregat launcher, lifting off from the Kazakh steppe near Baikonur, will provide a low-cost flight opportunity for the test vehicle, which is equipped with the inflatable heat shield and a sensor package developed by DaimlerChrysler Aerospace (DASA). After four orbits around the Earth, the test vehicle will be powered by the launcher's upper stage to re-enter the atmosphere for a landing the next day about 1800 km north-west of the launch site. During the mission, a number of technical parameters such as pressure, temperature and deceleration will be monitored and the inflation of the re-entry/descent structure observed. "From this novel technology, we are expecting a major breakthrough, to make re-entry of small payloads more and more reliable, simpler and less costly than traditional systems", explains Dieter Kassing, ESA's IRDT project manager. One of the main instruments on board the test vehicle is a sensor device developed by the University of Stuttgart for the determination of oxygen partial pressure in low Earth orbit and during re-entry. The scientific/technical investigations will be led by Dr. Ulrich Schoettle (Stuttgart University). Lionel Marraffa (ESA) will lead the evaluation of the IRDT's aerothermodynamic behaviour. DASA was responsible for integration of the sensor package and is ESA's co-investigator for evaluation of the application aspects of this new technology. In addition to the sensor package, the mission will accommodate a collection of special stones to study the physical and chemical modifications in sedimentary rocks, i.e. simulated meteorites, during atmospheric infall. Co-investors of this experiment are Dr. André Brack (CNRS, Orleans) and Dr. Gero Kurat (Vienna University). This experiment is being co-sponsored by ESA. The Russian/European Starsem launch company and NPO Lavochkin, the Russian company that developed the original IRDT technology, will be responsible for launch, orbit control, re-entry and recovery of the sensor package under contract with the International Science & Technology Centre (Moscow). ESA, the European Commission and DASA are co-funding this contract, contributing $600K each.

Glucose capped silver nanoparticles induce cell cycle arrest in HeLa cells.

PubMed

Panzarini, Elisa; Mariano, Stefania; Vergallo, Cristian; Carata, Elisabetta; Fimia, Gian Maria; Mura, Francesco; Rossi, Marco; Vergaro, Viviana; Ciccarella, Giuseppe; Corazzari, Marco; Dini, Luciana

2017-06-01

This study aims to determine the interaction (uptake and biological effects on cell viability and cell cycle progression) of glucose capped silver nanoparticles (AgNPs-G) on human epithelioid cervix carcinoma (HeLa) cells, in relation to amount, 2×10 3 or 2×10 4 NPs/cell, and exposure time, up to 48h. The spherical and well dispersed AgNPs (30±5nm) were obtained by using glucose as reducing agent in a green synthesis method that ensures to stabilize AgNPs avoiding cytotoxic soluble silver ions Ag + release. HeLa cells take up abundantly and rapidly AgNPs-G resulting toxic to cells in amount and incubation time dependent manner. HeLa cells were arrested at S and G2/M phases of the cell cycle and subG1 population increased when incubated with 2×10 4 AgNPs-G/cell. Mitotic index decreased accordingly. The dissolution experiments demonstrated that the observed effects were due only to AgNPs-G since glucose capping prevents Ag + release. The AgNPs-G influence on HeLa cells viability and cell cycle progression suggest that AgNPs-G, alone or in combination with chemotherapeutics, may be exploited for the development of novel antiproliferative treatment in cancer therapy. However, the possible influence of the cell cycle on cellular uptake of AgNPs-G and the mechanism of AgNPs entry in cells need further investigation. Copyright © 2017 Elsevier B.V. All rights reserved.

Space Debris Alert System for Aviation

NASA Astrophysics Data System (ADS)

Sgobba, Tommaso

2013-09-01

Despite increasing efforts to accurately predict space debris re-entry, the exact time and location of re-entry is still very uncertain. Partially, this is due to a skipping effect uncontrolled spacecraft may experience as they enter the atmosphere at a shallow angle. Such effect difficult to model depends on atmospheric variations of density. When the bouncing off ends and atmospheric re-entry starts, the trajectory and the overall location of surviving fragments can be precisely predicted but the time to impact with ground, or to reach the airspace, becomes very short.Different is the case of a functional space system performing controlled re-entry. Suitable forecasts methods are available to clear air and maritime traffic from hazard areas (so-called traffic segregation).In US, following the Space Shuttle Columbia accident in 2003, a re-entry hazard areas location forecast system was putted in place for the specific case of major malfunction of a Reusable Launch Vehicles (RLV) at re-entry. The Shuttle Hazard Area to Aircraft Calculator (SHAAC) is a system based on ground equipment and software analyses and prediction tools, which require trained personnel and close coordination between the organization responsible for RLV operation (NASA for Shuttle) and the Federal Aviation Administration. The system very much relies on the operator's capability to determine that a major malfunction has occurred.This paper presents a US pending patent by the European Space Agency, which consists of a "smart fragment" using a GPS localizer together with pre- computed debris footprint area and direct broadcasting of such hazard areas.The risk for aviation from falling debris is very remote but catastrophic. Suspending flight over vast swath of airspace for every re-entering spacecraft or rocket upper stage, which is a weekly occurrence, would be extremely costly and disruptive.The Re-entry Direct Broadcasting Alert System (R- DBAS) is an original merging and evolution of the Re- entry Breakup Recorder (REBR) concept developed by The Aerospace Corporation, often called the black box of spacecraft, and of the Shuttle Hazard Area to Aircraft Calculator (SHAAC). Unlike the REBR, whichdownloads data via satellite link for later analysis, the R-DBAS is intended as a direct communication tool with the end user. As a spacecraft carrying R-DBAS re- enters into the atmosphere, it relays a message with the coordinates of the falling debris footprint area to anyone with a receiver and a display like laptop or iPad, warning them of the hazard.Much like the REBR, the R-DBAS is designed to release from its host vehicle when it experiences significant heat which melts the attachment point and closes the power circuit. Once activated, the R-DBAS determines its own location and computes the final coordinates of the preloaded debris footprint which is then broadcasted to anyone holding a receiver in the proximity of the hazard area.The R-DBAS is intended to provide precise information directly to the cockpit. An airplane would have about 5- 7 minutes to get out of the way. Being the hazard area 1,000-2000 km long but very narrow, 30 -70 km, an escape manoeuvre from the risky area can be readily performed or go on holding before crossing the hazard area.By equipping aircraft and other vulnerable systems with a simple receivers that can be attached to a common laptop, escape manoeuvres can be performed as in front of bad weather or shelter can be taken by people on ground.

Building Bridges to the Economic Mainstream for African American Male Ex-Offenders: A Preliminary Assessment of an Inmate Education Re-Entry Program.

ERIC Educational Resources Information Center

Johnson, James H., Jr.; Farrell, Walter C., Jr.; Braithwaite, Lawrence P.

This paper describes a state-funded inmate education and re-entry program that provides soft skills training for soon-to-be released offenders. The paper presents preliminary evidence regarding the impact of this training on 14 young male participants. Data came from information prepared by inmates throughout the training program and ethnographic…

8 CFR 1212.2 - Consent to reapply for admission after deportation, removal or departure at Government expense.

Code of Federal Regulations, 2011 CFR

2011-01-01

... proof that he or she has remained outside of the United States for the time period required for re-entry... eligible to re-enter the United States. Any alien who has been deported or removed from the United States... permission to reapply for entry in conjunction with his or her application for adjustment of status. This...

Recognition of Tacit Skills: Sustaining Learning Outcomes in Adult Learning and Work Re-Entry

ERIC Educational Resources Information Center

Evans, Karen; Kersh, Natasha; Kontiainen, Seppo

2004-01-01

This paper is based on the project "Recognition of Tacit Skills and Knowledge in Work Re-entry" carried out as a part of the ESRC-funded Research Network "Improving Incentives to Learning in the Workplace". The network aims to contribute to improved practice among a wide range of practitioners. The study has investigated the part played by tacit…

8 CFR 1212.2 - Consent to reapply for admission after deportation, removal or departure at Government expense.

Code of Federal Regulations, 2010 CFR

2010-01-01

... proof that he or she has remained outside of the United States for the time period required for re-entry... eligible to re-enter the United States. Any alien who has been deported or removed from the United States... permission to reapply for entry in conjunction with his or her application for adjustment of status. This...

8 CFR 1212.2 - Consent to reapply for admission after deportation, removal or departure at Government expense.

Code of Federal Regulations, 2014 CFR

2014-01-01

... proof that he or she has remained outside of the United States for the time period required for re-entry... eligible to re-enter the United States. Any alien who has been deported or removed from the United States... permission to reapply for entry in conjunction with his or her application for adjustment of status. This...

8 CFR 1212.2 - Consent to reapply for admission after deportation, removal or departure at Government expense.

Code of Federal Regulations, 2012 CFR

2012-01-01

... proof that he or she has remained outside of the United States for the time period required for re-entry... eligible to re-enter the United States. Any alien who has been deported or removed from the United States... permission to reapply for entry in conjunction with his or her application for adjustment of status. This...

8 CFR 1212.2 - Consent to reapply for admission after deportation, removal or departure at Government expense.

Code of Federal Regulations, 2013 CFR

2013-01-01

... proof that he or she has remained outside of the United States for the time period required for re-entry... eligible to re-enter the United States. Any alien who has been deported or removed from the United States... permission to reapply for entry in conjunction with his or her application for adjustment of status. This...

An Evaluation of Factors Influencing the Academic Self-concept, Self-esteem and Academic Stress for Direct and Re-entry Students in Higher Education.

ERIC Educational Resources Information Center

Michie, Frances; Glachan, Martin; Bray, Diane

2001-01-01

Examines the differences in the undergraduate student experience of direct and re-entry students focusing on undergraduate students (n=112). Used a questionnaire to gather data to investigate the impact of age, gender, past school experiences, and motivation for participating in higher education based on academic self-esteem, self-concept, and…

Hydrogeology of the Faultless site, Nye County, Nevada

DOE Office of Scientific and Technical Information (OSTI.GOV)

Thordarson, W.

The Faultless event was the detonation of an intermediate-yield nuclear device on January 19, 1968, at a depth of 975 m below the surface of Hot Creek Valley, Nevada. This report presents details of the hydrogeology of the rubble chimney and radiochemical monitoring in re-entry hole UC-1-P-2SR. The surface location of re-entry hole UC-1-P-2SR is about 91 m north of the emplacement hole, UC-1. Re-entry hole UC-1-P-2SR was drilled to a total depth of about 1097 m. The hole penetrated Quaternary and Tertiary valley-fill sediments above the rubble chimney, as well as Quaternary and Tertiary valley-fill and Tertiary tuffaceous sedimentsmore » within the chimney and rubble-filled cavity. Monitoring of the water level in re-entry hole UC-1-P-2SR indicated that, from 1970 to 1974, the water level was 695 m below land surface. During filling of the rubble chimney from 1974 to 1983, the water level rose slowly to a depth of 335.1 m. The 1983 level was about 167 m below the pre-event level that was about 168 m below land surface. Water with temperatures ranging from 37 to 61/sup 0/C occurred at the bottom of the re-entry hole at depths ranging from 728 to 801 m. A temperature of 100/sup 0/C at a depth of 820 m was projected from temperature logs. The hydraulic connection between the re-entry hole and the rubble chimney is considered poor to fair. Chemical analyses of water samples indicate that the water predominantly was a sodium bicarbonate type. Chemical and radiochemical analyses indicated that, although the constituents generally increased with increasing depth, three distinct water-quality zones have lasted for more than 16 years, even during the rising water level. The hot, radioactive water from the Faultless event apparently rose into the lower zone concomitant with the rising water level, as the rubble chimney was being filled. This general rise was interrupted by the apparently major dilution from colder water descending from the upper zone during 1975 and 1977.« less

Characterizing the Anti-HIV Activity of Papuamide A

PubMed Central

Andjelic, Cynthia D; Planelles, Vicente; Barrows, Louis R

2008-01-01

Papuamide A is representative of a class of marine derived cyclic depsipeptides, reported to have cytoprotective activity against HIV-1 in vitro. We show here that papuamide A acts as an entry inhibitor, preventing human immunodeficiency virus infection of host cells and that this inhibition is not specific to R5 or X4 tropic virus. This inhibition of viral entry was determined to not be due to papuamide A binding to CD4 or HIV gp120, the two proteins involved in the cell-virus recognition and binding. Furthermore, papuamide A was able to inhibit HIV pseudotype viruses expressing envelope glycoproteins from vesicular stomatitis virus or amphotropic murine leukemia virus indicating the mechanism of viral entry inhibition is not HIV-1 envelope glycoprotein specific. Time delayed addition studies with the pseudotyped viruses show that papuamide A inhibits viral infection only at the initial stage of the viral life cycle. Additionally, pretreatment studies revealed that the virus, and not the cell, is the target of papuamide A’s action. Together, these results suggest a direct virucidal mechanism of HIV-1 inhibition by papuamide A. We also demonstrate here that the other papuamides (B-D) are able to inhibit viral entry indicating that the free amino moiety of 2,3-diaminobutanoic acid residue is not required for the virucidal activity. PMID:19172193

Immunohistochemical estimation of cell cycle phase in laryngeal neoplasia

PubMed Central

Chatrath, P; Scott, I S; Morris, L S; Davies, R J; Bird, K; Vowler, S L; Coleman, N

2006-01-01

We previously developed an immunohistochemical method for estimating cell cycle state and phase in tissue samples, including biopsies that are too small for flow cytometry. We have used our technique to examine whether primary abnormalities of the cell cycle exist in laryngeal neoplasia. Antibodies against the markers of cell cycle entry, minichromosome maintenance protein-2 (Mcm-2) and Ki67, and putative markers of cell cycle phase, cyclin D1 (G1-phase), cyclin A (S-phase), cyclin B1 (G2-phase) and phosphohistone H3 (Mitosis) were applied to paraffin-embedded sections of normal larynx (n=8), laryngeal dysplasia (n=10) and laryngeal squamous cell carcinoma (n=10). Cells expressing each marker were determined as a percentage of total cells, termed the labelling index (LI), and as a percentage of Mcm-2-positive cells, termed the labelling fraction (LF). The frequency of coexpression of each putative phase marker was investigated by confocal microscopy. There was a correlation between Mcm-2 and Ki67 LIs (ρ=0.93) but Mcm-2 LIs were consistently higher. All cells expressing a phase marker coexpressed Mcm-2, whereas Ki67 was not expressed in a proportion of these cells. The putative phase markers showed little coexpression. Labelling index values increased on progression from normal larynx through laryngeal dysplasia to squamous cell carcinoma for Mcm-2 (P=0.001), Ki67 (P=0.0002), cyclin D1 (P=0.015), cyclin A (P=0.0001) and cyclin B1 (P=0.0004). There was no evidence of an increase in the LF for any phase marker. Immunohistochemistry can be used to estimate cell cycle state and phase in laryngeal biopsies. Our data argues against primary cell cycle phase abnormalities in laryngeal neoplasia. PMID:16832409

Ki-67 Contributes to Normal Cell Cycle Progression and Inactive X Heterochromatin in p21 Checkpoint-Proficient Human Cells

PubMed Central

Sun, Xiaoming; Bizhanova, Aizhan; Matheson, Timothy D.; Yu, Jun; Zhu, Lihua Julie

2017-01-01

ABSTRACT The Ki-67 protein is widely used as a tumor proliferation marker. However, whether Ki-67 affects cell cycle progression has been controversial. Here we demonstrate that depletion of Ki-67 in human hTERT-RPE1, WI-38, IMR90, and hTERT-BJ cell lines and primary fibroblast cells slowed entry into S phase and coordinately downregulated genes related to DNA replication. Some gene expression changes were partially relieved in Ki-67-depleted hTERT-RPE1 cells by codepletion of the Rb checkpoint protein, but more thorough suppression of the transcriptional and cell cycle defects was observed upon depletion of the cell cycle inhibitor p21. Notably, induction of p21 upon depletion of Ki-67 was a consistent hallmark of cell types in which transcription and cell cycle distribution were sensitive to Ki-67; these responses were absent in cells that did not induce p21. Furthermore, upon Ki-67 depletion, a subset of inactive X (Xi) chromosomes in female hTERT-RPE1 cells displayed several features of compromised heterochromatin maintenance, including decreased H3K27me3 and H4K20me1 labeling. These chromatin alterations were limited to Xi chromosomes localized away from the nuclear lamina and were not observed in checkpoint-deficient 293T cells. Altogether, our results indicate that Ki-67 integrates normal S-phase progression and Xi heterochromatin maintenance in p21 checkpoint-proficient human cells. PMID:28630280

Respiratory syncytial virus (RSV) entry is inhibited by serine protease inhibitor AEBSF when present during an early stage of infection.

PubMed

Van der Gucht, Winke; Leemans, Annelies; De Schryver, Marjorie; Heykers, Annick; Caljon, Guy; Maes, Louis; Cos, Paul; Delputte, Peter L

2017-08-17

Host proteases have been shown to play important roles in many viral activities such as entry, uncoating, viral protein production and disease induction. Therefore, these cellular proteases are putative targets for the development of antivirals that inhibit their activity. Host proteases have been described to play essential roles in Ebola, HCV, HIV and influenza, such that specific protease inhibitors are able to reduce infection. RSV utilizes a host protease in its replication cycle but its potential as antiviral target is unknown. Therefore, we evaluated the effect of protease inhibitors on RSV infection. To measure the sensitivity of RSV infection to protease inhibitors, cells were infected with RSV and incubated for 18 h in the presence or absence of the inhibitors. Cells were fixed, stained and studied using fluorescence microscopy. Several protease inhibitors, representing different classes of proteases (AEBSF, Pepstatin A, E-64, TPCK, PMSF and aprotinin), were tested for inhibitory effects on an RSV A2 infection of HEp-2 cells. Different treatment durations, ranging from 1 h prior to inoculation and continuing for 18 h during the assay, were evaluated. Of all the inhibitors tested, AEBSF and TPCK significantly decreased RSV infection. To ascertain that the observed effect of AEBSF was not a specific feature related to HEp-2 cells, A549 and BEAS-2B cells were also used. Similar to HEp-2, an almost complete block in the number of RSV infected cells after 18 h of incubation was observed and the effect was dose-dependent. To gain insight into the mechanism of this inhibition, AEBSF treatment was applied during different phases of an infection cycle (pre-, peri- and post-inoculation treatment). The results from these experiments indicate that AEBSF is mainly active during the early entry phase of RSV. The inhibitory effect was also observed with other RSV isolates A1998/3-2 and A2000/3-4, suggesting that this is a general feature of RSV. RSV infection can be inhibited by broad serine protease inhibitors, AEBSF and TPCK. We confirmed that AEBSF inhibition is independent of the cell line used or RSV strain. The time point at which treatment with the inhibitor was most potent, was found to coincide with the expected moment of entry of the virion with the host cell.

Replicative stress and alterations in cell cycle checkpoint controls following acetaminophen hepatotoxicity restrict liver regeneration.

PubMed

Viswanathan, Preeti; Sharma, Yogeshwar; Gupta, Priya; Gupta, Sanjeev

2018-03-05

Acetaminophen hepatotoxicity is a leading cause of hepatic failure with impairments in liver regeneration producing significant mortality. Multiple intracellular events, including oxidative stress, mitochondrial damage, inflammation, etc., signify acetaminophen toxicity, although how these may alter cell cycle controls has been unknown and was studied for its significance in liver regeneration. Assays were performed in HuH-7 human hepatocellular carcinoma cells, primary human hepatocytes and tissue samples from people with acetaminophen-induced acute liver failure. Cellular oxidative stress, DNA damage and cell proliferation events were investigated by mitochondrial membrane potential assays, flow cytometry, fluorescence staining, comet assays and spotted arrays for protein expression after acetaminophen exposures. In experimental groups with acetaminophen toxicity, impaired mitochondrial viability and substantial DNA damage were observed with rapid loss of cells in S and G2/M and cell cycle restrictions or even exit in the remainder. This resulted from altered expression of the DNA damage regulator, ATM and downstream transducers, which imposed G1/S checkpoint arrest, delayed entry into S and restricted G2 transit. Tissues from people with acute liver failure confirmed hepatic DNA damage and cell cycle-related lesions, including restrictions of hepatocytes in aneuploid states. Remarkably, treatment of cells with a cytoprotective cytokine reversed acetaminophen-induced restrictions to restore cycling. Cell cycle lesions following mitochondrial and DNA damage led to failure of hepatic regeneration in acetaminophen toxicity but their reversibility offers molecular targets for treating acute liver failure. © 2018 John Wiley & Sons Ltd.

EXPERT: An atmospheric re-entry test-bed

NASA Astrophysics Data System (ADS)

Massobrio, F.; Viotto, R.; Serpico, M.; Sansone, A.; Caporicci, M.; Muylaert, J.-M.

2007-06-01

In recognition of the importance of an independent European access to the International Space Station (ISS) and in preparation for the future needs of exploration missions, ESA is conducting parallel activities to generate flight data using atmospheric re-entry test-beds and to identify vehicle design solutions for human and cargo transportation vehicles serving the ISS and beyond. The EXPERT (European eXPErimental Re-entry Test-bed) vehicle represents the major on-going development in the first class of activities. Its results may also benefit in due time scientific missions to planets with an atmosphere and future reusable launcher programmes. The objective of EXPERT is to provide a test-bed for the validation of aerothermodynamics models, codes and ground test facilities in a representative flight environment, to improve the understanding of issues related to analysis, testing and extrapolation to flight. The vehicle will be launched on a sub-orbital trajectory using a Volna missile. The EXPERT concept is based on a symmetrical re-entry capsule whose shape is composed of simple geometrical elements. The suborbital trajectory will reach 120 km altitude and a re-entry velocity of 5 6km/s. The dimensions of the capsule are 1.6 m high and 1.3 m diameter; the overall mass is in the range of 250 350kg, depending upon the mission parameters and the payload/instrumentation complement. A consistent number of scientific experiments are foreseen on-board, from innovative air data system to shock wave/boundary layer interaction, from sharp hot structures characterisation to natural and induced regime transition. Currently the project is approaching completion of the phase B, with Alenia Spazio leading the industrial team and CIRA coordinating the scientific payload development under ESA contract.

Re-building Daniell Cell with a Li-ion exchange Film

PubMed Central

Dong, Xiaoli; Wang, Yonggang; Xia, Yongyao

2014-01-01

Daniell cell (i.e. Zn-Cu battery) is widely used in chemistry curricula to illustrate how batteries work, although it has been supplanted in the late 19th century by more modern battery designs because of Cu2+-crossover-induced self-discharge and un-rechargeable characteristic. Herein, it is re-built by using a ceramic Li-ion exchange film to separate Cu and Zn electrodes for preventing Cu2+-crossover between two electrodes. The re-built Zn-Cu battery can be cycled for 150 times without capacity attenuation and self-discharge, and displays a theoretical energy density of 68.3 Wh kg−1. It is more important that both electrodes of the battery are renewable, reusable, low toxicity and environmentally friendly. Owing to these advantages mentioned above, the re-built Daniell cell can be considered as a promising and green stationary power source for large-scale energy storage. PMID:25369833

Coordination of Satellite Cell Activation and Self-Renewal by Par-Complex-Dependent Asymmetric Activation of p38α/β MAPK

PubMed Central

Troy, Andrew; Cadwallader, Adam B.; Fedorov, Yuri; Tyner, Kristina; Tanaka, Kathleen Kelly; Olwin, Bradley B.

2014-01-01

SUMMARY In response to muscle injury, satellite cells activate the p38α/β MAPK pathway to exit quiescence, then proliferate, repair skeletal muscle, and self-renew, replenishing the quiescent satellite cell pool. Although satellite cells are capable of asymmetric division, the mechanisms regulating satellite cell self-renewal are not understood. We found that satellite cells, once activated, enter the cell cycle and a subset undergoes asymmetric division, renewing the satellite cell pool. Asymmetric localization of the Par complex activates p38α/β MAPK in only one daughter cell, inducing MyoD, which permits cell cycle entry and generates a proliferating myoblast. The absence of p38α/β MAPK signaling in the other daughter cell prevents MyoD induction, renewing the quiescent satellite cell. Thus, satellite cells employ a mechanism to generate distinct daughter cells, coupling the Par complex and p38α/β MAPK signaling to link the response to muscle injury with satellite cell self-renewal. PMID:23040480

75 FR 5579 - Privacy Act of 1974; System of Records

Federal Register 2010, 2011, 2012, 2013, 2014

2010-02-03

... with re-entry controlled by passwords. The DLA Enterprise Hotline Program Database is also password...: * * * * * System location: Delete entry and replace with ``Director, DLA Accountability Office (DA), Headquarters....'' * * * * * Retention and disposal: Delete entry and replace with ``Records are destroyed/deleted 10 years after...

Platelet lysate activates quiescent cell proliferation and reprogramming in human articular cartilage: Involvement of hypoxia inducible factor 1.

PubMed

Nguyen, Van Thi; Cancedda, Ranieri; Descalzi, Fiorella

2018-03-01

The idea of rescuing the body self-repair capability lost during evolution is progressively gaining ground in regenerative medicine. In particular, growth factors and bioactive molecules derived from activated platelets emerged as promising therapeutic agents acting as trigger for repair of tissue lesions and restoration of tissue functions. Aim of this study was to assess the potential of a platelet lysate (PL) for human articular cartilage repair considering its activity on progenitor cells and differentiated chondrocytes. PL induced the re-entry in the cell cycle of confluent, growth-arrested dedifferentiated/progenitor cartilage cells. In a cartilage permissive culture environment, differentiated cells also resumed proliferation after exposure to PL. These findings correlated with an up-regulation of the proliferation/survival pathways ERKs and Akt and with an induction of cyclin D1. In short- and long-term cultures of articular cartilage explants, we observed a release of proliferating chondroprogenitors able to differentiate and form an "in vitro" tissue with properties of healthy articular cartilage. Moreover, in cultured cartilage cells, PL induced a hypoxia-inducible factor (HIF-1) alpha increase, its nuclear relocation and the binding to HIF-1 responsive elements. These events were possibly related to the cell proliferation because the HIF-1 inhibitor acriflavine inhibited HIF-1 binding to HIF-1 responsive elements and cell proliferation. Our study demonstrates that PL induces quiescent cartilage cell activation and proliferation leading to new cartilage formation, identifies PL activated pathways playing a role in these processes, and provides a rationale to the application of PL for therapeutic treatment of damaged articular cartilage. Copyright © 2017 John Wiley & Sons, Ltd.

Ceramic Foams for TPS Applications

NASA Technical Reports Server (NTRS)

Stockpoole, Mairead

2003-01-01

Ceramic foams have potential in many areas of Thermal Protection Systems (TPS) including acreage and tile leading edges as well as being suitable as a repair approach for re-entry vehicles. NASA Ames is conducting ongoing research in developing lower-density foams from pre-ceramic polymer routes. One of the key factors to investigate, when developing new materials for re-entry applications, is their oxidation behavior in the appropriate re-entry environment which can be simulated using ground based arc jet (plasma jet) testing. Arc jet testing is required to provide the appropriate conditions (stagnation pressures, heat fluxes, enthalpies, heat loads and atmospheres) encountered during flight. This work looks at the response of ceramic foams (Si systems) exposed to simulated reentry environments and investigates the influence of microstructure and composition on the material? response. Other foam properties (mechanical and thermal) will also be presented.

SHEFEX - the vehicle and sub-systems for a hypersonic re-entry flight experiment

NASA Astrophysics Data System (ADS)

Turner, John; Hörschgen, Marcus; Turner, Peter; Ettl, Josef; Jung, Wolfgang; Stamminger, Andreas

2005-08-01

The purpose of the Sharp Edge Flight Experiment (SHEFEX) is to investigate the aerodynamic behaviour and thermal problems of an unconventional shape for re-entry vehicles, comprising multi-facetted surfaces with sharp edges. The main object of this experiment is the correlation of numerical analysis with real flight data in terms of the aerodynamic effects and structural concept for the thermal protection system (TPS). The Mobile Rocket Base of the German Aerospace Center (DLR) is responsible for the test flight of SHEFEX on a two stage unguided solid propellant sounding rocket which is required to provide a velocity of the order of March 7 for more than 30 seconds during atmospheric re-entry. This paper discusses the problems associated with the mission requirements and the solutions developed for the vehicle and sub-systems.

A yeast gene essential for regulation of spindle pole duplication.

PubMed Central

Baum, P; Yip, C; Goetsch, L; Byers, B

1988-01-01

In eucaryotic cells, duplication of spindle poles must be coordinated with other cell cycle functions. We report here the identification in Saccharomyces cerevisiae of a temperature-sensitive lethal mutation, esp1, that deregulates spindle pole duplication. Mutant cells transferred to the nonpermissive temperature became unable to continue DNA synthesis and cell division but displayed repeated duplication of their spindle pole bodies. Although entry into this state after transient challenge by the nonpermissive temperature was largely lethal, rare survivors were recovered and found to have become increased in ploidy. If the mutant cells were held in G0 or G1 during exposure to the elevated temperature, they remained viable and maintained normal numbers of spindle poles. These results suggest dual regulation of spindle pole duplication, including a mechanism that promotes duplication as cells enter the division cycle and a negative regulatory mechanism, controlled by ESP1, that limits duplication to a single occurrence in each cell division cycle. Tetrad analysis has revealed that ESP1 resides at a previously undescribed locus on the right arm of chromosome VII. Images PMID:3072479

Cyclin C influences the timing of mitosis in fission yeast

PubMed Central

Banyai, Gabor; Szilagyi, Zsolt; Baraznenok, Vera; Khorosjutina, Olga; Gustafsson, Claes M.

2017-01-01

The multiprotein Mediator complex is required for the regulated transcription of nearly all RNA polymerase II–dependent genes. Mediator contains the Cdk8 regulatory subcomplex, which directs periodic transcription and influences cell cycle progression in fission yeast. Here we investigate the role of CycC, the cognate cyclin partner of Cdk8, in cell cycle control. Previous reports suggested that CycC interacts with other cellular Cdks, but a fusion of CycC to Cdk8 reported here did not cause any obvious cell cycle phenotypes. We find that Cdk8 and CycC interactions are stabilized within the Mediator complex and the activity of Cdk8-CycC is regulated by other Mediator components. Analysis of a mutant yeast strain reveals that CycC, together with Cdk8, primarily affects M-phase progression but mutations that release Cdk8 from CycC control also affect timing of entry into S phase. PMID:28515143

Mesenchymal stem cells inhibit dendritic cell differentiation and function by preventing entry into the cell cycle.

PubMed

Ramasamy, Rajesh; Fazekasova, Henrietta; Lam, Eric W-F; Soeiro, Inês; Lombardi, Giovanna; Dazzi, Francesco

2007-01-15

Mesenchymal stem cells (MSCs) play a crucial role in hematopoietic development and have been shown to exert a powerful immunosuppressive effect. In this study, we investigated the effect of bone marrow MSC on the differentiation and function of peripheral blood monocytes into dendritic cells (DCs). Human MSCs, generated from normal bone marrow, were added to peripheral blood monocytes stimulated in vitro with granulocyte-macrophage colony stimulating factor and interleukin-4 to become DCs. Monocytes were then examined for the expression of markers characteristic of DCs and their ability to stimulate allogeneic T cells. In addition, the effect of MSCs on the cell cycle of monocyte-derived DCs and the expression of various cell cycle proteins were analyzed by cytometric analysis and Western blotting with specific antibodies. MSCs blocked the differentiation of monocytes into DCs and impaired their antigen-presenting ability. This resulted from a block of monocytes from entering the G1 phase of the cell cycle with a progressive number of cells accumulating in the G0 phase. Cyclin D2 was downregulated. However, differently from what was observed in T-cells stimulated in the presence of MSCs, the expression of p27 was found decreased, suggesting the involvement of similar but not identical pathways. We conclude that MSCs impair monocyte differentiation and function by interfering with the cell cycle. These findings imply that MSC-induced immunosuppression might be a side product of a more general antiproliferative effect.

An Investigation of the Re-Entry Adjustment of Indians Who Studied in the U.S.A. Occasional Papers in Intercultural Learning, Number 17.

ERIC Educational Resources Information Center

Hansel, Bettina

This study explored the readjustment experience of 49 Indians who came to the United States to study and then returned to their home country. Interviews revealed that most experienced some stress or difficulty after their re-entry, with problems ranging from initial anxiety about getting a job or shock at the crowded conditions, pollution, or the…

HTV-4 undocking

NASA Image and Video Library

2013-09-04

One of the Expedition 36 crew members aboard the International Space Station took this picture of the Japanese HTV-4 unmanned cargo spacecraft,backdropped against the Earth,following its unberthing and release from the orbital outpost. HTV-4,after backing away from the flying complex,headed for re-entry into Earth's atmosphere,burning upon re-entry. Per Twitter message: And, shortly after release of #HTV4, flying over Africa (The storm clouds were amazing).

Space Shuttle Projects

NASA Image and Video Library

1986-01-01

Columbia, which opened the era of the Space Transportation System with four orbital flight tests, is featured in re-entry in the emblem designed by the STS-61C crew representing the seven team members who manned the vehicle for its seventh STS mission. Gold lettering against black background honors the astronaut crewmembers on the delta pattern surrounding colorful re-entry shock waves, and the payload specialists are honored similarly below the sphere

Procedure Planning: Anatomical Determinants of Strategy

PubMed Central

Hanratty, Colm; Walsh, Simon

2014-01-01

In contemporary practice there are three main methods that can be employed when attempting to open a chronic total occlusion (CTO) of a coronary artery; antegrade or retrograde wire escalation, antegrade dissection re-entry and retrograde dissection re-entry. This editorial will attempt to clarify the anatomical features that can be identified to help when deciding which of these strategies to employ initially and help understand the reasons for this decision. PMID:24694102

SHEFEX II - Aerodynamic Re-Entry Controlled Sharp Edge Flight Experiment

NASA Astrophysics Data System (ADS)

Longo, J. M. A.; Turner, J.; Weihs, H.

2009-01-01

In this paper the basic goals and architecture of the SHEFEX II mission is presented. Also launched by a two staged sounding rocket system SHEFEX II is a consequent next step in technology test and demonstration. Considering all experience and collected flight data obtained during the SHEFEX I Mission, the test vehicle has been re-designed and extended by an active control system, which allows active aerodynamic control during the re-entry phase. Thus, ceramic based aerodynamic control elements like rudders, ailerons and flaps, mechanical actuators and an automatic electronic control unit has been implemented. Special focus is taken on improved GNC Elements. In addition, some other experiments including an actively cooled thermal protection element, advanced sensor equipment, high temperature antenna inserts etc. are part of the SHEFEX II experimental payload. A final 2 stage configuration has been selected considering Brazilian solid rocket boosters derived from the S 40 family. During the experiment phase a maximum entry velocity of Mach around 10 is expected for 50 seconds. Considering these flight conditions, the heat loads are not representative for a RLV re-entry, however, it allows to investigate the principal behaviour of such a facetted ceramic TPS, a sharp leading edge at the canards and fins and all associated gas flow effects and their structural response.

Insights into Host Cell Modulation and Induction of New Cells by the Corn Smut Ustilago maydis.

PubMed

Redkar, Amey; Matei, Alexandra; Doehlemann, Gunther

2017-01-01

Many filamentous fungal pathogens induce drastic modulation of host cells causing abnormal infectious structures such as galls, or tumors that arise as a result of re-programming in the original developmental cell fate of a colonized host cell. Developmental consequences occur predominantly with biotrophic phytopathogens. This suggests that these host structures result as an outcome of efficient defense suppression and intimate fungal-host interaction to suit the pathogen's needs for completion of its infection cycle. This mini-review mainly summarizes host cell re-programming that occurs in the Ustilago maydis - maize interaction, in which the pathogen deploys cell-type specific effector proteins with varying activities. The fungus senses the physiological status and identity of colonized host cells and re-directs the endogenous developmental program of its host. The disturbance of host cell physiology and cell fate leads to novel cell shapes, increased cell size, and/or the number of host cells. We particularly highlight the strategies of U. maydis to induce physiologically varied host organs to form the characteristic tumors in both vegetative and floral parts of maize.

Overexpression of high molecular weight FGF-2 forms inhibits glioma growth by acting on cell-cycle progression and protein translation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lemiere, Sylvie; University Bordeaux1, Talence, F-33405; Azar, Rania

2008-12-10

In order to clarify the role of HMW FGF-2 in glioma development and angiogenesis, we over-expressed different human FGF-2 isoforms in C6 rat glioma cell line using a tetracycline-regulated expression system. Phenotypic modifications were analyzed in vitro and compared to untransfected cells or to cells over-expressing 18 kDa FGF-2 or all FGF-2 isoforms. In particular, we demonstrate that HMW FGF-2 has unique features in inhibiting glioma cell proliferation. HMW FGF-2 expressing cells showed a cell-cycle arrest at the G2M, demonstrating a role of HMW FGF-2 in controlling the entry in mitosis. Moreover, hydroxyurea was ineffective in blocking cells at themore » G1S boundary when HMW FGF-2 was expressed. We also show that the HMW FGF-2 isoforms inhibit 4E-BP1 phosphorylation at critical sites restoring the translation inhibitory activity of 4E-BP1. In vivo, inhibition of tumor growth was observed when cells expressed HMW FGF-2. This indicates that HMW FGF-2 inhibits tumor growth in glioma cells by acting on cell-cycle progression and protein translation.« less

Satellite cell response to concurrent resistance exercise and high-intensity interval training in sedentary, overweight/obese, middle-aged individuals.

PubMed

Pugh, Jamie K; Faulkner, Steve H; Turner, Mark C; Nimmo, Myra A

2018-02-01

Sarcopenia can begin from the 4-5th decade of life and is exacerbated by obesity and inactivity. A combination of resistance exercise (RE) and endurance exercise is recommended to combat rising obesity and inactivity levels. However, work continues to elucidate whether interference in adaptive outcomes occur when RE and endurance exercise are performed concurrently. This study examined whether a single bout of concurrent RE and high-intensity interval training (HIIT) alters the satellite cell response following exercise compared to RE alone. Eight sedentary, overweight/obese, middle-aged individuals performed RE only (8 × 8 leg extensions at 70% 1RM), or RE + HIIT (10 × 1 min at 90% HR max on a cycle ergometer). Muscle biopsies were collected from the vastus lateralis before and 96 h after the RE component to determine muscle fiber type-specific total (Pax7 + cells) and active (MyoD + cells) satellite cell number using immunofluorescence microscopy. Type-I-specific Pax7 + (P = 0.001) cell number increased after both exercise trials. Type-I-specific MyoD + (P = 0.001) cell number increased after RE only. However, an elevated baseline value in RE + HIIT compared to RE (P = 0.046) was observed, with no differences between exercise trials at 96 h (P = 0.21). Type-II-specific Pax7 + and MyoD + cell number remained unchanged after both exercise trials (all P ≥ 0.13). Combining a HIIT session after a single bout of RE does not interfere with the increase in type-I-specific total, and possibly active, satellite cell number, compared to RE only. Concurrent RE + HIIT may offer a time-efficient way to maximise the physiological benefits from a single bout of exercise in sedentary, overweight/obese, middle-aged individuals.

A proteomic chronology of gene expression through the cell cycle in human myeloid leukemia cells.

PubMed

Ly, Tony; Ahmad, Yasmeen; Shlien, Adam; Soroka, Dominique; Mills, Allie; Emanuele, Michael J; Stratton, Michael R; Lamond, Angus I

2014-01-01

Technological advances have enabled the analysis of cellular protein and RNA levels with unprecedented depth and sensitivity, allowing for an unbiased re-evaluation of gene regulation during fundamental biological processes. Here, we have chronicled the dynamics of protein and mRNA expression levels across a minimally perturbed cell cycle in human myeloid leukemia cells using centrifugal elutriation combined with mass spectrometry-based proteomics and RNA-Seq, avoiding artificial synchronization procedures. We identify myeloid-specific gene expression and variations in protein abundance, isoform expression and phosphorylation at different cell cycle stages. We dissect the relationship between protein and mRNA levels for both bulk gene expression and for over ∼6000 genes individually across the cell cycle, revealing complex, gene-specific patterns. This data set, one of the deepest surveys to date of gene expression in human cells, is presented in an online, searchable database, the Encyclopedia of Proteome Dynamics (http://www.peptracker.com/epd/). DOI: http://dx.doi.org/10.7554/eLife.01630.001.

A proteomic chronology of gene expression through the cell cycle in human myeloid leukemia cells

PubMed Central

Ly, Tony; Ahmad, Yasmeen; Shlien, Adam; Soroka, Dominique; Mills, Allie; Emanuele, Michael J; Stratton, Michael R; Lamond, Angus I

2014-01-01

Technological advances have enabled the analysis of cellular protein and RNA levels with unprecedented depth and sensitivity, allowing for an unbiased re-evaluation of gene regulation during fundamental biological processes. Here, we have chronicled the dynamics of protein and mRNA expression levels across a minimally perturbed cell cycle in human myeloid leukemia cells using centrifugal elutriation combined with mass spectrometry-based proteomics and RNA-Seq, avoiding artificial synchronization procedures. We identify myeloid-specific gene expression and variations in protein abundance, isoform expression and phosphorylation at different cell cycle stages. We dissect the relationship between protein and mRNA levels for both bulk gene expression and for over ∼6000 genes individually across the cell cycle, revealing complex, gene-specific patterns. This data set, one of the deepest surveys to date of gene expression in human cells, is presented in an online, searchable database, the Encyclopedia of Proteome Dynamics (http://www.peptracker.com/epd/). DOI: http://dx.doi.org/10.7554/eLife.01630.001 PMID:24596151

Perpetuation of torsade de pointes in heterogeneous hearts: competing foci or re-entry?

PubMed

Vandersickel, Nele; de Boer, Teun P; Vos, Marc A; Panfilov, Alexander V

2016-12-01

The underlying mechanism of torsade de pointes (TdP) remains of debate: perpetuation may be due to (1) focal activity or (2) re-entrant activity. The onset of TdP correlates with action potential heterogeneities in different regions of the heart. We studied the mechanism of perpetuation of TdP in silico using a 2D model of human cardiac tissue and an anatomically accurate model of the ventricles of the human heart. We found that the mechanism of perpetuation TdP depends on the degree of heterogeneity. If the degree of heterogeneity is large, focal activity alone can sustain a TdP, otherwise re-entrant activity emerges. This result can help to understand the relationship between the mechanisms of TdP and tissue properties and may help in developing new drugs against it. Torsade de pointes (TdP) can be the consequence of cardiac remodelling, drug effects or a combination of both. The mechanism underlying TdP is unclear, and may involve triggered focal activity or re-entry. Recent work by our group has indicated that both cases may exist, i.e. TdPs induced in the chronic atrioventricular block (CAVB) dog model may have a focal origin or are due to re-entry. Also it was found that heterogeneities might play an important role. In the current study we have used computational modelling to further investigate the mechanisms involved in TdP initiation and perpetuation, especially in the CAVB dog model, by the addition of heterogeneities with reduced repolarization reserve in comparison with the surrounding tissue. For this, the TNNP computer model was used for computations. We demonstrated in 2D and 3D simulations that ECGs with the typical TdP morphology can be caused by both multiple competing foci and re-entry circuits as a result of introduction of heterogeneities, depending on whether the heterogeneities have a large or a smaller reduced repolarization reserve in comparison with the surrounding tissue. Large heterogeneities can produce ectopic TdP, while smaller heterogeneities will produce re-entry-type TdP. © 2016 The Authors. The Journal of Physiology © 2016 The Physiological Society.

Asymmetric Distribution of Primary Cilia Allocates Satellite Cells for Self-Renewal.

PubMed

Jaafar Marican, Nur Hayati; Cruz-Migoni, Sara B; Borycki, Anne-Gaëlle

2016-06-14

Regeneration of vertebrate skeletal muscles requires satellite cells, a population of stem cells that are quiescent in normal conditions and divide, differentiate, and self-renew upon activation triggered by exercise, injury, and degenerative diseases. Satellite cell self-renewal is essential for long-term tissue homeostasis, and previous work has identified a number of external cues that control this process. However, little is known of the possible intrinsic control mechanisms of satellite cell self-renewal. Here, we show that quiescent satellite cells harbor a primary cilium, which is rapidly disassembled upon entry into the cell cycle. Contrasting with a commonly accepted belief, cilia reassembly does not occur uniformly in cells exiting the cell cycle. We found that primary cilia reassemble preferentially in cells committed to self-renew, and disruption of cilia reassembly causes a specific deficit in self-renewing satellite cells. These observations indicate that primary cilia provide an intrinsic cue essential for satellite cell self-renewal. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

Ablation of multi-wavelet re-entry: general principles and in silico analyses.

PubMed

Spector, Peter S; Correa de Sa, Daniel D; Tischler, Ethan S; Thompson, Nathaniel C; Habel, Nicole; Stinnett-Donnelly, Justin; Benson, Bryce E; Bielau, Philipp; Bates, Jason H T

2012-11-01

Catheter ablation strategies for treatment of cardiac arrhythmias are quite successful when targeting spatially constrained substrates. Complex, dynamic, and spatially varying substrates, however, pose a significant challenge for ablation, which delivers spatially fixed lesions. We describe tissue excitation using concepts of surface topology which provides a framework for addressing this challenge. The aim of this study was to test the efficacy of mechanism-based ablation strategies in the setting of complex dynamic substrates. We used a computational model of propagation through electrically excitable tissue to test the effects of ablation on excitation patterns of progressively greater complexity, from fixed rotors to multi-wavelet re-entry. Our results indicate that (i) focal ablation at a spiral-wave core does not result in termination; (ii) termination requires linear lesions from the tissue edge to the spiral-wave core; (iii) meandering spiral-waves terminate upon collision with a boundary (linear lesion or tissue edge); (iv) the probability of terminating multi-wavelet re-entry is proportional to the ratio of total boundary length to tissue area; (v) the efficacy of linear lesions varies directly with the regional density of spiral-waves. We establish a theoretical framework for re-entrant arrhythmias that explains the requirements for their successful treatment. We demonstrate the inadequacy of focal ablation for spatially fixed spiral-waves. Mechanistically guided principles for ablating multi-wavelet re-entry are provided. The potential to capitalize upon regional heterogeneity of spiral-wave density for improved ablation efficacy is described.

Realization of Minimum and Maximum Gate Function in Ta2O5-based Memristive Devices

NASA Astrophysics Data System (ADS)

Breuer, Thomas; Nielen, Lutz; Roesgen, Bernd; Waser, Rainer; Rana, Vikas; Linn, Eike

2016-04-01

Redox-based resistive switching devices (ReRAM) are considered key enablers for future non-volatile memory and logic applications. Functionally enhanced ReRAM devices could enable new hardware concepts, e.g. logic-in-memory or neuromorphic applications. In this work, we demonstrate the implementation of ReRAM-based fuzzy logic gates using Ta2O5 devices to enable analogous Minimum and Maximum operations. The realized gates consist of two anti-serially connected ReRAM cells offering two inputs and one output. The cells offer an endurance up to 106 cycles. By means of exemplary input signals, each gate functionality is verified and signal constraints are highlighted. This realization could improve the efficiency of analogous processing tasks such as sorting networks in the future.

Analysis of HSV viral reactivation in explants of sensory neurons

PubMed Central

Turner, Anne-Marie W.; Kristie, Thomas M.

2014-01-01

As with all Herpesviruses, Herpes simplex virus (HSV) has both a lytic replication phase and a latency-reactivation cycle. During lytic replication, there is an ordered cascade of viral gene expression that leads to the synthesis of infectious viral progeny. In contrast, latency is characterized by the lack of significant lytic gene expression and the absence of infectious virus. Reactivation from latency is characterized by the re-entry of the virus into the lytic replication cycle and the production of recurrent disease. This unit describes the establishment of the mouse sensory neuron model of HSV-1 latency-reactivation as a useful in vivo system for the analysis of mechanisms involved in latency and reactivation. Assays including the determination of viral yields, immunohistochemical/immunofluorescent detection of viral antigens, and mRNA quantitation are used in experiments designed to investigate the network of cellular and viral proteins regulating HSV-1 lytic infection, latency, and reactivation. PMID:25367271

Estimating the Stoichiometry of HIV Neutralization

PubMed Central

Magnus, Carsten; Regoes, Roland R.

2010-01-01

HIV-1 virions infect target cells by first establishing contact between envelope glycoprotein trimers on the virion's surface and CD4 receptors on a target cell, recruiting co-receptors, fusing with the cell membrane and finally releasing the genetic material into the target cell. Specific experimental setups allow the study of the number of trimer-receptor-interactions needed for infection, i.e., the stoichiometry of entry and also the number of antibodies needed to prevent one trimer from engaging successfully in the entry process, i.e., the stoichiometry of (trimer) neutralization. Mathematical models are required to infer the stoichiometric parameters from these experimental data. Recently, we developed mathematical models for the estimations of the stoichiometry of entry [1]. In this article, we show how our models can be extended to investigate the stoichiometry of trimer neutralization. We study how various biological parameters affect the estimate of the stoichiometry of neutralization. We find that the distribution of trimer numbers—which is also an important determinant of the stoichiometry of entry—influences the estimated value of the stoichiometry of neutralization. In contrast, other parameters, which characterize the experimental system, diminish the information we can extract from the data about the stoichiometry of neutralization, and thus reduce our confidence in the estimate. We illustrate the use of our models by re-analyzing previously published data on the neutralization sensitivity [2], which contains measurements of neutralization sensitivity of viruses with different envelope proteins to antibodies with various specificities. Our mathematical framework represents the formal basis for the estimation of the stoichiometry of neutralization. Together with the stoichiometry of entry, the stoichiometry of trimer neutralization will allow one to calculate how many antibodies are required to neutralize a virion or even an entire population of virions. PMID:20333245

The mechanistic effects of the dioxonaphthoimidazolium analog YM155 in renal cell carcinoma cell cycling and apoptosis.

PubMed

Sim, Mei Yi; Go, Mei Lin; Yuen, John Shyi Peng

2018-06-15

To investigate the effect of dioxonaphthoimidazolium analog YM155 on cell cycle progression of the clear-cell variant of renal cell carcinoma (ccRCC). Cell cycle analysis was performed using bromodeoxyuridine (BrdU) and PI, apoptosis initiation was monitored using Annexin V and proteins expression was determined using western immunoblotting. Here, we showed that YM155 activated stress-related molecules (histone H2AX, checkpoint kinases Chk1 and Chk2, p53) that mediate DNA damage checkpoint responses. The coordinated activation of these effector molecules disrupts progression of the cell cycle at the S phase as deduced from BrdU pulsing experiments and the ensuing changes in the levels of proteins (cyclins, CDKs, CDK inhibitors, phosphatases) that control cell cycle progression. Notably, we found increases in cyclin E and Cdc2 which regulate transition of cells from G1 to S, even as losses were observed for other CDKs and their cyclin partners. Furthermore, by inducing a loss in total pRb possibly by promoting its degradation, YM155 promoted the E2F transcription of genes that regulate entry into the S phase. After 24 h, cell cycle arrest to repair YM155-inflicted DNA damage was overtaken by p53-mediated apoptosis. YM155 induced increases in pro-apoptotic proteins (Bax and Bad), diminished anti-apoptotic proteins (Mcl-1, Bcl-xl, XIAP, survivin) and initiated cleavage of apoptotic marker proteins caspase 3 and PARP. Taken together, the added insight provided on the cell cycle perturbative effects of YM155 may assist clinicians in framing rational choices for combining YM155 with other anti-cancer drugs or treatment modalities in ccRCC. Copyright © 2018 Elsevier Inc. All rights reserved.

Coronas-F Orbit Monitoring and Re-Entry Prediction

NASA Technical Reports Server (NTRS)

Ivanov, N. M.; Kolyuka, Yu. F.; Afanasieva, T. I.; Gridchina, T. A.

2007-01-01

Russian scientific satellite CORONAS-F was launched on July, 31, 2001. The object was inserted in near-circular orbit with the inclination 82.5deg and a mean altitude approx. 520 km. Due to the upper atmosphere drag CORONAS-F was permanently descended and as a result on December, 6, 2005 it has finished the earth-orbital flight, having lifetime in space approx. 4.5 years. The satellite structural features and its flight attitude control led to the significant variations of its ballistic coefficient during the flight. It was a cause of some specific difficulties in the fulfillment of the ballistic and navigation support of this space vehicle flight. Besides the main mission objective CORONAS-F also has been selected by the Inter-Agency Space Debris Coordination Committee (IADC) as a target object for the next regular international re-entry test campaign on a program of surveillance and re-entry prediction for the hazard space objects within their de-orbiting phases. Spacecraft (S/C) CORONAS-F kept its working state right up to the end of the flight - down to the atmosphere entry. This fact enabled to realization of the additional research experiments, concerning with an estimation of the atmospheric density within the low earth orbits (LEO) of the artificial satellites, and made possible to continue track the S/C during final phase of its flight by means of Russian regular command & tracking system, used for it control. Thus there appeared a unique possibility of using for tracking S/C at its de-orbiting phase not only passive radar facilities, belonging to the space surveillance systems and traditionally used for support of the IADC re-entry test campaigns, but also more precise active trajectory radio-tracking facilities from the ground control complex (GCC) applied for this object. Under the corresponding decision of the Russian side such capability of additional high-precise tracking control of the CORONAS-F flight in this period of time has been implemented. The organizing of the CORONAS-F ballistic and navigational support (BNS) and solving its main tasks (such as S/C orbit determination (OD) and its motion prediction and connected with them) both for regular mission stage and for additional flight program were realized by the group of specialists from the Mission Control Center (MCC). MCC was also assigned as a principal organization from the Russian side for participation in the 7th IADC re-entry test campaign on CORONAS-F. The CORONAS-F flight features and space environments circumstances during its flight as well as a methodology and technology of spacecraft ballistic and navigational support are given below. The BNS results for different phases of S/C flight, including the results of its re-entry predictions, obtained during the realization of the 7th IADC test campaign are submitted. The accuracy of space vehicle re-entry prediction and its dependence on various factors are analyzed in more details.

Blastocoele expansion degree predicts live birth after single blastocyst transfer for fresh and vitrified/warmed single blastocyst transfer cycles.

PubMed

Du, Qing-Yun; Wang, En-Yin; Huang, Yan; Guo, Xiao-Yi; Xiong, Yu-Jing; Yu, Yi-Ping; Yao, Gui-Dong; Shi, Sen-Lin; Sun, Ying-Pu

2016-04-01

To evaluate the independent effects of the degree of blastocoele expansion and re-expansion and the inner cell mass (ICM) and trophectoderm (TE) grades on predicting live birth after fresh and vitrified/warmed single blastocyst transfer. Retrospective study. Reproductive medical center. Women undergoing 844 fresh and 370 vitrified/warmed single blastocyst transfer cycles. None. Live-birth rate correlated with blastocyst morphology parameters by logistic regression analysis and Spearman correlations analysis. The degree of blastocoele expansion and re-expansion was the only blastocyst morphology parameter that exhibited a significant ability to predict live birth in both fresh and vitrified/warmed single blastocyst transfer cycles respectively by multivariate logistic regression and Spearman correlations analysis. Although the ICM grade was significantly related to live birth in fresh cycles according to the univariate model, its effect was not maintained in the multivariate logistic analysis. In vitrified/warmed cycles, neither ICM nor TE grade was correlated with live birth by logistic regression analysis. This study is the first to confirm that the degree of blastocoele expansion and re-expansion is a better predictor of live birth after both fresh and vitrified/warmed single blastocyst transfer cycles than ICM or TE grade. Copyright © 2016. Published by Elsevier Inc.

Structural mechanisms of DREAM complex assembly and regulation

PubMed Central

Guiley, Keelan Z.; Liban, Tyler J.; Felthousen, Jessica G.; Ramanan, Parameshwaran

2015-01-01

The DREAM complex represses cell cycle genes during quiescence through scaffolding MuvB proteins with E2F4/5 and the Rb tumor suppressor paralog p107 or p130. Upon cell cycle entry, MuvB dissociates from p107/p130 and recruits B-Myb and FoxM1 for up-regulating mitotic gene expression. To understand the biochemical mechanisms underpinning DREAM function and regulation, we investigated the structural basis for DREAM assembly. We identified a sequence in the MuvB component LIN52 that binds directly to the pocket domains of p107 and p130 when phosphorylated on the DYRK1A kinase site S28. A crystal structure of the LIN52–p107 complex reveals that LIN52 uses a suboptimal LxSxExL sequence together with the phosphate at nearby S28 to bind the LxCxE cleft of the pocket domain with high affinity. The structure explains the specificity for p107/p130 over Rb in the DREAM complex and how the complex is disrupted by viral oncoproteins. Based on insights from the structure, we addressed how DREAM is disassembled upon cell cycle entry. We found that p130 and B-Myb can both bind the core MuvB complex simultaneously but that cyclin-dependent kinase phosphorylation of p130 weakens its association. Together, our data inform a novel target interface for studying MuvB and p130 function and the design of inhibitors that prevent tumor escape in quiescence. PMID:25917549

Impact of Rhenium-188, Gemcitabine, and 5-Fluorouracil on Cholangiocellular Carcinoma Cells: An In Vitro Study

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wiesinger, Benjamin, E-mail: Benjamin.wiesinger@med.uni-tuebingen.de; Farkas, Emese; Kehlbach, Rainer

2009-07-15

The purpose of this study was to compare the beneficial effects of radioactive stents and radioactive stents plus additional chemotherapy in the palliative treatment of cholangiocellular carcinomas. Cholangiocellular carcinoma cells (TFK-1 cells) were treated either with 8 Gy (RTB group) or 16 Gy (RTA group) {sup 188}Re or with {sup 188}Re irradiation (8 Gy) combined with either gemcitabine (8 Gy/Gem) or 5-fluorouracil (8 Gy/5-FU) at a dosage of 20 {mu}g/ml medium for 4 days and subsequently compared with an untreated control group. Proliferation kinetics were assessed on days 4, 7, 11, 18, 25, and 32. Colony formation assays were performedmore » on days 7, 18, and 32 and cell cycle distribution was examined on days 4, 7, 11, 15, 25, and 39. Cell proliferation kinetics showed the lowest cell numbers in the 8 Gy/5-FU group (control, 15,390,000; RTA group, 8,394,000; RTB group, 5,609,000; 8 Gy/Gem group, 423,000; and 8 Gy/5-FU group, 297,667). In contrast, clonogenic activity on day 32 was lower in the 8 Gy/Gem group (control, 29.3 colonies; RTB group, 23.1 colonies; 8 Gy/5-FU group, 21.5 colonies; 8 Gy/Gem, 3.3 colonies; and even augmented in the RTA group, with 37.7 colonies). Cell cycle distribution showed similar curves for all groups on slightly different levels except for the 8 Gy/5-FU group, which showed a relatively augmented percentage of cells on day 7 in the G2 M cycle phase and on day 4 in the S phase. In conclusion, irradiation (8 Gy) with {sup 188}Re administered, e.g., via coated stents, combined with Gem could be a valid option for the treatment of CCCs.« less

The timing of pronuclear formation, DNA synthesis and cleavage in the human 1-cell embryo.

PubMed

Capmany, G; Taylor, A; Braude, P R; Bolton, V N

1996-05-01

The timing of pronuclear formation and breakdown, DNA synthesis and cleavage during the first cell cycle of human embryogenesis are described. Pronuclei formed between 3 and 10 h post-insemination (hpi; median 8 hpi). S-phase commenced between 8 and 14 hpi, and was completed between 10 and 18 hpi. M-phase was observed between 22 and 31 hpi (median duration 3 h), and cleavage to the 2-cell stage took place between 25 and 33 hpi. The timing of the same events was determined in 1-cell embryos derived from re-inseminated human oocytes that had failed to fertilize during therapeutic in-vitro fertilization (IVF). In these embryos, pronuclei formed between 3 and 8 h post-re-insemination (hpr-i), coinciding with the beginning of S-phase. While S-phase was completed as early as 10 hpr-i in some embryos, it extended until at least 16 hpr-i in others. Pronuclear breakdown and cleavage occurred from 23 and 26 hpr-i respectively; however, they did not occur in some embryos until after 46 hpr-i. The results demonstrate a markedly greater degree of variation in the timing of these events in embryos derived from re-inseminated oocytes compared with embryos derived from conventional IVF, and thus throw into question the validity of using the former as models for studies of the first cell cycle of human embryogenesis.

Cell Cycle Status of CD34+ Hemopoietic Stem Cells Determines Lentiviral Integration in Actively Transcribed and Development-related Genes

PubMed Central

Papanikolaou, Eleni; Paruzynski, Anna; Kasampalidis, Ioannis; Deichmann, Annette; Stamateris, Evangelos; Schmidt, Manfred; von Kalle, Christof; Anagnou, Nicholas P

2015-01-01

Gene therapy utilizing lentiviral-vectors (LVs) is postulated as a dynamic therapeutic alternative for monogenic diseases. However, retroviral gene transfer may cause insertional mutagenesis. Although, such risks had been originally estimated as extremely low, several reports of leukemias or clonal dominance, have led to a re-evaluation of the mechanisms operating in insertional mutagenesis. Therefore, unraveling the mechanism of retroviral integration is mandatory toward safer gene therapy applications. In the present study, we undertook an experimental approach which enabled direct correlation of the cell cycle stage of the target cell with the integration profile of LVs. CD34+ cells arrested at different stages of cell cycle, were transduced with a GFP-LV. LAM-PCR was employed for integration site detection, followed by microarray analysis to correlate transcribed genes with integration sites. The results indicate that ~10% of integration events occurred in actively transcribed genes and that the cell cycle stage of target cells affects integration pattern. Specifically, use of thymine promoted a safer profile, since it significantly reduced integration within cell cycle-related genes, while we observed increased possibility for integration into genes related to development, and decreased possibility for integration within cell cycle and cancer-related genes, when transduction occurs during mitosis. PMID:25523760

ISG15 Functions as an Interferon-Mediated Antiviral Effector Early in the Murine Norovirus Life Cycle

PubMed Central

Rodriguez, Marisela R.; Monte, Kristen; Thackray, Larissa B.

2014-01-01

ABSTRACT Human noroviruses (HuNoV) are the leading cause of nonbacterial gastroenteritis worldwide. Similar to HuNoV, murine noroviruses (MNV) are enteric pathogens spread via the fecal-oral route and have been isolated from numerous mouse facilities worldwide. Type I and type II interferons (IFN) restrict MNV-1 replication; however, the antiviral effectors impacting MNV-1 downstream of IFN signaling are largely unknown. Studies using dendritic cells, macrophages, and mice deficient in free and conjugated forms of interferon-stimulated gene 15 (ISG15) revealed that ISG15 conjugation contributes to protection against MNV-1 both in vitro and in vivo. ISG15 inhibited a step early in the viral life cycle upstream of viral genome transcription. Directly transfecting MNV-1 RNA into IFN-stimulated mouse embryonic fibroblasts (MEFs) and bone marrow-derived dendritic cells (BMDC) lacking ISG15 conjugates bypassed the antiviral activity of ISG15, further suggesting that ISG15 conjugates restrict the MNV-1 life cycle at the viral entry/uncoating step. These results identify ISG15 as the first type I IFN effector regulating MNV-1 infection both in vitro and in vivo and for the first time implicate the ISG15 pathway in the regulation of early stages of MNV-1 replication. IMPORTANCE Type I IFNs are important in controlling murine norovirus 1 (MNV-1) infections; however, the proteins induced by IFNs that restrict viral growth are largely unknown. This report reveals that interferon-stimulated gene 15 (ISG15) mitigates MNV-1 replication both in vitro and in vivo. In addition, it shows that ISG15 inhibits MNV-1 replication by targeting an early step in the viral life cycle, MNV-1 entry and/or uncoating. These results identify ISG15 as the first type I IFN effector regulating MNV-1 infection both in vitro and in vivo and for the first time implicate the ISG15 pathway in the regulation of viral entry/uncoating. PMID:24899198

Cyclin D2 induces proliferation of cardiac myocytes and represses hypertrophy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Busk, Peter K.; Hinrichsen, Rebecca; Bartkova, Jirina

2005-03-10

The myocytes of the adult mammalian heart are considered unable to divide. Instead, mitogens induce cardiomyocyte hypertrophy. We have investigated the effect of adenoviral overexpression of cyclin D2 on myocyte proliferation and morphology. Cardiomyocytes in culture were identified by established markers. Cyclin D2 induced DNA synthesis and proliferation of cardiomyocytes and impaired hypertrophy induced by angiotensin II and serum. At the molecular level, cyclin D2 activated CDK4/6 and lead to pRB phosphorylation and downregulation of the cell cycle inhibitors p21{sup Waf1/Cip1} and p27{sup Kip1}. Expression of the CDK4/6 inhibitor p16 inhibited proliferation and cyclin D2 overexpressing myocytes became hypertrophic undermore » such conditions. Inhibition of hypertrophy by cyclin D2 correlated with downregulation of p27{sup Kip1}. These data show that hypertrophy and proliferation are highly related processes and suggest that cardiomyocyte hypertrophy is due to low amounts of cell cycle activators unable to overcome the block imposed by cell cycle inhibitors. Cell cycle entry upon hypertrophy may be converted to cell division by increased expression of activators such as cyclin D2.« less

Development of a Parachute System for Deceleration of Flying Vehicles in Supersonic Regimes

NASA Astrophysics Data System (ADS)

Pilyugin, N. N.; Khlebnikov, V. S.

2010-09-01

Aerodynamic problems arising during design and development of braking systems for re-entry vehicles are analyzed. Aerodynamic phenomena and laws valid in a supersonic flow around a pair of bodies having different shapes are studied. Results of this research can be used in solving application problems (arrangement and optimization of experiments; design and development of various braking systems for re-entry vehicles moving with supersonic speeds in the atmosphere).

Intermediate experimental vehicle, ESA program aerodynamics-aerothermodynamics key technologies for spacecraft design and successful flight

NASA Astrophysics Data System (ADS)

Dutheil, Sylvain; Pibarot, Julien; Tran, Dac; Vallee, Jean-Jacques; Tribot, Jean-Pierre

2016-07-01

With the aim of placing Europe among the world's space players in the strategic area of atmospheric re-entry, several studies on experimental vehicle concepts and improvements of critical re-entry technologies have paved the way for the flight of an experimental space craft. The successful flight of the Intermediate eXperimental Vehicle (IXV), under ESA's Future Launchers Preparatory Programme (FLPP), is definitively a significant step forward from the Atmospheric Reentry Demonstrator flight (1998), establishing Europe as a key player in this field. The IXV project objectives were the design, development, manufacture and ground and flight verification of an autonomous European lifting and aerodynamically controlled reentry system, which is highly flexible and maneuverable. The paper presents, the role of aerodynamics aerothermodynamics as part of the key technologies for designing an atmospheric re-entry spacecraft and securing a successful flight.

Incarcerated Veterans Outreach Program.

PubMed

Schaffer, Bradley J

2016-01-01

The objective of this study is to identify and facilitate re-entry services for military veterans in the Criminal Justice System through the Incarcerated Veteran Outreach Program. Veterans are explored as a subgroup of the general inmate jail populations in southern Ohio based upon veteran's status, military discharges, service-related injuries, treatment needs, pre-release planning, and re-entry services. Veterans reported having psycho-social problems, diverse levels of criminality, criminogenic needs, and significant episodes of homelessness. A sample of 399 incarcerated veterans in state prison, county jails, and community corrections setting were identified and completed the psycho-social pre-release assessment. Their average age was 44.6; they were more likely to be White males, divorced, most honorably discharged, and were represented in the following eras: 34% Vietnam, 35% post-Vietnam, 26% Persian Gulf War, and 5% Operation Iraqi Freedom/Operation Enduring Freedom. The findings encourage the development of a re-entry outreach model and strategies to prevent episodes of criminal recidivism.

Romance, recovery & community re-entry for criminal justice involved women: Conceptualizing and measuring intimate relationship factors and power

PubMed Central

Walt, Lisa C.; Hunter, Bronwyn; Salina, Doreen; Jason, Leonard

2015-01-01

Researchers have suggested that interpersonal relationships, particularly romantic relationships, may influence women’s attempts at substance abuse recovery and community re-entry after criminal justice system involvement. The present paper evaluates relational and power theories to conceptualize the influence of romantic partner and romantic relationship qualities on pathways in and out of substance abuse and crime. The paper then combines these conceptualizations with a complementary empirical analysis to describe an ongoing research project that longitudinally investigates these relational and power driven factors on women’s substance abuse recovery and community re-entry success among former substance abusing, recently criminally involved women. This paper is designed to encourage the integration of theory and empirical analysis by detailing how each of these concepts are operationalized and measured. Future research and clinical implications are also discussed. PMID:25750487

Thermal Analysis and Design of Multi-layer Insulation for Re-entry Aerodynamic Heating

NASA Technical Reports Server (NTRS)

Daryabeigi, Kamran

2001-01-01

The combined radiation/conduction heat transfer in high-temperature multi-layer insulations was modeled using a finite volume numerical model. The numerical model was validated by comparison with steady-state effective thermal conductivity measurements, and by transient thermal tests simulating re-entry aerodynamic heating conditions. A design of experiments technique was used to investigate optimum design of multi-layer insulations for re-entry aerodynamic heating. It was found that use of 2 mm foil spacing and locating the foils near the hot boundary with the top foil 2 mm away from the hot boundary resulted in the most effective insulation design. A 76.2 mm thick multi-layer insulation using 1, 4, or 16 foils resulted in 2.9, 7.2, or 22.2 percent mass per unit area savings compared to a fibrous insulation sample at the same thickness, respectively.

THE LARGE HIGH PRESSURE ARC PLASMA GENERATOR: A FACILITY FOR SIMULATING MISSLE AND SATELLITE RE-ENTRY. Research Report 56

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rose, P.; Powers, W.; Hritzay, D.

1959-06-01

The development of an arc wind tunnel capable of stagnation pressures in the excess of twenty atmospheres and using as much as fifteen megawatts of electrical power is described. The calibration of this facility shows that it is capable of reproducing the aerodynamic environment encountered by vehicles flying at velocities as great as satellite velocity. Its use as a missile re-entry material test facility is described. The large power capacity of this facility allows one to make material tests on specimens of size sufficient to be useful for material development yet at realistic energy and Reynolds number values. By themore » addition of a high-capacity vacuum system, this facility can be used to produce the low density, high Mach number environment needed for simulating satellite re-entry, as well as hypersonic flight at extreme altitudes. (auth)« less

On-ground casualty risk reduction by structural design for demise

NASA Astrophysics Data System (ADS)

Lemmens, Stijn; Funke, Quirin; Krag, Holger

2015-06-01

In recent years, awareness concerning the on-ground risk posed by un-controlled re-entering space systems has increased. On average over the past decade, an object with mass above 800 kg re-enters every week from which only a few, e.g. ESA's GOCE in 2013 and NASA's UARS in 2011, appeared prominent in international media. Space agencies and nations have discussed requirements to limit the on-ground risk for future missions. To meet the requirements, the amount of debris falling back on Earth has to be limited in number, mass and size. Design for demise (D4D) refers to all measures taken in the design of a space object to increase the potential for demise of the object and its components during re-entry. SCARAB (Spacecraft Atmospheric Re-entry and Break-Up) is ESA's high-fidelity tool which analyses the thermal and structural effects of atmospheric re-entry on spacecraft with a finite-element approach. For this study, a model of a representative satellite is developed in SCARAB to serve as test-bed for D4D analyses on a structural level. The model is used as starting point for different D4D approaches based on increasing the exposure of the satellite components to the aero-thermal environment, as a way to speed up the demise. Statistical bootstrapping is applied to the resulting on-ground fragment lists in order to compare the different re-entry scenarios and to determine the uncertainties of the results. Moreover, the bootstrap results can be used to analyse the casualty risk estimator from a theoretical point of view. The risk reductions for the analysed D4D techniques are presented with respect to the reference scenario for the modelled representative satellite.

Mimicking herpes simplex virus 1 and herpes simplex virus 2 mucosal behavior in a well-characterized human genital organ culture.

PubMed

Steukers, Lennert; Weyers, Steven; Yang, Xiaoyun; Vandekerckhove, Annelies P; Glorieux, Sarah; Cornelissen, Maria; Van den Broeck, Wim; Temmerman, Marleen; Nauwynck, Hans J

2014-07-15

We developed and morphologically characterized a human genital mucosa explant model (endocervix and ectocervix/vagina) to mimic genital herpes infections caused by herpes simplex virus types 1 (HSV-1) and 2 (HSV-2). Subsequent analysis of HSV entry receptor expression throughout the menstrual cycle in genital tissues was performed, and the evolution of HSV-1/-2 mucosal spread over time was assessed. Nectin-1 and -2 were expressed in all tissues during the entire menstrual cycle. Herpesvirus entry mediator expression was limited mainly to some connective tissue cells. Both HSV-1 and HSV-2 exhibited a plaque-wise mucosal spread across the basement membrane and induced prominent epithelial syncytia. © The Author 2014. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Identification of chemicals inducing cardiomyocyte proliferation in developmental stage-specific manner with pluripotent stem cells.

PubMed

Uosaki, Hideki; Magadum, Ajit; Seo, Kinya; Fukushima, Hiroyuki; Takeuchi, Ayako; Nakagawa, Yasuaki; Moyes, Kara White; Narazaki, Genta; Kuwahara, Koichiro; Laflamme, Michael; Matsuoka, Satoshi; Nakatsuji, Norio; Nakao, Kazuwa; Kwon, Chulan; Kass, David A; Engel, Felix B; Yamashita, Jun K

2013-12-01

The proliferation of cardiomyocytes is highly restricted after postnatal maturation, limiting heart regeneration. Elucidation of the regulatory machineries for the proliferation and growth arrest of cardiomyocytes is imperative. Chemical biology is efficient to dissect molecular mechanisms of various cellular events and often provides therapeutic potentials. We have been investigating cardiovascular differentiation with pluripotent stem cells. The combination of stem cell and chemical biology can provide novel approaches to investigate the molecular mechanisms and manipulation of cardiomyocyte proliferation. To identify chemicals that regulate cardiomyocyte proliferation, we performed a screening of a defined chemical library based on proliferation of mouse pluripotent stem cell-derived cardiomyocytes and identified 4 chemical compound groups: inhibitors of glycogen synthase kinase-3, p38 mitogen-activated protein kinase, and Ca(2+)/calmodulin-dependent protein kinase II, and activators of extracellular signal-regulated kinase. Several appropriate combinations of chemicals synergistically enhanced proliferation of cardiomyocytes derived from both mouse and human pluripotent stem cells, notably up to a 14-fold increase in mouse cardiomyocytes. We also examined the effects of identified chemicals on cardiomyocytes in various developmental stages and species. Whereas extracellular signal-regulated kinase activators and Ca(2+)/calmodulin-dependent protein kinase II inhibitors showed proliferative effects only on cardiomyocytes in early developmental stages, glycogen synthase kinase-3 and p38 mitogen-activated protein kinase inhibitors substantially and synergistically induced re-entry and progression of cell cycle in neonatal but also as well as adult cardiomyocytes. Our approach successfully uncovered novel molecular targets and mechanisms controlling cardiomyocyte proliferation in distinct developmental stages and offered pluripotent stem cell-derived cardiomyocytes as a potent tool to explore chemical-based cardiac regenerative strategies.

The Error-Prone DNA Polymerase κ Promotes Temozolomide Resistance in Glioblastoma through Rad17-Dependent Activation of ATR-Chk1 Signaling.

PubMed

Peng, Chenghao; Chen, Zhengxin; Wang, Shuai; Wang, Hong-Wei; Qiu, Wenjin; Zhao, Lin; Xu, Ran; Luo, Hui; Chen, Yuanyuan; Chen, Dan; You, Yongping; Liu, Ning; Wang, Huibo

2016-04-15

The acquisition of drug resistance is a persistent clinical problem limiting the successful treatment of human cancers, including glioblastoma (GBM). However, the molecular mechanisms by which initially chemoresponsive tumors develop therapeutic resistance remain poorly understood. In this study, we report that Pol κ, an error-prone polymerase that participates in translesion DNA synthesis, was significantly upregulated in GBM cell lines and tumor tissues following temozolomide treatment. Overexpression of Pol κ in temozolomide-sensitive GBM cells conferred resistance to temozolomide, whereas its inhibition markedly sensitized resistant cells to temozolomide in vitro and in orthotopic xenograft mouse models. Mechanistically, depletion of Pol κ disrupted homologous recombination (HR)-mediated repair and restart of stalled replication forks, impaired the activation of ATR-Chk1 signaling, and delayed cell-cycle re-entry and progression. Further investigation of the relationship between Pol κ and temozolomide revealed that Pol κ inactivation facilitated temozolomide-induced Rad17 ubiquitination and proteasomal degradation, subsequently silencing ATR-Chk1 signaling and leading to defective HR repair and the reversal of temozolomide resistance. Moreover, overexpression of Rad17 in Pol κ-depleted GBM cells restored HR efficiency, promoted the clearance of temozolomide-induced DNA breaks, and desensitized cells to the cytotoxic effects of temozolomide observed in the absence of Pol κ. Finally, we found that Pol κ overexpression correlated with poor prognosis in GBM patients undergoing temozolomide therapy. Collectively, our findings identify a potential mechanism by which GBM cells develop resistance to temozolomide and suggest that targeting the DNA damage tolerance pathway may be beneficial for overcoming resistance. Cancer Res; 76(8); 2340-53. ©2016 AACR. ©2016 American Association for Cancer Research.

Back home: a qualitative study exploring re-entering cross-cultural missionary aid workers' loss and grief.

PubMed

Selby, Susan; Moulding, Nicole; Clark, Sheila; Jones, Alison; Braunack-Mayer, Annette; Beilby, Justin

2009-01-01

Over 200 Australian, American, and British Non-Government Organizations send aid workers overseas including missionaries. On re-entry, they may suffer psychological distress; however, there is little research about their psychosocial issues and management in the family practice setting. Research suggests loss and grief as a suitable paradigm for family practitioners dealing with psychosocial issues. The aim of this study was to explore loss and grief issues for adult Australian missionary cross-cultural aid workers during their re-entry adjustment. Mixed methods were used and this study reports the qualitative method: semi-structured interviews conducted with 15 participants. Results were analyzed using framework analysis. Themes of re-entry loss and grief were identified with sub-themes of multiple varied losses, mechanisms of loss, loss of control, common grief phenomena, disenfranchised grief, and reactivation of past grief. Theoretical and clinical implications are discussed. Findings of this study suggest that loss and grief is an appropriate paradigm for the management of these workers in the family practice setting. Further research is needed to enable appropriate care.

Accelerated re-entrainment to advanced light cycles in BALB/cJ mice.

PubMed

Legates, Tara A; Dunn, Danielle; Weber, E Todd

2009-10-19

Circadian rhythms in mammals are coordinated by the suprachiasmatic nuclei (SCN) of the hypothalamus, which are most potently synchronized to environmental light-dark cycles. Large advances in the light-dark cycle typically yield gradual advances in activity rhythms on the order of 1-2h per day until re-entrainment is complete due to limitations on the circadian system which are not yet understood. In humans, this delay until re-entrainment is accomplished is experienced as jetlag, with accompanying symptoms of malaise, decreased cognitive performance, sleep problems and gastrointestinal distress. In these experiments, locomotor rhythms of BALB/cJ mice monitored by running wheels were shown to re-entrain to large 6- or 8-hour shifts of the light-dark cycle within 1-2 days, as opposed to the 5-7 days required for C57BL/6J mice. A single-day 6-hour advance of the LD cycle followed by release to constant darkness yielded similar phase shifts, demonstrating that exaggerated re-entrainment is not explained by masking of activity by the light-dark cycle. Responses in BALB/cJ mice were similar when monitored instead by motion detectors, indicating that wheel-running exercise does not influence the magnitude of responses. Neither brief (15 min) light exposure late during subjective nighttime nor 6-hour delays of the light-dark cycle produced exaggerated locomotor phase shifts, indicating that BALB/cJ mice do not merely experience enhanced sensitivity to light. Fos protein was expressed in cells of the SCN following acute light exposure at ZT10 of their previous light-dark cycle, a normally non-responsive time in the circadian cycle, but only in BALB/cJ (and not C57BL/6J) mice that had been subjected two days earlier to a single-day 6-hour advance of the light-dark cycle, indicating that their SCN had been advanced by that treatment. BALB/cJ mice may thus serve as a useful comparative model for studying molecular and physiological processes that limit responsiveness of circadian clocks to photic input.

An Automated Method to Compute Orbital Re-entry Trajectories with Heating Constraints

NASA Technical Reports Server (NTRS)

Zimmerman, Curtis; Dukeman, Greg; Hanson, John; Fogle, Frank R. (Technical Monitor)

2002-01-01

Determining how to properly manipulate the controls of a re-entering re-usable launch vehicle (RLV) so that it is able to safely return to Earth and land involves the solution of a two-point boundary value problem (TPBVP). This problem, which can be quite difficult, is traditionally solved on the ground prior to flight. If necessary, a nearly unlimited amount of time is available to find the 'best' solution using a variety of trajectory design and optimization tools. The role of entry guidance during flight is to follow the pre- determined reference solution while correcting for any errors encountered along the way. This guidance method is both highly reliable and very efficient in terms of onboard computer resources. There is a growing interest in a style of entry guidance that places the responsibility of solving the TPBVP in the actual entry guidance flight software. Here there is very limited computer time. The powerful, but finicky, mathematical tools used by trajectory designers on the ground cannot in general be converted to do the job. Non-convergence or slow convergence can result in disaster. The challenges of designing such an algorithm are numerous and difficult. Yet the payoff (in the form of decreased operational costs and increased safety) can be substantiaL This paper presents an algorithm that incorporates features of both types of guidance strategies. It takes an initial RLV orbital re-entry state and finds a trajectory that will safely transport the vehicle to Earth. During actual flight, the computed trajectory is used as the reference to be flown by a more traditional guidance method.

Overexpression of the growth arrest-specific homeobox gene Gax inhibits proliferation, migration, cell cycle progression, and apoptosis in serum-induced vascular smooth muscle cells.

PubMed

Zheng, H; Xue, S; Hu, Z L; Shan, J G; Yang, W G

2014-03-24

The Gax gene has been implicated in a variety of cell-developmental and biological processes, and aberrant Gax expression is linked to many diseases. In this study, to provide important insights for Gax-based gene therapy in vein graft restenosis and its anti-restenotic mechanism, we used rabbit vascular smooth muscle cells (VSMCs) to investigate the effects of Gax overexpression on proliferation, migration, cell cycle, and apoptosis in a serum-stimulated culture. Rabbit VSMC lines that stably overexpressed Gax were established by transfection with recombinant adenoviral vector Ad5-Gax. The effect of Gax overexpression on in vitro serum-induced VSMCs proliferation, migration, cell cycle, and apoptosis was assessed by MTT, wound healing, and flow cytometry assays, respectively. To investigate the effect of Gax overexpression on PCNA and MMP-2 in serum-induced VSMCs, immunocytochemistry, RT-PCR, and gelatin zymography were performed. The results clearly showed that Gax overexpression decreases PCNA expression in serum-induced VSMCs. Gax overexpression also significantly inhibited cell proliferation by blocking entry into the S-phase of the cell cycle, promoted cell apoptosis, and reduced cell migration activity by downregulating MMP-2 release and activity. These findings indicate that Gax would be an optimal target gene for gene therapy to treat vein graft restenosis.

Fiber Optic High Temperature Sensors for Re-Entry Vehicles

NASA Astrophysics Data System (ADS)

Haddad, E.; Kruzelecky, R.; Zou, J.; Wong, B.; Jamroz, W.; Sayeed, F.; Muylaert, J.-M.; McKenzie, I.

2009-01-01

MPB, within an ESA contract, is developing high temperature Fiber sensors (up to 1100°C) for re- ntry experiments, with direct application to the Thermo Protection Surface (TPS) of SHEFEX II. It addresses the challenges of obtaining high reflectivity FBG sensors, and integrating the fiber sensors within the selected TPS host material (C/SiC). Feasibility was demonstrated using free fiber sensors that showed the formation of the Chemical Composition Grating (CCG), with 80 % reflection at temperatures >750°C. The CCG grating was stable at high temperature (1000°C) for more than 50 hours, as well as after cycling between room temperature and 1000°C, with better than 0.5 % temperature accuracy (FBG central wavelength). Small FBG sensor packages were prepared and attached to C/SiC tiles. The calibration of the packaged fibers showed similar response to temperature as the free fiber sensor. The fiber sensor package was designed to maximize contact with the C/SiC surface to provide fast response to transients. Three- imension modeling with Ansys finite element analysis shows a time constant of 15-20 ms to reach 1200°C. A modular design will be implemented where a dedicated fiber line with 3 sensors and its own connector is used for each C/SiC tile. Small coupons of packaged sensors attached to C/SiC tiles will be tested in a re-entry environment at Von Karman Institute (Belgium) In a recently completed project with ESA, MPB developed and ground qualified a fiber sensor network, the "Fiber Sensor Demonstrator", that was successfully integrated as a payload with ESA's Proba-2. The system includes a central interrogation system that can be used to measure multiple parameters including a high temperature sensor for the Proba-2 thruster (up to 500°C).

Culture Medium Supplements Derived from Human Platelet and Plasma: Cell Commitment and Proliferation Support

PubMed Central

Muraglia, Anita; Nguyen, Van Thi; Nardini, Marta; Mogni, Massimo; Coviello, Domenico; Dozin, Beatrice; Strada, Paolo; Baldelli, Ilaria; Formica, Matteo; Cancedda, Ranieri; Mastrogiacomo, Maddalena

2017-01-01

Present cell culture medium supplements, in most cases based on animal sera, are not fully satisfactory especially for the in vitro expansion of cells intended for human cell therapy. This paper refers to (i) an heparin-free human platelet lysate (PL) devoid of serum or plasma components (v-PL) and (ii) an heparin-free human serum derived from plasma devoid of PL components (Pl-s) and to their use as single components or in combination in primary or cell line cultures. Human mesenchymal stem cells (MSC) primary cultures were obtained from adipose tissue, bone marrow, and umbilical cord. Human chondrocytes were obtained from articular cartilage biopsies. In general, MSC expanded in the presence of Pl-s alone showed a low or no proliferation in comparison to cells grown with the combination of Pl-s and v-PL. Confluent, growth-arrested cells, either human MSC or human articular chondrocytes, treated with v-PL resumed proliferation, whereas control cultures, not supplemented with v-PL, remained quiescent and did not proliferate. Interestingly, signal transduction pathways distinctive of proliferation were activated also in cells treated with v-PL in the absence of serum, when cell proliferation did not occur, indicating that v-PL could induce the cell re-entry in the cell cycle (cell commitment), but the presence of serum proteins was an absolute requirement for cell proliferation to happen. Indeed, Pl-s alone supported cell growth in constitutively activated cell lines (U-937, HeLa, HaCaT, and V-79) regardless of the co-presence of v-PL. Plasma- and plasma-derived serum were equally able to sustain cell proliferation although, for cells cultured in adhesion, the Pl-s was more efficient than the plasma from which it was derived. In conclusion, the cells expanded in the presence of the new additives maintained their differentiation potential and did not show alterations in their karyotype. PMID:29209609

Inhibition of Dengue Virus Entry into Target Cells Using Synthetic Antiviral Peptides

PubMed Central

Alhoot, Mohammed Abdelfatah; Rathinam, Alwin Kumar; Wang, Seok Mui; Manikam, Rishya; Sekaran, Shamala Devi

2013-01-01

Despite the importance of DENV as a human pathogen, there is no specific treatment or protective vaccine. Successful entry into the host cells is necessary for establishing the infection. Recently, the virus entry step has become an attractive therapeutic strategy because it represents a barrier to suppress the onset of the infection. Four putative antiviral peptides were designed to target domain III of DENV-2 E protein using BioMoDroid algorithm. Two peptides showed significant inhibition of DENV when simultaneously incubated as shown by plaque formation assay, RT-qPCR, and Western blot analysis. Both DET4 and DET2 showed significant inhibition of virus entry (84.6% and 40.6% respectively) using micromolar concentrations. Furthermore, the TEM images showed that the inhibitory peptides caused structural abnormalities and alteration of the arrangement of the viral E protein, which interferes with virus binding and entry. Inhibition of DENV entry during the initial stages of infection can potentially reduce the viremia in infected humans resulting in prevention of the progression of dengue fever to the severe life-threatening infection, reduce the infected vector numbers, and thus break the transmission cycle. Moreover these peptides though designed against the conserved region in DENV-2 would have the potential to be active against all the serotypes of dengue and might be considered as Hits to begin designing and developing of more potent analogous peptides that could constitute as promising therapeutic agents for attenuating dengue infection. PMID:23630436

Various supercritical carbon dioxide cycle layouts study for molten carbonate fuel cell application

NASA Astrophysics Data System (ADS)

Bae, Seong Jun; Ahn, Yoonhan; Lee, Jekyoung; Lee, Jeong Ik

2014-12-01

Various supercritical carbon dioxide (S-CO2) cycles for a power conversion system of a Molten Carbonate Fuel Cell (MCFC) hybrid system are studied in this paper. Re-Compressing Brayton (RCB) cycle, Simple Recuperated Brayton (SRB) cycle and Simple Recuperated Transcritical (SRT) cycle layouts were selected as candidates for this study. In addition, a novel concept of S-CO2 cycle which combines Brayton cycle and Rankine cycle is proposed and intensively studied with other S-CO2 layouts. A parametric study is performed to optimize the total system to be compact and to achieve wider operating range. Performances of each S-CO2 cycle are compared in terms of the thermal efficiency, net electricity of the MCFC hybrid system and approximate total volumes of each S-CO2 cycle. As a result, performance and total physical size of S-CO2 cycle can be better understood for MCFC S-CO2 hybrid system and especially, newly suggested S-CO2 cycle shows some success.

Targeting TRPM2 Channels Impairs Radiation-Induced Cell Cycle Arrest and Fosters Cell Death of T Cell Leukemia Cells in a Bcl-2-Dependent Manner

PubMed Central

Klumpp, Dominik; Misovic, Milan; Szteyn, Kalina; Shumilina, Ekaterina; Rudner, Justine; Huber, Stephan M.

2016-01-01

Messenger RNA data of lymphohematopoietic cancer lines suggest a correlation between expression of the cation channel TRPM2 and the antiapoptotic protein Bcl-2. The latter is overexpressed in various tumor entities and mediates therapy resistance. Here, we analyzed the crosstalk between Bcl-2 and TRPM2 channels in T cell leukemia cells during oxidative stress as conferred by ionizing radiation (IR). To this end, the effects of TRPM2 inhibition or knock-down on plasma membrane currents, Ca2+ signaling, mitochondrial superoxide anion formation, and cell cycle progression were compared between irradiated (0–10 Gy) Bcl-2-overexpressing and empty vector-transfected Jurkat cells. As a result, IR stimulated a TRPM2-mediated Ca2+-entry, which was higher in Bcl-2-overexpressing than in control cells and which contributed to IR-induced G2/M cell cycle arrest. TRPM2 inhibition induced a release from G2/M arrest resulting in cell death. Collectively, this data suggests a pivotal function of TRPM2 in the DNA damage response of T cell leukemia cells. Apoptosis-resistant Bcl-2-overexpressing cells even can afford higher TRPM2 activity without risking a hazardous Ca2+-overload-induced mitochondrial superoxide anion formation. PMID:26839633

8 CFR 1212.1 - Documentary requirements for nonimmigrants.

Code of Federal Regulations, 2011 CFR

2011-01-01

... valid re-entry permit issued by the Taiwan Ministry of Foreign Affairs. (2) Implementing regulations... satisfies the examining U.S. Immigration officer at the port-of-entry that he or she is clearly and beyond a... immigration officer's determination of admissibility at the port of entry at Guam; and (vi) Waives any right...

8 CFR 1212.1 - Documentary requirements for nonimmigrants.

Code of Federal Regulations, 2010 CFR

2010-01-01

... valid re-entry permit issued by the Taiwan Ministry of Foreign Affairs. (2) Implementing regulations... satisfies the examining U.S. Immigration officer at the port-of-entry that he or she is clearly and beyond a... immigration officer's determination of admissibility at the port of entry at Guam; and (vi) Waives any right...

9 CFR 93.215 - Special provisions.

Code of Federal Regulations, 2011 CFR

2011-01-01

..., That all poultry offered for re-entry upon examination by the veterinary inspector at the U.S. port of... be inspected at the border port of entry and, when accompanied by an import permit obtained under § 93.204 of this part and all conditions therein are observed, shall be allowed entry into the United...

9 CFR 93.215 - Special provisions.

Code of Federal Regulations, 2013 CFR

2013-01-01

..., That all poultry offered for re-entry upon examination by the veterinary inspector at the U.S. port of... be inspected at the border port of entry and, when accompanied by an import permit obtained under § 93.204 of this part and all conditions therein are observed, shall be allowed entry into the United...

8 CFR 1212.1 - Documentary requirements for nonimmigrants.

Code of Federal Regulations, 2014 CFR

2014-01-01

... valid re-entry permit issued by the Taiwan Ministry of Foreign Affairs. (2) Implementing regulations... satisfies the examining U.S. Immigration officer at the port-of-entry that he or she is clearly and beyond a... immigration officer's determination of admissibility at the port of entry at Guam; and (vi) Waives any right...

9 CFR 93.215 - Special provisions.

Code of Federal Regulations, 2014 CFR

2014-01-01

..., That all poultry offered for re-entry upon examination by the veterinary inspector at the U.S. port of... be inspected at the border port of entry and, when accompanied by an import permit obtained under § 93.204 of this part and all conditions therein are observed, shall be allowed entry into the United...

8 CFR 1212.1 - Documentary requirements for nonimmigrants.

Code of Federal Regulations, 2013 CFR

2013-01-01

... valid re-entry permit issued by the Taiwan Ministry of Foreign Affairs. (2) Implementing regulations... satisfies the examining U.S. Immigration officer at the port-of-entry that he or she is clearly and beyond a... immigration officer's determination of admissibility at the port of entry at Guam; and (vi) Waives any right...